• Korean Journal of Microbiology
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• v.30 no.6
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• pp.528-532
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• 1992

The ATP-dependent protease. Clp P from Esehaichia coli has been increase the stahility with or without detergent as Triton X-100 and NP-40 in the Clp P. The C]p P proteolytic activity was remained to 0.1 M salt by $Na^{-1}$, $K^{+}$, $Li^{+}$ but was inhihited by $SO_4^{2}$. An active ATPase site in Clp A is required for A TP-dependent proteolysis by Clp protease as

• YAKHAK HOEJI
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• v.48 no.1
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• pp.47-54
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• 2004

ClpP is a stress-inducible protein and proteolytic subunit of the ATP-dependent Clp protease in prokaryotes and eukaryotes. Although its physiological roles in bacterial virulence were widely studied in various organsims, including Streptococcus pneumoniae, until now the immunological effect has not been investigated. Here, we have examined the cross reactivity of S. pneumoniae ClpP antibody with other organisms's cell lysate proteins. Although the protein sequence of S. pneumoniae ClpP was highly conserved among various organisms including human, the antibody rasised by S. pneumoniae ClpP was not cross-reacted with other organism's cell lysates, which were Saccharomyces cerevisiae , human lung A549 cell, Bacillus subtilis, Pseuomonas aeruginosa, E. coli, and Salmonella typhi. It was only reacted with S. pneumoniae and Lato-bacillus thermophilus. Thus we examined the immunoprotective effect of ClpP by immunizing mice with the purified ClpP. The mean survival time of mouse was significantly increased with the ClpP immunization. These results suggest that S. pneumoniae ClpP could be used as a vaccine candidate for prevention of S. pneumoniae infection.

• Microbiology and Biotechnology Letters
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• v.21 no.1
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• pp.30-35
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• 1993

Clp is a relatively abundant ATP-dependent protease found in E. coli. Its specific activity was proportional to the concentration of the limiting amount of Clp A and an excess amount of Clp P, and vice versa. Clp A has an intrinsic ATPase activity that is stimulated by casein, and contains a second site for binding A TP, in addition to the ATPase site. The modification of sulfhydryl groups in Clp A with reagents which have bulky groups such as N-phenylmaleimide led to nullifying both ATPase and protease activity. The same sites were modified by sulfhydryl reagents. It seems that the sulfhydryl groups of Clp A are not directly involved in catalysis. Since non-hydrolyzable analogs of ATP do not activate Clp, ATP hydrolysis may be essential for the proteolytic activity of Clp protease. Clp A and Clp P did not associate in the absence of nucleotide. The results suggest that the activity of the proteolytic component, Clp P, is regulated by the A TP-dependent cycling of Clp A between the activator form and the non-activator form.

• 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.

• Journal of Microbiology and Biotechnology
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• v.33 no.9
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• pp.1130-1140
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• 2023

Among the AAA+ proteases in bacteria, FtsH is a membrane-bound ATP-dependent metalloprotease, which is known to degrade many membrane proteins as well as some cytoplasmic proteins. In the intracellular pathogen Salmonella enterica serovar Typhimurium, FtsH is responsible for the proteolysis of several proteins including MgtC virulence factor and MgtA/MgtB Mg²⁺ transporters, the transcription of which is controlled by the PhoP/PhoQ two-component regulatory system. Given that PhoP response regulator itself is a cytoplasmic protein and also degraded by the cytoplasmic ClpAP protease, it seems unlikely that FtsH affects PhoP protein levels. Here we report an unexpected role of the FtsH protease protecting PhoP proteolysis from cytoplasmic ClpAP protease. In FtsH-depleted condition, PhoP protein levels decrease by ClpAP proteolysis, lowering protein levels of PhoP-controlled genes. This suggests that FtsH is required for normal activation of PhoP transcription factor. FtsH does not degrade PhoP protein but directly binds to PhoP, thus sequestering PhoP from ClpAP-mediated proteolysis. FtsH's protective effect on PhoP can be overcome by providing excess ClpP. Because PhoP is required for Salmonella's survival inside macrophages and mouse virulence, these data implicate that FtsH's sequestration of PhoP from ClpAP-mediated proteolysis is a mechanism ensuring the amount of PhoP protein during Salmonella infection.

• 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.

• Proceedings of the Korean Society of Applied Pharmacology
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• 2001.11a
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• pp.109-109
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• 2001

The present study was done to investigate the relationship between Kupffer cells and alteration of cytochrome P-450 (CYP)-dependent drug metabolizing enzyme activities during polymicrobial sepsis. Male rats were subjected to polymicrobial sepsis by cecal ligation and puncture (CLP) followed by fluid resuscitation. The gadolinium chloride (GdC1$_3$, 10 mg/kg), blocker of Kupffer cells, was pretreated intravenously at 48 h and 24 h prior to the induction of CLP. All assay parameters were determined at 24 h after CLP or sham operation. In CLP-treated rats, the mortality rate of animals increased to 50% and serum alanine (ALT) and aspartate aminotransferase (AST) levels also significantly elevated. However, this increase was not suppressed by GdC1$_3$ pretreatment. Microsomal lipid peroxidation markedly increased after CLP operation. This increase was significantly attenuated by pretreatment. Total cytochrome P-450 content and NADPH-cytochrome P-450 reductase activity were not changed after CLP operation, but GdC1$_3$pretreatment reduced total cytochrome P-450 content, The hepatic microsomal CYP 1A1, 1A2, 2Bl and 2El activities in CLP-induced rats were also not significantly different from sham-operated rats. However, GdC1$_3$pretreatment showed a moderate increase in CYP1A1 and 1A2 activities. Our findings suggest that Kupffer cells may be partly responsible for producing hepatocellular dysfunction during sepsis.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.9
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• pp.1296-1302
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• 2009

Ninety six beak-trimmed 72 week-old Lohmann Brown hens were randomly divided into four equal groups. Each group comprised 4 replicates. Isoenergetic and isonitrogenous experimental diets contained low calcium (3.5%); optimum calcium (4.2%); low Ca (3.5% Ca)+1% Clinoptilolite (CLP); low Ca (3.5% Ca)+2% CLP. Data were collected biweekly and the experiment lasted 6 weeks. Egg production, feed consumption, feed conversion ratio, egg weight, tibia Ca, P, ash and eggshell thickness were not affected by addition of CLP to the diets (p>0.05). There were no significant differences in egg shell strength and ash when data were analyzed individually in measurement periods ($74^{th}$, $76^{th}$ and $78^{th}$ weeks). However, according to pooled data ($74^{th}$-$78^{th}$ weeks), eggshell strength was increased (p<0.05) only by 2% CLP supplementation versus low Ca (3.5%) diet, and shell ash was significantly increased by 2% CLP supplementation compared with the other diets. The damaged egg ratio on 1% and 2% CLP diets was significantly decreased between 76-78 weeks'data when compared with the low Ca diet. However; damaged egg ratio on the 2% CLP diet was significantly decreased when pooled data (74-78) were compared with no CLP diets. The differences in marketable egg ratio paralleled damaged egg ratio. The plasma calcium level at the end of experiment was increased on the 2% CLP diet when compared with the low Ca (3.5%) diet (p<0.05). Furthermore, at the end of the experiment a marked decrease of manure moisture was observed on both CLP diets (p<0.01). In conclusion, Clinoptilolite (2%) supplementation to layer diets tends to improve eggshell quality and manure dry matter (1% and 2% CLP) after six weeks.

• The korean journal of orthodontics
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• v.48 no.4
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• pp.216-223
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• 2018

Objective: This study is performed to investigate the trend of health care (HC) utilization among cleft lip and/or palate (CL/P) during 2007-2016 by using data from the Korean National Health Insurance Service (KNHIS). Methods: The KNHIS data were reorganized to count a specific patient only once for a specific year. Cleft type (cleft lip [CL], cleft palate [CP], and cleft lip and palate [CLP]), sex, and age at HC utilization were investigated. The study period was divided into the first half (2007-2011) and the last half (2012-2016). The utilization number and rate per 1,000 population were calculated for the total population and for new-born patients. Independent t-test and one-way analysis of variance were used for statistical analyses. Results: The total CL/P population (n = 48,707) comprised 19.2% CLP, 35.5% CL, and 45.3% CP (CLP < CL < CP; p < 0.001). Their HC utilization rate increased from 0.066 in 2007 to 0.118 in 2016. The new-born patient population (n = 7,617) comprised 18.6% CLP, 30.4% CL, and 51.0% CP (CLP < CL < CP; p < 0.001). Their HC utilization rate increased from 1.12 in 2007 to 1.74 in 2016. An examination of the utilization number and rate among new-born patients revealed CP exhibited a female-dominant pattern (all p < 0.01), while CL and CLP exhibited a male-dominant pattern (all p < 0.01). However, utilization number showed no difference by sex and cleft type between 2007-2011 and 2012-2016. Conclusions: These results might serve as a guideline for HC utilization among patients with CL/P.

• The Korean Journal of Mycology
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• v.24 no.4 s.79
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• pp.310-321
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• 1996

Five strains AF027, AF069, AF2001, AF2011 and SD02 of Pseudomonas cepacia were isolated from soil, and the antifungal cyclic lipopeptides(CLP) i.e, CLP027A, CLP069A, Cepacidine A, CLP2011A and CLP02A were produced from each strains, respectively. Nitrogen and carbon sources in media were proved to be important factors for the production of CLP and among them, polypeptone-S, glucose and fructose were the most effective. It appeared that compounds CLP027A and CLP069A were identical with Cepacidine A and Xylocandine A, respectively. contain aspartic acid as amino acid component, are differentiated from Xylocandine A containing asparagine. Although molecular weight, amino acid composition and UV spectrum of CLP2011A and CLP02A are same with those of Cepacidine A, it is postulated that these compounds are not identical with Cepacidine A when the antifungal spectra and antifungal activity were compared to those of Cepacidine A.