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Human CYP1A2 Promoter Fused-Luciferase Gene Constructs Hardly Respond to Polycyclic Hydrocarbons in Transient Transfection Study in HepG2 Cells

  • Chung, Injae
    • Toxicological Research
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    • v.16 no.2
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    • pp.95-100
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    • 2000
  • In previous study, both constitutive expression and 3-methylcholanthrene (3MC)-mediated elevation of CYP1A2 mRNA were demonstrated in human hepatoma HepG2 cells by reverse transcription-polymerase chain reaction (RT-PCR), suggesting that HepG2 cells would be appropriate for the study of human CYP1A2 regulation(Chung and Bresnick, 1994). Further studies were conducted to determine the basis of this induction phenomenon that is observed in HepG2 cells. Since CYP1A1 gene, another polycyclic hydrocarbon(PH)-inducible gene, is regulated by PHs through their interactions via receptors with cis-elements, the 5'-flanking region of human CYP 1A2 gene was analyzed to search such responsive elements. The promoter activity of various lengths of CYP1A2 gene sequence (-3203/+58bp) was measured in transiently-transfected HepG2 cells by fusion constructs containing the CAT, hGH or luciferase genes as a reporter. This region of the CYP1A2 gene, although containing a XRE, was only weakly responsive (less than 2 fold induction) to 10 nM of TCDD or 1 $\mu$M 3 MC treatment. This small enhancement of promoter activity is inconsistent with the previous observation, i.e., 12 to 14 fold-enhanced CYP1A2 mRNA from 1 $\mu$M 3 MC treated HepG2 cells, suggesting that additional mechanisms would exist for PH-mediated induction of CYP1A2 in these cells.

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EFFECT OF CIS-ELEMENT ON THE REGULATION Of TROUT LIVER CYTOCHROME P450IAl GENE EXPRESSION

  • Hwang, Jung E.;Sheen, Yhun Y.
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.188-188
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    • 1996
  • In order to gain insight into the mechanism of the regulation of cytochrome P450IAl by arylhydrocarbon, the 5'-flanking region of a trout CYP450IAl 5'flanking DNA was cloned into pCAT-basic vector and it was transfected into Hepa-1 cells. 3MC treatment to hepa Ⅰ cells transfected with fish CYP450IAl-CAT construct results in mRNA increased by 2.81 fold when it was compared with that of control This increase of mRNA was decreased by concomitantly treated flavonoids such as morin. The levels of CAT mRNA that was treated with morin was 29.2-58.0% of 3MC stimulated CAT mRNA. Further investigation to find out if there are DRE, XRE or negative regulatory cis element in CYP450IA1 gene was undertaken. Results of the deletion study of 5'flanking DNA of trout P450IA indicate the existance of the negative(-1600 ~ -1300). CAT mRNA was about two-fold higher in deleted trout CYP450IAl-CAT construct transfected cells compared to the wi Id type trout CYP450IAl-CAT construct transfected cells. And The stimulatory effect of 3MC was no longer observed in col Is containing deleted CAT construct. [Supported by grants from the Korean Ministry of Education]

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Comparison of Diesel Exhaust Particle Concentration between Large Above-Underground Parking Lots (수도권 일부 대형상가 지상주차장 및 지하주차장의 공기중 디젤엔진배출 입자상물질의 공기중 농도 비교)

  • Kim, Boowook;Song, Dong-Woo
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.23 no.4
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    • pp.323-332
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    • 2013
  • Objectives: This study was conducted in order to investigate the diesel exhaust particle(DEP) concentrations in the thirteen parking lots of large shopping complex. Methods: The real-time black carbon(BC) concentration was determined using an Aethalometer, and elemental/organic carbon concentration was determined according to the method of the National Institute for Occupational Safety and Health(NIOSH) 5040. The particle number concentration(NC), lung deposited surface area concentration(LDSA) and geometric mean diameter(GMD) were determined using a DiSCmini aerosol monitor. Results: The average concentration of BC, EC, OC, NC, LDSA and GMD were $19.1{\mu}g/m^3$, $12.6{\mu}g/m^3$, $51.5{\mu}g/m^3$, $94,000particles/cm^{-3}$, $298{\mu}m^2/cm^{-3}$ and 57 nm in all parking lots, respectively, approximately 3-fold higher than those found in the urban outdoor. The average concentration of BC were $21.3{\mu}g/m^3$ in underground parking lots, 3-fold higher than above parking lots. Conclusions: Therefore, the parking lots at the large shopping complex can be considered a potentially dangerous environment with a high concentration of DEP nanoparticles.

STUDY CYTOCHROME P450IA1 GENE EXPRESSION BY RTPCR.

  • Lee, Soo-Young;Yhun Y. Sheen
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1995.04a
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    • pp.128-128
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    • 1995
  • To investigate the mechanism of the regulation of cytochrome P450IA1 gene expression, ethoxyresorufin deethylase(EROD) and benzo(a)pyrene hydroxylase in B6 mouse liver, in isolated perfused rat liver system. and in B6 mouse hepatocyte Hepa-I cells were examined. In C57BL/6N mouse, 3-methylcholan- throne( 3MC ) treatment have resulted in the stimulation of EROD activity based on fluorometry by 2.79 fold comparirng with that of control. Measurement of mRNA of cytochrome P450 was carried out by either nothern blot or dot blot analysis. Findings are similar to that of studies with enzymes. Furhtermore, when RTPCR method was applied to detect mRNA in Hepa I cell and liver tissues the results were more clear. Cytochrome P450IA1 upstream DNA containing CAT construct was transfected into Hepa-1 cells. After transfection of CAT construct, 3MC and flavonoids, such as, chrysin, hesperetin, kaempferol, morin, myricetin and aminoyrine were treated. 48 Hours after treatments, cells were harvested and assayed for CAT mRNA by RTPCR. 3MC treatment to hepa I cells transfected with trout P450IA1-CAT construct increased CAT mRNA by 2.81 fold when it was compared with that of control. This increase CAT mRNA was decreased by concomitantly treated flavonoids and aminopyrine. The level of CAT protein was 29.2-58.0% of 3MC stimulated CAT protein. Results of this study suggested that RTPCR seems to be a very good method to study regulation of gene expression in liver tissue or Hepa cells.

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Antihypertensive Effects of Enantiomers of Amlodipine Camsylate, a Novel Salt of Amlodipine

  • Oh, Kwang-Seok;Kim, Maeng-Sup;Lee, Byung-Ho
    • Biomolecules & Therapeutics
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    • v.15 no.1
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    • pp.40-45
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    • 2007
  • The vascular relaxant effects on isolated rat aorta of amlodipine camsylates (S-, R-enantiomer, and R/S-racemate), were evaluated and compared with that of S-amlodipine besylate. Furthermore, antihypertensive effects were measured in spontaneously hypertensive rat (SHR). The S-amlodipine camsylate concentration-dependently inhibited $Ca^{2+}$-induced contraction of rat aorta with a very slow onset of action (reached its maximum at 3.5h; $ED_{50}:\;1.50\;{\pm}\;0.24$ nM), having a potency 2-fold higher than those of R/S-amlodipine camsylate $(ED_{50}:\;3.36\;{\pm}\;0.91\;nM)$ and similar to those of S-amlodipine besylate $(ED_{50}:\;1.44\;{\pm}\;0.14\;nM)$, whereas the R-amlodipine camsylate has 590-fold lower vasorelaxant activity $(ED_{50}:\;886.4\;{\pm}\;49.7\;nM)$. In SHR, orally administered S-amlodipine camsylate produced a dose-dependent and long-lasting (>>10 h) antihypertensive effect $(ED_{20}:\;0.89\;mg/kg)$, with a potency 2-fold higher than those of R/S-amlodipine camsylate $(ED_{20}:\;1.82\;mg/kg)$ and similar to those of S-amlodipine besylate $(ED_{20}:\;0.71\;mg/kg)$. In contrast, the R-amlodipine camsylate has no effect even-though administrated high concentration 10 mg/kg. These results suggest that S-amlodipine camsylate has the potency and long-lasting antihypertensive activity as single enantiomer drug, and its antihypertensive effect is not significantly different to that of S-amlodipine besylate.

Clinical implications of the newly defined concept of ventilator-associated events in trauma patients

  • Lee, Tae Yeon;Oh, Jeong Woo;Lee, Min Koo;Kim, Joong Suck;Sohn, Jeong Eun;Wi, Jeong Hwan
    • Journal of Trauma and Injury
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    • v.35 no.2
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    • pp.76-83
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    • 2022
  • Purpose: Ventilator-associated pneumonia is the most common nosocomial infection in patients with mechanical ventilation. In 2013, the new concept of ventilator-associated events (VAEs) replaced the traditional concept of ventilator-associated pneumonia. We analyzed risk factors for VAE occurrence and in-hospital mortality in trauma patients who received mechanical ventilatory support. Methods: In this retrospective review, the study population comprised patients admitted to the Jeju Regional Trauma Center from January 2020 to January 2021. Data on demographics, injury characteristics, and clinical findings were collected from medical records. The subjects were categorized into VAE and no-VAE groups according to the Centers for Disease Control and Prevention/National Healthcare Safety Network VAE criteria. We identified risk factors for VAE occurrence and in-hospital mortality. Results: Among 491 trauma patients admitted to the trauma center, 73 patients who received ventilator care were analyzed. Patients with a chest Abbreviated Injury Scale (AIS) score ≥3 had a 4.7-fold higher VAE rate (odds ratio [OR], 4.73; 95% confidence interval [CI], 1.46-17.9), and those with a glomerular filtration rate (GFR) <75 mL/min/1.73 m2 had 4.1-fold higher odds of VAE occurrence (OR, 4.15; 95% CI, 1.32-14.1) and a nearly 4.2-fold higher risk for in-hospital mortality (OR, 4.19; 95% CI, 1.30-14.3). The median VAE-free duration of patients with chest AIS ≥3 was significantly shorter than that of patients with chest AIS <3 (P=0.013). Conclusions: Trauma patients with chest AIS ≥3 or GFR <75 mL/min/1.73 m2 on admission should be intensively monitored to detect at-risk patients for VAEs and modify the care plan accordingly. VAEs should be closely monitored to identify infections early and to achieve desirable results. We should also actively consider modalities to shorten mechanical ventilation in patients with chest AIS ≥3 to reduce VAE occurrence.

Alterations of Glutathione Peroxidase Patterns by Stressor Treatment in Rice Seedling Roots (스트레스 물질에 의한 벼 glutathione peroxidase 활성패턴 변화)

  • Kim, Yoon-Kyoung;Lee, Mi-Young
    • Applied Biological Chemistry
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    • v.48 no.1
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    • pp.53-59
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    • 2005
  • The effect of various stressors such as reductant ascorbic acid, signalling molecules (salicylic acid and methyl jasmonic acid), heavy metals $(NiCl_2,\;and\;MnSO_4)$ and NaCl on the glutathione peroxidase (GPX) activities and isoenzyme expression patterns were investigated in rice seedling roots. Total GPX activity increased according to the increase of ascorbic acid concentration. Prominent enhancement of GPX1 isozyme due to ascorbic acid contributed to the increase of total GPX activity. GPX showed different reactivity toward salicylic acid and methyl jasmonic acid. GPX activity increased at 0.1 mM salicylic acid, and then decreased thereafter. However, GPX increased gradually in a methyl jasmonic acid concentration-dependent manner, and 3 fold increase of GPX activity was found at 1 mM methyl jasmonic acid. Moreover, GPX1 isozyme increased according to the increase of salicylic acid, while GPX1 isozyme decreased according to the increase of methyl jasmonic acid. When metal ions were treated, GPX activity increased considerably according to the increase of $NiCl_2$ concentration, however, GPX activity increased about 2 fold at 0.5 mM $CuSO_4$ and then decreased. Enhancement of GPX1 isozyme contributed to the increase of total GPX activities in $NiCl_2-treated$ and $MnSO_4-treated$ rice seedlings. Total GPX activity increased 1.7 fold in response to 300 mM NaCl. Especially GPX2 isozyme showed gradual increase according to the increase of NaCl concentration.

Fabrication of TiAl alloy by centrifugal casting and its microstructure (원심주조법에 의한 TiAl 합금의 제조 및 미세구조 분석)

  • Ryu, Jeong Ho;Lee, Ho Jun;Cho, Hyun Su;Paeng, Jong Min;Park, Jong Bum;Lee, Jung-Il
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.27 no.5
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    • pp.229-234
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    • 2017
  • In this study, TiAl alloy was fabricated by a centrifugal casting method for turbo charge of automotive. Optimum conditions for defectless morphology using various ceramic mold were investigated. The crystal structure, microstructure, and chemical composition of the TiAl prepared by centrifugal casting were studied by X-ray diffractometer (XRD), optical microscopy (OM), field emission scanning electron microscopy (FE-SEM) and energy dispersive spectroscopy (EDS), microvickers hardness analyzer (HV). Two kinds of dendrite structures were observed with 4-fold and 6-fold symmetries. The FE-SEM, EDS and HV examinations of the as-cast TiAl showed that the thickness of the oxide layer (${\alpha}$-case) was typically less than $50{\mu}m$.

Dysregulation of Cannabinoid CB1 Receptor Expression in Subcutaneous Adipocytes of Obese Individuals

  • Lee, Yong-Ho;Tharp, William G.;Dixon, Anne E.;Spaulding, Laurie;Trost, Susanne;Nair, Saraswathy;Permana, Paska A.;Pratley, Ridhard E.
    • Animal cells and systems
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    • v.13 no.4
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    • pp.371-379
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    • 2009
  • The endocannabinoid system (ECS) plays a key role in the regulation of appetite, body weight and metabolism. We undertook the present study to further clarify the regulation of the cannabinoid CB1 receptor (CB1, CNR1) in human adipose tissue in obesity. CB1 receptor mRNA expression was ~1.6-fold (p<0.004) and 1.9-fold higher (P<0.05) in subcutaneous adipocytes from obese compared to non-obese subjects in microarray and quantitative real-time PCR studies, respectively. Higher CB1 receptor mRNA expression levels in both adipose tissue (~1.2 fold, P<0.05) and adipocytes (~2 fold, P<0.01) were observed in samples from visceral compared to subcutaneous depots collected from 22 obese individuals. Immunofluorescence confocal microscopy demonstrated the presence of CB1 receptor on adipocytes and also adipose tissue macrophages. These data indicate that adipocyte CB1 receptor is up-regulated in human obesity and visceral adipose tissue and also suggest a potential role for the ECS in modulating immune/inflammation as well as fat metabolism in adipose tissue.

Physicochemical Properties and Antioxidant Activities of Kimchi-added Croquette (김치 첨가 크로켓의 이화학적 품질 특성 및 항산화 활성)

  • Kim, Taejoon;Surh, Jeonghee
    • Journal of the East Asian Society of Dietary Life
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    • v.26 no.6
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    • pp.498-506
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    • 2016
  • A croquette added with heat-treated kimchi at 20% showing higher sensory preferences was analyzed for its physicochemical properties and antioxidant activity using a croquette without kimchi as a control. Compared with the control, kimchi-added croquette had 3.3-fold higher organic acids content (p<0.001), resulting in a significant reduction of pH (p<0.001) and higher metal chelating activity (p<0.001). Upon addition of kimchi, total reducing capacity increased from 109.4 to $139.4{\mu}g/g$ gallic acid equivalents (p<0.01), and DPPH radical scavenging activity also increased 2-fold, which corresponded to 54% of the electron-donating ability of 0.35 mM gallic acid. In addition, contents of free amino acids and ${\gamma}-aminobutyric$ acid (GABA) appreciably increased by 1.6-fold (p<0.01) and 10-fold (p<0.001), respectively. This could be attributed to the ingredients of kimchi and/or enzymatic transformation of precursors by microorganisms during kimchi fermentation. Kimchi-added croquette was determined to be a good source of dietary fiber relative to its calorie content. Texture profile analysis showed no significant differences in hardness, springiness, cohesiveness, gumminess, and chewiness between the two croquettes with or without kimchi. Taken together, this study shows that utilization of heat-treated kimchi as a filling for croquette could be a good strategy to improve both the nutritional quality and antioxidant activity of croquette.