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Can Urinary Cotinine Predict Nicotine Dependence Level in Smokers?

  • Jung, Hyun-Suk;Kim, Yeol;Son, Jungsik;Jeon, Young-Jee;Seo, Hong-Gwan;Park, So-Hee;Huh, Bong Ryul
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.11
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    • pp.5483-5488
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    • 2012
  • Background: Although nicotine dependence plays a role as a main barrier for smoking cessation, there is still a lack of solid evidence on the validity of biomarkers to determine nicotine dependence in clinical settings. This study aimed to investigate whether urinary cotinine levels could reflect the severity of nicotine dependence in active smokers. Materials and Methods: Data regarding general characteristics and smoking status was collected using a self-administered smoking questionnaire. The Fagerstr$\ddot{o}$m test for nicotine dependence (FTND) was used to determine nicotine dependence of the participants, and a total of 381 participants were classified into 3 groups of nicotine dependence: low (n=205, 53.8%), moderate (n=127, 33.3%), and high dependence groups (n=49, 12.9%). Stepwise multiple linear regression model and receiver operating characteristic (ROC) curves analyses were used to determine the validity of urinary cotinine for high nicotine dependence. Results: In correlation analysis, urinary cotinine levels increased with FTND score (r=0.567, P<0.001). ROC curves analysis showed that urinary cotinine levels predicted the high-dependence group with reasonable accuracy (optimal cut-off value=1,000 ng/mL; AUC=0.82; P<0.001; sensitivity=71.4%; specificity=74.4%). In stepwise multiple regression analysis, the total smoking period (${\beta}$=0.042, P=0.001) and urinary cotinine levels (${\beta}$=0.234, P<0.001) were positively associated with nicotine dependence, whereas an inverse association was observed between highest education levels (>16 years) and nicotine dependence (${\beta}$=-0.573, P=0.034). Conclusions: The results of this study support the validity of using urinary cotinine levels for assessment of nicotine dependence in active smokers.

Screening of Anti-Inflammatory Herbs having the Activation of MAPK Family Proteins (MAPKs의 활성화 기전을 나타내는 항염증 소재의 탐색)

  • Lee, Keyong Ho;Rhee, Ki-Hyeong
    • The Korean Journal of Food And Nutrition
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    • v.28 no.3
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    • pp.343-350
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    • 2015
  • The aim of this study was to evaluate the potential anti-inflammorty properties of herbs via MAPKs such as ERK, p38, JNK. Fifty-one kinds of each herbal medicine, that were extracted with ethanol, were used in the inhibitory assay of cytokine (TNF-${\alpha}$, IL-$1{\beta}$ and IL-6) and NO. of these, 10 species of herbal medicines, Angelica dahurica, Atractylodes lancea, Cnidium officinale, Duchesnea chrysantha, Oldenlandia diffusa, Lonicera japonica, Paeonia lactiflora, Pinus thunbergii, Rehmannia glutinosa and Rubus coreanus, were screened as potential inhibitors (< $300{\mu}g/mL$) of NO, TNF-${\alpha}$, IL-$1{\beta}$ and IL-6. Among the 10 species, Lonicera japonica showed potential anti-inflammatory effects Lonicera japonica extract of $200{\mu}g/mL$ inhibited the phosphorylation of ERK, p38 and JNK. In addition, Lonicera japonica extract at 20 mg/kg increased survival rate from LPS-induced endotoxin shock by 3 fold.

Economic Analsys of Cooling-Heating System Using Ground Source Heat in Multi Family Apartment (공동주택에서 지열 냉난방 시스템 적용시 경제성 분석)

  • Park, Yongboo;Park, Jongbae;Lim, Haesik;Baek, Sungkoon
    • Journal of the Korean GEO-environmental Society
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    • v.8 no.3
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    • pp.11-18
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    • 2007
  • This paper studied the economical efficiency of ground source heat pump system under various conditions in apartments which have important effects on the housing market. And this study analysed the initial cost increase, saved managing cost and recovery time of initial cost. Analysis result showed as time of heating-cooling and water heating increases, the amount of saved managing cost increased much than the initial construction cost, so recovery time shortened. And as the net area of apartment increases, the recovery time increased. The study of the relation between the installation type and recovery time of initial construction cost showed when heat-cooling system adapted ground source heat and water heating system adapted waste heat, the initial construction cost was recovered most quickly. When Ground Source Heating system was used for the heating-cooling and water heating system, ground source heating system was used for the heating-cooling and waste heat used for water heating, and ground source heating system was used for the heating-cooling and LNG used for water heating, the construction cost increased 72,000, 66,900 and 62,300 won each per $m^2$ compared to the current system (package air-conditioner, heating and water heating using LNG).

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Inhibitory effects of polyphenols isolated from Rhus verniciflua on Aldo-keto reductase family 1 B10

  • Song, Dae-Geun;Lee, Joo-Young;Lee, Eun-Ha;Jung, Sang-Hoon;Nho, Chu-Won;Cha, Kwang-Hyun;Koo, Song-Yi;Pan, Cheol-Ho
    • BMB Reports
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    • v.43 no.4
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    • pp.268-272
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    • 2010
  • Aldo-keto reductase family 1 B10 (AKR1B10) is a member of the NADPH-dependent aldo-keto reductase (AKR) superfamily, and has been considered to be a potential cancer therapeutic target. Total extract from the bark of Rhus verniciflua (Toxicodendron vernicifluum (Stokes)) showed AKR1B10 inhibitory activity. To identify the active compounds from R. verniciflua responsible for AKR1B10 inhibition, nine compounds were isolated via bioactivity-guided isolation and tested for their effects against recombinant human AKR1B10 (rhAKR1B10). Results showed that butein, isolated from the ethyl acetate fraction, was most able to inhibit rhAKR1B10. The inhibitory rate of butein against rhAKR1B10 was 42.86% at $1\;{\mu}M$ with an IC50 value of $1.47\;{\mu}M$, and enzyme kinetic analysis revealed its inhibition mode to be uncompetitive.

ATF3 Mediates Anti-Cancer Activity of Trans-10, cis-12-Conjugated Linoleic Acid in Human Colon Cancer Cells

  • Kim, Kui-Jin;Lee, Jihye;Park, Yeonhwa;Lee, Seong-Ho
    • Biomolecules & Therapeutics
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    • v.23 no.2
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    • pp.134-140
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    • 2015
  • Conjugated linoleic acids (CLA) are a family of isomers of linoleic acid. CLA increases growth arrest and apoptosis of human colorectal cancer cells through an isomer-specific manner. ATF3 belongs to the ATF/CREB family of transcription factors and is associated with apoptosis in colorectal cancer. The present study was performed to investigate the molecular mechanism by which t10, c12-CLA stimulates ATF3 expression and apoptosis in human colorectal cancer cells. t10, c12-CLA increased an apoptosis in human colorectal cancer cells in dose dependent manner. t10, c12-CLA induced ATF3 mRNA and luciferase activity of ATF3 promoter in a dose-dependent manner. The responsible region for ATF3 transcriptional activation by t10, c12-CLA is located between -147 and -1850 of ATF3 promoter. mRNA stability of ATF3 was not affected by t10, c12-CLA treatment. t10, c12-CLA increases $GSK3{\beta}$ expression and suppresses IGF-1-stimulated phosphorylation of Akt. The knockdown of ATF3 suppressed expression of $GSK3{\beta}$ and NAG-1 and PARP cleavage. The results suggest that t10, c12-CLA induces apoptosis through ATF3-mediated pathway in human colorectal cancer cells.

DNA Binding Specificity of Proteus mirabilis Transcription Regulator (Proteus mirabilis 전사 조절 단백질의 DNA 결합 특성)

  • Gang, Jong-Back
    • Korean Journal of Microbiology
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    • v.47 no.2
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    • pp.158-162
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    • 2011
  • Amino acid sequence alignment shows that $\underline{P}$roteus $\underline{m}$irabilis $\underline{t}$ranscription $\underline{r}$egulator (PMTR) has cystein sequence homology at metal binding domain to CueR (copper resistance) protein, which conserves two cysteins (Cys 112 and Cys 120 in PMTR). Gel shift assay revealed that PMTR protein bound to promoter region of Escherichia coli copA (copper-translocating P-type ATPase) and Proteus mirabilis atpase (putative copper-translocating P-type ATPase) genes except that of E. coli zntA (zinc-translocating P-type ATPase) gene. DNase I protection experiment indicated that PMTR protein protected the region over -35 box and close to -10 box. DNase I hypersensitive bases were shown at C and A bases of labeled template strand and at G and C bases of labeled non-template strand of DNA. These hypersensitive bases were appeared in other metalloregulatory proteins of MerR family, which suggests protein-induced DNA bending.

Fucoidan Induces Apoptosis in A2058 Cells through ROS-exposed Activation of MAPKs Signaling Pathway

  • Ryu, Yea Seong;Hyun, Jin Won;Chung, Ha Sook
    • Natural Product Sciences
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    • v.26 no.3
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    • pp.191-199
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    • 2020
  • Fucoidan, a natural component of brown seaweed, has various biological activities such as anti-cancer activity, anti-oxidant, and anti-inflammatory against various cancer cells. However, the fucoidan has been implicated in melanoma cells via apoptosis signaling pathway. Therefore, we investigated apoptosis with fucoidan in A2058 human melanoma cells with dose- and time-dependent manners. In our results, A2058 cells viability decreased at relatively short-time and low-concentration through fucoidan. This effects of fucoidan on A2058 cells appeared to be mediated by the induction of apoptosis, as manifested by morphological changes through DNA-binding dye Hoechst 33342 staining. When a dose of 80 ㎍/mL fucoidan was treated, the cells were observed: crescent or ring-like structure, chromatin condensation, and nuclear fragmentation. With the increase at 100 ㎍/mL fucoidan, the cell membrane is intact throughout the total process, including membrane blebbing and loss of membrane integrity as well as increase of sub-G1 DNA. Furthermore, to understand the exact mechanism of fucoidan-treated in A2058 cells, western blotting was performed to detect apoptosis-related protein expression. In this study, Bcl-2 family proteins can be regulated by fucoidan, suggesting that fucoidan-induced apoptosis is modulated by intrinsic pathway. Therefore, expression of Bcl-2 and Bax may result in altered permeability, activating caspase-3 and caspase-9. And the cleaved form of poly ADP-ribose polymerase was detected in fucoidan-treated A2058 cells. These results suggest that A2058 cells are highly sensitive to growth inhibition by fucoidan via apoptosis, as evidenced by activation of extracellular signal-regulated kinases/p38/Bcl-2 family signaling, as well as alteration in caspase-9 and caspase-3.

Characterization of Novel Salt-Tolerant Esterase Isolated from the Marine Bacterium Alteromonas sp. 39-G1

  • Won, Seok-Jae;Jeong, Han Byeol;Kim, Hyung-Kwoun
    • Journal of Microbiology and Biotechnology
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    • v.30 no.2
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    • pp.216-225
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    • 2020
  • An esterase gene, estA1, was cloned from Alteromonas sp. 39-G1 isolated from the Beaufort Sea. The gene is composed of 1,140 nucleotides and codes for a 41,190 Da protein containing 379 amino acids. As a result of a BLAST search, the protein sequence of esterase EstA1 was found to be identical to Alteromonas sp. esterase (GenBank: PHS53692). As far as we know, no research on this enzyme has yet been conducted. Phylogenetic analysis showed that esterase EstA1 was a member of the bacterial lipolytic enzyme family IV (hormone sensitive lipases). Two deletion mutants (Δ20 and Δ54) of the esterase EstA1 were produced in Escherichia coli BL21 (DE3) cells with part of the N-terminal of the protein removed and His-tag attached to the C-terminal. These enzymes exhibited the highest activity toward p-nitrophenyl (pNP) acetate (C2) and had little or no activity towards pNP-esters with acyl chains longer than C6. Their optimum temperature and pH of the catalytic activity were 45℃ and pH 8.0, respectively. As the NaCl concentration increased, their enzyme activities continued to increase and the highest enzyme activities were measured in 5 M NaCl. These enzymes were found to be stable for up to 8 h in the concentration of 3-5 M NaCl. Moreover, they have been found to be stable for various metal ions, detergents and organic solvents. These salt-tolerant and chemical-resistant properties suggest that the enzyme esterase EstA1 is both academically and industrially useful.

Gene Cloning, Expression, and Characterization of a $\beta$-Agarase, AgaB34, from Agarivorans albus YKW-34

  • Fu, Xiao Ting;Pan, Cheol-Ho;Lin, Hong;Kim, Sang-Moo
    • Journal of Microbiology and Biotechnology
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    • v.19 no.3
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    • pp.257-264
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    • 2009
  • A $\beta$-agarase gene, agaB34, was functionally cloned from the genomic DNA of a marine bacterium, Agarivorans albus YKW-34. The open reading frame of agaB34 consisted of 1,362 bp encoding 453 amino acids. The deduced amino acid sequence, consisting of a typical N-terminal signal peptide followed by a catalytic domain of glycoside hydrolase family 16 (GH-16) and a carbohydrate-binding module (CBM), showed 37-86% identity to those of agarases belonging to family GH-16. The recombinant enzyme (rAgaB34) with a molecular mass of 49 kDa was produced extracellularly using Escherichia coli $DH5{\alpha}$ as a host. The purified rAgaB34 was a $\beta$-agarase yielding neoagarotetraose (NA4) as the main product. It acted on neoagarohexaose to produce NA4 and neoagarobiose, but it could not further degrade NA4. The maximal activity of rAgaB34 was observed at $30^{\circ}C$ and pH 7.0. It was stable over pH 5.0-9.0 and at temperatures up to $50^{\circ}C$. Its specific activity and $k_{cat}/K_m$ value for agarose were 242 U/mg and $1.7{\times}10^6/sM$, respectively. The activity of rAgaB34 was not affected by metal ions commonly existing in seawater. It was resistant to chelating reagents (EDTA, EGTA), reducing reagents (DTT, $\beta$-mercaptoethanol), and denaturing reagents (SDS and urea). The E. coli cell harboring the pUC18-derived agarase expression vector was able to efficiently excrete agarase into the culture medium. Hence, this expression system might be used to express secretory proteins.

A Novel Glycosyl Hydrolase Family 16 β-Agarase from the Agar-Utilizing Marine Bacterium Gilvimarinus agarilyticus JEA5: the First Molecular and Biochemical Characterization of Agarase in Genus Gilvimarinus

  • Lee, Youngdeuk;Jo, Eunyoung;Lee, Yeon-Ju;Hettiarachchi, Sachithra Amarin;Park, Gun-Hoo;Lee, Su-Jin;Heo, Soo-Jin;Kang, Do-Hyung;Oh, Chulhong
    • Journal of Microbiology and Biotechnology
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    • v.28 no.5
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    • pp.776-783
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    • 2018
  • The agarase gene gaa16a was identified from a draft genome sequence of Gilvimarinus agarilyticus JEA5, an agar-utilizing marine bacterium. Recently, three agarase-producing bacteria, G. chinensis, G. polysaccharolyticus, and G. agarilyticus, in the genus Gilvimarinus were reported. However, there have been no reports of the molecular characteristics and biochemical properties of these agarases. In this study, we analyzed the molecular characteristics and biochemical properties of agarases in Gilvimarinus. Gaa16A comprised a 1,323-bp open reading frame encoding 441 amino acids. The predicted molecular mass and isoelectric point were 49 kDa and 4.9, respectively. The amino acid sequence of Gaa16A showed features typical of glycosyl hydrolase family 16 (GH16) ${\beta}$-agarases, including a GH16 domain, carbohydrate-binding region (RICIN domain), and signal peptide. Recombinant Gaa16A (excluding the signal peptide and carbohydrate-binding region, rGaa16A) was expressed as a fused protein with maltose-binding protein at its N-terminus in Escherichia coli. rGaa16A had maximum activity at $55^{\circ}C$ and pH 7.0 and 103 U/mg of specific activity in the presence of 2.5 mM $CaCl_2$. The enzyme hydrolyzed agarose to yield neoagarotetraose as the main product. This enzyme may be useful for industrial production of functional neoagaro-oligosaccharides.