• Title/Summary/Keyword: CR2 cells

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Methed for the Passaging of Microcarrier Cultures to a Production Scale for Producing High Titre Disabled Infectious Single Cycle-Herpes Simplex virus Type-2

  • Zecchini, Tracey-Ann;Wright, Paul-Andrew;Smith, Rodney-John
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.2
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    • pp.118-122
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    • 2000
  • A comlementary call line CR2 is curretly used to propagte the Disabled Infectious Single Cycle Herpes Simplex Virus Typee2 (DISC HSV-2) on a small Iaboratory scale upto 15 L. These cultures are initiated by passaging the cells from roller bottle cultures. Whilst this is suitable for the laboratory scale it is totally impractical for use in seeding an industrial manufacturing scaled version of the culture. It is paramount to have a robust system for passaging cells from a small microcarrierier culture system to a larger one by a serial subculturing regime. Here we report on the successes we have had in our laboratory in scaling up out production system for the DISC HSV-2 from small 1-L cultures to a 50-L vessel with the maintenance of the viral productivity. Ease of use, reproducibility and the need to minimise overall production time were factors which were taken into consideration whils developing our procedures. We were aware of the need to keep a production train simple and as short as possible as this was the amall scale study for an envisaged manufacturing process.

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Effects of Antidiabetic Agent, Aloe QDM complex, on Intracellular Glucose Uptake (항당뇨 물질 Aloe QDM complex의 세포내 포도당 흡수촉진 효능)

  • Im, Sun-A;Kim, Ki-Hyang;Shin, Eunju;Do, Seon-Gil;Jo, Tae Hyung;Park, Young-In;Lee, Chong-Kil
    • Korean Journal of Pharmacognosy
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    • v.44 no.1
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    • pp.75-82
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    • 2013
  • Previous studies have shown that Aloe QDM complex, which is consisted of chromium (Cr), aloesin (ALS) and processed Aloe vera gel (PAG), exert antidiabetic activity in a high fat diet-induced mouse model of type 2 diabetes. In this study we examined the mechanism of the antidiabetic activity of the Aloe QDM complex. Rat myoblast cell line L6 cells were cultured in the presence of Cr, ALS, and PAG alone and in combinations, and then the capability of the cells to uptake glucose was examined using radiolabeled glucose. All of the 3 agents, Cr, ALS and PAG, exerted glucose uptake-enhancing activity in L6 cells. The most potent capability to uptake glucose was observed when L6 cells were cultured with the Aloe QDM complex. The activity of the Aloe QDM complex to enhance glucose uptake was prominent in conditions where existing insulin concentrations are low. We also examined the effects of the Aloe QDM complex on the plasma membrane expression of GLUT4 in L6 cells. The Aloe QDM complex increased the content of GLUT4 in the plasma membrane, while decreasing the content of GLUT4 in the light microsome. Taken together, these results show that the antidiabetic activity of the Aloe QDM complex is at least in part due to the stimulation of glucose uptake into the muscle cells, and this activity of the Aloe QDM complex is mediated through the enhancement of the translocation of GLUT4 into the plasma membrane.

CrABCA2 Facilitates Triacylglycerol Accumulation in Chlamydomonas reinhardtii under Nitrogen Starvation

  • Jang, Sunghoon;Kong, Fantao;Lee, Jihyeon;Choi, Bae Young;Wang, Pengfei;Gao, Peng;Yamano, Takashi;Fukuzawa, Hideya;Kang, Byung-Ho;Lee, Youngsook
    • Molecules and Cells
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    • v.43 no.1
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    • pp.48-57
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    • 2020
  • The microalga Chlamydomonas reinhardtii accumulates triacylglycerols (TAGs) in lipid droplets under stress conditions, such as nitrogen starvation. TAG biosynthesis occurs mainly at the endoplasmic reticulum (ER) and requires fatty acid (FA) substrates supplied from chloroplasts. How FAs are transferred from chloroplast to ER in microalgae was unknown. We previously reported that an Arabidopsis thaliana ATP-binding cassette (ABC) transporter, AtABCA9, facilitates FA transport at the ER during seed development. Here we identified a gene homologous to AtABCA9 in the C. reinhardtii genome, which we named CrABCA2. Under nitrogen deprivation conditions, CrABCA2 expression was upregulated, and the CrABCA2 protein level also increased. CrABCA2 knockdown lines accumulated less TAGs and CrABCA2 overexpression lines accumulated more TAGs than their untransformed parental lines. Transmission electron microscopy showed that CrABCA2 was localized in swollen ER. These results suggest that CrABCA2 transports substrates for TAG biosynthesis to the ER during nitrogen starvation. Our study provides a potential tool for increasing lipid production in microalgae.

Studies on the Use of Carboxymethylcellulose Sodium Salt Matrix for the Immobilization of Photobacterium phosphoreum (Photobacterium phosphoreum을 고정화하기 위한 Carboxymethylcellulose Sodium Salt [CMC] 담체의 이용성 연구)

  • 이용제;정성제;허문석;전억한
    • KSBB Journal
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    • v.15 no.1
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    • pp.49-54
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    • 2000
  • Bioluminescence of Photobacterium phosphoreum has been used for the detection of pollutants in the environment. Immobilization method was used to maintain the stability of bioluminescence of P. phosphoreum. The carboxymethylcellulose was investigated to find out whether it was suitable for the immobilization of P. phosphoreum as a matrix without disturbing the bioluminescence emission. A maintenance of bioluminescence was determined from the P. phosphoreum immobilized on the various concentrations of carboxymethylcellulose. A relatively high bioluminescence intensity was shown with immobilized cells on 1%(w/v) carboxymethylcellulose. The effect of carboxymethylcellulose concentrations on the sensitivity of Crcompounds including $Na_{2}CrO_{4}$, $K_{2}CrO_{4}$, $CrO_{3}$, CrK$(SO_4)_{2}$ and $CrCl_{3}$ to the bioluminescence intensity. The calculated $EC_{50}$ showed that the linear relations between such substances and bioluminesence intensity were established.

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La0.8Ca0.2CrO3 Interconnect Materials for Solid Oxide Fuel Cells: Combustion Synthesis and Reduced-Temperature Sintering

  • Park, Beom-Kyeong;Lee, Jong-Won;Lee, Seung-Bok;Lim, Tak-Hyoung;Park, Seok-Joo;Song, Rak-Hyun;Shin, Dong-Ryul
    • Journal of Electrochemical Science and Technology
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    • v.2 no.1
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    • pp.39-44
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    • 2011
  • Sub-micrometer $La_{0.8}Ca_{0.2}CrO_3$ powders for ceramic interconnects of solid oxide fuel cells were synthesized by the aqueous combustion process. The materials were prepared from the precursor solutions with different glycine (fuel)-to-nitrate (oxidant) ratios (${\phi}$). Single-phase $La_{0.8}Ca_{0.2}CrO_3$ powders with a perovskite structure were obtained after combustion when ${\phi}$ was equal to or larger than 0.480. Especially, the stoichiometric precursor with ${\phi}$ = 0.555 yielded the spherical $La_{0.8}Ca_{0.2}CrO_3$ particles with 150-250 nm diameters after calcination at $1000^{\circ}C$. When compared with the powders synthesized by the solid-state reaction, the combustion-derived, fine powders exhibited improved sinterability, leading to near-full densification at $1400^{\circ}C$ in oxidizing atmospheres. Moreover, a small quantity of glass additives was used to reduce the sintering temperature, and considerable densification was indeed achieved at temperatures as low as $1100^{\circ}C$.

Respones of Photobacterium phosphoreum to toxic substances (독성물질 농도에 따른 Photobacterium phosphoreum의 bioluminescence 변화)

  • 정계훈;정성제;이용제;허문석;전억한
    • Microbiology and Biotechnology Letters
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    • v.28 no.1
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    • pp.45-51
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    • 2000
  • Photobacterium phosphoreum was used for the study of bioluminescence response to toxic substances including phenol, As2O3, SoO2, and CrO3 in view of developing monitoring system. measurement of inhibition of bioluminescence in P. phosphoreum has been proposed as a sensitive and raped procedure to monitor toxic substances. The concentration of toxic substance causing 50% light reduction(EC50) in bioluminescence intensity was determined with free and immobilized P. phosphoreum, The minimum inhibitory concentrations (MICs) for bioluminescence emission were found to be 400ppm for As2O3, 800ppm for phenol, 60ppm for SeO2 and 60ppm for CrO3 , respectively. The linear correlation between Gamma value and the concentration of toxic substances was obtained and EC50 wa calculated from the linear correlation. The free cells were shown to be more sensitive to toxic substances than cells immobilized on Sr-alginate and Ca-alginate. However, the linear regression curves were derived from the Sr-alginate immobilized cells indicating the immobilization method in s useful tool for monitoring of toxic substances under the more stable condition of bioluminescence.

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Immunopreventive Effects against Murine H22 Hepatocellular Carcinoma in vivo by a DNA Vaccine Targeting a Gastrin-Releasing Peptide

  • Meko'o, Jean Louis Didier;Xing, Yun;Zhang, Huiyong;Lu, Yong;Wu, Jie;Cao, Rongyue
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.20
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    • pp.9039-9043
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    • 2014
  • There is a continuing need for innovative alternative therapies for liver cancer. DNA vaccines for hormone/growth factor immune deprivation represent a feasible and attractive approach for cancer treatment. We reported a preventive effect of a DNA vaccine based on six copies of the B cell epitope GRP18-27 with optimized adjuvants against H22 hepatocarcinoma. Vaccination with pCR3.1-VS-HSP65-TP-GRP6-M2 (vaccine) elicited much higher level of anti-GRP antibodies and proved efficacious in preventing growth of transplanted hepatocarcinoma cells. The tumor size and weight were significantly lower (p<0.05) in the vaccine subgroup than in the control pCR3.1-VS-TP-HSP65-TP-GRP6, pCR3.1-VS-TP-HSP65-TP-M2 or saline subgroups. In addition, significant reduction of tumor-induced angiogenesis associated with intradermal tumors of H22 cells was observed. These potent effects may open ways towards the development of new immunotherapeutic approaches in the treatment of liver cancer.

Anti-cancer Effects of Samultang-Gami on HeLa, HepG2, AGS Cells (사물탕가미방(四物湯加味方)의 항암효과에 대한 실험적 연구)

  • Jung, Jae-Joong;Goo, Sun-Young;Goo, Sun-Young;Sung, Jung-Suk;Kim, Dong-Il
    • The Journal of Korean Obstetrics and Gynecology
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    • v.23 no.3
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    • pp.38-55
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    • 2010
  • Purpose: This study was designed to find out the anti-cancer effects of Samultang-Gami which was composed of Rehmanniae Radix(RR), Angelicae Gigantis Radix(AGR), Cnidii Rhizoma(CR), Paeoniae Radix(PR), Cortex Moutan Radicis(CMR), Hedyotis Diffusa(HD) and Caesalpinia Sappan on HeLa, HepG2 and AGS cells. Methods: Various cancer cell lines including HeLa, HepG2 and AGS cells, were used. In vitro anti-cancer effects were measured by MTT assay using cancer cell lines treated with various concentrations of 70% ethanol extract of Samultang-Gami. Expression of cell cycle arrest mediators including Bax, Bcl-2, p53 and DARP-1 proteins were measured by Western blot analysis. Results: 1. Samultang-Gami decreased the viability of HeLa and HepG cells in a dosedependent manner. 2. AGR, CMR, PR and HD decreased the viability of HeLa, HepG2 and AGS cells. 3. We could observe that the decreased Bax and Bcl-2 expression level and the increased PARP-1 expression level by Samultang-Gami extracts treated in HeLa cells. 4. We could observe that the decreased Bcl-2 expression level and the increased Bax, p53 and PARP-1 expression level by RR extracts treated in HeLa cells. and also could observe that the reduction of the protein level of Bcl-2, p53 and PARP-1 and the increase of the protein level of Bax by PR in HeLa cells. 5. We could observe that the increased p53 expression level, the decreased PARP-1's that and the unchanged Bax and Bcl-2's that by Samultang-Gami extracts treated in HepG2 cells. 6. We could observe that the reduced Bcl-2 expression level by each of RR extracts and PR extracts in HepG2 cells. 7. The treatment of Samultang-Gami in AGS cells didn't have any effect on the expression level of Bax, Bcl-2, p53 and PARP-1. 8. We could observe that the increased p53 and PARP-1 expression level by each of CR, RR and PR extracts in AGS cells. Conclusion: Taken together, we suggest that Samultang-Gami exhibits cytotoxic effects on HeLa, HepG2 and AGS cells, causing apoptosis. The results showed that Samultang-Gami may do so by regulating the expression of specific target molecules that promote efficient apoptotic cell death in a dose-dependent manner.

Blockage of the Immune Complex-triggered Transmembrane Proximity Between Complement Receptor Type 3 and Microfilaments by Staurosporine and Methyl-2,5-dihydroxycinnamate

  • Poo, Ha-Ryoung;Lee, Young-Ik;Todd, Robert F. III;Petty, Howard R.
    • BMB Reports
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    • v.31 no.1
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    • pp.64-69
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    • 1998
  • Recent studies have suggested that integrin (CR3) participates in the signal transduction pathways of certain GPI-anchored phagocytic receptors including $Fc{\gamma}RIIIB$. One consequence of this functional linkage is an inducible association between CR3 and cortical microfilaments that is triggered by $Fc{\gamma}RIIIB$ binding to immobilized immune complexes (IC). That this signaling event requires the co-expression of $Fc{\gamma}RIIIB$ with CR3 was documented by the use of NIH 3T3 transfectants expressing both CR3 and $Fc{\gamma}RIIIB$ (clone 3-23), CR3 alone (clone 3-19), and $Fc{\gamma}RIIIB$ alone (clone 3-15). Pretreatment of 3-23 cells with protein kinase inhibitors such as staurosporine and methyl 2,5-dihydroxycinnamate (MDHC) blocked IC-stimulated CR3 microfilament proximity without affecting the extent to which $Fc{\gamma}RIIIB$ constrains the lateral membrane mobility of a subset of CR3 on the cell surface (as measured in fluorescence recovery after photobleaching experiments). These data support that CR3 and $Fc{\gamma}RIIIB$ molecules are physically and functionally associated and that ligation of FcgRIIIB triggers CR3-dependent signal transduction.

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Screening for Mucosal Protective Effects of Various Korean Herbal Medicine Extracts in Gastroesophageal Reflux Disease (한방 추출물의 역류성 식도염 점막보호 효과에 대한 스크리닝)

  • Il-ha Jeong;Min Ju Kim;Mi-Rae Shin;Seong-Soo Roh
    • The Korea Journal of Herbology
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    • v.39 no.1
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    • pp.39-47
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    • 2024
  • Objectives : This study evaluates how various traditional Korean herbal medicines assess MUC5AC expression for esophageal mucosal defense and analyzes the associated mechanisms involved in inflammation. Methods : Forty types of traditional Korean herbal medicines were assessed for in vitro antioxidant activities, and the real-time PCR method was employed to analyze MUC5AC expression under pH 4.5 conditions in human esophageal epithelial cells (HET-1A). Eight types of Korean herbal medicines were evaluated for in vitro antioxidant activities, and Reactive oxygen specise (ROS) expression was analyzed under bile salt (480 𝜇M) and pH 5.5 conditions in human esophageal epithelial cells (HET-1A). Simulation experiments involving bile salts and acidity were conducted for one hour to assess the efficacy of four drugs, and the activities of Mitogen-activated Protein Kinase (MEK), Nuclear Factor Kappa B (NF-𝜅B), and Cyclooxygenase-2 (COX-2) were detected through Western blot analysis. Results : Compared to the Normal group, the Control group exhibited higher ROS generation. Such increased ROS levels were significantly reduced by four extracts: Citrus Unshius Pericarpium (CUP), Cnidium officinale Rhizoma (CR), Ginseng Radix (GR), and Linderae Radix (LR). The protein expression of COX-2 decreased with the treatment of LR, CUP, and CR. Particularly, CUP and CR exhibited superior effects compared to other groups in inhibiting the phosphorylation of NF-𝜅B. Conclusion : Based on the results obtained, we have identified drugs that inhibit oxidative stress and inflammation caused by bile acid in esophageal epithelial cells. Our future plans involve comparing and analyzing the efficacy of these herbal medicines through animal experiments.