• Biomolecules & Therapeutics
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• v.9 no.2
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• pp.79-87
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• 2001

Heat shock proteins serve as chaperone by preventing the aggregation of denatured proteins and promote survival of pathogens in harsh environments. In this study, heat shock gene encoding a 84-kDa (p84) protein, which is one of the three major heat shock proteins in S. pneumoniae, was cloned and characterized. PCR with a forward primer derived from N-terminal amino acid sequence of the p84 and a reverse primer derived from the conserved second ATP-binding region of Clp family was used for amplification of the gene encoding the p84 and subsequently the PCR product was used for sequence determination. Sequence analysis of the p84 gene demonstrated that it is a member of ClpL. The deduced amino acid sequence of pneumococcal ClpL shows homology with other members of the Clp family, and particularly, even in variable leader region, with bovine Clp-like protein and L. lactis ClpL. S. pneumoniae clpL is the smallest clop member (701 amono acids) containing the two conserved ATP-binding regions, and hydrophilic N-terminal variable region of pneu-mococcal Clp ATPase is much shorter than any known Clp ATPases. Histidine tagged ClpL was overexpressed and purified from E. coli. Immunoblot analysis employing antisera raised against pneumococcus p84 demonstrated no cross-reactivity with Clp analog in Eschericha coli, Staphylococcus aureus and human HeLa cells. Preimmunization of mice with ClpL extended mice life partially but did not protect them from death.

• Proceedings of the PSK Conference
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• 2002.10a
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• pp.336.1-336.1
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• 2002

HSP100/Clp family functions as molecular chaperone and ATP dependent protease. The Streptococcus pneumoniae ClpL. a homologue of bacterial ClpB and yeast cytosolic HSP 104. is one of major heat shock proteins but its biochemical properties are unknown. In this study. ClpL in Streptococcus pneumoniaewas characterized using histidine tagged recombinant ClpL. When ATP hydrolysis activity was compared in the presence or absence of a variety of nucleotides or divalent ions. either ATP or Mn$2^＋$ ion was found to increase significantly the rate of ATP hydrolysis. Furthermore. glutaraldehyde cross-linking and subsequent native-PAGE analfysis showed that ClpL forms dimer. but in the presence of 4 mM concentration of $Mn^{2＋}$ion as a cofactor for ATP hydrolysis and oligormerization in vitro.

• Journal of the East Asian Society of Dietary Life
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• v.21 no.4
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• pp.535-544
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• 2011

The purpose of this study was to investigate the quality characteristics and antioxidative effect of cookies prepared with Curcuma longa L. powder (0, 1, 2, 3, 4%) substituted for flour. The pH of cookie dough decreased significantly in response to the addition of all levels of Curcuma longa L. powder (CLP). However, there were no significant differences in dough values among the test groups. Furthermore, when the spread factor values were compared among the groups, they were found to be inversely proportional to the CLP concentration. In addition, Hunter's color L value significantly decreased with increasing CLP content, whereas the a and b values increased. According to textural measurements, substitution of 1~4% for CLP resulted in increased hardness, cohesiveness, springiness, and brittleness when the samples were compared to the control. The results of sensory evaluation showed that the cookies containing 2% CLP had the highest scores. The acid value, peroxide value, and thiobarbituric acid value were all lower in cookies with 2% and 3% CLP than in cookies made with 4% CLP and control cookies.

• Journal of Korean Society of Occupational and Environmental Hygiene
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• v.25 no.2
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• pp.115-125
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• 2015

Objectives: The major objectives of this study are to review the EU CLP Regulations to propose ways of improving the reliability of MSDS and labels. Methods: To review the EU CLP Regulations, we used EU documents including directives and regulations on chemical management. In addition, we used EU governmental agency reports to illuminate the history and background of the CLP. We found the EU CLP's instruments for reliable hazard communication and evaluated the operations of the instruments. Results: EU CLP Regulations have four instruments for the EU CLP Regulations to make hazard communication reliable. These instruments are GHS, the harmonized CMR and respiratory sensitive substances classification list, C&L inventory and restriction of trade secrets. These are highly useful for achieving the objectives of REACH and CLP(no data-no market and changing the burden of proof). Conclusions: Changing the burden of proof is a key principle for achieving a society safe from hazardous chemicals. Chemical manufacturers and importers alone should bear the responsibility for reliable MSDS. We recommend benchmarking the EU CLP Regulations in order to change efficiently the burden of proof. Trade secrets should be limited to low-hazard substances and be approved by the government before the chemical product is on the market. Like the C&L inventory, chemical product information including substances identification and hazard properties should be notified, aggregated and be opened to public on the Internet. Finally, we recommend a MSDS registration system once again.

• Korean journal of food and cookery science
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• v.25 no.2
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• pp.199-205
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• 2009

Wet noodles with different percentages of Curcuma Ionga L. powder(CLP) as an additive were generated and their cooking characteristics were evaluated. Wheat flour with 8% CLP had the highest water binding capacity and breakdown, while setback was reversed. Cooked noodle characteristics, weight, volume and tensile strength decreased as CLP content of wheat flour increased, but turbidity of the soup was reversed. L value of wet noodles was higher than cooked noodles, while b value, and texture of wet noodles were lower than cooked noodles. Hunter color value and texture measurements demonstrated decreasing L value, springiness and cohesiveness but increasing b value, hardness, gumminess and chewiness with increasing CLP content. In sensory evaluations, noodles made with 4% CLP were most highly preferred, while noodles made with 2% and 8% CLP were less preferred than CLP-free prepared noodles.

• Proceedings of the Korean Society of Applied Pharmacology
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• 2003.11a
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• pp.77-77
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• 2003

Sepsis remains common surgical problems with high morbidity and mortality despite improvement in the management for septic patient. Although hepatocellular dysfunction occurs during sepsis, the mechanism responsible for this remains unclear. In sepsis, a state of severe oxidative stress is encountered, with host endogenous antioxidant defenses overcome. Therefore, the aim of this study was to determine the effect of a-tocopherol (AT) vasoregulatory gene expression during polymicrobial sepsis. Rats were subjected to polymicrobial sepsis by cecal ligation and puncture (CLP). AT (15 mg/kg) was intraperitonealy injected for 3 days prior to CLP. Blood samples were taken 24 h after CLP for measurement of the extent of hepatocellular damage. Liver samples were taken for RT-PCR analysis of mRNA for genes of interest: endothelin-l (ET-l), its receptors $ET_{A}$ and $ET_{B}$, nitric oxide synthases (iNOS and eNOS), cyclooxygenase-2 (COX -2), heme oxygenase-l (HO-l), and tumor necrosis factor-$\alpha$ (TNF-$\alpha$). The activities of serum alanine aminotransferase and lipid peroxidation level were significantly increased; an increase which was prevented by AT pretreatment. CLP significantly increased the mRNA levels of ET-1 and $ET_{B}$; an increase that was prevented by AT pretreatment. However, the level of $ET_{A}$ mRNA significantly decreased after CLP; a decrease that was not prevented by AT pretreatment. There were significant increases in the mRNA expression of iNOS, HO-l and COX -2 in CLP groups. This increase was prevented by AT pretreatment. The expression of eNOS and TNF-$\alpha$ mRNA significantly increased in CLP, which was not prevented by AT pretreatment. Our findings suggest that there was an imbalanced vasoregulatory gene expression in sepsis, and AT ameliorates this change through its free radical scavenging activity.

• Journal of Plant Biotechnology
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• v.30 no.4
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• pp.315-321
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• 2003

Freezing and drought tolerances in plants are very important for survival in the desert. In an effort to reduce desertifcation in Gobi, a molecular breeding of Artemisia adamsii using the CLP (chitinase like protein, antifreeze protein) and Dhn5 (dehydrin5) genes from barley is performed by introducing them into Artemisia adamsii via Agrobacteria. We had found an optimal combinatorial concentration of hormones at 0.05mg/L of NAA and 0.5mg/L of BA for growth of callus in Artemisia adamsii. In addition, the higher rate of callus induction using hypocotyl as explant was observed comparing to explants of stem and leaf. There were some variations in the level of the proteins expressed among the transgenic lines such that the lines of CLP(CS1-5, 1-7,4-4) and Dhn5(DS2-2, 2-3) lines produce the protein to higher levels. The transgenic lines showing a higher level of Dhn5 exhibited better growth than nontransgenic callus in presence of 10 and 20％ PEG. In case of the CLP tansgenic lines, both CS1-5 and CS1-7 showed a higher level of freezing tolerance determined by ion leakage test.

• Proceedings of the Microbiological Society of Korea Conference
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• 2005.05a
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• pp.49-52
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• 2005

• Journal of Trauma and Injury
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• v.21 no.1
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• pp.59-65
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• 2008

Purpose: The aim of this study was to evaluate the effect of overexpression of heat shock protein 70 (HSP70) on the expression of inducible nitric oxide synthase and on the concentration of nitric oxide and to determine the mechanism for the relationship between HSP70 and inducible nitric oxide synthase (iNOS) in sepsis. Methods: Experiments were performed on male Sprague-Dawley rats, and sepsis was induced by using cecal ligation and puncture (CLP). Glutamine (GLN) or saline was administered 1 h after initiation of sepsis. We acquired serum and lung tissues from the rats 12 h or 24 h after initiation of sepsis. We analyzed the concentration of nitric oxide, the expression of HSP70 in the lung, and the gene expression of iNOS in the lung. Results: In CLP+GLN, glutamine given after initiation of sepsis enhanced the expression of HSP70 in the lung at 12 h (CLP+GLN vs. CLP:: $47.19{\pm}10.04$ vs. $33.22{\pm}8.28$, p = 0.025) and 24 h (CLP+GLN vs. CLP: $47.06{\pm}10.60$ vs. $31.90{\pm}4.83$, p = 0.004). In CLP+GLN, glutamine attenuated the expression of iNOS mRNA in the lung at 12 h (CLP+GLN vs. CLP: $4167.17{\pm}951.59$ vs. $5513.73{\pm}1051.60$, p = 0.025) and 24 h (CLP+GLN vs. CLP: $9,437.65{\pm}2,521.07$ vs. $18,740.27{\pm}8,241.20$, p = 0.016) and reduced the concentration of nitric oxide in serum at 12 h (CLP+GLN vs. CLP: $0.86{\pm}0.48$ vs. $3.82{\pm}2.53{\mu}mol/L$, p = 0.016) and 24 h (CLP+GLN vs. CLP: $0.39{\pm}0.25$ vs. $1.85{\pm}1.70{\mu}mol/L$, p = 0.025). Conclusion: The overexpression of HSP70 induced by the administration of glutamine in sepsis attenuated the gene expression of iNOS and reduced the concentration of nitric oxide.

• Journal of Food Hygiene and Safety
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• v.28 no.4
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• pp.342-348
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• 2013

The following study was presented to investigate the anti-inflammatory effect of peel extracts (PE) from three citrus fruits: Citrus unshiu, Citrus limonia Osbeck and Citrus hallabong. According to this study, cytotoxicity, NO-production and protein levels of iNOS (inducible nitric oxide synthase) in macrophage cell were analyzed, which had been incubated in murine macrophage cell line RAW 264.7 cell of PE from those three citrus fruits. According to Citrus unshiu peel extracts (CUP), Citrus limonia Osbeck peel extracts (CHP) and Citrus hallabong peel extracts (CLP) treatment, the result showed that there was no cell growth inhibited below 2 mg/mL. Comparing the NO-production of the cell with LPS (100 ng/mL) and the treatment without LPS, significant increase of NO-production was detected. However NO-production also showed decrease trend, as the concentration increased. For each treatment, at the concentration of 1 mg/mL, NO-ihibitory activity showed significant result with following order: CUP > CHP > CLP. According to the result from Western blot, the inhibitory activities of iNOS protein from CUP and CHP showed fairly similar performances. Also inhibitory activity of COX-2 showed the following order: CUP > CHP> CLP. There was no doubt that all the treatments of CUP, CHP and CLP have anti-inflammatory effect and also that the inhibitory activity of the CUP treatment was the strongest among those three.