Ferrate(VI)를 이용한 다양한 수중 환경에서의 tetracycline의 분해 특성 및 반응 경로 연구

박경덕,김일규 대한상하수도학회 2021 상하수도학회지 Vol.35 No.1

Tetracycline is one of the most commonly used as antibiotics for the livestock industry and it is still widely used nowadays. Tetracycline and its metabolites are excreted with excrement, which is difficult to completely removed with conventional sewage treatment, therefore it is apprehended that the tetracycline-resistant bacteria occurs. In this study, the oxidant named ferrate(VI) was used to degrade the tetracycline and investigate the reaction between ferrate(VI) and tetracycline under various aqueous conditions. The highest degradation efficiency of tetracycline occurred in basic condition (pH 10.1 ± 0.1) because of the pKa values of tetracycline and ferrate(VI). The results also showed the effect of water temperature on the degradation of tetracycline was not significant. In addition, the dosage of ferrate(VI) was higher, the degradation of tetracycline and the self-degradation of ferrate(VI) also higher, finally the efficiency of ferrate(VI) was lower. The results said that the various mechanisms effects the reaction of ferrate(VI) oxidation, it required the consideration of the characteristics of the target compound for optimal degradation efficiency. Additionally, intermediate products were detected with LC/MS/MS and three degradation pathways were proposed.

테트라싸이클린 및 테트라싸이클린-구연산 혼합젤로 처리한 치근면의 항 미생물 활성 변화에 관한 연구;In Vivo Study

정희선,한수부,남석우,심창구,계승범,Cheong, Hee-Sun,Han, Soo-Boo,Nam, Seok-Woo,Shim, Chang-Koo,Kye, Seung-Beom 대한치주과학회 1997 Journal of Periodontal & Implant Science Vol.27 No.1

The purpose of this study was to evaluate the substantivity of experimentally developed gel type tetracycline HCl and a mixture of tetracycline-citric acid gel, and compare to those of solution type tetracycline HCl. 11 extracted anterior teeth were subjected to this study. After scaling and root planing teeth were randomly divided into 3 treatments groups : group 1; 3 teeth were irrigated with tetracycline HCl(50mg/ml) solution , group 2; tetracycline gel (5%) was inserted in the periodontal pockets of 3 teeth, group 3; a mixture of tetracycline and citric acid gel was inserted in the pockets of 3 teeth. And 2 teeth treated in 0.9 % sterile saline served as controls. After 5-minute exposure, each tooth immediately extracted and incubated at room temperature for 22 days in tris-buffered saline as a desorption media. The total volume of TBS was removed and replaced with fresh TBS, at 24-h intervals. Removed desorption media transferred to a sterile vial and stored at -70 oC. This procedure was repeated every 24 h throughout the 22-day desorption period. Using Porphyromonas gingivalis as an indicator organism, a microtiter assay was used to evaluate antimicrobial activity desorbed from the teeth. 1. 50mg/ml tetracycline HCl solution exhibited the longest antimicrobial activity. Compared to saline treated group, it showed significant difference on the day 1 and day 2 desorption period. 2. The ODs of 5% tetracycline gel and a mixture of tetracycline-citric acid gel were significantly different during the first 24 hour only. 3. There was no statistically significant difference after the day 3 between the groups.(p<0.05). Despite our expectation a mixture of tetracycline-citric acid gel did not show longer antimicrobial activities than those of tetracycline gel, and the solution type exhibited the longest activities. Because the gel type agents may stay in the subgingival environment longer than the solution, if the teeth were not extracted immediately after the delivery of the agent, the result could be different. hus this result suggests the possibilities of practical use of these kind of gel type agents.

Tetracycline계 항균제에 의한 호중구 Elastase의 효소 활성도 억제 및 그 작용 기전

김우미(Woo Mi Kim),강구일(Koo-Il Kang) 대한약리학회 1993 대한약리학잡지 Vol.29 No.1

Tetracycline계 약제가, 류마치양 관절염을 비롯한 염증성 질환들의 주된 병인으로 알려지고있는 호중구 elastase의 활성도를 억제하였으며, 특히 oxytetracycline, demeclocycline, 그리고 tetracycline 등은 분자 구조적 차이에 따라 elastase의 효소 활성도에 대하여 다양한 억제율을 나타내었다. 측쇄 구조의 5번 위치에 OH^-기가 첨가된 oxytetracycline이 가장 높은 억제율을 나타내었다. 억제 양상에 있어서도 tetracycline이 비경쟁적 저해 형태를 보인 반면에, oxytetracycline은 경쟁적 저해 형태를 나타내었으며, Ki값은 각각 4.9mM과 0.39mM로 산출 되었다. 또한 항균 효과를 나타내는 활성 부위를 제거시킨 de-dimethylaminotetracycline을 합성하여 효소 활성도 억제 실험에 사용한 결과, tetracycline과 유사한 효소 억제 작용을 나타냄을 확인하였다. 이상의 연구 결과에서, tetracycline의 효소 활성도 억제 작용은 항균 효과를 나타내는 활성 부위와 상관없이 독립된 기전에 의해서 일어나는 약리 작용이며, 측쇄 구조의 OH^-기가 이 작용에 영향을 주는 일부 원인인 것으로 추정할 수 있으며, 이를 tetracycline계 약제가 염증 부위에서 나타내는 분자 단계에서의 새로운 약리 기전으로 제시하고자 한다. 또한 de-dimethylaminotetracycline은 항균제의 장기 사용시에 발생할 수 있는 저항균의 출현과는 무관하므로, 다른 부작용에 대한 연구가 선행될 경우, elastase에 의해 야기되는 만성 질환들의 치료제로써 중요한 역할을 할 것으로 사료된다. Human neutrophil elastase (HNE, EC 3,4,21, 11), a mediator of tissue breakdown, was inhibited by tetracycline, oxytetracycline and demeclocycline. Among them, oxytetracycline showed the most potent inhibitory effect on the activity of HNE. IC50 of this drug at our specific condition was less than 1 mM. Tetracycline inhibited human neutrophil elastase non-competitively, and oxytetracycline inhibited competitively. Ki values of tetracycline and oxytetracycline were 4.9 mM and 0.39 mM, respectively. Structural modified tetracycline, de-dimethylaminotetracycline, which showed no antibiotic activity since the active dimethylamino radical was removed from the position #4 of the tetracycline, showed similar inhibition effect on the activity of human neutrophil elastase to that of tetracycline. Thus, we speculated that inhibition of human neutrophil elastase by tetracyclines was not depended on the dimethylamino radical which is a critical active site for antibiotic effect, rather it was depended on the hydoxyl radical of tetracyclines. Therefore, the property of inhibiting elastase may be an additional molecular biochemical mechanism of action of these drugs at the inflammatory sites.

Inhibition of Human Neutrophil Elastase by Tetracyclines and Mechanism of the Inhibition

김우미,강구일,Kim, Woo-Mi,Kang, Koo-Il The Korean Society of Pharmacology 1993 대한약리학잡지 Vol.29 No.1

Tetracycline계 약제가, 류마치양 관절염을 비롯한 염증성 질환들의 주된 병인으로 알려지고있는 호중구 elastase의 활성도를 억제하였으며, 특히 oxytetracycline, demeclocycline, 그리고 tetracycline 등은 분자 구조적 차이에 따라 elastase의 효소 활성도에 대하여 다양한 억제율을 나타내었다. 측쇄 구조의 5번 위치에 $OH{^-}$기가 첨가된 oxytetracycline이 가장 높은 억제율을 나타내었다. 억제 양상에 있어서도 tetracycline이 비경쟁적 저해 형태를 보인 반면에, oxytetracycline은 경쟁적 저해 형태를 나타내었으며, Ki값은 각각 4.9mM과 0.39mM로 산출 되었다. 또한 항균 효과를 나타내는 활성 부위를 제거시킨 de-dimethylaminotetracycline을 합성하여 효소 활성도 억제 실험에 사용한 결과, tetracycline과 유사한 효소 억제 작용을 나타냄을 확인하였다. 이상의 연구 결과에서, tetracycline의 효소 활성도 억제 작용은 항균 효과를 나타내는 활성 부위와 상관없이 독립된 기전에 의해서 일어나는 약리 작용이며, 측쇄 구조의 $OH{^-}$기가 이 작용에 영향을 주는 일부 원인인 것으로 추정할 수 있으며, 이를 tetracycline계 약제가 염증 부위에서 나타내는 분자 단계에서의 새로운 약리 기전으로 제시하고자 한다. 또한 de-dimethylaminotetracycline은 항균제의 장기 사용시에 발생할 수 있는 저항균의 출현과는 무관하므로, 다른 부작용에 대한 연구가 선행될 경우, elastase에 의해 야기되는 만성 질환들의 치료제로써 중요한 역할을 할 것으로 사료된다. Human neutrophil elastase (HNE, EC 3,4,21, 11), a mediator of tissue breakdown, was inhibited by tetracycline, oxytetracycline and demeclocycline. Among them, oxytetracycline showed the most potent inhibitory effect on the activity of HNE. IC50 of this drug at our specific condition was less than 1 mM. Tetracycline inhibited human neutrophil elastase non-competitively, and oxytetracycline inhibited competitively. Ki values of tetracycline and oxytetracycline were 4.9 mM and 0.39 mM, respectively. Structural modified tetracycline, de-dimethylaminotetracycline, which showed no antibiotic activity since the active dimethylamino radical was removed from the position #4 of the tetracycline, showed similar inhibition effect on the activity of human neutrophil elastase to that of tetracycline. Thus, we speculated that inhibition of human neutrophil elastase by tetracyclines was not depended on the dimethylamino radical which is a critical active site for antibiotic effect, rather it was depended on the hydoxyl radical of tetracyclines. Therefore, the property of inhibiting elastase may be an additional molecular biochemical mechanism of action of these drugs at the inflammatory sites.

Hep3B 세포주에서 tetracycline에 의한 p21 의 발현

강경화,최경희 中央大學校 遺傳工學硏究所 1997 遺傳工學硏究論集 Vol.10 No.1

본 연구에서는 p21의 발현이 조절되는 세포주를 만들고자 사람의 간암 세포주인 Hep3B 세포에 tetracycline-repressible vector와 tetracycline-inducible vector를 형질전환 시킨 후 네오마이신 선택 배지에서 배양하여 얻은 클론을 pUHD10-3/p21 vector에 특이적인 PCR primer를 사용하여 genomic DNA PCR을 실시하였다. 이렇게 해서 얻어진 클론중에서 p21의 발현이 높은 클론을 찾기 위해 p21에 대한 western blotting을 실시하여 p21의 단백질 수준에서의 발현이 높은 클론을 선택하였다. 마지막으로 tetracycline에 의해 p21의 발현이 조절되는지를 조사한 결과 15C31 클론에서 tetracycline을 제거하고 48시간 동안 배양한 후에는 7㎍/ml의 tetracycline을 첨가했을 때에 비해 약 2배정도 높게 발현됨을 알 수 있었다. 또한 본 연구에서는 tetracycline-repressible vector system이 tetracycline-inducible vector system보다 더 효과적임을 알 수 있었다. p21 is known as a cyclin-dependent kinase inhibitor and participated in cell cycle arrest. apoptosis and differentiation. To directly demonstrate the role of p21. we have expressed p21 cDNA in human hepatocarcinoma cell line. Hep3B. using tetracycline repressible system and tetracycline inducible system. We obtained 8 clones as a tetracycline repressible vector 7 clones as a tetracycline inducible vector by using neomycin selection and genomic DNA PCR method. A dose dependent inhibition of p21 expression by tetracycline was observed in tetracycline repressible system. p21 expression was maximal in the absence of tetracycline and the expression level was 2 fold higher than that of p21 in the cell culture with 7㎍/ml of tetracycline.

닭에서 분리한 장내세균의 테트라사이클린 내성 유전자의 분포

박인달,배일권 고신대학교의과대학 2008 고신대학교 의과대학 학술지 Vol.23 No.4

Background : Widespread resistance to the broad-spectrum tetracyclines has been caused by heavy clinical use and misuse in the human population. Additional contributing factors are the use of tetracyclines as a growth promotor in agriculture and as infection control agent in domestic animals and aquaculture. The purpose of this study was to investigate susceptibility of tetracycline and to detect tetracycline resistant genes from chickens. Methods : Thirty-seven tetracycline resistant enteric bacteria were collected from the rectal swab of chickens by disk diffusion method, and the MICs (minimal inhibitory concentration) of tetracycline, oxytetracycline, chlortetracycline and erythromycin were determined by agar dilution method. PCR screening was carried out to identify possible tet genes that contributed to tetracycline resistance. Results : Of the 37 tetracycline-resistant isolates, the frequency of isolation was Escherichia coli 32 (84.2%), Citrobacter freundii 2 (5.3%), Klebsiella pneumoniae 1 (2.5%), Lectercia adecarboxylata 1 (2.5%), Proteus vulgaris 1 (2.5%). The MIC range for the tetracycline-resistant enteric bacteria was as follows: tetracycline, >256 ㎍/㎖; oxytetracycline, >256 ㎍/㎖; chlortetracycline, 16∼>128 ㎍/㎖; erythromycin, 32∼>256 ㎍/㎖. All of the 37 tetracycline-resistant isolates were positive for tet(A) gene (100%), and 8 (21.6%) of these isolates were found to harbor the tet(A) plus tet(B) genes. Conclusion : All isolates were resistant to tetracycline, oxytetracycline, chlortetracycline and erythromycin, and they harbored tet(A) gene or tet(A) plus tet(B) genes that gave rise to the tetracycline resistance. These results show that enteric bacteria isolates from chicken in Korea are resistant to tetracycline and all of them harbored tet(A) gene.

테트라싸이크린 함유 calcium sulfate의 서방형 국소 약물 송달 효과에 대한 연구

김성희,최성호,조규성,채중규,박광균,김종관,Kim, Sung-Hee,Choi, Seong-Ho,Cho, Kyoo-Sung,Chai, Jung-Kiu,Park, Kwang-Kyun,Kim, Chong-Kwan 대한치주과학회 1997 Journal of Periodontal & Implant Science Vol.27 No.4

치주질환은 세균에 의한 감염성 질환으로, 기계적 치태제거에 의한 치료의 한계를 극복하기 위하여 항세균 제재를 통한 화학적 치태 및 세균제거가 필요하게 되었다. 전신적으로 항생제를 투여할 경우 유효농도의 유지를 위해 많은 양의 약물이 투여되어야 하고, 여러가지 부작용의 위험이 있으므로, 이러한 단점을 극복하기 위한 국소 약물 송달체계가 필요하게 되었다. 본 실험의 목적은 치주질환 치료에 가장 많이 쓰이는 tetracycline을 calcium sulfate와 혼합하여, calcium sulfate의 서방형 국소 약물 송달효과에 대해 알아보고자 함이다. Modified calcium sulfate paste 와 10% tetracycline을 혼합한 것을 실험 1군으로, calcium sulfate와 10% tetracycline을 혼합하여 완전 경화 시킨 것을 실험 2군으로, calcium sulfate와 10% tetracycline을 혼합하여 경화되기 전에 사용한 것을 실험 3군으로, tetracycline-ethylene vinyl acetate fiber를 실험 4군으로 하여 시간별 tetracycline 방출농도, 유효농도 지속시간, calcium sulfate의 흡수기간 등을 관찰하여 다음과 같은 결론을 얻었다. 1. 실험 1군은 실험 5주까지 유효농도($4{\mu}g/ml$)이상의 농도를 유지하였고, 실험 2군은 실험 9일까지 유지하였으며, 실험 3군은 실험 7일까지 유효농도이상의 농도를 유지하였고, 실험 4군은 실험 15일까지 유지하였다. 2. 실험 2군은 평균 11.8일에 완전 용해 되었고, 실험 3군은 14.8일에 완전용해되었다. 3. 실험 2군과 3군은 실험 1주까지 방출된 tetracycline의 농도에 유의차를 보이지 않았다(p<0.05). 이상의 결과에서 볼 때, calcium sulfate는 tetracycline과 혼합하였을 때 일정 기간 동안 충분한 양의 tetracycline을 방출시키고, $11{\sim}14$일 정도 후에는 완전히 녹아 없어지므로, 서방형 국소 약물 송달 제재로서 가능성이 있으리라 생각된다. Periodontal disease is a bacterially causal by disease, To remove plaque and bacteria, it has been necessary to prescribe chemical drug to patient to subjugate therapeutic unvalue by mechanical scaling. As a patient on a high dosage of the antibiotics to maintain the effective concentration may produce unfavorable side effects, this decase demands the Slow-release local drug delivery system. The object of the experiment is to study on the slow-release local drug delivery effects of calcium sulfate compounded with tetracycline that mainly used in periodontal disease. Experimental groups were divided into four classes as follow: Group 1 10% tetracycline compounded modified calcium sulfate paste. Group 2 : compounded and hardened 10% tetracycline and calcium sulfate. Group 3 : compounded 10% tetracycline and calcium sulfate, used Just before hardened. Group 4 : tetracycline-ethylene vinyl acetate fiber. In the four groups, release concentration, it's durability and the period of absorption by times are observed and concluded as follow: 1. An effective concentration($4{\mu}g/ml$) remained until 5 weeks in group 1, 9 days in group 2, 7 days in group 3, 15 days in group 4. 2. It was fully fused at 11.8 days average in group 2 and 14.8 days average in group 3. . There were no statistically significant results in tetracycline concentration until a week in group 2 and 3(p<0.05) These results suggest that tetracycline loaded calcium sulfate release sufficient tetracycline and fused in $11{\sim}14$ days, so calcium sulfate is useful carrier as slow release local drug delivery system.

활성탄 재질 및 사용연수에 따른 Tetracycline계 항생물질 흡착특성

손희종(Hee Jong Son),정종문(Jong Moon Jung),황영도(Young Do Hwang),노재순(Jae Soon Roh),유평종(Pyung Jong Yu) 大韓環境工學會 2008 대한환경공학회지 Vol.30 No.9

입상활성탄 재질별 신탄 및 사용탄에서의 tetracycline계 항생물질 4종에 대한 파과특성의 경우 석탄계 활성탄이 가장 늦게 파과에 도달하였으며, 다음으로 야자계, 목탄계 순으로 조사되었다. 또한, 물질별 활성탄에서의 파과특성을 살펴보면 tetracycline(TC)의 파과시점이 가장 늦은 것으로 나타났으며, 다음으로 oxytetracycline(OTC), chlortetracycline(CTC), minocycline(MNC)으로 나타났다. 활성탄 g당 tetracycline계 항생물질 4종에 대한 최대 흡착량(X/M)은 석탄계 활성탄이 가장 높은 것으로 나타났으며, 다음으로 야자계와 목탄계 순으로 나타났다. tetracycline계 항생물질 4종에 대한 석탄계 활성탄의 최대 흡착량(X/M)은 야자계와 목탄계 활성탄에 비해 각각 1.27∼1.36배 및 1.69∼1.84배 정도 높은 것으로 조사되었다. 활성탄 사용율(carbon usage rate, CUR)은 tetracycline의 경우 석탄계 재질의 활성탄이 2.96 g/일, 야자계나 목탄계 활성탄은 각각 3.40 g/day 및 4.53 g/day의 활성탄을 사용하여야만 제어가 가능한 것으로 조사되어 석탄계 활성탄이 다른 재질의 활성탄들에 비해 적은 양으로도 tetracycline계 항생물질을 제어할 수 있는 것으로 나타났으며, 나머지 tetracycline계 항생물질 3종에서도 이와 유사한 결과를 나타내었다. 또한, 석탄계 활성탄 신탄과 사용탄에 대한 CUR을 비교해보면 tetracycline의 경우 신탄을 사용하였을 경우 보다 1.3년 사용탄 및 3.1년 사용탄을 사용하였을 경우가 CUR이 1.96배 및 2.53배 정도 높은 것으로 나타났다. Adsorption performance of tetracycline antibiotic compounds such as tetracycline(TC), oxytetracycline(OTC), chlortetracycline(CTC) and minocycline(MNC) on granular activated carbon(GAC) was evaluated in this study. The coal-based activated carbon was found to be more effective than other carbons in adsorption of tetracycline antibiotic compounds. The wood-based activated carbon was less effective than coconut- and coal-based carbon in adsorption nevertheless having larger pore volume and specific surface area than others carbons. The maximum adsorption capacities(X/M) of coal-based activated carbon for the four tetracycline species was 1.27∼1.36 and 1.69∼1.84 times larger than coconut- and wood-based activated carbon, respectively. Carbon usage rates(CUR) of coal-, coconut- and wood-based activated carbons for tetracycline(TC) were 2.96 g/day, 3.40 g/day and 4.53 g/day, respectively. Similar results were obtained in the adsorption of the rest three tetracycline species. It is concluded that coal-based activated carbon could removed the tetracycline antibiotic compounds better than other material-based activated carbons.

Removal of the antibiotic tetracycline by Fe-impregnated SBA-15

Eun Woo Shin,Bao Khanh Vu,Olga Snisarenko,Wang Seok Jeong,Hak Sung Lee 한국화학공학회 2010 Korean Journal of Chemical Engineering Vol.27 No.1

We prepared Fe-impregnated mesoporous silicates to investigate the adsorption of tetracycline in aqueous solution. Mesoporous silicates with different Fe content (5, 10, 30, 50 wt%) were prepared by an incipient wetness method. Adsorption kinetics for tetracycline showed that Fe-impregnation improved the adsorption ability for tetracycline. By fitting the adsorption kinetic data to a pseudo second-order model, we obtained a maximum adsorption amount of tetracycline with Fe30SBA-15 (30 wt% Fe-impregnated mesoporous silicates) at 41.7 mg/g. The pH dependency of tetracycline adsorption exhibited a volcano curve where the maximum adsorption onto the Fe30SBA-15 sample occurred in the neutral pH region. The introduction of Fe species into the SBA-15 revived the adsorption ability for tetracycline,whereas there was no interaction between tetracycline and SBA-15, a mesoporous silicate. These results suggest that impregnated Fe species produce an effective interaction with tetracycline in an aqueous system.

Genetic diversity of the tet(M) gene in tetracycline-resistant oral streptococci

( Dasul Jeong ),( Si Young Lee ) 조선대학교 치의학연구원(구 조선대학교 구강생물학연구소) 2021 Oral Biology Research (Oral Biol Res) Vol.45 No.2

Tetracycline resistance occurs at a high frequency among clinical isolates of both gram-positive and gram-negative bacteria. The mechanism and genetics of tetracycline resistance have not been extensively studied in streptococci, although the overwhelming majority of clinical isolates are tetracycline resistant. tet (M) is the most common tetracycline-resistance gene in streptococci. The aim of this study was to examine the genetic diversity of tet (M) genes in tetracycline-resistant oral streptococci from dental plaque. Streptococci were isolated from supragingival plaque samples of healthy persons. The isolates were then identified at the species level, and the minimum inhibitory concentration (MIC) of tetracycline was determined. Genomic DNA was extracted from tetracyclineresistant isolates and tet (M) was amplified using polymerase chain reaction with tet (M)-specific primers. The polymerase chain reaction products were cloned, DNA sequencing was performed, and the sequences were compared using an alignment program. The estimated nucleotide divergence between different tet (M) alleles ranged from 0.00% to 6.07% among oral streptococci. The tet(M) genes from oral streptococci consisted of regions similar in sequence, interspersed with regions that differed at some nucleotide sites, revealing a mosaic structure. The percent nucleotide divergence of tet(M) was unrelated to the MIC values of tetracycline for oral streptococci, and bacterial strains in the same streptococcal species showed different heterogeneity in tet(M). The divergences of tet(M) nucleotide sequences among oral streptococci were comparable with those of other bacterial genera. Our findings may provide basic information about the transposition processes associated with tet(M) in oral streptococci.