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Ototoxic destruction by co-administration of kanamycin and ethacrynic acid in rats

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2011年第10期12卷 853-861页

作者： Hong LIU Da-lian D1NG Hai-yan JIANG Xue-wen WU Richard SALVI Hong SUN Center for Hearing and Deafness University at Buffalo State University of New York New York 14214 USA Department of Otolaryngology the Third Xiangya Hospital of Central South University Changsha 410013 China Department of Otolaryngology Xiangya Hospital of Central South University Changsha 410008 China

It is well known that ethacrynic acid (EA) can potentiate the ototoxicity of aminoglycoside antibiotics (AmAn) such as kanamycin (KM),if they were applied at the same time.Currently,to create the model of EA-KMinduced cochlear lesion in rats,adult rats received a single injection of EA (75 mg/kg,intravenous injection),or followed immediately by KM (500 mg/kg,intramuscular injection).The hearing function was assessed by auditory brainstem response (ABR) measurement in response to click and/or tone bursts at 4,8,12,16,20,24,and 32 kHz.The static microcirculation status in the stria vascularis after a single EA injection was evaluated with eosin staining.The pathological changes in cochlear and vestibular hair cells were also quantified after co-administration of EA and KM.After a single EA injection,blood flow in vessels supplying the stria vascularis rapidly diminished.However,the blood supply to the cochlear lateral wall partially recovered 5 h after EA treatment.Threshold changes in ABR were basically parallel to the microcirculation changes in stria vascularis after single EA treatment.Importantly,disposable co-administration of EA and KM resulted in a permanent hearing loss and severe damage to the cochlear hair cells,but spared the vestibular hair cells.Since the cochlear lateral wall is the important part of the blood-cochlea barrier,EA-induced anoxic damage to the epithelium of stria vascularis may enhance the entry of KM to the cochlea.Thus,experimental animal model of selective cochlear damage with normal vestibular systems can be reliably created through co-administration of EA and KM.

关键词： Ototoxicity Ethacrynic 酸 kanamycin 老鼠血蜗障碍

The effect of Some Boron Derivatives on kanamycin Resistance and Survival of E.coli and P.aeruginosa in Lake Water

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Biomedical and Environmental Sciences 2012年第4期25卷 476-482页

作者： Cihan DARCAN Mustafa KAHYAOGLU Department of Biology Faculty of Art and SciencesDumlupinar University Faculty of Education Siirt University

Objective To study MIC value of 7 boron derivatives namely [Boric acid （H3BO3）, Anhydrous Borax （Na2B407）, Sodium Borate （NaBO2）, Diammonium Tetraborate （NH4）2B4O7, Sodium Perborate （NaBO3）, Boron Trioxide （B203）, Potassium Tetraborate （K2B407）] on E. coil and P. aeruginosa and their effects on survival of bacteria in lake water and resistance against kanamycin antibiotic. Methods MIC values of Boron derivatives and antibiotic were studied by broth microdilution method. The effect of boron derivatives on survival of bacteria in lake water were also determined with plate count. Results Sodium perborate was determined as the substances. Effectiveness increased as temperature most effective substance among the studied increased. E. coil was more affected from P. aeruginosa in 8 mg/mL sodium perborate concentration in lake water. Moreover, it was determined that MIC value of kanamycin antibiotic decreased 200 times by especially treating P. aeruginosa with sodium perborate in lake water. However, it can be stated that this change in resistance did not arise from microorganisms. Conclusion Sodium perborate solution can be used supportedly in kanamycin antibiotic applications for P. aeruginosa. Future studies are necessary to explore the relation between sodium perborate and kanamycin which is effective on P. aeruginosa in lake water.

关键词： Boron Lake water kanamycin E. coli P. aeruginosa

Graphene-Gold Nanoparticle-modified Electrochemical Sensor for Detection of kanamycin Based on Target-induced Aptamer Displacement

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Chemical Research in Chinese Universities 2018年第6期34卷 952-958页

作者： ZHENG Jingyi FENG Rongrong HE Caimei LI Xiaoxia School of Chemistry and Chemical Engineering Yah'an University Yah'an 716000 P. R. China

A highly sensitive and selective label-free electrochemical sensor was developed for the determination of kanamycin. To improve the sensitivity of the electrochemical sensor, graphene-gold nanoparticles were prepared by a one-step electrochemical coreduction process and were modified on the surface of a glassy carbon electrode. The double-stranded DNA（ds-DNA） duplex probe was immobilized onto the graphene-gold nanoparticle-modified elec-trode. The introduction of target kanamycin induced the displacement of aptamer from the ds-DNA duplex into the solution. Methylene blue（MB） as a redox indicator monitored the current change using differential pulse voltammetry Under optimal conditions, the designed electrochemical aptasensor exhibited a wide linear range from 0. i pmol/L to 10 pmol/L with a detection limit of 0.03 pmol/L for kanamycin. The experimental strategy enabled the direct analysis of milk samples, and the results showed high sensitivity and good selectivity.

关键词： kanamycin Electrochemical aptasensor Graphene Gold nanoparticle Methylene blue

A Label-free and Functional Fluorescent Oligonucleotide Probe Based on a G-Quadruplex Molecular Beacon for the Detection of kanamycin

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Chemical Research in Chinese Universities 2018年第4期34卷 541-545页

作者： ZHU Yuqing LI Wei TAN Suzhen CHEN Tianxiao School of Chemistry and Biological Engineering Changsha University of Science and Technology Changsha 410004 P. R. China

A label-free and turn-off fluorescent method for the quantitative detection of kanamycin based on a funtional molecular beacon was developed. The molecular beacon consists of two hairpin structures with a split G-rich oligonucleotide in the middle. The kanamycin＇s aptamer formed the loops portion for recognizing kanamycin, and the G-quadruplex bound by Thioflavin T（ThT） was employed as the reporter. In the absence of target, the molecular beacon folded into double stem-loops and the splited G-rich oligonucleotid came close to form a G-quadruplex. When ThT bound to the G-quadruplex, the fluorescence intensity of the solution increased. Upon the addition of kanamycin, the function between kanamycin and aptamer unfolded the hairpin and disassembled the G-quadraplex structure, resulting in a significant decrease in the fluorescence intensity. A good linear relationship ranging from 0.7 nmol/L to 10 nmol/L was achieved and the limit of detection was 0.37 nmol/L. Besides, it could efficiently recognize kanamycin in real samples.

关键词： Molecular beacon Fluorescence kanamycin G-Quadruplex Thioflavin T

Preparation of Polyclonal Antibody and Development of a Biotin-streptavidin-based ELISA Method for Detecting kanamycin in Milk and Honey

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Chemical Research in Chinese Universities 2017年第6期33卷 876-881页

作者： SU Ping CHEN Xiaonan HE Zhangjing YANG Yi Beijing Key Laboratory of Environmentally Harmful Chemical Analysis College of Science Beijing University of Chemical Technology Beijing 100029 P. R. China

kanamycin is an aminoglycoside antibiotic used increasingly in human and veterinary medicine. However, kanamycin residues in food can cause serious side effects, Here we reported the preparation of polyclonal antibody and the development of an indirect competitive biotin-streptavidin-amplified-based enzyme-linked immunosorbent assay（BA-ELISA） that can sensitively and specifically detect kanamycin residues in milk and honey. The immtmogen and coating antigen were synthesized by covalently linking kanamycin to carrier proteins using the carbodiimide method. The anti-kanamycin polyclonal antibodies were obtained from immunized rabbits. The key assay parameters were investigated and optimized. The results show that under optimum conditions, the limit of detection for kanamycin is 0.07 ng/mL and the ICs0 is 6.48 ng/mL. Cross-reactivity values of the antibody with four kanamycin analogues are all 〈1%. Trace amounts of kanamycin in milk and honey samples can be detected by this novel BA-ELISA method successfully with satisfactory recoveries of 91.0%-103.3%. The developed protocol was also validated against liquid chromatography-mass spectrometry, returning a significant correlation. These results indicate that BA-ELISA is a viable option for monitoring kanamycin residues in milk and honey.

关键词： kanamycin Biotin-streptavidin-amplified system Enzyme-linked immtmosorbent assay（ELISA） Milk Honey

Microcalorimetric Evaluation of the Effect of kanamycin： An Analysis Based on the Median-Effect Principle

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Chinese Journal of Chemistry 2007年第1期25卷 20-24页

作者：王莉衡范代娣尚龙安施惠娟马晓轩米钰顾利锋徐抗震 Department of Chemical Engineering Northwest University/Shaanxi Source and Application Chemical Engineering Center Xi'an Shaanxi 710069 China Key Laboratory of Resource Biology and Biotechnology in Western China (Northwest University) Ministry of Education Xi'an Shaanxi 710069 China Collage of Biological and Chemical Engineering Ningbo Institute of Technology Zhejiang Univerisity Ningbo Zhejiang 315100 China Department of Reproductive Biology Shanghai Institute of Planned Parenthood ResearchShanghai 200032 China

A study of the effect of drug, kanamycin, on the growth metabolism of recombinant Escherichia coli B 1 was carded out by microcalorimeter monitoring of the metabolic activity of treated cells. Power-time curves of growing recombinant Escherichia coli cell suspensions, treated with different kanamycin doses, were recorded. The extent of the effect was evaluated by changes in the slopes of the microcalorimetric curves and the kinetics of the drug action was interpreted from the time at which these changes reached their maximum values and maintained their maximum values. Experimental dose-effect relationships conform to the median-effect principle of the mass-action law： fa/（1-fa）=（D/D50）^m. A plot of y=lg[（fa）^1-1]^-1 versus x=lg D gives the slope m, D50 and R∞. The experimental results revealed that high concentration of kanamycin had an inhibitory effect on the growth of recombinant Escherichia coli B 1 in the lg phase, and had a promoting effect in the stationary period. Moreover, it was demonstrated that microcalorimetry was a reliable method for the detection of modulatory effects in biology.

关键词： kanamycin microcalorimetry recombinant Escherichia coli

Effects of combined administration of furosemide and kanamycin on rat auditory nerve

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Journal of Otology 2011年第1期6卷 26-30页

作者： GUO Weiwei YUAN Fen-qian LIU Hui-zhan YANG Shi-ming Department of Otolaryngology Head ＆ Neck Surgery Institute of Otolaryngology Chinese PLA General Hospital Beijing 100853 P.R. China Department of Head and Neck Surgery Jiangxi Cancer Hospital Nanchang 330029 China

Objective To determine the effects of combined administration of furosemide and kanamycin on inner ear structures and the auditory nerve in rats. Methods The rats in the treatment group received intravenous injections of combined furosemide and kanamycin sulfate, and the rats in the normal control group received no treatment. The auditory brainstem response （ABR） test was carried out 7 days after drug administration to determine the effects of drug administration on hearing. Cochlear slice and cochlear wholcmount were prepared after 7 days of drug treatment. Results After 7 days of drug administration, ABR thresholds were significantly higher in the treatment group than in the control group and neurofilaments were significantly reduced, although the number of spiral ganglia showed no decrease and there were no signs of supporting cell injury. Conclusions Combined administration of furosemide and kanamyein sulfate has an apparent synergistic ototoxic effect. Although spiral ganglion damage may not be apparent within a short time period of drug administration, damage to auditory nerve fibers is obvious.

关键词： kanamycin furosemide deafness model hair cell injury

Aminoglycoside ototoxicity in three murine strains and effects on NKCC1 of stria vascularis

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Chinese Medical Journal 2006年第12期119卷 980-985页

作者： CHU Han-qi XIONG Hao Zhou Xiao-qin HAN Fang WU Zhen-gong ZHANG Ping HUANG Xiao-wen CUl Yong-hua Department of Otolaryngology Tongji Hospital Tongji MedicalCollege Huazhong University of Science and Technology Wuhan430030 China Union Hospital Tongji Medical College Huazhong University ofScience and Technology Wuhan 430022 China

Background After establishing a murine model of aminoglycoside antibiotic （AmAn） induced ototoxicity, the sensitivity of AmAn induced ototoxicity in three murine strains and the effect of kanamycin on the expression of Na-K-2Cl cotransporter-1 （NKCC1） in stria vascularis were investigated. Methods C57BL/6J, CBA/CaJ, NKCC1^＋/- mice （24 of each strain） were randomly divided into four experimental groups： A： kanamycin alone; B： kanamycin plus 2,3-dihydroxybenzoate; C： 2,3-dihydroxybenzoate alone; and D： control group. Mice were injected with kanamycin or/and 2,3-dihydroxybenzoate twice daily for 14 days. Auditory brainstem response （ABR） was measured and morphology of cochlea delineated with succinate dehydrogenase staining. Expression of NKCC1 in stria vascularis was detected immunohistochemically. Results All three strains in groups A and B developed significant ABR threshold shifts （P〈0.01）, which were accompanied by outer hair cell loss. NKCC 1 expression in stria vascularis was the weakest in group A （A cf D, P〈0.01） and the strongest in groups C and D （P〈0.05）. CBA/CaJ mice had the highest sensitivity to AmAn.Conclusions Administration of kanamycin established AmAn induced ototoxicity. kanamycin inhibited the expression of NKCC1 in stria vascularis. 2, 3-dihydroxybenzoate attenuated AmAn induced ototoxicity-possibly by enhancing the expression of NKCC 1. Age related hearing loss did not show additional sensitivity to AmAn induced ototoxicity in murine model.

关键词： kanamycin 2,3-dihydroxybenzoate ototoxicity reactive oxygen species hearing loss

PCR Targeting of Antibiotic Resistant Bacteria in Public Drinking Water of Lahore Metropolitan,Pakistan

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Biomedical and Environmental Sciences 2009年第6期22卷 458-463页

作者： ZAHOOR QADIR SAMRA MARIAM NASEEM SUMARIA JAVED KHAN NADIA DAR MUHAMMAD AMIN ATHAR Institute of Biochemistry and Biotechnology Quaid-i-Azam CampusUniversity of the PunjabLahore 54590Pakistan Jinnah Degree College for Women Mozang RoadLahorePakistan

Objective To investigate the prevalence of kanamycin （kan） and ampicillin （amp） resistant bacteria in public drinking water. Methods Bacteria containing kan and amp resistant genes were amplified by PCR and further characterized by colony hybridization and transformation studies. The genus of kan and amp resistant bacteria was determined with standard methods. Results Among the 625 drinking water samples, 400 contained kan and amp resistant bacteria and the percentage was 42.5% and 57.5%, respectively, which was further confirmed by the amplification of a 810 bp kan resistant gene and a 850 bp amp resistant gene. Of the 170 kan resistant bacteria, 90 were Gram negative and 80 were Gram positive. Of the 230 amp resistant bacteria, 160 were Gram negative while 70 were Gram positive. Salmonella, Shigella, Staphylococcus, Streptococcus, and E.coli were detected as 13%, 11%, 17%, 30%, and 29%, respectively. Bacterial strain DH5α transformed with plasmids isolated from kan and amp resistant bacteria confirmed that the antibiotic resistant genes were mediated by plasmids. Conclusion Drinking water is contaminated with kan and amp resistant bacteria due to poor sanitary conditions.

关键词： kanamycin Ampicillin PCR Plasmid Colony hybridization