• Title/Summary/Keyword: Cassette

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Toxicological Relevance of Transporters

  • Maeng, Han-Joo;Chung, Suk-Jae
    • Toxicological Research
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    • v.23 no.1
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    • pp.1-9
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    • 2007
  • Transporters are membrane proteins that mediate the transfer of substrate across the cellular membrane. In this overview, the characteristics and the toxicological relevance were discussed for various types of transporters. For drug transporters, the overview focused on ATP-binding cassette transporters and solute carrier family 21A/22A member transporters. Except for OCTN transporters and OATP transporters, drug transporters tend to have broad substrate specificity, suggesting drug-drug interaction at the level of transport processes (e.g., interaction between methotrexate and non-steroidal anti-inflammatory agents) is likely. For metal transporters, transporters for zinc, copper and multiple metals were discussed in this overview. These metal transporters have comparatively narrow substrate specificity, except for multiple metal transporters, suggesting that inter-substrate interaction at the level of transport is less likely. In contrast, the expressions of the transporters are often regulated by their substrates, suggesting cellular adaptation mechanism exists for these transporters. The drug-drug interactions in drug transporters and the cellular adaptation mechanisms for metal transporters are likely to lead to alterations in pharmacokinetics and cellular metal homeostasis, which may be linked to the development of toxicity. Therefore, the transporter-mediated alterations may have toxicological relevance.

Genetic Stability of the Integrated Structural Gene of Guamerin in Recombinant Pichia pastoris

  • Lim, Hyung-Kwon;Kim, Kyeong-Yeon;Lee, Kong-Ju;Park, Doo-Hong;Chung, Soo-Il;Jung, Kyung-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.10 no.4
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    • pp.470-475
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    • 2000
  • Genetic chracterstics of the structural gene of guamerin (a novel elastase inhibitor from Korean leech), integrated into the HIS4 locus of chromosomal DNA of Pichia pastoris along with the $\alpha$-factor leader sequence, were investigated. In the selected clone from candidates, two copies of the integration cassette including the structural gene copies of the integration cassette including the structural gene of guamerin were found in the integration site of the chromosomal DNA of P.pastoris. It was demonstrated that the integrated structural gene of guamerin was stable up to about 70 generations in the relay flask culture. Then, a high-cell-density culture could be fulfilled easily by DO-stat fed-batch culture, in which the cell growth and the recombinant guamerin production reached about 250 of OD600nm and 260 mg/l, respectively. Finally, it was revealed that the DNA sequence of the integrated structural gene of guamerin in P. pastoris was maintained correctly in the end of production cells of relay flask culture and high-cell-density culture.

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Identification of Novel Non-Metal Haloperoxidases from the Marine Metagenome

  • Gwon, Hui-Jeong;Teruhiko, Ide;Shigeaki, Harayama;Baik, Sang-Ho
    • Journal of Microbiology and Biotechnology
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    • v.24 no.6
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    • pp.835-842
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    • 2014
  • Haloperoxidase (HPO, E.C.1.11.1.7) is a metal-containing enzyme oxidizing halonium species, which can be used in the synthesis of halogenated organic compounds, for instance in the production of antimicrobial agents, cosmetics, etc., in the presence of halides and $H_2O_2$. To isolate and evaluate a novel non-metal HPO using a culture-independent method, a cassette PCR library was constructed from marine seawater in Japan. We first isolated a novel HPO gene from Pseudomonas putida ATCC11172 by PCR for constructing the chimeric HPO library (HPO11172). HPO11172 showed each single open-reading frame of 828 base pairs coding for 276 amino acids, respectively, and showed 87% similarity with P. putida IF-3 sequences. Approximately 600 transformants screened for chimeric genes between P. putida ATCC11173 and HPO central fragments were able to identify 113 active clones. Among them, we finally isolated 20 novel HPO genes. Sequence analyses of the obtained 20 clones showed higher homology genes with P. putida or Sinorhizobium or Streptomyces strains. Although the HPO A9 clone showed the lowest homology with HPO11172, clones in group B, including CS19, showed a relatively higher homology of 80%, with 70% identy. E. coli cells expressing these HPO chimeric genes were able to successfully bioconvert chlorodimedone with KBr or KCl as substrate.

Improving Protein Production on the Level of Regulation of both Expression and Secretion Pathways in Bacillus subtilis

  • Song, Yafeng;Nikoloff, Jonas M.;Zhan, Dawei
    • Journal of Microbiology and Biotechnology
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    • v.25 no.7
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    • pp.963-977
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    • 2015
  • The well-characterized gram-positive bacterium Bacillus subtilis is an outstanding industrial candidate for protein expression owing to its single membrane and high capacity of secretion, simplifying the downstream processing of secretory proteins. During the last few years, there has been continuous progress in the illustration of secretion mechanisms and application of this robust host in various fields of life science, such as enzyme production, feed additives, and food and pharmaceutical industries. Here, we review the developments of Bacillus subtilis as a highly promising expression system illuminating strong chemical- and temperatureinducible and other types of promoters, strategies for ribosome-binding-site utilization, and the novel approach of signal peptide selection. Furthermore, we outline the main steps of the Sec pathway and the relevant elements as well as their interactions. In addition, we introduce the latest discoveries of Tat-related complex structures and functions and the countless applications of this full-folded protein secretion pathway. This review also lists some of the current understandings of ATP-binding cassette transporters. According to the extensive knowledge on the genetic modification strategies and molecular biology of Bacillus subtilis, we propose some suggestions and strategies for improving the yield of intended productions. We expect this to promote striking future developments in the optimization and application of this bacterium.

Simultaneous NO2 and RSP Measurements Using Filtration Method (필터방식을 이용한 이산화질소 및 호흡성 분진의 동시 측정)

  • Yang, Won-Ho;Im, Sung-Kuk;Kim, Moon-Hyeon
    • Journal of Environmental Health Sciences
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    • v.34 no.2
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    • pp.148-152
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    • 2008
  • Coated filters were developed to replace the glass impinger methods that use reagent solutions. The purpose of this study was to simultaneously measure nitrogen dioxide ($NO_2$) and respirable suspended particles (RSP) by a filtration method with a cyclone connected to a pump. A first pre-filter for RSP and second filter for $NO_2$, which was soaked in a TEA (Triethanolamine) solution, were loaded into a filter cassette with a pump flow rate of 1.7 l/m. After sampling, the TEA soaked filter was removed from the cassette, placed in a large test tube (10 ml), mixed, and allowed time to develop. The absorbance (abs) of the diazo compound of the $NO_2$ and N-(1-Napthylethylenediamine dihydrochloride) in the color reagent was measured at 545 nm on a spectrophotometer. The collection efficiency(%) of $NO_2$ by each 3 filter soaked in TEA solution and used in the cyclone with a pump flow rate 1.7 l/m was $89{\pm}3%$ and the correlation coefficient between the true $NO_2$ concentration and that determined by the TEA soaked filters was 0.993(p<0.001).

An Experimental Study on the Effect of heat exchanger on turbo fan for air conditioning (열교환기가 공기조화용 터보팬의 성능에 미치는 영향에 대한 실험적 연구)

  • Chang Seung-Yong;Nam Leem-Woo;Joo Won-Gu;Cho Kang-Rae
    • Proceedings of the KSME Conference
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    • 2002.08a
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    • pp.615-618
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    • 2002
  • Turbo-fan for ceiling cassette type air conditioner doesn't operate in general volute. It is operated by porous material, heat exchanger. Heat exchanger increases resistance of air conditioning system and disturbs exit-flow of impeller. Therefore it has some influences on impeller capacity. In this study, we want to how that influence of exchanger on impeller capacity for ceiling cassette type air conditioner. To research, we made circular case that didn't have asymmetric part unlike rectangular case. With and without heat exchanger we measured total pressure and static pressure of impeller and three-dimensional rear flow field From the result, a turbo fan , installed in the 35mm back of fan and operated in heat exchanger, experienced $2{\%}{\~}5{\%}$% total pressure loss over all flow rate. With heat exchanger impeller efficiency decrease as flow rate decrease when flow rate coefficient was below 0.18. Especially when flow rate coefficient was below 0.12, there was $20{\%}{\~}30{\%}$ decrease of impeller efficiency.

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A Survey on Quality of Radiographs in Frontal Projection of the Chest (흉부(胸部) 정면(正面)X선사진(線寫眞)의 화질(畵質)에 관한 검사(檢討))

  • Choi, Jong-Hak;Jeon, Man-Jin;Lee, Chang-Yup
    • Journal of radiological science and technology
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    • v.8 no.2
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    • pp.11-14
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    • 1985
  • The author came to analize and get the following problems after researched radiographic quality in order to find out the necessary factors to improve in radiographs of frontal projection of the chest selected at random of the adult patients (1545 male, 1520 female) who had been examined in 4 departments of radiology of the general hospitals in Seoul and Kyungki area. 1. Problems of x-ray film or of radiographic cassette appeared in 2.97% radiographs on account of selection of the film size (except costophrenic sinus) (1.79%), poor screen-film contact (0.85%), light leakage of cassette (0,33%). 2. Problems of patients' positioning or breathing appeared in 16.57% radiographs of all because of overlapping of lung apex and clavicle (6.98%), overlapping of scapula and lung field (5.87%), asymmetrical projection of clavicles (1.76%), errors in positioning and breathing of the patient (1.96%). 3. Problems of x-ray exposure factors or film processing appeared in 22.25% radiographs because of over-density (2.64%), under-density (3.95%), fog (0.59%), demonstration density under clavicles or lung marking unsharply (3.82%), not clear of lung marking from breast region (0.94%), not clear the lung marking from the part overlapped of heart and lungs (3.92%), not clear the lung marking from the part overlapped of liver and lung (6.49%).

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Mammary Gland-Specific Expression of Biologically Active Human Osteoprotegerin in Transgenic Mice

  • Sung, Yoon-Young;Lee, Chul-Sang
    • Development and Reproduction
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    • v.17 no.1
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    • pp.1-8
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    • 2013
  • Osteoprotegerin (OPG) is a secreted glycoprotein that regulates bone resorption by inhibiting differentiation and activation of osteoclast, thereby potentially useful for the treatment of many bone diseases associated with increased bone loss. In this study, we designed a novel cDNA expression cassette by modifying the potent and mammary gland-specific goat ${\beta}$-casein/hGH hybrid gene construct and examined human OPG (hOPG) cDNA expression in transgenic mice. Six transgenic mice all successfully expressed hOPG in their milk at the level of 0.06-2,000 ${\mu}g/ml$. An estimated molecular weight of the milk hOPG was 55 kDa in SDS-PAGE, which is the same as a naturally glycosylated monomer. This hOPG expression was highly specific to the mammary glands of transgenic mice. hOPG mRNA was not detected in any organs analyzed except mammary gland. Functional integrity of milk hOPG was evaluated by TRAP (tartrate-resistant acid phosphatase) activity assay in bone marrow cell cultures. OPG ligand (OPG-L) treatment increased TRAP activity by two fold but it was completely abolished by co-treatment with transgenic milk containing hOPG. Taken together, our novel cDNA expression cassette could direct an efficient expression of biologically active hOPG, a potential candidate pharmaceutical for bone diseases, only in the mammary gland of transgenic mice.