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The Pro-apoptotic Effects of S100A8 and S100A9 in Human Monocytic Leukemia Cells, THP-1

  • Kim, In-Sik;Lee, Ji-Sook
    • Biomedical Science Letters
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    • v.24 no.2
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    • pp.134-137
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    • 2018
  • S100A8 and S100A9 are involved in pathogenesis of cancer by induction or inhibition of cancer as well as inflammation. In this study, we investigated the association of S100A8 and S100A9 with pathogenesis of leukemia using human monocytic leukemia cells, THP-1. The expression of TLR4, which is a known receptor of S100A8 and S100A9, was examined by using flow cytometry and Western blotting. THP-1 cells have high surface and cytosol expression of TLR4. S100A8 and S100A9 suppressed the cell survival, and this suppression was found to be associated with apoptosis because they increased the number of apoptotic cells in a dose- and a time-dependent manners. However, S100A8 and S100A9 had no effect on the survival and apoptosis of monocytes isolated from the peripheral blood. We next examined the apoptotic effect of lipopolysaccharide (LPS) and monophosphoryl lipid A (MPLA), which are other ligands of TLR4, in THP-1 cells. Lipopolysaccharide had no effect on cell survival, but MPLA is effective on the cell apoptosis. These results suggest that S100A8 and S100A9 may regulate leukemia cell survival via TLR4, which is an essential receptor in the pro-apoptotic mechanism induced by S100A8 and S100A9. These findings may shed light on development of a possible therapeutic drug for leukemia treatment.

Induction of Autophagy and Apoptosis by the Roots of Platycodon grandiflorum on NCI-H460 Human Non-small Lung Carcinoma Cells (길경(桔梗)에 의한 NCI-H460 인체 비소세포폐암 세포에서의 autophagy 및 apoptosis 유발 효과)

  • Hong, Su-Hyun;Han, Min-Ho;Park, Cheol;Park, Sang-Eun;Hong, Sang-Hoon;Choi, Yung-Hyun
    • The Journal of Internal Korean Medicine
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    • v.35 no.3
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    • pp.317-331
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    • 2014
  • Objectives: The root of Platycodon grandiflorum (PG) has been known to possess a range of pharmacological activities including anti-cancer, anti-inflammatory, and anti-oxidant effects. The present study was designed to investigate whether or not PG-induced cell death was connected with autophagy and apoptosis in NCI-H460 human lung cancer cells. Methods: Effects on the cell viability and apoptotic activity were quantified using MTT assays and flow cytometry analysis, respectively. Protein activation was measured by immunoblotting. Autophagy was measured by LC3 immunofluorescence and immunoblotting. ROS production and loss of mitochondria membrane potential (MMP) were checked with flow cytometry analysis. Results: Following exposure to PG, NCI-H460 cell proliferation decreased simultaneously inducing autophagic vacuoles and up-regulation of microtubule-associated protein 1 light chain 3 and beclin-1 protein expressions. Interestingly, pre-treated with autophagy inhibitors, 3-methyladenin or bafilomycin A1 further triggered reduction of cell viability. PG treatment also induced apoptosis that was related modulation of Bcl-2 family proteins, death receptors and activation of caspases. In addition, PG stimulation clearly enhanced loss of MMP and reactive oxygen species (ROS) generation. Conclusions: Our results suggest that PG elicited both autophagy and apoptosis by increasing loss of MMP and ROS production. PG induced-autophagy may play a cell protective role.

Combinatorial Antitumor Activity of Oxaliplatin with Epigenetic Modifying Agents, 5-Aza-CdR and FK228, in Human Gastric Cancer Cells

  • Park, Jong Kook;Seo, Jung Seon;Lee, Suk Kyeong;Chan, Kenneth K;Kuh, Hyo-Jeong
    • Biomolecules & Therapeutics
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    • v.26 no.6
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    • pp.591-598
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    • 2018
  • Epigenetic silencing is considered to be a major mechanism for loss of activity in tumor suppressors. Reversal of epigenetic silencing by using inhibitors of DNA methyltransferase (DNMT) or histone deacetylases (HDACs) such as 5-Aza-CdR and FK228 has shown to enhance cytotoxic activities of several anticancer agents. This study aims to assess the combinatorial effects of genesilencing reversal agents (5-Aza-CdR and FK228) and oxaliplatin in gastric cancer cells, i.e., Epstein-Barr virus (EBV)-negative SNU-638 and EBV-positive SNU-719 cells. The doublet combinatorial treatment of 5-Aza-CdR and FK228 exhibited synergistic effects in both cell lines, and this was further corroborated by Zta expression induction in SNU-719 cells. Three drug combinations as 5-Aza-CdR/FK228 followed by oxaliplatin, however, resulted in antagonistic effects in both cell lines. Simultaneous treatment with FK228 and oxaliplatin induced synergistic and additive effects in SNU-638 and SNU-719 cells, respectively. Three drug combinations as 5-Aza-CdR prior to FK228/oxaliplatin, however, again resulted in antagonistic effects in both cell lines. This work demonstrated that efficacy of doublet synergistic combination using DNMT or HDACs inhibitors can be compromised by adding the third drug in pre- or post-treatment approach in gastric cancer cells. This implies that the development of clinical trial protocols for triplet combinations using gene-silencing reversal agents should be carefully evaluated in light of their potential antagonistic effects.

Identification and Characterization of an Oil-degrading Yeast, Yarrowia lipolytica 180

  • Kim, Tae-Hyun;Lee, Jung-Hyun;Oh, Young-Sook;Bae, Kyung-Sook;Kim, Sang-Jin
    • Journal of Microbiology
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    • v.37 no.3
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    • pp.128-135
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    • 1999
  • Among oil-degrading microorganisms isolated from oil-polluted industrial areas, one yeast strain showed high degradation activity of aliphatic hydrocarbons. From the analyses of 18S rRNA sequences, fatty acid, coenzyme Q system, G+C content of DNA, and biochemical characteristics, the strain was identified as Yarrowia lipolytica 180. Y. lipolytica 180 degraded 94% of aliphatic hydrocarbons in minimal salts medium containing 0.2% (v/v) of Arabian light crude oil within 3 days at 25$^{\circ}C$. Optimal growth conditions for temperature, pH, NaCl concentration, and crude oil concentration were 30$^{\circ}C$, pH 5-7, 1%, and 2% (v/v), respectively. Y. lipolytica 180 reduced surface tension when cultured on hydrocarbon substrates (1%, v/v), and the measured values of the surface tension were in the range of 51 to 57 dynes/cm. Both the cell free culture broth and cell debris of Y. lipolytica 180 were capable of emulsifying 2% (v/v) crude oil by itself. They were also capable of degrading crude oil (2%). The strain showed a cell surface hydrophobicity higher than 90%, which did not require hydrocarbon substrates for its induction. These results suggest that Y. lipolytica has high oil-degrading activity through its high emulsifying activity and cell hydrophobicity, and further indicate that the cell surface is responsible for the metabolism of aliphatic hydrocarbons.

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The Inhibitory Effects of Trichosanthes kirilowii Root against Cadmium Induced Cytotoxicity (III) (천화분의 카드뮴에 대한 독성억제효과(III))

  • Lee, Jeong-Ho;You, Il-Soo;Kim, Shin-Kee;Lee, Ki-Nam;Chung, Woo-Young;Han, Du-Seok;Baek, Seung-Hwa
    • Korean Journal of Pharmacognosy
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    • v.32 no.1 s.124
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    • pp.15-21
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    • 2001
  • This study was conducted to investigate the antitoxic components in the water extract of the roots of Trichosanthes kirilowii (Cucurbitaceae). The results were as follows: Generally, detoxication effects by the water extract of T. kirilowii increased in proportion to the concentrations. Experimental animals were treated with cadmium and T. kirilowii water extract by oral administration. When 40 mg/kg dosage of T. kirilowii extract was administrated it showed the highest antitoxic effects in metallothionein induction. After the water extract treatment, body weights did not increase in proportion to the extract concentrations. These results suggest that T. kirilowii extract increased metallothionein concentration and decreased the toxicity of cadmium in rats. In vitro the antitoxic activity of water extract of T. kirilowii on NIH 3T3 fibroblasts was evaluated by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide) and SRB (sulforhodamine B protein) assays. The light microscopic study was carried out to observe morphological changes of the treated cells. These results were obtained as follows; The concentration of $10^{-2}\;mg/ml$ of T. kirilowii extract was shown significant antitoxic activity. The number of NIH 3T3 fibroblasts were increased and tend to regenerate. These results suggest that T. kirilowii extract retains a potential antitoxic activity.

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The Inhibitory Effects of the Methanol Extract of Houttuynia cordata $T_{HUNB}$ against Cadmium Induced Cytotoxicity (V) (어성초 메탄올 추출물의 카드뮴에 대한 독성억제효과(V))

  • Lee, Jeong-Ho;Jeong, Seung-Il;You, Il-Soo;Kim, Shin-Kee;Lee, Ki-Nam;Han, Du-Seok;Baek, Seung-Hwa
    • Korean Journal of Pharmacognosy
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    • v.32 no.1 s.124
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    • pp.61-67
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    • 2001
  • This study was conducted to investigate the antitoxic agent in methanol extract of Houttuynia cordata $T_{HUNB}$. Detoxication effects By H. cordata $T_{HUNB}$ extract increased in proportion to the extract concentrations. When 40 mg/kg dosage of H. cordata $T_{HUNB}$ extract was administered, it showed the highest antitoxic effects in metallothionein induction. After the extract treatment, body weights generally increased in proportion to the extract concentrations. From the above results, H. cordata $T_{HUNB}$ extract increased metallothionein concentrations and decreased the toxicity of cadmium in rats. In vitro the antitoxic activity of methanol extract of H. cordata $T_{HUNB}$ on NIH 3T3 fibroblasts was evaluated by the MTT {3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl-2H-tetrazoliumbromide} and SRB (sulforhodamine B protein) assays. The light microscopic study was carried out to observe morphological changes of the treated cells, $10^{-2}\;mg/ml$ Concentrations of H. cordata $T_{HUNB}$ extract was shown significant antitoxic activity. The number of NIH 3T3 fibroblasts were increased and tend to regenerate. These results suggest that H. cordata $T_{HUNB}$ extract retains a potential antitoxic activity.

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Effects of L-NAME on the Mechanical Hyperalgesia after the Development of Inflammation by Freund's Complete Adjuvant in Rat Paw (FCA에 의한 염증 유발 후 주입된 L-NAME이 기계적 통각과민에 미치는 영향)

  • Kim, Min-Kyung;Choi, Yoon;Kong, Hyun-Seok;Leem, Joong-Woo;Leem, Hang-Soo;Chung, Soo-Jin;Lee, Cheong
    • The Korean Journal of Pain
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    • v.12 no.2
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    • pp.171-176
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    • 1999
  • Background: Effect of nitric oxide on the hyperalgesia induced by inflammation is controversial. From our previous experiment, NOS inhibitor, L-NAME given during the induction period decrease mechanical hyperalgesia occured by Freund's complete adjuvant induced inflammation. In addition, we attempted to analyze the effects of L-NAME on mechanical hyperalgesia after the development of inflammation by Freund's complete adjuvant in rat paw. Methods: Male Sprague Dawley rats were divided into four groups; control (normal saline), and three different doses of L-NAME (0.1 mg, 1 mg, 10 mg). Inflammation was induced in rats by injecting 0.15 ml of Freund's complete adjuvant (FCA) intraplantarly. Rats showed typical hyperalgesia within twelve hours after injection and maintained this for about one week. Tests were done 2 days after injection of FCA. After the baseline test either L-NAME or saline was injected under light halothane anesthesia. Effect of L-NAME on hyperalgesia was assessed by measuring mechanical hyperalgesia at 15, 30, 60, 90, 120 minutes. Same experients were repeated on normal rats. Results: When injected at the site of inflammation, L-NAME caused dose dependent decrease in mechanical hyperalgesia. However, normal rats also showed increased mechanical threshold after L- NAME injection. Conclusions: Although L-NAME reduces FCA induced mechanical hyperalgesia, this result may solely be due to inhibition of nitric oxide production and need to be further determined.

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Effects of Culture Condition on Callus Formation from Anther Culture of Petunia hybrida (Petunia hybrida의 약배양(約培養)으로부터 callus 형성(形成)에 미치는 배양조건(培養條件)의 영향(影響))

  • Chung, Jae Dong;Lee, Jung Hee;Jee, Sun Ok
    • Current Research on Agriculture and Life Sciences
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    • v.10
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    • pp.157-164
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    • 1992
  • The study was conducted to get basic information for haploid production of Petunia hybrida through anther culture by investigating several factors, namely anther stage, culture medium, cold treatment, and genotypes. The results are summarized as follows; Four genotypes of Petunia hybrida were used in a study of anther culture. Plants of each genotype were grown in controlled environments at $20-30^{\circ}C$ with a 16h photoperiod. Equal numbers were harvested from each genotype. The anthers were taken from buds which had received the 14 days' cold treatment at $4^{\circ}C$. Anthers were dissected out aseptically and plated on 1/2 strength MS agar medium containing 5.0mg/${\ell}$ 6-Benzylaminopurine(BAP). Four weeks after culture, light green callus was formed. From these calli, plantlets were regenerated on MS medium containing 2.0mg/${\ell}$ 2-isopentenyl adenine(2ip) after 3 weeks.

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The Research on Pedagogical Content Knowledge(PCK) Focused on Instructional Consulting for Secondary Beginning Teachers (학습자 이해에 관한 초임교사의 수학 수업사례 분석)

  • Choe, Seung-Hyun;Hwang, Hye-Jeang
    • Journal of the Korean School Mathematics Society
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    • v.12 no.4
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    • pp.473-492
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    • 2009
  • Recently there has been a high request for support for teachers' professional development and quality control to meet the demand of educational policy to introduce teacher evaluation, master teacher status, incentives for teacher competency, etc. It has been suggested that reeducation and support for professional development would be more effective to beginning teachers with a high developmental potential than to experienced teachers with routinized instruction. Since 2005, KICE-TLC has conducted research on the development of teacher supporting programs such as teaching consultation and pedagogical content knowledge(PCK) in school subjects. In line with the current education policy and previous research by KICE, this research has been conducted to meet the need for novice teacher induction by developing consulting program focused on PCK. The goal of this research was to (1) explore the in-depth meaning of PCK in light of teaching consultation, (2) conduct a preliminary study on how to develop teaching consulting programs for secondary beginning teachers, (3) develop teaching consulting programs focused on pedagogical content knowledge(PCK), and (4) suggest implications for educational policy regarding pre-service and in-service teachers' continuing professional development and support.

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Significance of Viable but Nonculturable Escherichia coli: Induction, Detection, and Control

  • Ding, Tian;Suo, Yuanjie;Xiang, Qisen;Zhao, Xihong;Chen, Shiguo;Ye, Xingqian;Liu, Donghong
    • Journal of Microbiology and Biotechnology
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    • v.27 no.3
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    • pp.417-428
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    • 2017
  • Diseases caused by foodborne or waterborne pathogens are emerging. Many pathogens can enter into the viable but nonculturable (VBNC) state, which is a survival strategy when exposed to harsh environmental stresses. Pathogens in the VBNC state have the ability to evade conventional microbiological detection methods, posing a significant and potential health risk. Therefore, controlling VBNC bacteria in food processing and the environment is of great importance. As the typical one of the gram-negatives, Escherichia coli (E. coli) is a widespread foodborne and waterborne pathogenic bacterium and is able to enter into a VBNC state in extreme conditions (similar to the other gram-negative bacteria), including inducing factors and resuscitation stimulus. VBNC E. coli has the ability to recover both culturability and pathogenicity, which may bring potential health risk. This review describes the concrete factors (nonthermal treatment, chemical agents, and environmental factors) that induce E. coli into the VBNC state, the condition or stimulus required for resuscitation of VBNC E. coli, and the methods for detecting VBNC E. coli. Furthermore, the mechanism of genes and proteins involved in the VBNC E. coli is also discussed in this review.