• Title/Summary/Keyword: Manipulation

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Production of Gamma-Linolenic Acid in Pichia pastoris by Expression of a Delta-6 Desaturase Gene from Cunninghamella echinulata

  • Wan, Xia;Zhang, Yinbo;Wang, Ping;Huang, Fenghong;Chen, Hong;Jiang, Mulan
    • Journal of Microbiology and Biotechnology
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    • v.19 no.10
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    • pp.1098-1102
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    • 2009
  • Gamma-linolenic acid (GLA, C18:3 ${\Delta}^{6,9,12}$) is synthesized by a delta-6 fatty acid desaturase using linoleic acid (LA, C18:2 ${\Delta}^{9,12}$) as a substrate. To enable the production of GLA in the conventional yeast Pichia pastoris, we have isolated a cDNA encoding the delta-6 fatty acid desaturase from Cunninghamella echinulata MIAN6 and confirmed its function by heterogeneous expression in P. pastoris. Sequence analysis indicated that this cDNA sequence has an open reading frame of 1,404 bp, which encodes a 52 kDa peptide of 468 amino acids. This sequence has 64% identity to the previously reported delta-6 fatty acid desaturase from Rhizopus oryzae. The polypeptide has a cytochrome b5 domain at the N-terminus including the HPGG motif in the heme-binding region, as reported for other delta-6 fatty acid desaturases. In addition, this enzyme differs from other desaturases by the presence of three possible N-linked glycosylation sites. Analysis of the fatty acid composition demonstrated the accumulation of GLA to the level of 3.1% of the total fatty acids. Notably, the amounts of ginkgolic acid (C17:1) and palmitic acid (C16:0) were increased from 1.3% to 29.6% and from 15% to 33%, respectively. These results reveal that the modification of the fatty acid biosynthetic pathway by genetic manipulation in order to produce specific polyunsaturated fatty acids in P. pastoris is a promising technique.

Mesenteric Lymphatic Delivery of Oral Anticancer Tegafur by Emulsion Formulations (유제화에 의한 경구용 항암제인 테가푸르의 장관 임파수송)

  • Lee, Yong-Bok;Nam, Kweon-Ho;Chang, Woo-Ik;Oh, In-Joon;Koh, Ik-Bae
    • Journal of Pharmaceutical Investigation
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    • v.25 no.1
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    • pp.55-62
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    • 1995
  • W/O and O/W emulsions of tegafur (50 mg/5 ml/kg) were orally administered to rats to compare with their mesenteric lymphatic delivery effects. And also in order to demonstrate the lymph targeting associated to the oral route, it was deemed necessary to investigate the fate of solution after oral administration as a control. Lymph and plasma samples were periodically taken from each subject of mesenteric lymphatic duct cannulated rats. Then, lymph and plasma levels of tegafur and its active metabolite, 5-FU, were simultaneously observed. Also pharmacokinetic parameters were compared with each others. On the other hand, most previous studies of lymphatic transport have not addressed the question of whether an increase in mesenteric or thoracic lymph transport by the manipulation of a suspected variable was due to a selective delivery to the intestinal lymphatics or an overall increase availability. Therefore, based on a physiologically based pharmacokinetic model which represents the characteristics of lymphatic systems, we are also going to determine the contributions of mesenteric lymph transport versus thoracic lymph transport of tegafur reported in reference(13). In comparison with tegafur solution, AUC and mean residence time of plasma tegafur were significantly increased in W/O emulsion but significantly decreased in O/W emulsion. Lymph flow rates were similar in both solution and W/O emulsion but half in O/W emulsion. AUC of tegafur in mesenteric lymph and in plasma for W/O emulsion were 3.7 times and 2.9 times more than those for O/W emulsion, respectively. And AUC of 5-FU in thoracic lymph for W/O emulsion was 3.7 times more than that for O/W emulsion. These results suggested that lymphatic delivery or tegafur by W/O emulsion was more effective than that by on emulsion due to its differences or formation ability of chylomicrons.

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Study on Direct Teaching Algorithm for Remote Center Motion of Surgical Assistant Robot using Force/Torque Sensor (힘/토크 센서를 이용한 수술보조로봇의 원격중심운동 직접교시 알고리즘 연구)

  • Kim, Minhyo;Jin, Sangrok
    • The Journal of Korea Robotics Society
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    • v.15 no.4
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    • pp.309-315
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    • 2020
  • This study shows a control strategy that acquires both precision and manipulation sensitivity of remote center motion with manual traction for a surgical assistant robot. Remote center motion is an essential function of a laparoscopic surgical robot. The robot has to keep the position of the insertion port in a three-dimensional space, and general laparoscopic surgery needs 4-DoF (degree-of-freedom) motions such as pan, tilt, spin, and forward/backward. The proposed robot consists of a 6-axis collaborative robot and a 2-DoF end-effector. A 6-axis collaborative robot performs the cone-shaped trajectory with pan and tilt motion of an end-effector maintaining the position of remote center. An end-effector deals with the remaining 2-DoF movement. The most intuitive way a surgeon manipulates a robot is through direct teaching. Since the accuracy of maintaining the remote center position is important, direct teaching is implemented based on position control in this study. A force/torque sensor which is attached to between robot and end-effector estimates the surgeon's intention and generates the command of motion. The predefined remote center position and the pan and tilt angles generated from direct teaching are input as a command for position control. The command generation algorithm determines the direct teaching sensitivity. Required torque for direct teaching and accuracy of remote center motion are analyzed by experiments of panning and tilting motion.

3-D Object Tracking using 3-D Information and Optical Correlator in the Stereo Vision System (스테레오 비젼 시스템에서 3차원정보와 광 상관기를 이용한 3차원 물체추적 방법)

  • 서춘원;이승현;김은수
    • Journal of Broadcast Engineering
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    • v.7 no.3
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    • pp.248-261
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    • 2002
  • In this paper, we proposed a new 3-dimensional(3-D) object-tracking algorithm that can control a stereo camera using a variable window mask supported by which uses ,B-D information and an optical BPEJTC. Hence, three-dimensional information characteristics of a stereo vision system, distance information from the stereo camera to the tracking object. can be easily acquired through the elements of a stereo vision system. and with this information, we can extract an area of the tracking object by varying window masks. This extractive area of the tracking object is used as the next updated reference image. furthermore, by carrying out an optical BPEJTC between a reference image and a stereo input image the coordinates of the tracking objects location can be acquired, and with this value a 3-D object tracking can be accomplished through manipulation of the convergence angie and a pan/tilt of a stereo camera. From the experimental results, the proposed algorithm was found to be able to the execute 3-D object tracking by extracting the area of the target object from an input image that is independent of the background noise in the stereo input image. Moreover a possible implementation of a 3-D tele-working or an adaptive 3-D object tracker, using the proposed algorithm is suggested.

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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The effect of emotional priming on the product perceived usability (정서 점화가 제품의 지각된 사용성에 미치는 영향)

  • Kim, Myung Shik;Kim, Hyo Sun;Han, Kwang-Hee
    • Science of Emotion and Sensibility
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    • v.15 no.4
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    • pp.575-584
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    • 2012
  • Decades of psychological research have shown that emotion brings users various kinds of physical and psychological advantages and disadvantages. Moreover, this also impacts human decision-making. However, in spite of the weight of emotion, combined with HCI, relevant research is still insufficient. We hypothesized that the user's temporal emotion could influence the product evaluation, especially in terms of product usability. Two studies were carried out to investigate the effect of induced priming on user evaluations. In exp1, we manipulated participants' temporal emotions using positive and negative images from IAPS. In our results, we saw image priming had a statistically significant effect, with the positive condition group giving the product high ratings for usability. In exp2, emotional image manipulation was conducted with valence and arousal. As a result, we found that the variables of valence and arousal had some interaction effects. These studies have demonstrated that temporally induced emotion could affect users' emotion in different ways, in addition to influencing product evaluations.

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Parthenogenetic Activation of Black Bengal Goat Oocytes

  • Haque, Aminul;Bhuiyan, Mohammad Musharraf Uddin;Khatun, Momena;Shamsuddin, Mohammed
    • Journal of Embryo Transfer
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    • v.26 no.2
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    • pp.123-128
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    • 2011
  • In vitro maturation and activation of oocytes are primary steps towards biotechnological manipulation in embryology. The objectives of the present study were to determine the oocyte recovery rate per ovary, in vitro maturation rates of oocytes and rates of parthenogenetically activation of matured oocytes in Black Bengal goats. All visible follicles were aspirated to recover follicular fluid from individual ovaries (number of ovaries = 456). The immature cumulus oocyte complexes (COCs; n = 1289) were cultured in tissue culture medium (TCM)-199 supplemented with 10% (v/v) fetal bovine serum (FBS) for 27 hours at $39^{\circ}C$ with 5% $CO_2$ in humidified air. The matured oocytes (n = 248) were activated with 5 ${\mu}M$ ionomycin for 5 minutes followed by treatment with 2 mM 6-dimethylaminopurine (6-DMAP) for 4 hours. After activation, oocytes were cultured for another 14 hours in TCM-199 supplemented with bovine serum albumin (BSA) at $39^{\circ}C$ with 5% $CO_2$ in humidified air. The pronucleus formation in activated oocytes was determined by staining with 1% orcein (whole mount technique). Matured oocytes (n = 176) without activation stimuli were used as control. The mean number of oocytes recovered per ovary was $3.5{\pm}0.5$. The proportion of oocytes matured in vitro, confirmed by the presence of first polar body, was $42.1{\pm}4.7%$. Parthenogenetic activation, evidenced by formation of pronucleus, occurred in $37.2{\pm}15.8%$ of matured oocytes. No pronucleus formation was observed in control oocytes. In conclusion, a combination of ionomycin and 6-DMAP induces activation in one third of Black Bengal goats' oocytes.

The Development of Bridge Management System (교량 유지관리 시스템 개발)

  • 이재기;최석근;이현직;박경열
    • Journal of the Korean Society of Surveying, Geodesy, Photogrammetry and Cartography
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    • v.14 no.1
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    • pp.59-68
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    • 1996
  • This study suggests that the management of bridge was developed data which supports to execute analysis a-bout bridge. Basically, it makes standardization that the item of management of bridge keeps the effective data management which demonstrates the designed database by graphic processing technic and development of pro-gram in database manipulation. As a result of this study, through the analysis of the management of bridge, we tried to make the acceptable management item standard and to design database. For the purpose of it, computer program and database design with analyzing current works, stable inspection processing demanding the management of bridge and taking adventage of them. In this study, we can get the minimuim cost of the management of bridge and keep maximum common usage. also we contribute to perform effective data management and management system of bridge.

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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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Transformation of an Alkalin Protease Overproducer, Vibrio metschnikovii Strain RH530, and Improvement of Plasmid Stability by the par Locus

  • Chung, So-Sun;Shin, Yong-Uk;Kim, Hee-Jin;JIn, Chee-Hong;Lee, Hyune-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.11 no.2
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    • pp.222-228
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    • 2001
  • Vibrio metschnikovii strain RH530 is a non-pathogenic, industrially-important alkaline protease producer which has been isolated from wastewater. In this paper, we report on the transformation of this strain by using the method of electroporation. A field strength of $7.5\;kVcm^{-1}$ and $25\;{\mu}F$, and using a 0.2-cm cuvette, appeared to be the optimal conditions for electroporation of the cells with the recombinant pSBCm plasmid carrying the vapK alkaline protease gene and the ColE1 replicon. Cells were subjected to osmotic shock in order to remove extracelluar DNase, and adding 200 mM of sucrose to electroporation buffer cells showed an increased transformation efficiency. Maximum efficiency of transformation was obtained at an early exponential growth phase. Using all of the conditions mentioned above, we routinely obtained a transformation efficiency of more than $10^4{({\mu}g\;plasmid\;DNA)}^{-1}$. The stability of the plasmid pSBCm in V. metschnikovii RH530 was 25% after 18h of growth (27 generations) in the medium without antibiotic selection. The insertion of the par locus to the pSBCm increased the stability of the plasmid up to 42% without selective pressure. The increase in plasmid stability was accompanied by the increase in the productivity of alkaline protease in the recombinant V. metschnikovii strain RH530. Determining optimal conditions for the transformation of the industrially-important, nonpathogenic Vibrio strain, and the improvement of plasmid stability by introducing the par locus into the high copy number plasmid vector, will allow the development of procedures involved in the genetic manipulation of this strain, particularly for its use in the production of industrial enzymes such as alkaline protease.

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