• Title/Summary/Keyword: culture stress

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Comprehensive proteome analysis using quantitative proteomic technologies

  • Kamal, Abu Hena Mostafa;Choi, Jong-Soon;Cho, Yong-Gu;Kim, Hong-Sig;Song, Beom-Heon;Lee, Chul-Won;Woo, Sun-Hee
    • Journal of Plant Biotechnology
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    • v.37 no.2
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    • pp.196-204
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    • 2010
  • With the completion of genome sequencing of several organisms, attention has been focused to determine the function and functional network of proteins by proteome analysis. The recent techniques of proteomics have been advanced quickly so that the high-throughput and systematic analyses of cellular proteins are enabled in combination with bioinformatics tools. Furthermore, the development of proteomic techniques helps to elucidate the functions of proteins under stress or diseased condition, resulting in the discovery of biomarkers responsible for the biological stimuli. Ultimate goal of proteomics orients toward the entire proteome of life, subcellular localization, biochemical activities, and their regulation. Comprehensive analysis strategies of proteomics can be classified as three categories: (i) protein separation by 2-dimensional gel electrophoresis (2-DE) or liquid chromatography (LC), (ii) protein identification by either Edman sequencing or mass spectrometry (MS), and (iii) quanitation of proteome. Currently MS-based proteomics turns shiftly from qualitative proteome analysis by 2-DE or 2D-LC coupled with off-line matrix assisted laser desorption ionization (MALDI) and on-line electrospray ionization (ESI) MS, respectively, to quantitative proteome analysis. Some new techniques which include top-down mass spectrometry and tandem affinity purification have emerged. The in vitro quantitative proteomic techniques include differential gel electrophoresis with fluorescence dyes, protein-labeling tagging with isotope-coded affinity tag, and peptide-labeling tagging with isobaric tags for relative and absolute quantitation. In addition, stable isotope labeled amino acid can be in vivo labeled into live culture cells through metabolic incorporation. MS-based proteomics extends to detect the phosphopeptide mapping of biologically crucial protein known as one of post-translational modification. These complementary proteomic techniques contribute to not only the understanding of basic biological function but also the application to the applied sciences for industry.

The venom of jellyfish, Chrysaora pacifica, induces neurotoxicity via activating Ca2+-mediated ROS signaling in HT-22 cells

  • Yang, Yoon-Sil;Kang, Young-Joon;Kim, Hye-Ji;Kim, Min-Soo;Jung, Sung-Cherl
    • Journal of Applied Biological Chemistry
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    • v.62 no.4
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    • pp.347-353
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    • 2019
  • Stings of jellyfish, which frequently occur in a warm season, cause severe pain, inflammation and sometimes irreversible results such as the death. Harmful venoms from jellyfish, therefore, have been studied for finding the therapeutic agents to relieve pain or to neutralize toxic components. However, it is still unclear if and how jellyfish venom reveal neuronal toxicity even though pain induction seems to result from the activation of nociceptors such as nerve endings. In this study, using HT-22 cell line, we investigated neurotoxic effects of the venom of Chrysaora pacifica (CpV) which appears in South-East ocean of Korea. In 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, CpV significantly reduced the viability of HT-22 cells in a dose-dependent manner. Additionally, in 2',7'-Dichlorofluorescin diacetate fluorescence test under the culture condition lacking dominant inflammatory factors, CpV remarkably increased the production of intracellular reactive oxygen species (ROS). Reduced responsive fluorescence to Rhodamine123 and increased expression of intracellular cytochrome c were also observed in HT-22 cells treated with CpV. These indicate that CpV-reduced viability of HT-22 cells may be due to the activation of apoptotic signalings mediated with oxidative stress and mitochondrial dysfunction. Furthermore, removing Ca2+ ion or adding N-acetyl-Lcystein remarkably blocked the CpV effect to reduce the viability of HT-22 cells. The findings in this study clearly demonstrate that CpV may activate Ca2+-mediated ROS signalings and mitochondrial dysfunction resulting in neuronal damage or death, and suggest that blocking Ca2+ pathway is a therapeutic approach to possibly block toxic effects of jellyfish venoms.

A Case Report on a Hwabyung Patient with Binge Eating (폭식을 주소로 한 화병(火病)환자 치험(治驗)1예(例))

  • An, Tae-Han;Jang, Jeong-A;Park, Eun-Young;Jeong, Seong-Sik;Kim, Jin-Won;Seo, Ho-Seok;Kim, Jin-Yi
    • The Journal of Korean Medicine
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    • v.32 no.5
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    • pp.139-146
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    • 2011
  • Hwabyung is a culture-bound anger syndrome in Korea. It manifests as one or more of a wide range of physical symptoms, in response to emotional stress. This case describes a 26-year old woman diagnosed as Hwabyung by HBDIS. She complained chiefly of binge eating and sleep disorder. She also complained of chest discomfort, abdominal discomfort, foreign body sensation in the throat, burning sensation and so on. We treated the patient with herbal medication, acupuncture, cupping, and moxa. After treatment, her condition improved. Standardized patients (SP) were used in teaching and practicing interviewing and physical examination skills for This report suggests that traditional Korean treatment such as herbal medication, acupuncture, cupping, moxa is effective for the treatment of Hwabyung patients.

A Convergence Study of Factors Influencing on Depressive Symptoms in Postmenopausal Middle-Aged Women (폐경 후 중년여성의 우울 증상 영향요인의 융합연구)

  • Kim, Younglan;Won, Mi Hwa
    • Journal of Convergence for Information Technology
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    • v.10 no.4
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    • pp.25-34
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    • 2020
  • The purpose of this study was to identify its influencing factors of depressive symptoms in postmenopausal middle-aged women though analyzing secondary data. The data of 647 with 40~65 aged women who participated in the Seventh Korea National Health and Nutrition Examination Survey in 2016 were used. The data were analyzed by complex sample independent t-test and multiple linear regression. In multiple linear regression analysis, economic status, subjective health status, perceived stress, smoking, physical activity, and limitation of active had significant effect on depressive symptoms in postmenopausal middle-aged women. These variables explained 36% of the variance in depressive symptoms. The findings suggest that healthcare providers should be considered to these factors when developing intervention strategies to decrease depressive symptoms in postmenopausal middle-aged women.

Effect of the Crude Polysaccharides Fraction from Eleutherococcus senticosus as a Immunoadjuvant to Soluble Antigens (BSA and OVA) (오가피 조다당의 단백질 항원 (BSA and OVA)에 대한 면역증강효과)

  • 황수현;하은숙;유광원;신광순;이상훈;이재권;이경호;윤택준;박우문
    • YAKHAK HOEJI
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    • v.47 no.3
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    • pp.167-175
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    • 2003
  • Eleutherococcus senticosus is a typical oriental folk medicinal herb. It has been used clinically as a anti-rheumatic disease, anti-stress, ischemic heart disease and gastric ulcer. In the present study, we examined the adjuvant activity of the crude polysaccharides fraction from Eleutherococcus senticosus, EN-3, on the induction of humoral and cellular immune responses against bovine serum albumin (BSA) or ovalbumin (OVA). The thioglycollate-induced macrophages and silica-induced dendritic-like cells cultured with BSA and EN-3 synergistically increased the production of TNF-$\alpha$ and IL-12. When mice were subcutaneously immunized with BSA + EN-3, the antibody titer against BSA was showed significantly higher than those immunized with BSA alone. In addition, when mice were immunized with OVA + FIA + EN-3, the antibody titer was showed similar patterns with the FCA. The assay for determining subisotype of antibody revealed that EN-3 augmented OVA-specific antibody titer of IgG1 and IgG2b. The culture supernatant obtained from splenocytes of mice treated with OVA + FIA + EN-3 also showed a higher level of both OVA-specific Th1-type (IL-2, IFN-${\gamma}$ and GM-CSF) and Th2-type cytokine (IL-4, IL-6 and IL-10). In vitro analysis of T cell proliferation to OVA on 8 weeks, the splenocytes of mice treated with OVA + EN-3 showed a significantly higher proliferating activity than those treated with OVA alone. These results suggest that EN-3 may possess adjuvant activities to potentially to enhance humoral as well as cellular immune response.

Olibanum-induced Apoptosis Signaling in Human Cervical Carcinoma HeLa Cells (자궁경부암세포(子宮經部癌細胞)(HeLa cell)에서 유향(乳香) 세포고사(細胞枯死) 기전(機轉) 연구(硏究))

  • Park, Kyung-Mi;Kong, Bok-Cheul;Lee, Su-Jung;Choe, Chang-Min;Yoo, Sim-Keun
    • The Journal of Korean Obstetrics and Gynecology
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    • v.19 no.2
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    • pp.92-106
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    • 2006
  • Purpose : To address the ability of Olibanum to induce cell death, we investigated the effect of olibanum on cell apoptosis. Twenty-four hours later, apoptosis occurred following olibanum exposure in a dose-dependent manner. Methods : We culture HeLa cell which is human metrocarcinoma cell in D-MEM included 10% fetal bovine serum(Hyclone Laboratories) below $37^{\circ}C$, 5% CO2. Then we observed apoptosis of log phage cell which is changed cultivation liquid 24 Hours periodically. Results : The treatment of BAPTA-AM regulated olibanum-induced apoptosis in HeLa human cervical carcinoma cells. The 24 hr-earlier -thapsigargin-pretreated cell showed the resistance against olibanum-induced apoptosis and the Ru360-mitochondrial uniporter-inhibited olibanum-induced apoptosis, too. It means that olibanum leads to the accumulation of calcium and the resultant apoptosis in HeLa cells. Immunoblotting data also shows that the expression of GRP78, ER stress marker protein, was induced by the olibanum. Bcl-2, anti-apototic protein, was decreased and that the expression of Bax, pro-apoptotic protein, was increased by the addition of olibanum. Interestingly, the olibanum increased the activity of caspase-8 as well as calpain cysteine pretense in HeLa cervical carcinoma cells. Calpain inhibitor-calpastatin as well as caspase-8C/A expression abrogated olibanum-induced apoptosis in the carcinoma cells. The inhibition of caspase-8 regulated olibanum-induced calpain activation but the inhibition of calpain did not have any effect on the caspase-8 activation in HeLa human cervical carcinoma cells. Conclusion : We conclude that olibanum induces the accumulation of calcium and the resultant apoptosis in which caspase-8 and calpain are involved.

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Effect of NADH-Dependent Enzymes Related to Oxygen Metabolism on Elimination of Oxygen-Stress of Bifidobacteria (NADH요구 산소대사관련 효소가 bifidobacteria의 산소스트레스 제거에 미치는 영향)

  • Ahn, Jun-Bae;Park, Jong-Hyun
    • Korean Journal of Food Science and Technology
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    • v.37 no.6
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    • pp.951-956
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    • 2005
  • Selection of oxygen-tolerant strains and elucidation of their oxygen tolerance mechanism were crucial for effective use of bifidobacteria. Oxygen-tolerant bifidobacteria were able to significantly remove environmental oxygen (oxygen removal activity) as compared to oxygen-sensitive strains. Most oxygen removal activity was inhibited by heat treatment and exposure to extreme pH (2.0) of bifidobacterial cell. NADH oxidase was major enzyme related to oxygen removal activity. Oxygen-tolerant bifidobacteria possessed high NADH peroxidase activity level to detoxify $H_2O_2$ formed from reaction of NADH oxidase. Addition of oxygen to anaerobic culture broth significantly increased activities of HADH oxidase and NADH peroxidase within 1hr and rapid increment of oxygen concentration was prevented. Results showed NADH oxidase and NADH peroxidase of oxygen-tolerant bifidobacteria played important roles in elimination of oxygen and oxygen metabolite $(H_2O_2)$.

Molecular Profiling of Rhizosphere Bacterial Communities Associated with Prosopis juliflora and Parthenium hysterophorus

  • Jothibasu, K.;Chinnadurai, C.;Sundaram, S.P.;Kumar, K.;Balachandar, D.
    • Journal of Microbiology and Biotechnology
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    • v.22 no.3
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    • pp.301-310
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    • 2012
  • Prosopis juliflora and Parthenium hysterophorus are the two arid, exotic weeds of India that are characterized by distinct, profuse growth even in nutritionally poor soils and environmentally stressed conditions. Owing to the exceptional growth nature of these two plants, they are believed to harbor some novel bacterial communities with wide adaptability in their rhizosphere. Hence, in the present study, the bacterial communities associated with the rhizosphere of Prosopis and Parthenium were characterized by clonal 16S rRNA gene sequence analysis. The culturable microbial counts in the rhizosphere of these two plants were higher than bulk soils, possibly influenced by the root exudates of these two plants. The phylogenetic analysis of V1_V2 domains of the 16S rRNA gene indicated a wider range of bacterial communities present in the rhizosphere of these two plants than in bulk soils and the predominant genera included Acidobacteria, Gammaproteobacteria, and Bacteriodetes in the rhizosphere of Prosopis, and Acidobacteria, Betaproteobacteria, and Nitrospirae in the Parthenium rhizosphere. The diversity of bacterial communities was more pronounced in the Parthenium rhizosphere than in the Prosopis rhizosphere. This culture-independent bacterial analysis offered extensive possibilities of unraveling novel microbes in the rhizospheres of Prosopis and Parthenium with genes for diverse functions, which could be exploited for nutrient transformation and stress tolerance in cultivated crops.

Flocculation of Red Tide Organisms in Sea Water by Using an Ignited Oyster Shell Powder and Loess Combination (소성굴패각분말과 황토의 동시 사용에 의한 적조생물의 응집)

  • KIM Sung-Jae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.36 no.6
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    • pp.716-722
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    • 2003
  • This study determined the optimum dosage for coagulation reactions of red tide organisms (RTO) using a combination of ignited oyster shell powder (10sp) and loess and examined the electrokinetic and rheological characteristics of their flocs. Two kinds of RTO, Cylindrotheca closterium and Skeletonema costatum, were sampled in Masan Bay and cultured in the laboratory. Coagulation experiments were conducted using various concentrations of IOSP, loess, IOSP+1oess, RTO, and a jar tester RTO cell numbers were counted for both the supernatant and RTO culture solution. The removal rates increased rapidly with increasing IOSP concentrations up to 50 mg/L and loess concentrations up to 800 mg/L. A removal rate of $100\%$ was reached at 400 mg/L of IOSP and 6,400 mg/L of loess. The highest increment $(16.7\%)$ of the rates of coagulation reaction occurred using both IOSP and loess (50+200 mg/L) in comparison with IOSP alone. The rate of coagulation reaction using both IOSP and loess (50+200 mg/L), $90.6\%,$ was similar to employing either IOSP of 150 mg/L or loess of 3,200 mg/L. All of the coagulation liquids for RTO, IOSP (200 mg/L), loess (200 ma/L), and IOSP+1oess (200+200 mg/L) revealed non-Newtonian fluid properties and therefore their shear rate vs. shear stress curves were non-linear. The coagulation liquids revealed elastic body properties at a lower shear rate increasing in the following order: RTO, IOSP (200 mg/L), loess (200 mg/L), and IOSP+1oess (200+200 mg/L. IOSP+1oess (200+200 mg/L) especially demonstrated plastic flow properties at a lower shear rate.

Cancer Chemopreventive Properties of Processed Ginseng

  • Surh, Young-Joon
    • Proceedings of the Ginseng society Conference
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    • 1998.06a
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    • pp.270-280
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    • 1998
  • Ginseng is one of the most widely used medicinal plants, particularly in East Asian countries. Certain fractions or purified ingredients of ginseng have been shown to exert inhibitory effects on growth of cancer cells in culture or on tumorigenesis in experimental animals. Moreover, a recent epidemiologic study reveals that ginseng intake is associated with a reduced risk for environmentally related cancers such as esophageal, gastric, colorectal, and pulmonary tumors. Heat treatment of Panax ginseng C. A. Meyer at the temperature higher than that applied to the conventional preparation of red ginseng yielded a mixture of saponins with potent antioxidative properties. Thus, the methanol extract of heat-processed ginseng (designated as'NGMe') attenuated lipid peroxidation in rat brain homogenates induced by ferric ion or ferric ion plus ascorbic acid. Furthermore, the extract protected against strand scission in f Xl 74 supercoiled DNA Induced by UV photolysis of H2O2 and was also capable of scavenging superoxide generated in vitro by xanthine/xanthine oxidate or in differentiated human promyelocytic leukemia (HL-60) cells by the tumor promoter,12-0-tetvade- canoylphorbol-13-acetate (TPA). Since tumor promotion is closely linked to oxidative stress, we have determined possible anti-tumor promotional effects of NGMe on two-stage mouse skin tumorigenesis. Topical application of NGMe onto shaven backs of female ICR mice 10 min prior to TPA significantly ameliorated skin papillomagenesi s initiated by 7,12-dimethylbenz (a) anthracene (DMBA).'Likewise, TPA-induced epidermal ornithine decarboxylase activity and elevation of tumor necrosis factor-a were suppressed signifies%fly by NGMe pretreatment. NGMe topically applied onto surface of hamster buccal pouch 10 min before each topical application of DMBA inhibited oral carcinogenesis by 76olo in terms of multiplicity. Taken together, these results suggest that processed Panax ginseng C. A. Meyer has potential cancer chemopreventive activities.

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