• Title/Summary/Keyword: host manipulation

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Production of Therapeutic Glycoproteins throgh the Engineering of Glycosylation Pathway in Yeast

  • Roy, Samir-Kumar;Yasunori Chiba;Yoshifumi Jigami
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.4
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    • pp.219-226
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    • 2000
  • The application of recombinant DNA technology to restructure metabolic net-work can change metabolite and protein products by altering the biosynthetic pathways in an organism. Although some success has been achieved, a more detailed and thorough investigation of this approach is certainly warranted since it is clear that such methods hold great potential based on the encouraging results obtained so far. In last decade, there have been tremendous advances in the field of glycobiology and the stage has been set for the biotechnological production of glycoproteins for therapeutic use. Today glycoproteins are one of the most important groups of pharmaceutical products. In this study the attempt was made to focus on identifying technologies that may have general application for modifying glycosylation pathway of the yeast cells in order to produce glycoproteins of therapeutic use. The carbohydrates of therapeutic recombinant glycoproteins play very important roles in determining their pharmacokinetic properties. A number of biological interactions and biological functions mediated by glycans are also being targeted for therapeutic manipulation in vivo. For a commercially viable production of therapeutic glycoproteins a metabolic engineering of a host cell is yet to be established.

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The role of rumen microbiota in enteric methane mitigation for sustainable ruminant production

  • Takumi Shinkai;Shuhei Takizawa;Miho Fujimori;Makoto Mitsumori
    • Animal Bioscience
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    • v.37 no.2_spc
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    • pp.360-369
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    • 2024
  • Ruminal methane production functions as the main sink for metabolic hydrogen generated through rumen fermentation and is recognized as a considerable source of greenhouse gas emissions. Methane production is a complex trait affected by dry matter intake, feed composition, rumen microbiota and their fermentation, lactation stage, host genetics, and environmental factors. Various mitigation approaches have been proposed. Because individual ruminants exhibit different methane conversion efficiencies, the microbial characteristics of low-methane-emitting animals can be essential for successful rumen manipulation and environment-friendly methane mitigation. Several bacterial species, including Sharpea, uncharacterized Succinivibrionaceae, and certain Prevotella phylotypes have been listed as key players in low-methane-emitting sheep and cows. The functional characteristics of the unclassified bacteria remain unclear, as they are yet to be cultured. Here, we review ruminal methane production and mitigation strategies, focusing on rumen fermentation and the functional role of rumen microbiota, and describe the phylogenetic and physiological characteristics of a novel Prevotella species recently isolated from low methane-emitting and high propionate-producing cows. This review may help to provide a better understanding of the ruminal digestion process and rumen function to identify holistic and environmentally friendly methane mitigation approaches for sustainable ruminant production.

A clinical study of the effect of Nd:YAG laser irradiation on inflamed gingiva (염증성 치은에 대한 Nd:YAG laser 조사효과에 관한 임상적 연구)

  • Cho, Hyung-Soo;Kim, Hyun-Sup;Lim, Kee-Jung;Kim, Byung-Ock;Han, Kyung-Yoon
    • Journal of Periodontal and Implant Science
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    • v.26 no.2
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    • pp.531-541
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    • 1996
  • Periodontal disease is characterized by destruction of supporting tissues caused by invasion of plaque bacteria and defense mechanism of host. Many dentists are very interested in laser therapy on various intraoral soft tissue lesions including inflammatory periodontal pocket. In order to determine the therapeutic effect of intrapocket irradiation of a pulsed- Nd : YAG laser on the inflammatory periodontal pockets, bilateral 60 teeth with 4-6mm in probing pocket depth and gingival inflammation were selected and evaluated by sulcus bleeding index(SBI), and plaque index(pI) for baseline record. Intrapocket irradiation($300{\mu}m$ fiber optic, I.5W power, for 2 min.) of a pulsed-Nd : YAG laser(EL.EN.EN060, Italy) was applied on half of them. As the control group, the same procedure except power-off was repeated on the contralateral 30 teeth. At 1-, 2-, 3-, and 4-week after intrapocket manipulation, every tooth was reevaluated by the same clinical indices. And the difference between the lased group and control group was statistically analyzed by paired t-test and Chi-square test in Microstat program. Following results were obtained: 1. Until I-week and 2-week after intrapocket manipulation, SBI was lowered in both lased group and control group, compared to baseline SBI, but from 3-week after, the recovering tendency toward baseline was noted, and at only 2-week after, the number of teeth showing lowered SBI was significantly more in lased group than in control group(p<0.05). 2. PI of both lased group and control group was lowered through whole experimental period from I-week to 4-week after, compared to baseline PI(p<0.05), but there was no significant difference between lased group and control group(p>0.1). The results suggest that intrapocket irradiation of a pulsed-Nd:YAG laser may lead somewhat remission of gingival inflammation, but for more favorable therapeutic result the thorough root planing should be necessarily accompanied with gingival curettage.

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Stability of pUC-Derived Plasmids with a Fluorescence Marker in Pectobacterium carotovorum subsp. carotovorum and subsp. betavasculorum

  • Hur, Woon-Yung;Roh, Eun-Jung;Oh, Chang-Sik;Han, Man-Wi;Lee, Seung-Don;Kim, Doo-Ho;Heu, Sung-Gi
    • The Plant Pathology Journal
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    • v.25 no.3
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    • pp.286-290
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    • 2009
  • The stability of three different kinds of pUC-derived plasmids, pDsRed, pZsYellow, and pGFPuv, was investigated in Pectobacterium strains to utilize those plasmids as tracers. All three plasmids pDsRed, pZsYellow and pGFPuv showed their specific colors in Pectobacterium strains. Especially, the plasmid pDsRed conferred bright pink colonies on the Pectobacterium strains. When the bacteria lost the plasmid pDsRed, the colonies turned white, suggesting that the plasmid could be a good marker system for Pectobacterium strains on different environmental conditions. The effect of the antibiotic pressure on the stability of the plasmid was different depending on the host bacteria. P. carotovorum subsp. betavasculorum was more sensitive to the antibiotic pressure than P. carotovorum subsp. carotovorum Pcc21. However, temperature change significantly affected plasmid stability on both Pectobacterium strains. Almost all strains lost the plasmids with the shift in temperature from $28^{\circ}C$ to $37^{\circ}C$. Presence of the plasmids did not affect bacterial pathogenicity on their own host plants. Among three plasmids, pZsYellow was not useful as a marker because the yellow fluorescent proteins from pZs Yellow were interfered with the yellow natural fluorescence of the plant tissues induced by the defense system. Since the red color of DsRed can be seen with naked eyes, plasmid pDsRed was applicable as a marker. However, the color change was slow so that additional manipulation to increase the expression speed was necessary. Plasmid pGFPuv could serve as a perfect marker without any problem, tracing the reproduction and spread of the plant pathogens perfectly.

Recent Progress in Biotechnology-based Gene Manipulating Systems to Produce Knock-In/Out Mouse Models

  • Lee, Woon Kyu;Park, Joong Jean;Cha, Seok Ho;Yun, Cheol-Heui
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.5
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    • pp.745-753
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    • 2008
  • Gene-manipulated mice were discovered for the first time about a quarter century ago. Since then, numerous sophisticated technologies have been developed and applied to answer key questions about the fundamental roles of the genes of interest. Functional genomics can be characterized into gain-of-function and loss-of-function, which are called transgenic and knock-out studies, respectively. To make transgenic mice, the most widely used technique is the microinjection of transgene-containing vectors into the embryonic pronucleus. However, there are critical drawbacks: namely position effects, integration of unknown copies of a foreign gene, and instability of the foreign DNA within the host genome. To overcome these problems, the ROSA26 locus was used for the knock-in site of a transgene. Usage of this locus is discussed for the gain of function study as well as for several brilliant approaches such as conditional/inducible transgenic system, reproducible/inducible knockdown system, specific cell ablation by Cre-mediated expression of DTA, Cre-ERTM mice as a useful tool for temporal gene regulation, MORE mice as a germ line delete and site specific recombinase system. Techniques to make null mutant mice include complicated steps: vector design and construction, colony selection of embryonic stem (ES) cells, production of chimera mice, confirmation of germ line transmission, and so forth. It is tedious and labor intensive work and difficult to approach. Thus, it is not readily accessible by most researchers. In order to overcome such limitations, technical breakthroughs such as reporter knock-in and gene knock-out system, production of homozygous mutant ES cells from a single targeting vector, and production of mutant mice from tetraploid embryos are developed. With these upcoming progresses, it is important to consider how we could develop these systems further and expand to other animal models such as pigs and monkeys that have more physiological similarities to humans.

Expression of Matrix Metalloproteinases (MMPs) and Their Tissue Inhibitors (TIMPs) in Frozen Sperm of Rabbit (동결융해 후 토끼 정자의 Matrix Metalloproteinases (MMPs)와 Their Tissue Inhibitors (TIMPs) 발현 양상)

  • Kim, Sang Hwan;Choi, Hwa Sik;Yoon, Jong Taek
    • Journal of Animal Reproduction and Biotechnology
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    • v.34 no.3
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    • pp.247-252
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    • 2019
  • We observed MMPs expression in all sperm groups, with pro-MMP showing lower expression than active MMPs. According to the results from each freezing extender, the sperm membrane integrity (HOST: Hypoosmotic Swelling Test) analysis in TCGGD (Tris 250 mM, Citric acid 88 mM, Glucose 47 mM, Glycerol 3%, Dimethylsulpoxide 3.5 M) is 59.8 ± 0.7, TCGSD (Tris 250 mM, Citric acid 88 mM, Glucose 47 mM, Sucrose 0.1 M, Dimethylsulpoxide 3.5 M) is 59.3 ± 0.5 were significantly higher (p < 0.05) among the experimental groups. And MMPs analysis result, we observed MMPs expression in all sperm groups, with pro-MMP showing lower expression than active MMPs. The expression of active MMP-2 was the highest in sperms frozen in TCGSD and TCGD (Tris 250 mM, Citric acid 88 mM, Glucose 47 mM, Dimethylsulpoxide 3.5 M), Meanwhile, sperms from the TCGGD and TCGED (Tris 250 mM, Citric acid 88 mM, Glucose 47 mM, Ethylene glycol 3%, Dimethylsulpoxide 3.5 M) group showed lower level of active MMP-2 expression. Together, these results indicate that adding glycerol or sucrose to the sperm freezing buffer would not only suppress MMPs expression but also minimize DNA fragmentation, providing a mean to improve the success rate in the in vitro manipulation of rabbit sperms. Therefore, these results suggest that TCGGD or TCGSD extender method for freezing-thawing of rabbit sperm increased the viability after thawing.

Recent Advances in Gut Microbiology and Their Possible Contribution to Animal Health and Production - A Review -

  • Kobayashi, Yasuo;Koike, Satoshi;Taguchi, Hidenori;Itabashi, Hisao;Kam, Dong K.;Ha, Jong K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.6
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    • pp.877-884
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    • 2004
  • Although gut microbial functions have been analyzed through cultivation of isolated microbes, molecular analysis without cultivation is becoming a popular approach in recent years. Gene cloning studies have partially revealed the mechanisms involved in fiber digestion of individual microbe. The molecular approach finally made it possible to analyze full genomes of the representative rumen cellulolytic bacteria Fibrobacter and Ruminococcus. The coming database may contain useful information such as regulation of gene expression relating to fiber digestion. Meanwhile, unculturable bacteria are still poorly characterized, even though they are main constituents of gut microbial ecosystem. The molecular analysis is essential to initiating the studies on these unculturable bacteria. The studies dealing with rumen and large intestine are revealing considerable complexity of the microbial ecosystems with many undescribed bacteria. These bacteria are being highlighted as possibly functional members contributing to feed digestion. Manipulation of gut bacteria and gut ecology for improving animal production is still at challenging stage. Bacteria newly introduced in the rumen, whether they are genetically modified or not, suffer from poor survival. In one of these attempts, Butyrivibrio fibrisolvens expressing a foreign dehalogenase was successfully established in sheep rumen to prevent fluoroacetate poisoning. This expands choice of forages in tropics, since many tropic plants are known to contain the toxic fluoroacetate. This example may promise the possible application of molecular breeding of gut bacteria to the host animals with significance in their health and nutrition. When inoculation strategies for such foreign bacteria are considered, it is obvious that we should have more detailed information of the gut microbial ecology.

Research about Security Attack Methods to Arduino Boards Using Temporary Files Data Manipulation (임시파일 데이터 조작을 통한 아두이노 보드 공격 기법에 관한 연구)

  • Lee, Woo Ho;Jung, Hyun Mi;Jeong, Kimoon
    • Journal of the Korea Convergence Society
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    • v.8 no.11
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    • pp.21-27
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    • 2017
  • Internet of Things(IoT), which is developing for the hyper connection society, is based on OSHW (Open Source Hardware) such as Arduino and various small products are emerging. Because of the limitation of low performance and low memory, the IoT is causing serious information security problem that it is difficult to apply strong security technology. In this paper, we analyze the vulnerability that can occur as a result of compiling and loading the application program of Arduino on the host computer. And we propose a new attack method that allows an attacker to arbitrarily change the value input from the sensor of the arduino board. Such as a proposed attack method may cause the arduino board to misinterpret environmental information and render it inoperable. By understanding these attack techniques, it is possible to consider how to build a secure development environment and cope with these attacks.

The software architecture for the internal data processing in Gigabit IP Router (기가비트 라우터 시스템에서의 내부 데이터 처리를 위한 소프트웨어 구조)

  • Lee, Wang-Bong;Chung, Young-Sik;Kim, Tae-Il;Bang, Young-Cheol
    • The KIPS Transactions:PartC
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    • v.10C no.1
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    • pp.71-76
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    • 2003
  • Internet traffic is getting tremendously heavier due to the exponential growth of the Internet users, the spread of the E-commerce and the network games. High-speed routers for fast packet forwarding are commercially available to satisfy the growing bandwidth. A high-speed router, which has the decentralized multiprocessing architecture for IP and routing functions, consists of host processors, line interfaces and switch fabrics. In this paper, we propose a software architecture tuned for high-speed non-forwarding packet manipulation. IPCMP (Inter-Processor Communication Message Protocol), which is a mechanism for IPC (Inter-Processor Communication), is also proposed and implemented as well. Proposed IPC mechanism results in faster packet-processing rate by 10% as compared to the conventional IPC mechanism using UDP/IP.

Application of Mycorrhizal Research to Agriculture and Forestry (균근연구(菌根硏究)의 농림업(農林業)에의 응용(應用))

  • Lee, Kyung Joon;Lee, Don Koo;Lee, Won Kyu;Koo, Chang Duck
    • Journal of Korean Society of Forest Science
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    • v.59 no.1
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    • pp.121-142
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    • 1983
  • Recently mycorrhizal research has been one of the most fast-growing research areas in modern plant science and microbiology. The application potential of mycorrhizal techniques to agriculture and forestry is enormous in view of the ubiquitous nature of mycorrhizae and known benefits of mycorrhizae to host plants. Unfortunately, very few scientists in Korea are currently involved in mycorrhizal research. When a team of American plant pathologists visited Korea in September 1982 to participate in the Korea-U.S.A. Joint Seminar on Forest Diseases and Insect Pests, they were surprised by the principal author's statement that there was no single research project on mycorrhizae sponsored by Korean government or any scientific institutions. The author initiated a few years ago a research project on the ecology of tree mycorrhizae with a foreign financial support. Major areas of interest were survey of ectomycorrhizae in relation to soil fertility, taxonomic distribution of mycorrhizae among woody plants, identification of ectomycorrhizal fungi, and growth response of woody plants to artificial inoculation. In spite of the enormous application potential of mycorrhizae to agronomic plants, the subject of mycorrhizae has not been recognized by Korean agronomists, foresters or pathologists. The purpose of this review rather written in Korean is to introduce the techniques of mycorrhizal research to Korean scientists and to urge them to participate in challenging new scientific field which might bring us a remarkable increase in crop productivity and tree growth through manipulation of this unique symbiosis. In this review, following topics were discussed in the same order: introduction; brief history of mycorrhizal research; morphology and classification of mycorrhizae; distribution of mycorrhizae in plant kingdom and in soil profile; physiology of mycorrhizae (functions, mineral nutrition, mycorrhizal formation); interaction of mycorrhizae with soil-born plant pathogens. mycorrhizae in nitrogen-fixing plants; application of mycorrhizal techniques to nursery practices (isolation, culture, inoculation, and response); prospect in the future.

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