• Title/Summary/Keyword: laboratory activities

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Target Identification for Metabolic Engineering: Incorporation of Metabolome and Transcriptome Strategies to Better Understand Metabolic Fluxes

  • Lindley, Nic
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2004.06a
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    • pp.60-61
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    • 2004
  • Metabolic engineering is now a well established discipline, used extensively to determine and execute rational strategies of strain development to improve the performance of micro-organisms employed in industrial fermentations. The basic principle of this approach is that performance of the microbial catalyst should be adequately characterised metabolically so as to clearlyidentify the metabolic network constraints, thereby identifying the most probable targets for genetic engineering and the extent to which improvements can be realistically achieved. In order to harness correctly this potential, it is clear that the physiological analysis of each strain studied needs to be undertaken under conditions as close as possible to the physico-chemical environment in which the strain evolves within the full-scale process. Furthermore, this analysis needs to be undertaken throughoutthe entire fermentation so as to take into account the changing environment in an essentially dynamic situation in which metabolic stress is accentuated by the microbial activity itself, leading to increasingly important stress response at a metabolic level. All too often these industrial fermentation constraints are overlooked, leading to identification of targets whose validity within the industrial context is at best limited. Thus the conceptual error is linked to experimental design rather than inadequate methodology. New tools are becoming available which open up new possibilities in metabolic engineering and the characterisation of complex metabolic networks. Traditionally metabolic analysis was targeted towards pre-identified genes and their corresponding enzymatic activities within pre-selected metabolic pathways. Those pathways not included at the onset were intrinsically removed from the network giving a fundamentally localised vision of pathway functionality. New tools from genome research extend this reductive approach so as to include the global characteristics of a given biological model which can now be seen as an integrated functional unit rather than a specific sub-group of biochemical reactions, thereby facilitating the resolution of complexnetworks whose exact composition cannot be estimated at the onset. This global overview of whole cell physiology enables new targets to be identified which would classically not have been suspected previously. Of course, as with all powerful analytical tools, post-genomic technology must be used carefully so as to avoid expensive errors. This is not always the case and the data obtained need to be examined carefully to avoid embarking on the study of artefacts due to poor understanding of cell biology. These basic developments and the underlying concepts will be illustrated with examples from the author's laboratory concerning the industrial production of commodity chemicals using a number of industrially important bacteria. The different levels of possibleinvestigation and the extent to which the data can be extrapolated will be highlighted together with the extent to which realistic yield targets can be attained. Genetic engineering strategies and the performance of the resulting strains will be examined within the context of the prevailing experimental conditions encountered in the industrial fermentor. Examples used will include the production of amino acids, vitamins and polysaccharides. In each case metabolic constraints can be identified and the extent to which performance can be enhanced predicted

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Effect of Extracellular $Ca^{2+}$ and $Ca^{2+}$-ATPase on the Acrosome Reaction of Spermatozoa (세포외 $Ca^{2+}$$Ca^{2+}$-ATPase가 정자의 첨체반응에 미치는 영향)

  • Yung-Keun Oh;Jae-Ho Chang;In-Ho Choi;Noh-Pal Jung;Hyung-Cheul Shin;Byoung-Ju Kwak
    • Biomedical Science Letters
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    • v.4 no.1
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    • pp.27-33
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    • 1998
  • This study has been designed in order to examine a physiological role of $Ca^{2+}$ which has been known as an essential factor for capacitation, to confirm whether the enzyme activity of $Ca^{2+}$-ATPase on capacitation is important or not, and to clarify relationship between various levels of the $Ca^{2+}$ concentration and $Ca^{2+}$-ATPase which has been known to be an important factor of the plasma membranes. In the present study applying quercetin, a $Ca^{2+}$-ATPase inhibitor, the enzymatic effect of $Ca^{2+}$-ATPase on capacitation was found to be remarkable: a significant increase of the transition from the original type (type A) to the type B and the type AR of the spermatozoa. This finding suggests that $Ca^{2+}$-ATPase plays an important role in the efflux and the influx of the $Ca^{2+}$ which has been known to be an essential factor the capacitation and acrosome reaction, and that the inhibitory action of the $Ca^{2+}$-ATPase might be a prerequsite step toward the acrosome reaction. The conclusion reached can be deduced as follows: increment of the intracelluar $Ca^{2+}$ concentration occurred by controlling the slope of $Ca^{2+}$ concentration through $Ca^{2+}$-ATPase activities in both the intra- and extracelluar fluid may be an important procedure for capacitation and acrosome reaction, and ultimately for fertilization of the spermatozoa and the ova.

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Effects of Shiryung-tang Extract on the Liver Injury induced by Ethanol in Rats (시령탕(柴苓湯)이 에탄올 투여로 유발된 흰쥐의 간손상에 미치는 방어효과)

  • Kim, Bum Hoi;Choi, Yung Hyun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.27 no.5
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    • pp.611-616
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    • 2013
  • Alcoholic liver disease (ALD) is a major cause of morbidity and mortality around the world. Although much progress has been made in understanding the pathogenesis of ALD, there remains no effective therapy for it. Accumulated evidence indicates that oxidative stress is the main pathological factors in the development of ALD. Ethanol administration causes accumulation of reactive oxygen species (ROS), including superoxide, hydroxyl radical, and hydrogen peroxide. ROS, in turn, cause lipid peroxidation of cellular membranes, and protein and DNA oxidation, which results in hepatocyte injury. In addition to pro-oxidants formation, antioxidants depletion caused by ethanol administration also results in oxidative stress. The objective of this study is to investigate the effects of Shiryung-tang extract on the chronic alcoholic liver injury induced by EtOH. Male Sprague Dawley rats were used in this study. All rats were maintained under standard laboratory conditions ($23{\pm}1^{\circ}C$, 12h light/12h dark cycles). All animals (n=30) were randomly divided into following groups: (1) Normal group, treated with distilled water (n=10); (2) Control group, treated with ethanol (n=10); (3) Sample group, treated with ethanol + pharmacopuncture (n=10). For oral administration of ethanol in Control and Sample group, the ethanol was dissolved in distilled water in concentrations of 25%(v/v). Throughout the experiment of 8 week, the rats were allowed free access to water and standard chow. Sample group were administrated by Shiryung-tang extract daily for 8 weeks. Control group were given normal saline for same weeks. As a results, the oral administration of ethanol for 8 weeks leads to hepatotoxicity. The levels of hepatic marker such as HDL-cholesterol, triglyceride, aspartate aminotransferase and alanine aminotransferase were altered. The ethanol also increased lipid peroxidation and depletion of antioxidant enzyme activities as well as hepatic tissue injury. However, the treatment of Shiryung-tang extract prevented all the alterations induced by ethanol and returned their levels to near normal. These data suggest that Shiryung-tang extract could have a beneficial effect in inhibiting the oxidative damage induced by chronic ethanol administration. Therefore, Shiryung-tang extract can be a candidate to protect against EtOH-induced liver injury.

Changes of some chemical constituents in different soil depth with textures of Fluvio-marine soil under assessment of reclamation duration (간척년수(干拓年數)에 따른 토성(土性) 및 작토층위별(作土層位別) 수종(數種) 화학성분변화(化學成分變化) 차이(差異)에 관(關)한 연구(硏究))

  • Kim, Seong Chae
    • Korean Journal of Soil Science and Fertilizer
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    • v.20 no.1
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    • pp.23-28
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    • 1987
  • A series of field and laboratory experiments were conducted to find out the changes of some soil chemical constituents in different soil depth with three different soil textures as Bongnam-clayey, Gwanghwal-silty loam and Mangyeong fine sandy soil on the assessment of reclamation duration in Fluvio-marine soil. The result obtained were summarized as follows; 1. Exchangeable potassium, calcium, magnessium, sodium with manganes, silica and cation exchange capacity were remarkably decreased with assessment of reclamation duration. Decreasing tendency was pronounced more in sandy soil than silty loam or clayly soil. 2. Soil organic matter content in top soil was increased with increasing of reclamation years. From reclamation time, about 30 years in clayly and about 80 years in silty loam and sandy soil were necessary for the maintanance of averaged organic matter content to 2.5%. 3. Potassium activity ratio ($K/{\sqrt{Ca+Mg+Na}}$) was differed with soil depth, soil texture and assessment of reclamation duration. About 50 years is may be necessary for the maintanance of potassium activities ratio in ordinary paddy soil of 0.05-0.2. 4. Percent of adsorbed sodium (PAS), ratio of adsorbed sodium (RAS) and ratio of sodium adsorbtion(RSA) were remarkably decreased with assessment of reclamation duration. Specially, decreasing tendency was pronounced more in sandy soil than silty loam and clayly soil 5. Amount of clay content in subsoil was appearently decreased during 30 years of reclamation, but clay contents in top soil was appearently decreased 50 years after reclamation.

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Microcosm Study for Bioremediation of Oil-Contaminated Pebble Environments (자갈로 구성된 미소환경에서 미생물제제에 의한 유류분해)

  • Sim, Doo-Suep;Sohn, Jae-Hak;Kim, Sang-Jin
    • Korean Journal of Microbiology
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    • v.34 no.3
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    • pp.101-107
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    • 1998
  • Biological treatment of Arabian light crude oil-contaminated pebble was investigated in laboratory microcosms after supplementation with inorganic nutrients and oil-degrading microorganisms. Glass columns ($10cm{\times}20cm$) were used as microcosms and each microcosm was filled with pebbles of diameter less than 40 mm. After initial oil contamination of 2.4% (w/v), Inipol EAP-22 or slow release fertilizer (SRF) was added as inorganic nutrients and microorganisms were sprayed over pebbles. When $C_{17}$/pristane and $C_{18}$/phytane ratios were used as a marker for oil biodegradation, both ratios for microcosm supplemented with SRF and microorganisms were the lowest (below detectable range) after 92 days. Elimination of oil by abiotic processes, however, were minimal with decrease of $C_{17}$/pristane and $C_{18}$/phytane ratios from 3.55 and 2.41 to 3.06 and 1.50, respectively. The numbers of heterotrophic and oil-degrading microorganisms, and biological activity (dehydrogenase activity) corresponded to the course of biodegradation activities in all microcosms. During the whole experimental period, there was no significant nutrient deficiency only in the microcosm with SRF and microorganisms. It seemed that a continuous supply of inorganic nutrients using SRF was the most important factor for the successful performance of biological treatment in oil-contaminated pebbles.

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Chemical Control Effect Against Spot Clothing Wax Cicada, Lycorma delicatula (Hemiptera: Fulgoridae) Nymphs and Adults (꽃매미(Lycorma delicatula) 약충과 성충에 대한 약제방제 효과)

  • Kim, Sun-Kook;Lee, Gi-Yeul;Shin, Yon-Ho;Kim, Gil-Bah
    • The Korean Journal of Pesticide Science
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    • v.14 no.4
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    • pp.440-445
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    • 2010
  • This study was carried out to investigate the toxicities of five registered insecticides to the Lycorma delicatula nymphs and adults. Insecticidal activities were evaluated by testing systemic and residual effects in the laboratory, and control efficacy in the vineyard. For the 3rd nymphal instars and adults, etofenprox+diazinon, chlorpyrifos, etofenprox, dinotefuran and imidacloprid showed perfect insecticidal activity. The systemic effects of dinotefuran and etofenprox+diazinon on grape roots at half concentration were showed the adult mortality of 82.2% and 84.4%, respectively. Chlorpyrifos at recommended concentration was showed mortality of 86.0%. The leaf systemic effects at recommended and half concentrations of all insecticides except dinotefuran were lower than 65% but the mortality at double concentration of chlorpyrifos, dinotefuran, etofenprox were more than 82%. The residual effect between etofenprox+diazinon and dinotefuran at recommended and double concentrations were 100% at 14 days after treatment, the other insecticides have low efficacy. All the four insecticides showed 87% control value on nymphs in the field test and adults appeared more than 96% control value st and adinotefuran, etofenprox+diazinon, however, the other insecticides decreased to 59.1 % and 61.2%. Therefore, dinotefuran and etofenprox+diazinon showing high systemic effects to roots and long residual effects to leafs have high control efficacies.

Studies on chemical components of fermented maeh in the brewing of Maggerley (Korean wine) (막걸리 제조시 술덧의 성분동태에 관한 연구 1)

  • 이성범;장원길;임병종;김덕치
    • Korean Journal of Microbiology
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    • v.7 no.4
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    • pp.153-158
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    • 1969
  • It is the most important thing to substitute sweet potato for wheat flour as the brewing material of Maggerley(Korean wine) in order to save precious wheat flour which is now consumed as the sole material in it. In this study, the fermenting mash material has been prepared with combination of wheat (KIorean wine) as a first step of sabing wheat flour consuming. The combination ratio of wheat flour and sweet potato starch in mashing, in this experiment, are such as mashing plot of 100% wheat flour (No.3 plot), 50% wheat flour plus 50 % of sweet potato starch (No.2plot), 30% wheat flour plus 70% starch syrup of sweet potato (No. 4 plot), and 60% wheat flour plus sweet potato starch (No. 1 plot). The results of fermentation and chemical components of each mash in experimental plot are as following : 1. Alocholic fermentation was most vigorous in No.4 plost among them, the next are in order to hydrolyze and break down the protein sourece to glucose and amino acid, the new enzymic source were added in experimantal amsh before putting yeasts, those enzymic sources have been developed and prepared in this laboratory No.4 and No.2 plot at same degree. Generally, the activities of alcholic fermentation of all plots are vigorous and fair. The acidity of each fermented mash is in the rangw of 3.7-4.8. 2. Amounts of amino acids in fermented mash are in the range of 145 mg per 100 ml and 158 mg for all experiments, and these amounts are much more than those of present commerical Maggerley. However, the amount of it in the No.4 plots is much less among the m, this is due to the lack of protein source in starch syrup. 3. Amounts of organic acids in fermented mash are in the range of 0.44g per 100ml and 0.62g, these amounts are generally much more than those of the present commercial Maggerley. 4. The result of sensory test for the experimental Maggerley shows good taste and flavour in comparison with those of commercial Maggerely. 5. It is proved that these experimental fermentation of Maggerley is succesful in the partial substitution of sweet potato starch potato starch for wheat flour, and it is also be able to make a good Maggerley in all aspects of quality.

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Interministerial GHS Activities and Implementation in Korea

  • Yu, Il-Je
    • Proceedings of the Korean Environmental Health Society Conference
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    • 2005.06a
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    • pp.240-248
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    • 2005
  • To implement a globally harmonized system of classification and labeling of chemicals (GHS) in Korea, an interminsterial GHS working group involving 6 ministries established an expert working group composed of 7 experts from relevant organizations and one private consultant to prepare an officialKorean GHS version by March, 2005. As such, the translation and review of the official Korean GHS version, including annexes, started in October, 2004 and was completed on March 15, 2005. The official Korean GHS version has now been posted on the websites of the relevant ministries and organizations to solicit public opinions. The official Korean GHS version will be finalized after a public hearing scheduled forMay, 2005. Collaborative efforts as regards implementing and disseminating the GHS in Korea will be continued to avoid any confusion or duplication and for effective use of resources. The globally harmonized system of classifying and labeling chemicals (GHS) was originally adopted in 1992 at the United Nations Conference on Environment and Development (UNCED), as subsequently reflected in Agenda 21 chapter 19. The work was coordinated and managed under the auspices of the Interorganization Programme for the Sound Management of Chemicals(IOMC) Coordinating Group for the Harmonization of Chemical Classification Systems (UNCEGHS). The technical focal points for completing the work were the International Labour Organization (ILO); Organization for Economic Cooperation and Development (OECD); and United Nations Economic and Social Council's Subcommittee of Experts on the Transport of Dangerous Goods (UNSCETDG). The work was finalized in October 2002, and the World Summit on Sustainable Development in Johannesburg on 4 September 2002 encouraged countries to implement the new GHS as soon as possible with a view to having the system fully operational by 2008 (UN, 2003). Implementation has already started with pilot countries introducing the system to their national practices in different regions of the world. The GHS text, called the purple book, becameavailable as a W publication in early 2003. The GHS text, called the purple book, becameavailable as a UN publication in early 2003. The GHS system will be kept dynamic, and regularly revised and made more efficient as experience is gained in its implementation. While national or regional governments are the primary audiences for this document, it also contains sufficient context and guidance for those in industry who will ultimately be implementing the national requirements that will be introduced (UN, 2003). The Japanese government published their official Japanese GHS version, the first in Asia, in April 2004 after starting work in January 2003 based on an interministerial chemical coordination committee involving 7 ministries, including the Ministry of Foreign Affairs, Ministry of Internal Affairs and Communications, Ministry of Health, Labour, and Welfare, Ministry of Agriculture, Forestry and Fisheries, Ministry of Economy, Trade and Industry, Ministry of Land, Infrastructure, and Transport, and Ministry of Environment (MOE, 2004). Accordingly, similar to the Japanese GHS efforts, this paper presents the interministerial efforts involved in publishing the official Korean GHS version.

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A Study on the Effect of Aging on the Xylene Toxicity in Rats (흰쥐 성장기간에 따른 Xylene의 독성에 관한 연구)

  • 이혜자;이상희;전태원;이상일;윤종국
    • Biomedical Science Letters
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    • v.6 no.3
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    • pp.193-199
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    • 2000
  • To evaluate an effect of aging on the xylene toxicity, 50% m-xylene in olive oil (0.25 ml/100 g body wt.) was administered to 5 week and 12 week-old rats one times intraperitoneally and sacrificed at 24 hrs afterwards. The increasing rate of urinary m-methylhippuric acid concentration was higher in 12 week-old rats than 5 week-old rats by the treatment of m-xylene. On the liver function findings, i.e., liver weigh/body weight (%), serum levels of ALT activity and hepatic malondialdehyde content, 12 week-old rats showed more severe liver injury than 5 weeks those in xylene-treated rats. And the hepatic cytochrome P-450 contents was higher in 12 weeks rats than those of in 5 week-old rats, but the increasing rate of that was lower in 12 week-old rats. Hepatic alcohol dehydrogenase activity was also higher in 12 week-old rats than in 5 week-old rats whereas the increasing rate of that was higher in 5 week-old than those in 12 week-old rats by the xylene treatment. Furthermore, the hepatic aldehyde dehydrogenase activities were no differences between the 5 and 12 week-old rats both in the control and xylene-treated group. In conclusion, age may influences upon the hepatotoxicity with xylene and it may be responsible for xylene metabolism in rats.

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Neurotoxicity of Synthetic Cannabinoids JWH-081 and JWH-210

  • Cha, Hye Jin;Seong, Yeon-Hee;Song, Min-Ji;Jeong, Ho-Sang;Shin, Jisoon;Yun, Jaesuk;Han, Kyoungmoon;Kim, Young-Hoon;Kang, Hoil;Kim, Hyoung Soo
    • Biomolecules & Therapeutics
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    • v.23 no.6
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    • pp.597-603
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    • 2015
  • Synthetic cannabinoids JWH-018 and JWH-250 in 'herbal incense' also called 'spice' were first introduced in many countries. Numerous synthetic cannabinoids with similar chemical structures emerged simultaneously and suddenly. Currently there are not sufficient data on their adverse effects including neurotoxicity. There are only anecdotal reports that suggest their toxicity. In the present study, we evaluated the neurotoxicity of two synthetic cannabinoids (JWH-081 and JWH-210) through observation of various behavioral changes and analysis of histopathological changes using experimental mice with various doses (0.1, 1, 5 mg/kg). In functional observation battery (FOB) test, animals treated with 5 mg/kg of JWH-081 or JWH-210 showed traction and tremor. Their locomotor activities and rotarod retention time were significantly (p<0.05) decreased. However, no significant change was observed in learning or memory function. In histopathological analysis, neural cells of the animals treated with the high dose (5 mg/kg) of JWH-081 or JWH-210 showed distorted nuclei and nucleus membranes in the core shell of nucleus accumbens, suggesting neurotoxicity. Our results suggest that JWH-081 and JWH-210 may be neurotoxic substances through changing neuronal cell damages, especially in the core shell part of nucleus accumbens. To confirm our findings, further studies are needed in the future.