• Title/Summary/Keyword: C2C12 cell

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Anti-fatigue activity of a mixture of seahorse (Hippocampus abdominalis) hydrolysate and red ginseng

  • Kang, Nalae;Kim, Seo-Young;Rho, Sum;Ko, Ju-Young;Jeon, You-Jin
    • Fisheries and Aquatic Sciences
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    • v.20 no.3
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    • pp.3.1-3.8
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    • 2017
  • Seahorse, a syngnathidae fish, is one of the important organisms used in Chinese traditional medicine. Hippocampus abdominalis, a seahorse species successfully cultured in Korea, was validated for use in food by the Ministry of Food and Drug Safety in February 2016; however. the validation was restricted to 50% of the entire composition. Therefore, to use H. abdominalis as a food ingredient, H. abdominalis has to be prepared as a mixture by adding other materials. In this study, the effect of H. abdominalis on muscles was investigated to scientifically verify its potential bioactivity. In addition, the anti-fatigue activity of a mixture comprising H. abdominalis and red ginseng (RG) was evaluated to commercially utilize H. abdominalis in food industry. H. abdominalis was hydrolyzed using Alcalase, a protease, and the effect of H. abdominalis hydrolysate (HH) on the muscles was assessed in C2C12 myoblasts by measuring cell proliferation and glycogen content. In addition, the mixtures comprising HH and RG were prepared at different percentages of RG to HH (20, 30, 40, 50, 60, 70, and 80% RG), and the anti-fatigue activity of these mixtures against oxidative stress was assessed in C2C12 myoblasts. In C2C12 myoblasts, $H_2O_2$-induced oxidative stress caused a decrease in viability and physical fatigue-related biomarkers such as glycogen and ATP contents. However, treatment with RG and HH mixtures increased cell viability and the content of fatigue-related biomarkers. In particular, the 80% RG mixture showed an optimum effect on cell viability and ATP synthesis activity. In this study, all results indicated that HH had anti-fatigue activity at concentrations approved for use in food by the law in Korea. Especially, an 80% RG to HH mixture can be used in food for ameliorating fatigue.

Induction of Apoptotic Cell Death and Depression of Bcl-2 Protein Levels by Trans-10,cis-12 Conjugated Linoleic Acid in Human Prostate Cancer (인간 전립선 암세포인 TSU-Pr1에서 trans-10,cis-12 Conjugated Linoleic Acid에 의한 Apoptosis 유발과 Bcl-2 단백질의 발현억제)

  • 오윤신;김은지;이상곤;정차권;강일준;신현경;윤정한
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.6
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    • pp.1126-1133
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    • 2002
  • Conjugated linoleic acid (CLA) is a collective term for a class of positional and geometric conjugated dienoic isomers of linoleic acid (LA) and has anti-cancer activity in experimental animals. We have previously observed that an isomeric mixture of CLA and trans-10,cis-12 (t10c12) inhibited cell growth in a dose-dependent manner whereas LA and cis-9,trans-11 (c9t11) had no effect. The present study examined whether the CLA mixture and t10c12 induce apoptotic cell death. TSU-Prl cells were incubated for three days in serum-free medium in the absence or presence of individual fatty acids, and the DNA fragmentation assay was performed. Cells treated with the CLA mixture or t10c12 produced a distinct oligonucleosomal ladder with different sizes of DNA fragments, a typical characteristic of cells undergoing apoptosis. By contrast, LA and c9t11 had no effect. Western immunoblot analysis of total lysates revealed that t10c12 reduced anti-apoptotic, 26 kDa, Bcl-2 protein levels by 49$\pm$8% compared with controls, whereas this CLA isomer did not alter pro-apoptotic,21 kDa, Bax protein levels. These results suggest that growth inhibitory effect of the t10c12 CLA isomer may, at least in part, be attributed to Increased apoptotic death in TSU-Prl cells.

Potential role of exercise-induced glucose-6-phosphate isomerase in skeletal muscle function

  • Kwak, Seong Eun;Shin, Hyung Eun;Zhang, Di Di;Lee, Jihyun;Yoon, Kyung Jin;Bae, Jun Hyun;Moon, Hyo Youl;Song, Wook
    • Korean Journal of Exercise Nutrition
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    • v.23 no.2
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    • pp.28-33
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    • 2019
  • [Purpose] Recent studies have shown that glucose-6-phosphate isomerase (GPI)-which is a glycolysis interconversion enzyme-reduces oxidative stress. However, these studies are limited to tumors such as fibrosarcoma, and there are no studies that have examined the effects of exercise on GPI expression in mice skeletal muscle. Furthermore, GPI acts in an autocrine manner thorough its receptor, autocrine motility factor receptor (AMFR); therefore, we investigated expression level changes of secreted GPI from skeletal muscle in in vitro study to examine the potential role of GPI on skeletal muscle. [Methods] First, we performed an in vitro study, to identify the condition that upregulates GPI levels in skeletal muscle cells; we treated C2C12 muscle cells with an exercise-mimicking chemical, AICAR. AICAR treatment upregulated GPI expression level in C2C12 cell and its secretomes. To confirm the direct effect of GPI on skeletal muscle cells, we treated C2C12 cells with GPI recombinant protein. [Results] We found that GPI improved the viability of C2C12 cells. In the in vivo study, the exercise-treated mice group showed upregulated GPI expression in skeletal muscle. Based on the in vitro study results, we speculated that expression level of GPI in skeletal muscle might be associated with muscle function. We analyzed the association between GPI expression level and the grip strength of the all mice group. The mice group's grip strengths were upregulated after 2 weeks of treadmill exercise, and GPI expression level positively correlated with the grip strength. [Conclusion] These results suggested that the exercise-induced GPI expression in skeletal muscle might have a positive effect on skeletal muscle function.

Expression and cDNA Cloning of klp-12 Gene Encoding an Ortholog of the Chicken Chromokinesin, Mediating Chromosome Segregation in Caenorhabditis elegans

  • Ali, M. Yusuf;Khan, M.L.A.;Shakir, M.A.;Kobayashi, K. Fukami;Nishikawa, Ken;Siddiqui, Shahid S.
    • BMB Reports
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    • v.33 no.2
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    • pp.138-146
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    • 2000
  • In eukaryotes, chromosomes undergo a series of complex and coordinated movements during cell division. The kinesin motor proteins, such as the chicken Chromokinesin, are known to bind DNA and transport chromosomes on spindle microtubles. We previously cloned a family of retrograde C-terminus kinesins in Caenorhabditis elegans that mediate chromosomal movement during embryonic development. Here we report the cloning of a C. elegans klp-12 cDNA, encoding an ortholog of chicken Chromokinesin and mouse KIF4. The KLP-12 protein contains 1609 amino acid and harbors two leucine zipper motifs. The insitu RNA hybridization in embryonic stages shows that the klp-12 gene is expressed during the entire embryonic development. The RNA interference assay reveals that, similar to the role of Chromokinesin, klp-12 functions in chromosome segregation. These results support the notion that during mitosis both types, the anterograde N-terminus kinesins such as KLP-12 and the retrograde C-terminus kinesins, such as KLP-3, KLP-15, KLP-16, and KLP-17, may coordinate chromosome assembly at the metaphase plate and chromosomal segregation towards the spindle poles in C. elegans.

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Purification and characterization of biochemical properties of hemolysin from Vibrio fluvialis (Vibrio fluvialis 유래의 hemolysin 정제와 생화학적 특성)

  • 이종희;한정현;안선희;김선회;이은미;공인수
    • Journal of Life Science
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    • v.12 no.4
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    • pp.490-495
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    • 2002
  • Hemolysin (VFH) of V. fluvialis, which is a pathogenic bacteria, causing watery diarrhea with vomiting, abdominal croup, was purified. V. fluvialis was cultivated in BHI medium and the culture supematant was precipitated by ammonium sulfate. The protein was purified by chromatographies on columns of DEAE-cellulose and Mono-Q. Molecular weight of the purified VFH was estimated as 79kDa by SDS-PAGE. The optimal temperature for a maximum hemolytic activity was at around 35$^{\circ}C$ and the activity was decreased at 4$0^{\circ}C$ Cytotoxicity of VFH was also investigated using RTG-2 cell line. LDH assay study showed that 50$\mu\textrm{g}$/m1 of VFH release 80% of total cellular LDH (lactate dehydrogenase) from RTG-2 cell and microscopic observation also showed the morphological change of cell.

The Effect of Conjugated Linoleic Acid and Its Isomers on the Proliferation of Prostate TSU-Prl Cancer Cells (Conjugated Linoleic Acid (CLA)와 그 이성체가 전립선 암세포의 증식에 미치는 영향)

  • 오윤신;김은지;김종우;김우경;이현숙;윤정한
    • Journal of Nutrition and Health
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    • v.35 no.2
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    • pp.192-200
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    • 2002
  • Conjugated linoleic acid (CLA) is a collective term for positional and geometric isomers of octadecadienoic acid in which the double bonds are conjugated. CLA has anticancer activity in a variety of animal cancer models, and cis-9, trans-11 (c9t11) and trans-10, cis-12(t10c12) CLA are the most predominant isomers present in the synthetic preparations utilized in these animal studies. To compare the ability of c9t11, t10c12 and an isomeric mixture of CLA to inhibit TSU-Prl cell growth, cells were incubated in a serum-free medium with various concentrations of these fatty acids. The isomeric mixture inhibited cell growth in a dose-dependent manner (1-3 $\mu$M) with a 41 $\pm$ 1% inhibition observed at 3 $\mu$M concentration after 48 hours. T10c12 also inhibited cell proliferation in a dote-dependent manner, However, the efficacy and potency of this isomer was much greater than that of the isomeric mixture with a 49 $\pm$ 2% inhibition observed at 0.3 $\mu$M concentration after 48 hours. By contrast, c9t11 slightly increased cell proliferation. To determine whether the growth-inhibiting effect of CLA is related to the changes in production of insulin-like growth factors (IGF) and IGF-binding proteins (IGFBP) by these cells, serum-free conditioned media were collected. Immunoblot analysis of conditioned media using a monoclonal anti-IGF-II antibody showed that both the isomeric mixture and t10c12 inhibited secretion of both mature 7,500 Mr and higher Mr forms of pro IGF-II, whereas c9t11 had no effect. Ligand blot analysis with 125I-IGF-II revealed the presence of two types of IGFBPs : 24,000 Mr IGFBP-4 and 30,000 Mr IGFBP-6. The production of IGFBP-4 slightly decreased at the highest concentrations of the isomeric mixture and t10c12. These results indicate that CLA inhibits human prostate cancer cell growth, an effect largely due to the action of t10c12. The growth inhibition may result, at least in part, from decreased production of IGF-II and IGFBP-4 by these cells.

Factors Influencing Siderophore Production by Plant Growth Promoting Rhizopseudomonas Strains (식물생장촉진 근권 Pseudomonas의 Siderophore 생성에 관여하는 요인)

  • Seong, Ki-Young
    • Korean Journal of Soil Science and Fertilizer
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    • v.28 no.3
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    • pp.287-294
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    • 1995
  • This study demonstrates the influence of environmental conditions, other than iron, on pyoverdin production by fluorescent Pseudomosonas. In slightly acidic pH conditions(pH 6), cell yield was reduced while the siderophore production per cell yield was increased. The optimum temperatures for the siderophore production and cell yield was $19^{\circ}C$ and $28^{\circ}C$ for 7NSK2 and $12^{\circ}C$ and $19^{\circ}C$ for ANP15. The carbon and nitrogen balance showed that at low C : N ratio of the growth medium (higher nitrogen concentration), both cell yield and siderophore production was reduced. Use of different carbon sources revealed that citrate as a carbon source facilitated iron uptake and resulted in a significant reduction in siderophore production. However, at the late exponential phase, the iron content in the cell biomass was not significantly different from those grown in glucose or succinate. From these results it can be suggested that the environmental factors other than iron may also influence siderophore production by fluorescent pseudomonas.

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Propofol Post-conditioning Protects against COS-7 Cells in Hypoxia/reoxygenation Injury by Induction of Intracellular Autophagy

  • Kwak, Jin-Won;Kim, Eok-Nyun;Park, Bong-Soo;Kim, Yong-Ho;Kim, Yong-Deok;Yoon, Ji-Uk;Kim, Cheul-Hong;Yoon, Ji-Young
    • Journal of The Korean Dental Society of Anesthesiology
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    • v.14 no.1
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    • pp.49-56
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    • 2014
  • Background: Propofol (2.6-diisopropylphenol) is a widely used intravenous anesthetic agent for the induction and maintenance of anesthesia during surgeries and sedation for ICU patients. Propofol has a structural similarity to the endogenous antioxidant vitamin E and exhibits antioxidant activities.13) However, the mechanism of propofol on hypoxia/reoxygenation (H/R) injury has yet to be fully elucidated. We investigated how P-PostC influences the autophagy and cell death, a cellular damage occurring during the H/R injury. Methods: The groups were randomly divided into the following groups: Control: cells were incubated in normoxia (5% CO2, 21% O2, and 74% N2) without propofol treatment. H/R: cells were exposed to 24 h of hypoxia (5% CO2, 1% O2, and 94% N2) followed by 12 h of reoxygenation (5% CO2, 21% O2, and 74% N2). H/R + P-PostC: cells post-treated with propofol were exposed to 24 h of hypoxia followed by 12 h of reoxygenation. 3-MA + P-PostC: cells pretreated with 3-MA and post-treated propofol were exposed to 24 h of hypoxia followed by 12 h of reoxygenation Results: The results of our present study provides a new direction of research on mechanisms of propofol-mediated cytoprotection. There are three principal findings of these studies. First, the application of P-PostC at the onset of reoxygenation after hypoxia significantly increased COS-7 cell viability. Second, the cellular protective effect of P-PostC in H/R induced COS-7 cells was probably related to activation of intra-cellular autophagy. And third, the autophagy pathway inhibitor 3-MA blocked the protective effect of P-PostC on cell viability, suggesting a key role of autophagy in cellular protective effect of P-PostC. Conclusions: These data provided evidence that P-PostC reduced cell death in H/R model of COS-7 cells, which was in agreement with the protection by P-PostC demonstrated in isolated COS-7 cells exposed to H/R injury. Although the this study could not represent the protection by P-PostC in vivo, the data demonstrate another model in which endogenous mechanisms evoked by P-PostC protected the COS-7 cells exposed to H/R injury from cell death.

Immuno-stimulating and anti-metastatic activities of the polysaccharides isolated from Angelica gigas (참당귀로부터 분리한 다당의 면역증진 활성과 항전이 활성)

  • Son, Seung-U;Shin, Kwang-Soon
    • Korean Journal of Food Science and Technology
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    • v.53 no.3
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    • pp.304-312
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    • 2021
  • The present study aimed to develop new physiologically active ingredients from Angelica gigas. The polysaccharides purified from A. gigas, AGE-2c-I, showed potent anti-complementary activity in a dose-dependent manner. C3 activation products were identified through crossed immuno-electrophoresis using anti-human C3 antibodies and the anti-complementary activity of AGE-2c-I under Ca++-free conditions suggests that AGE-2c-I may induce complementary activation via both alternative and classical pathways. In addition, AGE-2c-I augmented the production of various cytokines, such as interleukin (IL)-6, IL-10, IL-12, and tumor necrosis factor-α, by peritoneal macrophages. Furthermore, intravenous (i.v.) administration of AGE-2c-I dose-dependently enhanced natural killer cell cytotoxicity against YAC-1 lymphoma. In experimental lung metastasis, prophylactic i.v. administration of AGE-2c-I inhibited lung metastasis by 58% at 100 ㎍/mouse. From the above results, we suggest that AGE-2c-I purified from A. gigas has potent immune system-stimulating activities, and is a potentially promising food ingredient beneficial to human health.

High-Voltage Liquid-Electrolyte Microbatteries Inspired from Electric Eels (전기뱀장어의 전기발생을 모사한 고전압 액체 전해질 미소전지)

  • Kim, Mun-Chul;Cho, Young-Ho
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.33 no.5
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    • pp.469-473
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    • 2009
  • We present high-voltage liquid-electrolyte microbatteries, inspired from the high-voltage generation mechanism of electric eels using serially connected multiple-cell arrays. In the microbatteries, we purge air into the electrolyte filled in a channel layer to isolate serially connected multiple cell arrays using three surface-tension valves (cell-front, outlet, and cell-end valves). Compared to the previous multi-cell stack or interconnection, present microbatteries provide a reduced multi-cell charging time. We have designed and characterized four different prototypes C1, C10, C20, and C40 having 1, 10, 20, and 40 cells, respectively. In the experimental study, the threshold pressures of cell-front, outlet, and cell-end valves were measured as $460{\pm}47$, $1,000{\pm}53$, and $2,800{\pm}170$ Pa, respectively. The average charging time for C40 was measured as $26.8{\pm}4.9$ seconds where the electrolyte and air flow-rates are 100 and $10{\mu}l/min$, respectively. Microbatteries showed the maximum voltage of 12 V (C40), the maximum power density of $110{\mu}W/cm^2$ (C40), and the maximum power capacity of $2.1{\mu}Ah/cm^2$ (C40). We also proposed a tapered-channel to remove the reaction gas from the cell chamber using a surface tension effect. The present microbatteries are applicable to high-voltage portable power devices.