• Title/Summary/Keyword: HT-29

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Anticancer Activity of Sulfated Polysaccharides Isolated from the Antarctic Red Seaweed Iridaea cordata

  • Kim, Hak Jun;Kim, Woo Jung;Koo, Bon-Won;Kim, Dong-Woo;Lee, Jun Hyuck;Nugroho, Wahyu Sri Kunto
    • Ocean and Polar Research
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    • v.38 no.2
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    • pp.129-137
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    • 2016
  • This study aimed to isolate and characterize sulfated polysaccharides (SPs) from Iridaea cordata and evaluate their anticancer activity. SPs of the Antarctic red seaweed were obtained by $CaCl_2$ (SP1) and ethanol precipitations (SP2) following diluted acid extraction at room temperature. Yields of SP1 and SP2 were approximately 14% and 23%, respectively, of the dry weight of red seaweed. The average molecular mass of the SP1 and SP2 was estimated about $1.84{\times}10^3$ and $1.42{\times}10^3kDa$, respectively, by size-fractionation High-Performance Liquid Chromatography (HPLC). From the High-Performance Anion-Exchange Chromatography-Pulsed Amperometric Detection (HPAEC-PAD) analysis, the main monosaccharide was galactose with glucose and fucose as minor components. The sulfate content of SP2 (40.4%) was slightly higher than that of SP1 (33.8%). The FT-IR spectra also showed characteristic band of carrageenan-like sulfated polysaccharides. Taken together the SPs are thought to be carrageenan-like sulfated galactan. The polysaccharides (SPs) from I. cordata exhibited weak antitumor activity against PC-3 (prostate cancer), HeLa (cervical cancer), and HT-29 (human colon adenocarcinoma). To our knowledge, this is the first data on biological activity of the Antarctic red seaweed I. cordata.

Effects of Aralia continentalis Root Extract on Cell Proliferation and Apoptosis in Human Promyelocytic Leukemia HL-60 Cells

  • Lim Hae-Young;Oh Ha-Lim;Lee Chul-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.16 no.9
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    • pp.1399-1404
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    • 2006
  • The roots of Aralia continentalis (AC) have been used traditionally in Korean as a folk medicine for anti-inflammation and as an anti-rheumatic. In this study, we report that the ethyl acetate-soluble traction (ACE) of the methanolic extract of AC root inhibited the cell growth of various human cancer cell lines and induced apoptosis of HL-60, human promyelocytic leukemia cells. Its $IC_{50}$ values on growth inhibition were estimated to be $56.3{\mu}g/ml$ on HL-60, $87.2{\mu}g/ml$ on HepG2, $93.2{\mu}g/ml$ on HeLa, $135.5{\mu}g/ml$ on DU-145, and $135.8{\mu}g/ml$ on HT-29 cells. Interestingly, ACE showed no antiproliferative effect on normal lymphocyte cells used as control. Furthermore, nuclear DAPI staining revealed the typical nuclear features of apoptosis in the HL-60 cells exposed to $80{\mu}g/ml$ ACE, and a flow cytometric analysis of the HL-60 cells using propidium iodide showed that the apoptotic cell population increased gradually from 5% at 0 h to 16% at 12 h and 20% at 24 h after treated with $50{\mu}g/ml$ of ACE. TUNEL assay also revealed the apoptotic induction of the HL-60 cells treated with ACE. To obtain further information on the ACE-induced apoptosis, the expression level of certain apoptosis-associated proteins was examined using a Western blot analysis. Treatment of the HL-60 cells with ACE resulted in the activation of caspase-3, and subsequent proteolytic cleavage of poly(ADP-ribose) polymerase (PARP). The above results confirmed that the apoptosis in the HL-60 cells was induced by ACE, and that caspase-3-mediated PARP cleavage was involved in the process.

Physico-chemical Properties and In Vivo Anti-cancer Effects of Potato Kimchi Prepared by adding Hot Water Extracts of Potato (생감자의 열탕 추출물을 첨가한 감자 김치의 이화학적 특성 및 In Vivo에서의 항암 효과)

  • Chang, Sang-Keun;Kim, Hee-Joo
    • Journal of the East Asian Society of Dietary Life
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    • v.18 no.3
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    • pp.302-310
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    • 2008
  • In this study, potato kimchi was prepared by applying heat to raw potatoes, and then the physico-chemical properties and anti-cancer effects of the kimchi were analyzed. The texture results indicated the potato kimchi had very good hardness and springiness attributes. During th late storage period, total vitamin C content of the kimchi slowly increased. In addition, the potato kimchi had non-volatile organic acid changes that promoted early aging; however, after the complete aging period, it was comparatively similar to other types of kimchi. Using the methanol extracts of various kimchi samples, the potato kimchi(solid 100%) showed the highest anti-carcinogenic effects in terms of anti-tumor activity in tumor bearing Balb/c mice with sarcoma-180 cells. In addition, the effects of the methanol extracts on hepatic glutathione S-transferase content were $289.76\;{\mu}mol/mg$ protein/min, $250.97\;{\mu}mol/mg$ protein/min, $251.20\;{\mu}mol/mg$ protein/min, $219.53\;{\mu}mol/mg$ protein/min, $183.79\;{\mu}mol/mg$ protein/min, for control kimchi, mul kimchi, and two potato kimchis [(solid 100%) and(solid 60%+kimchi juice 40%)], respectively. The in vivo anti-cancer effects of the potato kimchi were investigated using AGS human gastric adenocarcionoma cells and HT-29 human colon adenocarcionoma cells. Overall, an MTT assay revealed that the methanol extract of the potato kimchi showed the highest anti-carcinogenic effects.

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Clostridium difficile Toxin A Inhibits the Kinase Activity of Extracellular Signal-Related Kinases 1 and 2 Through Direct Binding

  • Seok, Heon;Nam, Hyo-Jung;Nam, Seung-Taek;Kang, Jin-Ku;Kim, Sung-Kuk;Chang, Jong-Soo;Ha, Eun-Mi;Park, Young-Joo;Kim, Ho
    • Journal of Microbiology and Biotechnology
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    • v.22 no.2
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    • pp.170-175
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    • 2012
  • Clostridium difficile toxin A glucosylates Rho family proteins, resulting in actin filament disaggregation and cell rounding in cultured colonocytes. Given that the cellular toxicity of toxin A is dependent on its receptor binding and subsequent entry into the cell, we herein sought to identify additional colonocyte proteins that might bind to toxin A following its internalization. Our results revealed that toxin A interacted with ERK1 and ERK2 in two human colonocyte cell lines (NCM460 and HT29). A GST-pulldown assay also showed that toxin A can directly bind to ERK1 and ERK2. In NCM460 cells exposed to PMA (an ERK1/2 activator), the phosphorylation of ERK1/2 did not affect the interaction between toxin A and ERK1/2. However, an in vitro kinase assay showed that the direct binding of toxin A to ERK1 or ERK2 inhibited their kinase activities. These results suggest a new molecular mechanism for the cellular toxicity seen in cells exposed to toxin A.

Isolation and characterization of cholesterol-lowering lactic acid bacteria from kimchi (김치에서 분리된 콜레스테롤 감소능을 가진 젖산세균의 특성)

  • Park, Hong-Yeop;Park, Seul-Ki;Kim, Bo-Geum;Ryu, Dae-Gyu;Lim, Eun-Seo;Kim, Young-Mog
    • Korean Journal of Food Science and Technology
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    • v.49 no.4
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    • pp.377-382
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    • 2017
  • The objective of this study was to isolate and characterize lactic acid bacteria (LAB) exhibiting cholesterol-lowering activity from the Korean traditional fermented food, kimchi. The previously isolated LAB strains were assessed for cholesterol-lowering efficacy in the presence of 0.1% cholesterol. All LAB strains tested in this study were able to assimilate cholesterol at varying levels, ranging from 35.0 to 99.4%. Among them, the Lactobacillus plantarum FMB 31 strain exhibited the highest cholesterol-lowering effect with 99.4% cholesterol removal efficiency. The strain was stable in the presence of acid, bile, and salt stress, and showed high adherence on HT-29 cells, a human colon line. In addition, the LAB strain showed no pathogenic properties such as the production of hemolysin and biogenic amines. Thus, this study suggests that the L. plantarum FMB 31 strain isolated from kimchi can be a potential source of probiotic products with strong cholesterol-lowering effect.

Comparative Genomic Analysis of Lactobacillus rhamnosus BFE5264, a Probiotic Strain Isolated from Traditional Maasai Fermented Milk

  • Jeong, Haeyoung;Choi, Sanghaeng;Park, Gun-Seok;Ji, Yosep;Park, Soyoung;Holzapfel, Wilhelm Heinrich;Mathara, Julius Maina;Kang, Jihee
    • Microbiology and Biotechnology Letters
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    • v.47 no.1
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    • pp.25-33
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    • 2019
  • Lactobacillus rhamnosus BFE5264, isolated from a Maasai fermented milk product ("kule naoto"), was previously shown to exhibit bile acid resistance, cholesterol assimilation, and adhesion to HT29-MTX cells in vitro. In this study, we re-annotated and analyzed the previously reported complete genome sequence of strain BFE5264. The genome consists of a circular chromosome of 3,086,152 bp and a putative plasmid, which is the largest one identified among L. rhamnosus strains. Among the 2,883 predicted protein-coding genes, those with carbohydrate-related functions were the most abundant. Genome analysis of strain BFE5264 revealed two consecutive CRISPR regions and no known virulence factors or antimicrobial resistance genes. In addition, previously known highly variable regions in the genomes of L. rhamnosus strains were also evident in strain BFE5264. Pairwise comparison with the most studied probiotic strain L. rhamnosus GG revealed strain BFE5264-specific deletions, probably due to insertion sequence-mediated recombination. The latter was associated with loss of the spaCBA pilin gene cluster and exopolysaccharide biosynthetic genes. Comparative genomic analysis of the sequences from all available L. rhamnosus strains revealed that they were clustered into two groups, being within the same species boundary based on the average nucleotide identities. Strain BFE5264 had a sister group relationship with the group that contained strain GG, but neither ANI-based hierarchical clustering nor core-gene-based phylogenetic tree construction showed a clear distinctive pattern associated with the isolation source, implying that the genotype alone cannot account for their ecological niches. These results provide insights into the probiotic mechanisms of strain BFE5264 at the genomic level.

Actinidia arguta Sprout as a Natural Antioxidant: Ameliorating Effect on Lipopolysaccharide-Induced Cognitive Impairment

  • Kang, Jeong Eun;Park, Seon Kyeong;Kang, Jin Yong;Kim, Jong Min;Kwon, Bong Seok;Park, Sang Hyun;Lee, Chang Jun;Yoo, Seul Ki;Heo, Ho Jin
    • Journal of Microbiology and Biotechnology
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    • v.31 no.1
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    • pp.51-62
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    • 2021
  • Here, we investigated the prebiotic and antioxidant effects of Actinidia arguta sprout water extract (AASWE) on lipopolysaccharide (LPS)-induced cognitive deficit mice. AASWE increased viable cell count, titratable acidity, and acetic acid production in Lactobacillus reuteri strain and showed a cytoprotective effect on LPS-induced inflammation in HT-29 cells. We assessed the behavior of LPS-induced cognitive deficit mice using Y-maze, passive avoidance and Morris water maze tests and found that administration of AASWE significantly improved learning and memory function. The AASWE group showed antioxidant activity through downregulation of malondialdehyde levels and upregulation of superoxide dismutase levels in brain tissue. In addition, the AASWE group exhibited activation of the cholinergic system with decreased acetylcholinesterase activity in brain tissue. Furthermore, AASWE effectively downregulated inflammatory mediators such as phosphorylated-JNK, phosphorylated-NF-κB, TNF-α and interleukin-6. The major bioactive compounds of AASWE were identified as quercetin-3-O-arabinopyranosyl(1→2)-rhamnopyranosyl(1→6)-glucopyranose, quercetin-3-O-apiosyl(1 → 2)-galactoside, rutin, and 3-caffeoylquinic acid. Based on these results, we suggest that AASWE not only increases the growth of beneficial bacteria in the intestines, but also shows an ameliorating effect on LPS-induced cognitive impairment.

Cr Electroplating Technology to prevent Interdiffusion between Metallic Fuel and Clad Material (금속연료-피복재 상호확산 방지를 위한 크롬 도금법 적용 연구)

  • Kim, Jun Hwan;Lee, Kang Soo;Yang, Seong Woo;Lee, Byoung Oon;Lee, Chan Bock
    • Korean Journal of Metals and Materials
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    • v.49 no.12
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    • pp.937-944
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    • 2011
  • Studies have been carried out in order to reduce fuel-cladding chemical interaction (FCCI) behavior of metallic fuel in sodium-cooled fast reactors (SFR) using an electroplating technique. A $20{\mu}m$ thick Cr layer has been plated by the electrochemical method in the Sargent bath over the HT9 (12Cr-1Mo) clad material and diffusion couple tests of the U-10Zr metallic fuel as well as the rare earth alloy (70Ce-29La) have been conducted. The results show that the Cr plating can prevent FCCI behavior along the fuel-clad interface. However, cracks developed through the thickness during plating, which resulted in the migration of some fuel constituents. Variation of bath temperature, application of pulse current, and post heat treatment have been conducted to control such cracks. We found out that some conditions like the pulse current and the post heat treatment enhanced the layer property by reducing the internal cracks and improving the diffusion couple test.

β-carotene regulates cancer stemness in colon cancer in vivo and in vitro

  • Lee, Kyung Eun;Kwon, Minseo;Kim, Yoo Sun;Kim, Yerin;Chung, Min Gi;Heo, Seung Chul;Kim, Yuri
    • Nutrition Research and Practice
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    • v.16 no.2
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    • pp.161-172
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    • 2022
  • BACKGROUND/OBJECTIVES: Colorectal cancer (CRC) is the third most common cancer worldwide and has a high recurrence rate, which is associated with cancer stem cells (CSCs). β-carotene (BC) possesses antioxidant activity and several anticancer mechanisms. However, no investigation has examined its effect on colon cancer stemness. MATERIALS/METHODS: CD133+CD44+ HCT116 and CD133+CD44+ HT-29 cells were isolated and analyzed their self-renewal capacity by clonogenic and sphere formation assays. Expressions of several CSCs markers and Wnt/β-catenin signaling were examined. In addition, CD133+CD44+ HCT116 cells were subcutaneously injected in xenograft mice and analyzed the effect of BC on tumor formation, tumor volume, and CSCs markers in tumors. RESULTS: BC inhibited self-renewal capacity and CSC markers, including CD44, CD133, ALDH1A1, NOTCH1, Sox2, and β-catenin in vitro. The effects of BC on CSC markers were confirmed in primary cells isolated from human CRC tumors. BC supplementation decreased the number and size of tumors and delayed the tumor-onset time in xenograft mice injected with CD133+CD44+ HCT116 cells. The inhibitory effect of BC on CSC markers and the Wnt/β-catenin signaling pathway in tumors was confirmed in vivo as well. CONCLUSIONS: These results suggest that BC may be a potential therapeutic agent for colon cancer by targeting colon CSCs.