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( Nari Lee ),( Hyeongseok Yun ),( Chan Lee ),( Yikjae Lee ),( Euna Kim ),( Sumi Kim ),( Hyoeun Jeon ),( Chiho Yu ),( Jaerang Rho ) 한국미생물생명공학회(구 한국산업미생물학회) 2021 Journal of microbiology and biotechnology Vol.31 No.1
Organophosphorus nerve agents (OPNAs), including both G- and V-type nerve agents such as sarin, soman, tabun and VX, are extremely neurotoxic organophosphorus compounds. Catalytic bioscavengers capable of hydrolyzing OPNAs are under development because of the low protective effects and adverse side effects of chemical antidotes to OPNA poisoning. However, these bioscavengers have certain limitations for practical application, including low catalytic activity and narrow specificity. In this study, we generated a fusion-hybrid form of engineered recombinant human paraoxonase 1 (rePON1) and bacterial organophosphorus hydrolase (OPH), referred to as GV-hybrids, using a flexible linker to develop more promising catalytic bioscavengers against a broad range of OPNAs. These GV-hybrids were able to synergistically hydrolyze both G-type OPNA analogs (paraoxon: 1.7 ~ 193.7-fold, p-nitrophenyl diphenyl phosphate (PNPDPP): 2.3 ~ 33.0-fold and diisopropyl fluorophosphates (DFP): 1.4 ~ 22.8-fold) and V-type OPNA analogs (demeton-S-methyl (DSM): 1.9 ~ 34.6-fold and malathion: 1.1 ~ 4.2-fold above) better than their individual enzyme forms. Among the GV-hybrid clones, the GV7 clone showed remarkable improvements in the catalytic activity toward both G-type OPNA analogs (k<sub>cat</sub>/K<sub>m</sub> (10<sup>6</sup> M<sup>-1</sup> min<sup>-1</sup>): 59.8 ± 0.06 (paraoxon), 5.2 ± 0.02 (PNPDPP) and 47.0 ± 6.0 (DFP)) and V-type OPNA analogs (k<sub>cat</sub>/K<sub>m</sub> (M<sup>-1</sup> min<sup>-1</sup>): 504.3 ± 48.5 (DSM) and 1324.0 ± 47.5 (malathion)). In conclusion, we developed GV-hybrid forms of rePON1 and bacterial OPH mutants as effective and suitable catalytic bioscavengers to hydrolyze a broad range of OPNA analogs.
Chevron 유로 내의 미시적 해석 결과를 통한 대형 판형열교환기 특성에 대한 준미시적 해석
이나리(Nari Lee),이명성(Myungsung Lee),이상혁(Sang Hyuk Lee),허남건(Nahmkeon Hur) 대한설비공학회 2009 대한설비공학회 학술발표대회논문집 Vol.2009 No.-
In the present study, the flow and heat transfer characteristics of a large plate heat exchanger are investigated numerically. The flow passages are very complicated due to the grooved corrugation patterns of the plate surface so that the detailed mesh and the large amount of the computation time have to be required in the numerical simulation for the conjugate heat transfer analysis. In order to accomplish the efficient and fast analysis of the heat transfer characteristics in the plate heat exchanger, a semimicroscopic method using the porous media model has been investigated numerically. The results showed that the characteristics of the heat transfer and pressure drop, which are respectively presented with Colburn j-factor and Fanning f-factor, are in a good agreement between the detailed mesh and the porous media model. The results of the present study could be applicable to the numerical analysis of entire flow passages in the large plate heat exchanger using porous media treatment.
Jae-Geun Lee,Soohyun Lee,Juhee Jeon,Hyun Gi Kong,Hyun-Ju Cho,Jong-Hwan Kim,Seon-Young Kim,Myung Jin Oh,Daum Lee,Nari Seo,Ki Hun Park,Kweon Yu,Hyun Joo An,Choong-Min Ryu,Jeong-Soo Lee 한국당과학회 2022 한국당과학회 학술대회 Vol.2022 No.07
Host tp53 mutations are frequently found during the early stages of colitis-associated colorectal cancer (CAC), but whether such mutations induce gut microbiota dysbiosis and chronic intestinal inflammation that contributes to the development of CAC, remains unknown. We found that zebrafish tp53 mutant larvae exhibited elevated intestinal inflammation, by monitoring the NFκB activity in the mid-distal intestines of zebrafish larvae using an NFκB:EGFP transgenic reporter line in vivo as well as neutrophil infiltration into the intestine. This inflammation was due to dysbiotic gut microbiota with reduced diversity, revealed using both 16S rRNA amplicon sequencing and a germfree larva model. In this dysbiosis, Aeromonas spp. were aberrantly enriched as major pathobionts and exhibited the capacity for aggressive colonization in tp53 mutants. Importantly, the ex-germfree experiments supported the causality of the host tp53 mutation for inducing the inflammation. Transcriptome and high performance liquid chromatography analyses of the host gastrointestinal tracts identified dysregulated sialic acid (SA) metabolism concomitant with increased host Neu5Gc levels as the key determinant of aberrant inflammation, which was reversed by the sialidase inhibitors oseltamivir and Philippin A. These results demonstrate a crucial role for host tp53 in maintaining symbiosis and immune homeostasis via SA metabolism. Disturbed SA metabolism via a tp53 mutation may be exploited by specific elements of the gut microbiome, eliciting both dysbiosis and inflammation. Manipulating sialometabolism may therefore provide an efficacious therapeutic strategy for tp53 mutation-induced dysbiosis, inflammation, and, ultimately, related cancers.
Lee, Sung Ryul,Lee, Seon Joong,Kim, Soon Ha,Ko, Kyung Soo,Rhee, Byoung Doo,Xu, Zhelong,Kim, Nari,Han, Jin Published for the International Federation for Cel 2014 Cell biology international Vol.38 No.6
<P>Although sodium nitroprusside (SNP) is an effective hypotensive drug and is often used in pediatric intensive care units and to treat acute heart failure, clinical application of SNP is limited by its cardiotoxicity. NecroX-5 (NX-5) was recently developed and has the capacity to inhibit necrotic cell death. No current literature addresses whether NX-5 suppresses SNP-induced cell death or its mechanism of action. We have investigated the protective role of NX-5 against SNP-induced cell death in cardiomyocyte-like H9c2 cells. SNP treatment induced severe cell death, possibly through phosphorylation of stress-activated protein kinase/c-Jun NH2-terminal kinase (JNK) and activation of the apoptotic signaling pathway, including downregulation of Bcl-2 and cleavage of caspase-3. However, NX-5 suppresses SNP-induced cell death through inhibition of JNK activation and suppression of both downregulation of Bcl-2 protein expression and caspase-3 cleavage. These findings will provide insights and facilitate development of antidotes to SNP toxicity in cardiac cells.</P>
Lee,Nari Min,Kyung Hee Kim,Hye-Jin 숙명여자대학교 자연과학연구소 1999 자연과학논문집 Vol.- No.10
A 3.0 kb LA PCR product carrying the nahC, nahF and partial nahB gene for upper naphthalene catabolism was generated from the chromosomal DNA of Pseudomonas fluorerscens SME11. This product was inserted into pT7Blue(R) vector, resulting recombinant DNA was named as pNN1 , Restriction endonuclease mapping of 3.0 kb insert of the pNNl was carried out with ApaI, EcoRI, PstI, HindⅢ, SalI, BamHl, KpnI, and SphI. By means of bidirectional subcloning and dideoxynucleotide chain termination, the nucleotide sequences of the pNNF containing nahF and partial nahB genes were determined. A complete ORF (open reading frame) of 1449bp corresponding to 483 amino acids was determined. Deduced amino acid sequences of the nahF gene showed about 96%, 93%, 90%. 85%, and 63% homologies to those of salicylaldehyde dehydrogenase corresponding genes of phaF of P. putida, nahF of P. putida. phaF of P. aeruginosa, nagF of Pseudomonas. sp. U2, and phnF of Burkholderia sp. RP007, respectively. The intact DNA fragment was isolated from the chromosomal DNA by Southern hybridization, and expression of nahF gene was investigated on SDS-PAGE. Salicylaldehyde dehydrogenase를 암호하는 유전자는 Pseudomonas fluorescens SMEll의 염색체 DNA를 주형으로, nahC 유전자 내부 염기서열을 primer로, LA PCR을 실행하여 얻은 3.0 kb DNA로 클로닝되었다. 이 절편을 pT7Blue(R) 벡터에 삽입시켜 재조합 플라스미드를 얻었으며, 이 재조합 플라스미드를 pNNl이라 명명하였다. 조합 DNA를 제한효소로 처리하여 ApaI, EcoRI, PstI HindⅢ, SalI, BamHI, KpnI, SphI 등이 존재함을 확인하였으며, 이것으로 간단한 제한효소지도를 작성하였다. Insert DNA를 서브클로닝하여 3.0 kb DNA 절편의 염기서열을 양방향으로 읽어 결정하였다. 염기서열 분석을 통해 pNNl에는 1449 bp의 완전한 ORF(open reading frame)가 존재하며, 이는 salicylaldehyde dehydrogenase에 해당하는 483개의 아미노산을 암호함을 알 수 있었다. 아울러 SDS-PAGE방법으로 NahF단백질의 발현을 확인하였다. 이미 보고된 P. putides와 Pseudomonas sp. U2의 두가지 salicylaldehyde dehydrogenase의 아미노산서열들과 비교한 결과 각각 93%, 85%의 homology를 보였으며, dehydrogenase 계열의 아미노산 서열과 비교해 본 결과 96%, 90%, 63%의 homology를 보였다. Southern hybridization으로 intact DNA를 다시 클로닝하였으며 SDS- PAGE에서 nahF 유전자의 발현도 검토하였다.
Lee,Nari,Min,Kyung Hee,Kim,Hye-Jin 숙명여자대학교 자연과학연구소 1999 자연과학논문집 Vol.- No.10
Salicylaldehyde dehydrogenase를 암호하는 유전자는 Pseudomonas fluorescens SMEll의 염색체 DNA를 주형으로, nahC 유전자 내부 염기서열을 primer로, LA PCR을 실행하여 얻은 3.0 kb DNA로 클로닝되었다. 이 절편을 pT7Blue(R) 벡터에 삽입시켜 재조합 플라스미드를 얻었으며, 이 재조합 플라스미드를 pNNl이라 명명하였다. 조합 DNA를 제한효소로 처리하여 ApaI, EcoRI, PstI HindⅢ, SalI, BamHI, KpnI, SphI 등이 존재함을 확인하였으며, 이것으로 간단한 제한효소지도를 작성하였다. Insert DNA를 서브클로닝하여 3.0 kb DNA 절편의 염기서열을 양방향으로 읽어 결정하였다. 염기서열 분석을 통해 pNNl에는 1449 bp의 완전한 ORF(open reading frame)가 존재하며, 이는 salicylaldehyde dehydrogenase에 해당하는 483개의 아미노산을 암호함을 알 수 있었다. 아울러 SDS-PAGE방법으로 NahF단백질의 발현을 확인하였다. 이미 보고된 P. putides와 Pseudomonas sp. U2의 두가지 salicylaldehyde dehydrogenase의 아미노산서열들과 비교한 결과 각각 93%, 85%의 homology를 보였으며, dehydrogenase 계열의 아미노산 서열과 비교해 본 결과 96%, 90%, 63%의 homology를 보였다. Southern hybridization으로 intact DNA를 다시 클로닝하였으며 SDS- PAGE에서 nahF 유전자의 발현도 검토하였다. A 3.0 kb LA PCR product carrying the nahC, nahF and partial nahB gene for upper naphthalene catabolism was generated from the chromosomal DNA of Pseudomonas fluorerscens SME11. This product was inserted into pT7Blue(R) vector, resulting recombinant DNA was named as pNN1 , Restriction endonuclease mapping of 3.0 kb insert of the pNNl was carried out with ApaI, EcoRI, PstI, HindⅢ, SalI, BamHl, KpnI, and SphI. By means of bidirectional subcloning and dideoxynucleotide chain termination, the nucleotide sequences of the pNNF containing nahF and partial nahB genes were determined. A complete ORF (open reading frame) of 1449bp corresponding to 483 amino acids was determined. Deduced amino acid sequences of the nahF gene showed about 96%, 93%, 90%. 85%, and 63% homologies to those of salicylaldehyde dehydrogenase corresponding genes of phaF of P. putida, nahF of P. putida. phaF of P. aeruginosa, nagF of Pseudomonas. sp. U2, and phnF of Burkholderia sp. RP007, respectively. The intact DNA fragment was isolated from the chromosomal DNA by Southern hybridization, and expression of nahF gene was investigated on SDS-PAGE.