• Title/Summary/Keyword: Protein bodies

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Induction of Apaopotis by Water Extract of Cordyceps militaris (WECM) in Human Hepatocellular Carcinoma HepG2 Cells. (동충하초 열수 추출물에 의한 인체 간암세포 성장억제 및 apoptosis 유발에 관한 연구)

  • Kim, Kyung-Mi;Park, Cheol;Choi, Yung-Hyun;Lee, Won-Ho
    • Journal of Life Science
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    • v.18 no.6
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    • pp.804-813
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    • 2008
  • Cordyceps militaris, the Chinese medicinal fungal genus Cordyceps, is reported to possess many pharmacological activities including immunological stimulating, anti-cancer, anti-virus and anti-infection activities. However, the molecular mechanisms of C. militaris on biochemical actions in cancer have not been clearly elucidated yet. In the present study, we investigated the anti-proliferative activity of the water extract of C. militaris (WECM) in human hepatocellular carcinoma HepG2 cells. It was found that WECM could inhibit the cell growth in a dose-dependent manner, which was associated with morphological changes and apoptotic cell death such as formation of apoptotic bodies and increased populations of apoptotic sub-G1 phase. Apoptotic cell death of HepG2 cells by WECM was connected with a up-regulation of pro-apoptotic Bax expression, tumor suppressor p53 and cyclin-dependent kinase inhibitor p21 (WAF1/CIP1). In addition, WECM treatment induced the proteolytic activation of caspase-3 and a concomitant degradation and/or inhibition of poly (ADP-ribose) polymerase (PARP), ${\beta}-catenin$ and phospholipase $(PLC)-{\gamma}1$ protein. Furthermore, caspase-3 inhibitor, z-DEVD-fmk, significantly inhibited WECM-induced apoptosis demonstrating the important role of caspase-3 in the observed cytotoxic effect. Taken together, these findings provide important new insights into the possible molecular mechanisms of the anti-cancer activity of C. militaris.

Isolation of Anticarcinogenic Isoflavone-conjugated Glycoproteins from a Submerged Liquid Culture of Agaricus blazei Mycelia by the Autolysis Process (신령버섯균사체 액체배양물의 자가분해에 의한 항암성 isoflavone-conjugated glycoprotein 분리)

  • Kim, So Young;Kim, Young Suk;Jang, Joung Soon;Kim, Boh Hyun;Rakib, Abdur Md.;Kim, Gon Sup;Kim, Jeong Ok;Ha, Yeong Lae
    • Journal of Life Science
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    • v.24 no.12
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    • pp.1316-1324
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    • 2014
  • Most beta-glucans obtained from various fruit bodies of mushrooms and mushroom mycelial cultures have high-molecular weight glycoproteins, conjugated with beta-glucans. We report that isoflavone-conjugated glycolproteins (designated as gluvone) were isolated and exhibited stronger anticarcinogenic activities. Agaricus blazei mycelia (ABM) was cultured in a liquid medium containing soybean flakes for 14 days. The liquid culture was autolyzed by incubating at $53^{\circ}C$ (pH 5.5) for 3 h. A crude glycoprotein (CGP) fraction with a cytotoxic effect on a mouse ascite cancer cell line (S-180) and a human breast cancer cell line (MCF-7) was isolated from the autolyzed ABM cultures by 80% ethanol treatment. Gluvone was isolated from the CGP with Sephadex G-75 column chromatography. It exhibited a stronger anticancer effect than CGP against the S-180 cell-induced female ICR mouse ascites carcinogenesis. Gluvone with 9,400 daltons was identified as a glycoprotein conjugated with isoflavone. According to HPLC and GC analysis, in conjunction with $^1H$-NMR spectral analysis, it contained 60% carbohydrates (glucose, fructose, and ribose), 31% protein, and 2% isoflavone (daidzein and genistein), which is a novel material. These results indicate that a strong anticarcinogenic gluvone was isolated from the autolyzed product of a submerged liquid culture of ABM, suggesting that autolysis could be a useful tool to produce antitumor agents.

Anti-cancer effect of farrerol induced apoptosis through activating p38 MAPK in Human breast cancer MCF-7 cells (인간 유방암 세포주 MCF-7에 대한 farrerol의 p38 MAPK 활성화와 세포사멸 유도를 통한 항암 효과)

  • Chae, Jongbeom;Lee, Seul Gi;Nam, Ju-Ock
    • Journal of Applied Biological Chemistry
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    • v.63 no.2
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    • pp.147-152
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    • 2020
  • Farrerol is a flavanone isolated from the traditional Chinese herb 'Man-shan-hong' (Rhododendron dauricum L.). Farrerol has been reported to have various bioactivities including anti-oxidant, anti-inflammation, and anti-fungal. However, anti-cancer effect of farrerol has not yet been reported in MCF-7 breast cancer cells. In the present study, we investigated the anti-cancer effect of farrerol on MCF-7 cells. Farrerol decreased viability and induced apoptosis of MCF-7 cells in a dose dependent manner. Ferrerol exhibited a significant anti-proliferation effect with a half-maximal inhibitory concentration (IC50) values of 145.04±1.4 μM in MTT assay, when MCF-7 cells were treated with ferrerol for 48 h. Also, ferrerol induced apoptotic bodies of MCF-7 cells as evaluated by TUNEL assay and Annexin V/PI staining using FACS. By mechanism of action, ferrerol regulated the activation of p38 mitogen-activated protein kinase and altered the expression level of BAX, Bcl-2, and Poly ADP Ribose Polymerase in MCF-7 cells. In summary, our finding demonstrated that ferrerol has anti-cancer effect through regulating the activation and expression of apoptosis-related proteins in MCF-7 cells.

Development of Dietary Schedule for Improved Nutritional Support during Training Period of Junior & Senior High School Fencers in Ulsan City (울산지역 중.고등학교 펜싱선수의 적극적인 영양지원을 위한 식사 일정 개발)

  • Cho, Seong-Suk;Lee, Han-Jun
    • Journal of the Korean Dietetic Association
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    • v.17 no.1
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    • pp.32-46
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    • 2011
  • This study was conducted in order to improve nutritional support based on analysis of the eating habits and training schedule of junior and senior high school fencers. A survey was conducted on 30 fencers (junior boys=9, girls=7; senior boys=4, girls=10) using a questionnaire. Total daily calorie intakes were 2325.7${\pm}$1168.5 kcal (boy), 2344.0${\pm}$786.3 kcal (girl) for junior fencers and 2183.3${\pm}$726.6 kcal (boys), 2654.4${\pm}$1043.6 kcal (girls) for senior fencers. Nutrients below the RDIs were riboflavin and vitamin C for junior boy fencers and vitamin A, riboflavin, niacin, and vitamin C for senior boy fencers. The rates of the participants who had obtained nutritional information were 44.4% (boys) and 14.3% (girls) for junior fencers and 50.0% (boys) and 80.0% (girls) for senior fencers. Approximately half of the fencers had meals twice a day (55.6% for junior boy fencers, 57.1% for junior girl fencers, and 50.0% for senior fencers). The number of the fencers who were interested in nutrition varied: 66.7% and 28.6% of the junior boy and girl fencers answered they were interested in nutrition, while 25% and 60% of the senior boy and girl fencers showed interest in nutrition issues. Recommendations of total calories were 2,874 kcal (boys) and 2,377 kcal (girls) for junior fencers and 3,398 kcal (boys) and 2,375 kcal (girls) for senior fencers respectively. A dietary plan for the fencing players during training periods was designed to improve their health and performance. Athletes can simultaneously refuel and repair their bodies while contributing to their rehydration goals by consuming fluids that are sources of carbohydrates and protein, e.g., flavored milk and liquid meal supplement. Specific sport nutrition education applicable to athletes, especially young athletes, is recommended under professional support. Professionals may then target particular categories of athletes for the development of educational programs aimed at improving growth and athletic performance.

The Effects of Tofu (Soybean curd) Containing Mushrooms on the Immune Activities (버섯첨가 두부의 면역활성에 미치는 영향)

  • Kim, Jong-Bong;Lee, Yong-Ho
    • Food Science and Preservation
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    • v.18 no.6
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    • pp.961-966
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    • 2011
  • This research was carried out to evaluate the value of tofu containing mushroom as a immunomodulator. Tofu was made using $CaSO_4{\cdot}2H_2O$ or Lactobacillus extract as a coagulant after adding powder of fruit bodies or mycelia of Letino edodes and Lepista nuda to soybean milk. Proximate compositions of tofu and tofu containing mushroom were analyzed. Levels of interferon ${\gamma}$ (IFN-${\gamma}$), interleukin 4 (IL-4) and tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) in culture media of lymphocytes collected from mouse spleens after being injected with mushroom, regular tofu, or tofu made with mushroom were measured by sandwich ELISA. In addition, concentrations of IgG1, IgG2a and IgE in plasma or lymphocyte culture media were analyzed. Crude protein, crude lipid and crude ash were decreased in tofu containing mushroom but phosphorus was increased significantly. IFN-${\gamma}$ concentration was significantly decreased in mice injected with fruit body or tofu alone. IL-4 level was decreased significantly in mice injected with tofu containing fruit body of L. edodes. However, TNF-${\alpha}$ was increased in mice injected with tofu containing fruit body of L. edodes. Plasma levels of IgG1 were increased in almost all groups, while there was no significant change in IgG2a levels among treated mice groups. Concentrations of IgG1 and IgG2a were increased significantly in lymphocyte culture media of mice injected with tofu containing mushroom. Plasma levels of IgE level was significantly increased in mice injected with tofu or fruit body of L. edodes, but not in mice treated with tofu containing mushroom. These results showed that tofu with mushroom affected immune activities, and it seems valuable to consider developing the mixture of tofu and L. edodes as an immunomodulator.

Induction of apoptosis by water extract Glycyrrhizae radix in human bladder T24 cancer cells (인체 방광암 T24 세포에서 Glycyrrhizae radix 열수추출물에 의한 apoptosis 유도)

  • Eom, Jung Hye;Hwang, Buyng Su;Jeong, Yong Tae;Kim, Min-Jin;Shin, Su Young;Kim, Chul Hwan;Lee, Seung Young;Choi, Kyung Min;Cho, Pyo Yun;Jeong, Jin-Woo;Oh, Young Taek
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.04a
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    • pp.111-111
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    • 2019
  • Glycyrrhizae radix is one of the most frequently prescribed ingredients in Oriental medicine, and G. radix extract has been shown to exert anti-cancer effects. However, the cellular and molecular mechanisms of apoptosis by G. radix are poorly defined. In the present study, it was examined the biochemical mechanisms of apoptosis by water extract of G. radix (WEGR) in human bladder T24 cancer cells. It was found that WEGR could inhibit the cell growth of T24 cells in a dose-dependent manner, which was associated with the induction of apoptotic cell death, as evidenced by the formation of apoptotic bodies, DNA fragmentation and increased populations of annexin-V positive cells. The induction of apoptotic cell death by WEGR was connected with an up-regulation of pro-apoptotic Bax protein expression and down-regulation of anti-apoptotic Bcl-2 and Bcl-xL proteins, and inhibition of apoptosis family proteins (XIAP, cIAP-1 and cIAP-2). In addition, apoptosis-inducing concentrations of WEGR induced the activation of caspase-9, an initiator caspase of the mitochondrial-mediated intrinsic pathway, and caspase-3, accompanied by proteolytic degradation of poly (ADP-ribose)-polymerase. WEGR also induced apoptosis via a death receptor-mediated extrinsic pathway by caspase-8 activation, resulting in the down-regulation of total Bid and suggesting the existence of cross-talk between the extrinsic and intrinsic pathways. Taken together, the present results suggest that WEGR may be a potential chemotherapeutic agent for the control of human bladder cancer cells.

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Sawdust Substitution in Growth Medium of Oyster Mushroom for Using Its By-product Spent Mushroom Substrates as Ruminant Feed (수확 후 배지의 가축 사료화를 위한 느타리 생육배지 톱밥 대체재료 선발 연구)

  • Kim, Jeong-Han;Jang, Myoung-Jun
    • The Korean Journal of Mycology
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    • v.48 no.4
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    • pp.407-414
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    • 2020
  • To replace the sawdust in the growth medium of oyster mushroom to utilize its by-product spent mushroom substrates (SMS) as feed for ruminant, we performed cultivation test using cotton seed hull pellet (CSHP), corn stalk pellet (CSP), corncob (CC), and analyzed the feed chemical properties of those SMS. As a result of cultivation test, CC and CSHP treatment took 27 days for spawn run, 4 days for primordium formation, and 3 days for development fruiting bodies, resulting in a total cultivation period of 34 days. The yield per bottle was 134 g for CC treatment, similar to 130 g for control, while CSHP treatment (112 g) and CSP treatment (68 g) were lower than that of control. The highest biological efficiency (BE) was shown in CC treatment as 80.1%, which was 11.4% higher than 68.7% of control. The SMS of CC treatment had a relatively low content of neutral detergent fiber and acid detergent fiber, and in particular, lignin content was the lowest and crude protein content was the highest among other treatments. Therefore, CC as a substitute material for sawdust was capable of stable mushroom production and excellent nutritional value as a feed for its by-products.

Protective effects of Atractylodis Rhizoma Alba Extract on seizures mice model (뇌전증 동물 모델에 대한 백출 추출물의 보호 효과)

  • Kang, Sohi;Lee, Su Eun;Lee, Ayeong;Seo, Yun-Soo;Moon, Changjong;Kim, Sung Ho;Lee, Jihye;Kim, Joong Sun
    • The Korea Journal of Herbology
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    • v.36 no.6
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    • pp.1-8
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    • 2021
  • Objectives : Atractylodis rhizoma Alba has been traditionally used as a medicinal resource that is used for enhancing Qi (氣) in traditional medicine in Korea, China, and Japan. This study investigated the protective effects of Atractylodis rhizoma Alba extract (ARE) against trimethyltin (TMT), a neurotoxin that causes selective hippocampal injury, using both in vitro and in vivo models. Methods : We investigated the effects of ARE on TMT- (5mM) induced cytotoxicity in primary cultures of mouse hippocampal cells (7 days in vitro ) and on hippocampal injury in C57BL/6 mice injected with TMT (2.6 mg/kg). Results : We observed that ARE treatment (0 - 50 ㎍/mL) significantly reduced TMT-induced cytotoxicity in cultured hippocampal neurons in a dose-dependent manner, based on results of lactate dehydrogenase and 3-4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide assays. Additionally, this study showed that orally administered ARE (5 mg/kg; between -6 and 0 days before TMT injection) significantly attenuated seizures in adult mice. Furthermore, quantitative analysis of allograft inflammatory factor-1 (Iba-1)- and glial fibrillary acidic protein (GFAP)- positive cells showed significantly reduced levels of Iba-1- and GFAP-positive cell bodies in the dentate gyrus of mice treated with ARE prior to TMT injection. These findings indicate the significant protective effects of ARE against the TMT-induced massive activation of microglia and astrocytes in the hippocampus. Conclusions : We conclude that ARE minimizes the detrimental effects of TMT-induced hippocampal neurotoxicity, both in vitro and in vivo . Our findings may serve as useful guidelines to support ARE administration as a promising pharmacotherapeutic approach to hippocampal degeneration.

The immune enhancement effect of Cheonggukjang Water Extract (CWE) via activation of NF-κB pathways in murine macrophage RAW 264.7 cells (RAW 264.7 대식세포에서 청국장 열수 추출물(Cheonggukjang Water Extract, CWE)의 면역 증강 효과)

  • Sehyeon Jang;San Kim;Se Jeong Kim;Sung Ran Yoon;Bo Ram So;Jung A Ryu;Jeong Min Park;Sung Keun Jung
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.282-288
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    • 2023
  • Due to the COVID-19 pandemic, the immuneenhancing health functional food market that protects our bodies from pathogens such as viruses continues to grow. In this study, we aimed to prove the Cheonggukjang, a high-nutrient food with high protein, fat, and dietary fiber content, as an immuneenhancing nutraceutical. Cheonggukjang water extract (CWE) increased the production of nitric oxide, reactive oxygen species, and cytokines such interleukin (IL)-6, IL-1β, and tumor necrosis factor-α without affecting viability in RAW 264.7 cells. Furthermore, CWE significantly upregulated the expression of inducible nitric oxide synthase and cyclooxygenase-2 in RAW 264.7 cells. CWE enhanced the phosphorylation of I kappa B kinase α/β and I kappa B (IκB)α, as well as the degradation of IκBα. CWE also induced increased phosphorylation of nuclear factor-kappa B p65 and facilitated the redistribution of p65 from the cytoplasm to the nucleus in RAW 264.7 cells. These findings suggest that CWE has potential as a health functional food material that can enhance the innate immune response.

Transduction of eGFP Gene to Human Embryonic Stem Cells and Their Characterization (인간 배아줄기세포로의 eGFP 유전자 도입 및 특성 분석)

  • Kim, Yoon-Young;Ku, Seung-Yup;Park, Yong-Bin;Oh, Sun-Kyung;Moon, Shin-Yong;Choi, Young-Min
    • Clinical and Experimental Reproductive Medicine
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    • v.36 no.4
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    • pp.283-292
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    • 2009
  • Objective: Human embryonic stem cells (hESCs) can proliferate indefinitely and differentiate into all kinds of cell types in vitro. Therefore, hESCs can be used as a cell source for cell-based therapy. Transduction of foreign genes to hESCs could be useful for tracing differentiation processes of hESCs and elucidation of gene function. Thus, we tried to introduce enhanced green fluorescent protein (eGFP) gene to hESCs, XX and XY cell lines in this study. Methods: Lentivirus containing eGFP was packaged in 293T cells and applied to hESCs to transduce eGFP. Expression of transduced eGFP was evaluated under the fluorescence microscope and eGFP positive population was analyzed by FACS. Expression of undifferentiation state markers such as Oct4, Nanog, SSEA4 and Tra-1-81 was examined by RT-PCR and/or immunofluorescence in eGFP-hESCs after transduction. In addition, the ability of eGFP-hESCs to form embryoid bodies (EBs) was tested. Results: eGFP was successfully transduced to hESCs by lentivirus. eGFP expression was stably maintained up to more than 40 passages. eGFP-hESCs retained expression patterns of undifferentiation state markers after transduction. Interestingly, disappearance of transduced eGFP was notably observed during spontaneous differentiation of eGFP-hESCs. Conclusion: We established eGFP expressing hESC lines using lentivirus and showed the maintenance of undifferentiation characteristics of these eGFP-hESCs. This reporter-containing hESCs could be useful for tracing the processes of differentiation of hESCs and other studies.