• Title/Summary/Keyword: HeLa cell line

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Purification Siderophore from Vibrio mimicus ATCC 33653 and its Effect to Bacterial Pathogenecity (Vibrio mimicus ATCC 33653이 생산하는 Siderophore의 정제와 균의 병원성에 미치는 영향)

  • Park, Soo-Jung;Ju, Seong-A;Heo, Moon-Soo;Jung, Cho-Rok;Ju, Jin-Woo
    • The Journal of the Korean Society for Microbiology
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    • v.34 no.5
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    • pp.461-470
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    • 1999
  • Growth under conditions of iron-restriction and the production of siderophore was examined in Vibrio mimicus ATCC 33653. This strain grew and multiplied in the presence of the high-affinity iron chelators ethylenediamine-di (o-hydroxyphenylacetic acid). Chrome azurol S (CAS) agar and solution were used to detect the production of siderophore under these condition. Siderophore could be detected in the iron-restricted culture supernatants. The siderophore was extracted from iron-restricted culture supernatants by phenol-chloroform-ether method and purified by Dowex ion-exchange and Sephadex G-25 gel filtracton chromatography. The purified siderophore was confirmed by paper chromatography and HPLC. The Purified siderophore enhanced the growth of V. mimicus when the bacterium was grown in iron limited medium. Injection of both the siderohore and the bacteria to mice resulted in more rapid death than that of the only bacteria. However, the siderophore did not show lethality to mice and any toxicity to cell line like HeLa and U937.

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The biological activities of Zelkova leaves and the quality characteristics of Zelkova leaf rice cake according to leaf levels (느티잎의 생리활성 및 느티잎 첨가량에 따른 느티떡의 품질특성)

  • Shon, Jeong-In;Kim, Na-Young;Han, Myung-Joo
    • Korean journal of food and cookery science
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    • v.22 no.4 s.94
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    • pp.468-476
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    • 2006
  • The objectives of this study were to determine the antioxidant and cytotoxic activities of Zelkova leaf extract and the sensory and textural characteristics of Zelkova leaf rice cake. The ethyl acetate layer of Zelkova leaf extract showed the most effective DPPH radical and superoxide anion scavenging activities. The residual layer of Zelkova leaf extract exhibited potent cytotoxicity against the HeLa cell line. In the sensory evaluation, the color and taste of Zelkova leaf rice cakes were not significantly different. Zelkova leaf rice cakes with 20%(5.18), 25%(4.94) and 30%(4.88) added Zelkova leaves showed higher overall acceptability than 0%(4.06) added Zelkova leaves. In the textural analysis, the hardness of the Zelkova leaf rice cake increased slowly with increasing leaf content during 3 day storage. The cohesiveness of 25% and 30% Zelkova leaf rice cakes showed no variation with storage time but that of 0% Zelkova leaf rice cakes decreased with increasing storage period. The 'L' value of Zelkova leaf rice cake decreased with increasing leaf content while the 'a' value of 0% Zelkova leaf rice cake was lower than that of 20%, 25% and 30% Zelkova leaf rice cakes.

Physiological activities of Aronia melanocarpa extracts on extraction solvents (추출용매에 따른 아로니아 추출물의 생리 활성)

  • Park, Hye-Mi;Hong, Joo-Heon
    • Food Science and Preservation
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    • v.21 no.5
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    • pp.718-726
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    • 2014
  • The objective of this study was to investigate the physiological activities of Aronia melanocarpa extracts on extraction solvents (through hot water extraction, 50% ethanol extraction, and 50% methanol extraction). The yield of 50% ethanol extract, 84.50%, was higher than that of the hot water extract (84.05%) and of the 50% methanol extract (76.20%). The total sugar content of the extraction solvent, 35.56~37.68 g/100 g, did not significantly differ. The total anthocyanin content of the 50% methanol extract, 395.10 mg/100 g, was higher than of 50% ethanol extract (318.61 mg/100 g) and of the hot water extract (252.82 mg/100 g). The anthocyanin composition of the cyanidin-3-galactoside, 364.65 mg/100 g, was higher than that of the cyanidin-3-arabinoside (163.06 mg/100 g) and of the cyanidin-3-glucoside (35.69 mg/100 g) in the 50% methanol extract. The DPPH radical scavenging activities of the 50% ethanol and the 50% methanol extracts at $100-1,000{\mu}g/mL$ were 7.96-70.01%, and 8.90-69.21%, respectively. The superoxide radical scavenging activities of all the extracts improved with an increase in the treatment concentration. The FRAP of the 50% ethanol extract and the 50% methanol extract at $100-1,000{\mu}g/mL$ were $57.14-817.87{\mu}M$ and $67.32-812.78{\mu}M$, respectively. The tyrosinase inhibitory activity of the 50% ethanol extract, 23.03-33.82% ($100-1,000{\mu}g/mL$), was higher than that of the other extracts. The cancer cell growth inhibition activity of the 50% ethanol extract (76.86% at $1,000{\mu}g/mL$) on HeLa cell line was significantly higher than of the hot water and of the 50% methanol extracts. There results suggest that the 50% ethanol extract from Aronia melanocarpa may be a useful for functional food material in the food industry.

Investigation of Biological Activities of Enzymatic Hydrolysate of Spirulina (스피루리나 효소가수분해물의 생리활성 탐색)

  • Son, Min-Hee;Park, Keun-Hyoung;Choi, A-Reum;Yoo, Gui-Jae;In, Man-Jin;Kim, Dong-Ho;Chae, Hee-Jeong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.2
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    • pp.136-141
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    • 2009
  • Biological activities of enzymatic hydrolysate of spirulina (EHS) were investigated. EHS showed no significant effects on the growth-stimulating activity for lactic-acid bacteria and antioxidant activity. EHS showed slight in vitro growth-inhibitory effects (15% at 1.42 mg/L) on a human cervical cancer cell line (HeLa). In addition, the anticoagulant activities of EHS were measured based on three different pathways: common, intrinsic and extrinsic pathways. As an indication of anticoagulant activity on common pathway, thrombin time (TT) of EHS (100 mg/L) was measured as 155.6 sec. Activated partial thromboplastin time (aPTT) for intrinsic pathway of EHS (1,000 mg/L) was measured as 95.8 sec. Prothrombin time (PT) based on extrinsic pathway of EHS (1,000 mg/L) was measured as 10.6 sec. These data showed that EHS have influences on anticoagulant factors of common pathway and intrinsic pathway. Consequently it was found that EHS could be used as a functional food for blood circulation.

Antioxidative, Antimutagenic and Cytotoxic Effects of Prunus armeniaca Extracts (살구 추출물의 항산화성, 항돌연변이성 및 세포독성 효과)

  • Yoo, Su-Jung;Kim, Soo-Hyun;Jun, Mi-Sun;Oh, Hyun-Taek;Choi, Hyun-Jin;Ham, Seung-Si
    • Food Science and Preservation
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    • v.14 no.2
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    • pp.220-225
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    • 2007
  • This study was performed to measure the antioxidative, antimutagenic, and cytotoxic properties of Prunus armeniaca using the DPPH (1, 1-diphenyl-2-picrylhydrazyl) free radical donating method, the Ames test, and cytotoxicity measurements, respectively. Electron-donating abilities were 48.3, 43.9, 14.8 and 12.9 per g dry matter of P. armeniaca seed (PAS), P. armeniaca flesh(PAF), butylated hydroxytoluene, and ${\alpha}-tocopherol$, respectively. The direct antimutagenic effects of an ethanol extract of P. armeniaca were examined in Ames tests using Salmonella typhimurium TA98 and TA100 as reporter organisms. In the Ames test, the ethanol extract of P. armenicaca alone did not exhibit any mutagenicity but the extract did show substantial inhibitory effects against mutations induced by N-methyl-N'-nitro-N-nitrosoguanidine(MNNG) and 4-nitroquinoline-1-oxide(4NQO). The ethanol extract of PAS(200g dry matter/plate) inhibited strain TA98 mutagenesis induced by 4NQO by ca. 37.9%, and mutation inhibition values of 42.1% and 69.4%, respectively, were observed when 4NQO and MNNG acted on the TA100 strain. The cytotoxic effects of ethanol extracts of P. armeniaca against cell lines of human lung carcinoma(A549), human breast adenocarcinoma(MCF-7), human hepatocellular carcinoma(Hep3B), human cervical adenocarcinoma(HeLa), and human gastric carcinoma(AGS) rose with increases in extract concentration. An ethanol extract(4mg/mL dry matter) of PAF showed strong cytotoxicities of 88.2%, 58%, 72.8%, 89.4%, and 91.9% against A549, AGS, MCF-7, HeLa, and Hep3B cells, respectively. In contrast, the same extract showed only 13 37% cytotoxicity for a nomal human kiney cell line(293). It is suggested that P. armeniaca possesses useful antioxidative, antimutagenic, and anticancer properties.

Effective Antitumor Activity of a Recombinant Vaccinia Virus Expressing Murine Interleukin 4 (인터루킨-4를 발현하는 재조합 백시니아 바이러스에 의한 암성장의 억제)

  • Yoon, Kee-Jung;Jin, Ning-Yi;Kim, Sun-Young
    • The Journal of Korean Society of Virology
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    • v.28 no.1
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    • pp.71-78
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    • 1998
  • Vaccinia virus is the prototype orthopoxvirus that has been used as a vaccine strain for small pox. This virus has been used to express a variety of cellular and viral genes in mammalian cells at high levels. Interleukin-4 (IL-4) has been found to stimulate the proliferation of T cells and enhance the cytolytic activity of cytotoxic T lymphocytes. To test the immunotherapeutic potential of IL-4 delivered in vivo by poxvirus, a recombinant vaccinia virus expressing the murine IL-4 gene (RVVmIL-4) was constructed. A high level of IL-4 production was confirmed by infecting HeLa cells and measuring IL-4 in cell culture supernatant by ELISA. As a tumor model, two cell lines were used; the murine T leukemic line P388 and the murine breast cancer line TS/A. CDF1 mice were intraperitoneally inoculated with $1\;{\times}\;10^5$ cells of P388. Mice were injected at the same site with $5\;{\times}\;10^5\;PFU$ of recombinant vaccinia virus; first, 3 days after the injection of tumor cells and thereafter once every week for 3 weeks. Intraperitoneal injections of RVVmIL-4 significantly prolonged the survival time of mice inoculated with tumor cells. All mice injected with RVVmIL-4 remained alive for 30 days after the postinoculation of tumor cells, while 100% and 70% of the animals injected with saline or wild type vaccinia virus died, respectively. In another tumor model using TS/A, tumor was established by subcutaneously inoculating $2{\times}10^5$ tumor cells to BALB/c mice. After tumor formation was confirmed on day 4 in all mice, $5\;{\times}\;10^6\;PFU$ of RVVmIL-4 was inoculated subcutaneously three times, once every week for 3 weeks. The TS/A tumor was eradicated in two of the nine mice. Seven of the nine mice treated with RVVmIL-4 developed a tumor, but tumor growth was significantly delayed compared to those treated with saline or wild type vaccinia virus. These results indicate that recombinant vaccinia viruses may be used as a convenient tool for delivering immunomodulator genes to a variety of tumors.

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Immuno-chromatographic Analysis for HPV-16 and 18 E7 Proteins as a Biomarker of Cervical Cancer Caused by Human Papillomavirus

  • Kim, Joo-Ho;Cho, Il-Hoon;Seo, Sung-Min;Kim, Ji-Sook;Oh, Kyu-Ha;Kang, Heun-Soo;Kim, In-Gyu;Paek, Se-Hwan
    • Bulletin of the Korean Chemical Society
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    • v.30 no.12
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    • pp.2999-3005
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    • 2009
  • Among the more than 120 different types of human papillomavirus (HPV), types 16 and 18 have been known to be high risk agents that cause cervical cancer. We examined, in an immuno-chromatographic analysis, the potential of using the early gene product, E7 protein, as a diagnostic marker of cervical cancer caused by HPV. We developed monoclonal antibodies specific to HPV-16 and 18 E7 proteins that were produced from bacterial cells using gene recombinant technology. For each E7 protein, the optimal antibody pair was selected using the immuno-chromatographic sandwichtype binding system based on the lateral flow through membrane pores. Under these conditions, this rapid testing assay had a detection capability as low as 2 ng/mL of E7 protein. Furthermore, since viral analysis required the host cell to be lysed using chemicals such as detergents, it was possible that the E7 protein was structurally damaged during this process, which would result in a decrease in detection sensitivity. Therefore, we examined the detrimental effects caused by different detergents on the E7 protein using HeLa cells as the host. In these experiments, we found that the damage caused by the detergent, nonylphenylpolyethylene glycol (NP-40), was minimal relative to Triton X-100 commonly used for the cell lysis. Temperature also affected the stability of the E7 protein, and we found that the E7 protein was stabilized at 4$^{\circ}C$ for about 2 h, which was 4 times longer than at room temperature. Finally, a HPV-infected cervical cancer cell line, which was used as a real sample model, was treated using the optimized conditions and the presence of E7 proteins were analyzed by immuno-chromatography. The results of this experiment demonstrated that this rapid test could specifically detect HPV-infected samples.

Ethanol but not Aqueous Extracts of Tubers of Sauromatum Giganteum(Engl.) Cusimano and Hett Inhibit Cancer Cell Proliferation

  • Gao, Shi-Yong;Li, Jun;Wang, Long;Sun, Qiu-Jia;Gong, Yun-Fei;Gang, Jian;Su, Yi-Jun;Ji, Yu-Bin
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.24
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    • pp.10613-10619
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    • 2015
  • Background: Both alcohol and aqueous extracts of Sauromatum giganteum(Engl.) Cusimano and Hett, the dried root tuber of which is named Baifuzi in Chinese, have been used for folklore treatment of cancer in Northeast of China. However, little is known about which is most suitable to the cancer therapy. Materials and Methods: Serum pharmacology and MTT assays were adopted to detect the effects of ethanol and aqueous extracts of Sauromatum giganteum(Engl.) Cusimano and Hett, prepared by heat reflux methods, on proliferation of different cancer cells. Results: Cancer cells treated with medium supplemented with 10%, 20%, 40% serum(v/v) containing ethanol extract had a decline in viability, with inhibition rates of 7.69%, 21.8%, 41.9% in MCF-7 cells, 42.8%, 48.1%, 51.8% in SGC-7901 cells, 44.1%, 49.2%, 53.7% in SMMC-7721 cells, 6.8%, 15.2%, 39.8% in HepG2 cells, 7.57%, 16.3%, 36.2% in HeLa cells, 6.24%, 12.5%, 27.4% in A549 cells, and 7.20%, 17.5%, 31.3% in MDA-MB-231 cells, respectively. Viability in the aqueous extract groups was no different with that of controls. Conclusions: An ethanol extract of Sauromatum giganteum(Engl.) Cusimano and Hett inhibited the proliferation of SMMC-7721, SGC-7901 and MCF-7 cells, which supports the use of alcoholic but not aqueous extracts for control of sensive cancers, which might include hepatocarcinoma, gastric cancer and breast cancer.

Safety and Anticancer Effects of Platycodon grandiflorum Extracts (도라지 추출물의 안전성 및 항암 효과)

  • Kim, Soo-Hyun;Chung, Mi Ja
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.4
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    • pp.516-523
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    • 2015
  • This study investigated the antimutagenic and anticancer effects of Platycodon grandiflorum extract (PGE) and its fractions against carcinogenic N-nitrosodimethylamine (NDMA) and genotoxicity. The Ames Salmonella mutagenicity test employing histidine mutants of Salmonella Typhimurium TA98 and TA100 was used to examine the mutagenicity of PGE and its fractions. Bacterial reversion assay with S. Typhimurium TA98 and TA100 did not show a significantly increased number of revertant colonies. The same test was used to examine the ability of PGE and its fractions to prevent acquisition of N-methyl-N'-nitro-N-nitrosoguanidine- and 4-introquino-line-1-oxide-induced mutations. PGE and its fractions inhibited mutagenesis in a dose-dependent manner. Among the fractions, ethyl acetate fraction from PGE (PGEA) exhibited a higher antimutagenic effect than other fractions. PGE and its fractions suppressed the growth of cancer cell lines, including human cervical adenocarcinoma, human hepatocellular carcinoma, human breast adenocarcinoma, human lung carcinoma, and transformed primary human embryonic kidney cells. In addition, we evaluated the antitumor activity of PGEA and its fractions in sacorma-180 solid tumor-bearing mice. In vivo anticancer activity results showed that PGE and its fractions could more effectively suppress tumor growth than the control. PGEA showed higher in vitro and in vivo anticancer effects than PGE and other fractions, and PGEA inhibited NDMA formation. Thus, we showed that PGEA has antimutagenic and anticancer activities, making it a candidate anticancer material under these experimental conditions.

Therapeutic Strategy for the Prevention of Pseudorabies Virus Infection in C57BL/6 Mice by 3D8 scFv with Intrinsic Nuclease Activity

  • Lee, Gunsup;Cho, SeungChan;Hoang, Phuong Mai;Kim, Dongjun;Lee, Yongjun;Kil, Eui-Joon;Byun, Sung-June;Lee, Taek-Kyun;Kim, Dae-Hyun;Kim, Sunghan;Lee, Sukchan
    • Molecules and Cells
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    • v.38 no.9
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    • pp.773-780
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    • 2015
  • 3D8 single chain variable fragment (scFv) is a recombinant monoclonal antibody with nuclease activity that was originally isolated from autoimmune-prone MRL mice. In a previous study, we analyzed the nuclease activity of 3D8 scFv and determined that a HeLa cell line expressing 3D8 scFv conferred resistance to herpes simplex virus type 1 (HSV-1) and pseudorabies virus (PRV). In this study, we demonstrate that 3D8 scFv could be delivered to target tissues and cells where it exerted a therapeutic effect against PRV. PRV was inoculated via intramuscular injection, and 3D8 scFv was injected intraperitoneally. The observed therapeutic effect of 3D8 scFv against PRV was also supported by results from quantitative reverse transcription polymerase chain reaction, southern hybridization, and immunohistochemical assays. Intraperitoneal injection of 5 and $10{\mu}g$ 3D8 scFv resulted in no detectable toxicity. The survival rate in C57BL/6 mice was 9% after intramuscular injection of 10 $LD_{50}$ PRV. In contrast, the 3D8 scFv-injected C57BL/6 mice showed survival rates of 57% ($5{\mu}g$) and 47% ($10{\mu}g$). The results indicate that 3D8 scFv could be utilized as an effective antiviral agent in several animal models.