• 제목/요약/키워드: Cell manipulation

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Enhanced Lycopene Production by UV-C Irradiation in Radiation-Resistant Deinococcus radiodurans R1

  • Kang, Chang Keun;Yang, Jung Eun;Park, Hae Woong;Choi, Yong Jun
    • Journal of Microbiology and Biotechnology
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    • v.30 no.12
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    • pp.1937-1943
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    • 2020
  • Although classical metabolic engineering strategies have succeeded in developing microbial strains capable of producing desired bioproducts, metabolic imbalance resulting from extensive genetic manipulation often leads to decreased productivity. Thus, abiotic strategies for improving microbial production performance can be an alternative to overcome drawbacks arising from intensive metabolic engineering. Herein, we report a promising abiotic method for enhancing lycopene production by UV-C irradiation using a radiation-resistant ΔcrtLm/crtB+dxs+ Deinococcus radiodurans R1 strain. First, the onset of UV irradiation was determined through analysis of the expression of 11 genes mainly involved in the carotenoid biosynthetic pathway in the ΔcrtLm/crtB+dxs+ D. radiodurans R1 strain. Second, the effects of different UV wavelengths (UV-A, UV-B, and UV-C) on lycopene production were investigated. UV-C irradiation induced the highest production, resulting in a 69.9% increase in lycopene content [64.2 ± 3.2 mg/g dry cell weight (DCW)]. Extended UV-C irradiation further enhanced lycopene content up to 73.9 ± 2.3 mg/g DCW, a 95.5% increase compared to production without UV-C irradiation (37.8 ± 0.7 mg/g DCW).

HSP27 EXPRESSION IN OSTEOBLAST BY THERMAL STRESS (골모세포에서 열자극에 의한 Hsp27 발현에 대한 연구)

  • Rim, Jae-Suk;Kim, Byeong-Ryol;Kwon, Jong-Jin;Jang, Hyon-Seok;Lee, Eui-Suk;Jun, Sang-Ho;Woo, Hyeon-Il
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.30 no.1
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    • pp.11-21
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    • 2008
  • Aim of the study: Thermal stress is a central determinant of osseous surgical outcomes. Interestingly, the temperatures measured during endosseous surgeries coincide with the temperatures that elicit the heat shock response of mammalian cells. The heat shock response is a coordinated biochemical response that helps to protect cells from stresses of various forms. Several protective proteins, termed heat shock proteins (hsp) are produced as part of this response. To begin to understand the role of the stress response of osteoblasts during surgical manipulation of bone, the heat shock protein response was evaluated in osteoblastic cells. Materials & methods: With primary cell culture studies and ROS 17/2.8 osteoblastic cells transfected with hsp27 encoding vectors culture studies, the thermal stress response of mammalian osteoblastic cells was evaluated by immunohistochemistry and western blot analysis. Results: Immunocytochemistry indicated that hsp27 was present in unstressed osteoblastic cells, but not fibroblastic cells. Primarily cultured osteoblasts and fibroblasts expressed the major hsp in response to thermal stress, however, the small Mr hsp, hsp27 was shown to be a constitutive product only in osteoblasts. Creation of stable transformed osteoblastic cells expressing abundant hsp27 protein was used to demonstrate that hsp27 confers stress resistance to osteoblastic cells. Conclusions: The demonstrable presence and function of hsp27 in cultured bones and cells implicates this protein as a determinant of osteoblastic cell fate in vivo.

Rapid Measurement of $NH_3$ and Weak Acid Permeation Through Liposomes and Renal Proximal Tubule Membranes

  • Bae, Hae-Rahn;Suh, Duck-Joon;Lee, Sang-Ho
    • The Korean Journal of Physiology
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    • v.28 no.2
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    • pp.203-214
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    • 1994
  • Using the methods of stopped-flow and epifluorescence microscopy with entrapped fluorophore, membrane permeability of $NH_3$ and weak acids in liposomes, renal brush border (BBMV) and basolateral membrane vesicles (BLMV), and primary culture cells from renal proximal tubule was measured. Permeability coefficient (cm/sec) of $NH_3$ was $(2.9{\times}10^{-2}$ in phosphatidylcholine liposome $25^{\circ}C)$, $5.9{\times}10^{-2}$ in renal proximal tubule cell $(37^{\circ}C)$, $4.0{\times}10^{-2}\;and\;2.4{\times}10^{-2}$ in BBMV and BLMV $(25^{\circ}C)$, respectively. Formic acid has the highest permeability coefficient among the weak acids tested, which was $4.9{\times}10^{-3}$ in liposome, $5.0{\times}10^{-3}$ in renal proximal tubule cell, $9.1{\times}10^{-3}$ in BBMV and $3.8{\times}10^{-3}$ in BLMV. There was a linear relationship between external concentration of nonionized formic acid and initial rate of flux of formic acid in liposome, and the slope coincided with the value of permeability coefficient of formic acid measured in pH 7.0. These results show that techniques of stopped-flow and epifluorescence microscopy with entrapped fluorophore provide the precise method of measurement of very rapid transport of nonelectrolytes through membranes with the advantages of instantaneous mixing effect, good resolution time and easy manipulation.

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Histological Changes of the Acupoint by Acupuncture Stimulation (침자(鍼刺)한 경혈(經穴) 부위(部位)의 조직학적 변화(組織學的 變化))

  • Kim, Jeong-Sang;Park, Min-Hee;Cho, Kwang-Phil;Jung, Hae-Man
    • Applied Microscopy
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    • v.30 no.1
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    • pp.81-87
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    • 2000
  • This study has been carried out to understand histochemical and ultrastructural changes by acupuncture stimulation at the acupoint of the Zusanli (St. 36) in the rat. A lot of mast cells are observed in the peripheral of the hole with a sparrow-pecking and twistin -needle manipulation to the acupoint Zusanli for induction of 'Qi'. These cells contained a lot of granules with varied electron density and exocytosis granules were observed near the mast cell. H-bands of the muscle fiber that situated near the hole were shortened. It is assumed that these muscles are contracted by acupuncture stimulation. These results imply that functional relationship between mast cells in the dermis and Qi-sensation induced by acupuncture plays an important role on the specific receptor response to the mechanical stimulation.

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Development of Host-Vector Systems for Lactic Acid Bacteria (유산균의 Host-Vector System 개발)

  • 윤성식;김창민
    • Microbiology and Biotechnology Letters
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    • v.29 no.1
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    • pp.1-11
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    • 2001
  • Lactic acid bacteria (LAB) are widely used for various food fermentation. With the recent advances in modern biotechnology, a variety of bio-products with the high economic values have been produced using microorganisms. For molecular cloning and expression studies on the gene of interest, E. coli has been widely used mainly because vector systems are fully developed. Most plasmid vectors currently used for E, coli carry antibiotic-resistant markers. As it is generally believed that the antibiotic resistance markers are potentially transferred to other bacteria, application of the plasmid vectors carrying antibiotic resistance genes as selection markers should be avoided, especially for human consump-tion. By contrast, as LAB have some desirable traits such that the they are GRAS(generally recognized as safe), able to secrete gene products out of cell, and their low protease activities, they are regarded as an ideal organism for the genetic manipulation, including cloning and expression of homologous and heterologous genes. However, the vec-tor systems established for LAB are stil insufficient to over-produce gene products, stably, limiting the use of these organisms for industrial applications. For a past decade, the two popular plasmid vectors, pAM$\beta$1 of Streptococcus faecalis and pGK12 theB. subtilis-E. coli shuttle vector derived from pWV01 of Lactococcus lactis ssp. cremoris wg 2, were most widely used to construct efficient chimeric vectors to be stably maintained in many industrial strains of LAB. Currently, non-antibiotic markers such as nisin resistance($Nis^{r}$ ) are explored for selecting recombi-nant clone. In addition, a gene encoding S-layer protein, slp/A, on bacterial cell wall was successfully recombined with the proper LAB vectors LAB vectors for excretion of the heterologous gene product from LAB Many food-grade host vec-tor systems were successfully developed, which allowed stable integration of multiple plasmid copies in the vec-mosome of LAB. More recently, an integration vector system based on the site-specific integration apparatus of temperate lactococcal bacteriophage, containing the integrase gene(int) and phage attachment site(attP), was pub-lished. In conclusion, when various vector system, which are maintain stably and expressed strongly in LAB, are developed, lost of such food products as enzymes, pharmaceuticals, bioactive food ingredients for human consump-tion would be produced at a full scale in LAB.

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Robot Hopscotch Game Using Mobile Robots (이동 로봇을 사용한 로봇 팔방 놀이)

  • Ko, Nak-Yong;Moon, Yong-Seon;Bae, Young-Chul
    • The Journal of the Korea institute of electronic communication sciences
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    • v.6 no.2
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    • pp.296-303
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    • 2011
  • This paper describes a game called the "robot hopscotch game" which uses mobile robots. In this game, a mobile robot moves on a board which consists of a number of square cells. This is similar to the usual hopscotch game in which a human player hops through a course on the ground. In the robot hopscotch game, each cell has a visual light communication unit. The unit sends identification signal and the robot which moves over the cell receives the signal for location recognition. In the game, a human player or a human operator competes with a robot or other human player on how accurately and fast they move to a goal location passing through the given way point cells. People played the game at the RoboWorld 2008 which was held at COEX Seoul, on 16th to 20th of October 2008. The replies to the questionnaire to the users show that users have interests on the game and expect further expansion of robot use though they are not in favor of the robot's outlook and they feel inconvenience on manipulation of the robot.

Development of a real-time Analysis System of Microchip Fluorescence Images based on Server-Client (서버 클라이언트 기반의 실시간 마이크로칩 형광 이미지 분석 시스템 개발)

  • Cho, Migyung;Shim, Jaesool
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.17 no.5
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    • pp.1239-1244
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    • 2013
  • In the field of clinical medicine and research, the analysis of such as protein and DNA at the molecular level and even at the cell level are necessary for disease diagnosis and treatment. In many cases, a real time image of samples is needed for the accurate analysis and manipulation of samples since experimental samples are degenerated with time. In this research, a three-dimensional fluorescence microscope device was developed for taking images of protein and DNA inside a single cell and the server-client based image analysis system was made for an integrated management of the real-time images taken from the microscope device. The system consists of a fluorescent measurement device, the associated software and a client program on smartphone. The developed system allows doctors or experimental managers to receive and look at the real-time experimental images taken from the samples of patients anywhere in the emergency, to analyze results and to instantly diagnose the disease and to transfer the results to the patients. As a result, the system is able to be utilized in the implementation of ubiquitous health as well.

Hepatoprotective Effect of Manual Acupuncture at Sam-hwang Points on $CCl_4$-induced Liver Damage in Rats ($CCl_4$로 유발된 흰쥐의 간손상에 대한 삼황혈(三黃穴) 수기침자극(手技鍼刺戟)의 간기능 보호효과)

  • Lee, Hyun;Hong, Kwon-Eui;Kim, Young-Il;Kim, Tae-Han;Choi, Hak-Joo;Kim, Sun-Jin;Yim, Yun-Kyoung
    • Korean Journal of Acupuncture
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    • v.22 no.1
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    • pp.109-116
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    • 2005
  • Objectives : To investigate the hepatoprotective effect of manual acupuncture at Sam-hwang points on hepatotoxicity in $CCl_4$-intoxicated rats. Methods : Rats were injected intraperitoneally with $CCl_4$(1ml/kg) and treated with manual acupuncture at left Sam-hwang points 3 times a week for 10 weeks. A non-acupoint in left Gluteal area was selected as a sham point. Acupuncture was performed by an experienced acupuncture doctor using tonifying manipulation technique. To estimate the effects on hepatotoxici쇼, body weight and liver weight of rats were measured, and biochemical assays(ALT, AST, ALP, Total cholesterol, etc.) and blood cell analysis were performed. Results : Manual acupuncture with tonifying technique at Sam-hwang points reduced liver weight, ALT, AST, ALP activities and total cholesterol in serum significantly compared with control group. In blood cell analysis, WBC and lymphocytes were reduced but neutrophil was increased by manual acupuncture at Sam-hwang points in rats. Conclusions : Manual acupuncture with tonifying technique at Sam-hwang points has an hepatoprotective effect on $CCl_4$-induced liver damage in rats.

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Antifungal Mechanism of Action of Lauryl Betaine Against Skin-Associated Fungus Malassezia restricta

  • Do, Eunsoo;Lee, Hyun Gee;Park, Minji;Cho, Yong-Joon;Kim, Dong Hyeun;Park, Se-Ho;Eun, Daekyung;Park, Taehun;An, Susun;Jung, Won Hee
    • Mycobiology
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    • v.47 no.2
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    • pp.242-249
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    • 2019
  • Betaine derivatives are considered major ingredients of shampoos and are commonly used as antistatic and viscosity-increasing agents. Several studies have also suggested that betaine derivatives can be used as antimicrobial agents. However, the antifungal activity and mechanism of action of betaine derivatives have not yet been fully understood. In this study, we investigated the antifungal activity of six betaine derivatives against Malassezia restricta, which is the most frequently isolated fungus from the human skin and is implicated in the development of dandruff. We found that, among the six betaine derivatives, lauryl betaine showed the most potent antifungal activity. The mechanism of action of lauryl betaine was studied mainly using another phylogenetically close model fungal organism, Cryptococcus neoformans, because of a lack of available genetic manipulation and functional genomics tools for M. restricta. Our genome-wide reverse genetic screening method using the C. neoformans gene deletion mutant library showed that the mutants with mutations in genes for cell membrane synthesis and integrity, particularly ergosterol synthesis, are highly sensitive to lauryl betaine. Furthermore, transcriptome changes in both C. neoformans and M. restricta cells grown in the presence of lauryl betaine were analyzed and the results indicated that the compound mainly affected cell membrane synthesis, particularly ergosterol synthesis. Overall, our data demonstrated that lauryl betaine influences ergosterol synthesis in C. neoformans and that the compound exerts a similar mechanism of action on M. restricta.

A Temperature-Dependent Index of Mitotic Interval ($\tau_0$) in Haliotis gigantea and Haliotis discus

  • Park, In-Seok;Im, Jae-Hyun;Lee, Young-Don;Kim, Bong-Lae;Han, Seock-Jung
    • Proceedings of the Korean Aquaculture Society Conference
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    • 2003.10a
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    • pp.29-29
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    • 2003
  • In order to establish effective procedures for chromosome manipulation in Haliotis gigantea and H. discus, which are of enormous aquacultural potential, temperature-dependent measures of mitotic intervals ($\tau$$_{0}$) were determined. Mitotic intervals ($\tau$$_{0}$) in these abalone were determined by averaging the duration of the first and third embryonic divisions over a range of temperatures from 8 to 26$^{\circ}C$. The relationships of each mitotic interval at two cell ($\tau$$_{I}$), four cell ($\tau$$_{II}$ ), eight cell ($\tau$$_{III}$), sixteen cell ($\tau$$_{IV}$ ) and $\tau$$_{0}$, to temperature (T in $^{\circ}C$) in H. gigantea were log $\tau$$_{I}$ : 176.1-28.3T, log $\tau$$_{II}$ : 199.5-12.4T, log $\tau$$_{III}$ = 236.2-12.2T, log $\tau$$_{IV}$ = 269.3-14.lT and log $\tau$$_{0}$ : 83.1-32.8, respectively. The relationships of each mitotic interval at $\tau$$_{I}$, $\tau$$_{II}$ , $\tau$$_{III}$, $\tau$$_{IV}$ and $\tau$$_{0}$, to temperature in H. discus were log $\tau$$_{I}$ = 104.9-13.8T, log $\tau$$_{II}$ : 138.3-10.5T, $\tau$$_{III}$ : 172.4-10.2T, log $\tau$$_{IV}$ : 211.3-12.2T and log $\tau$$_{0}$=85.6-33.3T, respectively. There were strong, negative correlations between mitotic interval and water temperatures for all ten temperatures in these two species (H. gigantea: Y = -138.75 logX + 341.25, $R^2$ = 0.97; H. discus: Y = -112.33 logX + 255.22, $R^2$ = 0.98, where Y is mitotic interval and X is temperature).d X is temperature).rature).

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