• Title/Summary/Keyword: PGH

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Expression of the Recombinant Porcine GH Gene In Vitro Using Tetracycline Inducible Expression System (In Vitro에서의 Tetracycline Inducible Expression System에 의한 재조합 돼지 성장호르몬 유전자의 발현)

  • Kwon Mo Sun;Koo Bon Chul;Kim Teoan
    • Reproductive and Developmental Biology
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    • v.29 no.1
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    • pp.49-55
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    • 2005
  • We cloned cDNA of the PGH(porcine growth hormone) gene and constructed retrovirus vector designed to express PGH gene under the regulation of CMV (cytomegalovirus) promoter. To maximize the expression, WPRE(woodchuck hepatitis virus posttranscriptional regulatory element) sequence was placed at the downstream of the PGH gene. After infection with recombinant viruses, approximately 1×10/sup 6/ PFF(porcine fetal fibroblast) cells released PGH protein into the media as much as 1,400 ng. In a subsequent experiment, a modifications of the retrovirus vector was made to express the PGH gene in a teracycline-inducible manner. In PFF cells carrying these viral vector sequences, addition of doxycycline to the media resulted in 2∼6 fold increase in PGH synthesis. In the modified retrovirus vectors, the WPRE sequence also played a role in boosting the effect of the tetracycline induction. This result indicates that our tetracycline-inducible expression system might be a promising candidate in alleviating the complicate physiological problems caused by constitutive expression of the exogenous genes in the transgenic animals.

Comparison study of dermal cell toxicity and zebrafish brain toxicity by humidifier sterilizer chemicals (PHMG, PGH, CMIT/MIT) (가습기 살균제 성분(PHMG, PGH, CMIT/MIT)의 사람 피부세포 독성 및 제브라피쉬 뇌신경 독성 비교 연구)

  • Cho, Kyung-Hyun;Kim, Jae-Ryong
    • Korean Journal of Environmental Biology
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    • v.38 no.2
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    • pp.271-277
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    • 2020
  • Toxicities to many organs caused by humidifier disinfectants have been reported. Recently, humidifier disinfectants have been reported to cause cardiovascular, embryonic, and hepatic toxicities. This study was designed to investigate the toxic mechanism of humidifier disinfectants and compare toxicity in a cellular model and a zebrafish animal model. Because brain toxicity and skin toxicity have been less studied than other organs, we evaluated toxicity in a human dermal cell line and zebrafish under various concentrations of humidifier disinfectants that included polyhexamethyleneguanidine phosphate (PHMG), oligo-[2-(2-ethoxy)-ethoxyethyl-guanidinium-chloride] (PGH) and methylchloroisothiazolinone/methylisothiazolinone (CMIT/MIT). A human dermal fibroblast cell line was treated with disinfectants (0, 2, 4, 6, 8, and 16 mg L-1) to compare their cytotoxicity. The fewest PHMG-treated cells survived (up to 33%), while 49% and 40% of the PGH- and CMIT/MIT-treated cells, respectively, survived. The quantification of oxidized species in the media revealed that the PHMG-treated cells had the highest MDA content of around 28 nM, while the PGH- and CMIT/MIT-treated cells had 13 and 21 nM MDA, respectively. As for brain toxicity, treatment of the zebrafish tank water with CMIT/MIT (final 40 mg L-1) for 30 min resulted in a 17-fold higher production of reactive oxygen species (ROS) than in the control. Treatment with PGH or PHMG (final 40 mg L-1) resulted in 15- and 11-fold higher production, respectively. The humidifier disinfectants (PHMG, PGH, and CMIT/MIT) showed severe dermal cell toxicity and brain toxicity. These toxicities may be relevant factors in understanding why some children have language disorders, motor delays, and developmental delays from exposure to humidifier disinfectants.

Development of a Coupled Enzyme Assay Method for Microsomal Prostaglandin E Synthase Activity

  • Choi, Kyung-A;Park, Sung-Jun;Yu, Yeon-Gyu
    • Bulletin of the Korean Chemical Society
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    • v.31 no.2
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    • pp.384-388
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    • 2010
  • Human microsomal prostaglandin E synthase-1 (mPGES-1) catalyzes the conversion of prostaglandin $H_2$ ($PGH_2$) into prostaglandin $E_2$ ($PGE_2$). To establish a stable and efficient method to assess the activity of mPGES-1, a coupled enzyme assay system using mPGES-1, 15-hydroxyprostaglandin dehydrogenase (15-PGDH) and phosphomolybdic acid (PMA) was developed. In this assay system, $PGH_2$ was converted to $PGE_2$ by mPGES-1, and then $PGE_2$ was further transformed to the 15-keto-$PGE_2$ by 15-PGDH accompanying the production of NADH, which was easily detected by fluorescence spectrometry in a multi-well plate format. During the reaction, spontaneous oxidation of $PGH_2$ was prevented by PMA. Using this novel assay, the $K_m$ value of mPGES-1 for $PGH_2$ and the $IC_{50}$ value of the previously characterized inhibitor, MK-886, were determined to be 0.150 mM and $2.8\;{\mu}M$, respectively, which were consistent with the previously reported values. In addition, low backgrounds were observed in the multi-wall plate screening of chemical compounds.

Ginsenosides Inhibit Endothelium - dependent Contraction in the Spontaneously Hypertensive Rat Aorta isn vitro (선천성 고혈압 랫드에서 ginsenosides에 의한 내피의존성수축의 억제작용)

  • 김낙두;최원선
    • Journal of Ginseng Research
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    • v.21 no.2
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    • pp.125-132
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    • 1997
  • Our previous study showed that in vivo treatment of spontaneously hypertensive rats (SHR) with protopanaxatriol ginsenosides (PPT) reduces the blood pressure and inhibits the con- tractions induced by endothelium-derived contracting factor (prostaglandin endoperoxide ($PGH_2$) and superoxide anion) in aorta isolated from SHR. The aim of the present study is to examine whether PPT improves endothelial functions in the isolated thoracic aorta of SHR in vitro. Treatments of aortic rings with PPT, purified ginsenoside $Rg_1$ ($Rg_1$) or indomethacin normalized endotheliuln-dependent relaxation to acetylcholine, but not with protopanaxadiol ginsenosides (PPD) and purified ginsenoside Rb1 (Rb1). The effects of PPT were dose-dependent. PGH,- and oxygen free radical-inducted contractions in rat aorta without endothelium were inhibited by PPT or $Rg_1$, but not by PPD or $Rb_1$. Contractions induced by PGF2$\alpha$, U-46619, a stable thromboxane A2 agonist or KCI (60 mM) were not inhibited by PPT, $Rg_1$ or $Rb_1$. These findings demonstrate that PPT but not PPD scavenges the oxygen-derived free radicals and/or antagonize the effects of $PGH_2$ in the vascular smooth muscle and may explain the hypotensive effect of ginseng in the SHR.

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Effect of Protopanaxatriol Ginsenosides on the Blood Pressure and Endothelial Dysfunction In the Aorta of Spontaneously Hypertensive Rats (선천성 고혈압 렛드에서 혈압 및 내피의 기능장해에 대한 protopanaxatriol계 배당체의 효과)

  • 김낙두;김순회
    • Journal of Ginseng Research
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    • v.21 no.2
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    • pp.119-124
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    • 1997
  • Chronic hypertension is associated with impaired endothelial function such as reduced synthesis/release of endothelium-derived relaxing factor(EDRF, nitric oxide) and increased synthesis/release of endothelium-derived contracting factor(EDCF) including prostaglandin endoperoxide($PGH_2$) , superoxide anion both in animals and in humans. We have previously shown that ginsenosides lower the blood pressure and enhance the release of nitric oxide(NO) from endothelial cells in the rat aorta of the normotensive rats. The aim of the present study is to examine whether in vivo treatment of spontaneously hypertensive rats(SHRs) with protopanaxatriol ginsenosides(PPT) reduces the blood pressure and improves endothelial function in the isolated thoracic aorta of SHR. In addition, the contractile response to $PGH_2$ and superoxide anion in the aorta treated with PPT was assessed. SHRs at the age of 16 weeks were savaged with PPT(30 mg/kg/ day) for 2 weeks and systolic blood pressure was measured by the tail-cuff method. Whereas blood pressure was significantly increased in SHRs by 5.4 mmHg during this period of treatment, treatment of SHRs with PPT blocked the elevation of blood pressure. Endothelium-dependent relaxation to acetylcholine was significantly increased in the PPT-treated animals. $PGH_2$- and oxygen-derived free radical-induced contractions were significantly suppressed in aortic rings without endothelium from PPT-treated SHR. These findings indicate that PPT reduces the blood pressure of SHR, which may be associated with either increase of NO release or by antagonizing superoxide anion and PGH2 in the aortic smooth muscle.

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Process Simulation for the Production of Porcine Growth Hormone Using CAD Program (CAD 프로그램을 이용한 돼지 성장호르몬 생산공정모사)

  • 김성구;공인수
    • KSBB Journal
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    • v.10 no.1
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    • pp.97-104
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    • 1995
  • A computer simulation of biochemical process was carried out using Macintosh-based BioDesignerTM developed at Bioprocess and Engineering Center(BPEC) of MIT. Based on the assumptions and flask culture experiments, a porcine growth hormone (PGH, Porcine Somatotropin) production process was simulated by a two-stage continuous culture. The economical and sensitivity analyses were evaluated for the scale-up production of PGH. A high return on investment (ROI, 104%/year) suggested that the process be profitable. However, sensitivity analysis indicated that ROI was dependent on the yield of PGH, selling price, dose and the achievement of projected market penetration.

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Expression and purification of human mPGES-1 in E. coli and identification of inhibitory compounds from a drug-library

  • Kim, Woo-Il;Choi, Kyung-A;Do , Hyun-Soo;Yu, Yeon-Gyu
    • BMB Reports
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    • v.41 no.11
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    • pp.808-813
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    • 2008
  • Human microsomal prostaglandin E synthase-1 (mPGES-1) is a membrane associated protein that catalyzes the conversion of prostaglandin $H_2$ ($PGH_2$) into prostaglandin $E_2$ ($PGE_2$). In this study, the expression of human mPGES-1 in E. coli was significantly enhanced by modifying the utility of specific codons and the recombinant mPGES-1 was efficiently purified to homogeneity. The $K_m$ and $V_{max}$ of the purified enzyme were determined and the trimeric state characterized by chemical cross-linking with glutaraldehyde. The purified mPGES-1 was used for the screening of a chemical library of bioactive or drug compounds to identify novel inhibitors, and oxacillin and dyphylline were identified as moderately inhibiting mPGES-1 with $I_{C50}$ values of 100 and 200 ${\mu}M$, respectively. As these compounds competitively inhibited the catalysis of $PGH_2$, their binding sites appeared to be located near the $PGH_2$ binding pocket.

Refined Exposure Assessment for Three Active Ingredients of Humidifier Disinfectants

  • Lee, Jong-Hyeon;Kang, Hyun-Joong;Seol, Hwi-Soo;Kim, Chan-Kook;Yoon, Seung-Ki;Gwack, Jin;Kim, Yong-Hwa;Kwon, Jung-Hwan
    • Environmental Engineering Research
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    • v.18 no.4
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    • pp.253-257
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    • 2013
  • Exposure assessment for three major active ingredients used for humidifier disinfectants, polyhexamethylene guanidine (PHMG), oligo(2-(2-ethoxy)ethoxyethyl guanidinium chloride (PGH), and 5-chloro-2-methylisothiazol-3(2H)-one/2-methylisothiazol-3(2H)-one (CMIT/MIT) mixture, was conducted in a bedroom using an air sampler for a refined risk assessment. The experimental site was selected to reflect consumer exposure conditions. Aerosols formed by a humidifier were sampled during 8 hr at 7.5 L/min. Absorbed PHMG and PGH by the sampler were quantified using a spectrophotometric method, and high performance liquid chromatography-ultraviolet detection was used for CMIT/MIT. Three exposure scenarios were assumed for adding humidifier disinfectants to the humidifier water at 1, 2, and 10 times the volume recommended by the product suppliers, and the humidifier was on at its maximum rate of producing aerosols in order to consider reasonable worst-cases. The sampled mass of PHMG and PGH ranged 200 to $2,800{\mu}g$ and 140 to $1,900{\mu}g$, respectively, under different exposure conditions, whereas the absorbed mass of CMIT/MIT was barely detected at the detection limit of 0.11/0.29 mg/L, only at 10 times the recommended level. The resulting risk quotients for PHMG and PGH ranged 1,400 to 20,000 and 1,000 to 13,000, indicating that health risks could be significant. For CMIT/MIT mixture, risk quotients were much smaller than estimated by assuming that they are conservative in the indoor environment, probably due to oxidative reactions. The refined exposure assessment presented here may provide a useful tool for assessing risks posed by active ingredients in spray-type biocidal products.

OLED 소자의 효율 개선을 위한 소재 및 구조의 변화에 따른 특성 평가

  • Bae, Il-Ji;Hong, Yeong-Gyu;Yun, Dang-Mo;Sin, Jin-Guk
    • Proceedings of the Korean Vacuum Society Conference
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    • 2014.02a
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    • pp.227-227
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    • 2014
  • OLED 소자에 사용되는 유기물들은 대부분 전자에 비해 정공의 이동속도가 매우 빨라 소자 효율의 손실이 일어난다. 본 연구에서는 이러한 전하 이동도의 불균형에 의한 OLED 소자 성능의 감소를 개선하기 위해 HBL (hole blocking layer) 물질로 BCP (HOMO : 6.5 eV, LUMO : 2.83 eV)를 도입하였다. 그러나 BCP의 LUMO 값이 약 3 eV를 가지기 때문에 전자의 이동에 영향을 미치는 것으로 예상되어 더 높은 효율을 가지는 소자를 제작하기 위해 host 물질을 상용물질(PGH02)로 교체하였다. PGH02의 HOMO 값은 약 5.86 eV로 소자에 사용된 HTL (hole transport layer)의 HOMO 값(5.54 eV)에 비해 높은 값을 가지기 때문에 HBL의 역할 역시 가능하여 소자의 성능이 상당히 개선되는 것을 확인할 수 있었다. 또한 전하 이동도의 균형을 맞추기 위해 ETL 물질로는 기존에 많이 사용되고 있는 Alq3 (${\mu}{\sim}10-5cm2/Vs$)에 비해 이동도가 10배 이상 빠른 Bebq2 (${\mu}{\sim}10-4cm2/Vs$)를 사용하였다. HTL (hole transport layer) 물질로는 상용물질(LHT 259)를 사용하였고, LHT 259의 전하 이동도는 FET (field effect transistor)를 제작하여 측정하였다. 이를 기반으로 하여 ETL과 HTL의 두께를 조절하여 전하 이동도가 균형을 이루는 OLED 소자를 제작하기 위해 실험을 진행하였다.

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A Study on the Relative Motivation of Shannon's Information Theory (샤논 정보이론의 상관성 동기에 관한 연구)

  • Lee, Moon-Ho;Kim, Jeong-Su
    • The Journal of the Institute of Internet, Broadcasting and Communication
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    • v.21 no.3
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    • pp.51-57
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    • 2021
  • In this paper, the relevance between Einstein's special theory of relativity (1905) and Bernoulli's fluid mechanics (1738), which motivates Shannon's theorem (1948), was derived from the AB=A/A=I dimension, and the Shannon's theorem channel code was simulated. When Bernoulli's fluid mechanics ΔP=pgh was applied to the Hallasan volcano Magma eruption, the dimensions and heights matched the measured values. The relationship between Einstein's special theory of relativity, Shannon's information theory, and the stack effect theory of fluid mechanics was analyzed, and the relationship between volcanic eruptions was mathematically proven. Einstein's and Bernoulli's conservation of energy and conservation of mass were the same in terms of bandwidth and power efficiency in Shannon's theorem.