• Title/Summary/Keyword: degradation analysis

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NF-${\kappa}B$ Dependent IL-8 Secretion from Lung Epithelial Cells Induced by Peripheral Blood Monocytes Phagocytosing Mycobacterium Tuberculosis (결핵균을 탐석한 말초혈액단핵구 배양상층액에 의해 유도되는 폐상피세포주에서의 NF-${\kappa}B$ 의존성 IL-8 분비기전)

  • Park, Jae-Seuk;Jee, Young-Koo;Choi, Eun-Kyong;Kim, Keun-Youl;Lee, Kye-Young
    • Tuberculosis and Respiratory Diseases
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    • v.51 no.4
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    • pp.315-324
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    • 2001
  • Background : IL-8 is a potent chemotactic cytokine that plays an important role in the host defense mechanism against M. tuberculosis by recruiting inflammatory cells to the site of the infection. Lung epithelial cells, as well as alveolar macrophages are known to produce IL-8 in response to M. tuberculosis. IL-8 gene expression is mainly regulated on the level of transcription by NF-${\kappa}B$. This study investigated whether or not A549 cells produce IL-8 in NF-${\kappa}B$ dependent mechanism in response to macrophages phagocytosing M. tuberculosis. Methods : Peripheral blood monocytes that were obtained from healthy donors were cultured for 24 h with M. tuberculosis and a conditioned medium(CoMTB) was obtained. As a negative control, the conditioned medium without M. tuberculosis (CoMCont) was used. A549 cells were stimulated with M. tuberculosis, CoMCont and CoMTB and the IL-8 concentration in the culture media was measured by ELISA. The CoMTB induced IL-8 mRNA expression in the A549 cells was evaluated using RT-PCR, and CoMTB induced $I{\kappa}B{\alpha}$ degradation was measured using western blot analysis. CoMTB induced nuclear translocation and DNA binding of NF-${\kappa}B$ was also examined using an electrophoretic mobility shift assay(EMSA), and the CoMTB induced NF-${\kappa}B$ dependent IL-8 transcriptional activity was measured using a luciferase reporter gene assay. Results : CoMTB induced IL-8 production by A549 cells($46.8{\pm}4.8\;ng/ml$) was higher than with direct stimulation with M. tuberculosis ($6.8{\pm}2.9\;ng/ml$). CoMTB induced IL-8 mRNA expression increased after 2 h of stimulation and was sustained for 24 h. $I{\kappa}B{\alpha}$ was degraded after 10 min of CoMTB stimulation and reappeared by 60 min. CoMTB stimulated the nuclear translocation and DNA binding of NF-${\kappa}B$. The CoMTB induced NF-${\kappa}B$ dependent IL-8 transcriptional activity($13.6{\pm}4.3$ times control) was higher than either CoMCont($2.0{\pm}0.6$ times control) or M. tuberculosis ($1.4{\pm}0.6$ times control). Conclusion : A conditioned medium of peripheral blood monocytes phagocytosing M. tuberculosis stimulates NF-${\kappa}B$ dependent IL-8 production by the lung epithelial cells.

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The Effect of $I{\kappa}B{\alpha}$-SR Gene Transfer on the Sensitivity of Human Lung Cancer Cell Lines to Cisplatin and Paclitaxel ($I{\kappa}B{\alpha}$-SR 유전자이입이 Cisplatin, Paclitaxel에 대한 폐암세포주의 감수성에 미치는 영향)

  • Lee, Seok-Young;Seol, Ja-Young;Park, Kyung-Ho;Park, Gun-Min;Hwang, Yong-Il;Kim, Cheol-Hyeon;Jang, Seung-Hun;Kwon, Sung-Youn;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Lee, Choon-Taek
    • Tuberculosis and Respiratory Diseases
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    • v.51 no.2
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    • pp.122-134
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    • 2001
  • Background : Some chemotherapeutic drugs induce NF-${\kappa}B$ activation by degrading the $I{\kappa}B{\alpha}$ protein in cancer cells which contributes to anticancer drug resistance. We hypothesized that inhibiting $I{\kappa}B{\alpha}$ degradation would block NF-${\kappa}B$ activation and result in increased tumor cell mortality in response to chemotherapy. Methods : The "superrepressor" form of the NF-${\kappa}B$ inhibitor was transferred by an adenoviral vector (Ad-$I{\kappa}B{\alpha}$-SR) to the human lung cancer cell lines (NCI H157 and NCI H460). With a MIT assay, the level of sensitization to cisplatin and paclitaxel were measured. To confirm the mechanism, an EMSA and Annexin V assay were performed. Results : EMSA showed that $I{\kappa}B{\alpha}$-SR effectively blocked the NF-${\kappa}B$ activation induced by cisplatin. Transduction with Ad-$I{\kappa}B{\alpha}$-SR resulted in an increased sensitivity of the lung cancer cell lines to cisplatin and paclitaxel by a factor of 2~3 in terms of $IC_{50}$. Annexin-V analysis suggests that this increment in chemosensitivity to cisplatin probably occurs through the induction of apoptosis. Conclusion : The blockade of chemotherapeutics induced NF-${\kappa}B$ activation by inducing Ad-$I{\kappa}B{\alpha}$-SR, increased apoptosis and increasing the chemosensitivity of the lung cancer cell lines tested, subsequently. Gene transfer of $I{\kappa}B{\alpha}$-SR appears to be a new therapeutic strategy of chemosensitization in lung cancer.

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The Role of Heme Oxygenase-1 in Lung Cancer Cells (폐암세포주에서 Heme Oxygenase-1의 역할)

  • Jung, Jong-Hoon;Kim, Hak-Ryul;Kim, Eun-Jung;Hwang, Ki-Eun;Kim, So-Young;Park, Jung-Hyun;Kim, Hwi-Jung;Yang, Sei-Hoon;Jeong, Eun-Taek
    • Tuberculosis and Respiratory Diseases
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    • v.60 no.3
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    • pp.304-313
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    • 2006
  • Background : Heme oxygenase-1 (HO-1) is an inducible enzyme that catalyzes the oxidative degradation of heme to form biliverdin, carbon monoxide (CO), and free iron. The current evidence has indicated a critical role of HO-1 in cytoprotection and also in other, more diverse biological functions. It is known that the high expression of HO-1 occurs in various tumors, and that HO-1 has an important role in rapid tumor growth because of its antioxidative and antiapoptotic effects. Therefore, the role of HO-1 was analyzed in human lung cancer cell lines, and especially in the A549 cell line. Material and Methods : Human lung cancer cell lines, i.e., A549, NCI-H23, NCI-H157 and NCI-H460, were used for this study. The expression of HO-1 in the untreated state was defined by Western blotting. ZnPP, which is the specific HO inhibitor we used, and the viability of cells were tested for by conducting MTT assaysy. The HO enzymatic activity, as determined via the bilirubin level, was also indirectly measured. Moreover, the generation of intracellular hydrogen peroxide (H2O2) was monitored fluorimetrically with using a scopoletin-horse radish peroxidase (HRP) assay and 2',7'-dichlorofluorescein diacetate (DCFH-DA). We have also transfected small HO-1 interfering RNA (siRNA) into A549 cells, and the apoptotic effects were evaluated by flow cytometric analysis and Western blotting. Results : The A549 cells had a greater expression of HO-1 than the other cell lines, whereas ZnPP significantly decreased the viability of the A549 cells more than the viability of the other lung cancer cells in a dose-dependant fashion. Consistent with the viability, the HO enzymatic activity also was decreased. Moreover, intracellular H2O2 generation via ZnPP was induced in a dose-dependent manner. Apoptotic events were, then induced in the HO-1 siRNA transfected A549 cells. Conclusion : HO-1 provides new important insights into the possible molecular mechanism of the antitumor therapy in lung cancer.

A Study for Perception of Hair Damage Using Friction Coefficient of Human Hair (모발의 마찰계수를 통한 모발 손상 인식 연구)

  • Lim, Byung Tack;Seo, Hong An;Song, Sang-Hun;Son, Seong Kil;Kang, Nae-Gyu
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.46 no.3
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    • pp.295-305
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    • 2020
  • Treatment for beauty using oxidizing agents damages hair with inducing structural alteration in cuticle layer, degradation of protein, and loss of lipid. This study connects a frictional coefficient upon the damaged hair by an instrumental test to the texture test by human being, and considered a moisture as a factor of the damage. A friction coefficient has been measured upon the hair with successive treatment of dye, perm, and bleach. The friction coefficient from the hair dye-treated three times was defined with 0.60, where 58% of answerer indicated an initial damage point as the hairs of iteration of dye-treatment increased. Even bleach treated three times results in 0.84 of friction coefficient corresponding to 88% of answerer attributed the hair to an initially damaged hair. In order to figure out a lipid loss in hair for human being to respond damage, a friction coefficient of the hair was controlled by removing 18-methyleicosanoic acid (18-MEA). The initial damage has been recognized by 0.60 of the friction coefficient for the 68% of answerer. Since moisture is the largest portion of the components in hair, moisture analysis has been performed to study a relationship between texture of damage and the friction coefficient from an instrumental evaluation. As an iteration of dye increases, the hair became hydrophilic with smaller contact angle. It is found that a damaged hair by dyeing possessed more than 0.42% of moisture compared to a healthy hair. Finally, it is elucidated that an increase of moisture in hair induced higher adhesive force corresponding to the friction coefficient, and the friction coefficient above 0.6 is attributed to the preception of hair damage.

Apoptotic Effect of Co-Treatment with Valproic Acid and 17AAG on Human Osteosarcoma Cells (Valproic acid와 17AAG의 병용처리가 사람골육종세포에 미치는 세포자멸사 효과에 대한 연구)

  • Park, Jun-Young;Park, Se-Jin;Kim, In-Ryoung;Park, Bong-Soo;Jeong, Sung-Hee;Ko, Myung-Yun;Ahn, Yong-Woo
    • Journal of Oral Medicine and Pain
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    • v.36 no.1
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    • pp.11-20
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    • 2011
  • Valproic acid (VPA) is a well-known anticonvulsive agent and has been used in the treatment of epilepsy for almost 30 years. VPA emerged in 1997 as an antineoplastic agent. And it is known that antitmor activity of VPA is associated with its targeted at histone deacetylases. 17AAG, Inhibition of HSP90 leads to the proteasome degradation of the HSP90 client proteins, such as Akt, Raf/Ras, Erk, VEGF, cyclin D and p53, and causes potent antitumor activity. It is reported that 17AAG-induced HSP90 inhibition results in prevention of cell proliferation and induction of apoptosis in several types of cancer. This study was undertaken to investigate the synergistic apoptotic effect of co-treatment with the histone deacetylases inhibitor, VPA and the HSP90 inhibitor, 17AAG on human osteosarcoma (HOS) cells. Cell viability was evaluated by trypan-blue exclusion. Induction and augmentation of apoptosis were confirmed by Hoechst staining, flow cytometry (DNA hypoploidy and MMP change), Westen blot analysis and immunofluorescent staining. In this study, HOS cells co-treated with VPA and 17AAG showed several lines of apoptotic manifestation such as nuclear condensations, the reduction of MMP, the decrease of DNA content, the release of cytochrome c into cytosol, the translocation of AIF onto nuclei, and activation of caspase-3, caspase-7 and PARP whereas each single treated HOS cells did not. Although the single treatment of 1 mM VPA or 0.5 ${\mu}M$ 17AAG for 48 h did not induce apoptosis, the co-treatment with them induced prominently apoptosis. Therefore our data in this study provide the possibility that combination therapy with VPA and 17AAG could be considered as a novel therapeutic strategy for human osteosarcoma.

Effect of Temperature on Amino-Carbonyl Reaction (Amino-Carbonyl 반응(反應)에 미치는 온도(溫度)의 영향(影響))

  • Kim, Yong-Nyun;Kim, Chang-Mok;Han, Kang-Wan;Oh, Sung-Ki
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.11 no.1
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    • pp.51-56
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    • 1982
  • The thermal degradation of 0.05M glucose-arginine model system was occurred during heat treatment for 0$\sim$7 hours at $60{\sim}120^{\circ}C$. and the melanoid in formation was investigated as a function of temperature. The decomposition reaction of glucose and arginine, as well as the reaction of melanoidin formation, followed first-order kinetics, except the reaction at $120^{\circ}C$. and the rate constants ($hr^{-1}\times 10^3$) of those reactions were ranged from 14.20 to 837. 10. Temperature dependence of the rate constants was characterized by the Arrhenius equation, except the reaction at $120^{\circ}C$. The ranges of activation energy and $Q_{10}$ values were 12.122$\sim$18.142 kcal/mole and 1.65$\sim$2.12, respectively.

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The Antimicrobial Characteristics of McSSP-31 Purified from the Hemocyte of the Hard-shelled Mussel, Mytilus coruscus (참담치(Mytilus coruscus) 혈구(hemocyte)에서 분리한 McSSP-31의 항균 특성 분석)

  • Oh, Ryunkyoung;Lee, Min Jeong;Kim, Young-Ok;Nam, Bo-Hye;Kong, Hee Jeong;Kim, Joo-Won;Park, Jung-Youn;Seo, Jung-Kil;Kim, Dong-Gyun
    • Journal of Life Science
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    • v.27 no.11
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    • pp.1276-1289
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    • 2017
  • This study isolated and purified the antimicrobial peptide McSSP-31 from an acidified hemocyte extract of a Mytilus coruscus. The antimicrobial peptide was purified by using a $C_{18}$ reversed-phase high-performance liquid chromatography (HPLC). The peptide was determined to be 3330.549 Da by matrix assisted-laser desorption ionization time-of-flight mass spectrophotometer (MALDI-TOF/MS). The N-terminus of a 14 amino-acid sequence was identified as P-S-P-T-R-R-S-T-S-R-S-K-S-R by Edman degradation method. The acquired sequence showed a 93% similarity with the sperm-specific protein Phi-1, which is from M. californianus. The identified open-reading frame (ORF) of peptide was 306 bp encoding 101 amino acids, which was analyzed by rapid amplification of cDNA ends (RACE), cloning and sequencing analysis. We compared the full sequence with other known proteins that reveal the sperm-specific protein Phi-1 (93.5%) of M. californianus. Synthesized antimicrobial peptide (McSSP-31) showed antibacterial activity against gram-positive bacteria including B. subtilis, S. mutans, S. aureus and gram-negative bacteria including E. coli, K. pneumoniae, P. mirabilis, P. aeruginosa and fungi, C. albicans. Also, synthesized peptide showed strong antibacterial activity against antibiotic-resistant strains, including S. aureus. The cytotoxicity of the peptide was determined by using the HUVEC human cell line. The peptide did not exhibit any significant cytotoxic effects on the normal human cell line, and it had very low hemolytic activity with flounder hemoglobin. The results demonstrated that peptide purified from the hemocyte of a M. coruscus exhibits antibacterial activity against various bacteria and has the potential to be an alternative antibiotic agent.

Diversity, Saccharification Capacity, and Toxigenicity Analyses of Fungal Isolates in Nuruk (누룩곰팡이 분리균의 다양성 및 당화능 분석과 독소생산능 조사)

  • Kim, Min Sik;Kim, Sinil;Ha, Byeong-Seok;Park, Hye-Young;BaeK, Seong-Yeol;Yeo, Soo-Hwan;Ro, Hyeon-Su
    • The Korean Journal of Mycology
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    • v.42 no.3
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    • pp.191-200
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    • 2014
  • Nuruk samples collected from various regions in Korea were investigated in terms of fungal contents and diversity. In measurement of colony forming unit (CFU) in Nuruk suspensions on DRBC agar, Nuruk samples MS4, MS8, and MS10 were among the highest fungal density, with $1,278.9{\pm}21.6$ (${\times}10^4$), $1,868.0{\pm}27.7$ (${\times}10^4$), and $775.1{\pm}19.2$ (${\times}10^4$) were among the samples showing the highest fungal density. CFU per 20 mg Nuruk, respectively. The majority of fungal components were yeasts, including Pichia anomala, P. kudriavzevii, Kluyveromyces marxianus, and Saccharomycopsis fibuligera, whereas Aspergillus oryzae and Rhizopus oryzae, the representative Nuruk fungi, were predominant only in the low fungal density Nuruks (MS2, MS5, and MS11). Saccharification capability of the fungal isolates was assessed by measurement of amylase activity in the culture broth. The highest amylase activity was found in A. niger and A. luchuensis, followed by S. fibuligera. A. oryzae and R. oryzae showed fair amylase activity but significantly lower than those of the three fungal species. R. oryzae was suggested to play an additional role in degradation of ${\beta}$-glucan in crop component of Nuruk since R. oryzae was the only fungus that showed ${\beta}$-glucanase activity among the fungal isolates. To confirm the safety of Nuruk, aflatoxigenicity of the isolated Aspergillus was estimated using the DNA markers norB-cypA, aflR, and omtA. All of the isolates turned out to be non-aflatoxigenic as evidenced by the deletion of gene markers, norB-cypA and aflR, and the absence of aflatoxin in the culture supernatants shown by TLC analysis.

Physiological and Biochemical Characterization of Bacillus spp. from Polychaete, Perinereis aibuhitensis (갯지렁이(Perinereis aibuhitensis)에서 분리한 Bacillus spp.의 생리생화학적 특성 분석)

  • Shin, Seyeon;Yundendorj, Khorloo;Lee, Sang-Suk;Kang, Kyoung-Ho;Kahng, Hyung-Yeel
    • Journal of Life Science
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    • v.23 no.3
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    • pp.415-425
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    • 2013
  • This study compared the characteristics of five Bacillus strains capable of aerobic and anaerobic growth, CBW3, CBW4, CBW9, CBW14 and EBW10. They were isolated and selected from a polychaete, Perinereis aibuhitensis, which is known as a good degrader of organic compounds in marine wetland. Based on a 16S rRNA sequence, CBW3 and CBW14 were found to share more than 99.8% similarity with B. nanhaiensis, B. arsenicus and B. barbaricus. CBW4, CBW9 and EBW10 shared 92.7%, 99.8%, and 99.8% similarity with B. anthracis, B. algicoa and B. thuringiensis, respectively. The temperature, salinity, and pH ranges of the cell growth of the Bacillus strains were $4-45^{\circ}C$, 0-17%, and pH 5-pH 9, respectively. All Bacillus strains were found to exhibit enzyme activities for the degradation of casein and starch. Notably, strain EBW10 exhibited the enzyme activities for all the tested macromolecules, DNA, casein, starch, cellulose, and four kinds of Tweens, which suggests the possibility that it had protease, amylase, cellulose, and lipase. All five Bacillus strains had alkaline phosphatase activities, and the strains CBW3, CBW4, and EBW10 also had acid phospatase. Strains CBW3 and EBW10 exhibited the enzyme activities both for esterase (C4) and esterase lipase (C8). The analysis of fatty acids revealed that in all strains, major fatty acids were anteiso $C_{15:0}$ and iso $C_{15:0}$.

Cartoon Criticism; The subject and the gaze based on Lacan' s theory otherness of vision : focusing on KUBRICK of Kang, Do-Ha (라캉의 시각의 타자성(대상 a)에 근거한 만화비평으로서의 주체와 응시 : 강도하의 큐브릭을 중심으로)

  • Yang, Seung-Kyu
    • Cartoon and Animation Studies
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    • s.26
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    • pp.79-108
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    • 2012
  • This thesis is on the purpose of seeking for the possibility of a mechanism in interpreting, analyzing and criticizing cartoons which are applied to the concept of "the gaze" based on "the otherness of vision", which states a pluralistic visual world. It proves that cartoons are in line with other art works that are the subject to "lack and desire"; the gaze, greets reality and acts as an important criterion of analyzing and criticizing the trend in contemporary art, in which the cartoon expresses the gaze in harmony within its work of art. In this thesis, the artist, Kang Doh-ha structuralized ambiguous and difficult forms of art as he has cumulated experimental minds by working in an indie cartoon plane for a long period of time. Among his works of art, he identified the "invisible world" through his piece "Kubrik". Therefore, he represented: a metaphor and a metonymy, an ambiguously situational expression, an intentional and emotional error, the structure of individuality and integration, and finally tension beyond its meaning through use of 'the gaze' that is both the cause and the subject of a desire in the visible world which Lacan academized when he interpreted and analyzed "Kubrik". The concept of the gaze can be used in a variety of ways to display one another's presence in relation each character, revealing a spot of lack by staring back at readers or audiences and furthermore, to analyze and criticize the hidden side of the art piece by critics. The most important details are the artist's gaze, which is seen in the eyes of the analysis and also his or her criticism of the cartoon, which functions as a metaphoric screen in which the subject himself or herself betrays the law of desires thus enabling the violent and cruel reality to be masked and indulged in plays. This will serve as an element that will lead into an art as well as control the degradation to just a piece of enjoyment with the cartoon remaining only within the visual world.