• Title/Summary/Keyword: viability loss

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Neuroprotective Effects of Cheongnoemyeongsin-hwan against Hydrogen Peroxide-induced DNA Damage and Apoptosis in Human Neuronal-Derived SH-SY5Y Cells (인체 신경세포에서 청뇌명신환(淸腦明神丸)의 산화적 스트레스에 대한 세포보호 효과)

  • Pi, Guk Hyun;Hwang, Won Deuk
    • Herbal Formula Science
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    • v.25 no.1
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    • pp.51-68
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    • 2017
  • Objectives : Oxidative stress due to excessive accumulation of reactive oxygen species (ROS) is one of the risk factors for the development of several chronic diseases, including neurodegenerative diseases. Methods : In the present study, we investigated the protective effects of cheongnoemyeongsin-hwan (CNMSH) against oxidative stress‑induced cellular damage and elucidated the underlying mechanisms in neuronal-derived SH-SY5Y cells. Results : Our results revealed that treatment with CNMSH prior to hydrogen peroxide (H2O2) exposure significantly increased the SH-SY5Y cell viability, indicating that the exposure of the SH-SY5Y cells to CNMSH conferred a protective effect against oxidative stress. CNMSH also effectively attenuated H2O2‑induced comet tail formation, and decreased the phosphorylation levels of the histone ${\gamma}H2AX$, as well as the number of apoptotic bodies and Annexin V‑positive cells. In addition, CNMSH exhibited scavenging activity against intracellular ROS generation and restored the mitochondria membrane potential (MMP) loss that were induced by H2O2, suggesting that CNMSH prevents H2O2‑induced DNA damage and cell apoptosis. Moreover, H2O2 enhanced the cleavage of caspase-3 and degradation of poly (ADP-ribose)-polymerase, a typical substrate protein of activated caspase-3, as well as DNA fragmentation; however, these events were almost totally reversed by pretreatment with CNMSH. Furthermore, CNMSH increased the levels of heme oxygenase-1 (HO-1), which is a potent antioxidant enzyme, associated with the induction of nuclear factor-erythroid 2-related factor 2 (Nrf2). According to our data, CNMSH is able to protect SH-SY5Y cells from H2O2-induced apoptosis throughout blocking cellular damage related to oxidative stress through a mechanism that would affect ROS elimination and activating Nrf2/HO-1 signaling pathway. Conclusions : Therefore, we believed that CNMSH may potentially serve as an agent for the treatment and prevention of neurodegenerative diseases caused by oxidative stress.

Effect of Food Additives on Heat Sensitivity of Listeria monocytogenes H-12 and Decontamination of Kitchen Utensils (식품 첨가물이 Listeria monocytogenes H-12의 내열성에 미치는 영향 및 오염된 조리기구 제균)

  • LEE Hee Jung;LEE Tae Seek;SON Kwang Tae;BYUN Han Seok;KIM Ji Hoe;PARK Jeong Heum;PARK Mi Jung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.33 no.6
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    • pp.524-528
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    • 2000
  • Effect of food additives on the heat sensitivity of listeria monocytogenes H-12 inoculated into Pollack surimi was investigated and also confirmed the effectiveness of various decontamination method such as tap water washing, chlorination, ultraviolet irradiation and heat treatment haying been applied on cooking utensils. Food additives such as polyphosphate, chitosan, and potassium sorbate increased heat sensitivity of t monocytogenes H-12 and polyphosphate showed the strongest synergistic effect. The tested strain was not detected from stainless steel and plastic cutting board contaminated with $10^4{\~}10^5/cm^2$ of L monocytogenes H-12 after tap water washing for 10 seconds or 1 minute, but washing effect was not found in wooden cutting board. The chlorination of stainless steel and plastic cutting board for 10 seconds with $5{\~}50 ppm$ solution eliminated all cells of the contaminated strain, however any change of the viable cell count was not observed in the chlorination of wooden cutting board, W irradiation on stainless steel and plastic cutting board for 5 minutes with 15 W above 30 cm eliminated the contaminated strain, but the tested strain was still found after 60 seconds of irradiation on wooden cutting board. The treatment of hot water on all used cutting boards for 10 seconds at $70^{\circ}C$ resulted in complete loss of viability of the contaminated strain.

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Inhibition of PI3K/AKT Signaling Pathway Enhances Cordycepin-Induced Apoptosis in Human Gastric Cancer Cells (인체위암 세포에서 PI3K/AKT 신호 전달계 차단에 의한 동충하초 유래 Cordycepin의 Apoptosis 유발 효과 증진)

  • Lee, Hye Hyeon;Jeong, Jin-Woo;Choi, Yung Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.6
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    • pp.835-842
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    • 2016
  • The phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway plays a crucial role in cancer occurrence by promoting cell proliferation and inhibiting apoptosis. In the present study, we evaluated the effect of a PI3K inhibitor, LY294002, on the chemosensitivity of gastric cancer cells to cordycepin, a predominant functional component of the fungus Cordyceps militaris, in AGS human gastric cancer cells and investigated possible underlying cellular mechanisms. Our results revealed that cordycepin inhibited viability of AGS cells in a concentration-dependent manner and induced apoptosis, as determined by apoptotic cell morphologies and fluorescence-activated cell sorting analysis associated with attenuated activation of the PI3K/Akt signaling pathway. Treatment with cordycepin in combination with a subtoxic concentration of LY294002 enhanced cordycepin-induced cytotoxicity and apoptotic potentials in AGS cells. Sensitization of LY294002 to cordycepin-induced apoptosis was accompanied by activation of caspases (caspases-3, -8, and -9) and was concomitant with poly(ADP-ribose) polymerase cleavage. Moreover, LY294002 up-regulated pro-apoptotic Bax and enhanced truncation of Bid in cordycepin-treated AGS cells, which was connected with increased loss of mitochondrial membrane potential and release of cytochrome c from mitochondria to the cytosol. Taken together, these results indicate that inhibition of the PI3K/Akt signaling pathway could augment cordycepin-induced apoptosis in human gastric cancer cells by up-regulating caspase activity through mitochondrial dysfunction.

Effects of Newly Synthesized Recombinant Human Amyloid-β Complexes and Poly-Amyloid-β Fibers on Cell Apoptosis and Cognitive Decline

  • Park, Soojin;Huh, Jae-Won;Eom, Taekil;Park, Naeun;Lee, Youngjeon;Kim, Ju-Sung;Kim, Sun-Uk;Shim, Insop;Lee, Sang-Rae;Kim, Ekyune
    • Journal of Microbiology and Biotechnology
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    • v.27 no.11
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    • pp.2044-2051
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    • 2017
  • The main pathological hallmark of Alzheimer's disease is the deposition of amyloid-beta ($A{\beta}$) peptides in the brain. $A{\beta}$ has been widely used to mimic several aspects of Alzheimer's disease. However, several characteristics of amyloid-induced Alzheimer's disease pathology are not well established, especially in mice. The present study aimed to develop a new Alzheimer's disease model by investigating how $A{\beta}$ can be effectively aggregated using prokaryotes and eukaryotes. To express the $A{\beta}42$ complex in HEK293 cells, we cloned the $A{\beta}42$ region in a tandem repeat and incorporated the resulting construct into a eukaryotic expression vector. Following transfection into HEK293 cells via lipofection, cell viability assay and western blotting analysis revealed that exogenous $A{\beta}42$ can induce cell death and apoptosis. In addition, recombinant His-tagged $A{\beta}42$ was successfully expressed in Escherichia coli BL21 (DE3) and not only readily formed $A{\beta}$ complexes, but also inhibited the proliferation of SH-SY5Y cells and E. coli. For in vivo testing, recombinant His-tagged $A{\beta}42$ solution ($3{\mu}g/{\mu}l$ in $1{\times}PBS$ containing $1mM\;Ni^{2+}$) was injected stereotaxically into the left and right lateral ventricles of the brains of C57BL/6J mice (n = 8). Control mice were injected with $1{\times}PBS$ containing $1mM\;Ni^{2+}$ following the same procedure. Ten days after the sample injection, the Morris water maze test confirmed that exogenous $A{\beta}$ caused an increase in memory loss. These findings demonstrated that $Ni^{2+}$ is capable of complexing the 50-kDa amyloid and that intracerebroventricular injection of $A{\beta}42$ can lead to cognitive impairment, thereby providing improved Alzheimer's disease models.

Response of Germination Rate and Ascorbate Peroxidase Activity to Cryopreservation of Perilla (Perilla frutescens) Seeds with Variable Initial Viabilities (들깨 종자 활력 수준별 초저온 동결보존 후 발아율 및 Ascorbate Peroxidase 활성 변화)

  • Lee, Young-yi;Lee, Myeong-hee;Yi, Jung-yoon;Lee, Tae-yoon;Son, Eun-ho;Park, Hong-jae
    • Korean Journal of Environmental Agriculture
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    • v.36 no.4
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    • pp.256-262
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    • 2017
  • BACKGROUND: Seed of perilla (Perilla frutescens var. japonica Hara) is short-lived in conventional storage conditions. For long-term conservation of plant species, cryopreservation is the method currently available. This study was performed to find out reliable methods for a long-term storage of seeds of perilla as a genetic resource. METHODS AND RESULTS: Using seeds of 9 perilla cultivars, the effects of desiccation, aging, and cryopreservation on seed germinability and ascorbate peroxidase activity in the seeds were investigated. Initial germinability of the seeds was various, and dry seeds of all cultivars survived cryopreservation without loss of viability. The highest germination was achieved at 4-5% moisture content, and stimulatory effect of cryogenic temperature on the seed germination was observed in some cultivars. Accelerated aging of perilla seeds led to reduction in germination and ascorbate peroxidase activity, and the susceptibility of seeds to aging was different among the tested cultivars. No significant difference in germination was observed for the aged seeds of control and liquid nitrogen exposed. CONCLUSION: The results of this study suggest that cryopreservation at 4-5% moisture content would be a suitable method for long-term conservation of perilla seeds without detrimental effects on germination.

Antiviral Efficacy of Citra-kill®, Disinfectant Solution Against Avian Influenza Virus

  • Cha, Chun-Nam;Lee, Yeo-Eun;Kang, In-Jin;Yoo, Chang-Yeul;Park, Eun-Kee;An, Sun-Jeong;Kim, Suk;Lee, Hu-Jang
    • Journal of Food Hygiene and Safety
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    • v.27 no.1
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    • pp.18-23
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    • 2012
  • Highly pathogenic avian influenza virus (HPAIV) is already panzootic in poultry and caused a considerable economic loss in poultry industry. In addition, HPAIV continues to cross species barriers to infect humans and other mammals, often with fatal outcomes. In this study, the virucidal efficacy of Citra-$Kill^{(R)}$ composed to quaternary ammonium chloride and citric acid was investigated against avian influenza H9N2 virus (AIV). A virucidal efficacy was determined with the viability of AIV contacted with the disinfectant in the allantoic membrane of chicken embryos. Citra-$Kill^{(R)}$ and AIV was reacted on the distilled water (DW), hard water (HW) or organic matter suspension (OM) condition. On DW condition, AIV was inactivated with 2,000 fold dilutions of Citra-$Kill^{(R)}$. When the antiviral effect on HW condition was evaluated, the antiviral activity of the disinfectant showed on 1,500 fold dilutions against AIV. With the investigation of the antiviral effect of the disinfectant on OM condition, AIV was inactivated on 500 fold dilutions of Citra-$Kill^{(R)}$. As Citra-$Kill^{(R)}$ possesses virucidal efficacy against AIV, the disinfectant solution can be used to limit the spread of animal viral diseases.

The Conflict over the Separation of Prescribing and Dispensing Practice (SPDP) in Korea: A Bargaining Perspective (의약분업을 둘러싼 갈등 : 협상론의 관점에서)

  • Lee, Kyung-Won;Kim, Joung-Hwa;T. K. Ahn
    • Health Policy and Management
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    • v.12 no.4
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    • pp.91-113
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    • 2002
  • We report and analyze the Korean physicians' recent general strike over the implementation of the Separation of Prescribing and Dispensing Practice (SPDP) in which more than 18,000 private clinics and 280 hospitals participated. Utilizing game-theoretic models of bargaining we explain why the Korean physicians were so successful in organizing intense collective action against the government and securing very favorable policy outcomes. In particular, we highlight the role of distributional conflict among social actors in shaping the details of institutional reform. The introduction of the SPDP was a necessary first step in the overall reform of health care system in Korea. However, the SPDP was perceived to be a serious threat to the economic viability of their profession by the vast majority of Korean physicians who had long been relied on the profits from selling medicines to compensate for the loss of income due to the low service fee under the previous health care system. The strong political coalition among heterogeneous physicians enabled them to organize an intense form of collective action, the general strike. Thus, physicians were successful not only in dragging the government to a bargaining table, but also winning in the bargaining and securing an outcome vastly favorable to them. On the other hand, the lack of an overall reform plan in the health care policy area, especially the finance of the National Health Insurance and the need for maintaining an image as a successful reform initiator, motivated the government to reach a quick resolution with the striking physicians.

Anti-proliferation Effect of Coscinoderma sp. Extract on Human Colon Cancer Cells (Coscinoderma sp.의 대장암세포 증식 억제 효과)

  • Choi, Ki Heon;Jung, Joohee
    • Journal of Food Hygiene and Safety
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    • v.31 no.4
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    • pp.294-298
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    • 2016
  • Natural products are attractive as the source of new drug development. Especially, numerous unknown marine bioresources are an object of attention because the ocean occupies three fourth of the earth. Survival of marine bioresources in extreme environment may induce the production of biological active compounds. As previous study, we examined over 40 specimens of marine sponges collected from Micronesia and screened their anti-proliferative activities in various cancer cell lines. Among them, we investigated Coscinoderma sp.'s activity and mechanism in human colon carcinoma HCT116 and RKO cells. Furthermore, we also used the p53-knockout of HCT116 cells and the p53 loss of RKO cells for elucidating the role of p53. Coscinoderma sp. inhibited cellular viability independently of the p53 status. Therefore, we compared the expression level of cell death-related proteins by Coscinoderma sp. in HCT16 and in HCT116 p53KO cells. Coscinoderma sp. increased p53 level and NOXA levels and induced apoptosis under the condition of p53 existence. On the other hand, Coscinoderma sp. increased p21 and mTOR levels in HCT116 p53KO cells. These results suggest that Coscinoderma sp. induced anti-proliferation effect through different pathway depending on p53 status.

Reactive oxygen species-dependent apoptosis induction by water extract of Citrus unshiu peel in MDA-MB-231 human breast carcinoma cells

  • Kim, Min Yeong;Choi, Eun Ok;HwangBo, Hyun;Kwon, Da He;Ahn, Kyu Im;Kim, Hong Jae;Ji, Seon Yeong;Hong, Su-Hyun;Jeong, Jin-Woo;Kim, Gi Young;Park, Cheol;Choi, Yung Hyun
    • Nutrition Research and Practice
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    • v.12 no.2
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    • pp.129-134
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    • 2018
  • BACKGROUND/OBJECTIVES: Although several recent studies have reported the anti-cancer effects of extracts or components of Citrus unshiu peel, which has been used for various purposes in traditional medicine, the molecular mechanisms for their effects remain unclear. In the present study, the anti-cancer activity of a water-soluble extract of C. unshiu peel (WECU) in MDA-MB-231 human breast carcinoma cells at the level of apoptosis induction was investigated. MATERIALS/METHODS: Cytotoxicity was evaluated using the MTT assay. Apoptosis was detected using DAPI staining and flow cytometry analyses. Mitochondrial membrane potential, reactive oxygen species (ROS) assay, caspase activity and Western blotting were used to confirm the basis of apoptosis. RESULTS: The results indicated that WECU-induced apoptosis was related to the activation of caspase-8, and -9, representative initiator caspases of extrinsic and intrinsic apoptosis pathways, respectively, and caspase-3 accompanied by proteolytic degradation of poly(ADP-ribose) polymerase and down-regulation of the inhibitors of apoptosis protein family members. WECU also increased the pro-apoptotic BAX to anti-apoptotic BCL-2 ratio, loss of mitochondrial membrane potential and cytochrome c release from mitochondria to cytoplasm. Furthermore, WECU provoked the generation of ROS, but the reduction of cell viability and induction of apoptosis by WECU were prevented when ROS production was blocked by antioxidant N-acetyl cysteine. CONCLUSIONS: These results suggest that WECU suppressed proliferation of MDA-MB-231 cells by activating extrinsic and intrinsic apoptosis pathways in a ROS-dependent manner.

A Comparison between Water and Ethanol Extracts of Rumex acetosa for Protective Effects on Gastric Ulcers in Mice

  • Bae, Ji-Yeong;Lee, Yoon-Seok;Han, Sun-Young;Jeong, Eun-Ju;Lee, Mi-Kyeong;Kong, Jae-Yang;Lee, Dong-Hoon;Cho, Kyeong-Jae;Lee, Haeng-Soon;Ahn, Mi-Jeong
    • Biomolecules & Therapeutics
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    • v.20 no.4
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    • pp.425-430
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    • 2012
  • Rumex acetosa is a perennial herb that is widely distributed across eastern Asia. Although the hot water extract of R. acetosa has been used to treat gastritis or gastric ulcers as a folk medicine, no scientific report exists for the use of this plant to treat gastric ulcers. Hence, the present study was undertaken to assess the anti-ulcer activity of water and 70% ethanol extracts obtained from R. acetosa, using an HCl/ethanol-induced gastric ulcer model in mice. Anti-inflammatory and free radical-scavenging activities of these two extracts were also evaluated and compared. As a result, the administration of R. acetosa extracts significantly reduced the occurrence of gastric ulcers. However, significant differences in protective activity against gastric ulcers were observed between the two samples. In the case of the group pretreated with an ethanol extract dosage of 100 mg/kg, the protective effect (90.9%) was higher than that of water extract (41.2%). Under histological evaluation, pretreatment with R. acetosa extracts reversed negative effects, such as inflammation, edema, moderate hemorrhaging and loss of epithelial cells, presented by HCl/ethanol-treated stomachs. Meanwhile, R. acetosa extracts showed potent DPPH radical-scavenging activity and decreased NO production in a murine macrophage cell line, RAW 264.7, in a dose-dependent manner without affecting cellular viability. The greater anti-ulcer and NO production inhibitory activities exhibited by ethanol extracts compared to water extracts could be ascribed to the higher emodin levels, a major anthraquinone component of this plant.