• Title/Summary/Keyword: cell fractions

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Physicochemical Properties and Intestinal Bacterial Growth-Promoting Effect of Cell-Wall Polysaccharides from Cucumber Peel

  • Jun, Hyun-Il;Song, Geun-Seoup;Lee, Young-Tack;Kim, Young-Soo
    • Food Science and Biotechnology
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    • v.14 no.3
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    • pp.375-379
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    • 2005
  • Physicochemical properties, intestinal microbial growth, and inhibitory effects of alcohol-insoluble polysaccharide (AIP) extracted from cucumber peel were investigated. AIP was composed of 14.54% crude protein, 1.04% crude lipid, 13.74 % crude ash, 9.1% soluble dietary fiber, and 41.2% insoluble dietary fiber. AIP showed low bulk density (0.18 g/mL) and water-holding capacity (6.39 g/g), and high oil-holding capacity (3.96 g/g). Pectic substance fractions [water-soluble pectic substance (WSP), ethylenediaminetetraacetic acid-soluble pectic substance (ESP), and alkali-soluble pectic substances (ASP)] and hemicellulose fractions [1 M KOH-soluble hemicellulose (KHP1) and 4 M KOH-soluble hemicellulose (KHP4)] were obtained from sequential chemical fractionation of AIP. WSP showed higher total sugar contents than total uronic acid contents, whereas opposite results were observed in ESP and ASP. Molecular weight distributions of three pectic substance fractions were in order of ASP>ESP>WSP. Ion exchange chromatogram pattern of WSP was different from those of ESP and ASP. Major component of WSP was fraction eluted by 0.05 M ammonium acetate buffer, whereas that of ESP and ASP was fraction eluted by 0.2 M NaOH. WSP and ASP showed growth-promoting activities against Lactobacillus brevis, Bifidobacterium bifidum, and B. longum, whereas B. bifidum and B. longum for ESP. KHP1 and KHP4 fractions had significant growth-promoting activities against B. bifidum.

Chemical Components and Biological Activities of Red Onion Powder (붉은 양파 분말의 화학성분 및 생리활성)

  • Jang, Joo-Ri;Kwon, Sun-Jin;Lim, Sun-Young
    • Journal of the Korean Society of Food Culture
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    • v.24 no.6
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    • pp.749-755
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    • 2009
  • We investigated the chemical components of red onion powder dried using the low temperature vacuum method and the inhibitory effects of solvent extracts of the dried red onion powder on the growth of HT-1080 human fibrosarcoma and HT-29 human colon cancer cells and $H_2O_2$-induced oxidative stress. The moisture content of the dried red onion powder was 17.95%, while the vitamin C content was 96 mg/100 g and the total phenols content was 39.1 mg/mL. The inhibitory effects of acetone with methylene chloride (A+M) and methanol (MeOH) extracts of the red onion powder on the growth of HT-1080 and HT-29 cancer cells increased in a dose dependent manner (p<0.05). The inhibitory effect was greater on the growth of HT-29 cells, while the A+M extracts had a higher inhibitory effect than the MeOH extracts. Treatment with the hexane, 85% aq. methanol, butanol and water fractions of the extract led to significant inhibition of the growth of both cancer cell lines (p<0.05). Among the fractions, the hexane and 85% aq. methanol fractions showed a greater inhibitory effect. To determine the protective effect on $H_2O_2$-induced oxidative stress, a DCFH-DA (dichlorodihydrofluorescin diacetate) assay was conducted. All fractions, including the crude extracts of dried red onion, appeared to lead to a significant reduction in the levels of intracellular reactive oxygen species (ROS), and these reductions occurred in a dose dependent fashion (p<0.05). Among the fractions, the 85% methanol fraction showed the greatest protective effect on the production of lipid peroxides.

In vitro studies of anti-inflammatory and anticancer activities of organic solvent extracts from cultured marine microalgae

  • Samarakoon, Kalpa W.;Ko, Ju-Young;Shah, Md. Mahfuzur Rahman;Lee, Ji-Hyeok;Kang, Min-Cheol;Kwon, O-Nam;Lee, Joon-Baek;Jeon, You-Jin
    • ALGAE
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    • v.28 no.1
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    • pp.111-119
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    • 2013
  • Marine microalgae are a promising source of organisms that can be cultured and targeted to isolate the broad spectrum of functional metabolites. In this study, two species of cyanobacteria, Chlorella ovalis Butcher and Nannchloropsis oculata Droop, one species of bacillariophyta, Phaeoductylum tricornutum Bohlin, and one species of Dinophyceae, Amphidinium carterae (Hulburt) were cultured and biomasses used to evaluate the proximate comical compositions. Among the determined proximate chemical compositions of the cultured marine microalgae, the highest content of crude proteins and lipids were exhibited in P. tricornutum and A. carterae, respectively. Solvent-solvent partition chromatography was subjected to fractionate each of the cultured species and separated n-hexane, chloroform, ethyl acetate, and aqueous fractions. Nitric oxide production inhibitory level (%) and cytotoxicity effect on lipo-polysaccharide-induced RAW 264.7 macrophages were performed to determine the anti-inflammatory activity. N. oculata hexane and chloroform fractions showed significantly the strongest anti-inflammatory activity at $6.25{\mu}g\;mL^{-1}$ concentration. The cancer cell growth inhibition (%) was determined on three different cell lines including HL-60 (a human promyelocytic leukemia cell line), A549 (a human lung carcinoma cell line), and B16F10 (a mouse melanoma cell line), respectively. Among the extracts, C. ovalis ethyl acetate and A. carterae chloroform fractions suppressed the growth of HL-60 cells significantly at 25 and $50{\mu}g\;mL^{-1}$ concentrations. Thus, the cultured marine microalgae solvent extracts may have potentiality to isolate pharmacologically active metabolites further using advance chromatographic steps. Hence, the cultured marine microalgae can be described as a good candidate for the future therapeutic uses.

Growth Inhibitory and Quinone Reductase Activity Stimulating Effects of Internal Organs of Todarodes pacificus Fractions on Human Cancer Cell Lines In vitro (오징어내장 분획물의 in vitro에서의 암세포 성장억제 및 quinone reductase유도 활성 증가 효과)

  • Shin, Mi-Ok;Bae, Song-Ja
    • Journal of Life Science
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    • v.19 no.9
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    • pp.1251-1257
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    • 2009
  • We investigated the growth inhibitory effect of internal organs of Todarodes pacificus (TP) on proliferation in human cancer cell lines in vitro. The internal organs of TP were extracted with methanol (TPM), which was then further fractionated into four subfractions by using a solvent partition method, resulting in hexane (TPMH), methanol (TPMM), butanol (TPMB), and aqueous (TPMA) soluble fractions. We determined the cytotoxic effect of these four fractions in three kinds of cancer cell lines - HepG2, MCF-7 and HT-29 - by MTT assay. Among the four subfractions of TPM, TPMH showed the strongest cytotoxic effects at a concentration of $300{\mu}g$/ml, displaying 91.56% on the HepG2 cell line and 85.93% on the MCF-7 cell line. Morphological changes such as membrane shirinking and blebbing of cells were also observed during TPMH treatment of HepG2 cells. In addition, we also observed quinone reductase (QR) induced effect in the methanol (TPMM) layer of HepG2 cells. TPMM showed the highest induction activity of quinone reductase on HepG2 cells among the other partition layers. The QR induced effect of TPMM was determined to be 2.7 at a level of $360{\mu}g$/ml with a control value of 1.0. Although further studies are needed, the present work suggests that internal organs of Todarodes pacificus (TP) may be a chemopreventive agent for the treatment of human cells.

All-trans Retinoic Acid Induced Differentiation of Rat Mammary Epithelial Cells Cultured in Serum-free Medium

  • Ki, Min-Hyo;Paik, Kee-Joo;Lee, Ji-Hyeon;Chung, Hae-Young;Lee, Kyung-Hee;Kim, Kyu-Won;Kim, Nam-Deuk
    • Archives of Pharmacal Research
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    • v.21 no.3
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    • pp.298-304
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    • 1998
  • Retinoids are applied to not only cancer prevention but also cancer chemotherapy by stimulating differentiation of cells. We studied differentiation inducing effect of all-trans retinoic acid (ATRA) by studying proportion of high dense fractions of stem-like cells and the size of S phase fraction in cell cycle. From mammary organoids obtained from 7- to 8-week old F344 female rat mammary gland, we cultured rat mammary epithelial cells (RMEC) and treated physiological doses of $10^{-6}$, $10^{-7}$, and $10^{-8}$ M ATRA from the first day and then cultured for 4, 7, and 14 days. After that, immunostaining was performed using peanut agglutinin (PNA) and anti-Thy-1.1 monoclonal antibody (Thy-1.1) that can be used as markers of differentiation. We separated four different cell subpopulations by flow cytometry: cells negative to both reagents (B-), PNA-positive cells (PNA+), Thy-1.1-positive cells (Thy-1.1+), and cells positive to both reagents (B+). We observed continuous decreases of high dense fractions of stem-like cells (PNA+ subpopulations) for 14 days and as much decreases as high doses of ATRA, which were thought to be proportional to doses of ATRA. We labeled RMEC with bromodeoxyuridine and investigated cell cycle fractions that went through S phase. We observed a tendency of decrease of S phase fraction with time in culture, which, is thought to be related to continuous decreases of PNA+ subpopulations and inhibitory role of ATRA on cell cycle. These results suggest that physiological doses of ATRA could stimulate differentiation of RMEC and convert stem-like RMEC to differentiated cells in SFM for a relatively long period of 14 days.

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Relationship between Radiosensitivity and Repair Capacity in Human Epithelial Cancer Cell Lines (인체 상피암 세포주에서 방사선감수성과 손상회복의 상관관계에 관한 연구)

  • Koh, Kyoung-Hwan;Ha, Sung-Whan;Park, Charn-Il
    • Radiation Oncology Journal
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    • v.11 no.1
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    • pp.17-27
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    • 1993
  • To investigate the relationship between radiosensitivity and postirradiation recovery in human cancer cells, a study was performed using human cancer cell lines-A549, CaSki, SNU-C5 and PCI-13. For the study of radiosensitivity, single doses of 2, 4, 6, 8, 10, 12, and 14 Gy were given and for postirradiation recovery, two fractions of 4 Gy were separated with a time interval of 0, 0.5, 1, 1.5, 2, 2.5, 3, 4, 5, or 6 hours. Surviving fraction was estimated using colony forming ability. Surviving fractions at 2 Gy (SF2) were 0.496 (0.570-0.412) for A549, 0.496 (0.660-0.332) for CaSki, 0.386 (0.576-0.216) for SNU-C5, and 0.185 (0.247-0.123) for PCI-13. By statistical analysis the SF2 of PCI-13 was lower significantly than those of others (p<0.05). This difference was also observed at 4, 6 and 8 Gy dose levels. At 6 and 8 Gy the surviving fractions of SNU-C5 were also lower significantly than A549 and CaSki (p<0.05). By the analysis with linear quadratic model, the values of ${\alpha}$ for A549, CaSki, SNU-C5 and PCI-13 were 0.3016, 0.3212, 0.4327 and 0.8423, respectively, and those of ${\beta}$ were 0.02429, 0.02009, 0.03349 and 0.00059, respectively. So, the value of ${\alpha}$ showed increasing tendency with decreasing SF2. By the multitarget single hit model the values of Do for A549, CaSki, SUN-C5 and PCI-13 were 1.97, 1.97, 1.46 and 0.81, respectively, and those of n were 1.53, 1.50, 1.56 and 2.28, respectively. So, the value of Do decreased with decreasing SF2. Post-irradiation recovery reached plateau at around 2 hours. Recovery ratio at plateau phase ranged from 1.2 to 4.2; the value were 1.2 for PCI-13, 3.2 for CaSki, 3.3 for SNU-C5, and 4.2 for A549. Recovery rate well correlated with SF2, and increased with increasing Do and decreasing ${\alpha}$. According to above results, the intrinsic radiosensitivity was quite different among the tested cell lines; PCI-13 was the most sensitive and A549 and CaSki was similar. This difference of radiosensitivity is thought to be partly due to the difference in amount of postirradiation recovery. By linear quadratic model the difference of ${\alpha}$ values was very high, and by multitarget single hit model the difference of Do value was significantly high among four cell lines.

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Antioxidant Activity of Solvent Fractions from Distylium racemosum in Jeju (제주 자생 조록나무 분획물의 항산화 효과)

  • Kim, Hye-Ran;Park, Gyu-Nam;Jung, Bo-Kyoung;Yoon, Weon-Jong;Jung, Yong-Hwan;Chang, Kyung-Soo
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.2
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    • pp.62-67
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    • 2016
  • Anti-oxidant activity of 600 medicinal plants from Jeju was analyzed. Extracts from the leaves of Distylium racemosum have the highest anti-oxidant activity. D. racemosum is an oriental medicinal plant belonging to the Hamamelidacea and grows in the wild in Jeju. This study was conducted to evaluate the antioxidant and cell viability of different fractions (n-hexane, methylene chloride, ethyl acetate, buthanol, DW) from D. racemosum. Anti-oxidant activity was measured using 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity test and total phenolic content. Cell viability was determined by 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) cell viability assay on HepG2 and A549 cells. Among various extracts, the ethyl acetate fraction showed the highest DPPH radical scavenging activity, reaching approximately 93% at 0.5 mg/mL, higher than that of quercetin used as a positive control. Ethyl acetate fractions showed the highest total phenolic content at 505 mg GAE/g. The phenolic content of each extract showed association with DPPH radical scavenging activity. The ethyl acetate extracts were resistant against hydrogen peroxide ($H_2O_2$) treatment in the MTT cell viability assay and showed a higher cell protective effect than other fraction extracts. These results suggest that the ethyl acetate fraction might be a source of anti-oxidants of D. racemosum.

Characterization and Action Mode of Anti-Complementary Substance Prepared from Lactobacillus plantarum (Lactobacillus plantarum 균체 중 항보체 활성물질의 특성과 작용양식)

  • Kim, Jang-Hyun;Shin, Kwang-Soon;Lee, Ho
    • Korean Journal of Food Science and Technology
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    • v.34 no.2
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    • pp.290-295
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    • 2002
  • Among 12 lactic acid bacteria examined for their abilities to activate the complement system by hemolytic complement assay $(TCH_{50})$, Lactobacillus plantarum previously isolated from Kimchi showed high anti-complementary activity. The anti-complementary activity of the cell wall fraction of L. plantarum was more potent than that of the cytosol fraction, and both activities showed dose dependency. These high activities of the cytosol and the cell wall fractions were relatively resistant to the digestion with pronase, but sharply decreased after the treatment of $NaIO_4$. These results suggested that the complement activation by the cytosol and the cell wall fractions was mainly due to their polysaccharides. By the cross-immunoelectrophoresis using anti-human C3, the C3 activation products from both fractions were identified in $Ca^{++}$-free condition. Anti-complementary activity $(ITCH_{50})$ of the cell wall fraction was retained under the same condition, whereas that of the cytosol fraction was reduced considerably. From these results, it was inferred that the mode of complement activation by the cell wall fraction was mainly via alternative pathway, and that of the cytosol fraction was via both alternative and classical pathways.

Growth Inhibitory and Quinone Reductase Induction Activities of Salicornia herbacea L. Fractions on Human Cancer Cell Lines in vitro (함초 분획물의 in vitro에서의 암세포 성장억제 및 Quinone Reductase 활성 유도 효과)

  • Jung, Bok-Mi;Park, Jung-Ae;Bae, Song-Ja
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.2
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    • pp.148-153
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    • 2008
  • We investigated the growth inhibitory effect of Salicornia herbacea L. (SH) on human cancer cell lines in vitro. SH was extracted with methanol (SHM), followed by further fractionation into four subfractions according to polarity: hexane (SHMH), methanol (SHMM), butanol (SHMB), and aqueous (SHMA) soluble fractions. We determined the growth inhibitory effect of these fractions against human cancer cell lines using MTT assay. Among the four subfractions of SHM, the SHMM showed the strongest cytotoxic effects on cancer cell lines. We also observed quinone reductase (QR)-inducing effect of methanol layer (SHMM) on HepG2 cells and it was determined to be 3.00 at $100\;{\mu}g/mL$ level compared to the control value of 1.0. The SHMM showed the highest induction activity of quinone reductase on HepG2 cells among the partition layers. The present work suggests that SH merits further study to confirm its chemopreventive potential.

Anti-oxidative Activities for the Flavonoids of the Syzygium aqueum Burm.f. Alston Branches from Jeju Island (제주 자생 물사과 가지 유래 Flavonoid 화합물의 항산화 활성)

  • Yeom, Hyun Sook;Lee, Nam Ho;Hyun, Ju Mi
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.44 no.2
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    • pp.151-159
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    • 2018
  • In this study, we investigated the anti-oxidative activities and cell protective effects of the constituents isolated from S. aqueum branches. DPPH and $ABTS^+$ radical scavenging activities were screened for the ethanol extract and solvent fractions, ethyl acetate (EtOAc) and butanol (BuOH) fractions showed potent activities. When HaCaT cells were treated with $H_2O_2$, the ethanol extract and EtOAc fractions ($20{\mu}g/mL$) protected the cells against oxidative damage. Two constituents were isolated from the EtOAc fraction of S. aqueum branches; pinocembrin (1), desmethoxymatteucinol (2). The chemical structures of the isolated compounds were elucidated based on the spectroscopic data including $^1H$ and $^{13}C$ NMR spectra, as well as comparison of the data to the literature values. Anti-oxidative activities and cell protective effects were studied for the isolated compounds. For the anti-oxidative activities, all of the compounds 1 and 2 showed DPPH and $ABTS^+$ radical scavenging activities. Also, from the cell protective effect test, the compounds 1 and 2 protected the cell against oxidative stress by $H_2O_2$. Based on these results, S. aqueum branches extract could be potentially applicable as anti-oxidant ingredients in cosmetic industries.