WorldWideScience

Sample records for kanamycin neomycin polymixin

  1. Inactivation of kanamycin, neomycin, and streptomycin by enzymes obtained in cells of Pseudomonas aeruginoa.

    Science.gov (United States)

    Doi, O; Ogura, M; Tanaka, N; Umezawa, H

    1968-09-01

    Ten strains of Pseudomonas aeruginosa were disrupted and centrifuged. The supernatant fluids from centrifugation at 105,000 x g contained enzymes inactivating kanamycin, neomycin, and streptomycin in the presence of adenosine triphosphate. Kanamycin-inactivating enzyme was precipitated with ammonium sulfate at 66% of saturated concentration, and the inactivated kanamycin was shown to be kanamycin-3'-phosphate in which the C-3 hydroxyl group of 6-amino-6-deoxy-d-glucose moiety was phosphorylated. This is identical with kanamycin inactivated by Escherichia coli carrying R factor. Streptomycin-inactivating enzyme was precipitated with ammonium sulfate at 33% of saturated concentration.

  2. Impairment of membrane phosphoinositide metabolism by aminoglycoside antibiotics: streptomycin, amikacin, kanamycin, dibekacin, gentamicin and neomycin.

    Science.gov (United States)

    Marche, P; Koutouzov, S; Girard, A

    1983-11-01

    Like many amphiphilic cationic drugs, aminoglycosides are able to produce phospholipidosis, mainly by inhibiting enzymes involved in phospholipid metabolism. Phosphoinositides have been suggested to function as receptors for aminoglycosides. Therefore, we investigated the influence of these drugs upon phosphoinositide metabolism by measuring the 32P-incorporation into the polyphosphoinositides, using the rat erythrocyte membrane as a model. Depending upon the experimental conditions, neomycin induced a decrease and/or an increase in the 32P-labeling of triphosphoinositides (TPI) and of diphosphoinositides (DPI), respectively. These variations were rapid and depended upon the drug concentration. At 0.3 mM, neomycin reversed the distribution of radioactivities associated with DPI and TPI without modifying the total radioactivity incorporated. This drug concentration altered neither the Mg++-activated TPI-specific phosphomonoesterase activity nor the Ca++-activated polyphosphoinositide phosphodiesterase activity. It appears likely that the drug inhibits the DPI-kinase activity, by interacting with DPI and thereby lowering the substrate availability. Over the range of concentrations studied (up to 1-2 mM), gentamicin, kanamycin and dibekacin behave as neomycin. However, their effects could be observed only at drug concentrations higher than those of neomycin. By contrast, streptomycin and amikacin did not alter the 32P-labeling of TPI and of DPI. The order of potency of aminoglycosides for the impairment of the phosphoinositide interconversion was neomycin, gentamicin, dibekacin, kanamycin. A possible relationship between the toxicity of aminoglycosides and their capacity to impair the phosphoinositide metabolism is discussed.

  3. Neamin as an immunogen for the development of a generic ELISA detecting gentamicin, kanamycin and neomycin in milk

    NARCIS (Netherlands)

    Loomans, E.E.M.G.; Wiltenburg, van J.; Koets, M.; Amerongen, van A.

    2003-01-01

    A broad-specific ELISA using one antibody preparation for the detection of gentamicin, kanamycin, and neomycin in milk is reported for the first time. For the immunization of rabbits, neamin was used as the generic hapten on the basis of the facts that it is a two-ring fragment of neomycin and, in s

  4. Neamin as an immunogen for the development of a generic ELISA detecting gentamicin, kanamycin and neomycin in milk

    NARCIS (Netherlands)

    Loomans, E.E.M.G.; Wiltenburg, van J.; Koets, M.; Amerongen, van A.

    2003-01-01

    A broad-specific ELISA using one antibody preparation for the detection of gentamicin, kanamycin, and neomycin in milk is reported for the first time. For the immunization of rabbits, neamin was used as the generic hapten on the basis of the facts that it is a two-ring fragment of neomycin and, in s

  5. Resistance mechanisms of kanamycin-, neomycin-, and streptomycin-producing streptomycetes to aminoglycoside antibiotics.

    Science.gov (United States)

    Hotta, K; Yamamoto, H; Okami, Y; Umezawa, H

    1981-09-01

    Streptomyces kanamyceticus ISP5500, S. fradiae ISP5063 and S. griseus ISP5236, which produce kanamycin, neomycin or streptomycin respectively, were highly resistant to the antibiotics they produced. Polyphenylalanine synthesis in cell free systems was also resistant to the action of the antibiotics. Reciprocal exchange between ribosomes and S150 fractions from the three strains revealed that the S150 fraction of each strain had an enzyme activity that inactivated the appropriate antibiotic whereas the ribosomes were susceptible to the antibiotics. It was concluded that the resistance of the in vitro polyphenylalanine synthesizing systems of these antibiotics was due to the presence of inactivating enzymes. Furthermore, S. fradiae and S. kanamyceticus were highly resistant to aminocyclitol-containing aminoglycoside antibiotics other than those produced by the two strains. In these cases, the inactivating enzymes were found to have a major role in the resistance mechanism. However, the resistance of S. kanamyceticus ISP5500 to streptomycin seems to be due to resistance at the ribosomal level.

  6. Antibiotic radioprotection of mice exposed to supralethal whole-body irradiation independent of antibacterial activity. [Gamma radiation, streptomycin, kanamycin, neomycin, gentamycin

    Energy Technology Data Exchange (ETDEWEB)

    Mastromarino, A.; Wilson, R.

    1976-11-01

    Oral administration of streptomycin, kanamycin, neomycin, or gentamicin to specific pathogen-free C57 x Af mice in their drinking water (4 mg/ml) for 2 weeks before supralethal whole-body irradiation very significantly prolonged their mean survival times (8.2 to 8.9 days vs 6.9 for controls) to values which exceed those reported for germ-free mice (7.3 days). The total fecal concentrations of aerobes and anaerobes were reduced by kanamycin, neomycin, and gentamicin. Streptomycin reduced the anaerobes significantly, but not the aerobes. Unlike germ-free mice, these antibiotic-treated mice did excrete free bile acids, products of bacterial action. Oral antibiotic treatment was ineffective in altering the transit time of the intestinal mucosal cells. Previously reported studies had indicated a correlation between decreased transit time and increased survival after irradiation. No significant correlation between mean survival time after irradiation and mucosal transit time was observed. The data demonstrate that certain antibiotics alter the character of the intestinal bacterial flora and increase protection against supralethal doses of whole-body irradiation. It is concluded that the mechanisms of radioresistance in antibiotic-treated mice and germ-free mice are different and that in both groups radioresistance is the result of more than elimination of postirradiation infection.

  7. Neomycin Topical

    Science.gov (United States)

    Neomycin, an antibiotic, is used to prevent or treat skin infections caused by bacteria. It is not ... Neomycin comes in cream and ointment that is applied to the skin. Neomycin usually is used one ...

  8. Neomycin inhibits angiogenin-induced angiogenesis.

    Science.gov (United States)

    Hu, G F

    1998-08-18

    A class of angiogenesis inhibitor has emerged from our mechanistic study of the action of angiogenin, a potent angiogenic factor. Neomycin, an aminoglycoside antibiotic, inhibits nuclear translocation of human angiogenin in human endothelial cells, an essential step for angiogenin-induced angiogenesis. The phospholipase C-inhibiting activity of neomycin appears to be involved, because U-73122, another phospholipase C inhibitor, has a similar effect. In contrast, genistein, oxophenylarsine, and staurosporine, inhibitors of tyrosine kinase, phosphotyrosine phosphatase, and protein kinase C, respectively, do not inhibit nuclear translocation of angiogenin. Neomycin inhibits angiogenin-induced proliferation of human endothelial cells in a dose-dependent manner. At 50 microM, neomycin abolishes angiogenin-induced proliferation but does not affect the basal level of proliferation and cell viability. Other aminoglycoside antibiotics, including gentamicin, streptomycin, kanamycin, amikacin, and paromomycin, have no effect on angiogenin-induced cell proliferation. Most importantly, neomycin completely inhibits angiogenin-induced angiogenesis in the chicken chorioallantoic membrane at a dose as low as 20 ng per egg. These results suggest that neomycin and its analogs are a class of agents that may be developed for anti-angiogenin therapy.

  9. Neomycin, Polymyxin, and Bacitracin Topical

    Science.gov (United States)

    Neomycin, polymyxin, and bacitracin combination is used to prevent minor skin injuries such as cuts, scrapes, and burns from becoming infected. Neomycin, polymyxin, and bacitracin are in a class of ...

  10. 2-Deoxystreptamine Conjugates by Truncation–Derivatization of Neomycin

    Directory of Open Access Journals (Sweden)

    Floris L. van Delft

    2010-03-01

    Full Text Available A small library of truncated neomycin-conjugates is prepared by consecutive removal of 2,6-diaminoglucose rings, oxidation-reductive amination of ribose, oxidation-conjugation of aminopyridine/aminoquinoline and finally dimerization. The dimeric conjugates were evaluated for antibacterial activity with a unique hemocyanin-based biosensor. Based on the outcome of these results, a second-generation set of monomeric conjugates was prepared and found to display significant antibacterial activity, in particular with respect to kanamycin-resistant E. coli.

  11. Extremely sensitive sandwich assay of kanamycin using surface-enhanced Raman scattering of 2-mercaptobenzothiazole labeled gold@silver nanoparticles.

    Science.gov (United States)

    Zengin, Adem; Tamer, Ugur; Caykara, Tuncer

    2014-03-19

    Herein, we report the development of extremely sensitive sandwich assay of kanamycin using a combination of anti-kanamycin functionalized hybrid magnetic (Fe3O4) nanoparticles (MNPs) and 2-mercaptobenzothiazole labeled Au-core@Ag-shell nanoparticles as the recognition and surface-enhanced Raman scattering (SERS) substrate, respectively. The hybrid MNPs were first prepared via surface-mediated RAFT polymerization of N-acryloyl-L-glutamic acid in the presence of 2-(butylsulfanylcarbonylthiolsulfanyl) propionic acid-modified MNPs as a RAFT agent and then biofunctionalized with anti-kanamycin, which are both specific for kanamycin and can be collected via a simple magnet. After separating kanamycin from the sample matrix, they were sandwiched with the SERS substrate. According to our experimental results, the limit of detection (LOD) was determined to be 2pg mL(-1), this value being about 3-7 times more than sensitive than the LOD of previously reported results, which can be explained by the higher SERS activity of silver coated gold nanoparticles. The analysis time took less than 10min, including washing and optical detection steps. Furthermore, the sandwich assay was evaluated for investigating the kanamycin specificity on neomycin, gentamycin and streptomycin and detecting kanamycin in artificially contaminated milk.

  12. 21 CFR 558.364 - Neomycin sulfate.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Neomycin sulfate. 558.364 Section 558.364 Food and... in Animal Feeds § 558.364 Neomycin sulfate. (a) Approvals. Type A medicated article: 325 grams per.... (c) (d) Conditions of use. Neomycin sulfate is used as follows: Neomycin Sulfate...

  13. Cardiovascular depressant effects of the neomycin-streptomycin group of antibiotics.

    Science.gov (United States)

    Adams, H R

    1975-01-01

    Cardiovascular depressant effects of the neomycin-streptomycin group of antibiotics (aminoglycoside antibiotics) were examined during pentobarbital anesthesia in cats, dogs, and 4 species of nonhuman primates: owl (Aotus trivirgatus), squirrel (Saimiri sciureus), and rhesus (Macaca mulatta) monkeys, and dog-faced baboons (Papio cynocephalus). Intravenous administration of kanamycin, streptomycin, gentamicin, or neomycin produced various degrees of hypotension and relative bradycardia in all species examined. In surgically prepared (open-chest) baboons, neomycin consistently induced a dose-related depression of myocardial contractile force, maximum dF/dt of myocardial contraction, cardiac output, heart rate, and systolic and diastolic blood pressures. Maximum depression of hemodynamic values usually occurred within 2 to 5 minutes after administration of neomycin; cardiovascular function then gradually returned to control or near control levels within 30 to 60 minutes. Intravenous administration of calcium chloride rapidly reversed the neomycin-mediated alterations of cardiovascular function. Present findings indicated that aminoglycoside antibiotics altered cardiovascular dynamics in anesthetized animals, and indicated that this deleterious action(s) may be related to modification of calcium ion function.

  14. Two-stage method for purification of ceruloplasmin based on its interaction with neomycin.

    Science.gov (United States)

    Sokolov, A V; Kostevich, V A; Romanico, D N; Zakharova, E T; Vasilyev, V B

    2012-06-01

    A two-stage chromatography that yields highly purified ceruloplasmin (CP) from human plasma and from rat and rabbit serum is described. The isolation procedure is based on the interaction of CP with neomycin, and it provides a high yield of CP. Constants of inhibition by gentamycin, kanamycin, and neomycin of oxidase activity of CP in its reaction with p-phenylenediamine were assayed. The lowest K(i) for neomycin (11 µM) corresponded to the highest specific adsorption of CP on neomycin-agarose (10 mg CP/ml of resin). Isolation of CP from 1.4 liters of human plasma using ion-exchange chromatography on UNO-Sphere Q and affinity chromatography on neomycin-agarose yields 348 mg of CP with 412-fold purification degree. Human CP preparation obtained with A(610)/A(280) ~ 0.052 contained neither immunoreactive prothrombin nor active thrombin. Upon storage at 37°C under sterile conditions, the preparation remained stable for two months. Efficient preparation of highly purified CP from rat and rabbit sera treated according to a similar protocol suggests the suitability of our method for isolation of CP from plasma and serum of other animals. The yield of CP in three separate purifications was no less than 78%.

  15. 21 CFR 556.430 - Neomycin.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Neomycin. 556.430 Section 556.430 Food and Drugs... Residues of New Animal Drugs § 556.430 Neomycin. (a) Acceptable daily intake (ADI). The ADI for total residues of neomycin is 6 micrograms per kilogram of body weight per day. (b) Tolerances. Tolerances...

  16. DNase I induced DNA degradation is inhibited by neomycin.

    Science.gov (United States)

    Woegerbauer, M; Burgmann, H; Davies, J; Graninger, W

    2000-03-01

    Preparations of antimicrobials from biotechnological sources containing nucleic acids may serve as vector for the dissemination of resistance genes. An essential prerequisite for the acquisition of a new resistance phenotype in a transformational scenario is the availability of physically intact DNA molecules capable of transforming competent microorganisms. DNA is thought to be an easy target for catabolic processes when present in the natural habitat of bacteria (e.g. gastrointestinal tract, soil) due to the overall presence of nucleolytic enzymes. Aminoglycoside antibiotics are known to display a strong affinity to nucleic acids rendering these compounds to be primary candidates for exerting DNA protective functions in the gastrointestinal tract when applied orally during antibiotic chemotherapy. Using a DNase I protection assay it could be demonstrated that neomycin B at a concentration of 2 mM completely inhibited degradation of plasmid DNA in vitro. No inhibition of degradation was observed with streptomycin and kanamycin and the non-aminoglycoside antibiotics oxytetracycline and ampicillin under identical assay conditions. Thus, neomycin preparations may be able to promote structural integrity of contaminating DNA-fragments in DNase-rich environments.

  17. Determination of neomycin and bacitracin in human or rabbit serum by HPLC-MS/MS.

    Science.gov (United States)

    Mascher, Daniel G; Unger, Christian P; Mascher, Hermann J

    2007-01-17

    The method for the simultaneous determination of neomycin and bacitracin in human or rabbit serum was developed by using ion pairing reversed phase chromatography and tandem mass spectrometry (MS/MS) detection with electrospray (ESI) in positive mode. Both substances elute under these conditions at the same time and also kanamycin as internal standard elutes almost at the same time. The sample preparation was simple-only using 0.1 mL serum by protein precipitation with acetonitrile. Neomycin and bacitracin were detected as two-fold charged ions as well as the internal standard. The calibration range of these quite difficult detectable substances was 0.2-50 microg/mL of serum. The method was validated for both human or rabbit serum. The inter batch precision of quality control samples in human serum for neomycin ranged from 4.46% to 8.99% and for bacitracin from 6.85% to 11.17%. The inter batch accuracy for neomycin ranged from 98.7% to 100.7% and for bacitracin from 99.2% to 103.0%. At lower limit of quantitation (LLOQ) level of 0.2 microg/mL inter batch precision in human serum for neomycin was 12.05% and for bacitracin 11.91%, whereas accuracies were 99.9% for neomycin and 102.7% for bacitracin. Bench top stability in human or rabbit serum was given over three freeze thaw cycles and 4h at room temperature. The method can be considered to be specific and recoveries for sample preparation were high.

  18. HPLC-ELSD determination of kanamycin B in the presence of kanamycin A in fermentation broth.

    Science.gov (United States)

    Zhang, Yong; He, Hui-Min; Zhang, Jin; Liu, Feng-Jiao; Li, Chao; Wang, Bing-Wu; Qiao, Ren-Zhong

    2015-03-01

    A novel method for the direct determination of kanamycin B in the presence of kanamycin A in fermentation broth using high performance liquid chromatography with evaporative light scattering detector (HPLC-ELSD) was developed. An Agilent Technologies C18 column was utilized, evaporation temperature of 40°C and nitrogen pressure of 3.5 bar, the optimized mobile phase was water-acetonitrile (65:35, v/v), containing 11.6 mm heptafluorobutyric acid (isocratic elution with flow rate of 0.5 mL/min) with the gain 11. Kanamycin B was eluted at 5.6 min with an asymmetry factor of 1.827. The method showed good linearity over the concentration range of 0.05 to 0.80 mg/mL for the kanamycin B (r(2) = 0.9987). The intra-day and inter-day coefficients of variation obtained from kanamycin B were less than 4.3%. Mean recovery of kanamycin B from spiked fermentation broth was 95%. The developed method was applied to the determination of kanamycin B without any interference from other constituents in the fermentation broth. This method offers simple, rapid and quantitative detection of kanamycin B. Copyright © 2014 John Wiley & Sons, Ltd.

  19. 21 CFR 522.1484 - Neomycin sulfate sterile solution.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Neomycin sulfate sterile solution. 522.1484... § 522.1484 Neomycin sulfate sterile solution. (a) Specifications. Each milliliter of sterile aqueous solution contains 50 milligrams of neomycin sulfate (equivalent to 35 milligrams of neomycin base).1...

  20. 21 CFR 524.1484a - Neomycin sulfate ophthalmic ointment.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Neomycin sulfate ophthalmic ointment. 524.1484a... § 524.1484a Neomycin sulfate ophthalmic ointment. (a) Specifications. Each gram of the ointment contains 5 milligrams of neomycin sulfate equivalent in activity to 3.5 milligrams of neomycin base....

  1. Neomycin inhibits angiogenin-induced angiogenesis

    OpenAIRE

    1998-01-01

    A class of angiogenesis inhibitor has emerged from our mechanistic study of the action of angiogenin, a potent angiogenic factor. Neomycin, an aminoglycoside antibiotic, inhibits nuclear translocation of human angiogenin in human endothelial cells, an essential step for angiogenin-induced angiogenesis. The phospholipase C-inhibiting activity of neomycin appears to be involved, because U-73122, another phospholipase C inhibitor, has a similar effect. In contrast, genistein, oxophenylarsine, an...

  2. Neomycin inhibits angiogenin-induced angiogenesis

    OpenAIRE

    Hu, Guo-fu

    1998-01-01

    A class of angiogenesis inhibitor has emerged from our mechanistic study of the action of angiogenin, a potent angiogenic factor. Neomycin, an aminoglycoside antibiotic, inhibits nuclear translocation of human angiogenin in human endothelial cells, an essential step for angiogenin-induced angiogenesis. The phospholipase C-inhibiting activity of neomycin appears to be involved, because U-73122, another phospholipase C inhibitor, has a similar effect. In contrast, genistein, oxophenylarsine, an...

  3. Controversy Associated With the Common Component of Most Transgenic Plants – Kanamycin Resistance Marker Gene

    Directory of Open Access Journals (Sweden)

    Srećko Jelenić

    2003-01-01

    Full Text Available Plant genetic engineering is a powerful tool for producing crops resistant to pests, diseases and abiotic stress or crops with improved nutritional value or better quality products. Currently over 70 genetically modified (GM crops have been approved for use in different countries. These cover a wide range of plant species with significant number of different modified traits. However, beside the technology used for their improvement, the common component of most GM crops is the neomycin phosphotransferase II gene (nptII, which confers resistance to the antibiotics kanamycin and neomycin. The nptII gene is present in GM crops as a marker gene to select transformed plant cells during the first steps of the transformation process. The use of antibiotic-resistance genes is subject to controversy and intense debate, because of the likelihood that clinical therapy could be compromised due to inactivation of the oral dose of the antibiotic from consumption of food derived from the transgenic plant, and because of the risk of gene transfer from plants to gut and soil microorganisms or to consumer’s cells. The present article discusses these possibilities in the light of current scientific knowledge.

  4. Stable Agrobacterium-Mediated Transformation of Maritime Pine Based on Kanamycin Selection

    Directory of Open Access Journals (Sweden)

    José M. Alvarez

    2013-01-01

    Full Text Available An efficient transformation protocol based on kanamycin selection was developed for Agrobacterium-mediated transformation of maritime pine embryonal masses. The binary vector pBINUbiGUSint, which contained neomycin phosphotransferase II (nptII as a selectable marker gene and β-glucuronidase (uidA as a reporter gene, was used for transformation studies. Different factors, such as embryogenic line, bacterial strain, bacterial concentration, and coculture duration, were examined and optimized. For selection of transformants, 15 mgL−1 kanamycin was used. The highest transformation efficiency (11.4 events per gram of fresh mass was achieved when a vigorously growing embryonal mass (embryogenic line L01 was cocultivated with Agrobacterium strain AGL1 at the optical density (OD600 nm of 0.3 for 72 h. Evidence of the stable transgene integration was obtained by polymerase chain reaction for the nptII and uidA genes and expression of the uidA gene. Maturation capacity of the transgenic lines was negatively affected by the transformation process. Induction of axillary shoots by preculturing the embryos with benzyladenine allowed overcoming the low maturation rates of some transformed lines. The transgenic embryos were germinated and the axillar shoots were rooted. Transgenic plants were transferred to potting substrate showing normal growth.

  5. Stable Agrobacterium-mediated transformation of maritime pine based on kanamycin selection.

    Science.gov (United States)

    Alvarez, José M; Ordás, Ricardo J

    2013-01-01

    An efficient transformation protocol based on kanamycin selection was developed for Agrobacterium-mediated transformation of maritime pine embryonal masses. The binary vector pBINUbiGUSint, which contained neomycin phosphotransferase II (nptII) as a selectable marker gene and β -glucuronidase (uidA) as a reporter gene, was used for transformation studies. Different factors, such as embryogenic line, bacterial strain, bacterial concentration, and coculture duration, were examined and optimized. For selection of transformants, 15 mgL(-1) kanamycin was used. The highest transformation efficiency (11.4 events per gram of fresh mass) was achieved when a vigorously growing embryonal mass (embryogenic line L01) was cocultivated with Agrobacterium strain AGL1 at the optical density (OD(600 nm)) of 0.3 for 72 h. Evidence of the stable transgene integration was obtained by polymerase chain reaction for the nptII and uidA genes and expression of the uidA gene. Maturation capacity of the transgenic lines was negatively affected by the transformation process. Induction of axillary shoots by preculturing the embryos with benzyladenine allowed overcoming the low maturation rates of some transformed lines. The transgenic embryos were germinated and the axillar shoots were rooted. Transgenic plants were transferred to potting substrate showing normal growth.

  6. The aminoglycoside antibiotic kanamycin damages DNA bases in Escherichia coli: caffeine potentiates the DNA-damaging effects of kanamycin while suppressing cell killing by ciprofloxacin in Escherichia coli and Bacillus anthracis.

    Science.gov (United States)

    Kang, Tina Manzhu; Yuan, Jessica; Nguyen, Angelyn; Becket, Elinne; Yang, Hanjing; Miller, Jeffrey H

    2012-06-01

    The distribution of mutants in the Keio collection of Escherichia coli gene knockout mutants that display increased sensitivity to the aminoglycosides kanamycin and neomycin indicates that damaged bases resulting from antibiotic action can lead to cell death. Strains lacking one of a number of glycosylases (e.g., AlkA, YzaB, Ogt, KsgA) or other specific repair proteins (AlkB, PhrB, SmbC) are more sensitive to these antibiotics. Mutants lacking AlkB display the strongest sensitivity among the glycosylase- or direct lesion removal-deficient strains. This perhaps suggests the involvement of ethenoadenine adducts, resulting from reactive oxygen species and lipid peroxidation, since AlkB removes this lesion. Other sensitivities displayed by mutants lacking UvrA, polymerase V (Pol V), or components of double-strand break repair indicate that kanamycin results in damaged base pairs that need to be removed or replicated past in order to avoid double-strand breaks that saturate the cellular repair capacity. Caffeine enhances the sensitivities of these repair-deficient strains to kanamycin and neomycin. The gene knockout mutants that display increased sensitivity to caffeine (dnaQ, holC, holD, and priA knockout mutants) indicate that caffeine blocks DNA replication, ultimately leading to double-strand breaks that require recombinational repair by functions encoded by recA, recB, and recC, among others. Additionally, caffeine partially protects cells of both Escherichia coli and Bacillus anthracis from killing by the widely used fluoroquinolone antibiotic ciprofloxacin.

  7. 21 CFR 520.1484 - Neomycin.

    Science.gov (United States)

    2010-04-01

    ... containing neomycin or failure to follow withdrawal times may result in illegal drug residues.” (e) Conditions of use—(1) Cattle, swine, sheep, and goats—(i) Amount. 10 mg per pound (/lb) of body weight per.... Labeling shall bear the following warning statements: “A withdrawal period has not been established for...

  8. Mutational biosynthesis of neomycin analogs by a mutant of neomycin-producing Streptomyces fradiae.

    Science.gov (United States)

    Shi, Guanying; Zhang, Xingang; Wu, Lang; Xie, Jin; Tao, Ke; Hou, Taiping

    2011-11-01

    Neomycin, produced by Streptomyces fradiae, has been widely used for the treatment of bacterial infections in clinical and agricultural applications. In this study, a neomycin nonproducing mutant of S. fradiae was obtained by gene disruption technique for mutational biosynthesis. A crucial gene neoC (neo7) which encodes 2-deoxystreptamine (2-DOS) synthases was disrupted. The mutant could resume producing neomycin in the presence of 2-DOS. Salen derivatives of 2-DOS were synthesized and individually added to cultures of the mutant. Antibacterial activity of the mutasynthesis products against Staphylococcus aureus and four plant pathogenic bacteria (Pseudomonas solanacarum, Erwinia carotovora, Xanthomonas oryzae, and Xanthomonas campestris) was detected quantitatively by Oxford cup method. It is suggested that all 2-DOS derivatives were incorporated by the mutant into new active neomycin analogs except for 2-DOS derivative 2d ((1R,2r,3S,4R,6S)-4,6-bis((E)-3,5-di-tert-butyl-2-hydroxybenzylideneamino)cyclohexane-1,2,3-triol). Neomycin analogs produced by feeding 2-DOS derivative 2a ((1R,2r,3S,4R,6S)-4,6-bis((E)-2 hydroxybenzylideneamino)cyclohexane-1,2,3-triol) to cultures of the mutant displayed a similar antibacterial activity with neomycin produced by wild strain.

  9. 21 CFR 524.1200b - Kanamycin ophthalmic aqueous solution.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Kanamycin ophthalmic aqueous solution. 524.1200b Section 524.1200b Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES... § 524.1200b Kanamycin ophthalmic aqueous solution. (a) Specifications. The drug, which is in an...

  10. Immunoassay analysis of Kanamycin in serum using the Tobramycin kit

    NARCIS (Netherlands)

    Dijkstra, J A; Voerman, A J; Greijdanus, B; Touw, D J; Alffenaar, J W C

    2016-01-01

    Kanamycin is one of the aminoglycosides used in the treatment of multidrug resistant tuberculosis. Blood concentrations of kanamycin are predictive for the treatment efficacy and the occurrence of side effects and dose adjustments can be needed to optimize therapy. However, an immunoassay method for

  11. Immunoassay Analysis of Kanamycin in Serum Using the Tobramycin Kit

    NARCIS (Netherlands)

    Dijkstra, J.A.; Voerman, A. J.; Greijdanus, B.; Touw, D. J.; Alffenaar, J. W. C.

    2016-01-01

    Kanamycin is one of the aminoglycosides used in the treatment of multidrug-resistant tuberculosis. Blood concentrations of kanamycin are predictive for the treatment efficacy and the occurrence of side effects, and dose adjustments can be needed to optimize therapy. However, an immunoassay method fo

  12. Recognition of HIV TAR RNA by triazole linked neomycin dimers.

    Science.gov (United States)

    Kumar, Sunil; Arya, Dev P

    2011-08-15

    A series of neomycin dimers have been synthesized using 'click chemistry' with varying linker functionality and length to target the TAR RNA region of HIV virus. TAR (trans activation response) RNA region, a 59 base pair stem loop structure located at 5'-end of all nascent HIV-1 transcripts interacts with a key regulatory protein, Tat, and necessitates the replication of HIV-1 virus. Neomycin, an aminosugar, has been shown to exhibit more than one binding site with HIV TAR RNA. Multiple TAR binding sites of neomycin prompted us to design and synthesize a small library of neomycin dimers using click chemistry. The binding between neomycin dimers and HIV TAR RNA was characterized using spectroscopic techniques including FID (Fluorescent Intercalator Displacement) titration and UV-thermal denaturation. UV thermal denaturation studies demonstrate that neomycin dimer binding increase the melting temperature (T(m)) of the HIV TAR RNA up to 10°C. Ethidium bromide displacement titrations revealed nanomolar IC(50) between neomycin dimers and HIV TAR RNA, whereas with neomycin, a much higher IC(50) in the micromolar range is observed.

  13. Screening Method for Transgenic Maize with Kanamycin Resistance

    Institute of Scientific and Technical Information of China (English)

    LU Meng-yu; DONG Fu-shuang; ZHANG Jun-min; WANG Hai-bo

    2011-01-01

    [Objective] The paper was to establish simple and effective method to screen marker gene in maize with kanamycin resistance.[Method] Using inbred line "Chang 7-2" and hybrid "Zhengdan 958" of maize as test materials,their seeds were soaked with different concentrations and volumes of kanamycin for 3 and 4 d,respectively,the rate of albino seedlings and average seedling height after sowing for 10 d were investigated.[Result] The rate of albino seedlings not only was related to kanamycin concentration,but also had relationship with solution volume during soaking process.The difference between inbred line and hybrid was no significant.When 100 ml of kanamycin solution with concentration of 200 mg/L was used to soak seeds for 3 d,the rate of albino seedlings basically could reach 100%.When 100 ml of kanamycin solution with concentration of 100 mg/L was used to soak 20 seeds for 3 d to carry out resistance screening,the accuracy was up to 84.8% after verifying the screening test of T1 transgenic maize plants.[Conclusion] The method was feasible,which could be used as a simple method for screening transgenic gene maize with kanamycin resistance.

  14. Evidence that neomycin inhibits human cytomegalovirus infection of fibroblasts.

    Science.gov (United States)

    Lobert, P E; Hober, D; Delannoy, A S; Wattré, P

    1996-01-01

    The effect of phosphoinositide-binding aminoglycosides, such as neomycin, gentamicin and streptomycin, on human cytomegalovirus (HCMV) infection of human fibroblasts MRC-5 was studied. The inhibition of HCMV infection was obtained with all of these molecules but neomycin was more effective than the others. We showed that the inoculation of the cells with cell-free viral suspension in presence of neomycin concentrations above 5 mM at 37 degrees C, inhibited more than 98% the HCMV infection. However, the preincubation of the fibroblasts with neomycin at 4 degrees C, before the removal of the drug and the inoculation of the cells, induced only a 30% decrease in the number of infected cells. Addition of neomycin after the HCMV-binding at 4 degrees C or the infection of the cells was less efficient to inhibit HCMV infection than the standard incubation of neomycin during inoculation of the fibroblasts. Indeed, 1 hour after the inoculation of the cells at 37 degrees C, neomycin still inhibited HCMV infection, but 4 hours after the inoculation, this drug had no effect on HCMV infection. Our findings demonstrated that neomycin must be present at the time of infection in order to exert a full inhibiting effect. The effect of neomycin on the HCMV infection was almost immediate upon the addition of the drug (binding and/or internalization) and after the virus internalization (inhibition of immediate-early events). We suggest that neomycin and other aminoglycoside antibiotics may interact with HCMV glycoproteins for binding to similar structural features of cell surface heparan sulfate proteoglycans and may inhibit HCMV infection in fibroblasts by disrupting phosphoinositide-mediated events in the cells.

  15. Development of Enzyme-linked Immunosorbent Assay for Neomycin%新霉素ELISA检测方法的建立

    Institute of Scientific and Technical Information of China (English)

    刘沙洲; 桑小雪; 欧阳华学; 雷绍荣; 白林含

    2011-01-01

    In this study, we describe the advantages and disadvantages of direct competitive enzyme-linked immunosorbent assay (dc-ELISA) and indirect competitive ELISA (idc-ELISA) and ELISA methods for the detection of neomycin. Antineomycin polyclonal antibodies were prepared and used to detect neomycin by dc-ELISA and idc-ELISA. The cross-reaction rates of prepared anti-neomycin polyclonal antibodies with gentamincin and kanamycin were 2.04% and 0.02%, respectively, and with ampicillin, erythromycin and tetracycline all less than 0.01%. The accuracy and recovery of idc-ELISA were tested with an intra-plate error of less than 4%, an inter-plate error of less than 11% and a recovery between 135.5% and 191.3%. The detection limits of dc-ELISA and idc-ELISA were 28.58 ng/mL and 51.74 ng/mL, respectively, both of which were below the national maximum residue limit (MRL) of 500 pg/kg. Therefore, a dc-ELISA method and an idc-ELISA method to detect neomycin have successfully established. Further, the idc-ELISA method where the working conditions were better optimized can be used for the development of neomycin test kit.%目的:比较直接和间接竞争酶联免疫法(enzyme linked immunosorbent assay,ELISA)的优缺点,建立新霉素残留ELISA检测方法。方法:利用自制的新霉素多克隆抗体,采用直接竞争和间接竞争ELISA方法检测新霉素残留,并比较两种方法的优缺点。结果:新霉素抗血清和庆大霉素的交叉反应率为2.04%,和卡那霉素的交叉反应率为0.02%,和氨苄青霉素、红霉素、四环素的交叉反应率均小于0.01%。初步测试新霉素间接竞争ELISA法的准确性和回收率。板内误差小于4%,板间误差小于11%,回

  16. Suitability of a liquid chromatography assay of neomycin sulfate to replace the microbiological assay for neomycin in USP Monographs.

    Science.gov (United States)

    Hanko, Valoran P; Rohrer, Jeffrey S

    2010-01-01

    The current USP National Formulary contains 65 Monographs for drug formulations containing neomycin. All 65 Monographs prescribe a bioassay for neomycin assay. This bioassay, based on cell culture, is labor intensive, has poor precision, and cannot be adapted for purity or identification. High-performance anion-exchange chromatography with integrated pulsed amperometric detection (HPAE-IPAD), a liquid chromatography technique, has been shown to be suitable for neomycin purity analysis and neomycin assay of an over-the-counter first aid cream (Hanko and Rohrer [17]). Here we propose that an HPAE-IPAD assay can replace the bioassay in the 65 neomycin-containing Monographs. We applied the HPAE-IPAD assay to four neomycin-containing drug products representing the four classes of formulations found in the 65 Monographs, liquid, solid, suspension, and cream. Each drug was analyzed with two chromatography systems, and on 3 separate days. For all products, HPAE-IPAD measurements were precise and accurate with respect to the label concentrations. There was also high accuracy for spike recovery of neomycin from the four drug products throughout 70-150% of the labeled concentration. These results suggest that an HPAE-IPAD assay would be an accurate assay for neomycin, and would be faster and more precise than the current bioassay.

  17. 21 CFR 524.1484i - Neomycin sulfate, hydrocortisone acetate, sterile ointment.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Neomycin sulfate, hydrocortisone acetate, sterile... NEW ANIMAL DRUGS § 524.1484i Neomycin sulfate, hydrocortisone acetate, sterile ointment. (a) Specifications. The drug contains 5 milligrams of neomycin sulfate, equivalent to 3.5 milligrams of neomycin...

  18. Probing the recognition surface of a DNA triplex: Binding studies with intercalator-neomycin conjugates

    OpenAIRE

    XUE Liang; Xi, Hongjuan; Kumar, Sunil; Gray, David; Davis, Erik; Hamilton, Paris; Skirba, Michael; Arya, Dev P.

    2010-01-01

    Thermodynamic studies on the interactions between intercalator-neomycin conjugates and a DNA polynucleotide triplex [poly(dA)•2poly(dT)] were conducted. To draw a complete picture of such interactions, naphthalenedimide-neomycin (3) and anthraquinone-neomycin (4) were synthesized and used together with two other analogues, previously synthesized pyrene-neomycin (1) and BQQ-neomycin (2), in our investigations. A combination of experiments including UV denaturation, circular dichroism (CD) titr...

  19. Neomycin is more efficient than streptomycin in suppressing frameshift mutations.

    Science.gov (United States)

    Phoenix, P; Gravel, M; Herrington, M B; Brakier-Gingras, L

    1985-12-01

    The effects of streptomycin and neomycin on the phenotypic suppression of frameshift mutations in the lacZ gene of Escherichia coli and on the efficiency of suppression of amber mutations in T4 phage by the informational supE tRNA nonsense suppressor were compared. Neomycin stimulated much more efficiently than streptomycin the phenotypic suppression of frameshift mutations. Because neomycin favors mismatches of the central codon base whereas streptomycin favors mismatches of the first codon base, this result suggests that mismatching of the central codon base pair and shifting of the reading frame are two correlated phenomena. In contrast, both streptomycin and neomycin stimulated about equally the efficiency of the tRNA nonsense suppressor, an effect probably related to their interference with the proofreading control in tRNA selection.

  20. Label-free gold nanoparticles for the determination of neomycin

    Science.gov (United States)

    Apyari, Vladimir V.; Dmitrienko, Stanislava G.; Arkhipova, Viktoriya V.; Atnagulov, Aydar G.; Gorbunova, Mariya V.; Zolotov, Yury A.

    2013-11-01

    A new spectrophotometric method for the determination of neomycin has been developed. The method is based on aggregation of label-free gold nanoparticles leading to change in absorption spectra and color of the solution. Influence of different factors (the concentration of ethylenediaminetetraacetate (EDTA), pH, the concentrations of neomycin and the nanoparticles) on the aggregation and analytical performance of the method was investigated. EDTA plays an important role not only as a masking agent to eliminate interferences of metal cations but strongly affects the sensitivity of the nanoparticles relative to neomycin. The method allows to determine neomycin with detection limit of 28 ng mL-1. It was applied to analysis of eye- and ear-drops. The sample pretreatment is simply done by diluting the formulation with water.

  1. Kanamycin resistance during in vitro development of pollen from transgenic tomato plants

    NARCIS (Netherlands)

    Bino, R.J.; Hille, J.; Franken, J.

    1987-01-01

    Effects of kanamycin on pollen germination and tube growth of pollen from non-transformed plants and from transgenic tomato plants containing a chimaeric kanamycin resistance gene were determined. Germination of pollen was not affected by the addition of kanamycin to the medium in both genotypes. Ka

  2. Screening for engineered neomycin riboswitches that control translation initiation.

    Science.gov (United States)

    Weigand, Julia E; Sanchez, Martin; Gunnesch, Ewald-Bernd; Zeiher, Sabrina; Schroeder, Renee; Suess, Beatrix

    2008-01-01

    Riboswitches are genetic control elements that regulate gene expression in a small molecule-dependent way. We developed a two-stage strategy of in vitro selection followed by a genetic screen and identified several artificial small molecule-binding riboswitches that respond to the aminoglycoside neomycin. Structure-function relationships and structural probing revealed that they adopt the general neomycin-binding motif. They display no sequence similarities to in vitro selected neomycin aptamers but contain parts of the decoding site that is the binding site for neomycin on the ribosomal RNA. We propose a model of a composed binding pocket of an internal loop as primary docking site and a terminal flaplike loop structure fixing neomycin in a sandwich-like manner. Such binding pockets characterized by multiple contacts between ligand and RNA are described for both natural and engineered riboswitches. We anticipate that combination of in vitro selection and in vivo screening is a useful strategy to identify RNA molecules with a desired functionality.

  3. 21 CFR 524.1600a - Nystatin, neomycin, thiostrepton, and triamcinolone acetonide ointment.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Nystatin, neomycin, thiostrepton, and... TOPICAL DOSAGE FORM NEW ANIMAL DRUGS § 524.1600a Nystatin, neomycin, thiostrepton, and triamcinolone... base ointment contains: 100,000 units of nystatin; neomycin sulfate equivalent to 2.5 milligrams...

  4. Possible role for TRPV1 in neomycin-induced inhibition of visceral hypersensitivity in rat

    NARCIS (Netherlands)

    R.M. van den Wijngaard; O. Welting; D.C. Bulmer; M.M. Wouters; K. Lee; W.J. de Jonge; G.E. Boeckxstaens

    2009-01-01

    Transient receptor ion channel 1 (TRPV1) is a nociceptor involved in visceral hypersensitivity. Aminoglycosides like neomycin are not only potent antibiotics but in vitro data suggest that neomycin also acts as a TRPV1-antagonist and alleviates somatic pain responses. To what extent neomycin reduces

  5. 21 CFR 524.1883 - Prednisolone sodium phosphate-neomycin sulfate ophthalmic ointment.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Prednisolone sodium phosphate-neomycin sulfate... DOSAGE FORM NEW ANIMAL DRUGS § 524.1883 Prednisolone sodium phosphate-neomycin sulfate ophthalmic ointment. (a) Specifications. Prednisolone sodium phosphate-neomycin sulfate ophthalmic ointment...

  6. 21 CFR 524.1880 - Prednisolone-neomycin sulfate ophthalmic ointment.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Prednisolone-neomycin sulfate ophthalmic ointment... ANIMAL DRUGS § 524.1880 Prednisolone-neomycin sulfate ophthalmic ointment. (a) Specifications. Prednisolone-neomycin sulfate ophthalmic ointment contains 2 milligrams prednisolone and 5 milligrams...

  7. 40 CFR 174.521 - Neomycin phosphotransferase II; exemption from the requirement of a tolerance.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Neomycin phosphotransferase II...-INCORPORATED PROTECTANTS Tolerances and Tolerance Exemptions § 174.521 Neomycin phosphotransferase II; exemption from the requirement of a tolerance. Residues of the neomycin phosphotransferase II (NPTII)...

  8. Ecabet sodium alleviates neomycin-induced hair cell damage.

    Science.gov (United States)

    Rah, Yoon Chan; Choi, June; Yoo, Myung Hoon; Yum, Gunhwee; Park, Saemi; Oh, Kyoung Ho; Lee, Seung Hoon; Kwon, Soon Young; Cho, Seung Hyun; Kim, Suhyun; Park, Hae-Chul

    2015-12-01

    Ecabet sodium (ES) is currently applied to some clinical gastrointestinal disease primarily by the inhibition of the ROS production. In this study, the protective role of ES was evaluated against the neomycin-induced hair cell loss using zebrafish experimental animal model. Zebrafish larvae (5-7 dpf), were treated with each of the following concentrations of ES: 5, 10, 20, 40, and 80 μg/mL for 1 h, followed by 125 μM neomycin for 1h. The positive control group was established by 125 μM neomycin-only treatment (1h) and the negative control group with no additional chemicals was also established. Hair cells inside four neuromasts ( SO1, SO2, O1, OC1) were assessed using fluorescence microscopy (n = 10). Hair cell survival was calculated as the mean number of viable hair cells for each group. Apoptosis and mitochondrial damage were investigated using special staining (TUNEL and DASPEI assay, respectively), and compared among groups. Ultrastructural changes were evaluated using scanning electron microscopy. Pre-treatment group with ES increased the mean number of viable hair cells as a dose-dependent manner achieving almost same number of viable hair cells with 40 μM/ml ES treatment (12.98 ± 2.59 cells) comparing to that of the negative control group (14.15 ± 1.39 cells, p = 0.72) and significantly more number of viable hair cells than that of the positive control group (7.45 ± 0.91 cells, p neomycin treatment than the negative control group and significantly decreased down to 105% with the pre-treatment with 40 μM/ml ES (n = 40, p = 0.04). A significantly less number of TUNEL-positive cells (reflecting apoptosis, p neomycin-induced hair cell loss possibly by reducing apoptosis, mitochondrial damages, and the ROS generation.

  9. Scar formation in mice deafened with kanamycin and furosemide.

    Science.gov (United States)

    Żak, Magdalena; van der Linden, Cynthia A; Bezdjian, Aren; Hendriksen, Ferry G; Klis, Sjaak F L; Grolman, Wilko

    2016-08-01

    In mammals, hair cell loss is irreversible and leads to hearing loss. To develop and test the functioning of different strategies aiming at hair cell regeneration, animal models of sensorineural hearing loss are essential. Although cochleae of these animals should lack hair cells, supporting cells should be preserved forming an environment for the regenerated hair cells. In this study, we investigated how ototoxic treatment with kanamycin and furosemide changes the structure of cochlear sensory epithelium in mice. The study also compared different tissue preparation protocols for scanning electron microscopy (SEM). Cochleae were collected from deafened and nondeafened mice and further processed for plastic mid modiolar sections and SEM. For comparing SEM protocols, cochleae from nondeafened mice were processed using three protocols: osmium-thiocarbohydrazide-osmium (OTO), tannic acid-arginine-osmium, and the conventional method with gold-coating. The OTO method demonstrated optimal cochlear tissue preservation. Histological investigation of cochleae of deafened mice revealed that the supporting cells enlarged and ultimately replaced the lost hair cells forming types 1 and 2 phalangeal scars in a base towards apex gradient. The type 3 epithelial scar, flattened epithelium, has not been seen in analysed cochleae. The study concluded that mice deafened with kanamycin and furosemide formed scars containing supporting cells, which renders this mouse model suitable for testing various hair cell regeneration approaches. Microsc. Res. Tech. 79:766-772, 2016. © 2016 Wiley Periodicals, Inc.

  10. Synthesis and combinational antibacterial study of 5''-modified neomycin.

    Science.gov (United States)

    Zhang, Jianjun; Keller, Katherine; Takemoto, Jon Y; Bensaci, Mekki; Litke, Anthony; Czyryca, Przemyslaw Greg; Chang, Cheng-Wei Tom

    2009-10-01

    A library of 5''-modified neomycin derivatives were synthesized for an antibacterial structure-activity optimization strategy. Two leads exhibited prominent activity against both methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE). Antibacterial activities were measured when combined with other clinically used antibiotics. Significant synergistic activities were observed, which may lead to the development of novel therapeutic practices in the battle against infectious bacteria.

  11. Minocycline protection of neomycin induced hearing loss in gerbils.

    Science.gov (United States)

    Robinson, Alan M; Vujanovic, Irena; Richter, Claus-Peter

    2015-01-01

    This animal study was designed to determine if minocycline ameliorates cochlear damage is caused by intratympanic injection of the ototoxic aminoglycoside antibiotic neomycin. Baseline auditory-evoked brainstem responses were measured in gerbils that received 40 mM intratympanic neomycin either with 0, 1.2, or 1.5 mg/kg intraperitoneal minocycline. Four weeks later auditory-evoked brainstem responses were measured and compared to the baseline measurements. Minocycline treatments of 1.2 mg/kg and 1.5 mg/kg resulted in significantly lower threshold increases compared to 0 mg/kg, indicating protection of hearing loss between 6 kHz and 19 kHz. Cochleae were processed for histology and sectioned to allow quantification of the spiral ganglion neurons and histological evaluation of organ of Corti. Significant reduction of spiral ganglion neuron density was demonstrated in animals that did not receive minocycline, indicating that those receiving minocycline demonstrated enhanced survival of spiral ganglion neurons, enhanced survival of sensory hairs cells and spiral ganglion neurons, and reduced hearing threshold elevation correlates with minocycline treatment demonstrating that neomycin induced hearing loss can be reduced by the simultaneous application of minocycline.

  12. Analysis of the Kanamycin in Raw Milk Using the Suspension Array

    Directory of Open Access Journals (Sweden)

    Yanfei Wang

    2013-01-01

    Full Text Available With the monoclonal antibody against kanamycin being prepared successfully, a bead-based indirect competitive fluorescent immunoassay was developed to detect kanamycin in milk. The fact that there was no significant cross-reaction with other aminoglycoside antibiotics implied that the monoclonal antibody was highly specific for kanamycin. The limit of detection (LOD and the 50% inhibition concentration (IC50 in raw milk were 3.2 ng/mL and 52.5 ng/mL, respectively. Using the method developed in this study, the kanamycin concentrations were monitored in raw milk after the intramuscular administration of kanamycin in sick cows. Compared to the conventional enzyme-linked immunosorbent assay (ELISA, the method using the suspension array system was more sensitive. The results obtained in the present study showed a good correlation with that of the ELISA.

  13. Possible role for TRPV1 in neomycin-induced inhibition of visceral hypersensitivity in rat.

    Science.gov (United States)

    van den Wijngaard, R M; Welting, O; Bulmer, D C; Wouters, M M; Lee, K; de Jonge, W J; Boeckxstaens, G E

    2009-08-01

    Transient receptor ion channel 1 (TRPV1) is a nociceptor involved in visceral hypersensitivity. Aminoglycosides like neomycin are not only potent antibiotics but in vitro data suggest that neomycin also acts as a TRPV1-antagonist and alleviates somatic pain responses. To what extent neomycin reduces visceral hypersensitivity remains unknown. Therefore, we aimed to investigate whether neomycin can inhibit in vivo TRPV1-dependent hypersensitivity responses in two rat models of visceral pain. In the first model rats were pretreated with intraperitoneal (i.p.) capsazepine, the selective TRPV1 antagonist SB-705498, neomycin or vehicle alone and 30 min later instilled with intracolonic TRPV1-activating capsaicin. Likewise, rats were pretreated with 10 days oral neomycin and then subjected to intracolonic capsaicin. The visceromotor response (VMR) to distension was measured before and after capsaicin application. In addition, the VMR to distension was measured in adult maternal separated rats before and after acute stress. Before the 2nd distension protocol these rats were treated with i.p. neomycin, amoxycillin or vehicle alone. Our results showed that capsaicin administration induced an enhanced VMR to distension that was prevented by i.p. capsazepine, SB-705498 and neomycin. Oral neomycin treatment changed bacterial faecal content but could not inhibit capsaicin induced visceral hypersensitivity. In maternal separated rats acute stress induced an enhanced response to distension that was reversed by i.p. neomycin, but not amoxycillin. These data indicate that (i.p.) neomycin can inhibit visceral hypersensitivity to distension in a nonbactericidal manner and suggest that TRPV1-modulation may be involved.

  14. Surveillance of Kanamycin Resistance to Escherichia coli from Swine by Digoxigenin-labled Plasmid Probe

    Institute of Scientific and Technical Information of China (English)

    YANG Han-chun; ZHAO Jing; LIU Jin-hua; ZHA Zhen-lin; CHEN Yan-hong

    2002-01-01

    A 4.34kb EcoR I fragment of kanamycin resistance plasmid from pET - 9a was purified by a DNA purification kit. The fragment was labeled with digoxigenin-dUTP with a commercial kit. A dot-blot hybridization and a colony hybridization test with the probe were successfully developed for the surveillance of Kanamycin resistance to E. coli from swine. It was shown that the methods obtained 100% concordance in a positive tate. It was indicated that the method was available for the surveillance of kanamycin resistance to E.coli from swine.

  15. 21 CFR 524.1484g - Neomycin sulfate-thiabendazole-dexamethasone solution.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Neomycin sulfate-thiabendazole-dexamethasone solution. 524.1484g Section 524.1484g Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND... NEW ANIMAL DRUGS § 524.1484g Neomycin sulfate-thiabendazole-dexamethasone solution. (a)...

  16. 21 CFR 524.1484e - Neomycin sulfate and polymyxin B sulfate ophthalmic solution.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Neomycin sulfate and polymyxin B sulfate ophthalmic solution. 524.1484e Section 524.1484e Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF... DOSAGE FORM NEW ANIMAL DRUGS § 524.1484e Neomycin sulfate and polymyxin B sulfate ophthalmic solution....

  17. 21 CFR 524.981c - Fluocinolone acetonide, neomycin sulfate cream.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Fluocinolone acetonide, neomycin sulfate cream. 524.981c Section 524.981c Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN... ANIMAL DRUGS § 524.981c Fluocinolone acetonide, neomycin sulfate cream. (a) Specifications. The...

  18. 21 CFR 524.1484k - Neomycin sulfate, prednisolone, tetracaine, and squalane topical-otic suspension.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Neomycin sulfate, prednisolone, tetracaine, and... TOPICAL DOSAGE FORM NEW ANIMAL DRUGS § 524.1484k Neomycin sulfate, prednisolone, tetracaine, and squalane topical-otic suspension. (a) Specifications. Each milliliter of suspension contains 5 milligrams...

  19. From GUIDON to NEOMYCIN and HERACLES in Twenty Short Lessons. Final Report. ONR Technical Report #20.

    Science.gov (United States)

    Clancey, William J.

    This paper reviews the research leading from the GUIDON rule-based tutoring system, including the reconfiguration of MYCIN into NEOMYCIN and NEOMYCIN's generalization into the heuristic classification shell, HERACLES. The presentation is organized chronologically around pictures and dialogues that represent turning points and crystallize the basic…

  20. 21 CFR 524.1484 - Neomycin sulfate ophthalmic and topical dosage forms.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Neomycin sulfate ophthalmic and topical dosage forms. 524.1484 Section 524.1484 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND... NEW ANIMAL DRUGS § 524.1484 Neomycin sulfate ophthalmic and topical dosage forms....

  1. Probing the recognition surface of a DNA triplex: binding studies with intercalator-neomycin conjugates.

    Science.gov (United States)

    Xue, Liang; Xi, Hongjuan; Kumar, Sunil; Gray, David; Davis, Erik; Hamilton, Paris; Skriba, Michael; Arya, Dev P

    2010-07-06

    Thermodynamic studies on the interactions between intercalator-neomycin conjugates and a DNA polynucleotide triplex [poly(dA).2poly(dT)] were conducted. To draw a complete picture of such interactions, naphthalene diimide-neomycin (3) and anthraquinone-neomycin (4) conjugates were synthesized and used together with two other analogues, previously synthesized pyrene-neomycin (1) and BQQ-neomycin (2) conjugates, in our investigations. A combination of experiments, including UV denaturation, circular dichroism (CD) titration, differential scanning calorimetry (DSC), and isothermal titration calorimetry (ITC), revealed that all four conjugates (1-4) stabilized poly(dA).2poly(dT) much more than its parent compound, neomycin. UV melting experiments clearly showed that the temperature (T(m3-->2)) at which poly(dA).2poly(dT) dissociated into poly(dA).poly(dT) and poly(dT) increased dramatically (>12 degrees C) in the presence of intercalator-neomycin conjugates (1-4) even at a very low concentration (2 muM). In contrast to intercalator-neomycin conjugates, the increment of T(m3-->2) of poly(dA).2poly(dT) induced by neomycin was negligible under the same conditions. The binding preference of intercalator-neomycin conjugates (1-4) to poly(dA).2poly(dT) was also confirmed by competition dialysis and a fluorescent intercalator displacement assay. Circular dichroism titration studies revealed that compounds 1-4 had slightly larger binding site size ( approximately 7-7.5) with poly(dA).2poly(dT) as compared to neomycin ( approximately 6.5). The thermodynamic parameters of these intercalator-neomycin conjugates with poly(dA).2poly(dT) were derived from an integrated van't Hoff equation using the T(m3-->2) values, the binding site size numbers, and other parameters obtained from DSC and ITC. The binding affinity of all tested ligands with poly(dA).2poly(dT) increased in the following order: neomycin intercalator-neomycin conjugates for poly(dA).2poly(dT) increases as a function of

  2. Comparison of the misreading induced by streptomycin and neomycin.

    Science.gov (United States)

    Grisé-Miron, L; Noreau, J; Melançon, P; Brakier-Gingras, L

    1981-11-27

    In a poly(U)-programmed translation system, neomycin stimulates the misincorporation of tyrosine and of serine which, according to Thompson and Stone (Thompson, R.C. and Stone, P.J. (1977) Proc. Natl. Acad. Sci. USA. 74, 198-202), are normally rejected at an initial discrimination step during the binding of charged tRNAs to the ribosome. In contrast, streptomycin favors the misincorporation of isoleucine which is normally rejected at a subsequent GTP-dependent discrimination step, the so-called proofreading step. The labeling of the ribosome with N-ethylmaleimide mimics the effect of streptomycin in that it stimulates the misincorporation of isoleucine but not of tyrosine or serine. This effect is correlated with the labeling of protein S18 but not with that of protein S1. These observations indicate that the sulfhydryl group of protein S18 is located within a ribosomal domain involved in the proofreading control of tRNA selection. Taking into account our previous results that streptomycin and neomycin perturb ribosomal areas around the sulfhydryl groups of proteins S18 and S1, respectively, we suggest that these antibiotics induce misreading by different mechanisms which are linked to such perturbations.

  3. 21 CFR 524.1484b - Neomycin sulfate, isoflupredone acetate, tetracaine hydrochloride, and myristyl-gamma-picolinium...

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Neomycin sulfate, isoflupredone acetate... Neomycin sulfate, isoflupredone acetate, tetracaine hydrochloride, and myristyl-gamma-picolinium chloride, topical powder. (a) Specifications. The product contains 5 milligrams of neomycin sulfate, equivalent to...

  4. 21 CFR 524.154 - Bacitracin or bacitracin zinc-neomycin sulfate-polymyxin B sulfate ophthalmic ointment.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Bacitracin or bacitracin zinc-neomycin sulfate... TOPICAL DOSAGE FORM NEW ANIMAL DRUGS § 524.154 Bacitracin or bacitracin zinc-neomycin sulfate-polymyxin B...: (1) To 000009; each gram contains 500 units of bacitracin, 3.5 milligrams of neomycin, and...

  5. Overexpression of the Chromosomally Encoded Aminoglycoside Acetyltransferase eis Confers Kanamycin Resistance in Mycobacterium tuberculosis

    National Research Council Canada - National Science Library

    M. Analise Zaunbrecher; R. David Sikes; Beverly Metchock; Thomas M. Shinnick; James E. Posey

    2009-01-01

    .... The aminoglycosides kanamycin and amikacin are important bactericidal drugs used to treat MDR TB, and resistance to one or both of these drugs is a defining characteristic of extensively drug-resistant TB...

  6. Impact of kanamycin on melanogenesis and antioxidant enzymes activity in melanocytes--an in vitro study.

    Science.gov (United States)

    Wrześniok, Dorota; Otręba, Michał; Beberok, Artur; Buszman, Ewa

    2013-12-01

    Aminoglycosides, broad spectrum aminoglycoside antibiotics, are used in various infections therapy due to their good antimicrobial characteristics. However, their adverse effects such as nephrotoxicity and auditory ototoxicity, as well as some toxic effects directed to pigmented tissues, complicate the use of these agents. This study was undertaken to investigate the effect of aminoglycoside antibiotic-kanamycin on viability, melanogenesis and antioxidant enzymes activity in cultured human normal melanocytes (HEMa-LP). It has been demonstrated that kanamycin induces concentration-dependent loss in melanocytes viability. The value of EC50 was found to be ~6.0 mM. Kanamycin suppressed melanin biosynthesis: antibiotic was shown to inhibit cellular tyrosinase activity and to reduce melanin content in normal human melanocytes. Significant changes in the cellular antioxidant enzymes: SOD, CAT and GPx were stated in melanocytes exposed to kanamycin. Moreover, it was observed that kanamycin caused depletion of antioxidant defense sytem. It is concluded that the inhibitory effect of kanamycin on melanogenesis and not sufficient antioxidant defense mechanism in melanocytes in vitro may explain the potential mechanisms of undesirable side effects of this drug directed to pigmented tissues in vivo. © 2013 Wiley Periodicals, Inc.

  7. Polyethylene oxide (PEO)-hyaluronic acid (HA) nanofibers with kanamycin inhibits the growth of Listeria monocytogenes.

    Science.gov (United States)

    Ahire, J J; Robertson, D D; van Reenen, A J; Dicks, L M T

    2017-02-01

    Listeria monocytogenes is well known to cause prosthetic joint infections in immunocompromised patients. In this study, polyethylene oxide (PEO) nanofibers, containing kanamycin and hyaluronic acid (HA), were prepared by electrospinning at a constant electric field of 10kV. PEO nanofibers spun with 0.2% (w/v) HA and 1% (w/v) kanamycin had a smooth, bead-free structure at 30-35% relative humidity. The average diameter of the nanofibers was 83±20nm. Attenuated total reflectance (ATR)-Fourier transform infrared (FTIR) spectroscopy indicated that kanamycin was successfully incorporated into PEO/HA matrix. The presence of kanamycin affects the thermal properties of PEO/HA nanofibers, as shown by differential scanning calorimetry (DSC) and thermogravimetric analyses (TGA). The kanamycin-PEO-HA nanofibers (1mg; 47±3μg kanamycin) inhibited the growth of L. monocytogenes EDGe by 62%, as compared with PEO-HA nanofibers, suggesting that it may be used to coat prosthetic implants to prevent secondary infections.

  8. Determination of neomycin in the form of neomycin derivative with dabsyl chloride by thin layer chromatography and densitometry.

    Science.gov (United States)

    Hubicka, Urszula; Zuromska-Witek, Barbara; Piotrowska, Joanna; Krzek, Jan

    2015-01-01

    A thin layer chromatographic-densitometric method has been developed for identification and quantitative determination of neomycin derivative with dabsyl chloride. The analysis of antibiotic was achieved on the silica gel TLC plates with fluorescent indicator with n-butanol--2-butanone--25% ammonia--water (10 : 6 : 2 : 2, v/v/v/v) as the mobile phase. The densitometric measurements were made at 460 nm. Under these conditions good separation of chosen aminoglycoside antibiotic from reagent used to make a complex was obtained. The method is characterized by high sensitivity, LOD from 0.1953 μg per band and LOQ from 0.5918 μg per band, wide linearity range from 0.5918 to 2.1960 μg per band for neomycin. The precision of the method was good; RSD varied from 1.17 to 2.05%. Satisfactory results of validation of the method were also confirmed by determination of selected antibiotic in pharmaceutical commercial preparation. The results obtained by TLC-densitometric method were compared with those obtained by spectrophotometric method.

  9. Ultra-sensitive detection of kanamycin for food safety using a reduced graphene oxide-based fluorescent aptasensor

    Science.gov (United States)

    Ha, Na-Reum; Jung, In-Pil; La, Im-Joung; Jung, Ho-Sup; Yoon, Moon-Young

    2017-01-01

    Overuse of antibiotics has caused serious problems, such as appearance of super bacteria, whose accumulation in the human body through the food chain is a concern. Kanamycin is a common antibiotic used to treat diverse infections; however, residual kanamycin can cause many side effects in humans. Thus, development of an ultra-sensitive, precise, and simple detection system for residual kanamycin in food products is urgently needed for food safety. In this study, we identified kanamycin-binding aptamers via a new screening method, and truncated variants were analyzed for optimization of the minimal sequence required for target binding. We found various aptamers with high binding affinity from 34.7 to 669 nanomolar Kdapp values with good specificity against kanamycin. Furthermore, we developed a reduced graphene oxide (RGO)-based fluorescent aptasensor for kanamycin detection. In this system, kanamycin was detected at a concentration as low as 1 pM (582.6 fg/mL). In addition, this method could detect kanamycin accurately in kanamycin-spiked blood serum and milk samples. Consequently, this simple, rapid, and sensitive kanamycin detection system with newly structural and functional analysis aptamer exhibits outstanding detection compared to previous methods and provides a new possibility for point of care testing and food safety.

  10. Protective role of edaravone against neomycin-induced ototoxicity in zebrafish.

    Science.gov (United States)

    Choi, June; Chang, Jiwon; Jun, Hyung Jin; Im, Gi Jung; Chae, Sung Won; Lee, Seung Hoon; Kwon, Soon-Young; Jung, Hak Hyun; Chung, Ah-Young; Park, Hae-Chul

    2014-05-01

    Aminoglycosides such as neomycin are one of the most commonly prescribed types of antibiotics worldwide. However, these drugs appear to generate free radicals within the inner ear, which can result in permanent hearing loss. We evaluated the effects of edaravone, a neuroprotective agent, on neomycin-induced ototoxicity in transgenic zebrafish. The 5-day post fertilization (dpf) zebrafish larvae were exposed to 125 μM neomycin and various concentrations of edaravone for 1 h. Hair cell survival was calculated as average numbers of the hair cells in the control group, which was not exposed to neomycin. Ultrastructural changes were evaluated using a scanning electron microscope (SEM) and transmission electron microscope (TEM). Edaravone protected against neomycin-induced hair cell loss in the neuromasts (1000 μM: 11.6 ± 1.1 cells, neomycin only: 5.5 ± 0.5 cells; n = 10, Pneomycin and 1000 μM edaravone for 1 h. Edaravone protected against neomycin-induced hair cell loss by preventing apoptosis.

  11. Sequence-specific targeting of RNA with an oligonucleotide-neomycin conjugate.

    Science.gov (United States)

    Charles, Irudayasamy; Xi, Hongjuan; Arya, Dev P

    2007-01-01

    The synthesis of neomycin covalently attached at the C5-position of 2'-deoxyuridine is reported. The synthesis outlined allows for incorporation of an aminoglycoside (neomycin) at any given site in an oligonucleotide (ODN) where a thymidine (or uridine) is present. Incorporation of this modified base into an oligonucleotide, which is complementary to a seven-bases-long alpha-sarcin loop RNA sequence, leads to enhanced duplex hybridization. The increase in Tm for this duplex (DeltaTm = 6 degrees C) suggests a favorable interaction of neomycin within the duplex groove. CD spectroscopy shows that the modified duplex adopts an A-type confirmation. ITC measurements indicate the additive effects of ODN and neomycin binding to the RNA target (Ka = 4.5 x 107 M-1). The enhanced stability of the hybrid duplex from this neomycin-ODN conjugate originates primarily from the enthalpic contribution of neomycin {DeltaDeltaHobs = -7.21 kcal/mol (DeltaHneomycin conjugated - DeltaH nonconjugated)} binding to the hybrid duplex. The short linker length allows for selective stabilization of the hybrid duplex over the hybrid triplex. The results described here open up new avenues in the design and synthesis of nucleo-aminoglycoside-conjugates (N-Ag-C) where the inclusion of any number of aminoglycoside (neomycin) molecules per oligonucleotide can be accomplished.

  12. Effect of neomycin and protein S1 on the binding of streptomycin to the ribosome.

    Science.gov (United States)

    Grisé-Miron, L; Brakier-Gingras, L

    1982-04-01

    The binding of [3H]dihydrostreptomycin to the 70-S ribosome or to the 30-S subunit has been investigated in the presence of neomycin by the Millipore filtration or the equilibrium dialysis procedure. It was observed that dihydrostreptomycin binds equally well to the 30-S subunit and the 70-S ribosome, and that neomycin stimulates the binding of dihydrostreptomycin to the ribosome by increasing the association constant and not by creating new binding sites. Specific removal of protein S1 from the 30-S subunit neither affected the binding of dihydrostreptomycin to the ribosome nor the stimulation of dihydrostreptomycin binding by neomycin.

  13. 2,3-Dihydroxybenzoic acid attenuates kanamycin-induced volume reduction in mouse utricular type I hair cells

    DEFF Research Database (Denmark)

    Severinsen, Stig Åvall; Kirkegaard, Mette; Nyengaard, Jens Randel

    2006-01-01

    The aminoglycoside kanamycin is a commonly used antibiotic, but unfortunately it is oto- and nephrotoxic in large doses. The negative effects are thought to be due to the formation of free radicals which is why strong antioxidants and iron chelators like 2,3-dihydroxybenzoic acid (DHB) are of great...... interest. This study estimates cellular quantitative changes in the utricular macula of mice following systemic treatment with kanamycin alone or in combination with DHB. The animals were injected with either saline, kanamycin or kanamycin+DHB for 15 days and perfusion fixed three weeks after last...... macula, hair cell type I and supporting cells decreased significantly in animals injected with kanamycin but not in animals co-treated with DHB. Hair and supporting cell numbers remained unchanged in all three groups. In conclusion, the kanamycin-induced volume reduction of type I hair cells...

  14. Label-free detection of kanamycin based on a G-quadruplex DNA aptamer-based fluorescent intercalator displacement assay

    Science.gov (United States)

    Xing, Yun-Peng; Liu, Chun; Zhou, Xiao-Hong; Shi, Han-Chang

    2015-01-01

    This work was the first to report that the kanamycin-binding DNA aptamer (5'-TGG GGG TTG AGG CTA AGC CGA-3') can form stable parallel G-quadruplex DNA (G4-DNA) structures by themselves and that this phenomenon can be verified by nondenaturing polyacrylamide gel electrophoresis and circular dichroism spectroscopy. Based on these findings, we developed a novel label-free strategy for kanamycin detection based on the G4-DNA aptamer-based fluorescent intercalator displacement assay with thiazole orange (TO) as the fluorescence probe. In the proposed strategy, TO became strongly fluorescent upon binding to kanamycin-binding G4-DNA. However, the addition of kanamycin caused the displacement of TO from the G4-DNA-TO conjugate, thereby resulting in decreased fluorescent signal, which was inversely related to the kanamycin concentration. The detection limit of the proposed assay decreased to 59 nM with a linear working range of 0.1 μM to 20 μM for kanamycin. The cross-reactivity against six other antibiotics was negligible compared with the response to kanamycin. A satisfactory recovery of kanamycin in milk samples ranged from 80.1% to 98.0%, confirming the potential of this bioassay in the measurement of kanamycin in various applications. Our results also served as a good reference for developing similar fluorescent G4-DNA-based bioassays in the future.

  15. ESTABLISHMENT OF PHELIPANCHE RAMOSA TISSUE CULTURE AND EFFECT OF KANAMYCIN ON CULTURE GROWTH

    Directory of Open Access Journals (Sweden)

    Dagmara Kullačová

    2015-02-01

    Full Text Available Orobanchaceae family includes parasitic plants that attack many important food crops. Genus Phelipanche, belonging to this family is considered to cause high negative impact on food production. Developing Phelipanche plant must establish connection with the root of host plant, from which it receives all resources needed for further development. Nowadays big effort is directed to finding a reliable strategy to control parasitic plants. In vitro cultures of P. ramosa can be genetically manipulated and used for study of genes involved in host-parasite interactions. We established in vitro cultures of parasitic species Phelipanche ramosa on solid and liquid media in parallel. The obtained results point out that development of P. ramosa calli was origin specific. We tested the effect of antibiotic kanamycin on in vitro cultures of Phelipanche ramosa with aim to develop system for its genetic manipulation and selection of transgenic tissue using kanamycin- resistance approach. The selection pressure of kanamycin was stronger in liquid grown cultures. However, concentrations of kanamycin tested (up to 250 mg.l-1 did not ensured elimination of kanamycine non-resistant tissue. Tests of other candidate selection markers are currently in progress.

  16. Colorimetric and fluorometric detection of neomycin based on conjugated polydiacetylene supramolecules.

    Science.gov (United States)

    Zhou, Guodong; Wang, Fang; Wang, Huilin; Kambam, Srinivasulu; Chen, Xiaoqiang

    2013-06-13

    Utilizing the colorimetric and fluorogenic changes, a system based on polydiacetylenes (PDAs) is developed for the detection of neomycin. The PDA supramolecules polymerized from the mixed liposome composed of N-(3-hydroxyphenyl)pentacosa-10,12-diynamide (PCDA-AP) and pentacosa-10,12-diynoic acid (PCDA) at an optimized ratio of 1:9 display a unique colorimetric change (blue to red) and fluorescent enhancement in the presence of neomycin. The detection limit for neomycin is estimated to be 2.55 × 10(-7) M by the fluorogenic method. The optical changes induced by neomycin can be attributed to the disruption of the hydrogen bonding between phenol and carboxylic acid from PCDA-AP and PCDA.

  17. Molecular recognition of single-stranded RNA: neomycin binding to poly(A).

    Science.gov (United States)

    Xi, Hongjuan; Gray, David; Kumar, Sunil; Arya, Dev P

    2009-07-01

    Poly(A) is a relevant sequence in cell biology due to its importance in mRNA stability and translation initiation. Neomycin is an aminoglycoside antibiotic that is well known for its ability to target various nucleic acid structures. Here it is reported that neomycin is capable of binding tightly to a single-stranded oligonucleotide (A(30)) with a K(d) in the micromolar range. CD melting experiments support complex formation and indicate a melting temperature of 47 degrees C. The poly(A) duplex, which melts at 44 degrees C (pH 5.5), was observed to melt at 61 degrees C in the presence of neomycin, suggesting a strong stabilization of the duplex by the neomycin.

  18. Semi-solid-state fermentation: a promising alternative for neomycin production by the actinomycete Streptomyces fradiae.

    Science.gov (United States)

    Machado, Isabel; Teixeira, José A; Rodríguez-Couto, Susana

    2013-06-10

    The production of neomycin by the actinomycete Streptomyces fradiae, under semi-solid-state fermentation conditions was the main subject of this study. Two supports (nylon sponge and orange peelings) were tested in order to determine the most suitable one for the production of neomycin by the above-mentioned microorganism. Nylon sponge led to the highest neomycin production, reaching a maximum value of 13,903 μg/mL on the 10th day of cultivation. As a control, the same experiment was performed under submerged fermentation (SmF) conditions, without solid support. Here the production of neomycin by S. fradiae was about 55-fold lower (i.e. 250 μg/mL) than that obtained for SSF.

  19. Collaborative study for the establishment of the 3rd international standard for neomycin.

    Science.gov (United States)

    Rautmann, G; Daas, A; Buchheit, K-H

    2013-01-01

    An international collaborative study was organised to establish the World Health Organization (WHO) 3(rd) International Standard (IS) for neomycin. Ten laboratories from different countries participated in the collaborative study. The potency of the candidate material, a freeze-dried preparation, was estimated by microbiological assays with sensitive micro-organisms. To ensure continuity between consecutive batches, the 2(nd) IS for neomycin was used as a standard. Based on the results of the study, the 3(rd) IS for neomycin was adopted at the meeting of the WHO Expert Committee on Biological Standardization (ECBS) in 2012 with an assigned potency of 19,050 IU per vial. The 3(rd) IS for neomycin is available from the European Directorate for the Quality of Medicines & HealthCare (EDQM).

  20. Oral administration of neomycin to chickens experimentally infected with Salmonella typhimurium.

    Science.gov (United States)

    Smith, H W; Tucker, J F

    1978-04-22

    Groups of healthy chickens with a light experimental Salmonella typhimurium infection were fed on a diet containing 225 g per ton (1016 kg) of neomycin for two days. This brought about only a slight reduction in the incidence of chickens that were excreting S typhimurium in their faeces. Examination of caecal contents two days after the cessation of treatment revealed the neomycin had not had any effect in eliminating infection. In one experiment, the neomycin administration resulted in the emergence of enormous populations of Escherichia coli in the alimentary tract of treated chickens that possessed multiple antibiotic resistance of the transmissible type. For these reasons the practice of feeding broiler chickens on diets containing neomycin immediately before slaughter should be actively discouraged.

  1. Modulation of irinotecan-induced diarrhea by cotreatment with neomycin in cancer patients

    NARCIS (Netherlands)

    D.F.S. Kehrer (Diederik); A. Sparreboom (Alex); J. Verweij (Jaap); P. de Bruijn (Peter); C.A. Nierop; J. van de Schraaf; E.J. Ruijgrok (Elisabeth); M.J.A. de Jonge (Maja)

    2001-01-01

    textabstractThis study was designed to evaluate irinotecan (CPT-11) disposition and pharmacodynamics in the presence and absence of the broad-spectrum antibiotic neomycin. Seven evaluable cancer patients experiencing diarrhea graded > or =2 after receiving CPT-11 alone

  2. Comparison of the action of streptomycin and neomycin on the structure of the bacterial ribosome.

    Science.gov (United States)

    Noreau, J; Grisé-Miron, L; Brakier-Gingras, L

    1980-06-27

    The influence of streptomycin and neomycin upon the conformation of the ribosome has been investigated using spin-labeled and fluorescent analogs of the sulfhydryl reagent, N-ethylmaleimide. Changes in the electron paramagnetic resonance spectra or in the polarization of fluorescence of labeled ribosomes reveal that streptomycin alters the mobility of labels bound to the sulfhydryl group of protein S18 while neomycin affects the mobility of labels bound to the sulfhydryl groups of proteins S1, S21 and/or L10. It is also observed that both streptomycin and neomycin interfere with changes in the mobility of labels induced by storage under inactivating conditions. From these results, it is concluded that: 1. streptomycin and neomycin distort the conformation of the ribosome at different sites, streptomycin disturbing preferentially the area around the sulfhydryl group of protein S18 while neomycin affects the environment of the sulfhydryl groups of proteins S1, S21 and/or L10; 2. streptomycin and neomycin interefere with the ability of the ribosome to undergo conformational changes.

  3. Effects of the aminoglycoside antibiotics, streptomycin and neomycin, on neuromuscular transmission. II. Postsynaptic considerations.

    Science.gov (United States)

    Fiekers, J F

    1983-06-01

    The postsynaptic effects of two aminoglycoside antibiotics, streptomycin and neomycin, were studied on miniature end-plate currents (mepcs) and acetylcholine-induced end-plate current fluctuations in voltage-clamped costocutaneous muscles of the garter snake (species Thamnophis). Neomycin decreased the amplitude of mepcs and accelerated the time constants of mepc decay in a concentration-dependent manner without altering the single exponential nature of mepc decay. Neomycin also produced a voltage- and concentration-dependent nonlinearity in the current/voltage relationship. The relationship between the time constants of mepc decay and membrane potential was progressively reduced with increasing concentrations of neomycin. A concentration-dependent reduction in single channel conductance and channel lifetime was also obtained with neomycin. In contrast, streptomycin, in concentrations up to 5 X 10(-5) M, did not significantly alter either mepc amplitude, the time constant of mepc decay, the relationship between the mepc decay time constant and membrane potential or the lifetime and conductance of single end-plate channels. In very high concentrations (greater than 1 mM) streptomycin decreased mepc amplitude and prolonged mepc decay at hyperpolarized membrane potentials. The results suggest that neomycin interacts with the ionic channels of the acetylcholine receptor in their open configuration, whereas streptomycin acts primarily by blocking the receptor. The significant differences in the molecular actions of these two antibiotics may provide an explanation for the observed differences in the character and reversal of the neuromuscular block produced by these antibiotics.

  4. Protective role of NecroX-5 against neomycin-induced hair cell damage in zebrafish.

    Science.gov (United States)

    Song, Jae-Jun; Chang, Jiwon; Choi, Jungim; Im, Gi Jung; Chae, Sung Won; Lee, Seung Hoon; Kwon, Soon-Young; Jung, Hak Hyun; Chung, Ah-Young; Park, Hae-Chul; Choi, June

    2014-02-01

    NecroX-5, one of the derivatives of NecroX series compounds, is a mitochondrial reactive oxygen species and reactive nitrogen species scavenger that inhibits cell death against various kinds of oxidative stresses. The objective of the present study was to evaluate the effects of NecroX-5 on neomycin-induced ototoxicity in transgenic zebrafish (Brn3C: EGFP). Five days post-fertilization, zebrafish larvae were exposed to 125 μM neomycin and one of the following NecroX-5 concentrations for 1 h: 10, 25, 50, and 75 μM. Hair cells within the neuromasts of the supraorbital (SO1 and SO2), otic (O1), and occipital (OC1) lateral lines were analyzed using fluorescence microscopy (n = 10). The terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay and 2-[4-(dimethylamino) styryl]-N-ethylpyridiniumiodide (DASPEI) assay were performed for evaluation of apoptosis and mitochondrial damage. Ultrastructural changes were evaluated using scanning electron microscopy. NecroX-5 decreased neomycin-induced hair cell loss in the neuromasts (NecroX-5 50 μM: 13.4 ± 2.0 cells, 125 μM neomycin only: 8.1 ± 1.2 cells; n = 10, P neomycin and 50 μM NecroX-5. NecroX-5 decreased apoptosis and mitochondrial damage. In conclusion, NecroX-5 attenuated neomycin-induced hair cell loss in zebrafish.

  5. Neomycin-neomycin dimer: an all-carbohydrate scaffold with high affinity for AT-rich DNA duplexes.

    Science.gov (United States)

    Kumar, Sunil; Xue, Liang; Arya, Dev P

    2011-05-18

    A dimeric neomycin-neomycin conjugate 3 with a flexible linker, 2,2'-(ethylenedioxy)bis(ethylamine), has been synthesized and characterized. Dimer 3 can selectively bind to AT-rich DNA duplexes with high affinity. Biophysical studies have been performed between 3 and different nucleic acids with varying base composition and conformation by using ITC (isothermal calorimetry), CD (circular dichroism), FID (fluorescent intercalator displacement), and UV (ultraviolet) thermal denaturation experiments. A few conclusions can be drawn from this study: (1) FID assay with 3 and polynucleotides demonstrates the preference of 3 toward AT-rich sequences over GC-rich sequences. (2) FID assay and UV thermal denaturation experiments show that 3 has a higher affinity for the poly(dA)·poly(dT) DNA duplex than for the poly(dA)·2poly(dT) DNA triplex. Contrary to neomycin, 3 destabilizes poly(dA)·2poly(dT) triplex but stabilizes poly(dA)·poly(dT) duplex, suggesting the major groove as the binding site. (3) UV thermal denaturation studies and ITC experiments show that 3 stabilizes continuous AT-tract DNA better than DNA duplexes with alternating AT bases. (4) CD and FID titration studies show a DNA binding site size of 10-12 base pairs/drug, depending upon the structure/sequence of the duplex for AT-rich DNA duplexes. (5) FID and ITC titration between 3 and an intramolecular DNA duplex [d(5'-A(12)-x-T(12)-3'), x = hexaethylene glycol linker] results in a binding stoichiometry of 1:1 with a binding constant ∼10(8) M(-1) at 100 mM KCl. (6) FID assay using 3 and 512 hairpin DNA sequences that vary in their AT base content and placement also show a higher binding selectivity of 3 toward continuous AT-rich than toward DNA duplexes with alternate AT base pairs. (7) Salt-dependent studies indicate the formation of three ion pairs during binding of the DNA duplex d[5'-A(12)-x-T(12)-3'] and 3. (8) ITC-derived binding constants between 3 and DNA duplexes have the following order: AT

  6. INFLUENCE OF NEOMYCIN AND INGESTED ENDOTOXIN IN THE PATHOGENESIS OF CHOLINE DEFICIENCY CIRRHOSIS IN THE ADULT RAT

    Science.gov (United States)

    Broitman, Selwyn A.; Gottlieb, Leonard S.; Zamcheck, Norman

    1964-01-01

    Two groups of adult rats fed a choline-deficient diet supplemented with neomycin in their drinking water for 250 or 350 days were protected against the development of liver fibrosis and cirrhosis. At the termination of the study these animals weighed more than others not receiving neomycin. This difference in weight did not appear to be caused by a growth-promoting effect of neomycin but rather reflected the increased severity of liver disease and a resultant weight loss in animals not receiving neomycin. Protection by neomycin was cancelled when Salmonella typhosa endotoxin was added to the drinking water. It was concluded that the protective effect of neomycin was mediated by an alteration in the intestinal microflora resulting in a reduction in the numbers of organisms contributing to intraluminal endotoxin. In the presence of choline deficiency, absorption of intraluminal endotoxin may contribute to the development of fibrosis and cirrhosis. PMID:14151103

  7. Quantification of amikacin and kanamycin in serum using a simple and validated LC-MS/MS method

    NARCIS (Netherlands)

    Dijkstra, Jacob A.; Sturkenboom, Marieke G. G.; van Hateren, Kai; Koster, Remco A.; Greijdanus, Ben; Alffenaar, Jan-Willem C.

    2014-01-01

    Background: Amikacin and kanamycin are frequently used in the treatment of multidrug-resistant TB. The current commercially available immunoassay is unable to analyze kanamycin and trough levels of amikacin. The objective was therefore to develop a LC-MS/MS method for the quantification of amikacin

  8. Design, synthesis, and antibacterial activities of neomycin-lipid conjugates: polycationic lipids with potent gram-positive activity.

    Science.gov (United States)

    Bera, Smritilekha; Zhanel, George G; Schweizer, Frank

    2008-10-01

    Aminoglycoside antibiotics and cationic detergents constitute two classes of clinically important drugs and antiseptics. Their bacteriological and clinical efficacy, however, has decreased recently due to antibiotic resistance. We have synthesized aminoglycoside-lipid conjugates in which the aminoglycoside neomycin forms the cationic headgroup of a polycationic detergent. Our results show that neomycin-C16 and neomycin-C20 conjugates exhibit strong Gram-positive activity but reduced Gram-negative activity. The MIC of neomycin-C16 (C20) conjugates against methicillin-resistant Staphylococcus aureus (MRSA) is comparable to clinically used antiseptics.

  9. Novel plasmid conferring kanamycin and tetracycline resistance in turkey-derived Campylobacter jejuni strain 11601MD

    Science.gov (United States)

    In Campylobacter spp., resistance to the antibiotics kanamycin and tetracycline is frequently associated with plasmid-borne genes. However, relatively few plasmids of Campylobacter jejuni have been fully characterized to date. A novel plasmid (p11601MD; 44,095 bp.) harboring tet(O) was identified in...

  10. Screening of Transgenic Soybean Transformed by Means of Pollen-tube Using Kanamycin

    Institute of Scientific and Technical Information of China (English)

    ZHANG Shuzhen; XU Pengfei; ZHANG Dayong; LIN Shifeng; LI Wenbin; HAN Yingpeng; YANG Chuanping

    2006-01-01

    Kanamycin was used to screen T0 seeds of the variety Dongnong 46 transformed by means of and 2 positive plants were gotten combined with Gus dyeing and PCR detection. It is proved that this method is economic and effective in preliminary screening the transgenic plants.

  11. Interaction of weakly bound antibiotics neomycin and lincomycin with bovine and human serum albumin: biophysical approach.

    Science.gov (United States)

    Keswani, Neelam; Choudhary, Sinjan; Kishore, Nand

    2010-07-01

    The thermodynamics of interaction of neomycin and lincomycin with bovine serum albumin (BSA) and human serum albumin (HSA) has been studied using isothermal titration calorimetry (ITC), in combination with UV-visible, steady state and time resolved fluorescence spectroscopic measurements. Neomycin is observed to bind weakly to BSA and HSA whereas lincomycin did not show any evidence for binding with the native state of these proteins, rather it interacts in the presence of surfactants. The ITC results suggest 1 : 1 binding stoichiometry for neomycin in the studied temperature range. The values of the van't Hoff enthalpy do not agree with the calorimetric enthalpy in the case of neomycin, suggesting conformational changes in the protein upon ligand binding, as well as with the rise in the temperature. Experiments at different ionic strengths, and in the presence of tetrabutyl ammonium bromide and surfactants suggest the predominant involvement of electrostatic interactions in the complexation process of neomycin with BSA and HSA, and non-specific interaction behaviour of lincomycin with these proteins.

  12. Effect of covalent attachment of neomycin on conformational and aggregation properties of catalase.

    Science.gov (United States)

    Hashemnia, S; Mokhtari, Z; Tashkhourian, J; Moosavi-Movahedi, A A

    2015-04-01

    The carboxylic groups of glutamic acid and aspartic acid residues of catalase (CAT) were chemically modified using the treatment of the enzyme with 1-ethyl-3-(3'-dimethylamino) carbodiimide hydrochloride (EDC) and neomycin. The effect of covalent attachment of neomycin on the enzymatic activity, conformational and aggregation properties of CAT was investigated. The modification of CAT with different concentrations of neomycin showed two different types of behavior, depending up on the concentration range of neomycin. In the concentration range from 0.0 to 5.2 mM, neomycin-modified CAT, compared to the native enzyme exhibited higher a-helix content, reduced surface hydrophobicity, little enhancement in CAT activity and a better protection against thermal aggregation, whereas at concentrations greater than 5.2 mM, the modified enzyme exhibited a significant decrease in CAT activity and an increase in random coil content which may result in disorder in the protein structure and increase in thermal aggregation. This modification is a rapid and simple approach to investigate the role of aspartate and glutamate residues in the structure, function and folding of CAT.

  13. Correlation of neomycin, faecal neutral and acid sterols with colon carcinogenesis in rats.

    Science.gov (United States)

    Panda, S K; Chattoraj, S C; Broitman, S A

    1999-06-01

    High fat diets have been implicated in incidence of colon cancer both in epidemiological and animal studies. Present investigation deals with the incidence, location and numbers of large and small bowel tumours induced by 1,2-dimethyl hydrazine (DMH) in rats fed high fat diets and neomycin. Neomycin was used to modify the faecal sterol metabolism and the relationship of the high fat diet and faecal neutral and acid sterols to the large bowel tumorigenesis was evaluated. DMH administered rats were fed with (a) 20% safflower oil; (b) 20% safflower oil and neomycin; (c) 20% safflower oil, cholesterol and cholic acid; and (d) 20% safflower oil, cholesterol, cholic acid and neomycin. Neomycin was found to be associated with both increase and decrease of tumour numbers. The faecal sterols lithocholic and deoxycholic acids were found to have no participation, while cholesterol and cholic acid were found to decrease with increase in tumour numbers. However, faecal coprostanol has been found to have a significant positive correlation with tumorigenesis in all dietary groups. Therefore coprostanol might possibly be associated with colon carcinogenesis in DMH-fed rats and cholesterol metabolism in gut appears to be related to the development of tumours.

  14. The antibiotic neomycin abolishes directional selectivity in rabbit retinal ganglion cells.

    Science.gov (United States)

    Jensen, R J

    1996-12-01

    1. Extracellular recordings from ON/OFF directionally selective ganglion cells in superfused rabbit retinas were made to study the effect of the aminoglycoside antibiotic, neomycin, on the responses of these cells to a moving light stimulus. 2. Neomycin, at 480-800 microM, reversibly abolished the directional selectivity in these ganglion cells by bringing out a response to movement in one ("null") direction that was similar in magnitude to the response to movement in the reverse ("preferred") direction. 3. Gentamicin, streptomycin, and tobramycin were also able to abolish directional selectivity in these ganglion cells but only at concentrations greater than 1000 microM. 4. It is proposed that neomycin abolishes directional selectivity in rabbit retinal ganglion cells by blocking omega-conotoxin MVIIC-sensitive Ca2+ channels in the retina.

  15. Determination of neomycin in water samples by high performance anion chromatography with pulsed amperometric detection

    Institute of Scientific and Technical Information of China (English)

    Bin Guan; Dong Xing Yuan

    2007-01-01

    A simple, fast and reliable method, using high performance anion chromatography with pulsed amperometric detection, had been developed for the analysis of neomycin in water samples. The elution and separation were carried out with an isocratic mobile phase, containing 10 mmol/L NaOH. The influence of the concentration and pH of the mobile phase on the separation and detection was investigated. A quadruple-potential waveform used for the detection was optimized. The detection limit of neomycin was down to 0.027 μg/mL. The linearity of neomycin calibration curve ranged from 0.050 to 0.505 μg/mL with correlation coefficient of0.9997. R.S.D. (n= 11) was 4.0%.

  16. Inhibition of H3K4me2 Demethylation Protects Auditory Hair Cells from Neomycin-Induced Apoptosis.

    Science.gov (United States)

    He, Yingzi; Yu, Huiqian; Cai, Chengfu; Sun, Shan; Chai, Renjie; Li, Huawei

    2015-08-01

    Aminoglycoside-induced hair cell loss is a major cause of hearing impairment in children and deserves more attention in medical research. Epigenetic mechanisms have been shown to protect hair cells from ototoxic drugs. In this study, we focused on the role of dimethylated histone H3K4 (H3K4me2) in hair cell survival. To investigate the effects of lysine-specific demethylase 1 (LSD1)--the histone demethylase primarily responsible for demethylating H3K4me2--on neomycin-induced hair cell loss, isolated cochleae were pretreated with LSD1 inhibitors followed by neomycin exposure. There was a severe loss of hair cells in the organ of Corti after neomycin exposure, and inhibition of LSD1 significantly protected against neomycin-induced hair cell loss. H3K4me2 expression in the nuclei of hair cells decreased after exposure to neomycin, and blocking the decreased expression of H3K4me2 with LSD1 inhibitors prevented hair cell loss. Local delivery of these inhibitors in vivo also protected hair cells from neomycin-induced ototoxicity and maintained the hearing threshold in mice as determined by auditory brain stem response. This inhibition of neomycin-induced apoptosis occurs via reduced caspase-3 activation. Together, our findings demonstrate the protective role for H3K4me2 against neomycin-induced hair cell loss and hearing loss.

  17. Neomycin inhibits histamine and thapsigargin mediated Ca2+ DDT1 MF-2 cells independent of phospholipase C activation

    NARCIS (Netherlands)

    Sipma, H; VanderZee, L; DenHertog, A; Nelemans, A

    1996-01-01

    The histamine H-1 receptor mediated increase in cytoplasmic Ca2+ ([Ca2+](i)) was measured in the presence of the known phospholipase C (PLC) inhibitor, neomycin. Neomycin (1 mM) inhibited the histamine (100 mu M) induced rise in [Ca2+](i) to the same extent as observed after blocking Ca2+ entry with

  18. Global epigenetic changes induced by SWI2/SNF2 inhibitors characterize neomycin-resistant mammalian cells.

    Directory of Open Access Journals (Sweden)

    Popy Dutta

    Full Text Available BACKGROUND: Previously, we showed that aminoglycoside phosphotransferases catalyze the formation of a specific inhibitor of the SWI2/SNF2 proteins. Aminoglycoside phosphotransferases, for example neomycin-resistant genes, are used extensively as selection markers in mammalian transfections as well as in transgenic studies. However, introduction of the neomycin-resistant gene is fraught with variability in gene expression. We hypothesized that the introduction of neomycin-resistant genes into mammalian cells results in inactivation of SWI2/SNF2 proteins thereby leading to global epigenetic changes. METHODOLOGY: Using fluorescence spectroscopy we have shown that the inhibitor, known as Active DNA-dependent ATPase ADomain inhibitor (ADAADi, binds to the SWI2/SNF2 proteins in the absence as well as presence of ATP and DNA. This binding occurs via a specific region known as Motif Ia leading to a conformational change in the SWI2/SNF2 proteins that precludes ATP hydrolysis. ADAADi is produced from a plethora of aminoglycosides including G418 and Streptomycin, two commonly used antibiotics in mammalian cell cultures. Mammalian cells are sensitive to ADAADi; however, cells stably transfected with neomycin-resistant genes are refractory to ADAADi. In resistant cells, endogenous SWI2/SNF2 proteins are inactivated which results in altered histone modifications. Microarray data shows that the changes in the epigenome are reflected in altered gene expression. The microarray data was validated using real-time PCR. Finally, we show that the epigenetic changes are quantized. SIGNIFICANCE: The use of neomycin-resistant genes revolutionized mammalian transfections even though questions linger about efficacy. In this study, we have demonstrated that selection of neomycin-resistant cells results in survival of only those cells that have undergone epigenetic changes, and therefore, data obtained using these resistant genes as selection markers need to be cautiously

  19. Neomycin and carbodiimide crosslinking as an alternative to glutaraldehyde for enhanced durability of bioprosthetic heart valves.

    Science.gov (United States)

    Leong, Joshua; Munnelly, Amy; Liberio, Brianna; Cochrane, Leonard; Vyavahare, Naren

    2013-05-01

    Glutaraldehyde cross-linked porcine aortic valves, referred to as bioprosthetic heart valves (BHVs), are often used in heart valve replacements. Glutaraldehyde does not stabilize glycosaminoglycans (GAGs) and they are lost during preparation, in vivo implantation, cyclic fatigue, and storage. We report that binding of neomycin, a hyaluronidase inhibitor, to the tissues with carbodiimide cross-linking improves GAG retention without reducing collagen and elastin stability. It also led to improved biomechanical properties. Neomycin carbodiimide cross-linking did not significantly reduce calcification in a rat subdermal implantation model when they were stored in formaldehyde after cross-linking. Removal of formaldehyde storage significantly reduced calcification.

  20. Dual recognition of the human telomeric G-quadruplex by a neomycin-anthraquinone conjugate.

    Science.gov (United States)

    Ranjan, Nihar; Davis, Erik; Xue, Liang; Arya, Dev P

    2013-06-28

    The authors report the recognition of a G-quadruplex formed by four repeat human telomeric DNA with aminosugar intercalator conjugates. The recognition of the G-quadruplex through dual binding mode ligands significantly increased the affinity of ligands for the G-quadruplex. One such example is a neomycin-anthraquinone conjugate (2) which exhibited nanomolar affinity for the quadruplex, and the affinity of (2) is nearly 1000 fold higher for the human telomeric G-quadruplex DNA than its constituent units, neomycin and anthraquinone.

  1. Characterization of a radical S-adenosyl-L-methionine epimerase, NeoN, in the last step of neomycin B biosynthesis.

    Science.gov (United States)

    Kudo, Fumitaka; Hoshi, Shota; Kawashima, Taiki; Kamachi, Toshiaki; Eguchi, Tadashi

    2014-10-01

    The last step of neomycin biosynthesis is the epimerization at C-5‴ of neomycin C to give neomycin B. A candidate enzyme responsible for the epimerization was a putative radical S-adenosyl-L-methionine (SAM) enzyme, NeoN, which is uniquely encoded in the neomycin biosynthetic gene cluster and remained an unassigned protein in the neomycin biosynthesis. The reconstituted and reduced NeoN showed the expected epimerization activity in the presence of SAM. In the epimerization, 1 equiv of SAM was consumed to convert neomycin C into neomycin B. The site of neomycin C reactive toward epimerization was clearly confirmed to be C-5‴ by detecting the incorporation of a deuterium atom from the deuterium oxide-based buffer solution. Further, alanine scanning of the NeoN cysteine residues revealed that C249 is a critical amino acid residue that provides a hydrogen atom to complete the epimerization. Furthermore, electron paramagnetic resonance analysis of the C249A variant in the presence of SAM and neomycin C revealed that a radical intermediate is generated at the C-5‴ of neomycin C. Therefore, the present study clearly illustrates that the epimerization of neomycin C to neomycin B is catalyzed by a unique radical SAM epimerase NeoN with a radical reaction mechanism.

  2. Controversy Associated With the Common Component of Most Transgenic Plants – Kanamycin Resistance Marker Gene

    OpenAIRE

    Jelenić, Srećko

    2003-01-01

    Plant genetic engineering is a powerful tool for producing crops resistant to pests, diseases and abiotic stress or crops with improved nutritional value or better quality products. Currently over 70 genetically modified (GM) crops have been approved for use in different countries. These cover a wide range of plant species with significant number of different modified traits. However, beside the technology used for their improvement, the common component of most GM crops is the neomycin phosp...

  3. Report concerning the colaborative study for establish of reference standard eficacity of Kanamycin

    Directory of Open Access Journals (Sweden)

    Simona Sturzu

    2015-12-01

    Full Text Available The Microbiology Laboratory from the Institute for the Control of Veterinary Biological Products and Medicines participated to a collaborative study in order to determine the potency of kanamycin CRS, batch 3, according to the protocol sent by the EDQM (European Directorate for the Quality of Medicines - coordinating of the study. The purpose of the study was to establish the potency of the batch 3 for in the characterization of the substance as reference standard. Potency was determined by microbiological method, based on comparison of the inhibition zones of growth of micro-organisms sensitive to those of a reference standard, as specified in European Pharmacopoeia, Chapter 2.7.2. - Microbiological testing of antibiotics. After analysis of received data from participants and their statistical processing by the EDQM, the potency of the batch 3, kanamycin - reference materials, was established in 790 UI/mg.

  4. MAPLE fabrication of thin films based on kanamycin functionalized magnetite nanoparticles with anti-pathogenic properties

    Science.gov (United States)

    Grumezescu, Valentina; Andronescu, Ecaterina; Holban, Alina Maria; Mogoantă, Laurenţiu; Mogoşanu, George Dan; Grumezescu, Alexandru Mihai; Stănculescu, Anca; Socol, Gabriel; Iordache, Florin; Maniu, Horia; Chifiriuc, Mariana Carmen

    2015-05-01

    In this study we aimed to evaluate the biocompatibility and antimicrobial activity of kanamycin functionalized 5 nm-magnetite (Fe3O4@KAN) nanoparticles thin films deposited by Matrix Assisted Pulsed Laser Evaporation (MAPLE) technique. A laser deposition regime was established in order to stoichiometrically transfer Fe3O4@KAN thin films on silicone and glass substrates. Morphological and physico-chemical properties of powders and coatings were characterized by XRD, TEM, SEM, AFM and IR microscopy (IRM). Our nanostructured thin films have proved efficiency in the prevention of microbial adhesion and mature biofilms development as a result of antibiotic release in its active form. Furthermore, kanamycin functionalized nanostructures exhibit a good biocompatibility, both in vivo and in vitro, demonstrating their potential for implants application. This is the first study reporting the assessment of the in vivo biocompatibility of a magnetite-antimicrobial thin films produced by MAPLE technique.

  5. EDTA-dependent pseudothrombocytopenia confirmed by supplementation of kanamycin; a case report.

    Science.gov (United States)

    Ahn, Hae Lyun; Jo, Young Il; Choi, Young Suk; Lee, Jung Yeon; Lee, Hae Woon; Kim, Seong Ryul; Sim, Joon; Lee, Weon; Jin, Chun Jo

    2002-03-01

    EDTA-dependent pseudothrombocytopenia (PTCP) is the phenomenon of a spurious low platelet count due to EDTA-induced aggregation of platelets. Since the failure to recognize EDTA-dependent PTCP may result in incorrect diagnosis and inappropriate treatment, the recognition of this phenomenon is very important. We report an insidious case of EDTA-dependent PTCP confirmed by supplementation of kanamycin to anticoagulant in a 53-year-old women. Although sodium citrate and heparin usually prevented the aggregation of platelets in EDTA-dependent PTCP patients, these anticoagulants failed in preventing PTCP in our case. EDTA-dependent PTCP was confirmed by the findings that the clumping of platelets on microscopic evaluation was found in EDTA-anticoagulated blood samples, whereas thrombocytopenia and platelet aggregation were not revealed in the sample supplemented with kanamycin.

  6. Evaluation of genetic mutations associated with Mycobacterium tuberculosis resistance to amikacin, kanamycin and capreomycin: a systematic review

    National Research Council Canada - National Science Library

    Georghiou, Sophia B; Magana, Marisa; Garfein, Richard S; Catanzaro, Donald G; Catanzaro, Antonino; Rodwell, Timothy C

    2012-01-01

    ...%. While this relationship is well established and reliable for first-line anti-TB drugs, rifampin and isoniazid, it is less well-studied and understood for second-line, injectable drugs, amikacin (AMK), kanamycin (KAN) and capreomycin (CAP...

  7. Management of gonococcal ophthalmia neonatorum with single-dose kanamycin and ocular irrigation with saline.

    Science.gov (United States)

    Latif, A; Mason, P; Marowa, E; Paraiwa, E; Dhamu, F; Tambo, J; Gwanzura, L; Mapeta, D; Jongeling, G

    1988-01-01

    Two hundred nineteen neonates with gonococcal ophthalmia neonatorum, including 40 infected with penicillinase-producing strains, were treated as outpatients with a single intramuscular injection of 100 mg of kanamycin and hourly ocular irrigation with saline. Neisseria gonorrhoeae was isolated from three (1.4%) of the 212 babies attending for follow-up, and post-gonococcal conjunctivitis developed in 22 (10.4%) of those who returned for follow-up.

  8. RENAL CLEARANCE AND URINARY EXCRETION OF KANAMYCIN IN DOMESTIC RUMINANT SPECIES

    Directory of Open Access Journals (Sweden)

    I. JAVED, Z. U. RAHMAN, F. H. KHAN, F. MUHAMMAD, Z. IQBAL AND B. ASLAM

    2006-01-01

    Full Text Available Species dependent geonetical differences in renal clearance and urinary excretion of kanamycin were investigated in adult female buffaloes, cows, sheep and goats. The drug was administered as a single intravenous dose (5 mg/kg b.wt. Blood and urine samples were collected at various time intervals after drug administration. The plasma and urine concentrations of the drug were determined using the microbiological assay. The mean (± SE values for endogenous creatinine clearance (an index of glomerular filtration rate were 0.77 ± 0.05, 0.49 ± 0.07, 0.81 ± 0.07 and 0.98 ± 0.13 ml/min.kg in buffaloes, cows, sheep and goats, respectively. Experiments regarding kidney handling of kanamycin in these ruminant species revealed respective values of renal clearance as 0.08 ± 0.01, 0.07 ± 0.01, 0.19 ± 0.02 and 0.23 ± 0.04 ml/min.kg. Besides glomerular filtration, kanamycin was reabsorbed from the renal tubules of all ruminant species and actively secreted into the renal tubules of buffaloes and goats. The cumulative percentages of intravenous dose of kanamycin excreted through urine during 12 hours in buffaloes, cows, sheep and goats were 4.31 ± 0.37, 2.53 ± 0.30, 11.0 ± 1.04 and 15.8 ± 2.22, respectively. This species variation in the percentage of urinary excretion in these domestic ruminants coincides with their respective glomerular filtration rates, being the highest in goats, lowest in cows and intermediate in sheep and buffaloes.

  9. Microcalorimetric Evaluation of the Effect of Kanamycin: An Analysis Based on the Median-Effect Principle

    Institute of Scientific and Technical Information of China (English)

    WANG,Li-Heng; FAN,Dai-Di; SHANG,Long-An; SHI,Hui-Juan; MA,Xiao-Xuan; MI,Yu; GU,Li-Feng; XU,Kang-Zhen

    2007-01-01

    A study of the effect of drug, kanamycin, on the growth metabolism of recombinant Escherichia coli B1 was carried 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 B1 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.

  10. An Arabidopsis thaliana ABC transporter that confers kanamycin resistance in transgenic plants does not endow resistance to Escherichia coli

    OpenAIRE

    Burris, Kellie; Mentewab, Ayalew; Ripp, Steven; Stewart, C. Neal

    2007-01-01

    Summary Concerns have been raised about potential horizontal gene transfer (HGT) of antibiotic resistance markers (ARMs) from transgenic plants to bacteria of medical and environmental importance. All ARMs used in transgenic plants have been bacterial in origin, but it has been recently shown that an Arabidopsis thaliana ABC transporter, Atwbc19, confers kanamycin resistance when overexpressed in transgenic plants. Atwbc19 was evaluated for its ability to transfer kanamycin resistance to Esch...

  11. 21 CFR 524.1484d - Neomycin sulfate, hydrocortisone acetate, tetracaine hydrochloride ear ointment.

    Science.gov (United States)

    2010-04-01

    ..., tetracaine hydrochloride ear ointment. 524.1484d Section 524.1484d Food and Drugs FOOD AND DRUG..., tetracaine hydrochloride ear ointment. (a) Specifications. The product contains 5 milligrams of neomycin... a lesser degree, chronic otitis externa in dogs and cats. In treatment of ear canker and...

  12. CRAC channel is inhibited by neomycin in a Ptdlns(4,5)P2-independent manner.

    Science.gov (United States)

    Huang, Kun; Wang, Xuemei; Liu, Yanjun; Zhao, Yi

    2015-03-01

    Depletion of intracellular Ca(2+) stores evokes store-operated Ca(2+) entry through the Ca(2+) release-activated Ca(2+) (CRAC) channels. In this study, we found that the store-operated Ca(2+) entry was inhibited by neomycin, an aminoglycoside that strongly binds phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2). Patch clamp recordings revealed that neomycin blocked the CRAC currents reconstituted by co-expression of Orai1 and Stim1 in HEK293 cells. Using a rapamycin-inducible PtdIns(4,5)P2-specific phosphatase (Inp54p) system to manipulate the PtdIns(4,5)P2 in the plasma membrane, we found that the CRAC current was not altered by PtdIns(4,5)P2 depletion. This result suggests that PtdIns(4,5)P2 is not required for CRAC channel activity, and thereby, neomycin inhibits CRAC channels in a manner that is independent of neomycin-PtdIns(4,5)P2 binding.

  13. Neomycin binding preserves extracellular matrix in bioprosthetic heart valves during in vitro cyclic fatigue and storage.

    Science.gov (United States)

    Raghavan, Devanathan; Starcher, Barry C; Vyavahare, Naren R

    2009-05-01

    Bioprosthetic heart valve (BHV) cusps have a complex architecture consisting of an anisotropic arrangement of collagen, glycosaminoglycans (GAGs) and elastin. Glutaraldehyde (GLUT) is used as a fixative for all clinical BHV implants; however, it only stabilizes the collagen component of the tissue, and other components such as GAGs and elastin are lost from the tissue during processing, storage or after implantation. We have shown previously that the effectiveness of the chemical crosslinking can be increased by incorporating neomycin trisulfate, a hyaluronidase inhibitor, to prevent the enzyme-mediated GAG degradation. In the present study, we optimized carbodiimide-based GAG-targeted chemistry to incorporate neomycin into BHV cusps prior to conventional GLUT crosslinking. This crosslinking leads to enhanced preservation of GAGs during in vitro cyclic fatigue and storage. The neomycin group showed greater GAG retention after both 10 and 50 million accelerated fatigue cycles and after 1 year of storage in GLUT solution. Thus, additional binding of neomycin to the cusps prior to standard GLUT crosslinking could enhance tissue stability and thus heart valve durability.

  14. Recognition of HIV-TAR RNA using neomycin-benzimidazole conjugates.

    Science.gov (United States)

    Ranjan, Nihar; Kumar, Sunil; Watkins, Derrick; Wang, Deyun; Appella, Daniel H; Arya, Dev P

    2013-10-15

    Synthesis of a novel class of compounds and their biophysical studies with TAR-RNA are presented. The synthesis of these compounds was achieved by conjugating neomycin, an aminoglycoside, with benzimidazoles modeled from a B-DNA minor groove binder, Hoechst 33258. The neomycin-benzimidazole conjugates have varying linkers that connect the benzimidazole and neomycin units. The linkers of varying length (5-23 atoms) in these conjugates contain one to three triazole units. The UV thermal denaturation experiments showed that the conjugates resulted in greater stabilization of the TAR-RNA than either neomycin or benzimidazole used in the synthesis of conjugates. These results were corroborated by the FID displacement and tat-TAR inhibition assays. The binding of ligands to the TAR-RNA is affected by the length and composition of the linker. Our results show that increasing the number of triazole groups and the linker length in these compounds have diminishing effect on the binding to TAR-RNA. Compounds that have shorter linker length and fewer triazole units in the linker displayed increased affinity towards the TAR RNA.

  15. Growth factors have a protective effect on neomycin-induced hair cell loss.

    Science.gov (United States)

    Lou, Xiangxin; Yuan, Huihua; Xie, Jing; Wang, Xianliu; Yang, Liangliang; Zhang, Yanzhong

    2015-01-01

    We have demonstrated that selected growth factors are involved in regulating survival and proliferation of progenitor cells derived from the neonatal rat organ of Corti (OC). The protective and regenerative effects of these defined growth factors on the injured organ of Corti were therefore investigated. The organ of Corti dissected from the Wistar rat pups (P3-P5) was split into apical, middle, and basal parts, explanted and cultured with or without neomycin and growth factors. Insulin-like growth factor-1 (IGF-1), fibroblast growth factor-2 (FGF-2), and epidermal growth factor (EGF) protected the inner hair cells (IHCs) and outer hair cells (OHCs) from neomycin ototoxicity. Using EGF, IGF-1, and FGF-2 alone induced no protective effect on the survival of auditory hair cells. Combining 2 growth factors (EGF + IGF-1, EGF + FGF-2, or IGF-1 + FGF-2) gave statistically protective effects. Similarly, combining all three growth factors effectively protected auditory hair cells from the ototoxic insult. None of the growth factors induced regeneration of hair cells in the explants injured with neomycin. Thus various combinations of the three defined factors (IGF-1, FGF-2, and EGF) can protect the auditory hair cells from the neomycin-induced ototoxic damage, but no regeneration was seen. This offers a possible novel approach to the treatment of hearing loss.

  16. 21 CFR 524.960 - Flumethasone, neomycin sulfate, and polymyxin B sulfate ophthalmic solutions.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Flumethasone, neomycin sulfate, and polymyxin B sulfate ophthalmic solutions. 524.960 Section 524.960 Food and Drugs FOOD AND DRUG ADMINISTRATION... fundamental healing mechanism. Adrenocorticoid compounds have been reported to cause an increase in...

  17. 21 CFR 520.82b - Aminopropazine fumarate, neomycin sulfate tablets.

    Science.gov (United States)

    2010-04-01

    ... chapter. (c) Conditions of use. (1) The drug is used in dogs to control bacterial diarrhea caused by... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Aminopropazine fumarate, neomycin sulfate tablets. 520.82b Section 520.82b Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND...

  18. A strategy to enhance the binding affinity of fluorophore-aptamer pairs for RNA tagging with neomycin conjugation.

    Science.gov (United States)

    Jeon, Jongho; Lee, Kyung Hyun; Rao, Jianghong

    2012-10-14

    Fluorogenic sulforhodamine-neomycin conjugates have been designed and synthesized for RNA tagging. Conjugates were fluorescently activated by binding to RNA aptamers and exhibited greater than 250-400 fold enhancement in binding affinity relative to corresponding unconjugated fluorophores.

  19. Organogenesis and plant selection by kanamycin of strawberry in vitro aiming at genetic transformation

    Directory of Open Access Journals (Sweden)

    Letícia Reis

    2015-08-01

    Full Text Available The success of genetic transformation of the strawberry (Fragaria x ananassa is strongly influenced by the cultivar and depends on efficient regeneration and plant-transformed selection. In plant regeneration, efficient organogenesis depends on the concentrations of plant growth regulators that are present in the culture medium, and the response varies according to the explant type and cultivar. Similarly, in the selection of transformed plants, the sensitivity to selective agents induced by marker genes depends on the concentration of the selective substance in the regeneration medium and must be adjusted according to the chosen cultivar. The aim of this work was to test different combinations and concentrations of TDZ (Thidiazuron and IBA (indolebutyric-acid added to the base medium MS (MURASHIGE; SKOOG, 1962 for shoot regeneration of Camiño Real and Festival cultivars from leaf and petiole explants. In the case of ‘Camiño Real’, 1.0, 1.5 and 2.0 mg L-1 of TDZ combined with 0.1, 0.3 and 0.5 mg L-1 of IBA were tested. In the case of ‘Festival’, 1.0, 1.5 and 2.0 mg L-1 of TDZ combined with 0.1 and 0.3 mg L-1 of IBA were evaluated. Another aim was to define the lowest concentration of the selective agent Kanamycin (marker gene npt II able to inhibit regeneration of the untransformed shoots. For this purpose, Kanamycin doses of 0, 2, 4, 6, 8 and 10 mg L-1 were added to the pre-established regeneration medium. In the petiole explant test, normal shoot regeneration did not occur; thus, only shoot regeneration and the sensitivity to Kanamycin from leaf explants were analyzed. For ‘Camiño Real’ the best shoot regeneration was established with a combination of 2.0 mg L-1 of TDZ + 0.1 mg L-1 of IBA. On the other hand, for ‘Festival’ the best shoot regeneration was established with a combination of 2.5 mg L-1 of TDZ + 0.1 mg L-1 of IBA. As far as Kanamycin sensitivity is concerned, total inhibition of the shoots was observed with 10 mg

  20. The ryanodine receptor pore blocker neomycin also inhibits channel activity via a previously undescribed high-affinity Ca(2+) binding site.

    Science.gov (United States)

    Laver, Derek R; Hamada, Tomoyo; Fessenden, James D; Ikemoto, Noriaki

    2007-12-01

    In this study, we present evidence for the mechanism of neomycin inhibition of skeletal ryanodine receptors (RyRs). In single-channel recordings, neomycin produced monophasic inhibition of RyR open probability and biphasic inhibition of [(3)H]ryanodine binding. The half-maximal inhibitory concentration (IC(50)) for channel blockade by neomycin was dependent on membrane potential and cytoplasmic [Ca(2+)], suggesting that neomycin acts both as a pore plug and as a competitive antagonist at a cytoplasmic Ca(2+) binding site that causes allosteric inhibition. This novel Ca(2+)/neomycin binding site had a neomycin affinity of 100 nM: and a Ca(2+) affinity of 35 nM,: which is 30-fold higher than that of the well-described cytoplasmic Ca(2+) activation site. Therefore, a new high-affinity class of Ca(2+) binding site(s) on the RyR exists that mediates neomycin inhibition. Neomycin plugging of the channel pore induced brief (1-2 ms) conductance substates at 30% of the fully open conductance, whereas allosteric inhibition caused complete channel closure with durations that depended on the neomycin concentration. We quantitatively account for these results using a dual inhibition model for neomycin that incorporates voltage-dependent pore plugging and Ca(2+)-dependent allosteric inhibition.

  1. Effects of streptomycin and neomycin on the adrenal medulla of dogs.

    Science.gov (United States)

    Corrado, A P; Guimarães, A F; Prado, W A

    1984-01-01

    The blocking effects of streptomycin and neomycin upon adrenal neurotransmission were studied in dogs. The pressor responses to splanchnic nerve stimulation (SNS) or 50 micrograms/kg nicotine were both blocked by 100 mg/kg but not by 50 mg/kg of the antibiotics injected systemically through the femoral vein. The effect was reversed by intravenous infusion of calcium chloride. The pressor response to SNS, but not that to nicotine, was blocked by the antibiotics injected into the adrenal gland through the adrenolumbar vein at doses up to 10 mg/kg. This effect was reversed by 50 micrograms/kg calcium chloride. Higher doses of antibiotics blocked the response to both stimuli, with only the response to nicotine being reversed by calcium chloride. These data indicate that streptomycin and neomycin block adrenal neurotransmission by interfering with calcium ions at pre- or pre-plus postsynaptic levels depending on the dose and route of administration.

  2. DRUG NEOMYCIN RELEASE FROM CORDIA DICHOTOMA TRANSDERMAL FILM AND ANTIINFLAMMATORY ACTIVITY

    Directory of Open Access Journals (Sweden)

    A A Shahapurkar

    2011-09-01

    Full Text Available Transdermal Films were prepared using 10 % w/v natural polymer (fruit gum of Cordia dichotoma with different percentage of plasticizer (glycerin 0.10, 0.20 and 0.25 % w/v, same percentage of preservative (methyl paraben 0.1 % w/v and drug (neomycin 0.2 % w/v. The films were casted on glass plates and dried under controlled evaporation. Films prepared with 0.20 % w/v of glycerin showed satisfactory drying after 24 h. They were evaluated by the various parameters like thickness, tensile strength, water uptake, folding endurance, piercing load and skin irritation test. In the present study Cordia dichotoma Transdermal film was investigated for Anti-Inflammatory activity in carrageenan-induced rat paw edema. It was compared to control group and with that of standard drug,diclofenac sodium. The neomycin film with 0.20% plasticizer showed significant result.

  3. Neomycin fixation followed by ethanol pretreatment leads to reduced buckling and inhibition of calcification in bioprosthetic valves.

    Science.gov (United States)

    Raghavan, Devanathan; Shah, Sagar R; Vyavahare, Naren R

    2010-01-01

    Glutaraldehyde crosslinked bioprosthetic heart valves (BHVs) have two modalities of failure: degeneration (cuspal tear due to matrix failure) and calcification. They can occur independently as well as one can lead to the other causing co-existence. Calcific failure has been extensively studied before and several anti-calcification treatments have been developed; however, little research is directed to understand mechanisms of valvular degeneration. One of the shortcomings of glutaraldehyde fixation is its inability to stabilize all extracellular matrix components in the tissue. Previous studies from our lab have demonstrated that neomycin could be used as a fixative to stabilize glycosaminoglycans (GAGs) present in the valve to improve matrix properties. But neomycin fixation did not prevent cuspal calcification. In the present study, we wanted to enhance the anti-calcification potential of neomycin fixed valves by pre-treating with ethanol or removing the free aldehydes by sodium borohydride treatment. Ethanol treatment has been previously used and found to have excellent anti-calcification properties for valve cusps. Results demonstrated in this study suggest that neomycin followed by ethanol treatment effectively preserves GAGs both in vitro as well as in vivo after subdermal implantation in rats. In vivo calcification was inhibited in neomycin fixed cusps pretreated with ethanol compared to glutaraldehyde (GLUT) control. Sodium borohydride treatment by itself did not inhibit calcification nor stabilized GAGs against enzymatic degradation. Neomycin fixation followed by ethanol treatment of BHVs could prevent both modalities of failure, thereby increasing the effective durability and lifetime of these bioprostheses several fold.

  4. Inhibition of K+ currents in type I vestibular hair cells by gentamicin and neomycin.

    Science.gov (United States)

    Mann, Scott E; Johnson, Matthew; Meredith, Frances L; Rennie, Katherine J

    2013-01-01

    Significant ototoxicity limits the use of aminoglycoside (AG) antibiotics. Several mechanisms may contribute to the death of both auditory and vestibular hair cells. In this study the effects of gentamicin and neomycin on K(+) currents in mature and early postnatal type I vestibular hair cells (HCI) were tested directly. The whole-cell patch clamp technique was used to assess the effects of AG and KCNQ channel modulators on K(+) currents (IK) in HCI acutely isolated from gerbil semicircular canals. Extracellular neomycin (1 mM) rapidly reduced peak outward IK by 16 ± 4% (n = 9) in mature HCI (postnatal days, P, 25-66). Gentamicin (5 mM) reduced outward IK by 16 ± 3% (n = 8). A similar reduction in outward current was seen in immature HCI (P5-9) that lacked the low-voltage-activated component of IK observed in mature cells. Intracellular application of gentamicin and neomycin also reduced IK in mature HCI. Modulators of KCNQ channels were used to probe KCNQ channel involvement. The selective KCNQ antagonist XE991 did not reduce IK and the neomycin-induced reduction in IK was not reversed by the KCNQ agonist flupirtine. Application of intracellular poly-D-lysine to sequester PIP2 did not reduce IK. Application of the K(+) channel blocker 4-aminopyridine (4-AP) strongly reduced IK, and extracellular AG in the presence of 4-AP gave no further inhibition of IK. In summary, AG significantly reduce the 4-AP-sensitive IK in early postnatal and mature HCI. K(+) current inhibition differs from that seen in outer hair cells, since it does not appear to involve PIP2 sequestration or KCNQ channels.

  5. Transmissible Resistance to Penicillin G, Neomycin, and Chloramphenicol in Rhizobium japonicum1

    Science.gov (United States)

    Cole, Michael A.; Elkan, Gerald H.

    1973-01-01

    The genetic basis for resistance to a number of antibiotics was examined in Rhizobium japonicum. Resistance to penicillin G, neomycin, and chloramphenicol appears to be mediated by an extrachromosomal element similar to that found in the Enterobacteriaceae. Resistance to these antibiotics was eliminated from cells by treatment with acridine orange, and resistance to all three antibiotics could be transferred en bloc to Agrobacterium tumefaciens under conditions excluding transformation or transduction as possible genetic mechanisms. PMID:4491197

  6. Acute and subchronic neuromuscular blocking characteristics of streptomycin: a comparison with neomycin.

    Science.gov (United States)

    Lee, C; deSilva, A J

    1979-05-01

    The characteristics of the neuromuscular block produced by streptomycin in vivo were studied on the sciatic-tibialis anterior nerve-muscle preparation of eight anaesthetized cats. The lungs of the animals were ventilated mechanically and normocarbia was maintained. During acute exposure to streptomycin (within 2 h), ED50 for blockade of the twitch was 56 (SEM +/- 5) mg kg-1 of the base. The characteristics of block were similar to those of neomycin-induced block in some aspects. There was absence of train-of-four fade and tetanic fade, partial sparing of the responses elicited at 10 Hz and 20 Hz, and total sparing of the 50 Hz tetanus, as well as the post-tetanic twitch. In contrast to neomycin-induced neuromuscular block, however, post-tetanic exhaustion was not observed and prolonged exposure to streptomycin (22-28 h) did not change the characteristics of the block. We conclude that, despite their chemical similarities, streptomycin and neomycin block neuromuscular transmission differently.

  7. Neomycin inhibition of (+)-7-iso-jasmonoyl-L-isoleucine accumulation and signaling.

    Science.gov (United States)

    Vadassery, Jyothilakshmi; Reichelt, Michael; Jimenez-Aleman, Guillermo H; Boland, Wilhelm; Mithöfer, Axel

    2014-07-01

    The majority of plant defenses against insect herbivores are coordinated by jasmonate (jasmonic acid, JA; (+)-7-iso-jasmonoyl-L-isoleucine, JA-Ile)-dependent signaling cascades. Insect feeding and mimicking herbivory by application of oral secretions (OS) from the insect induced both cytosolic Ca(2+) and jasmonate-phytohormone elevation in plants. Here it is shown that in Arabidopsis thaliana upon treatment with OS from lepidopteran Spodoptera littoralis larvae, the antibiotic neomycin selectively blocked the accumulation of OS-induced Ca(2+) elevation and level of the bioactive JA-Ile, in contrast to JA level. Furthermore, neomycin treatment affected the downstream expression of JA-Ile-responsive genes, VSP2 and LOX2, in Arabidopsis. The neomycin-dependent reduced JA-Ile level is partially due to increased CYP94B3 expression and subsequent JA-Ile turn-over to12-hydroxy-JA-Ile. It is neither due to the inhibition of the enzymatic conjugation process nor to substrate availability. Thus, blocking Ca(2+) elevation specifically controls JA-Ile accumulation and signaling, offering an insight into role of calcium in defense against insect herbivory.

  8. Triple recognition of B-DNA by a neomycin-Hoechst 33258-pyrene conjugate.

    Science.gov (United States)

    Willis, Bert; Arya, Dev P

    2010-01-26

    Recent developments have indicated that aminoglycoside binding is not limited to RNA, but to nucleic acids that, like RNA, adopt conformations similar to its A-form. We further sought to expand the utility of aminoglycoside binding to B-DNA structures by conjugating neomycin, an aminoglycoside antibiotic, with the B-DNA minor groove binding ligand Hoechst 33258. Envisioning a dual groove binding mode, we have extended the potential recognition process to include a third, intercalative moiety. Similar conjugates, which vary in the number of binding moieties but maintain identical linkages to allow direct comparisons to be made, have also been prepared. We report herein novel neomycin- and Hoechst 33258-based conjugates developed in our laboratories for exploring the recognition potential with B-DNA. Spectroscopic studies such as UV melting, differential scanning calorimetry, isothermal fluorescence titrations, and circular dichroism together illustrate the triple recognition of the novel conjugate containing neomycin, Hoechst 33258, and pyrene. This study represents the first example of DNA molecular recognition capable of minor versus major groove recognition in conjunction with intercalation.

  9. Ion-association method for the colorimetric determination of neomycin sulphate in pure and dosage forms

    Science.gov (United States)

    Amin, A. S.; Issa, Y. M.

    2003-03-01

    A simple, fairly rapid, sensitive and accurate method is described for the colorimetric determination of neomycin sulphate (NMS), based on the measurement of the absorbance of the extracted organic soluble ion-association complex formed between neomycin dictation and a bulky counter anion. Different chromotropic acid azo dyes were examined as counter ions. The effect of pH, the counter ion concentration, sequence of addition and solvents for extraction were also illustrated. The most suitable system is based on reagent VIII (pH 7.5) with chloroform as the extraction solvent. The use of other counter ions, in conjunction with their respective solvents, was found to be less sensitive. The neomycin-reagent VIII system exhibits negligible or no interference when used for the determination of up to 58 μg ml -1 of NMS in the presence of several drug excipiences. The method has been used for the determination of up to 58 μg ml -1 with a good recovery (99.8±1.5%), and the precision is supported by the low relative standard deviation ⩽1.35%. The sensitivity is discussed and the results are compared with the official method. The proposed method was applied successfully to the determination of NMS in pure and dosage forms, with a good precision and accuracy compared to the official one.

  10. Prevalence of transposons encoding kanamycin, ampicillin and trimethoprim resistance in isolates from urinary tract infections detected using DNA probes.

    Science.gov (United States)

    Chang, S F; Chang, L L; Chow, T Y; Wu, W J; Chang, J C

    1992-03-01

    Drug resistant Gram-negative bacteria causing urinary tract infections were collected. Kanamycin, ampicillin or trimethoprim-resistant strains were analyzed separately for the presence of Tn5, Tn3, or Tn7 by colony hybridization. Of these isolates, kanamycin-resistant transposons were present in 38.2% of 60 kanamycin-resistant isolates. A 3.3 kb fragment containing SacI-BamHI transposase of Tn3 and 42.6% showed a positive reaction in 129 ampicillin-resistant clinical isolates. Among the 75 trimethoprim-resistant isolates studied, 52% were shown to contain Tn7 when probed with a 1 kb BamHI fragment of Tn7. Results from Southern hybridizations demonstrated that these antibiotic resistant genes had been born on plasmids in some clinical isolates.

  11. Exogenous alanine and/or glucose plus kanamycin kills antibiotic-resistant bacteria.

    Science.gov (United States)

    Peng, Bo; Su, Yu-Bin; Li, Hui; Han, Yi; Guo, Chang; Tian, Yao-Mei; Peng, Xuan-Xian

    2015-02-03

    Multidrug-resistant bacteria are an increasingly serious threat to human and animal health. However, novel drugs that can manage infections by multidrug-resistant bacteria have proved elusive. Here we show that glucose and alanine abundances are greatly suppressed in kanamycin-resistant Edwardsiella tarda by GC-MS-based metabolomics. Exogenous alanine or glucose restores susceptibility of multidrug-resistant E. tarda to killing by kanamycin, demonstrating an approach to killing multidrug-resistant bacteria. The mechanism underlying this approach is that exogenous glucose or alanine promotes the TCA cycle by substrate activation, which in turn increases production of NADH and proton motive force and stimulates uptake of antibiotic. Similar results are obtained with other Gram-negative bacteria (Vibrio parahaemolyticus, Klebsiella pneumoniae, Pseudomonas aeruginosa) and Gram-positive bacterium (Staphylococcus aureus), and the results are also reproduced in a mouse model for urinary tract infection. This study establishes a functional metabolomics-based strategy to manage infection by antibiotic-resistant bacteria. Copyright © 2015 Elsevier Inc. All rights reserved.

  12. 2,3-Dihydroxybenzoic acid attenuates kanamycin-induced volume reduction in mouse utricular type I hair cells

    DEFF Research Database (Denmark)

    Severinsen, Stig Åvall; Kirkegaard, Mette; Nyengaard, Jens Randel

    2006-01-01

    injection. Total volume of the utricle, as well as total number of hair and supporting cells, were estimated on light microscopic sections. Total volume and mean volume of hair cell types I and II and supporting cells were estimated on digital transmission electron micrographs. Total volume of the utricular...... macula, hair cell type I and supporting cells decreased significantly in animals injected with kanamycin but not in animals co-treated with DHB. Hair and supporting cell numbers remained unchanged in all three groups. In conclusion, the kanamycin-induced volume reduction of type I hair cells...

  13. Osteoblasts detect pericellular calcium concentration increase via neomycin-sensitive voltage gated calcium channels.

    Science.gov (United States)

    Sun, Xuanhao; Kishore, Vipuil; Fites, Kateri; Akkus, Ozan

    2012-11-01

    The mechanisms underlying the detection of critically loaded or micro-damaged regions of bone by bone cells are still a matter of debate. Our previous studies showed that calcium efflux originates from pre-failure regions of bone matrix and MC3T3-E1 osteoblasts respond to such efflux by an increase in the intracellular calcium concentration. The mechanisms by which the intracellular calcium concentration increases in response to an increase in the pericellular calcium concentration are unknown. Elevation of the intracellular calcium may occur via release from the internal calcium stores of the cell and/or via the membrane bound channels. The current study applied a wide range of pharmaceutical inhibitors to identify the calcium entry pathways involved in the process: internal calcium release from endoplasmic reticulum (ER, inhibited by thapsigargin and TMB-8), calcium receptor (CaSR, inhibited by calhex), stretch-activated calcium channel (SACC, inhibited by gadolinium), voltage-gated calcium channels (VGCC, inhibited by nifedipine, verapamil, neomycin, and ω-conotoxin), and calcium-induced-calcium-release channel (CICRC, inhibited by ryanodine and dantrolene). These inhibitors were screened for their effectiveness to block intracellular calcium increase by using a concentration gradient induced calcium efflux model which mimics calcium diffusion from the basal aspect of cells. The inhibitor(s) which reduced the intracellular calcium response was further tested on osteoblasts seeded on mechanically loaded notched cortical bone wafers undergoing damage. The results showed that only neomycin reduced the intracellular calcium response in osteoblasts, by 27%, upon extracellular calcium stimulus induced by concentration gradient. The inhibitory effect of neomycin was more pronounced (75% reduction in maximum fluorescence) for osteoblasts seeded on notched cortical bone wafers loaded mechanically to damaging load levels. These results imply that the increase in

  14. Glutamate co-transmission from developing medial nucleus of the trapezoid body - Lateral superior olive synapses is cochlear dependent in kanamycin-treated rats

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Jae Ho [Institute of Tissue Regeneration Engineering (ITREN), Dankook University, San 29, Anseo-dong, Cheonan-si, Chungnam 330-714 (Korea, Republic of); Pradhan, Jonu [Department of Nanobio Medical Science, Dankook University, San 29, Anseo-dong, Cheonan-si, Chungnam 330-714 (Korea, Republic of); Maskey, Dhiraj; Park, Ki Sup [Department of Anatomy, College of Medicine, Dankook University, San 29, Anseo-dong, Cheonan-si, Chungnam 330-714 (Korea, Republic of); Hong, Sung Hwa [Department of Otorhinolaryngology-Head and Neck Surgery, Samsung Medical Center, Sungkyunkwan University, School of Medicine, 50, Irwon-dong, Gangnam-gu, Seoul 135-710 (Korea, Republic of); Suh, Myung-Whan [Department of Otorhinolaryngology-Head and Neck Surgery, College of Medicine, Dankook University, San 29, Anseo-dong, Cheonan-si, Chungnam 330-714 (Korea, Republic of); Kim, Myeung Ju, E-mail: mjukim99@dankook.ac.kr [Department of Anatomy, College of Medicine, Dankook University, San 29, Anseo-dong, Cheonan-si, Chungnam 330-714 (Korea, Republic of); Ahn, Seung Cheol, E-mail: ansil67@hanmail.net [Department of Physiology, College of Medicine, Dankook University, San 29, Anseo-dong, Cheonan-si, Chungnam 330-714 (Korea, Republic of)

    2011-02-11

    Research highlights: {yields} Glutamate co-transmission is enhanced in kanamycin-treated rats. {yields} VGLUT3 expression is increased in kanamycin-treated rats. {yields} GlyR expression is decreased in kanamycin-treated rats. {yields} GlyR, VGLUT3 expression patterns are asymmetric in unilaterally cochlear ablated rat. -- Abstract: Cochlear dependency of glutamate co-transmission at the medial nucleus of the trapezoid body (MNTB) - the lateral superior olive (LSO) synapses was investigated using developing rats treated with high dose kanamycin. Rats were treated with kanamycin from postnatal day (P) 3 to P8. A scanning electron microscopic study on P9 demonstrated partial cochlear hair cell damage. A whole cell voltage clamp experiment demonstrated the increased glutamatergic portion of postsynaptic currents (PSCs) elicited by MNTB stimulation in P9-P11 kanamycin-treated rats. The enhanced VGLUT3 immunoreactivities (IRs) in kanamycin-treated rats and asymmetric VGLUT3 IRs in the LSO of unilaterally cochlear ablated rats supported the electrophysiologic data. Taken together, it is concluded that glutamate co-transmission is cochlear-dependent and enhanced glutamate co-transmission in kanamycin-treated rats is induced by partial cochlear damage.

  15. Determination of neomycin sulfate and impurities using high-performance anion-exchange chromatography with integrated pulsed amperometric detection.

    Science.gov (United States)

    Hanko, Valoran P; Rohrer, Jeffrey S

    2007-01-01

    Neomycin B is one of a class of aminoglycoside antibiotics that lack a good chromophore, and is therefore difficult to determine using reversed-phase HPLC with absorbance detection. This is especially true for determining the quantity of each impurity. We show that neomycin sulfate and its major impurities, including neamine (neomycin A), can be separated on a strong anion-exchange column using a weak potassium hydroxide eluent (2.40 mM) at a column temperature of 30 degrees C, and directly detected by integrated pulsed amperometric detection (IPAD). The resolution (United States Pharmacopeia (USP) definition) between neomycin B and the closest major impurity ranged from 6.56 and 7.45 over 10 days of consecutive analysis (7.24+/-0.10, n=836 injections). Due to the difficulty of producing weak hydroxide eluents of the required purity (i.e. carbonate-free), this method depends on automatic eluent generation to ensure method ruggedness. This method exhibited good long-term (10 days, 822 injections) retention time stability with a R.S.D. of 0.6%. Peak area R.S.D. (10 microM) was 1.3%. Method robustness was evaluated by intentionally varying the flow rate, eluent concentration, column temperature, and column. The spike recoveries of neomycin B from extractions of three different topical ointments and cream formulations ranged from 95 to 100%. The measured concentration of neomycin B in these formulations ranged from 119 to 154% of the label concentration. The R.S.D. for the measured concentration of one of the formulations tested over three separate days, n=11 extracts, was 3.2%. Based on the results of these evaluations, we believe this method can be used for neomycin sulfate identity, assay, and purity.

  16. Limited sampling strategies for therapeutic drug monitoring of amikacin and kanamycin in patients with multidrug-resistant tuberculosis

    NARCIS (Netherlands)

    Dijkstra, J.A.; van Altena, R.; Akkerman, O. W.; de Lange, W. C. M.; Proost, J. H.; van der Werf, T. S.; Kosterink, J. G. W.; Alffenaar, J. W. C.

    2015-01-01

    Amikacin and kanamycin are considered important and effective drugs in the treatment of multidrug-resistant tuberculosis (MDR-TB). Unfortunately, the incidence of toxicity is high and is related to elevated drug exposure. In order to achieve a balance between efficacy and toxicity, a population phar

  17. Mobilization properties of small ColE1-like plasmids carrying kanamycin resistance gene isolated from Salmonella enterica serotypes

    Science.gov (United States)

    Background: Previously we isolated and characterized various groups of small kanamycin resistance (KanR) ColE1-like plasmids from different serotypes of Salmonella enterica isolates. These plasmids all carried the aph(3)-I gene encoding the aminoglycoside phosphotransferase responsible for the kanam...

  18. Pseudothrombocytopenia observed with ethylene diamine tetra acetate and citrate anticoagulants, resolved using 37°C incubation and Kanamycin

    Directory of Open Access Journals (Sweden)

    Vandana Kamath

    2013-01-01

    Full Text Available Pseudothrombocytopenia (PTP is defined by falsely low platelet counts on automated analyzers caused by in vitro phenomena including large platelet aggregates in blood samples. Diagnosis and resolution of PTP is crucial as it can lead to unwarranted interventions. We discuss a case of PTP in a pre-surgical setting, which was resolved using 37°C incubation and Kanamycin.

  19. Pseudothrombocytopenia observed with ethylene diamine tetra acetate and citrate anticoagulants, resolved using 37°C incubation and Kanamycin.

    Science.gov (United States)

    Kamath, Vandana; Sarda, Parimal; Chacko, Mary Purna; Sitaram, Usha

    2013-01-01

    Pseudothrombocytopenia (PTP) is defined by falsely low platelet counts on automated analyzers caused by in vitro phenomena including large platelet aggregates in blood samples. Diagnosis and resolution of PTP is crucial as it can lead to unwarranted interventions. We discuss a case of PTP in a pre-surgical setting, which was resolved using 37°C incubation and Kanamycin.

  20. In vitro evaluation of the synergistic activity of neomycin-polymyxin B association against pathogens responsible for otitis externa.

    Science.gov (United States)

    Tempera, G; Mangiafico, A; Genovese, C; Giudice, E; Mastrojeni, S; Nicolosi, D; Ferneri, P M

    2009-01-01

    The most recent guidelines recommend, for otitis externa antibiotic therapy, the use of topical formulations in that they are very safe, have a quicker effect and do not induce bacterial resistance compared to systemic therapy. The choice of the class of antibiotics in empiric therapy of otitis externa must take into consideration the polymicrobic nature of the infection that includes both bacteria (Grampositive and Gram-negative) and mycetes. For this reason, in this study we evaluated the synergic activity of neomycin in association with polymyxin B against the pathogens commonly responsible for otitis externa, compared to that of a single antibiotic (ciprofloxacin). The polymyxinB/neomycin association shows clear synergic effects with values of both Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) reduced by 3-4 times with respect to the single antibiotic; and in P. aeruginosa the synergistic effect of the neomycin/polymyxin B association with respect to neomycin was more evident (5-6 times), with an intrinsic in vitro activity constantly higher than that of ciprofloxacin alone or in association with hydrocortisone. From the analysis of the data obtained in vitro, we can conclude that the possibility of using a topical formulation containing a synergistic association of antibiotics, such as neomycin-polymyxin B, in such a way as to obtain the maximum effect in the minimum time with an increase in the spectrum of action of non-bacterial pathogens, is an optimal choice for the clinician for the empiric therapy of otitis externa.

  1. Effects of the aminoglycoside antibiotics, streptomycin and neomycin, on neuromuscular transmission. I. Presynaptic considerations.

    Science.gov (United States)

    Fiekers, J F

    1983-06-01

    The effects of two aminoglycoside antibiotics, streptomycin and neomycin, were studied in voltage-clamped transected twitch fibers of the costocutaneous muscles of garter snakes (species Thamnophis). The concentration-dependent effects of each antibiotic were quantitated by measuring miniature end-plate currents (mepcs) and evoked end-plate currents (epcs) in a single fiber before and in the presence of a wide range of concentrations of each antibiotic. The amplitude and the kinetics of these currents were studied and estimates of the quantal content of evoked transmitter release determined by the direct method of mean ratios, epc/mepc. A distinct separation was obtained between the concentrations of each antibiotic which demonstrated either pre- or postsynaptic actions. Both streptomycin and neomycin produced a concentration-dependent reduction in epc amplitude at concentrations which did not reduce mepc amplitude. Thus, the primary site of action for these antibiotics was considered of presynaptic origin. Streptomycin was approximately one-tenth as active as neomycin in reducing quantal release of acetylcholine. The marked depression in epc amplitude and quantal content produced by high concentrations of each antibiotic were reversed by elevating the external calcium concentration. Double logarithmic plots of the relationship between external calcium concentration and epc amplitude yielded a slope of approximately 3.8 in control physiological solution. In the presence of blocking concentrations of each antibiotic, increasing the external calcium concentration caused a parallel shift to the right of this relationship. These results suggest that the major mechanism for the neuromuscular depression produced by these aminoglycoside antibiotics is a competitive antagonism with calcium for a common presynaptic site required for evoked transmitter release.

  2. Synthesis and spectroscopic studies of the aminoglycoside (neomycin)--perylene conjugate binding to human telomeric DNA.

    Science.gov (United States)

    Xue, Liang; Ranjan, Nihar; Arya, Dev P

    2011-04-12

    Synthesis of a novel perylene-neomycin conjugate (3) and the properties of its binding to human telomeric G-quadruplex DNA, 5'-d[AG3(T2AG3)3] (4), are reported. Various spectroscopic techniques were employed to characterize the binding of conjugate 3 to 4. A competition dialysis assay revealed that 3 preferentially binds to 4, in the presence of other nucleic acids, including DNA, RNA, DNA-RNA hybrids, and other higher-order structures (single strands, duplexes, triplexes, other G-quadruplexes, and the i-motif). UV thermal denaturation studies showed that thermal stabilization of 4 increases as a function of the increasing concentration of 3. The fluorescence intercalator displacement (FID) assay displayed a significantly tighter binding of 3 with 4 as compared to its parent constituents [220-fold stronger than neomycin (1) and 4.5-fold stronger than perylene diamine (2), respectively]. The binding of 3 with 4 resulted in pronounced changes in the molar ellipticity of the DNA absorption region as confirmed by circular dichroism. The UV-vis absorption studies of the binding of 3 to 4 resulted in a red shift in the spectrum of 3 as well as a marked hypochromic change in the perylene absorption region, suggesting that the ligand-quadruplex interaction involves stacking of the perylene moiety. Docking studies suggest that the perylene moiety serves as a bridge that end stacks on 4, making contacts with two thymine bases in the loop, while the two neomycin moieties branch into the grooves of 4.

  3. Complete genome sequence of Marivirga tractuosa type strain (H-43T)

    Energy Technology Data Exchange (ETDEWEB)

    Pagani, Ioanna [Joint Genome Institute, Walnut Creek, California; Chertkov, Olga [Los Alamos National Laboratory (LANL); Lapidus, Alla L. [Joint Genome Institute, Walnut Creek, California; Lucas, Susan [Joint Genome Institute, Walnut Creek, California; Glavina Del Rio, Tijana [Joint Genome Institute, Walnut Creek, California; Tice, Hope [Joint Genome Institute, Walnut Creek, California; Copeland, A [U.S. Department of Energy, Joint Genome Institute; Cheng, Jan-Fang [Joint Genome Institute, Walnut Creek, California; Nolan, Matt [Joint Genome Institute, Walnut Creek, California; Saunders, Elizabeth H [Los Alamos National Laboratory (LANL); Pitluck, Sam [Joint Genome Institute, Walnut Creek, California; Held, Brittany [Los Alamos National Laboratory (LANL); Goodwin, Lynne A. [Los Alamos National Laboratory (LANL); Liolios, Konstantinos [Joint Genome Institute, Walnut Creek, California; Ovchinnikova, Galina [U.S. Department of Energy, Joint Genome Institute; Ivanova, N [U.S. Department of Energy, Joint Genome Institute; Mavromatis, K [U.S. Department of Energy, Joint Genome Institute; Pati, Amrita [U.S. Department of Energy, Joint Genome Institute; Chen, Amy [Joint Genome Institute, Walnut Creek, California; Palaniappan, Krishna [Joint Genome Institute, Walnut Creek, California; Land, Miriam L [ORNL; Hauser, Loren John [ORNL; Jeffries, Cynthia [Oak Ridge National Laboratory (ORNL); Detter, J. Chris [Joint Genome Institute, Walnut Creek, California; Han, Cliff [Los Alamos National Laboratory (LANL); Tapia, Roxanne [Los Alamos National Laboratory (LANL); Ngatchou, Olivier Duplex [HZI - Helmholtz Centre for Infection Research, Braunschweig, Germany; Rohde, Manfred [HZI - Helmholtz Centre for Infection Research, Braunschweig, Germany; Goker, Markus [DSMZ - German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany; Spring, Stefan [DSMZ - German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany; Sikorski, Johannes [DSMZ - German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany; Woyke, Tanja [Joint Genome Institute, Walnut Creek, California; Bristow, James [Joint Genome Institute, Walnut Creek, California; Eisen, Jonathan [Joint Genome Institute, Walnut Creek, California; Markowitz, Victor [Joint Genome Institute, Walnut Creek, California; Hugenholtz, Philip [U.S. Department of Energy, Joint Genome Institute; Klenk, Hans-Peter [DSMZ - German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany; Kyrpides, Nikos C [Joint Genome Institute, Walnut Creek, California

    2011-01-01

    Marivirga tractuosa (Lewin 1969) Nedashkovskaya et al. 2010 is the type species of the genus Marivirga, which belongs to the family Flammeovirgaceae. Members of this genus are of interest because of their gliding motility. The species is of interest because representative strains show resistance to several antibiotics, including gentamicin, kanamycin, neomycin, polymixin and streptomycin. This is the first complete genome sequence of a member of the family Flammeovirgaceae. Here we describe the features of this organism, together with the complete genome sequence and annotation. The 4,511,574 bp long chromosome and the 4,916 bp plasmid with their 3,808 protein-coding and 49 RNA genes are a part of the Genomic Encyclopedia of Bacteria and Archaea project.

  4. Influence of subtherapeutic levels of a combination of neomycin and oxytetracycline on Salmonella typhimurium in swine, calves, and chickens.

    Science.gov (United States)

    Girard, A E; English, A R; Evangelisti, D G; Lynch, J E; Solomons, I A

    1976-07-01

    Subtherapeutic levels of oxytetracycline plus neomycin in animal feeds did not bring about increases in the quantity, prevalence, or shedding of Salmonella typhimurium in swine, calves, or chickens. In fact, the medication generally reduced the proportion of animals carrying S. typhimurium. The medicated groups were fed rations containing oxytetracycline plus neomycin commencing 5 days prior to oral inoculation with S. typhimurium and continuing through a 28-day postinoculation period. Colonization of S. typhimurium occurred in all three animal species, as evidenced by clinical signs of infection and/or colony counts in feces. Only from swine and on only one occasion was a single resistant colony isolated. It is concluded that no evidence has been obtained which would implicate the continuous low-level feeding of oxytetracycline and neomycin for a 4-week period to a potential increased incidence of disease in animals or as a hazard to humans.

  5. Inhibition of the activation and recruitment of microglia-like cells protects against neomycin-induced ototoxicity.

    Science.gov (United States)

    Sun, Shan; Yu, Huiqian; Yu, Hui; Honglin, Mei; Ni, Wenli; Zhang, Yanping; Guo, Luo; He, Yingzi; Xue, Zhen; Ni, Yusu; Li, Jin; Feng, Yi; Chen, Yan; Shao, Ruijin; Chai, Renjie; Li, Huawei

    2015-02-01

    One of the most unfortunate side effects of aminoglycoside (AG) antibiotics such as neomycin is that they target sensory hair cells (HCs) and can cause permanent hearing impairment. We have observed HC loss and microglia-like cell (MLC) activation in the inner ear (cochlea) following neomycin administration. We focused on CX3CL1, a membrane-bound glycoprotein expressed on neurons and endothelial cells, as a way to understand how the MLCs are activated and the role these cells play in HC loss. CX3CL1 is the exclusive ligand for CX3CR1, which is a chemokine receptor expressed on the surface of macrophages and MLCs. In vitro experiments showed that the expression levels of CX3CL1 and CX3CR1 increased in the cochlea upon neomycin treatment, and CX3CL1 was expressed on HCs, while CX3CR1 was expressed on MLCs. When cultured with 1 μg/mL exogenous CX3CL1, MLCs were activated by CX3CL1, and the cytokine level was increased in the cochleae leading to apoptosis in the HCs. In CX3CR1 knockout mice, a significantly greater number of cochlear HCs survived than in wild-type mice when the cochlear explants were cultured with neomycin in vitro. Furthermore, inhibiting the activation of MLCs with minocycline reduced the neomycin-induced HC loss and improved the hearing function in neomycin-treated mice in vivo. Our results demonstrate that CX3CL1-induced MLC activation plays an important role in the induction of HC death and provide evidence for CX3CL1 and CX3CR1 as promising new therapeutic targets for the prevention of hearing loss.

  6. Sodium Selenite Acts as an Otoprotectant against Neomycin-Induced Hair Cell Damage in a Zebrafish Model.

    Science.gov (United States)

    Chang, Jiwon; Choi, June; Rah, Yoon Chan; Yoo, Myung Hoon; Oh, Kyoung Ho; Im, Gi Jung; Lee, Seung Hoon; Kwon, Soon Young; Park, Hae-Chul; Chae, Sung Won; Jung, Hak Hyun

    2016-01-01

    Sodium selenite is a trace element essential for many physiological functions in the body. It is involved in various biological processes; it acts as a cofactor for antioxidant enzymes that protect against free radicals and is reported to limit metal-mediated oxidative DNA damage. In the present study, we investigated the effect of sodium selenite on neomycin ototoxicity in wild-type and transgenic zebrafish (Brn3C: EGFP). Five or six days post-fertilization, zebrafish larvae were co-exposed to 125 μM neomycin and various concentrations (10 μM, 100 μM, 250 μM, and 500 μM) of sodium selenite for 1 h. Hair cells within neuromasts of the supraorbital (SO1 and SO2), otic (O1), and occipital (OC1) lateral lines were analyzed by fluorescence microscopy (n = 10 fish per treatment). Hair cell survival was estimated as the ratio of the hair cell numbers in each group compared to those of the control group that were not exposed to neomycin. Apoptosis and hair cell damage of neuromasts were evaluated using the terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick end labeling (TUNEL) assay and 2-[4-(dimethylamino) styryl]-N-ethylpyridinium iodide (DASPEI) assay, respectively. Ultrastructural changes were evaluated using scanning electron microscopy and transmission electron microscopy. Neuromast hair cells were preserved in zebrafish exposed to 125 μM neomycin and 500 μM sodium selenite for 1 h. Sodium selenite protected against neomycin-induced hair cell loss of neuromasts, reduced apoptosis, and prevented zebrafish ultrastructural changes. We propose that sodium selenite protects against neomycin-induced hair cell damage by inhibiting apoptosis, decreasing the disarray of stereocilia, and preventing ultrastructural changes in the neuromast hair cells of the zebrafish.

  7. Development and characterization of a transdermal patch and an emulgel containing kanamycin intended to be used in the treatment of mycetoma caused by Actinomadura madurae.

    Science.gov (United States)

    López-Cervantes, Miriam; Escobar-Chávez, José Juan; Casas-Alancaster, Norma; Quintanar-Guerrero, David; Ganem-Quintanar, Adriana

    2009-12-01

    Mycetoma is a chronic, degenerative, and incapacitating infection of the skin and subcutaneous tissue. This study focuses on developing a kanamycin-based auxiliary system intended to be used in the treatment of mycetoma caused by Actinomadura madurae. Transdermal patches (with two different formulations: one with free kanamycin [K] and the other one with kanamycin adsorbed in silica [K-SG]) and an emulgel were developed. Both patches were prepared by the casting-evaporation technique. To characterize them, differential scanning calorimetry, bioadhesion, post-moisture detachment, strength and rupture distance, gas exchange, water uptake, and dissolution studies were carried out. The emulgel (containing 0.57% of kanamycin) was prepared from an oil-in-water emulsion, which was then incorporated to a gel. the patches with the best characteristics contained 22.9% of silica and 14.6% of kanamycin. Dissolution studies indicated that 8.8% of kanamycin released from K and 3.2% from K-SG at 24h. The emulgel containing 0.57% of kanamycin showed good technological characteristics for its application to the skin (viscosity, 44.9 +/- 1.4 poises; pH, 6.9 +/- 0.4; and penetrability, 52.7 +/- 5.1). The optimal patches were those containing 15.9% of freely dispersed kanamycin (K) and 14.6% of kanamycin adsorbed in silica (K-SG), which corresponds to the batch 2-0.8. The assessments performed to both pharmaceutical forms (patches and emulgel) show that they have the adequate technological characteristics for being used as an auxiliary in the treatment of actinomycetoma caused by A. madurae.

  8. The effects of neomycin and oxytetracycline alone or combined upon the incidence of salmonellosis in broiler chickens.

    Science.gov (United States)

    Williams, B J

    1985-08-01

    Chickens were orally inoculated with Salmonella typhimurium and fed rations medicated with either 200 g/ton neomycin sulfate, 200 g/ton oxytetracycline, or a combination of 200 g/ton neomycin sulfate plus 200 g/ton oxytetracycline for 16 days. The incidence of salmonellosis was lower in chickens fed the combined antibiotics, and the numbers of viable S. typhimurium in feces were significantly fewer than in chickens receiving only one antibiotic. Chickens fed the combination also gained significantly more weight on less feed than those fed only one antibiotic.

  9. Determination of neomycin and related substances in pharmaceutical preparations by reversed-phase high performance liquid chromatography with mass spectrometry and charged aerosol detection.

    Science.gov (United States)

    Stypulkowska, K; Blazewicz, A; Fijalek, Z; Warowna-Grzeskiewicz, M; Srebrzynska, K

    2013-03-25

    A new, simple and repeatable liquid chromatographic method with charged aerosol detection (LC-CAD) for determination of neomycin and related substances has been developed. Analysis of neomycin or other aminoglycosides is problematic due to a lack of chromophores. Universal response of CAD enables direct quantification of neomycin and related substances, for which no reference standard are available. Separation was performed on C18 Hypersil(®) Gold aQ column using water, methanol and heptaflurobutyric acid as mobile phase. Under developed chromatographic conditions all impurities were well separated from neomycin B. Peaks identification was evaluated by electrospray ionization mass spectrometry. The proposed method was validated according to ICH guidelines and applied to the content determination of neomycin and related substances in pharmaceutical preparations.

  10. Characterization of RbmD (glycosyltransferase in ribostamycin gene cluster) through neomycin production reconstituted from the engineered Streptomyces fradiae BS1.

    Science.gov (United States)

    Nepal, Keshav Kumar; Oh, Tae-Jin; Subba, Bimala; Yoo, Jin Cheol; Sohng, Jae Kyung

    2009-01-31

    Amino acid homology analysis predicted that rbmD, a putative glycosyltransferase from Streptomyces ribosidificus ATCC 21294, has the highest homology with neoD in neomycin biosynthesis. S. fradiae BS1, in which the production of neomycin was abolished, was generated by disruption of the neoD gene in the neomycin producer S. fradiae. The restoration of neomycin by self complementation suggested that there was no polar effect in the mutant. In addition, S. fradiae BS6 was created with complementation by rbmD in S. fradiae BS1, and secondary metabolite analysis by ESI/MS, LC/MS and MS/MS showed the restoration of neomycin production in S. fradiae BS6. These gene inactivation and complementation studies suggested that, like neoD, rbmD functions as a 2-N-acetlyglucosaminyltransferase and demonstrated the potential for the generation of novel aminoglycoside antibiotics using glycosyltransferases in vivo.

  11. Growth of soil bacteria, on penicillin and neomycin, not previously exposed to these antibiotics.

    Science.gov (United States)

    Zhang, Qichun; Dick, Warren A

    2014-09-15

    There is growing evidence that bacteria, in the natural environment (e.g. the soil), can exhibit naturally occurring resistance/degradation against synthetic antibiotics. Our aim was to assess whether soils, not previously exposed to synthetic antibiotics, contained bacterial strains that were not only antibiotic resistant, but could actually utilize the antibiotics for energy and nutrients. We isolated 19 bacteria from four diverse soils that had the capability of growing on penicillin and neomycin as sole carbon sources up to concentrations of 1000 mg L(-1). The 19 bacterial isolates represent a diverse set of species in the phyla Proteobacteria (84%) and Bacteroidetes (16%). Nine antibiotic resistant genes were detected in the four soils but some of these genes (i.e. tetM, ermB, and sulI) were not detected in the soil isolates indicating the presence of unculturable antibiotic resistant bacteria. Most isolates that could subsist on penicillin or neomycin as sole carbon sources were also resistant to the presence of these two antibiotics and six other antibiotics at concentrations of either 20 or 1000 mg L(-1). The potentially large and diverse pool of antibiotic resistant and degradation genes implies ecological and health impacts yet to be explored and fully understood.

  12. Neomycin enhances extracellular matrix stability of glutaraldehyde crosslinked bioprosthetic heart valves.

    Science.gov (United States)

    Friebe, Vincent M; Mikulis, Brandon; Kole, Sourav; Ruffing, Christy S; Sacks, Michael S; Vyavahare, Naren R

    2011-11-01

    Glutaraldehyde (GLUT) crosslinked porcine aortic heart valves are continued to be extensively used in heart valve replacement surgeries. GLUT does not crosslink glycosaminoglycans in the tissue and we have demonstrated that GAG loss is associated with tissue degeneration. In this study, we examined the ability of neomycin to enhance GLUT crosslinking to stabilize GAGs, as well as provide evidence of improved functional integrity. Neomycin enhanced GLUT crosslinked (NG) leaflets exposed to collagenase and elastase enzymes exhibited an increased resistance to proteolytic degradation. Furthermore, NG leaflets exhibited small but significant increases in collagen denaturation temperatures when compared to that of standard GLUT crosslinked BHVs. NG leaflets subjected to storage, accelerated cyclic fatigue, and in vitro enzyme mediated GAG degradation revealed improved GAG stabilization versus standard GLUT crosslinked valves, which sustained substantial decreases in GAG content. Ultrastructural analysis using transmission electron microscopy qualitatively confirmed NG leaflets preserved GAGs after enzymatic degradation. Biomechanical analyses demonstrated that NG leaflets were functionally similar to GLUT tissues but were slightly stiffer under both planar biaxial tension and under flexure. Interestingly, after GAGase treatment, GLUT tissues showed increased areal compliance and reduced hysteresis, while NG leaflets were unchanged. Collectively, NG cross-linking functionally insulated the tissue from GAG digestion, and imparted modest additional matrix stiffness but maintained tissue hysteresis properties.

  13. Identification of nuclear phosphatidylinositol 4,5-bisphosphate-interacting proteins by neomycin extraction.

    Science.gov (United States)

    Lewis, Aurélia E; Sommer, Lilly; Arntzen, Magnus Ø; Strahm, Yvan; Morrice, Nicholas A; Divecha, Nullin; D'Santos, Clive S

    2011-02-01

    Considerable insight into phosphoinositide-regulated cytoplasmic functions has been gained by identifying phosphoinositide-effector proteins. Phosphoinositide-regulated nuclear functions however are fewer and less clear. To address this, we established a proteomic method based on neomycin extraction of intact nuclei to enrich for nuclear phosphoinositide-effector proteins. We identified 168 proteins harboring phosphoinositide-binding domains. Although the vast majority of these contained lysine/arginine-rich patches with the following motif, K/R-(X(n= 3-7)-K-X-K/R-K/R, we also identified a smaller subset of known phosphoinositide-binding proteins containing pleckstrin homology or plant homeodomain modules. Proteins with no prior history of phosphoinositide interaction were identified, some of which have functional roles in RNA splicing and processing and chromatin assembly. The remaining proteins represent potentially other novel nuclear phosphoinositide-effector proteins and as such strengthen our appreciation of phosphoinositide-regulated nuclear functions. DNA topology was exemplar among these: Biochemical assays validated our proteomic data supporting a direct interaction between phosphatidylinositol 4,5-bisphosphate and DNA Topoisomerase IIα. In addition, a subset of neomycin extracted proteins were further validated as phosphatidyl 4,5-bisphosphate-interacting proteins by quantitative lipid pull downs. In summary, data sets such as this serve as a resource for a global view of phosphoinositide-regulated nuclear functions.

  14. A novel inducible protein production system and neomycin resistance as selection marker for Methanosarcina mazei.

    Science.gov (United States)

    Mondorf, Sebastian; Deppenmeier, Uwe; Welte, Cornelia

    2012-01-01

    Methanosarcina mazei is one of the model organisms for the methanogenic order Methanosarcinales whose metabolism has been studied in detail. However, the genetic toolbox is still limited. This study was aimed at widening the scope of utilizable methods in this group of organisms. (i) Proteins specific to methanogens are oftentimes difficult to produce in E. coli. However, a protein production system is not available for methanogens. Here we present an inducible system to produce Strep-tagged proteins in Ms. mazei. The promoter p1687, which directs the transcription of methyl transferases that demethylate methylamines, was cloned into plasmid pWM321 and its activity was determined by monitoring β-glucuronidase production. The promoter was inactive during growth on methanol but was rapidly activated when trimethylamine was added to the medium. The gene encoding the β-glucuronidase from E. coli was fused to a Strep-tag and was cloned downstream of the p1687 promoter. The protein was overproduced in Ms. mazei and was purified in an active form by affinity chromatography. (ii) Puromycin is currently the only antibiotic used as a selectable marker in Ms. mazei and its relatives. We established neomycin resistance as a second selectable marker by designing a plasmid that confers neomycin resistance in Ms. mazei.

  15. A Novel Inducible Protein Production System and Neomycin Resistance as Selection Marker for Methanosarcina mazei

    Directory of Open Access Journals (Sweden)

    Sebastian Mondorf

    2012-01-01

    Full Text Available Methanosarcina mazei is one of the model organisms for the methanogenic order Methanosarcinales whose metabolism has been studied in detail. However, the genetic toolbox is still limited. This study was aimed at widening the scope of utilizable methods in this group of organisms. (i Proteins specific to methanogens are oftentimes difficult to produce in E. coli. However, a protein production system is not available for methanogens. Here we present an inducible system to produce Strep-tagged proteins in Ms. mazei. The promoter p1687, which directs the transcription of methyl transferases that demethylate methylamines, was cloned into plasmid pWM321 and its activity was determined by monitoring β-glucuronidase production. The promoter was inactive during growth on methanol but was rapidly activated when trimethylamine was added to the medium. The gene encoding the β-glucuronidase from E. coli was fused to a Strep-tag and was cloned downstream of the p1687 promoter. The protein was overproduced in Ms. mazei and was purified in an active form by affinity chromatography. (ii Puromycin is currently the only antibiotic used as a selectable marker in Ms. mazei and its relatives. We established neomycin resistance as a second selectable marker by designing a plasmid that confers neomycin resistance in Ms. mazei.

  16. Simultaneous stable expression of neomycin phosphotransferase and green fluorescence protein genes in Trypanosoma cruzi.

    Science.gov (United States)

    dos Santos, W G; Buck, G A

    2000-12-01

    The ribosomal RNA (rRNA) gene promoter was used to construct plasmid vectors that simultaneously express multiple exogenous genes in Trypanosoma cruzi. Vector pBSPANEO expresses neomycin phosphotransferase, and pPAGFPAN expresses both green fluorescent protein and neomycin phosphotransferase from a single promoter. Both vectors require the presence of the rRNA promoter for stable transfection; epimastigotes transfected with pPAGFPAN strongly fluoresced due to green fluorescent protein expression. Intact plasmids were rescued from the T. cruzi-transfected population after >8 mo of culture, indicating stable replication of these vectors. Vectors were integrated into the rRNA locus by homologous recombination and into other loci, presumably by illegitimate recombination. Parasites bearing tandem concatamers of plasmids were also found among the transfectants. Transfectants expressing green fluorescent protein showed a bright green fluorescence distributed throughout the cell. Fluorescence was also detected in amastigotes after infection of mammalian cells with transfected parasites, indicating that the rRNA promoter can drive efficient expression of these reporter genes in multiple life-cycle stages of the parasite. Expression of the heterologous genes was detected after passage in mice or in the insect vector. These vectors will be useful for the genetic dissection of T. cruzi biology and pathogenesis.

  17. Novel plasmid conferring kanamycin and tetracycline resistance in the turkey-derived Campylobacter jejuni strain 11601MD.

    Science.gov (United States)

    Crespo, M D; Altermann, E; Olson, J; Miller, W G; Chandrashekhar, K; Kathariou, S

    2016-07-01

    In Campylobacter spp., resistance to the antimicrobials kanamycin and tetracycline is frequently associated with plasmid-borne genes. However, relatively few plasmids of Campylobacter jejuni have been fully characterized to date. A novel plasmid (p11601MD; 44,095nt) harboring tet(O) was identified in C. jejuni strain 11601MD, which was isolated from the jejunum of a turkey produced conventionally in North Carolina. Analysis of the p11601MD sequence revealed the presence of a high-GC content cassette with four genes that included tet(O) and a putative aminoglycoside transferase gene (aphA-3) highly similar to kanamycin resistance determinants. Several genes putatively involved in conjugative transfer were also identified on the plasmid. These findings will contribute to a better understanding of the distribution of potentially self-mobilizing plasmids harboring antibiotic resistance determinants in Campylobacter spp. from turkeys and other sources.

  18. Effect of antibiotic cefotaxime and kanamycin on callus formation and plantlet regeneration from leaves and callus of mangosteen

    Directory of Open Access Journals (Sweden)

    Rugpheug, R.

    2002-10-01

    Full Text Available In order to get rid of contamination from Agrobacterium tumefaciens, the bacterium employed in gene transformation, various kinds and concentrations of antibiotics were added singly or in combinations. In this investigation, concentrations of cefotaxime and kanamycin were examined for callus formation and regenerability from leaves and callus. The results showed that cefotaxime at the concentration of up to 300 mg/l gave a non-significant difference in callus formation. In the case of direct shoot bud formation, concentration over 100 mg/l drastically reduced percentage of leaf-forming shoot buds. The calli which were cultured continuously in 300 mg/l cefotaxime-containing medium for 6 passages gave callus forming shoot buds of 35%. Higher concentration of cefotaxime drastically decreased bud formation. In the case of kanamycin, callus could be induced and maintained in the medium supplemented with a lower concentration than cefotaxime. However, the callus could not be maintained after 3 subculturings.

  19. Preparation and Microbiological Evaluation of Amphiphilic Kanamycin-Lipoamino Acid Ion-Pairs

    Directory of Open Access Journals (Sweden)

    Rosario Pignatello

    2014-05-01

    Full Text Available Amphiphilic ion-pairs of kanamycin (KAN were prepared by evaporation of a water-ethanol co-solution of KAN base and a lipoamino acid bearing a 12-carbon atoms alkyl side chain (LAA12, at different molar ratios. Infrared spectroscopy confirmed the structure of ion-pairs, while differential scanning calorimetry (DSC and powder X-ray diffractometry (PXRD studies supported the formation of new saline species with a different crystalline structure than the starting components. The solubility pattern shown in a range of both aqueous and organic solvents confirmed that the ion-pairs possess an amphiphilic character. The LAA12 counter-ion showed not to improve the antibacterial activity of KAN, suggesting that such chemical strategy is not able to favor the penetration of this drug inside the bacteria cells. Nevertheless, a slight improving, i.e., a one-fold dilution, was observed in E. coli. The present study can also serve as the basis for a further evaluation of LAA ion-pairing of antibiotics, as a means to improve the loading of hydrophilic drugs into lipid-based nanocarriers.

  20. Subinhibitory concentration of kanamycin induces the Pseudomonas aeruginosa type VI secretion system.

    Directory of Open Access Journals (Sweden)

    Cerith Jones

    Full Text Available Pseudomonas aeruginosa is a Gram-negative bacterium found in natural environments including plants, soils and warm moist surfaces. This organism is also in the top ten of nosocomial pathogens, and prevalent in cystic fibrosis (CF lung infections. The ability of P. aeruginosa to colonize a wide variety of environments in a lasting manner is associated with the formation of a resistant biofilm and the capacity to efficiently outcompete other microorganisms. Here we demonstrate that sub-inhibitory concentration of kanamycin not only induces biofilm formation but also induces expression of the type VI secretion genes in the H1-T6SS cluster. The H1-T6SS is known for its role in toxin production and bacterial competition. We show that the antibiotic induction of the H1-T6SS only occurs when a functional Gac/Rsm pathway is present. These observations may contribute to understand how P. aeruginosa responds to antibiotic producing competitors. It also suggests that improper antibiotic therapy may enhance P. aeruginosa colonization, including in the airways of CF patients.

  1. The Effect of Kanamycin and Tetracycline on Growth and Photosynthetic Activity of Two Chlorophyte Algae

    Directory of Open Access Journals (Sweden)

    Khawaja Muhammad Imran Bashir

    2016-01-01

    Full Text Available Antibiotics are routinely used in microalgae culture screening, stock culture maintenance, and genetic transformation. By studying the effect of antibiotics on microalgae growth, we can estimate the least value to inhibit growth of undesired pathogens in algal culture. We studied the effect of kanamycin and tetracycline on the growth and photosynthetic activity of two chlorophyte microalgae, Dictyosphaerium pulchellum and Micractinium pusillum. We measured CFU mL−1 on agar plates, optical density, fluorescence yields, and photosynthetic inhibition. Our results showed a significant effect of kan and tet on the tested microalgae species except tet, which showed a minor effect on M. pusillum. Both antibiotics are believed to interact with the protein synthesis machinery; hence, the inhibitory effect of the tested antibiotics was further confirmed by isolation and quantification of the whole cell protein. A significant reduction in protein quantity was observed at concentrations more than 5 mg L−1, except M. pusillum, which showed only a slight reduction in protein quantity even at the maximum tested concentration of tet (30 mg L−1. This study can further aid in aquaculture industry, for the maintenance of the microalgae stock cultures and it can also help the microalgae genetic engineers in the construction of molecular markers.

  2. Computer-based design of novel HIV-1 entry inhibitors: neomycin conjugated to arginine peptides at two specific sites.

    Science.gov (United States)

    Berchanski, Alexander; Lapidot, Aviva

    2009-03-01

    Aminoglycoside-arginine conjugates (AAC and APAC) are multi-target inhibitors of human immunodeficiency virus type-1 (HIV-1). Here, we predict new conjugates of neomycin with two arginine peptide chains binding at specific sites on neomycin [poly-arginine-neomycin-poly-arginine (PA-Neo-PA)]. The rationale for the design of such compounds is to separate two short arginine peptides with neomycin, which may extend the binding region of the CXC chemokine receptor type 4 (CXCR4). We used homology models of CXCR4 and unliganded envelope glycoprotein 120 (HIV-1(IIIB) gp120) and docked PA-Neo-PAs and APACs to these using a multistep docking procedure. The results indicate that PA-Neo-PAs spread over two negatively charged patches of CXCR4. PA-Neo-PA-CXCR4 complexes are energetically more favorable than AACs/APAC-CXCR4 complexes. Notably, our CXCR4 model and docking procedure can be applied to predict new compounds that are either inhibitors of gp120-CXCR4 binding without affecting stromal cell-derived factor 1 alpha (SDF-1 alpha) chemotaxis activity, or inhibitors of SDF-1 alpha-CXCR4 binding resulting in an anti-metastasis effect. We also predict that PA-Neo-PAs and APACs can interfere with CD4-gp120 binding in unliganded conformation.

  3. Fecal excretion pattern of bile acids in rats fed high fat diets and neomycin in induced colon tumorigenesis.

    Science.gov (United States)

    Panda, S K; Broitman, S A

    1999-09-06

    Neomycin augments colon tumorigenesis in 1,2 - dimethylhydrazine treated rats fed polyunsaturated fat diet and decreases fecal cholic acid excretion, while it inhibits tumorigenesis with increased cholic acid and decreased deoxycholic acid excretions in rats fed high cholesterol diet. Participation of other fecal bile acids seems to be insignificant in relation to colon carcinogenesis.

  4. Neomycin B inhibits splicing of the td intron indirectly by interfering with translation and enhances missplicing in vivo.

    Science.gov (United States)

    Waldsich, C; Semrad, K; Schroeder, R

    1998-12-01

    The aminoglycoside antibiotic neomycin B inhibits translation in prokaryotes and interferes with RNA-protein interactions in HIV both in vivo and in vitro. Hitherto, inhibition of ribozyme catalysis has only been observed in vitro. We therefore monitored the activity of neomycin B and several other aminoglycoside antibiotics on splicing of the T4 phage thymidylate synthase (td) intron in vivo. All antibiotics tested inhibited splicing, even chloramphenicol, which does not inhibit splicing in vitro. Splicing of the td intron in vivo requires translation for proper folding of the pre-mRNA. In the absence of translation, two interactions between sequences in the upstream exon and the 5' and 3' splice sites trap the pre-mRNA in splicing-incompetent conformations. Their disruption by mutations rendered splicing less dependent on translation and also less sensitive to neomycin B. Intron splicing was affected by neither neomycin B nor gentamicin in Escherichia coli strains carrying antibiotic-resistance genes that modify the ribosomal RNA. Taken together, this demonstrates that in vivo splicing of td intron is not directly inhibited by aminoglycosides, but rather indirectly by their interference with translation. This was further confirmed by assaying splicing of the Tetrahymena group I intron, which is inserted in the E. coli 23 S rRNA and, thus, not translated. Furthermore, neomycin B, paromomycin, and streptomycin enhanced missplicing in antibiotic-sensitive strains. Missplicing is caused by an alternative structural element containing a cryptic 5' splice site, which serves as a substrate for the ribozyme. Our results demonstrate that aminoglycoside antibiotics display different effects on ribozymes in vivo and in vitro.

  5. Protective role of L-ascorbic acid, N-acetylcysteine and apocynin on neomycin-induced hair cell loss in zebrafish.

    Science.gov (United States)

    Wu, Chia-Yen; Lee, Han-Jung; Liu, Chi-Fang; Korivi, Mallikarjuna; Chen, Hwei-Hsien; Chan, Ming-Huan

    2015-03-01

    Hair cells are highly sensitive to environmental insults and other therapeutic drugs. The adverse effects of drugs such as aminoglycosides can cause hair cell death and lead to hearing loss and imbalance. The objective of the present study was to evaluate the protective activity of L-ascorbic acid, N-acetylcysteine (NAC) and apocynin on neomycin-induced hair cell damage in zebrafish (Danio rerio) larvae at 5 days post fertilization (dpf). Results showed that the loss of hair cells within the neuromasts of the lateral lines after neomycin exposure was evidenced by a significantly lower number of neuromasts labeled with fluorescent dye FM1-43FX observed under a microscope. Co-administration with L-ascorbic acid, NAC and apocynin protected neomycin-induced hair cell loss within the neuromasts. Moreover, these three compounds reduced the production of reactive oxygen species (ROS) in neuromasts exposed to neomycin, indicating that their antioxidant action is involved. In contrast, the neuromasts were labeled with specific fluorescent dye Texas-red conjugated with neomycin to detect neomycin uptake. Interestingly, the uptake of neomycin into hair cells was not influenced by these three antioxidant compounds. These data imply that prevention of hair cell damage against neomycin by L-ascorbic acid, NAC and apocynin might be associated with inhibition of excessive ROS production, but not related to modulating neomycin uptake. Our findings conclude that L-ascorbic acid, NAC and apocynin could be used as therapeutic drugs to protect aminoglycoside-induced listening impairment after further confirmatory studies.

  6. Guanidinylated neomycin mediates heparan sulfate-dependent transport of active enzymes to lysosomes.

    Science.gov (United States)

    Sarrazin, Stéphane; Wilson, Beth; Sly, William S; Tor, Yitzhak; Esko, Jeffrey D

    2010-07-01

    Guanidinylated neomycin (GNeo) can transport bioactive, high molecular weight cargo into the interior of cells in a process that depends on cell surface heparan sulfate proteoglycans. In this report, we show that GNeo-modified quantum dots bind to cell surface heparan sulfate, undergo endocytosis and eventually reach the lysosomal compartment. An N-hydroxysuccinimide activated ester of GNeo (GNeo-NHS) was prepared and conjugated to two lysosomal enzymes, beta-D-glucuronidase (GUS) and alpha-L-iduronidase. Conjugation did not interfere with enzyme activity and enabled binding of the enzymes to heparin-Sepharose and heparan sulfate on primary human fibroblasts. Cells lacking the corresponding lysosomal enzyme took up sufficient amounts of the conjugated enzymes to restore normal turnover of glycosaminoglycans. The high capacity of proteoglycan-mediated uptake suggests that this method of delivery might be used for enzyme replacement or introduction of foreign enzymes into cells.

  7. Spectroscopic probe of the competitive binding of ethidium bromide and neomycin to DNA

    Science.gov (United States)

    Pal, Medini Kanta; Ghosh, Jimut Kanti

    1995-03-01

    The three spectroscopic changes of ethidium bromide (EB) on its binding to DNA, namely red-shift of the νmax, enhancement of fluorescence and induced dichroism are utilized to study the competitive binding of neomycin (NMC) and EB to DNA. Reversion of νmax, decrease in fluorescence and reduction of dichroism of DNA-EB on addition of NMC shows that the binding of NMC and EB to DNA is competitive in nature, over a limited concentration of the polymer. The binding constant of EB-DNA falls from 4.00 × 10 6 to 2.27 × 10 4 1 mol -1 in the presence of added NMC.

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

    Institute of Scientific and Technical Information of China (English)

    Hong LIU; Da-lian DING; Hai-yan JIANG; Xue-wen WU; Richard SALVI; Hong SUN

    2011-01-01

    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-KM-induced 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.

  9. Diagnosis of Drug Resistance to Fluoroquinolones, Amikacin, Capreomycin, Kanamycin and Ethambutol with Genotype MTBDRsl Assay: a Meta-Analysis.

    Science.gov (United States)

    Mao, Xiaolu; Ke, Zunqiong; Shi, Xiaoyan; Liu, Shuiyi; Tang, Beibei; Wang, Jin; Huang, Hao

    2015-01-01

    The Genotype MTBDRsl is a new-generation PCR-based line-probe assay for rapid identification of the resistance to the second-line antituberculosis drugs with a single strip. The aim of this meta-analysis was to evaluate the performance of Genotype MTBDRsl in detecting drug resistance to fluoroquinolones, amikacin, capreomycin, kanamycin and ethambutol in comparison with the phenotypic drug susceptibility test. We searched Pubmed, Embase and the Cochrane Library and calculated the sensitivity, the specificity, the positive likelihood ratio (PLR), negative likelihood ratio (NLR), diagnostic odds ratio (DOR), corresponding 95% confidence interval (CI), and the area under the summary receiver operating characteristic (SROC) curves (AUC), and tested heterogeneity in accuracy estimates with the Spearman correlation coefficient and Chi-square. The summarized sensitivity (95% CI), specificity (95% CI), and AUC (standard error) were 0.869 (0.847-0.890), 0.973 (0.965-0.979) and 0.9690 (0.0188) for fluoroquinolones, 0.868 (0.829-0.900), 0.998 (0.994-0.999) and 0.9944 (0.0050) for amikacin, 0.879 (0.838-0.914), 0.970 (0.958-0.978) and 0.9791 (0.0120) for capreomycin, 0.501 (0.461-0.541), 0.991 (0.983-0.996) and 0.9814 (0.0114) for kanamycin and 0.686 (0.663-0.709), 0.871 (0.852-0.888) and 0.7349 (0.0639) for ethambutol, respectively. The genotype MTBDRsl demonstrate excellent accuracy for detecting drug resistance to fluoroquinolones, amikacin, and capreomycin, but it may not be an appropriate choice for detection of kanamycin and ethambutol. © 2015 by the Association of Clinical Scientists, Inc.

  10. Synthesis and properties of vitamin E analog-conjugated neomycin for delivery of RNAi drugs to liver cells.

    Science.gov (United States)

    Iwata, Rintaro; Nakayama, Futoshi; Hirochi, Sakie; Sato, Kazuki; Piao, Wenying; Nishina, Kazutaka; Yokota, Takanori; Wada, Takeshi

    2015-02-15

    RNA interference (RNAi) is a promising tool to regulate gene expression by external double stranded RNAs (dsRNAs) such as siRNAs. As an efficient method to deliver siRNAs to liver cells, we propose a novel strategy using vitamin E (VE)-conjugated neomycin derivatives. With the aim of delivering RNAi-based drugs to liver cells, several tripod-type and prodrug-type neomycin derivatives were synthesized, all of which were thermodynamically stabilized RNA duplexes. The prodrug-type derivative 7 and the tripod-type derivative 10 were delivered to liver cancer cells and successfully induced RNAi activity. These results indicated the potential use of natural aminoglycosides as carriers of RNAi drugs.

  11. Neomycin-loaded poly(styrene sulfonic acid-co-maleic acid) (PSSA-MA)/polyvinyl alcohol (PVA) ion exchange nanofibers for wound dressing materials.

    Science.gov (United States)

    Nitanan, Todsapon; Akkaramongkolporn, Prasert; Rojanarata, Theerasak; Ngawhirunpat, Tanasait; Opanasopit, Praneet

    2013-05-01

    In this study, poly(styrene sulfonic acid-co-maleic acid) (PSSA-MA) blended with polyvinyl alcohol (PVA) was electrospun and then subjected to thermal crosslinking to produce PSSA-MA/PVA ion exchange nanofiber mats. The cationic drug neomycin (0.001, 0.01, and 0.1%, w/v) was loaded onto the cationic exchange fibers. The amount of neomycin loaded and released and the cytotoxicity of the fiber mats were analyzed. In vivo wound healing tests were also performed in Wistar rats. The results indicated that the diameters of the fibers were on the nanoscale (250 ± 21 nm). The ion exchange capacity (IEC) value and the percentage of water uptake were 2.19 ± 0.1 mequiv./g-dry fibers and 268 ± 15%, respectively. The loading capacity was increased upon increasing the neomycin concentration. An initial concentration of 0.1% (w/v) neomycin (F3) showed the highest loading capacity (65.7 mg/g-dry fibers). The neomycin-loaded nanofiber mats demonstrated satisfactory antibacterial activity against both Gram-positive and Gram-negative bacteria, and an in vivo wound healing test revealed that these mats performed better than gauze and blank nanofiber mats in decreasing acute wound size during the first week after tissue damage. In conclusion, the antibacterial neomycin-loaded PSSA-MA/PVA cationic exchange nanofiber mats have the potential for use as wound dressing materials.

  12. Efficacy of neomycin sulfate water medication on the control of mortality associated with colibacillosis in growing turkeys.

    Science.gov (United States)

    Marrett, L E; Robb, E J; Frank, R K

    2000-01-01

    The objective of this investigation was to evaluate the efficacy, safety, and toxicity of neomycin sulfate (Neomix 325) water medication to control mortality associated with colibacillosis (Escherichia coli) in growing turkeys. One efficacy trial was conducted at five locations; each location included 2,880 sexed 21-d-old turkey poults that were naturally challenged with litter from turkey flocks that had colibacillosis. Between 5 and 7 d after challenge, and when mortality had reached 0.5%, poults were randomized within sex into three treatment groups of 0, 11, or 22 mg neomycin sulfate/kg body weight. In each location, each treatment was replicated 12 times with 40 poults per sex per replicate. All treatments were administered in the drinking water for 5 d. The pivotal decision criterion was mortality. Mortality was defined as 1) supported mortality (SM): positive microbial culture for E. coli and gross lesions, 2) diagnosed mortality (DM): diagnosed as associated with E. coli but not supported by lesions or positive microbiological cultures, 3) overall mortality (OM): mortality associated with E. coli or other microorganisms and miscellaneous reasons such as accidents (trampling or suffocations). Performance data (growth and feed utilization) also were measured and are reported without statistical analysis. Results from this efficacy study clearly demonstrated the effectiveness of neomycin sulfate against E. coli as measured by a reduction in mortality. In the target animal safety and toxicity study (done in conjunction with the efficacy study), neomycin sulfate in the drinking water at 66, 110, or 220 mg/kg per d for 15 d had no observable adverse effects on poult performance, as measured by feed or water consumption, body weight, gross pathology, or mortality.

  13. Improved methods in Agrobacterium-mediated transformation of almond using positive (mannose/pmi) or negative (kanamycin resistance) selection-based protocols.

    Science.gov (United States)

    Ramesh, Sunita A; Kaiser, Brent N; Franks, Tricia; Collins, Graham; Sedgley, Margaret

    2006-08-01

    A protocol for Agrobacterium-mediated transformation with either kanamycin or mannose selection was developed for leaf explants of the cultivar Prunus dulcis cv. Ne Plus Ultra. Regenerating shoots were selected on medium containing 15 muM kanamycin (negative selection), while in the positive selection strategy, shoots were selected on 2.5 g/l mannose supplemented with 15 g/l sucrose. Transformation efficiencies based on PCR analysis of individual putative transformed shoots from independent lines relative to the initial numbers of leaf explants tested were 5.6% for kanamycin/nptII and 6.8% for mannose/pmi selection, respectively. Southern blot analysis on six randomly chosen PCR-positive shoots confirmed the presence of the nptII transgene in each, and five randomly chosen lines identified to contain the pmi transgene by PCR showed positive hybridisation to a pmi DNA probe. The positive (mannose/pmi) and the negative (kanamycin) selection protocols used in this study have greatly improved transformation efficiency in almond, which were confirmed with PCR and Southern blot. This study also demonstrates that in almond the mannose/pmi selection protocol is appropriate and can result in higher transformation efficiencies over that of kanamycin/nptII selection protocols.

  14. Neomycin and pentagalloyl glucose enhanced cross-linking for elastin and glycosaminoglycans preservation in bioprosthetic heart valves.

    Science.gov (United States)

    Tripi, Daniel R; Vyavahare, Naren R

    2014-01-01

    Glutaraldehyde cross-linked bioprosthetic heart valves fail within 12-15 years of implantation due to limited durability. Glutaraldehyde does not adequately stabilize extracellular matrix components such as glycosaminoglycans and elastin, and loss of these components could be a major cause of degeneration of valve after implantation. We have shown earlier that neomycin-based cross-linking stabilizes glycosaminoglycans in the tissue but fails to stabilize elastin component. Here, we report a new treatment where neomycin and pentagalloyl glucose (PGG) were incorporated into glutaraldehyde cross-linking neomycin-PGG-Glutaraldehyde (NPG) to stabilize both glycosaminoglycans and elastin in porcine aortic valves. In vitro studies demonstrated a marked increase in extracellular matrix stability against enzymatic degradation after cross-linking and 10 month storage in NPG group when compared to glutaraldehyde controls. Tensile properties showed increased lower elastic modulus in both radial and circumferential directions in NPG group as compared to glutaraldehyde, probably due to increased elastin stabilization with no changes in upper elastic modulus and extensibility. The enhanced extracellular matrix stability was further maintained in NPG-treated tissues after rat subdermal implantation for three weeks. NPG group also showed reduced calcification when compared to glutaraldehyde controls. We conclude that NPG cross-linking would be an excellent alternative to glutaraldehyde cross-linking of bioprosthetic heart valves to improve its durability.

  15. [Development and application of indirect competitive enzyme immunoassay for detection of neomycin in milk].

    Science.gov (United States)

    Burkin, M A; Gal'vidis, I A

    2011-01-01

    As a result of immunization of rabbits with neomycin B (N M) conjugated to periodate-oxidized transferrin, polyclonal antibodies were generated and used to develop an indirect competitive enzyme-linked immunosorbent assay (ELISA) of NM. Several heterologous conjugates, namely, glutaraldehyde (GA)-polymerized NM, gelatin-ribostamycin (sp), and gelatin-NM (ga) were used as coating antigens in different ELISA variants for quantification of NM in milk. These variants were characterized by different dynamic ranges and detection limits of 1.0, 0.1, and 0.01 ng/ml, respectively. Maximum residue level (MRL) of this antibiotic in milk accepted in the EU can be detected without any special pretreatment at a 100-fold sample dilution in the least sensitive assay variant. The mean recovery rate from NM-spiked milk containing 1.5-10% fat was 111.7% and ranged from 84 to 125.2%. We found that 57 of 106 tested milk samples contained NM at concentrations higher than 100 ng/ml. In ten percent of cases (11/1 06), the residual level of this antibiotic was greater than 500 ng/ml. In one case, the M RL was exceeded (1690 ng/ml). The assay developed in this study is specific shows no cross-reactivity with other veterinary aminoglycosides, has a good sensitivity reserve, and can serve as an effective tool to monitor the NM content in milk stuff.

  16. Amplification strategy based on gold nanoparticle-decorated carbon nanotubes for neomycin immunosensors.

    Science.gov (United States)

    Zhu, Ye; Son, Jung Ik; Shim, Yoon-Bo

    2010-11-15

    A novel amperometric immunosensor with an enhanced sensitivity for the detection of neomycin (Neo) was prepared by covalently immobilizing a monoclonal Neo antibody onto a new conducting polymer, poly-[2,5-di-(2-thienyl)-1H-pyrrole-1-(p-benzoic acid)] (pDPB), as a sensor probe. The probe was used to detect Neo in a sandwich-type approach, where the secondary antibody was attached to gold nanoparticle-decorated multi-wall carbon nanotubes labeled with hydrazine (Hyd-MWCNT(AuNP)-Ab(2)). Hydrazine on the conjugate served as a catalyst for the reduction of hydrogen peroxide, and the catalytic current was monitored at -0.45 V vs. Ag/AgCl. The performance of the immunosensor with and without AuNPs on the probe and the conjugate was compared. The parameters affecting the immunosensor response in terms of antibody dilution ratio, incubation time, pH, applied potential, and temperature were optimized. A linear dynamic range for Neo analysis was obtained between 10 ng/mL and 250 ng/mL with a detection limit of 6.76 ± 0.17 ng/mL. The proposed immunosensor was successfully applied to detect Neo content in real meat samples.

  17. Influence of linker length and composition on enzymatic activity and ribosomal binding of neomycin dimers.

    Science.gov (United States)

    Watkins, Derrick; Kumar, Sunil; Green, Keith D; Arya, Dev P; Garneau-Tsodikova, Sylvie

    2015-07-01

    The human and bacterial A site rRNA binding as well as the aminoglycoside-modifying enzyme (AME) activity against a series of neomycin B (NEO) dimers is presented. The data indicate that by simple modifications of linker length and composition, substantial differences in rRNA selectivity and AME activity can be obtained. We tested five different AMEs with dimeric NEO dimers that were tethered via triazole, urea, and thiourea linkages. We show that triazole-linked dimers were the worst substrates for most AMEs, with those containing the longer linkers showing the largest decrease in activity. Thiourea-linked dimers that showed a decrease in activity by AMEs also showed increased bacterial A site binding, with one compound (compound 14) even showing substantially reduced human A site binding. The urea-linked dimers showed a substantial decrease in activity by AMEs when a conformationally restrictive phenyl linker was introduced. The information learned herein advances our understanding of the importance of the linker length and composition for the generation of dimeric aminoglycoside antibiotics capable of avoiding the action of AMEs and selective binding to the bacterial rRNA over binding to the human rRNA.

  18. Dual Targeting of Intracellular Pathogenic Bacteria with a Cleavable Conjugate of Kanamycin and an Antibacterial Cell-Penetrating Peptide.

    Science.gov (United States)

    Brezden, Anna; Mohamed, Mohamed F; Nepal, Manish; Harwood, John S; Kuriakose, Jerrin; Seleem, Mohamed N; Chmielewski, Jean

    2016-08-31

    Bacterial infection caused by intracellular pathogens, such as Mycobacterium, Salmonella, and Brucella, is a burgeoning global health epidemic that necessitates urgent action. However, the therapeutic value of a number of antibiotics, including aminoglycosides, against intracellular pathogenic bacteria is compromised due to their inability to traverse eukaryotic membranes. For this significant problem to be addressed, a cleavable conjugate of the antibiotic kanamycin and a nonmembrane lytic, broad-spectrum antimicrobial peptide with efficient mammalian cell penetration, P14LRR, was prepared. This approach allows kanamycin to enter mammalian cells as a conjugate linked via a tether that breaks down in the reducing environment within cells. Potent antimicrobial activity of the P14KanS conjugate was demonstrated in vitro, and this reducible conjugate effectively cleared intracellular pathogenic bacteria within macrophages more potently than that of a conjugate lacking the disulfide moiety. Notably, successful clearance of Mycobacterium tuberculosis within macrophages was observed with the dual antibiotic conjugate, and Salmonella levels were significantly reduced in an in vivo Caenorhabditis elegans model.

  19. Kaposi's sarcoma-associated herpesvirus-positive primary effusion lymphoma tumor formation in NOD/SCID mice is inhibited by neomycin and neamine blocking angiogenin's nuclear translocation.

    Science.gov (United States)

    Bottero, Virginie; Sadagopan, Sathish; Johnson, Karen E; Dutta, Sujoy; Veettil, Mohanan Valiya; Chandran, Bala

    2013-11-01

    Angiogenin (ANG) is a 14-kDa multifunctional proangiogenic secreted protein whose expression level correlates with the aggressiveness of several tumors. We observed increased ANG expression and secretion in endothelial cells during de novo infection with Kaposi's sarcoma-associated herpesvirus (KSHV), in cells expressing only latency-associated nuclear antigen 1 (LANA-1) protein, and in KSHV latently infected primary effusion lymphoma (PEL) BCBL-1 and BC-3 cells. Inhibition of phospholipase Cγ (PLCγ) mediated ANG's nuclear translocation by neomycin, an aminoglycoside antibiotic (not G418-neomicin), resulted in reduced KSHV latent gene expression, increased lytic gene expression, and increased cell death of KSHV(+) PEL and endothelial cells. ANG detection in significant levels in KS and PEL lesions highlights its importance in KSHV pathogenesis. To assess the in vivo antitumor activity of neomycin and neamine (a nontoxic derivative of neomycin), BCBL-1 cells were injected intraperitoneally into NOD/SCID mice. We observed significant extended survival of mice treated with neomycin or neamine. Markers of lymphoma establishment, such as increases in animal body weight, spleen size, tumor cell spleen infiltration, and ascites volume, were observed in nontreated animals and were significantly diminished by neomycin or neamine treatments. A significant decrease in LANA-1 expression, an increase in lytic gene expression, and an increase in cleaved caspase-3 were also observed in neomycin- or neamine-treated animal ascitic cells. These studies demonstrated that ANG played an essential role in KSHV latency maintenance and BCBL-1 cell survival in vivo, and targeting ANG function by neomycin/neamine to induce the apoptosis of cells latently infected with KSHV is an attractive therapeutic strategy against KSHV-associated malignancies.

  20. Normal phenotype in conditional androgen receptor (AR) exon 3-floxed neomycin-negative male mice.

    Science.gov (United States)

    Rana, Kesha; Clarke, Michele V; Zajac, Jeffrey D; Davey, Rachel A; MacLean, Helen E

    2014-01-01

    Androgens (testosterone and dihydrotestosterone) acting via the androgen receptor (AR) are required for male sexual differentiation, and also regulate the development of many other tissues including muscle, fat and bone. We previously generated an AR(lox) mouse line with exon 3 of the AR gene targeted by loxP sites. The deletion of exon 3 is in-frame, so only the DNA binding-dependent actions of the AR are deleted, but non-DNA binding-dependent actions are retained. This line also contained an antibiotic resistance selection cassette, neomycin (neo) in intron 3, which was also flanked by loxP sites. Hemizygous AR(lox) male mice demonstrated a phenotype of hyperandrogenization, with increased mass of androgen-dependent tissues. We hypothesized that this hyperandrogenization was likely to be due to the presence of the neo cassette. In this study, we have generated an AR(lox) neo-negative mouse line, using the EIIa-cre deleter mouse line to remove the neo cassette. Hemizygous AR(lox) neo-negative male mice have a normal phenotype, with normal body mass and normal mass of androgen-dependent tissues including the testis, seminal vesicles, kidney, spleen, heart and retroperitoneal fat. This neo-negative exon 3-targeted mouse line is the only floxed AR mouse line available to study the DNA binding-dependent actions of the AR in a tissue-specific manner, and is suitable for investigation in all tissues. This study demonstrates the importance of removing the selection cassette, which can potentially alter the phenotype of floxed mouse lines even in the absence of detectable effects on target gene expression.

  1. RNA-seq analysis of the effect of kanamycin and the ABC transporter AtWBC19 on Arabidopsis thaliana seedlings reveals changes in metal content.

    Directory of Open Access Journals (Sweden)

    Ayalew Mentewab

    Full Text Available Plants are exposed to antibiotics produced by soil microorganisms, but little is known about their responses at the transcriptional level. Likewise, few endogenous mechanisms of antibiotic resistance have been reported. The Arabidopsis thaliana ATP Binding Cassette (ABC transporter AtWBC19 (ABCG19 is known to confer kanamycin resistance, but the exact mechanism of resistance is not well understood. Here we examined the transcriptomes of control seedlings and wbc19 mutant seedlings using RNA-seq analysis. Exposure to kanamycin indicated changes in the organization of the photosynthetic apparatus, metabolic fluxes and metal uptake. Elemental analysis showed a 60% and 80% reduction of iron uptake in control and wbc19 mutant seedlings respectively, upon exposure to kanamycin. The drop in iron content was accompanied by the upregulation of the gene encoding for FERRIC REDUCTION OXIDASE 6 (FRO6 in mutant seedlings but not by the differential expression of other transport genes known to be induced by iron deficiency. In addition, wbc19 mutants displayed a distinct expression profile in the absence of kanamycin. Most notably the expression of several zinc ion binding proteins, including ZINC TRANSPORTER 1 PRECURSOR (ZIP1 was increased, suggesting abnormal zinc uptake. Elemental analysis confirmed a 50% decrease of zinc content in wbc19 mutants. Thus, the antibiotic resistance gene WBC19 appears to also have a role in zinc uptake.

  2. Comparing amikacin and kanamycin-induced hearing loss in multidrug-resistant tuberculosis treatment under programmatic conditions in a Namibian retrospective cohort

    NARCIS (Netherlands)

    Sagwa, Evans L; Ruswa, Nunurai; Mavhunga, Farai; Rennie, Timothy; Leufkens, Hubert G M|info:eu-repo/dai/nl/075255049; Mantel-Teeuwisse, Aukje K|info:eu-repo/dai/nl/266775098

    2015-01-01

    BACKGROUND: Amikacin and kanamycin are mainly used for treating multidrug-resistant tuberculosis (MDR-TB), especially in developing countries where the burden of MDR-TB is highest. Their protracted use in MDR-TB treatment is known to cause dose-dependent irreversible hearing loss, requiring hearing

  3. Comparing amikacin and kanamycin-induced hearing loss in multidrug-resistant tuberculosis treatment under programmatic conditions in a Namibian retrospective cohort

    NARCIS (Netherlands)

    Sagwa, Evans L; Ruswa, Nunurai; Mavhunga, Farai; Rennie, Timothy; Leufkens, Hubert G M; Mantel-Teeuwisse, Aukje K

    2015-01-01

    BACKGROUND: Amikacin and kanamycin are mainly used for treating multidrug-resistant tuberculosis (MDR-TB), especially in developing countries where the burden of MDR-TB is highest. Their protracted use in MDR-TB treatment is known to cause dose-dependent irreversible hearing loss, requiring hearing

  4. Sulfonamide-Based Inhibitors of Aminoglycoside Acetyltransferase Eis Abolish Resistance to Kanamycin in Mycobacterium tuberculosis

    Energy Technology Data Exchange (ETDEWEB)

    Garzan, Atefeh; Willby, Melisa J.; Green, Keith D.; Gajadeera, Chathurada S.; Hou, Caixia; Tsodikov, Oleg V.; Posey, James E.; Garneau-Tsodikova, Sylvie

    2016-12-08

    A two-drug combination therapy where one drug targets an offending cell and the other targets a resistance mechanism to the first drug is a time-tested, yet underexploited approach to combat or prevent drug resistance. By high-throughput screening, we identified a sulfonamide scaffold that served as a pharmacophore to generate inhibitors of Mycobacterium tuberculosis acetyltransferase Eis, whose upregulation causes resistance to the aminoglycoside (AG) antibiotic kanamycin A (KAN) in Mycobacterium tuberculosis. Rational systematic derivatization of this scaffold to maximize Eis inhibition and abolish the Eis-mediated KAN resistance of M. tuberculosis yielded several highly potent agents. A crystal structure of Eis in complex with one of the most potent inhibitors revealed that the inhibitor bound Eis in the AG-binding pocket held by a conformationally malleable region of Eis (residues 28–37) bearing key hydrophobic residues. These Eis inhibitors are promising leads for preclinical development of innovative AG combination therapies against resistant TB.

  5. Decreased expression of humanized Fat-1 in porcine fetal fibroblasts following deletion of PGK-neomycin resistance.

    Science.gov (United States)

    Han, X J; Liang, H; Yun, T; Zhao, Y H; Zhang, M L; Zhao, L H; Li, R F; Li, X L

    2015-09-28

    The neomycin-resistance (neo(r)) gene is widely used as a selectable marker in eukaryotic expression vectors; however, its expression often affects that of target genes. Cre recombinase recognizes LoxP sites, leading to site-specific recombination and deletion of DNA and RNA between two LoxP sites. In the present study, a humanized Fat-1 gene (hFat-1) was generated by DNA Works and used to construct a pC-PGK-neo(r)-hfat-1 expression vector, in which PGK-neo(r) was flanked by two LoxP sites. The pC-PGK-neo(r)-hfat-1 plasmids were transfected into porcine fetal fibroblasts using liposomes, and three transgenic cell lines were obtained by culturing with 400 μg/mL G418 for 7 days. Next, these cell lines were transfected with a Cre recombinase expression plasmid, which contains a puromycin resistance gene, in order to delete neo(r), which was integrated into the genome. hFat-1-neo(r) negative cells were obtained following puromycin selection. Real-time quantitative polymerase chain reaction data indicated that neomycin-resistant cells had higher hFat-1 expression than neomycin-sensitive cells. High performance gas chromatography data suggested that the n-6/n-3 ratio was significantly lower in transfected cells than in wild-type cells. The n-6/n-3 ratio in Cre-treated hFat-1-transfected cells was higher than that in untreated cells, suggesting that deletion of PGK-neo(r) decreased hFat-1 expression.

  6. Neomycin-phenolic conjugates: polycationic amphiphiles with broad-spectrum antibacterial activity, low hemolytic activity and weak serum protein binding.

    Science.gov (United States)

    Findlay, Brandon; Zhanel, George G; Schweizer, Frank

    2012-02-15

    Here we present a proof-of-concept study, combining two known antimicrobial agents into a hybrid structure in order to develop an emergent cationic detergent-like interaction with the bacterial membrane. Six amphiphilic conjugates were prepared by copper (I)-catalyzed 1,3-dipolar cycloaddition between a neomycin B-derived azide and three alkyne-modified phenolic disinfectants. Three conjugates displayed good activity against a variety of clinically relevant Gram positive and Gram negative bacteria, including MRSA, without the high level of hemolysis or strong binding to serum proteins commonly observed with other cationic antimicrobial peptides and detergents.

  7. Structural elucidation of biologically active neomycin N-octyl derivatives in a regioisomeric mixture by means of liquid chromatography/ion trap time-of-flight mass spectrometry.

    Science.gov (United States)

    Giera, Martin; de Vlieger, Jon S B; Lingeman, Henk; Irth, Hubertus; Niessen, Wilfried M A

    2010-05-30

    Structural elucidation of six regioisomers of mono-N-octyl derivatized neomycin is achieved using MS(n) (up to n = 4) on an ion trap time-of-flight (IT-TOF) instrument equipped with electrospray ionization. The mixture of six derivatized neomycin analogues was generated by reductive amination in a shotgun synthetic approach. In parallel to the liquid chromatography/mass spectrometry (LC/MS) detection, the antibacterial activity of the neomycin regioisomers was tested by post-column addition of buffer and bacterial inocula, subsequent microfractionation of the resulting mixture, incubation, and finally a chemiluminescence-based bioactivity measurement based on the production of bacterial ATP. The MS-based high-resolution screening approach described can be applied in medicinal chemistry to help in designing and producing new antibiotic substances, which is particularly challenging due to the high functionality of most antibiotic substances, therefore requiring advanced (hyphenated) separation and detection techniques for compound mixtures.

  8. What a Difference an OH Makes: Conformational Dynamics as the Basis for the Ligand Specificity of the Neomycin-Sensing Riboswitch.

    Science.gov (United States)

    Duchardt-Ferner, Elke; Gottstein-Schmidtke, Sina R; Weigand, Julia E; Ohlenschläger, Oliver; Wurm, Jan-Philip; Hammann, Christian; Suess, Beatrix; Wöhnert, Jens

    2016-01-22

    To ensure appropriate metabolic regulation, riboswitches must discriminate efficiently between their target ligands and chemically similar molecules that are also present in the cell. A remarkable example of efficient ligand discrimination is a synthetic neomycin-sensing riboswitch. Paromomycin, which differs from neomycin only by the substitution of a single amino group with a hydroxy group, also binds but does not flip the riboswitch. Interestingly, the solution structures of the two riboswitch-ligand complexes are virtually identical. In this work, we demonstrate that the local loss of key intermolecular interactions at the substitution site is translated through a defined network of intramolecular interactions into global changes in RNA conformational dynamics. The remarkable specificity of this riboswitch is thus based on structural dynamics rather than static structural differences. In this respect, the neomycin riboswitch is a model for many of its natural counterparts.

  9. Comparative study of the effects of gentamicin, neomycin, streptomycin and ofloxacin antibiotics on sperm parameters and testis apoptosis in rats.

    Science.gov (United States)

    Khaki, Arash; Novin, Marefat Ghaffari; Khaki, Amir Afshin; Nouri, Mohammad; Sanati, Ehsan; Nikmanesh, Mahdad

    2008-07-01

    The aim of this study was to investigate the comparative effects of aminoglycosides and fluoroquinolones on testis apoptosis and sperm parameters in rats. Fifty male Wistar rats were randomly divided into control (n = 10) and experimental (n = 40) groups. The experimental groups subdivided into four groups often. Each received 5 mg kg(-1) (IP) gentamicin, 50 mg kg(-1) (IP) neomycin, 40 mg kg(-1) (IP) streptomycin and 72 mg kg(-1) (IP) ofloxacin daily for 14 days, respectively; however, the control group just received vehicle (IP). In the fourteenth day, rats were killed and sperm analyzed for sperm parameters. Testis tissues were also prepared for TUNEL assay for detection of apoptosis. There was a significant decrease in sperm count, viability and motility in all of experimental groups when compared with control group. Although in streptomycin group these parameters were less decreased than in the other experimental groups. The apoptotic cells were significantly increased in all experimental groups when compared with those seen in the controlled group. Gentamicin, neomycin and streptomycin and ofloxacin have negative effects on sperm parameters and testis apoptosis in rats. However, these side effects are less seen in the streptomycin group. Therefore, it is recommended that usage of this drug have fewer side effects on male fertility.

  10. Development, optimization and validation of a rapid colorimetric microplate bioassay for neomycin sulfate in pharmaceutical drug products.

    Science.gov (United States)

    Francisco, Fabiane Lacerda; Saviano, Alessandro Morais; Pinto, Terezinha de Jesus Andreoli; Lourenço, Felipe Rebello

    2014-08-01

    Microbiological assays have been used to evaluate antimicrobial activity since the discovery of the first antibiotics. Despite their limitations, microbiological assays are widely employed to determine antibiotic potency of pharmaceutical dosage forms, since they provide a measure of biological activity. The aim of this work is to develop, optimize and validate a rapid colorimetric microplate bioassay for the potency of neomycin in pharmaceutical drug products. Factorial and response surface methodologies were used in the development and optimization of the choice of microorganism, culture medium composition, amount of inoculum, triphenyltetrazolium chloride (TTC) concentration and neomycin concentration. The optimized bioassay method was validated by the assessment of linearity (range 3.0 to 5.0μg/mL, r=0.998 and 0.994 for standard and sample curves, respectively), precision (relative standard deviation (RSD) of 2.8% and 4.0 for repeatability and intermediate precision, respectively), accuracy (mean recovery=100.2%) and robustness. Statistical analysis showed equivalency between agar diffusion microbiological assay and rapid colorimetric microplate bioassay. In addition, microplate bioassay had advantages concerning the sensitivity of response, time of incubation, and amount of culture medium and solutions required.

  11. Validation of an enzyme-linked immunosorbent assay for qualitative screening of neomycin in muscle, liver, kidney, eggs and milk.

    Science.gov (United States)

    Solomun, B; Bilandzic, N; Varenina, I; Scortichini, G

    2011-01-01

    A rapid and sensitive enzyme-linked immunosorbent assay (ELISA) was used for the qualitative screening analysis of neomycin in food of animal origin (muscle, liver, kidney, eggs and milk) at levels corresponding to the European Union maximum residue limit (MRL) set for this substance. The method validation was performed according to the criteria of Commission Decision 2002/657/EC established for qualitative screening methods. In this regard, the following parameters were determined: detection capability (CCβ), specificity, detection limit (LOD), quantification limit (LOQ), recovery, precision, linearity and ruggedness. LODs ranged from 5.7 microg kg(-1) in kidney to 29.3 microg kg(-1) in milk; LOQs ranged from 11.4 microg kg(-1) in kidney to 59.7 microkg(-1) in eggs. The recoveries from spiked samples at the MRL, half the MRL and double the MRL levels ranged from 65.8% to 122.8%, with a coefficient of variation (CV) between 5.9% and 28.6%. The CCβ value was less than the MRL for all examined matrices. Moderate variations of some critical factors in the sample pretreatment for muscle, milk and eggs were deliberately introduced for ruggedness evaluation and had a slight but not statistically significant effect on method performance. The proposed method is suitable for qualitative screening analysis of neomycin in the above-mentioned food in conformity with current European Union performance requirements.

  12. Light-dependent repetitive Ca2+ spikes induced by extracellular application of neomycin in honeybee drone photoreceptors.

    Science.gov (United States)

    Walz, B; Zimmermann, B; Ukhanov, K

    2000-05-01

    Photoreceptor cells of the honeybee drone fire, in the presence of the polycationic aminoglycoside neomycin, repetitive slow spike-like potentials superimposed on the receptor potential plateau phase. We have used conventional intracellular recordings and microfluorometric intracellular Ca2+ measurements to characterize these spike potentials. We have shown that the spike frequency increases in a light-intensity-dependent manner. The spikes are fired only when light stimuli depolarize the cell from a resting potential of -50 to -60 mV to at least -40 to -45 mV; they are tetrodotoxin insensitive and blocked by the Ca2+ channel blockers Ni2+, Cd2+, omega-agatoxin TK, verapamil and methoxyverapamil. Depolarization of the photoreceptors with high extracellular K+ in the presence of neomycin in darkness does not generate spikes. Small intracellular Ca2+ oscillations superimposed on the plateau phase of the light-induced increase in intracellular free Ca2+ concentration have a similar temporal pattern as the spike-like potentials. We conclude that the spike-like potentials require stimulation by light and are generated by voltage-dependent Ca2+ channels localized on the soma of the photoreceptors, distal to the basal lamina.

  13. Autonomous assembly of synthetic oligonucleotides built from an expanded DNA alphabet. Total synthesis of a gene encoding kanamycin resistance

    Directory of Open Access Journals (Sweden)

    Kristen K. Merritt

    2014-10-01

    Full Text Available Background: Many synthetic biologists seek to increase the degree of autonomy in the assembly of long DNA (L-DNA constructs from short synthetic DNA fragments, which are today quite inexpensive because of automated solid-phase synthesis. However, the low information density of DNA built from just four nucleotide “letters”, the presence of strong (G:C and weak (A:T nucleobase pairs, the non-canonical folded structures that compete with Watson–Crick pairing, and other features intrinsic to natural DNA, generally prevent the autonomous assembly of short single-stranded oligonucleotides greater than a dozen or so.Results: We describe a new strategy to autonomously assemble L-DNA constructs from fragments of synthetic single-stranded DNA. This strategy uses an artificially expanded genetic information system (AEGIS that adds nucleotides to the four (G, A, C, and T found in standard DNA by shuffling hydrogen-bonding units on the nucleobases, all while retaining the overall Watson–Crick base-pairing geometry. The added information density allows larger numbers of synthetic fragments to self-assemble without off-target hybridization, hairpin formation, and non-canonical folding interactions. The AEGIS pairs are then converted into standard pairs to produce a fully natural L-DNA product. Here, we report the autonomous assembly of a gene encoding kanamycin resistance using this strategy. Synthetic fragments were built from a six-letter alphabet having two AEGIS components, 5-methyl-2’-deoxyisocytidine and 2’-deoxyisoguanosine (respectively S and B, at their overlapping ends. Gaps in the overlapped assembly were then filled in using DNA polymerases, and the nicks were sealed by ligase. The S:B pairs in the ligated construct were then converted to T:A pairs during PCR amplification. When cloned into a plasmid, the product was shown to make Escherichia coli resistant to kanamycin. A parallel study that attempted to assemble similarly sized genes

  14. In vitro protection of auditory hair cells by salicylate from the gentamicin-induced but not neomycin-induced cell loss.

    Science.gov (United States)

    Mazurek, Birgit; Lou, Xiangxin; Olze, Heidi; Haupt, Heidemarie; Szczepek, Agnieszka J

    2012-01-01

    Salicylate has been shown to protect animals and people from the gentamicin-induced hearing loss. The objective of our study was to determine if salicylate is otoprotective in vitro. In this fashion, we wanted to validate the use of explant culture system for future studies on the ototoxicity prevention. In addition, we wanted to find out if salicylate protects from the ototoxicity of other aminoglycosides. As a model, we used the membranous cochlear tissues containing the organ of Corti, spiral limbus and spiral ganglion neurons dissected from the cochleas of p3-p5 Wistar pups. The explants were divided into apical, medial and basal parts and cultured in presence or absence of 100μM gentamicin, 100μM neomycin and 5mM salicylate. Following the tissue fixation and staining with phalloidin-TRITC, the number of inner and outer hair cells (IHCs, OHCs) was scored under the fluorescent microscope. Presence of 5mM salicylate in explants cultures exposed to 100μM gentamicin significantly reduced the loss of IHCs and OHCs, as compared to explants exposed to gentamicin alone. In contrast, neomycin-induced auditory hair cell loss remained unaffected by the presence of salicylate. Our results corroborate earlier in vivo findings and validate the use of cochlear explants for future studies on ototoxicity and its prevention. Moreover, the inability of salicylate to prevent neomycin-induced ototoxicity implies possible differences between the mechanisms of auditory hair cell loss induced by gentamicin and neomycin.

  15. Electrochemical sensor using neomycin-imprinted film as recognition element based on chitosan-silver nanoparticles/graphene-multiwalled carbon nanotubes composites modified electrode.

    Science.gov (United States)

    Lian, Wenjing; Liu, Su; Yu, Jinghua; Li, Jie; Cui, Min; Xu, Wei; Huang, Jiadong

    2013-06-15

    A novel imprinted electrochemical sensor for neomycin recognition was developed based on chitosan-silver nanoparticles (CS-SNP)/graphene-multiwalled carbon nanotubes (GR-MWCNTs) composites decorated gold electrode. Molecularly imprinted polymers (MIPs) were synthesized by electropolymerization using neomycin as the template, and pyrrole as the monomer. The mechanism of the fabrication process and a number of factors affecting the activity of the imprinted sensor have been discussed and optimized. The characterization of imprinted sensor has been carried out by scanning electron microscope (SEM) and Fourier transform infrared spectroscopy (FTIR). The performance of the proposed imprinted sensor has been investigated using cyclic voltammetry (CV) and amperometry. Under the optimized conditions, the linear range of the sensor was from 9×10(-9)mol/L to 7×10(-6)mol/L, with the limit of detection (LOD) of 7.63×10(-9)mol/L (S/N=3). The film exhibited high binding affinity and selectivity towards the template neomycin, as well as good reproducibility and stability. Furthermore, the proposed sensor was applied to determine the neomycin in milk and honey samples based on its good reproducibility and stability, and the acceptable recovery implied its feasibility for practical application.

  16. Studies on optimization of neomycin fermentation liquid extraction process%新霉素发酵液提取工艺的优化研究

    Institute of Scientific and Technical Information of China (English)

    邱小明

    2013-01-01

    Because neomycin is sensitive to heat, the extraction rate of the existing extraction technology by applying the evaporation deamination and concentration is only 75%. The residual neomycin is discharged as wast ewater increasing pollutant discharge. Through the improved process using dynamic adsorption and elution of 732 resin in this study, the exchange capacity of neomycin is 142.9 mg per ml resin, the elution yield is higher than 96%, while reducing water consumption. The decoloration effect of LSA - 700 resin is better than 711 resin, whose neomycin recovery rate reaches 92.1% and pigment OD value is 0.38. Finally, the NH4OH in materials can be eff ectively removed by nanofiltration membrane of 200Da, the neomycin yield is higher than 99.4%, and the by-prod uct ammonia can be recycled. After the optimization, the neomycin extraction rate is 88.2%, higher than the existi ng level, showing this process can be well applied to neomycin extraction.%  新霉素对热较敏感,现有提取工艺主要采用蒸发脱氨和浓缩,提取率仅达75%,残余的新霉素以废水方式排放,增加了污染物的排放.本研究通过工艺改进经732树脂动态吸附与洗脱,新霉素的交换量为142.9 mg•ml-1树脂,洗脱收率高于96%,同时减少了工艺水的用量.用LSA—700树脂脱色效果优异711树脂,新霉素回收率达92.1%,色素OD值为0.38.最后通过200Da 分子量的纳滤膜组件处理可有效去除物料中的NH4OH,新霉素的收率高于99.4%,副产物氨水可回收利用.优化后的新霉素提取率达88.2%,高于工厂现有水平,说明本工艺能够很好地应用于新霉素的提取.

  17. Activation of Dormant Secondary Metabolite Production by Introducing Neomycin Resistance into the Deep-Sea Fungus, Aspergillus versicolor ZBY-3

    Directory of Open Access Journals (Sweden)

    Yuan Dong

    2014-07-01

    Full Text Available A new ultrasound-mediated approach has been developed to introduce neomycin-resistance to activate silent pathways for secondary metabolite production in a bio-inactive, deep-sea fungus, Aspergillus versicolor ZBY-3. Upon treatment of the ZBY-3 spores with a high concentration of neomycin by proper ultrasound irradiation, a total of 30 mutants were obtained by single colony isolation. The acquired resistance of the mutants to neomycin was confirmed by a resistance test. In contrast to the ZBY-3 strain, the EtOAc extracts of 22 of the 30 mutants inhibited the human cancer K562 cells, indicating that these mutants acquired a capability to produce antitumor metabolites. HPLC-photodiode array detector (PDAD-UV and HPLC-electron spray ionization (ESI-MS analyses of the EtOAc extracts of seven bioactive mutants and the ZBY-3 strain indicated that diverse secondary metabolites have been newly produced in the mutant extracts in contrast to the ZBY-3 extract. The followed isolation and characterization demonstrated that six metabolites, cyclo(d-Pro-d-Phe (1, cyclo(d-Tyr-d-Pro (2, phenethyl 5-oxo-l-prolinate (3, cyclo(l-Ile-l-Pro (4, cyclo(l-Leu-l-Pro (5 and 3β,5α,9α-trihydroxy-(22E,24R-ergosta-7,22-dien-6-one (6, were newly produced by the mutant u2n2h3-3 compared to the parent ZBY-3 strain. Compound 3 was a new compound; 2 was isolated from a natural source for the first time, and all of these compounds were also not yet found in the metabolites of other A. versicolor strains. Compounds 1–6 inhibited the K562 cells, with inhibition rates of 54.6% (1, 72.9% (2, 23.5% (3, 29.6% (4, 30.9% (5 and 51.1% (6 at 100 μg/mL, and inhibited also other human cancer HL-60, BGC-823 and HeLa cells, to some extent. The present study demonstrated the effectiveness of the ultrasound-mediated approach to activate silent metabolite production in fungi by introducing acquired resistance to aminoglycosides and its potential for discovering new compounds from silent

  18. Activation of dormant secondary metabolite production by introducing neomycin resistance into the deep-sea fungus, Aspergillus versicolor ZBY-3.

    Science.gov (United States)

    Dong, Yuan; Cui, Cheng-Bin; Li, Chang-Wei; Hua, Wei; Wu, Chang-Jing; Zhu, Tian-Jiao; Gu, Qian-Qun

    2014-07-29

    A new ultrasound-mediated approach has been developed to introduce neomycin-resistance to activate silent pathways for secondary metabolite production in a bio-inactive, deep-sea fungus, Aspergillus versicolor ZBY-3. Upon treatment of the ZBY-3 spores with a high concentration of neomycin by proper ultrasound irradiation, a total of 30 mutants were obtained by single colony isolation. The acquired resistance of the mutants to neomycin was confirmed by a resistance test. In contrast to the ZBY-3 strain, the EtOAc extracts of 22 of the 30 mutants inhibited the human cancer K562 cells, indicating that these mutants acquired a capability to produce antitumor metabolites. HPLC-photodiode array detector (PDAD)-UV and HPLC-electron spray ionization (ESI)-MS analyses of the EtOAc extracts of seven bioactive mutants and the ZBY-3 strain indicated that diverse secondary metabolites have been newly produced in the mutant extracts in contrast to the ZBY-3 extract. The followed isolation and characterization demonstrated that six metabolites, cyclo(D-Pro-D-Phe) (1), cyclo(D-Tyr-D-Pro) (2), phenethyl 5-oxo-L-prolinate (3), cyclo(L-Ile-L-Pro) (4), cyclo(L-Leu-L-Pro) (5) and 3β,5α,9α-trihydroxy-(22E,24R)-ergosta-7,22-dien-6-one (6), were newly produced by the mutant u2n2h3-3 compared to the parent ZBY-3 strain. Compound 3 was a new compound; 2 was isolated from a natural source for the first time, and all of these compounds were also not yet found in the metabolites of other A. versicolor strains. Compounds 1-6 inhibited the K562 cells, with inhibition rates of 54.6% (1), 72.9% (2), 23.5% (3), 29.6% (4), 30.9% (5) and 51.1% (6) at 100 μg/mL, and inhibited also other human cancer HL-60, BGC-823 and HeLa cells, to some extent. The present study demonstrated the effectiveness of the ultrasound-mediated approach to activate silent metabolite production in fungi by introducing acquired resistance to aminoglycosides and its potential for discovering new compounds from silent fungal

  19. Gold nanorod-covered kanamycin-loaded hollow SiO2 (HSKAurod) nanocapsules for drug delivery and photothermal therapy on bacteria

    Science.gov (United States)

    Hu, Bo; Zhang, Li-Pei; Chen, Xu-Wei; Wang, Jian-Hua

    2012-12-01

    A hybrid bactericidal material, gold nanorod-covered kanamycin-loaded hollow SiO2 (HSKAurod) nanocapsules, is constructed. The hybrid material combines the features of a chemical drug with photothermal physical sterilization which decreases the dosage of broad-spectrum antibiotic and the physical damage of biological systems. Hollow SiO2 nanocapsules are used as carriers for drug delivery. The nanocapsules load a model drug, kanamycin, and are covered with gold nanorods to avoid drug leakage and realize photothermal treatment. The sterilizing effect on the bacterial strain is investigated by incubating E. coli BL21 with the hybrid nanocapsules and irradiating under near-infrared light (NIR) for 20 min. A bactericidal effect, i.e., a sterilizing rate of 53.47%, is achieved for the HSKAurod nanocapsules under NIR irradiation, with respect to a net sum sterilizing rate of 34.49% for the individual components of the HSKAurod nanocapsules, e.g., carrier nanocapsules, chemical sterilization of kanamycin and physical sterilization due to the gold nanorods under NIR irradiation. It is demonstrated that the combination of chemical drug and physical sterilization results in an obvious synergistic effect and makes the sterilization more effective. This novel hybrid has great potential as an adjuvant therapeutic alternative material for sterilization or even for the control of disease.

  20. Study on the competitive reaction between bovine serum albumin and neomycin with ponceau S as fluorescence probe

    Energy Technology Data Exchange (ETDEWEB)

    Liu Baosheng, E-mail: lbs@hbu.edu.c [Key Laboratory of Medical Chemistry and Molecular Diagnosis, Ministry of Education, Center of Physics and Chemistry, Hebei University, Baoding 071002 (China); Xue Chunli; Wang Jing; Yang Chao; Zhao Fengli; Lv Yunkai [Key Laboratory of Medical Chemistry and Molecular Diagnosis, Ministry of Education, Center of Physics and Chemistry, Hebei University, Baoding 071002 (China)

    2010-11-15

    A competitive reaction exists between bovine serum albumin (BSA) and neomycin (NM) when ponceau S (PS) is chosen as fluorescent probe. This reaction was studied by fluorescence spectroscopy and UV-vis absorption spectroscopy. The static fluorescence quenching process between BSA and PS was confirmed and the binding constant, the number of binding sites and type of interaction forces between BSA and PS were obtained. It was observed that when NM was added into BSA-PS system, the relative fluorescence intensity of BSA was recovered gradually with increase in concentration of NM, which shows that there existed competitive reaction between BSA and NM. According to competitive reaction mechanism, the equilibrium concentration of PS, the binding constant and the type of interaction forces between PS and NM were obtained.

  1. Foscarnet Injection

    Science.gov (United States)

    ... Zovirax, Sitavig); aminoglycoside antibiotics such as amikacin, kanamycin, neomycin (Neo-Fradin), paromomycin, streptomycin, and tobramycin; amphotericin (Abelcet, Ambisome, Amphotec, Fungizone); cyclosporine ( ...

  2. Vancomycin Injection

    Science.gov (United States)

    ... Abelcet, Ambisome, Amphotec), bacitracin (Baciim); cisplatin, colistin, kanamycin, neomycin (Neo-Fradin), paromomycin, polymyxin B, streptomycin, and tobramycin. Your doctor may need to change ...

  3. Acyclovir

    Science.gov (United States)

    ... such as amikacin (Amikin), gentamicin (Garamycin), kanamycin (Kantrex), neomycin (Nes-RX, Neo-Fradin), paramomycin (Humatin), streptomycin, and tobramycin (Tobi, Nebcin); aspirin and other nonsteroidal ...

  4. Adefovir

    Science.gov (United States)

    ... such as amikacin (Amikin), gentamicin (Garamycin), kanamycin (Kantrex), neomycin (Neo-Rx, NeoFradin), paramomycin (Humatin), streptomycin, and tobramycin (Tobi, Nebcin); amphotericin B (Fungizone); aspirin ...

  5. Colistimethate Injection

    Science.gov (United States)

    ... Abelcet, Ambisome), capreomycin (Capastat), gentamicin (Gentak, Genoptic), kanamycin, neomycin (Neo-Fradin), paromomycin, polymyxin B, sodium citrate (in Bicitra), streptomycin, tobramycin (Tobi, Tobrex), or vancomycin (Vancocin). Your doctor ...

  6. Binding affinity and inhibitory potency of neomycin and streptomycin on the Tat peptide interaction with HIV-1 TAR RNA detected by on-line acoustic wave sensor.

    Science.gov (United States)

    Tassew, Nardos; Thompson, Michael

    2003-10-07

    The binding of two aminoglycoside antibiotics, neomycin and streptomycin, to a segment of the transactivation responsive region (TAR) RNA of the human immunodeficiency virus, and their inhibitory potency to disrupt the interaction of the RNA with a regulatory Tat protein-derived peptide, have been studied using a flow-through acoustic wave detector system. Binding affinity is directly correlated with the inhibitory potency of these molecules and the acoustic wave detection system shows that neomycin exhibits at least a ten-fold greater affinity for TAR RNA and that it is also a more potent inhibitor than streptomycin. These results are in agreement with previous studies. However, unlike the time-consuming batch-based assays, use of the flow-through format offers considerable potential for the rapid screening of the chemistry of relatively small-molecule-nucleic acid binding events.

  7. Synthesis of C-5, C-2' and C-4'-neomycin-conjugated triplex forming oligonucleotides and their affinity to DNA-duplexes.

    Science.gov (United States)

    Tähtinen, Ville; Granqvist, Lotta; Virta, Pasi

    2015-08-01

    Neomycin-conjugated homopyrimidine oligo 2'-deoxyribonucleotides have been synthesized on a solid phase and their potential as triplex forming oligonucleotides (TFOs) with DNA-duplexes has been studied. For the synthesis of the conjugates, C-5, C-2' and C-4'-tethered alkyne-modified nucleoside derivatives were used as an integral part of the standard automated oligonucleotide chain elongation. An azide-derived neomycin was then conjugated to the incorporated terminal alkynes by Cu(I)-catalyzed 1,3-dipolar cycloaddition (the click chemistry). Concentrated ammonia released the desired conjugates in acceptable purity and yields. The site of conjugation was expectedly important for the Hoogsteen-face recognition: C-5-conjugation showed a notable positive effect, whereas the influence of the C-2' and C-4'-modification remained marginal. In addition to conventional characterization methods (UV- and CD-spectroscopy), (19)F NMR spectroscopy was applied for the monitoring of triplex/duplex/single strand-conversions.

  8. Activation of the Silent Secondary Metabolite Production by Introducing Neomycin-Resistance in a Marine-Derived Penicillium purpurogenum G59

    Directory of Open Access Journals (Sweden)

    Chang-Jing Wu

    2015-04-01

    Full Text Available Introduction of neomycin-resistance into a marine-derived, wild-type Penicillium purpurogenum G59 resulted in activation of silent biosynthetic pathways for the secondary metabolite production. Upon treatment of G59 spores with neomycin and dimethyl sulfoxide (DMSO, a total of 56 mutants were obtained by single colony isolation. The acquired resistance of mutants to neomycin was testified by the resistance test. In contrast to the G59 strain, the EtOAc extracts of 28 mutants inhibited the human cancer K562 cells, indicating that the 28 mutants have acquired the capability to produce bioactive metabolites. HPLC-photodiode array detector (PDAD-UV and HPLC-electron spray ionization (ESI-MS analyses further indicated that diverse secondary metabolites have been newly produced in the bioactive mutant extracts. Followed isolation and characterization demonstrated that five bioactive secondary metabolites, curvularin (1, citrinin (2, penicitrinone A (3, erythro-23-O-methylneocyclocitrinol (4 and 22E-7α-methoxy-5α, 6α-epoxyergosta-8(14,22-dien-3β-ol (5, were newly produced by a mutant, 4-30, compared to the G59 strain. All 1–5 were also not yet found in the secondary metabolites of other wild type P. purpurogenum strains. Compounds 1–5 inhibited human cancer K562, HL-60, HeLa and BGC-823 cells to varying extents. Both present bioassays and chemical investigations demonstrated that the introduction of neomycin-resistance into the marine-derived fungal G59 strain could activate silent secondary metabolite production. The present work not only extended the previous DMSO-mediated method for introducing drug-resistance in fungi both in DMSO concentrations and antibiotics, but also additionally exemplified effectiveness of this method for activating silent fungal secondary metabolites. This method could be applied to other fungal isolates to elicit their metabolic potentials to investigate secondary metabolites from silent biosynthetic pathways.

  9. Activation of the silent secondary metabolite production by introducing neomycin-resistance in a marine-derived Penicillium purpurogenum G59.

    Science.gov (United States)

    Wu, Chang-Jing; Yi, Le; Cui, Cheng-Bin; Li, Chang-Wei; Wang, Nan; Han, Xiao

    2015-04-22

    Introduction of neomycin-resistance into a marine-derived, wild-type Penicillium purpurogenum G59 resulted in activation of silent biosynthetic pathways for the secondary metabolite production. Upon treatment of G59 spores with neomycin and dimethyl sulfoxide (DMSO), a total of 56 mutants were obtained by single colony isolation. The acquired resistance of mutants to neomycin was testified by the resistance test. In contrast to the G59 strain, the EtOAc extracts of 28 mutants inhibited the human cancer K562 cells, indicating that the 28 mutants have acquired the capability to produce bioactive metabolites. HPLC-photodiode array detector (PDAD)-UV and HPLC-electron spray ionization (ESI)-MS analyses further indicated that diverse secondary metabolites have been newly produced in the bioactive mutant extracts. Followed isolation and characterization demonstrated that five bioactive secondary metabolites, curvularin (1), citrinin (2), penicitrinone A (3), erythro-23-O-methylneocyclocitrinol (4) and 22E-7α-methoxy-5α, 6α-epoxyergosta-8(14),22-dien-3β-ol (5), were newly produced by a mutant, 4-30, compared to the G59 strain. All 1-5 were also not yet found in the secondary metabolites of other wild type P. purpurogenum strains. Compounds 1-5 inhibited human cancer K562, HL-60, HeLa and BGC-823 cells to varying extents. Both present bioassays and chemical investigations demonstrated that the introduction of neomycin-resistance into the marine-derived fungal G59 strain could activate silent secondary metabolite production. The present work not only extended the previous DMSO-mediated method for introducing drug-resistance in fungi both in DMSO concentrations and antibiotics, but also additionally exemplified effectiveness of this method for activating silent fungal secondary metabolites. This method could be applied to other fungal isolates to elicit their metabolic potentials to investigate secondary metabolites from silent biosynthetic pathways.

  10. Comparing amikacin and kanamycin-induced hearing loss in multidrug-resistant tuberculosis treatment under programmatic conditions in a Namibian retrospective cohort.

    Science.gov (United States)

    Sagwa, Evans L; Ruswa, Nunurai; Mavhunga, Farai; Rennie, Timothy; Leufkens, Hubert G M; Mantel-Teeuwisse, Aukje K

    2015-12-10

    Amikacin and kanamycin are mainly used for treating multidrug-resistant tuberculosis (MDR-TB), especially in developing countries where the burden of MDR-TB is highest. Their protracted use in MDR-TB treatment is known to cause dose-dependent irreversible hearing loss, requiring hearing aids, cochlear implants or rehabilitation. Therapeutic drug monitoring and regular audiological assessments may help to prevent or detect the onset of hearing loss, but these services are not always available or affordable in many developing countries. We aimed to compare the cumulative incidence of hearing loss among patients treated for MDR-TB with amikacin or kanamycin-based regimens, and to identify the most-at-risk patients, based on the real-life clinical practice experiences in Namibia. We conducted a retrospective cohort study of patients treated with amikacin or kanamycin-based regimens in four public sector MDR-TB treatment sites in Namibia between June 2004 and March 2014. Patients were audiologically assessed as part of clinical care. The study outcome was the occurrence of any hearing loss. Data were manually extracted from patients' treatment records. We compared proportions using the Chi-square test; applied stratified analysis and logistic regression to study the risk of hearing loss and to identify the most-at-risk patients through effect-modification analysis. A P-value < 0.05 was statistically significant. All 353 patients had normal baseline hearing, 46 % were HIV co-infected. Cumulative incidence of any hearing loss was 58 %, which was mostly bilateral (83 %), and mild (32 %), moderate (23 %), moderate-severe (16 %), severe (10 %), or profound (15 %). Patients using amikacin had a greater risk of developing the more severe forms of hearing loss than those using kanamycin (adjusted odds ratio (OR) = 4.0, 95 % CI: 1.5-10.8). Patients co-infected with HIV (OR = 3.4, 95 % CI: 1.1-10.6), males (OR = 4.5, 95 %1.5-13.4) and those with lower

  11. 新唑漱口液的质量控制%Quality Control of Neomycin and Metronidazole Gargarism

    Institute of Scientific and Technical Information of China (English)

    余晓霞; 刘春霞; 朱桦

    2014-01-01

    OBJECTIVE To establish a method for quality control of Neomycin and Metronidazole Gargarism.METHODS Neomycinsulphate and metronidazole of the gargarism were identified by chemical method ,ultraviolet spectrophotometry and thin-layer chromatography according to the methods in Chinese Pharmacopoeia 2010.The UV spectrophotometry was used to measure metronidazole concentration of the gargarism.RESULTS The characteristic identification reactions were positive of all main components.Determination of metronidazole yielded a linear range of 5.53~27.65μg· mL-1 with the regression equation of A =26.9627 C-0.0698 ,r=0.9999 ( n=7) ,the average re-covery of 100.04%and RSD of 1.01%( n=9).CONCLUSION The methods are simple ,rapid,sensitive and ac-curate.So it can be used for the quality control of Neomycin and Metronidazole Gargarism.%目的:建立新唑漱口液的质量控制方法。方法参照《中国药典》2010年版,采用化学法、紫外分光光度法以及薄层色谱法分别对新唑漱口液中主要成分甲硝唑和硫酸新霉素进行鉴别,采用紫外分光光度法测定甲硝唑的含量。结果新唑漱口液中的主要成分的鉴别反应均呈阳性;甲硝唑含量测定线性范围为5.53~27.65μg· mL 1,回归方程为A=26.9627C0.0698,r=0.9999(n=7),平均加样回收率为100.04%, RSD=1.01%(n=9)。结论该方法简单快速、灵敏准确,可用于新唑漱口液的质量控制。

  12. Development of ELISA for the detection of neomycin:preparation of anti-neomycin antibodies%新霉素酶联免疫检测方法的研究——新霉素抗体的制备

    Institute of Scientific and Technical Information of China (English)

    刘沙洲; 黄非; 王丽丽; 吕雪艳; 阮琨; 乔代蓉; 曹毅; 白林含

    2007-01-01

    以碳化二亚胺作为偶联剂,分别将新霉素(Neomycin,NEO)和牛血清白蛋白(Bovine serum albumin,BSA)、血蓝蛋白(KeyholeLimpet hemocyanin,KLH)进行偶联,得到偶联产物BSA-NEO和KLH-NEO.对BSA、KLH、NEO及其偶联产物BSA-NEO、KLH-NEO进行红外光谱分析,结果显示偶联产物的红外光谱中同时存在载休蛋白和NEO的特征吸收峰,初步证明偶联成功;将偶联产物BSA-NEO和KLH-NEO作为新霉素免疫抗原,分别免疫新西兰大白兔,成功获取新霉素抗血清,间接ELISA法测得抗血清效价可达1.28×105以上.

  13. The expression of NLRX1 in C57BL/6 mice cochlear hair cells: Possible relation to aging- and neomycin-induced deafness.

    Science.gov (United States)

    Yang, Qianqian; Sun, Gaoying; Cao, Zhixin; Yin, Haiyan; Qi, Qi; Wang, Jinghan; Liu, Wenwen; Bai, Xiaohui; Wang, Haibo; Li, Jianfeng

    2016-03-11

    Nucleotide-binding domain and leucine-rich-repeat-containing family member X1 (NLRX1) is a cytoplasmic pattern recognition receptor that is predominantly located in mitochondria, which is tightly related to mitochondrial damage, reactive oxygen species (ROS) production, inflammation and apoptosis. The present study was designed to explore whether NLRX1 expresses in C57BL/6 mice cochlear hair cells and, if so, to investigate the possible correlations between NLRX1 and hearing. The location and dynamic expression of NLRX1 were investigated by immunofluorescence, real-time PCR and Western blotting. Hearing thresholds of C57BL/6 mice were measured by auditory brainstem response (ABR). Moreover, the downstream inflammatory and apoptotic pathways regulated by NLRX1 were examined in age-related and neomycin-induced hair cell damage. Data showed that NLRX1 expressed in cytoplasm of C57BL/6 cochlear hair cells, especially in the cilia, which were essential for sound sensation. The expression of NLRX1 in hair cells increased as the mice grew up, and, decreased as they aged. Additionally, the activated apoptotic JNK pathway was detected in 9-month old mice with worse-hearing and 3-month old mice treated with neomycin. Overall, results indicate that NLRX1 may relate to hair cell maturity, hearing formation and maintenance, and promote hair cell apoptosis through JNK pathway induced by aging and neomycin.

  14. Determination of neomycin and oxytetracycline in the presence of their impurities in veterinary dosage forms by high-performance liquid chromatography/tandem mass spectrometry.

    Science.gov (United States)

    Vucićević-Prcetić, Katarina; Cservenák, Robert; Radulović, Niko

    2011-01-01

    Two HPLC/MS/MS methods, one for determination of neomycin sulfate and the other for determination of oxytetracycline hydrochloride in the presence of their impurities, were developed and validated. Separations were achieved with gradient elution on a C18 column. All components were ionized by positive-ion electrospray and detected by multiple reaction monitoring. Calibration curves were linear, with correlation coefficients >0.99. Precision of the methods was confirmed by RSD values of 0.34 and 0.71% for neomycin and oxytetracycline, respectively. Recovery values of 101.5 and 101.0%, respectively, indicated adequate accuracy. Analysis time for neomycin was 24 min, with the retention time of the main compound at 10.1 min; for oxytetracycline, the analysis time was 18 min, with the main peak at 9.95 min. Longer retention times than expected were a consequence of the necessity of chromatographic separation of isomers with the same ion transition. All impurities defined in the pharmacopoeias were determined and their identities confirmed. The methods were tested for QC of veterinary dosage forms (commercial powders and injections containing these antibiotics).

  15. The neomycin biosynthetic gene cluster of Streptomyces fradiae NCIMB 8233: genetic and biochemical evidence for the roles of two glycosyltransferases and a deacetylase.

    Science.gov (United States)

    Fan, Qingzhi; Huang, Fanglu; Leadlay, Peter F; Spencer, Jonathan B

    2008-09-21

    An efficient protocol has been developed for the genetic manipulation of Streptomyces fradiae NCIMB 8233, which produces the 2-deoxystreptamine (2-DOS)-containing aminoglycoside antibiotic neomycin. This has allowed the in vivo analysis of the respective roles of the glycosyltransferases Neo8 and Neo15, and of the deacetylase Neo16 in neomycin biosynthesis. Specific deletion of each of the neo8, neo15 and neo16 genes confirmed that they are all essential for neomycin biosynthesis. The pattern of metabolites produced by feeding putative pathway intermediates to these mutants provided unambiguous support for a scheme in which Neo8 and Neo15, whose three-dimensional structures are predicted to be highly similar, have distinct roles: Neo8 catalyses transfer of N-acetylglucosamine to 2-DOS early in the pathway, while Neo15 catalyses transfer of the same aminosugar to ribostamycin later in the pathway. The in vitro substrate specificity of Neo15, purified from recombinant Escherichia coli, was fully consistent with these findings. The in vitro activity of Neo16, the only deacetylase so far recognised in the neo gene cluster, showed that it is capable of acting in tandem with both Neo8 and Neo15 as previously proposed. However, the deacetylation of N-acetylglucosaminylribostamycin was still observed in a strain deleted of the neo16 gene and fed with suitable pathway precursors, providing evidence for the existence of a second enzyme in S. fradiae with this activity.

  16. Mechanistic study on the nuclear modifier gene MSS1 mutation suppressing neomycin sensitivity of the mitochondrial 15S rRNA C1477G mutation in Saccharomyces cerevisiae.

    Science.gov (United States)

    Zhou, Qiyin; Wang, Wei; He, Xiangyu; Zhu, Xiaoyu; Shen, Yaoyao; Yu, Zhe; Wang, Xuexiang; Qi, Xuchen; Zhang, Xuan; Fan, Mingjie; Dai, Yu; Yang, Shuxu; Yan, Qingfeng

    2014-01-01

    The phenotypic manifestation of mitochondrial DNA (mtDNA) mutations can be modulated by nuclear genes and environmental factors. However, neither the interaction among these factors nor their underlying mechanisms are well understood. The yeast Saccharomyces cerevisiae mtDNA 15S rRNA C1477G mutation (PR) corresponds to the human 12S rRNA A1555G mutation. Here we report that a nuclear modifier gene mss1 mutation suppresses the neomycin-sensitivity phenotype of a yeast C1477G mutant in fermentable YPD medium. Functional assays show that the mitochondrial function of the yeast C1477G mutant was impaired severely in YPD medium with neomycin. Moreover, the mss1 mutation led to a significant increase in the steady-state level of HAP5 (heme activated protein), which greatly up-regulated the expression of glycolytic transcription factors RAP1, GCR1, and GCR2 and thus stimulated glycolysis. Furthermore, the high expression of the key glycolytic enzyme genes HXK2, PFK1 and PYK1 indicated that enhanced glycolysis not only compensated for the ATP reduction from oxidative phosphorylation (OXPHOS) in mitochondria, but also ensured the growth of the mss1(PR) mutant in YPD medium with neomycin. This study advances our understanding of the phenotypic manifestation of mtDNA mutations.

  17. c-Myb knockdown increases the neomycin-induced damage to hair-cell-like HEI-OC1 cells in vitro.

    Science.gov (United States)

    Yu, Xiaoyu; Liu, Wenwen; Fan, Zhaomin; Qian, Fuping; Zhang, Daogong; Han, Yuechen; Xu, Lei; Sun, Gaoying; Qi, Jieyu; Zhang, Shasha; Tang, Mingliang; Li, Jianfeng; Chai, Renjie; Wang, Haibo

    2017-01-23

    c-Myb is a transcription factor that plays a key role in cell proliferation, differentiation, and apoptosis. It has been reported that c-Myb is expressed within the chicken otic placode, but whether c-Myb exists in the mammalian cochlea, and how it exerts its effects, has not been explored yet. Here, we investigated the expression of c-Myb in the postnatal mouse cochlea and HEI-OC1 cells and found that c-Myb was expressed in the hair cells (HCs) of mouse cochlea as well as in cultured HEI-OC1 cells. Next, we demonstrated that c-Myb expression was decreased in response to neomycin treatment in both cochlear HCs and HEI-OC1 cells, suggesting an otoprotective role for c-Myb. We then knocked down c-Myb expression with shRNA transfection in HEI-OC1 cells and found that c-Myb knockdown decreased cell viability, increased expression of pro-apoptotic factors, and enhanced cell apoptosis after neomycin insult. Mechanistic studies revealed that c-Myb knockdown increased cellular levels of reactive oxygen species and decreased Bcl-2 expression, both of which are likely to be responsible for the increased sensitivity of c-Myb knockdown cells to neomycin. This study provides evidence that c-Myb might serve as a new target for the prevention of aminoglycoside-induced HC loss.

  18. Development of enzyme-linked immunosorbent assay (ELISA) for the detection of neomycin residues in pig muscle, chicken muscle, egg, fish, milk and kidney.

    Science.gov (United States)

    Wang, S; Xu, B; Zhang, Y; He, J X

    2009-05-01

    A colorimetric competitive direct enzyme-linked immunosorbent assay (ELISA) method was developed using polyclonal antibody to determine neomycin residues in food of animal origin. No cross-reactivity of the antibody was observed with other aminoglycosides. The limit of detection of the method was 0.1μg/kg. A simple and efficient sample extraction method was established with recoveries of neomycin ranged from 75% to 105%. The detection limits were 5μg/kg(l) in pig muscle, chicken muscle, fish and milk, 10μg/kg in kidney and 20μg/kg in egg, respectively. Chemiluminescence assay was developed for detecting neomycin residues in pig muscle and chicken muscle. The limit of detection of the method was 0.015μg/kg, and the detection limits were 1.5μg/kg in pig muscle and 6μg/kg in chicken muscle. The ELISA tests were validated by HPLC, and the results showed a good correlation (r(2)) which was greater than 0.9.

  19. Heterologous transformation of Agrocybe aegerita with a bacterial neomycin-resistance gene fused to a fungal promoter-like DNA sequence.

    Science.gov (United States)

    Noël, T; Simoneau, P; Labarère, J

    1995-06-01

    DNA sequences of the basidiomycete Agrocybe aegerita were cloned in E. coli based on their ability to drive the expression of the bacterial promoterless tetracycline (Tc)-resistance gene. A 0.48% frequency of the cloned sequences promoted antibiotic-resistance. The sequence conferring the highest Tc resistance (40 μg/ml) was selected to drive the expression in E. coli of two other promoterless genes encoding chloramphenicol and neomycin resistance. One of the derivative vectors, pN13-A2, carrying a chimeric neomycin-resistance gene, was used to transform an A. aegerita neomycin-sensitive strain by protoplast electroporation. Transformation frequencies ranged from 1 to 2.8 transformants per μg of DNA per 10(3) viable cells, in a relatively high background of spontaneous-resistant colonies (2% of the surviving protoplasts). Molecular analyses showed that transformation had occurred by the integration of pN13-A2 sequences, either ectopically or at the resident locus carrying the A. aegerita promoter-like sequence, with probable molecular rearrangements. The nucleotide sequence of the promoter-like fragment revealed the presence of a CT motif that is known to be involved in a promoter function in some highly expressed genes of filamentous fungi.

  20. Reduced TRMU expression increases the sensitivity of hair-cell-like HEI-OC-1 cells to neomycin damage in vitro

    Science.gov (United States)

    He, Zuhong; Sun, Shan; Waqas, Muhammad; Zhang, Xiaoli; Qian, Fuping; Cheng, Cheng; Zhang, Mingshu; Zhang, Shasha; Wang, Yongming; Tang, Mingliang; Li, Huawei; Chai, Renjie

    2016-01-01

    Aminoglycosides are ototoxic to the cochlear hair cells, and mitochondrial dysfunction is one of the major mechanisms behind ototoxic drug-induced hair cell death. TRMU (tRNA 5-methylaminomethyl-2-thiouridylate methyltransferase) is a mitochondrial protein that participates in mitochondrial tRNA modifications, but the role of TRMU in aminoglycoside-induced ototoxicity remains to be elucidated. In this study, we took advantage of the HEI-OC-1 cell line to investigate the role of TRMU in aminoglycoside-induced cell death. We found that TRMU is expressed in both hair cells and HEI-OC-1 cells, and its expression is significantly decreased after 24 h neomycin treatment. We then downregulated TRMU expression with siRNA and found that cell death and apoptosis were significantly increased after neomycin injury. Furthermore, when we down-regulated TRMU expression, we observed significantly increased mitochondrial dysfunction and increased levels of reactive oxygen species (ROS) after neomycin injury, suggesting that TRMU regulates mitochondrial function and ROS levels. Lastly, the antioxidant N-acetylcysteine rescued the mitochondrial dysfunction and cell apoptosis that was induced by TRMU downregulation, suggesting that ROS accumulation contributed to the increased aminoglycosides sensitivity of HEI-OC-1 cells after TRMU downregulation. This study provides evidence that TRMU might be a new therapeutic target for the prevention of aminoglycoside-induced hair cell death. PMID:27405449

  1. Gene-targeted embryonic stem cells: real-time PCR assay for estimation of the number of neomycin selection cassettes

    Directory of Open Access Journals (Sweden)

    Mancini Cecilia

    2011-10-01

    Full Text Available Abstract In the preparation of transgenic murine ES cells it is important to verify the construct has a single insertion, because an ectopic neomycin phosphortransferase positive selection cassette (NEO may cause a position effect. During a recent work, where a knockin SCA28 mouse was prepared, we developed two assays based on Real-Time PCR using both SYBR Green and specific minor groove binder (MGB probes to evaluate the copies of NEO using the comparative delta-delta Ct method versus the Rpp30 reference gene. We compared the results from Southern blot, routinely used to quantify NEO copies, with the two Real-Time PCR assays. Twenty-two clones containing the single NEO copy showed values of 0.98 ± 0.24 (mean ± 2 S.D., and were clearly distinguishable from clones with two or more NEO copies. This method was found to be useful, easy, sensitive and fast and could substitute for the widely used, but laborious Southern blot method.

  2. Cotransfection of Pax2 and Math1 promote in situ cochlear hair cell regeneration after neomycin insult.

    Science.gov (United States)

    Chen, Yan; Yu, Huiqian; Zhang, Yanping; Li, Wen; Lu, Na; Ni, Wenli; He, Yingzi; Li, Jin; Sun, Shan; Wang, Zhengmin; Li, Huawei

    2013-10-21

    The ideal strategy for hair cell regeneration is promoting residual supporting cell proliferation followed by induction of hair cell differentiation. In this study, cultured neonatal mouse organs of Corti were treated with neomycin to eliminate hair cells followed by incubation with recombined adenovirus expressing Pax2 and/or Math1. Results showed that overexpression of Pax2 significantly promoted proliferation of supporting cells. The number of BrdU⁺/myosin VIIA⁺ cells increased significantly in hair cell pre-existing region two weeks after adenovirus infection in Ad-Pax2-IRES-Math1 group compared to Ad-Pax2 and Ad-Math1 groups. This indicated that cotransfection of Pax2 and Math1 induced supporting cells to proliferate and differentiate into hair cells in situ. Most new hair cells were labeled by FM1-43 suggesting they acquired certain function. The results also suggest that inducing proliferating cells rather than quiescent cells to differentiate into hair cells by forced expression of Math1 is feasible for mammalian hair cell regeneration.

  3. Erythromycin versus neomycin in the treatment of hepatic encephalopathy in cirrhosis: a randomized double-blind study

    Directory of Open Access Journals (Sweden)

    Romeiro Fernando Gomes

    2013-01-01

    Full Text Available Abstract Background Hepatic encephalopathy (HE is a severe complication in patients with hepatic cirrhosis, which causes numerous hospital admissions and deaths. Antibiotics are the best options in HE treatment, but head-to-head comparisons between these drugs are scarce. Erythromycin combines the antimicrobial effect and prokinetic properties in the same drug, but it has never been used in HE treatment. Our aim was to evaluate the efficacy of erythromycin as an HE treatment. Methods We achieved a randomized controlled trial of adult patients with HE and hepatic cirrhosis admitted in our hospital. After randomization, the subjects received either erythromycin 250 mg or neomycin 1 g orally QID until hospital discharge or prescription of another antibiotic. All subjects were blindly evaluated every day towards quantifying clinical, neuropsychometric, hepatic and renal exams. Statistical analysis was employed to compare the groups and correlate the variables with hospitalization duration. Results 30 patients were evaluated (15 treated with each drug. At hospital admission, the groups were homogeneous, but the erythromycin group subjects achieved a shorter hospitalization stay (p = 0.032 and a more expressive reduction in alanine aminotranspherase levels (p = 0.026. Hospitalization duration was positively correlated with C reactive protein levels measured previous to (p = 0.015 and after treatment (p = 0.01. Conclusions In the sample evaluated erythromycin was associated with significant reductions in hospital stay and in alanine aminotranspherase values. Hospitalization time was positive correlated with C reactive protein levels measured before and after the treatments.

  4. Research progress of acute kanamycin sulfate-induced deafness in guinea pig%豚鼠硫酸卡那霉素急性致聋研究进展

    Institute of Scientific and Technical Information of China (English)

    殷泽登

    2012-01-01

    To present a summary of current knowledge regarding acute kanamycin sulfate-induced deafness in guinea pig, by reviewing the published literature. Animal model of acute deafness induced by a single dose of kanamycin sulfate in combination with ethacrynic acid or furosemide in guinea pig was usually used to investigate the mechanism of cochlear cell degeneration. There were different time sequences of cell degeneration of spiral ganglion cell and hair cell in different studies. The findings may result from different doses, order of two drugs administration or rime point chosen. There remains scope for further research in chronic kanamycin-induced deafness, which more replicates the type of exposure to people than acute deafness.

  5. β-lactams and florfenicol antibiotics remain bioactive in soils while ciprofloxacin, neomycin, and tetracycline are neutralized.

    Science.gov (United States)

    Subbiah, Murugan; Mitchell, Shannon M; Ullman, Jeffrey L; Call, Douglas R

    2011-10-01

    It is generally assumed that antibiotic residues in soils select for antibiotic-resistant bacteria. This assumption was tested by separately adding 10 different antibiotics (≥200 ppm) to three soil-water slurries (silt-loam, sand-loam, and sand; 20% soil [wt/vol]) and incubating mixtures for 24 h at room temperature. The antibiotic activity of the resultant supernatant was assessed by culturing a sensitive Escherichia coli strain in the filter-sterilized supernatant augmented with Luria-Bertani broth. We found striking differences in the abilities of supernatants to suppress growth of the indicator E. coli. Ampicillin, cephalothin, cefoxitin, ceftiofur, and florfenicol supernatants completely inhibited growth while bacterial growth was uninhibited in the presence of neomycin, tetracycline, and ciprofloxacin supernatants. High-performance liquid chromatography (HPLC) analysis demonstrated that cefoxitin and florfenicol were almost completely retained in the supernatants, whereas tetracycline and ciprofloxacin were mostly removed. Antibiotic dissipation in soil, presumably dominated by adsorption mechanisms, was sufficient to neutralize 200 ppm of tetracycline; this concentration is considerably higher than reported contamination levels. Soil pellets from the tetracycline slurries were resuspended in a minimal volume of medium to maximize the interaction between bacteria and soil particles, but sensitive bacteria were still unaffected by tetracycline (P = 0.6). Thus, residual antibiotics in soil do not necessarily exert a selective pressure, and the degree to which the pharmaceutical remains bioactive depends on the antibiotic. Efforts to control antibiotic contamination would be better directed toward compounds that retain biological activity in soils (e.g., cephalosporins and florfenicol) because these are the antibiotics that could exert a selective pressure in the environment.

  6. Conjugation of a Ru(II) arene complex to neomycin or to guanidinoneomycin leads to compounds with differential cytotoxicities and accumulation between cancer and normal cells.

    Science.gov (United States)

    Grau-Campistany, Ariadna; Massaguer, Anna; Carrion-Salip, Dolors; Barragán, Flavia; Artigas, Gerard; López-Senín, Paula; Moreno, Virtudes; Marchán, Vicente

    2013-05-01

    A straightforward methodology for the synthesis of conjugates between a cytotoxic organometallic ruthenium(II) complex and amino- and guanidinoglycosides, as potential RNA-targeted anticancer compounds, is described. Under microwave irradiation, the imidazole ligand incorporated on the aminoglycoside moiety (neamine or neomycin) was found to replace one triphenylphosphine ligand from the ruthenium precursor [(η(6)-p-cym)RuCl(PPh3)2](+), allowing the assembly of the target conjugates. The guanidinylated analogue was easily prepared from the neomycin-ruthenium conjugate by reaction with N,N'-di-Boc-N″-triflylguanidine, a powerful guanidinylating reagent that was compatible with the integrity of the metal complex. All conjugates were purified by semipreparative high-performance liquid chromatography (HPLC) and characterized by electrospray ionization (ESI) and matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) and NMR spectroscopy. The cytotoxicity of the compounds was tested in MCF-7 (breast) and DU-145 (prostate) human cancer cells, as well as in the normal HEK293 (Human Embryonic Kidney) cell line, revealing a dependence on the nature of the glycoside moiety and the type of cell (cancer or healthy). Indeed, the neomycin-ruthenium conjugate (2) displayed moderate antiproliferative activity in both cancer cell lines (IC50 ≈ 80 μM), whereas the neamine conjugate (4) was inactive (IC50 ≈ 200 μM). However, the guanidinylated analogue of the neomycin-ruthenium conjugate (3) required much lower concentrations than the parent conjugate for equal effect (IC50 = 7.17 μM in DU-145 and IC50 = 11.33 μM in MCF-7). Although the same ranking in antiproliferative activity was found in the nontumorigenic cell line (3 ≫ 2 > 4), IC50 values indicate that aminoglycoside-containing conjugates are about 2-fold more cytotoxic in normal cells (e.g., IC50 = 49.4 μM for 2) than in cancer cells, whereas an opposite tendency was found

  7. Bax, Bcl2, and p53 differentially regulate neomycin- and gentamicin-induced hair cell death in the zebrafish lateral line.

    Science.gov (United States)

    Coffin, Allison B; Rubel, Edwin W; Raible, David W

    2013-10-01

    Sensorineural hearing loss is a normal consequence of aging and results from a variety of extrinsic challenges such as excessive noise exposure and certain therapeutic drugs, including the aminoglycoside antibiotics. The proximal cause of hearing loss is often death of inner ear hair cells. The signaling pathways necessary for hair cell death are not fully understood and may be specific for each type of insult. In the lateral line, the closely related aminoglycoside antibiotics neomycin and gentamicin appear to kill hair cells by activating a partially overlapping suite of cell death pathways. The lateral line is a system of hair cell-containing sense organs found on the head and body of aquatic vertebrates. In the present study, we use a combination of pharmacologic and genetic manipulations to assess the contributions of p53, Bax, and Bcl2 in the death of zebrafish lateral line hair cells. Bax inhibition significantly protects hair cells from neomycin but not from gentamicin toxicity. Conversely, transgenic overexpression of Bcl2 attenuates hair cell death due to gentamicin but not neomycin, suggesting a complex interplay of pro-death and pro-survival proteins in drug-treated hair cells. p53 inhibition protects hair cells from damage due to either aminoglycoside, with more robust protection seen against gentamicin. Further experiments evaluating p53 suggest that inhibition of mitochondrial-specific p53 activity confers significant hair cell protection from either aminoglycoside. These results suggest a role for mitochondrial p53 activity in promoting hair cell death due to aminoglycosides, likely upstream of Bax and Bcl2.

  8. The field trials of neomycin against Erwinia carotovora subsp, carotovora%新霉素对大白菜软腐病的田间药效试验研究

    Institute of Scientific and Technical Information of China (English)

    王东磊; 达娃; 崔文华; 陈龙; 侯太平

    2011-01-01

    The stability of neomycin was tested by physical and chemical factors to study the antibacterial activity. Field trials of neomycin against Erwinia carotovora subsp. Carotovora was conducted to study neomycin control effect. Physical and chemical factors test showed that neomycin is very stable, it could resist strong acid, alkali, high temperature and UV light. Besides neomycin has many advantages in producing, storing, applying. The results of field trial show that neomycin has good control efficiency a-gainst Erwinia carotovora subsp. Carotovorat. The control effects of neomycin with 200 mg/L,100 mg/ L、50 mg/L were 84.1%,72. 83% and 63. 2% respectively. The control effect of streptomycin with 200 mg/L was 60. 24%. The test results show that neomycin has a promising prospect in pesticide industry. All these advantages make it a very good candidate for pestcide.%本研究通过各种理化因子对新霉素活性的影响考察新霉素的稳定性,进一步通过田间小区试验方法考察新霉素对大白菜软腐病的田间实际防治效果.理化因子试验结果表明,新霉素具有良好的稳定性,能够耐酸碱,耐高温、耐紫外线的直射,具有生产、储藏和应用上的优势.田间小区试验结果表明,新霉素200 mg/L、100 mg/L、50 mg/L的防治效果均优于常用药剂72%农用硫酸链霉素.新霉素200 mg/L、100 mg/L、50 mg/L的防治效果为84.1%、72.83%、63.2%.72%农用硫酸链霉素50mg/L的防治效果为60.24%.两组试验显示了新霉素具有良好的应用前景,为把新霉素开发为一种新型的抗生素类农药奠定了基础.

  9. Comparative biorelease study of fluticasone in combination with antibacterial (Neomycin and or antifungal (coltrimazol, miconazole agents by histamine percutaneous reaction method in healthy volunteers

    Directory of Open Access Journals (Sweden)

    Shahani S

    1997-01-01

    Full Text Available Fluticasone propionate is a novel, potent and topically active synthetic corticosteroid preparation with a much reduced potential, in relation to its anti-inflammatory potency, for unwanted systemic side effects. It is indicated for the treatment of eczema, dermatitis etc. The objective of the present study was to evaluate and compare the biorelease of fluticassone with placebo (base formulation; its combination with antifungal (miconazole, clotrimazole and / or antibacterial agents based on the attenuation of histamine induced wheal and flare reaction and flare intensity (on visual analouge scale by McNemar test. In the present study, fluticasone alone and in combination with clotrimazole, miconazole and neomycin significantly reduced the wheal and flare response of histamine prick test. The flare intensity response was also significantly inhibited by these treatments. Furthermore, there was no difference in the anti-inflammatory activity of various treatment groups. It may, therefore, be concluded that antibacterial (neomycin and / or antifungal (miconazole, clotrimazole agents in combination with steroid (fluticasone do not alter the pharmacodynamic response of the latter.

  10. Hydrocortisone, Neomycin, and Polymyxin

    Science.gov (United States)

    ... procedure easier, have someone else instill the drops): Wash your hands thoroughly with soap and warm water. Gently clean ... and tighten the cap or dropper right away. Wash your hands to remove any medication. To instill the eye ...

  11. Development of an Immunochromatographic Strip Test for Detection of Neomycin%新霉素免疫膜层析检测方法研究

    Institute of Scientific and Technical Information of China (English)

    王爱萍; 李发弟; 胡骁飞; 史兆国; 周景明; 杜晓明; 王林林; 禹乐乐; 刘明阳

    2011-01-01

    [Objective]The objective of this experiment is to develop an immunochromatographic strip for rapid detection of neomycin (NEO).[Method]The monoclonal antibodies (mAb) against neomycin were generated and labeled with colloidal gold particles.Then the immunochromatographic strips for detection of NEO were developed and identified.[Result]Ka of NEO monoclonal antibody was 3.75× 1010 mol· L-1.The detection limit of the strip was 50 ng·mL-1 by naked eyes.The cross-reactivity with other antibiotics, such as gentamicin, streptomycin, tobramycin, sisomicin, ractopamine, salbutamal, tylosin, chloramphenic, was not detected.In parallel experiments, the test strip gave the same results as HPLC-MS-MS did.[Conclusion]An immunochromatographic strip test method was established and the strip can be used to detect NEO residues rapidly with high specificity, sensitivity and simplicity in different samples.%[目的]研制新霉素(neomycin,NEO)免疫膜层析快速检测试纸.[方法]建立分泌抗新霉素单克隆抗体(NEO mAb)杂交瘤细胞株,用胶体金免疫膜层析试验(GICA)制备NEO残留快速检测试纸(NEO-Strip),并对其性能进行鉴定.[结果]抗新霉素单克隆抗体的亲和力常数为3.75×1010mol·L-1;制备的NEO-Strip目测限为50ng·mL-1;试纸可特异性检测NEO,与庆大霉素、链霉素、妥布霉素、紫苏霉素、莱克多巴胺、沙丁胺醇、泰乐菌素和氯霉素等其它药物均无交叉反应性;就定性检测而言,其结果与高效液相色谱串联质谱法(HPLC-MS-MS)完全一致;不同的基质液对试纸的敏感度影响甚微.[结论]建立了特异的NEO免疫层析检测方法.

  12. Binding of PLCδ1PH-GFP to Ptdlns(4,5)P2 prevents inhibition of phospholipase C-mediated hydrolysis of Ptdlns(4,5)P2 by neomycin

    Institute of Scientific and Technical Information of China (English)

    Chuan WANG; Xiao-na DU; Qing-zhong JIA; Hai-lin ZHANG

    2005-01-01

    Aim: To investigate the effects of the pleckstrin homology (PH) domain of phospholipase Cδ1 (PLCδ1PH) on inhibition of phospholipase C (PLC)-mediated hydrolysis of phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] by neomycin.Methods: A fusion construct of green fluorescent protein (GFP) and PLCδ1PH (PLCδ1PH-GFP), which is known to bind Ptdlns(4,5)P2 specifically, together with laser-scanning confocal microscopy, was used to trace PtdIns(4,5)P2 translocation.Results: Stimulation of the type 1 muscarinic receptor and the bradykinin 2 receptor induced a reversible PLCδ1PH-GFP translocation from the membrane to the cytosol in COS-7 cells. PLC inhibitor U73122 blocked the translocation.Wortmannin, a known PtdIns kinase inhibitor, did not affect the translocation induced by ACh, but blocked recovery after translocation, indicating that PtdIns(4,5)P2 hydrolysis occurs through receptor-mediated PLC activation.Neomycin, a commonly used phospholipase C blocker, failed to block the receptor-induced PLCδ1PH-GFP translocation, indicating that neomycin is unable to block PLC-mediated PtdIns(4,5)P2 hydrolysis. However, in the absence of PLCδ1PH-GFP expression, neomycin abolished the receptor-induced hydrolysis of PtdIns(4,5)P2 by PLC. Conclusion: Although PLCδ1PH and neomycin bind to PtdIns(4,5)P2 in a similar way, they have distinct effects on receptor-mediated activation of PLC and PtdIns(4,5)P2 hydrolysis.

  13. Neomycin damage and regeneration of hair cells in both mechanoreceptor and electroreceptor lateral line organs of the larval Siberian sturgeon (Acipenser baerii).

    Science.gov (United States)

    Fan, Chunxin; Zou, Sha; Wang, Jian; Zhang, Bo; Song, Jiakun

    2016-05-01

    The lateral line found in some amphibians and fishes has two distinctive classes of sensory organs: mechanoreceptors (neuromasts) and electroreceptors (ampullary organs). Hair cells in neuromasts can be damaged by aminoglycoside antibiotics and they will regenerate rapidly afterward. Aminoglycoside sensitivity and the capacity for regeneration have not been investigated in ampullary organs. We treated Siberian sturgeon (Acipenser baerii) larvae with neomycin and observed loss and regeneration of sensory hair cells in both organs by labeling with DASPEI and scanning electron microscopy (SEM). The numbers of sensory hair cells in both organs were reduced to the lowest levels at 6 hours posttreatment (hpt). New sensory hair cells began to appear at 12 hpt and were regenerated completely in 7 days. To reveal the possible mechanism for ampullary hair cell regeneration, we analyzed cell proliferation and the expression of neural placodal gene eya1 during regeneration. Both cell proliferation and eya1 expression were concentrated in peripheral mantle cells and both increased to the highest level at 12 hpt, which is consistent with the time course for regeneration of the ampullary hair cells. Furthermore, we used Texas Red-conjugated gentamicin in an uptake assay following pretreatment with a cation channel blocker (amiloride) and found that entry of the antibiotic was suppressed in both organs. Together, our results indicate that ampullary hair cells in Siberian sturgeon larvae can be damaged by neomycin exposure and they can regenerate rapidly. We suggest that the mechanisms for aminoglycoside uptake and hair cell regeneration are conserved for mechanoreceptors and electroreceptors. J. Comp. Neurol. 524:1443-1456, 2016. © 2015 Wiley Periodicals, Inc.

  14. Resistances Level of Indian Diploid and Tetraploid Cotton Cultivarsagainst Plant Selection Marker Kanamycin and Direct Shoot Organogenesis from Shoot Tip Culture%印度棉花二倍体和四倍体品种对卡那霉素的抗性水平及茎尖培养直接器官发生法诱导成苗

    Institute of Scientific and Technical Information of China (English)

    G. BALASUBRMANI; J. Amudha, C. D. MAYEE

    2002-01-01

    @@ Selection of transformed tissues in the antibiotic medium is an important step for developing transgenic plants. The toxic level of G.arboreum and G. hirsutum cotton cuhivars of Indian origin were tested against kanamycin using different concentrations.

  15. Detection of kanamycin and gentamicin residues in animal-derived food using IgY antibody based ic-ELISA and FPIA.

    Science.gov (United States)

    Li, Cui; Zhang, Yaoyao; Eremin, Sergei A; Yakup, Omar; Yao, Gang; Zhang, Xiaoying

    2017-07-15

    Our aim in this study is to show that IgY antibody based immunoassays could be used to detect antibiotic residues in animal-derived food. Briefly, full antigens of gentamicin (Gent) and kanamycin (Kana) were used to immunize the laying chickens to prepare IgY antibodies. Then, these antibodies were evaluated by FPIA and ic-ELISA to detect Gent/Kana in animal-derived samples. The IC50 of FPIA and ic-ELISA based anti-Gent IgY were 7.70±0.6μg/mL and 0.32±0.06μg/mL, respectively. The IC50 of FPIA and ic-ELISA based anti-Kana IgY were 7.97±0.9μg/mL and 0.15±0.01μg/mL. The limits of detection (LOD, IC10) for FPIA based anti-Gent/Kana IgY were 0.17 and 0.007μg/mL, respectively. The LOD for ic-ELISA were both 0.001μg/mL. These results indicated that the ic-ELISA might more suitable for antibiotic residues detection than FPIA.

  16. Kanamycin Sulphate Loaded PLGA-Vitamin-E-TPGS Long Circulating Nanoparticles Using Combined Coating of PEG and Water-Soluble Chitosan

    Science.gov (United States)

    Mustafa, Sanaul

    2017-01-01

    Kanamycin sulphate (KS) is a Mycobacterium tuberculosis protein synthesis inhibitor. Due to its intense hydrophilicity, KS is cleared from the body within 8 h. KS has a very short plasma half-life (2.5 h). KS is used in high concentrations to reach the therapeutic levels in plasma, which results in serious nephrotoxicity/ototoxicity. To overcome aforementioned limitations, the current study aimed to develop KS loaded PLGA-Vitamin-E-TPGS nanoparticles (KS-PLGA-TPGS NPs), to act as an efficient carrier for controlled delivery of KS. To achieve a substantial extension in blood circulation, a combined design, affixation of polyethylene glycol (PEG) to KS-PLGA-TPGS NPs and adsorption of water-soluble chitosan (WSC) (cationic deacetylated chitin) to particle surface, was raised for surface modification of NPs. Surface modified NPs (KS-PEG-WSC NPs) were prepared to provide controlled delivery and circulate in the bloodstream for an extended period of time, thus minimizing dosing frequency. In vivo pharmacokinetics and in vivo biodistribution following intramuscular administration were investigated. NPs surface charge was close to neutral +3.61 mV and significantly affected by the WSC coating. KS-PEG-WSC NPs presented striking prolongation in blood circulation, reduced protein binding, and long drew-out the blood circulation half-life with resultant reduced kidney sequestration vis-à-vis KS-PLGA-TPGS NPs. The studies, therefore, indicate the successful formulation development of KS-PEG-WSC NPs with reduced frequency of dosing of KS indicating low incidence of nephrotoxicity/ototoxicity.

  17. Kanamycin Sulphate Loaded PLGA-Vitamin-E-TPGS Long Circulating Nanoparticles Using Combined Coating of PEG and Water-Soluble Chitosan

    Directory of Open Access Journals (Sweden)

    Sanaul Mustafa

    2017-01-01

    Full Text Available Kanamycin sulphate (KS is a Mycobacterium tuberculosis protein synthesis inhibitor. Due to its intense hydrophilicity, KS is cleared from the body within 8 h. KS has a very short plasma half-life (2.5 h. KS is used in high concentrations to reach the therapeutic levels in plasma, which results in serious nephrotoxicity/ototoxicity. To overcome aforementioned limitations, the current study aimed to develop KS loaded PLGA-Vitamin-E-TPGS nanoparticles (KS-PLGA-TPGS NPs, to act as an efficient carrier for controlled delivery of KS. To achieve a substantial extension in blood circulation, a combined design, affixation of polyethylene glycol (PEG to KS-PLGA-TPGS NPs and adsorption of water-soluble chitosan (WSC (cationic deacetylated chitin to particle surface, was raised for surface modification of NPs. Surface modified NPs (KS-PEG-WSC NPs were prepared to provide controlled delivery and circulate in the bloodstream for an extended period of time, thus minimizing dosing frequency. In vivo pharmacokinetics and in vivo biodistribution following intramuscular administration were investigated. NPs surface charge was close to neutral +3.61 mV and significantly affected by the WSC coating. KS-PEG-WSC NPs presented striking prolongation in blood circulation, reduced protein binding, and long drew-out the blood circulation half-life with resultant reduced kidney sequestration vis-à-vis KS-PLGA-TPGS NPs. The studies, therefore, indicate the successful formulation development of KS-PEG-WSC NPs with reduced frequency of dosing of KS indicating low incidence of nephrotoxicity/ototoxicity.

  18. Comparison of Boric Acid and Combination Drug of Polymyxin, Neomycin and Hydrocortisone (polymyxin NH) in the Treatment of Acute Otitis Externa.

    Science.gov (United States)

    Amani, Soroush; Moeini, Mohammad

    2016-07-01

    Acute otitis externa is an inflammation of the external auditory canal known as "swimmer's ear". Direct costs including medical treatment, painkillers, antibiotics, steroids or both and indirect costs are also remarkable. The aim of this study was to compare the effect of boric acid and polymyxin, neomycin and hydrocortisone composition in the treatment of acute otitis externa. This randomized clinical trial was carried out on 80 patients aged more than 17-year-old who were referred to Kashani hospital clinic with a diagnosis of acute otitis externa by otolaryngologist. The patients were randomly allocated to two groups (A: Boric acid and B: polymyxin NH ear drops) and Painkiller was prescribed and administered orally for all patients and in the presence of fever, cellulitis around the ears and neck adenopathy, broad-spectrum systemic antibiotics were used besides topical treatment. Symptoms of patients who were evaluated by a physician includes pain, discharge from the ear, swelling of the ear canal, auricle swelling, tenderness, and ear itching. In addition, pain was evaluated in patients and was recorded by Macgill Pain Questionnaire, in the first, third, seventh and tenth days. Results showed that itching on third day (p=0.007) and swelling of the ear canal in the examination of the third day (p=0.006) and the seventh day (p=0.001) in the polymyxin NH group was more than those of boric acid group. Overall mean pain based on McGill questionnaire was 11.10±1.49 in boric acid group in the examination on the first day and was 4.05±0.22 in the examination on the tenth day and in the polymyxin NH group, it was 10.9±0.99 on the first day and 4.20±0.40 on the tenth day. In both groups, pain relief was the same and there was no significant difference between two groups (p=0.075). The findings of this study showed slight differences in the effectiveness of the boric acid drug and combination of polymyxin, neomycin and hydrocortisone in the treatment of patients with

  19. 75 FR 4692 - Ophthalmic and Topical Dosage Form New Animal Drugs; Miconazole, Polymixin B, and Prednisolone...

    Science.gov (United States)

    2010-01-29

    ... strains of yeast (Malassezia pachydermatis) and bacteria (Staphylococcus pseudintermedius). The NADA is... (Staphylococcus pseudintermedius). (3) Limitations. Federal law restricts this drug to use by or on the order of...

  20. Cre/loxP-mediated excision of a neomycin resistance expression unit from an integrated retroviral vector increases long terminal repeat-driven transcription in human hematopoietic cells.

    Science.gov (United States)

    Fernex, C; Dubreuil, P; Mannoni, P; Bagnis, C

    1997-01-01

    Recombinant retroviruses are currently the most attractive vehicles for gene transfer into hematopoietic cells. Retroviral vectors often contain an easily selectable marker gene in addition to the gene of interest. However, the presence and selection for expression of the selectable gene often result in a significant reduction of the expression of the gene of interest in the transduced cells. In order to circumvent this problem, we have developed a Cre/loxP recombination system for specific excision of the selectable expression unit from integrated retroviruses. A retroviral vector, containing both a neomycin resistance expression unit flanked by loxP sites and granulocyte-macrophage colony-stimulating factor cDNA, was used to transduce the human hematopoietic K-562 cell line. Four transduced cell clones were then superinfected with a retrovirus containing a Cre recombinase expression unit. Molecular analyses of 30 doubly transduced subclones showed a strict correlation between cre expression and loxP-flanked selectable cassette excision, thus implying that Cre recombinase activity is very efficient in a retroviral context. Moreover, the excision of the selectable cassette results in a significant increase of granulocyte-macrophage colony-stimulating factor transcription driven by the retroviral promoter. PMID:9311833

  1. The control of accuracy during protein synthesis in Escherichia coli and perturbations of this control by streptomycin, neomycin, or ribosomal mutations.

    Science.gov (United States)

    Brakier-Gingras, L; Phoenix, P

    1984-05-01

    This review surveys the different experimental approaches which describe the binding of tRNA to mRNA-programmed ribosomes and the control of tRNA selection. This selection is best described by the two-step model proposed by Hopfield and demonstrated by Thompson and his collaborators. The model involves a first control at the initial reversible binding of tRNA to the ribosome and a second control, the proofreading control, which promotes rejection of the incorrect tRNA from a high-energy intermediate during the transition from the initial to the final binding state. Streptomycin, neomycin, and ribosomal fidelity mutations appear to affect both control steps. Their effect can be related to the location of the mutated ribosomal proteins and to the conformational changes induced in the ribosome by the misreading agents. An alteration of the first control probably results from a distortion of the codon-anticodon interaction, while an alteration of the second control may be caused by a change in the association between ribosomal subunits.

  2. Thermodynamic insights into drug-surfactant interactions: Study of the interactions of naporxen, diclofenac sodium, neomycin, and lincomycin with hexadecytrimethylammonium bromide by using isothermal titration calorimetry.

    Science.gov (United States)

    Choudhary, Sinjan; Talele, Paurnima; Kishore, Nand

    2015-08-01

    The success of drug delivery depends on the efficiency of the route of administration, which in turn relies on properties of the drug and its transport vehicle. A quantitative knowledge of association of drugs with transport vehicles is lacking when the latter are in the category of self assembled structures. The work reported in this manuscript addresses the mechanism of partitioning of naproxen, diclofenac sodium, neomycin and lincomycin in the micelles of hexadecytrimethylammonium bromide and that is quantitatively based on the measurement of thermodynamic parameters of interactions by using isothermal titration calorimetry. The addressed mechanism of partitioning is based on the identification of the type of interactions of these drugs with the surfactant micelles and monomers, along with the effect of the former on the micellization properties of the surfactant. The conclusions are based on the interpretation of the values of partitioning constant, standard molar enthalpy change, standard molar entropy change and the stoichiometry of the interaction. The results of this study have implications for deriving guidelines for the target oriented synthesis of new drugs that are to be used for effective delivery via micellar media. Copyright © 2015 Elsevier B.V. All rights reserved.

  3. 新霉素阻断ELISA试剂盒的研制与应用%Development and Evaluation of Blocking ELISA Kit for Neomycin Detection

    Institute of Scientific and Technical Information of China (English)

    王爱萍; 李发弟; 胡骁飞; 曾巧英; 李金录; 郭艳丽; 赵启法; 陈玉梅; 范璐

    2011-01-01

    本研究旨在建立新霉素(Neomycin,NEO)免疫学检测试剂盒.以制备的抗NEO单克隆抗体(NEO mAb)为基础,建立阻断ELISA分析方法,组装新霉素阻断ELISA试剂盒,并对试剂盒的性能进行鉴定.结果表明,NEO试剂盒(NEO-kit)标准曲线呈典型的S型,符合4参数logit曲线拟合,相关系数R2=0.990 3;检测范围为1.0~64.0μg·L-1,半数抑制浓度(IC50)为2.59 μg·L-1,灵敏度为0.75 μg·L-1,检测限为1.0 μg·L1.试剂盒除了和NEO阳性样品反应呈阳性外,与庆大霉素、链霉素、土霉素、沙丁胺醇、环丙沙星、二氟沙星、磺胺嘧啶及磺胺甲噁唑等其他药物均无交叉反应性.奶样、饲料样及肉样的平均添加回收率为89.50%、89.58%和87.92%;平均批内和批间变异系数均小于15%,而且批间变异系数小于批内变异系数;基质对NEO-Kit的检测结果影响比较小;NEO-Kit在4℃可保存6个月以上.试剂盒和HPLC-MS-MS对牛奶中新霉素回收率没有显著差异(P≥0.05).试剂盒和HPLC-MS-MS对饲料样检测结果也无显著差异(P≥0.05).本研究成功研制出敏感、特异、准确的NEO检测试剂盒.%A blocking ELISA kit for detection of neomycin was developed based on the NEO mAb, and its properties were examined. The result showed that the calibration curve of NEO-kit was a typical sigmoid curve fitted to the four parameters logistic equation with the linear detection of 1.0 to 64.0 μg · L-1 and R2 was 0. 990 3. The kit has a sensitivity of 0. 75 μg · L-1 , the IC50 of 2. 59 μg · L-1and the detection limit of 1. 0 μg · L-1. The kit had no cross-reactivity with gentamicin, streptomycin, oxytetracycline, ciprofloxacin, difloxacin, salbutamol, sulfadiazine and sulfamethoxazole. The recoveries of NEO in milk, feed and meat were 89. 50%, 89.58% and 87. 92%, respectively. The inter-assay and intra-assay coefficient variation were all below 15%.Moreover, the inter-assay coefficient variation was lower than that of

  4. 荧光法检测鸭肉中新霉素残留的研究%A Study on Determination of Neomycin Residue in Duck by Fluorescence Method

    Institute of Scientific and Technical Information of China (English)

    徐将; 刘木华; 袁海超; 肖海斌; 赵进辉

    2013-01-01

    Health problems caused by overuse of antibiotics have become the focus of attention.In order to detect antibiotic residues rapidly;fluorescence method was used to detect the neomycin residues in duck.A chemical reaction of neomycin with ortho-phthalaldehyde,mercaptoethanol was involved in the emulsifier OP -10 environment and strong fluorescence was produced after excitation light irradiation.With neomycin as the object of the study,fluorescence intensity of its own and the reactant was studied.The results indicated that the best fluorescence intensity was measured after 90 min by adding 1.5 mL o-phthalaldehyde solution,1 mL emulsifier OP-10 solution,2.5 mL mercapto ethanol solution;Neomycin standard samples of concentration range in 1.00 ~ 8.00 μg/mL were measured to obtain the linear equation F =45.3C + 9.7,thecorrelation coefficient was 0.984 9 ; duck extracted solution with different concentrations of neomycin was determined,the correlation coefficient was 0.9995.The results showed that the fluorescence method was a convenient and rapid analytical method for detection of neomycin residue.%抗生素滥用所导致的健康问题已成为人们关注的焦点.为了快速检测抗生素残留,利用荧光法对鸭肉中新霉素的残留进行检测.在乳化剂OP-10的环境下,新霉素和邻苯二甲醛、巯基乙醇发生化学反应,经过激发光照射后可以产生强的荧光.以新霉素为研究对象,研究了其自身和反应物的荧光强度.研究结果表明:加入1.5mL邻苯二甲醛溶液,1mL乳化剂OP-10溶液,2.5mL巯基乙醇溶液,并且反应90 min后测得的荧光强度为最佳;对浓度范围在1.00~8.00 μg/mL的新霉素标准样品进行测定,得到的线性方程为,相关系数为0.984 9;对含不同浓度新霉素的鸭肉提取液进行测定,相关系数为0.9998.结果表明,荧光法是一种方便、快速的新霉素残留分析方法.

  5. 巨桉无性系Eg5的卡那霉素和头孢霉素敏感性研究%Study on kanamycin and cefotaxime sensitivity of Eucalyptus grandis clone Eg5

    Institute of Scientific and Technical Information of China (English)

    郭利军; 曾炳山; 刘英; 李湘阳; 裘珍飞

    2012-01-01

    By taking Eucalyptus grandis colne Eg5 leaves as explants, the effects of Kanamycin sulfate on the callus proliferation, bud differentiation and shoot rooting of leaf explains of Eucalyptus grandis clone Eg5 and the effect of Cefotaxime on explant's regeneration and the growth of Agrobacterium tumefaciens GV3101 were studied. The results showed that the concentration of kanamycin for restraining callus proliferation was 17.5 mg/L, that for fully suppressing bud differentiation was 60 mg/L and that for inhibiting shoots rooting was 400 mg/L. Cefotaxime at a dosage from 0 mg/L to 300 mg/L had minimal impact on the differentiation of leaf explants. With a soak of the explants in the solution of 500 mg/L Cefotaxime, the growth of agrobacterium can be fully inhibited at a Cefotaxime dosage of 100 mg/L in medium.%以巨桉无性系Eg5叶片为外植体材料,试验了硫酸卡那霉素(kanamycin sulfate,Km)对外植体愈伤组织诱导增殖、芽分化、芽生根的影响,并且试验了头孢霉素(Cefotaxime,Cef对外植体再生的影响及其对根癌农杆菌GV3101的抑制效果.结果表明:抑制愈伤组织增殖培养的卡那霉素浓度为17.5 mg/L,完全抑制芽分化的卡那霉素浓度为60 mg/L,完全抑制生根的卡那霉素浓度为400 mg/L; Cef在0~500mg/L范围内,0~300mg/L Cef对外植体的生长分化影响最小;共培养结束后的外植体经过500mgL的Cef处理后,共培养基中添加100 mg/L的Cef便可以完全抑制农杆菌GV3101的生长.

  6. Susceptibility Testing of the Combination of enrolfoxacin and kanamycin against Some Pathogenic Bacteria of Dairy Cow Mastitis%恩诺沙星与硫酸丁胺卡那霉素对奶牛乳房炎常见病原菌的联合药敏试验

    Institute of Scientific and Technical Information of China (English)

    王爽; 张鹏宇; 刘宇; 王岩; 李丹; 朱丹丹; 和翀翼; 史同瑞

    2014-01-01

    本试验对恩诺沙星和硫酸丁胺卡那霉素在4种奶牛乳房炎常见病原菌(无乳链球菌、化脓链球菌、金黄色葡萄球菌、致病性大肠埃希氏菌)联合药敏试验中的相互作用进行了研究。结果表明,恩诺沙星和硫酸丁胺卡那霉素对各菌的MIC为8~64μg/mL,二者联合后的FIC指数范围为0.5~1,二者联合用药时为协同作用或相加作用。%To study the interaction between enrolfoxacin and kanamycin against the bacteria of dairy cow mastitis, the susceptibility testing of drug combination to Streptococcus agalactiae, Streptococcus pyogenes, Staphylococcus aureus and E.coli in vitro was conducted. The minimal inhibitory concentration(MIC) of enrolfoxacin and kanamycin against the batertia was determined with the grade dilution method. The fractional inhibitory concentration(FIC) was measured with checkerboard microdilution method. The results showed MIC of enrolfoxacin and kanamycin against the batertia was 8~64μg/mL and the FIC of enrolfoxacin and kanamycin was 0.5~1. The study suggested that the combination of enrolfoxacin and kanamycin to Streptococcus agalactiae, Streptococcus pyogenes and E.coli had additive effect, and to Staphylococcus aureus demonstrated synergic action.

  7. The resistance of Salmonella spp. isolates from Alabio duck against several antibiotics

    OpenAIRE

    1998-01-01

    A total of 172 isolates of Salmonella spp. consisted of S. typhimurium (70), S. Radar (52), S. senftenberg (25), S. Virchow (14), and S. amsterdam (11) from Alabio duck hatcheries in the District of Hulu Sungai Utara, South Kalimantan, were examined their resistencies against seven kinds of antibiotics, consisted of chloramphenicol, neomycin, trimethoprim, streptomycin, tetracycline, ampicillin, and polymixin B, by using agar disc diffusion method. The results showed that 70 isolates of S. ty...

  8. Neomycin, Polymyxin, and Bacitracin Ophthalmic

    Science.gov (United States)

    ... hands thoroughly with soap and water. Use a mirror or have someone else apply the ointment. Avoid ... symptoms, call your doctor immediately: eye pain irritation, burning, itching, swelling, or redness of the eye or ...

  9. Development of lateral line system and its resistance to neomycin in transgenic zebrafish%转基因斑马鱼侧线系统的形成及其对新霉素的抗性

    Institute of Scientific and Technical Information of China (English)

    邵贝丽; 严继舟

    2014-01-01

    In this experiment gene -targeting GFP transgenic zebrafish 189 b was used to observe the whole development process of lateral line system and the responses to neomycin treatment.The results showed that the occurrence and distribu-tion of GFP-labeling cells were highly consistent with the spatiotemporal development of the neuromasts along the lateral line, especially in migration route and final residence.As the larvae grew , the diffused migratory GFP -cells focused as small and bright GFP-dots.Exposure of 24 h and 6 d of GFP-positive embryos in 150 μmol/L of amino sugar glucoside class antibiotic ( neomycin ) did not change the GFP -pattern and distribution.These results suggested that the GFP -labe-ling cells might be neuromast progenitors and resistant to neomycin.%利用转基因斑马鱼(Danio rerio)189b动态观察侧线系统形成及其对新霉素处理的敏感性。试验结果显示,转基因斑马鱼荧光细胞出现、迁移和定居与侧线神经丘的形成走向和发育时间基本吻合。随着发育时间的推移,荧光点的形态也发生了较大变化,由最初的比较模糊的聚集在一起的荧光团,逐渐变成颗颗分明的荧光点,大小也由大变小。将受精后24 h和6 d带有荧光的细胞暴露于150μmol/L的氨基糖甙类抗生素(新霉素)中,并不影响荧光细胞的数量和分布。结果说明荧光标记细胞对新霉素处理不敏感,不是毛细胞,可能是神经丘形成的先祖细胞。

  10. The resistance of Salmonella spp. isolates from Alabio duck against several antibiotics

    Directory of Open Access Journals (Sweden)

    Istiana

    1998-06-01

    Full Text Available A total of 172 isolates of Salmonella spp. consisted of S. typhimurium (70, S. Radar (52, S. senftenberg (25, S. Virchow (14, and S. amsterdam (11 from Alabio duck hatcheries in the District of Hulu Sungai Utara, South Kalimantan, were examined their resistencies against seven kinds of antibiotics, consisted of chloramphenicol, neomycin, trimethoprim, streptomycin, tetracycline, ampicillin, and polymixin B, by using agar disc diffusion method. The results showed that 70 isolates of S. typhimurium were resistant against six kinds of antibiotics with various percentages, that is chlorampenicol 5 .7%, neomycin 12 .8%, trimethoprim 7.1 %, streptomycin 8 .6%, tetracycline 11 .4%, and the highest against ampicillin 30 %. Ofthe 52 S. hadar isolates showed that all ofthem were resistent against 6 kinds of antibiotics, that is chloramphenicol 1 .9%, neomycin 7.7%, trimethoprim 5.8%, streptomycin 48 .0%, tetracycline 48.0%, and polymixin B 3.8%. Futhermore S. senftenberg (25 isolates were resistent against three kinds of antibiotics, that is neomycin 12%, streptomycin 20%, and tetracyclines 16%. Meanwhile S. virchow (14 isolates, were resistant against two kinds of antibiotics namely streptomycin 7.1% and tetracylines 14.3%. Whereas S. amsterdam (11 isolates were resistant against four kinds of antibiotics, that is neomycin 45 .5%, streptomycin 18 .2%, tetracycline 18 .2%, and ampicillin 9.10%.

  11. calpain1在卡那霉素致毒豚鼠耳蜗中的表达%Expression of calpain 1 in kanamycin-poisoned guinea pig cochlea

    Institute of Scientific and Technical Information of China (English)

    马德菊

    2011-01-01

    Aim To investigate the expression of calpain 1 in kanamycin( KM )-poisoned guinea pig cochlea. Methods Guinea pigs were randomly assigned to four groups : control group, KM 3 days group , 7 days group and 14 days group. Auditory brainstem response ( ABR )measurement was used to test the hearing of guinea pig. The expression of calpain I was detected by the SABC method of immunohistochemistry combined with imaging analysis technique. Auditory brainstem response( ABR )measurement was used to test the hearing of guinea pig. Results Immunoreactivity for calpain 1 was mainly found in hair cells , spiral ganglions, spiral ligament and stria vascularis. The results of imaging analysis indicated that immuncorectivity for calpain 1 gradually lowered with the days up. Conclusion Calpain 1 is found in Cochlea of control group and KM group. The results of imaging analysis indicate that immuncorectivity for calpain 1 gradually lower with the days up.%目的 探讨钙蛋白酶(calpain 1)在卡那霉素(kanamycin,KM)致毒豚鼠耳蜗中的定位与表达.方法 将40只豚鼠随机均分成4组:对照组、KM 3 d组、KM 7 d组和KM 14 d组,KM各组每天肌肉注射硫酸卡那霉素注射液200 mg·kg-1.应用电生理指标听性脑干反应(auditory brainstem response,ABR)观察用药前后豚鼠听力的变化;采用免疫组织化学SABC法结合显微图像分析技术,观察豚鼠耳蜗组织中calpain 1的表达.结果 calpain 1阳性免疫反应主要见于耳蜗毛细胞、螺旋神经节、螺旋韧带和血管纹,并随给药天数的增加,calpain 1在耳蜗上述部位的表达逐渐减弱.结论 calpain 1在正常豚鼠和卡那霉素致毒豚鼠耳蜗中都有表达,且随卡那霉素给药天数的增加表达逐渐减弱.

  12. Expression of calpains in the cochlea in kanamycin-poisoned guinea pig%钙蛋白酶在卡那霉素致毒豚鼠耳蜗中的表达

    Institute of Scientific and Technical Information of China (English)

    马德菊; 王爱梅

    2011-01-01

    目的 探讨钙蛋白酶(calpain)在卡那霉素(kanamycin,KM)致耳中毒豚鼠耳蜗的表达.方法 将豚鼠随机分成对照组、KM 3 d组、KM 7 d组和KM 14 d组,应用免疫组织化学SABC(streptavidin-biotin peroxidae complex,链霉亲合素-生物素过氧化物酶复合物)法和显微图像分析技术检测耳蜗中钙蛋白酶的表达,用药前后给予短纯音刺激检测听性脑干反应阈值,观察豚鼠听力的变化.结果 对照组calpain 1阳性免疫反应主要见于耳蜗毛细胞、螺旋神经节、血管纹和螺旋韧带,以螺旋神经节的染色较深,而其它部位均呈阴性.肌肉注射KM后,calpain 1在耳蜗中的阳性反应部位与对照组大致相同,显微图像分析结果表明,随着给药天数的增加,calpain 1在耳蜗上述部位的阳性反应逐渐减弱.Calpain 2在各组豚鼠耳蜗中的表达部位与calpain 1的相同,显微图像分析结果提示,随着给药天数的增加,calpain 2在耳蜗上述部位的阳性反应逐渐增强.结论 正常豚鼠耳蜗中有calpain1和calpain 2的表达.注射KM后,随着给药天数的增加,calpain 1在耳蜗的表达逐渐减弱,而calpain 2的表达则逐渐增强,提示calpain 2可能参与了卡那霉素致耳中毒的过程.%Objective To investigate expression of calpains in the cochlea in kanamycin (KM) -poisoned guinea pigs. Methods Guinea pigs were randomly assigned to serve as controls or to receive KM for 3, 7 or 14 days. SABC im-munohistochemistry and imaging analysis were used to examine expression of calpain 1 and calpain 2. Auditory brainstem responses (ABRs) were used to determine auditory thresholds and their changes. Results Immunoreactivity for clapin 1 was mainly seen in the hair cell, spiral ganglion, stria vascularis and spiral ligament in the control group, with greater staining in the spiral ganglion. The location of calpain 1 expression in the cochlea was similar in the control and KM treatment groups. Imaging analysis indicated that

  13. 新霉素快速半定量ELISA试剂盒的研制与应用%Development and Apply of Semi-quantitative Quick ELISA Analysis for Neomycin

    Institute of Scientific and Technical Information of China (English)

    邢广旭; 王方雨; 胡骁飞; 邓瑞广; 张改平

    2013-01-01

    Basing on the monoclonal antibody of neomycin,horse radish peroxidase (HPR) was coupled.Enzyme Linked Immunosorbent Assay (ELISA) kit for detection NEO was established by SandwichBlocking mode.The results showed that there was no cross reaction with other analogue of NEO,and the limit of detection (LOD) in milk,feed and muscle were 1 microgramme per litre,and expiration date was more than 6 month.The results were no significant difference between HPLC-MS-MS and the kit.This kit helps to strengthen the neomycin drug illegal abuse,and support a powerful technical method in order to improve our country animal food safety and protect the health of the people.%本研究利用已经筛选的抗新霉素单克隆抗体,直接与辣根过氧化物酶相偶联,确定其相应的工作浓度,建立了快速半定量ELISA试剂盒.结果表明,本研究研制的新霉素(NEO)阻断ELISA试剂盒可在30min内进行半定量检测,其特异性较强,与其它氨基糖苷类抗生素的交叉反应均比较低;试剂盒的在奶样、饲料和肌肉样品的检测限均为1 μg·L-1;NEO-Kit在4℃至少可以保存6个月以上;试剂盒与液相色谱-二级质谱联用方法检测新鲜奶样中NEO含量的结果比较之间无显著差异;对相同浓度的NEO样品回收率无显著差异.NEO快速半定量ELISA试剂盒的成功研制有助于进行现场样品检测,为检测NEO药物的滥用提供了新的快捷检测手段.

  14. Transformation of Neomycin Resistance Gene (neoR) into Silkworm (Bombyx mori.L.)%新霉素抗性基因在家蚕中的插入和表达

    Institute of Scientific and Technical Information of China (English)

    陈秀; 赵昀; 张峰; 彭卫平; 冯晓黎; 黄君霆; 陆长德

    1999-01-01

    构建含新霉素抗性基因(neomycin resistance gene, neoR)的重组质粒pFN, 经HindIII酶切后, 用基因枪将DNA片段导入家蚕早期受精卵中(G0代). 孵化的G1、G2代蚁蚕均经含新霉素的人工饲料添食24 h后, 筛选出新霉素抗性的个体(能正常生长发育的)改为桑叶饲养. 于G2代的5龄第二天从后部丝腺抽提总DNA, 再以neoR的cDNA为探针进行Southern杂交检测. 结果表明neoR基因已转入家蚕DNA中, 获得了含neoR的转基因蚕.

  15. Agrobacterium tumefaciens-mediated transformation of Vigna mungo (L.) Hepper.

    Science.gov (United States)

    Karthikeyan, A S; Sarma, K S; Veluthambi, K

    1996-01-01

    Transformed Vigna mungo (blackgram) calli were obtained by cocultivating segments of primary leaves with Agrobacterium tumefaciens vir helper strains harbouring the binary vector pGA472 having kanamycin resistance gene as plant transformation marker. Transformed calli were selected on Murashige and Skoog medium supplemented with 50 mg/l kanamycin and 500 mg/l carbenicillin. Transformed calli were found to be resistant to kanamycin up to 900 mg/l concentration. Expression of kanamycin resistance gene in transformed calli was demonstrated by neomycin phosphotransferase assay. Stable integration of transferred DNA into V. mungo genome was confirmed by Southern blot analysis.

  16. 麻腮风联合减毒活疫苗中新霉素残留量微生物学检测方法的建立%Development of a microbiological method for detection of residual neomycin content in live attenuated measles, mumps and rubella combined vaccine

    Institute of Scientific and Technical Information of China (English)

    常艳; 杨美琴; 李景云; 胡昌勤

    2013-01-01

    目的 建立麻腮风联合减毒活疫苗(Measles,mumps and rubella vaccine,MMR)中新霉素残留量的微生物学检测方法,并进行验证.方法 采用管碟法测定新霉素含量,并以新霉素浓度的对数和抑菌圈半径的平方值绘制标准直线.对建立的方法进行同质性、最低检出限、加样回收率及精密度验证.结果 建立的方法在新霉素浓度为0.24~4.18U/ml的范围内线性关系良好,R2=0.997 8;新霉素标准品与供试品的剂量反应直线的回归方程的斜率差异无统计学意义(P>0.05),即标准品与供试品间可满足同质性的要求;该方法的最低检出限为0.05 U/ml;该方法检测新霉素浓度约为0.5和1.0 U/ml的混合溶液,回收率分别为99.22%和99.86%;日内RSD为1.09%,日间RSD为1.42%.结论 建立了一种适用于定量测定MMR中新霉素残留量的微生物学检测方法,该方法操作简便,结果可靠,可用于MMR的常规质量控制.%Objective To develop and verify a microbiological method for detection of residual neomycin content in live attenuated measles, mumps and rubella combined vaccine (MMR). Methods The neomycin content was determined by cylinder plate method, based on which a standard curve was plotted with the log of neomycin concentration against the square of radius of bacteriostatic ring. The developed method was verified for homogeneity, limit of detection (LOD), recovery rate and precision. Results The developed method showed good linearity within a neomycin concentration range of 0. 24 ~ 4. 18 U/ ml (R2 - 0. 997 8). No significant difference was observed between the slopes of dose-response curves of standard neomycin and test samples (P> 0. 05), which met the requirements for homogeneity. The LOD of the developed method was 0. 05 U / ml. By the developed method, the recovery rates of mixed samples at neomycin concentrations of 0. 5 and 1. 0 U/ml were 99. 22% and 99. 86%, while the intra- and inter-RSDs were 1. 09

  17. Math1 gene therapy for kanamycin and furosemide-induced deaf guinea pigs%卡那霉素联合速尿致聋豚鼠的Math1基因治疗

    Institute of Scientific and Technical Information of China (English)

    张贤芬; 杨仕明; 韩东一; 郭维维; 孙建和; 高举; 孙东绣; 孙爱玲; 李臻

    2013-01-01

    Objective To observe the morphology and function changes of cochlear hair cells before and after math1 gene injection into the cochlea of deaf guinea pigs which were induced by kanamycin and furosemide.To explore the feasibility of Math1 gene for medicine-induced deafness therapy.Methods Kanamycin(500 mg/kg) and furosemide(50 mg/kg) were given to the healthy adult guinea pigs intramuscularly and intravenously to establish the deafness model.The guinea pigs whose auditory brainstem response (ABR) threshold > 95 dB SPL were randomly divided into five groups.Blank control group (without any treatment,n =3),operation control group (right ear scala tympani operation,n =3),artificial perilymph group (right ear scala tympani injection artificial perilymph,n =3),virus vector group [right ear scala tympani injection adenovirus which carrying enhanced green fluorescent protein(EGFP) gene (Ad.EGFP),n =4],Math1 gene therapy group [right ear scala tympani injection adenovirus which carrying Math1 and EGFP gene(Ad.Math1-EGFP),n =6].Each animal received ABR test before and after injection.The cochlear tissue was observed by scanning electronic microscopy.Results The ABR thresholds of tone burst(4,8,16,20 kHz) were not statistically significant in different groups (P > 0.05).The number of hair cells increased in some of severe deaf guinea pigs after the injection of Ad.Math1-EGFP gene.However,there was no obvious difference with morphology and numbers of cochlea hair cells in other groups.Conclusions The injection of Math1 gene to cochlea can regenerate or repair the hair cells of medicine-induced deaf guinea pigs,but there was no improvement on the hearing loss.%目的 观察卡那霉素和速尿联合致聋豚鼠耳蜗鼓阶导入Math1基因后的形态学及功能改变,探讨Mathl基因治疗药物中毒性耳聋的可行性.方法 健康成年豚鼠经硫酸卡那霉素(500 mg/kg)和速尿(50 mg/kg)联合致聋,将听性脑干反应(ABR)反应阈>95 dB SPL的豚鼠按

  18. Multiple drug resistance of Aeromonas hydrophila isolates from Chicken samples collected from Mhow and Indore city of Madhyapradesh

    Directory of Open Access Journals (Sweden)

    Kaskhedikar

    2009-02-01

    Full Text Available Fourteen antibacterial agents belonging to 9 different groups of antibiotics viz. aminoglycosides, cephalosporins, nitrofurantoin, fluroquinolones, chloramphenicol, sulphonamides, tetracyclines, penicillin and polymixin were used for in vitro sensitivity testing of Aeromonas hydrophila isolated from fifteen samples of chicken collected from retail shops in Mhow city. The sensitivity (100% was attributed to ciprofloxacin, cefuroxime, ceftriaxone, cephotaxime, chloramphenicol, gentamycin, kanamycin, nitrofurantoin, nalidixic acid and ofloxacin followed by oxytetracycline (50%. All the isolates were resistant to ampicillin and colistin antibiotics. That means, none of the isolates were found to be sensitive for penicillin and polymixin group of antibiotics. Multiple drug resistance was also observed in all A. hydrophila isolates. Out of total isolates, 100% were resistant to two antimicrobial drugs and 50% to three drugs. [Vet. World 2009; 2(1.000: 31-32

  19. Gene trap mutagenesis of hnRNP A2/B1: a cryptic 3' splice site in the neomycin resistance gene allows continued expression of the disrupted cellular gene

    Directory of Open Access Journals (Sweden)

    DeGregori James V

    2003-01-01

    Full Text Available Abstract Background Tagged sequence mutagenesis is a process for constructing libraries of sequenced insertion mutations in embryonic stem cells that can be transmitted into the mouse germline. To better predict the functional consequences of gene entrapment on cellular gene expression, the present study characterized the effects of a U3Neo gene trap retrovirus inserted into an intron of the hnRNP A2/B1 gene. The mutation was selected for analysis because it occurred in a highly expressed gene and yet did not produce obvious phenotypes following germline transmission. Results Sequences flanking the integrated gene trap vector in 1B4 cells were used to isolate a full-length cDNA whose predicted amino acid sequence is identical to the human A2 protein at all but one of 341 amino acid residues. hnRNP A2/B1 transcripts extending into the provirus utilize a cryptic 3' splice site located 28 nucleotides downstream of the neomycin phosphotransferase start codon. The inserted Neo sequence and proviral poly(A site function as an 3' terminal exon that is utilized to produce hnRNP A2/B1-Neo fusion transcripts, or skipped to produce wild-type hnRNP A2/B1 transcripts. This results in only a modest disruption of hnRNPA2/B1 gene expression. Conclusions Expression of the occupied hnRNP A2/B1 gene and utilization of the viral poly(A site are consistent with an exon definition model of pre-mRNA splicing. These results reveal a mechanism by which U3 gene trap vectors can be expressed without disrupting cellular gene expression, thus suggesting ways to improve these vectors for gene trap mutagenesis.

  20. 新唑漱口液微生物限度检查方法的验证%Validation of Microbial Limit Tests of Neomycin and Metronidazole Garga-rism

    Institute of Scientific and Technical Information of China (English)

    余晓霞; 邱凯锋; 刘春霞

    2014-01-01

    OBJECTIVE To establish a method to test microbial limit in Neomycin and Metronidazole garga-rism.METHODS Microbial limits test in Chinese Pharmacopoeia 2010 ( section 2 ) appendix verified methodology was used.RESULTS The durg had no inhibitory action to white beads bacterium and aspergillus ,and Membrane filtration method can eliminate the drug on staphylococcus aureus ,e.coli,bacillus subtilis inhibitory effect.The recov-ery can reach up to 75%.CONCLUSION Through the technological validation ,the result is consistent with the re-quirement of ChP.( edition 2010 ) in force.%目的:建立新唑漱口液微生物限度检查方法。方法采用2010年版《中国药典》(二部)附录“微生物限度检查法”项下相关内容进行方法学验证。结果该漱口液对白色念珠菌和黑曲霉菌无抑制作用;薄膜过滤法可消除该药物对金黄色葡萄球菌、大肠埃希菌、枯草芽孢杆菌的抑制作用,回收率达75%以上。结论经方法学验证,结果符合现行《中国药典》2010年版的要求。

  1. STUDIES ON THE DETERMINATION OF NEOMYCIN RESIDUES IN CHICKEN TISSUES BY MICROBIOLOGICAL METHOD%鸡组织中新霉素药物残留的微生物学检测方法的研究

    Institute of Scientific and Technical Information of China (English)

    沈川; 肖希龙

    2000-01-01

    新霉素(Neomycin)是由链霉菌属弗氏链霉菌(Streptomyces fradiae)经发酵培养产生的一种氨基糖甙类抗生素,目前,它作为一种常用兽药和具有发展前景的饲料添加药物在畜牧业中广为使用.为了控制新霉素在动物性食品中的残留,保障食品卫生和公共健康,本研究首次在国内对鸡体内新霉素残留的微生物学检测方法进行了研究.本研究选择2倍于样本重量的0.1 mol/L磷酸盐缓冲液(pH8.0)为组织提取液,经加热、离心除去蛋白质,用测试菌为表皮葡萄球菌(Staphylococcus epidermidis)的微生物学杯碟法测定提取液中新霉素的含量.本方法的灵敏度为0.15~0.2 μg/mL,检测限为0.5~0.75μg·g-1组织,回收率为85.3%~93.2%,回收率比FDA的传统方法有显著提高.

  2. Effect of neomycin on cell proliferation and expression of PDGF,VEGF,angiogenin in glioma cells%新霉素对脑胶质瘤细胞增殖及PDGF、VEGF和血管生成素表达的影响

    Institute of Scientific and Technical Information of China (English)

    赵佳; 郝利铭; 姜文华; 孔德霞; 李洪成; 周莉

    2014-01-01

    目的:探讨新霉素对脑胶质瘤细胞增殖及 PDGF、VEGF和血管生成素表达的影响。方法采用人脑胶质瘤细胞 U251,DMEM培养基添加10%胎牛血清培养。MTT细胞活性实验检测细胞增殖,Real Time PCR检测 mR-NA表达情况,酶联免疫吸附实验检测蛋白表达。结果 MTT结果表明,新霉素对 U251细胞增殖存在抑制作用,并以剂量依赖的方式进行,并发现新霉素的抑制作用随时间增强。Real Time PCR结果显示,新霉素作用后 U251细胞中PDGF、VEGF和血管生成素 mRNA 的表达都受到不同程度的抑制,分别降低了26.75%、38.23%和43.87%。ELISA分析表明新霉素能够降低PDGF、VEGF和血管生成素蛋白水平的表达。结论新霉素能够抑制脑胶质瘤细胞 U251中PDGF、VEGF和血管生成素的表达并抑制脑胶质瘤细胞增殖。%Objective To observe the effect of neomycin on cell proliferation and expression of PDGF,VEGF,angio-genin in U251 glioma cells.Methods The cell proliferation was analyzed using MTT.Real Time PCR and ELISA were applied to investigate the expression of PDGF,VEGF,angiogenin.Results The MTT results showed that neomycin positively inhibited the cell proliferation of U251 cells and the inhibition was enhanced by dose-dependent and time-de-pendent.Real Time PCR and ELISA results showed that neomycin inhibited the expression of PDGF,VEGF,angiogenin both on mRNA level and protein level.Conclusion Neomycin positively inhibited the cell proliferation and expression of PDGF,VEGF,angiogenin in U251 glioma cells.

  3. Distribution of putative virulence genes and antimicrobial drug resistance in Vibrio harveyi

    Digital Repository Service at National Institute of Oceanography (India)

    Parvathi, A.; Mendez, D.; Anto, C.

    . monodon hatchery and grow out systems, where they can infect post larvae causing large scale mortalities [3, 4]. Infection with biofilm forming V. harveyi resistant to multiple antibiotics has been previously reported from India [5]. The pathogenic... resistant to neomycin. Since kanamycin, streptomycin and neomycin belong to aminoglycoside group of antibiotics, the resistance to multiple antibiotics of the same class is expected. Similarly, resistance to macrolids erythromycin and azithromycin, β...

  4. Determination of triamcinolone acetonide acetate in triamcinolone acetonide and neomycin paste by HPLC%HPLC法测定曲安奈德新霉素贴膏中醋酸曲安奈德的含量

    Institute of Scientific and Technical Information of China (English)

    胡守莲; 邱栋樑

    2015-01-01

    目的:建立曲安奈德新霉素贴膏中醋酸曲安奈德含量的HPLC测定方法。方法采用反相高效液相色谱法,使用DIONEX Acclaim 120 C18(250mm×4.6mm,5μm)色谱柱,以甲醇-水(60∶40)作为流动相,检测波长240nm,流速1.0mL/min,柱温为30℃。结果醋酸曲安奈德在0.05~0.50μg范围内线性较好(r=0.9999),平均回收率为98.09%(RSD为0.73%)。结论该方法快速、简便、可靠、准确度高、重复性好,处方中其他成分对测定亦无干扰,分离效果好,可用于曲安奈德新霉素贴膏中醋酸曲安奈德含量测定。%ObjectiveTo establish a method for content determination of triamcinolone acetonide acetate in triamcinolone acetonide and neomycin paste by HPLC.Methods The experimental condition of the RP-HPLC method was as follows:DIONEX Acclaim 120 C18(250mm×4.6mm,5μm),with linear gradient elution using methanol and water (60∶40),The detected wavelength was 240 nm,the flow rate was 1.0 ml/min,The column temperature was at 30℃. ResultsThe fluocinolone acetonide curve was linear in the range of 0.05~0.50μg(r=0.9999).The average recovery rate was 98.09% withRSD of 0.73%.ConclusionThe method is simple,reliable,accurate and It could be used in the determination of triamcinolone acetonide acetate in triamcinolone acetonide and neomyc in paste.

  5. Determination of Trace Neomycin Sulfate by Photoluminescence and Lanthanide Sensitized Fluorescence Quenching on Europium-norfloxacin-SDS%利用铕(Ⅲ)-诺氟沙星-十二烷基硫酸钠的光致发光及稀土敏化荧光法测定硫酸新霉素的浓度

    Institute of Scientific and Technical Information of China (English)

    刘保生; 潇娜; 种宝红; 曹世娜; 吕运开

    2012-01-01

    In the ammonia-ammonium chloride buffer solution of pH =7.4, europiumion ( Eu3 + ) and norfloxacin(NFLX) react and combine as steady complex under room temperature. The complex shows its sensitized fluorescence of rare earth. The maximum excitation wavelength and maximum emission wavelength are 340 nm and 612 nm, respectively. The fluorescence intensity has a further increase in sodium dodecyl sulfate (SDS). When neomycin sulfate solution is added, the fluorescence intensity of the system of Eu3 + -NFLX-SDS diminishes, while the locations of the excitation and emission wavelengths stay the same. According to this result, a new way to determine the concentration of neomycin sulfate is established. The concentration range that neomycin sulfate can detect is 0.909~22.7 mg/L. The detection limit is up to 0.4 mg/L. This measurement has been applied to determine neomycin sulfate in tablet and drops. The relative standard deviation is 1.4% ~ 3.5% for 6 parallel determination of neomycin sulfate samples and the recovery is 90. 5% ~ 102.3%.%室温下,在pH =7.4的氨-氯化铵缓冲溶液中,铕(Ⅲ)与诺氟沙星反应形成稳定络合物,产生稀土敏化荧光,其最佳激发、发射波长分别为λex=340 nm、λem=612 nm,表面活性剂十二烷基硫酸钠(SDS)的加入使其荧光强度进一步增加.在该反应体系中加入适量硫酸新霉素溶液,铕(Ⅲ)与诺氟沙星络合物的激发、发射峰位置不变,但荧光强度降低,据此建立了灵敏的测定硫酸新霉素的荧光分析方法.硫酸新霉素的浓度在0.909~ 22.72 mg/L范围内符合线性关系.该方法的检出限为0.40 mg/L,可用于片剂及滴眼液中硫酸新霉素含量的测定,快速、简便、结果满意.6次平行测定的回收率为90.5% ~ 102.3%,相对标准偏差为1.4% ~3.5%.

  6. Clinical Observation of Prurigo Nodularis Treated with Autohemotherapy Combined with Desloratadine and Triamcinolone-Acetonide-Neomycin Paste%自血疗法配合地氯雷他定、肤疾宁治疗结节性痒疹临床观察

    Institute of Scientific and Technical Information of China (English)

    郑慕雄; 李瑾娴; 许敏鸿

    2011-01-01

    目的 观察自血疗法配合口服地氯雷他定、肤疾宁治疗结节性痒疹的临床疗效及复发率.方法 将80例患者随机分为两组,治疗组40例采用自血疗法配合口服地氯雷他定片、外用肤疾宁贴膏治疗;对照组40例采用口服地氯雷他定片、外用肤疾宁贴膏治疗;疗程均为30 d.结果 治疗组痊愈15例,有效率82.05%;对照组痊愈9例,有效率58.97%,两组有效率比较,差异有统计学意义(P<0.05);治疗结束30 d后随访有效病例,治疗组复发8例,总复发率25.00%;对照组复发13例,总复发率56.52%,两组总复发率比较,差异有统计学意义(P<0.05).结论 采用自血疗法配合地氯雷他定、肤疾宁治疗结节性痒疹,能降低复发率,临床疗效明显优于单纯采用口服地氯雷他定片、外用肤疾宁贴膏,值得临床推广运用.%Objective To observe the clinical efficacy and recurrence rate of prurigo nodularis treated with autohemotherapy combined with oral administration of desloratadine as well as triamcinolone - acetonide - neomycin paste.Methods 80 cases were randomized into two groups.In a treatment( 40 cases ), autohemotherapy was adopted together with oral administration of desloratadine and topical application of triamcinolone - acetonide - neomycin paste.In a control group( 40 cases ), oral administration of desloratadine and topical application of triamcinolone - acetonide - neomycin paste were applied.The duration of treatment was 30 days in either group.Results 15 cases were cured in treatment group and the effective rate was 82.05%.9 cases were cured in control group and the effective rate was 58.97%.The difference was significant statistically in comparison of the effective rate between two groups( P < 0.05 ).The follow - up was conducted for the effective cases 30 days later after treatment.8 cases were recurred in treatment group and the total recurrence rate was 25.00%.13 cases were recurred in control group and the

  7. HPLC-ELSD法同时测定新氢化可的松乳膏中硫酸新霉素和氢化可的松的含量%Determination of neomycin sulfate and hydrocortisone in hydrocortisone acetate ointment by HPLC-ELSD

    Institute of Scientific and Technical Information of China (English)

    徐兴亚; 李倚云

    2011-01-01

    目的 建立HPLC-蒸发光散射检测器(ELSD)法,同时测定新氢化可的松乳膏中硫酸新霉素和氢化可的松的含量.方法 色谱条件:C18色谱柱(150 mm×4.6 mm,5 μm);流动相:乙腈-0.1 mol/L三氟乙酸溶液(梯度洗脱);漂移管温度:110 ℃;载气流速:2.6 L/min.用硫酸新霉素和氢化可的松对照品分别测定新氢化可的松乳膏中硫酸新霉素和氢化可的松的含量.结果 硫酸新霉素进样量在0.7~4.3 μg范围内(r=0.999 7),氢化可的松进样量在1.0~2.9 μg范围内(r=0.999 9).结论 该方法快速、简便、重现性好,灵敏度高.%Objective To determine the content of neomycin sulfate and hydrocortisone in hydrocortisone acetate ointment by HPLC-ELSD.Methods C 18 ( 150 mm × 4.6 mm,5 μm)column was used, mobile phase:acetonitrile0.1 mol/L trifluoroacetic acid(gradient elution) ,the temperature of drift tube was set at 110 ℃ ,and flow rate of gas was 2.6 L/min.The contents of neomycin sulfate and hydrocortisone in hydrocortisone acetate ointment were determined.Results The calibration curve of neomycin was linear in the range of 0.7 ~ 4.3 μg ( r = 0.999 7 ).The calibration curve of hydrocortisone was linear in the range of 1.0 ~ 2.9 μg ( r = 0.999 9 ).Conclusion The method is rapid,sensitive, accurate and convenient.

  8. Estudo comparativo de duas técnicas farmacopéicas de avaliação da atividade antimicrobiana dos fármacos: nistatina, eritromicina, neomicina e gentamicina Comparison of two pharmacopeical thecnics to the evaluation of the antimicrobial activity of the drugs: nystatin, erythromycin, neomycin and gentamicin

    Directory of Open Access Journals (Sweden)

    Tatiane Margato Vital

    2004-06-01

    Full Text Available Através das técnicas de difusão em ágar com discos de papel e cilindros de aço inoxidável, analisou-se quantitativamente a atividade antimicrobiana de 123 medicamentos contendo os fármacos: nistatina (43 amostras de creme vaginal, eritromicina (14 amostras de comprimidos e 9 amostras de suspensão oral, gentamicina (33 amostras de líquido injetável e neomicina (24 amostras de creme de uso tópico, mediante as seguintes cepas de microrganismos: Saccharomyces cerevisiae ATCC 2601 para a nistatina; Staphylococcus epidermidis 12228 para a gentamicina e neomicina e Micrococcus luteus ATCC 9341 para a eritromicina. O preparo das cepas, soluções de padrão e amostra e meios de cultura nos dois métodos seguiram as recomendações da Farmacopéia Brasileira IV (1988 e Farmacopéia Americana 24 (2000. Os resultados foram analisados estatisticamente pelos testes de Fisher (p=0,05 e Student (p=0,05. Dentre as 43 amostras de nistatina creme vaginal analisadas, verificaram-se pelo teste de Student , diferenças pouco significativas (para p=0,05 de atividade antimicrobiana entre os métodos dos discos e cilindros; porém no teste de Fisher não houve diferenças. Nas amostras de eritromicina, gentamicina e neomicina não se obtiveram diferenças significativas entre os testes estatísticos de Fisher e Student. Conclui-se que, para a eritromicina, gentamicina, neomicina e nistatina, a análise de atividade antimicrobiana pela técnica dos discos de papel pode substituir a técnica dos cilindros de aço inoxidável.Through the diffusion techniques in culture media with paper disc and stainless steel cylinders the antimicrobial activity in 123 pharmaceutical preparations was evaluated: nystatin (43 samples of vaginal cream, erythromycin (14 samples of tablets and 9 of oral suspension, gentamicin (33 samples of injectable liquid and neomycin (24 samples of cream of topic use, against the microorganisms: Saccharomyces cerevisiae ATCC 2601 to nystatin

  9. Topical antibiotics in the management of corneal ulcer

    Directory of Open Access Journals (Sweden)

    Reddy P

    1988-01-01

    Full Text Available A total of 82 patients suffering from corneal ulcer were treated with framycetin 0.5%, gentamicin 3 mg./ml, chloramphenicol 0.4% and a neomycin combination containing Polymixin B sulphate 5000 u, neomycin sulphate 1700 u and gramicidin 0.025 mg/mL in a Randomised comparative study. The commonest organism isolated was Staphylococcus followed by Pneumococcus, Streptococcus and Pseudomonas. The in vitro sensitivity of these isolates to framycetin was higher than that to others Framycetin produced both earlier and a greater degree of improvement in mean score of signs and symptoms than the other antibiotics. It can thus be concluded that framycetin has a better profile of antibacterial activity and clinical efficacy than some other commonly used topical antibiotics in the treatment of corneal ulcer.

  10. [Transmissivity of antibiotic resistance factors in intergeneric crossing of salmonellae and Escherichia Crimea].

    Science.gov (United States)

    Kharchenko, G I; Zadorina, T M; Belova, N N

    1981-11-01

    The results of the crosses between Salmonella and Escherichia 0151C are presented. Transmissive determinants resistant to ampicillin, dentamicin, kanamycin, neomycin, monomycin, streptomycin, morphocyclin, tetracycline and chloramphenicol were detected. It was shown that almost 50 per cent of Escherichia 0151K except the transmissive R factors were able to transferring the colicinogenic determinants.

  11. Possible Application of Biotechnology to the Development of Biological Agents by Potential Enemies

    Science.gov (United States)

    1987-06-01

    neomycin , and kanamycin. Unlike the enterobacteria, the Staphylococci can not conjugate (sexual exchange) so that plasmids must be transferred by...are grown in the yolk sac of chicken embryos. The associated diseases are transmitted to humans via arthropod vectors such as fleas, ticks, lice

  12. DETERMINATION OF AMINOGLYCOSIDES IN FOOD BY FLUORESCENCE POLARIZATION IMMUNOASSAY

    OpenAIRE

    FARAFONOVA O.V.; Eremin, S. A.; ERMOLAEVA T.N.; VASILIEV S.V.

    2015-01-01

    The methodic for quantitative determination of aminoglycoside antibiotics (gentamicin, kanamycin, streptomycin, amikacin, neomycin) in food by polarization fluorescent immunoassay (FPIA) is developed. The size and structure influence of a fluorescent molecule on a fluorescence polarization degree is analyzed. Affinity constants of antibodies to compounds and tracers were estimated, optimized working concentration of tracers and antibodies that provide the maximum value of analytical signal. M...

  13. The effect of antibiotic therapy on the faecal excretion of Salmonella typhimurium by experimentally infected chickens.

    Science.gov (United States)

    Smith, H W; Tucker, J F

    1975-10-01

    Chickens in groups of 40 were infected orally with a nalidixic acid-resistant mutant of Salmonella typhimurium and then fed continuously on diets containing ampicillin, chloramphenicol, furazolidone, neomycin, oxytetracycline, polymixin, spectinomycin, streptomycin or a mixture of trimethoprim and sulphadiazine. The amount of S. typhimurium excreted in their faeces was estimated at intervals by culture on brilliant green agar containing sodium nalidixate, both direct and after enrichment in selenite broth; the amount of Escherichia coli excreted was estimated by culture on MacConkey agar. The feeding of diets containing 500 mg./kg. of ampicillin, furazolidone, neomycin, polymixin, spectinomycin or streptomycin or 100 mg./kg. of trimethoprim and 500 mg./kg. of sulphadiazine for 46 days reduced to a varying degree the amount of S. typhimurium and E. coli excreted, the greatest reduction in S. typhimurium being brought about by the last treatment. The effect was less obvious when the concentration of the antibiotics in the food was decreased fivefold. An important reason for the very limited effect of some of the antibiotics was the emergence of antibiotic-resistant populations of S. typhimurium and E. coli. High concentrations of antibiotic-resistant organisms also arose in the faeces of the chickens fed diets containing tetracyclines and chloramphenicol, treatments which had no apparent effect on the amount of S. typhimurium and E. coli excreted. Much of the antibiotic resistance encountered was determined by R factors, a particular R factor usually being found in the E. coli populations of individual chickens before it was found in their S. typhimurium populations. No S. typhimurium or E. coli were isolated that possessed R factors determining resistance to polymixin, furazolidone or trimethoprim. No S. typhimurium or E. coli were isolated that were polymixin-resistant and no S. typhimurium that were furazolidone-resistant. The few trimethoprim-resistant S

  14. Spontaneous kanamycin-resistant Escherichia coli mutant with altered periplasmic oligopeptide permease protein (OppA and impermeability to aminoglycosides Mutante espontâneo de Escherichia coli resistente à canamicina com expressão de oligopeptídeo permease periplasmática (OppA alterada e impermeabilidade aos aminoglicosídeos

    Directory of Open Access Journals (Sweden)

    Mônica B. Rodriguez

    1999-04-01

    Full Text Available A spontaneous kanamycin-resistant Escherichia coli mutant, showing cross resistance to five other aminoglycosides and absence of the OppA protein was isolated. [3H]-dihydrostreptomycin uptake is reduced in this mutant, implying that the oligopeptide transport system is involved in accumulation of aminoglycosides, although apparently not related with aminoglycoside permeability alteration due to bacterial adaptation to osmotic changes.Um mutante espontâneo de Escherichia coli foi selecionado com canamicina e mostrou resistência cruzada a cinco outros aminoglicosídeos e ausência da proteína OppA. A incorporação de diidroestreptomicina tritiada mostrou-se reduzida nesse mutante, implicando que o sistema de transporte de oligopeptídeos está envolvido na acumulação de aminoglicosídeos, embora aparentemente não esteja relacionado com a alteração de permeabilidade aos aminoglicosídeos decorrente da adaptação bacteriana a mudanças osmóticas.

  15. Scar formation in mice deafened with kanamycin and furosemide

    NARCIS (Netherlands)

    Zak, Magdalena; van der Linden, Cynthia A; Bezdjian, Aren; Hendriksen, Ferry G; Klis, Sjaak F L; Grolman, Wilko

    2016-01-01

    In mammals, hair cell loss is irreversible and leads to hearing loss. To develop and test the functioning of different strategies aiming at hair cell regeneration, animal models of sensorineural hearing loss are essential. Although cochleae of these animals should lack hair cells, supporting cells s

  16. Scar formation in mice deafened with kanamycin and furosemide

    NARCIS (Netherlands)

    Zak, Magdalena; van der Linden, Cynthia A; Bezdjian, Aren; Hendriksen, Ferry G; Klis, Sjaak F L; Grolman, Wilko

    In mammals, hair cell loss is irreversible and leads to hearing loss. To develop and test the functioning of different strategies aiming at hair cell regeneration, animal models of sensorineural hearing loss are essential. Although cochleae of these animals should lack hair cells, supporting cells

  17. HPLC同时测定复方硫酸新霉素药膜中盐酸丁卡因和克霉唑的含量%Simultaneous Determination of Tetracaine Hydrochloride and Clotrimazole in Compound Neomycin Sulfate Pellicles by HPLC

    Institute of Scientific and Technical Information of China (English)

    叶婷婷; 林珠

    2012-01-01

    OBJECTIVE To establish an HPLC method for simultaneous determination of tetracaine hydrochloride and clotrimazole in compound neomycin sulfate pellicles. METHODS An HPLC method was adopted. The column was Agilent HC-C18, the mobile phase was a mixture of methanol-0.1% triethylamine solution(adjusled to pH 3.0 with phosphoric acid) (85 : 15), the detection wavelength was 311 nm and 227 nm for tetracaine hydrochloride and clotrimazole, respectively, the flow rate was 0.8 mL·min‐1, the column temperature was maintained at 20 ℃, and the injection volume was 20 μL. RESULTS The linear range of tetracaine hydrochloride was 2.5-50.0 mg·L‐1(r=0.999 5), and the average recovery was 99.3%, RSD=0. 99% (n=9). The linear range of clotrimazole was 5.33-106.5 mg·L‐l(r=0.999 5), and the average recovery was 99.8%, RSD= 1.28% (n=9). CONCLUSION The method is simple, accurate, reproducible, and suitable for the simultaneous determination of tetracaine hydrochloride and clotrimazole in compound neomycin sulfate pellicles.%目的 建立同时测定复方硫酸新霉素药膜中盐酸丁卡因和克霉唑含量的方法.方法 采用HPLC,以Agilent HC-C18为色谱柱,流动相为甲醇-0.1%三乙胺溶液(用磷酸调pH至3.0)(85∶15),盐酸丁卡因和克霉唑的检测波长分别为311nm 和227nm,流速0.8mL·min-1,柱温20℃,进样20μL.结果 盐酸丁卡因在2.5~50.0mg·L-1内,峰面积与浓度呈良好的线性关系(r=0.9995),平均回收率为99.3%,RSD=0.99%(n=9);克霉唑在5.33~106.5mg·L-1内峰面积与浓度呈良好的线性关系(r=0.9995),平均回收率为99.8%,RSD=1.28%(n=9).结论 本法简便、准确、重复性好,可用于复方硫酸新霉素药膜中盐酸丁卡因和克霉唑的含量测定.

  18. Simultaneous Determination of Neomycin Sulfate and Hydrochloric Dyclonine in Compound Twain Oint-ments by HPLC-DAD-ELSD%HPLC-DAD-ELSD法同时测定复方吐温软膏中硫酸新霉素与盐酸达克罗宁的含量

    Institute of Scientific and Technical Information of China (English)

    段松冷; 曾蔚欣; 孙路路

    2015-01-01

    目的::建立同时测定复方吐温软膏中硫酸新霉素与盐酸达克罗宁含量的HPLC-DAD-ELSD法。方法:采用 Agilent ZOR BAXSB-C18色谱柱(250 mm ×4.6 mm,5μm),以含0.1 mol·L-1三氟乙酸的水溶液、乙腈为流动相,梯度洗脱,流速为1.0 ml·min-1,DAD的检测波长为282 nm,ELSD的漂移管温度为50℃,雾化温度为60℃,载气流速为1.6 L·min-1,柱温为35℃。结果:硫酸新霉素检测浓度的线性范围为141.540~323.520μg·ml-1(r=0.9996),平均回收率为98.87%,RSD=0.95%(n=9);盐酸达克罗宁检测浓的线性范围为28.000~64.000μg·ml-1(r=0.9996),平均回收率为99.57%,RSD=1.10%(n=9)。结论:该方法结果准确、灵敏度高、重复性好,在同一色谱条件下实现了复方吐温软膏中全部有效成分的含量测定,为其质量控制提供了依据。%Objective:To establish an HPLC-DAD-ELSD method for the simultaneous determination of neomycin sulfate and hy-drochloric dyclonine in compound Twaln ointments. Methods:The assay was performed on an Agilent ZOR BAXSB-C18 column(250 mm × 4. 6 mm, 5μm) with acetonitrile-water as the mobile phase with gradient elution at a flow rate of 1. 0 ml·min-1 . The detection wavelength of DAD was 282 nm. The evaporator temperature of ELSD was set at 50℃ and the nebulizer temperature was set at 60℃with the gas flow rate of 1. 6 L·min-1 . The column temperature was kept at 35℃. Results:The linear range of neomycin sulfate was 141. 54-323. 52 μg·mL-1(r=0. 999 6) with the average recovery of 98. 87%(RSD=0. 95%, n=9). The linear range of hydro-chloric dyclonine was 28. 00-64. 00 μg·mL-1(r=0. 999 6) with the average recovery of 99. 57%(RSD=1. 10%, n=9). Conclu-sion:The method is accurate, sensitive and reproducible, and under the same chromatographic conditions, the determination of all the active ingredients in compound Twaln ointments is achieved, which provides basis for the quality control.

  19. Agrobacterium tumefaciens-mediated GUS gene transfer to Sophora japonica L.

    Institute of Scientific and Technical Information of China (English)

    Zhang Xiao-ying; Wang Hua-fang; Yin Wei-lun; Zhu Zhen

    2006-01-01

    Agrobacterium-mediated genetic transformation of Sophorajaponica was standardized using the Agrobacterium tumefaciens strain LBA4404 that harbored the binary vector pBI121 containing genes for β-glucuronidase (GUS) and neomycin phosphotransterase (npt Ⅱ). S. japonica transformants were selected by the ability of the leaf explants to produce kanamycin-resistant calli that regenerated into kanamycin-resistant plantlets. Successful transformation was confirmed by histochemical assay for GUS activity, PCR analysis and Southern blot. The period of nearly two months was required for the regeneration of transgenic plantlets from the explants. The transformed plants resembled their parents in morphology.

  20. Generation of Transgenic Mice for Hygromycin and Neomycin Resistance Genes and Studies on Transgene Expression%潮霉素和新霉素抗性基因转基因小鼠的建立及其表达研究

    Institute of Scientific and Technical Information of China (English)

    党素英; 麻孙恺; 孙霞; 严兰珍; 王铸钢

    2005-01-01

    建立潮霉素(hygromycin,hyg)和新霉素(neomycin,neo)抗性基因转基因小鼠系,以获得两种抗性基因同时表达的小鼠胚胎成纤维细胞(MEFs),为条件性基因剔除或小鼠胚胎干细胞克隆的筛选创造条件.将pTK-hygR-pA,PGK-neoR-pA两个转录单元分别克隆至pBluescript载体中获得hygR-neoR双抗性基因表达质粒,以Kpn Ⅰ和Xba Ⅰ双酶切,回收4245bp串联基因片段,经显微注射获得HygR-neoR转基因小鼠.PCR及Southern blot鉴定转基因小鼠基因型;RT-PCR检测hygR、neoR基因在转基因阳性小鼠组织及MEFs中的表达.经显微注射共获得7只转基因阳性小鼠(Founders,G0代),经交配繁育,建立了6个转基因小鼠系.RT-PCR检测其中5个转基因阳性小鼠系F1代杂合子成年雌性小鼠肝脏、卵巢组织中hygR、neoR基因的表达,发现hn30,hn33,hn66和hn67系阳性小鼠的1种或2种组织中有hygR、neoR的表达;RT-PCR检测发现hyg和neo抗性基因在从h30和hn66系分离培养的MEFs中表达.通过显微注射,建立了表达hyg-neo抗性基因的转基因小鼠系,转基因表达检测发现两个转基因小鼠系的胚胎成纤维细胞中表达双抗性基因.

  1. DETERMINATION OF AMINOGLYCOSIDES IN FOOD BY FLUORESCENCE POLARIZATION IMMUNOASSAY

    Directory of Open Access Journals (Sweden)

    FARAFONOVA O.V.

    2015-01-01

    Full Text Available The methodic for quantitative determination of aminoglycoside antibiotics (gentamicin, kanamycin, streptomycin, amikacin, neomycin in food by polarization fluorescent immunoassay (FPIA is developed. The size and structure influence of a fluorescent molecule on a fluorescence polarization degree is analyzed. Affinity constants of antibodies to compounds and tracers were estimated, optimized working concentration of tracers and antibodies that provide the maximum value of analytical signal. Methods were tested in the antibiotics identification in milk, eggs and chicken.

  2. Antibiotic resistance of Staphylococcus aureus strains isolated from fish processing factory workers

    OpenAIRE

    1988-01-01

    One hundred and twenty two strains of Staphylococcus aureus isolated from throats and palms of 39 workers from 6 fish processing factories situated in and around Cochin were tested for their sensitivity to nine commonly used antibiotics-ampicillin, chloramphenicol, erythromycin, kanamycin, neomycin, penicillin, polymyxin-B, streptomycin and tetracycline. Highest percentage of resistance was observed towards ampicillin followed by penicillin i.e. 64.75% and 59.84%. Resistance towards other ant...

  3. Host-dependent transposon Tn5-mediated streptomycin resistance.

    OpenAIRE

    1984-01-01

    Transposon Tn5 encodes streptomycin resistance in addition to kanamycin-neomycin resistance. This resistance was not detectable in Escherichia coli but was efficiently expressed in Rhizobium meliloti and certain other strains. By analysis of cloned Tn5 restriction endonuclease fragments, the streptomycin resistance (str) gene was located in the right-hand side of the central region as the transposon is conventionally drawn. Transcription of str appeared to originate at pL, the promoter for th...

  4. Complexation of anionic copolymers of acrylamide and N-(2-hydroxypropyl)methacrylamide with aminoglycoside antibiotics

    Science.gov (United States)

    Solovskii, M. V.; Tarabukina, E. B.; Amirova, A. I.; Zakharova, N. V.; Smirnova, M. Yu.; Gavrilova, I. I.

    2014-03-01

    The complexation of aminoglycoside antibiotics neomycin, gentamicin, kanamycin, and amikacin in the form of free bases with carboxyl- and sulfo-containing copolymers of acrylamide and N-(2-hydroxypropyl)methacrylamide (HPMA) in water and water-salt solutions is studied by means of viscometry, equilibrium dialysis, potentiometric titration, and molecular hydrodynamics. Factors influencing the stability of formed copolymer-antibiotic complexes and determinations of their toxicity are established.

  5. Multiple antibiotic resistance among gram negative bacteria isolated from poultry.

    Science.gov (United States)

    Ansari, F A; Khatoon, H

    1994-03-01

    Gram negative bacteria, including species of Salmonella, Escherichia, Pseudomonas and Klebsiella, isolated from poultry, were screened for their resistance to the commonly used antibiotics: ampicillin, chloramphenicol, gentamycin, kanamycin, neomycin, polymyxin B, streptomycin and tetracycline. Of the 500 bacteria screened, 351 were found to be resistant to one or more antibiotics at the level of 50 micrograms/ml. Various patterns of antibiotic resistance observed during these studies have been reported.

  6. Antibiotic resistance in Salmonella Enteritidis isolated from broiler carcasses Resistência antimicrobiana em Salmonella Enteritidis isoladas de carcaças de frango

    OpenAIRE

    Martha Oliveira Cardoso; Aldemir Reginato Ribeiro; Luciana Ruschel dos Santos; Fernando Pilotto; Moraes,Hamilton L.S.; Carlos Tadeu Pippi Salle; Silvio Luís da Silveira Rocha; Vladimir Pinheiro do Nascimento

    2006-01-01

    Eighty Salmonella Enteritidis strains isolated from broiler carcasses between May 1995 and April 1996 in the State of Rio Grande do Sul, Brazil, were tested for antibiotic susceptibility using the disk diffusion method. Resistance to colistin, novobiocin, erythromycin and tetracycline was observed in 100% of the isolates. The strains showed intermediate resistance at different levels to kanamycin (1.25%), enrofloxacin (3.75%), neomycin (3.75%), fosfomycin (20%), sulphonamides (86.25%) and nit...

  7. 21 CFR 558.455 - Oxytetracycline and neomycin.

    Science.gov (United States)

    2010-04-01

    ... of disease symptoms. A withdrawal period has not been established for use in preruminating calves. Do... beyond remission of disease symptoms. Withdraw 5 d before slaughter. 048164066104 (iv) To provide 25 mg... Chickens: For control of infectious synovitis caused by Mycoplasma synoviae; control of fowl cholera caused...

  8. Detection of kanamycin resistant gene expression in vector-transfected mammalian cells and milk samples of vector-injected cow%自身启动子控制的卡那霉素抗性基因在哺乳动物细胞中表达的检测

    Institute of Scientific and Technical Information of China (English)

    翁玉根; 吉玉辉; 孙怀昌; 张鑫宇; 朱建国; 易明梅; 袁耀明

    2012-01-01

    To investigate whether the kanamycin resistant (KanR) gene in mammary-specific expression vector p215C3LYZ for treating bovine mastitis could express active protein in mammalian cells under the control of its own promoter, KanR gene was amplified by PCR using mammary-specific expression vector p215C3LYZ as the template and subcloned into prokaryotic expression vector pQE-31. After transformation and cultivation on Kan-containing agar plates, Kan-resistant E. coll colonies were obtained and an expected recombinant protein was expressed after IPTG induction. After separation on SDS-PAGE, the protein band was excised and specific antiserum was obtained by immunization of mice for 6 times. The specificity of the antiserum was confirmed using Western blotting. The prokaryotic expression vector pQE-Kan and eukaryotic expression vector p215C3LYZ, both of which contained KanR gene, were transfected into COS-1 cells and the supernatants were collected after cultivation in antibiotic-free medium, DH5a E. c.oli were inoculated into the supernatants in the absence or presence of Kan and the OD600 values were detected after 24h cultivation. The results showed that growth of the indicator bacterium was inhibited signifi-cantly in the presence of Kan, indicating that the Kan resistant protein was not expressed in the supernatants of the vector-transfected cells. Western blotting of the cell lysates confirmed that Kan resistant protein was not expressed in the vector-transfected cells. In addition, no Kan resistant protein was detected in concentrated milk samples from cows after intramammary injection with vector p215C3LYZ. These experimental data demonstrate that the promoter of KanR gene is inactive in mammalian cells, providing additional safety evidence using the p215C3LYZ vector for treating bovine mastitis.%为了研究卡那霉素抗性(KanR)基因能否在哺乳动物细胞中表达以及用含相同抗性基因重组质粒防治奶牛乳腺炎的

  9. 腺伴随病毒介导 ADNF-9基因转染对豚鼠卡那霉素致聋的治疗作用%Treatment of adeno-associated virus mediated ADNF-9 gene transfection on guinea pig cochlea deafened by kanamycin

    Institute of Scientific and Technical Information of China (English)

    景阳; 郑国玺; 祝康; 刘晖; 张文

    2015-01-01

    Objective:To observe the effect of rAAV-NT4-ADNF-9 in treating ototoxicity associated with aminoglycoside antibiotics.Methods:The guinea pigs were used,and its auvicle reflex and auditory brain-stem response(ABR)were normal.The ototoxic animal model of guinea pigs were made by the method of hypodemic injection of Kanamycin(400 mg/kg).In animals under general anesthesia,the guinea pigs were cut off the skin of ringt ear sulcus posterior and opened acoustic capsule,then injected the rAAV-NT4-AD-NF-9 10 μL from fossula rotunda.After 14 days,the right acoustic capsule were dislodged.The cochlear were stained with AgNO3 .The cochlear hair cells were observed by contrast phase microscope.Results:Taking the group for rAAV-NT4-ADNF-9 as example,the threshold ABR decreased remarkably after treat-ment(P <0.05).Through the phase contrast microscope the cochlear basilar membrane was complete and the outer hair cells had dot deletion.The thresholds of ABR had no notable differences for the other three groups before and after treatment.Taking comparison between the artificial endolymph control group and empty vector group,through the phase contrast microscope after treatment,hair cells showed a patchy loss in the former group and there was no such phenomenon in the latter group.The average number of hair cells observed in the group of ADNF was significantly higher than that of the artificial endolymph control group and empty vector group(P <0.05).Conclusion:rAAV-NT4-ADNF-9 is able to be transfected into cochle-ar,and to express secretory NT4-ADNF-9 peptide,which preventing hair cells from impairment.%目的:观察包装重组的腺伴随病毒(adeno-associated virus,AAV)rAAV-NT4-ADNF-9对卡拉霉素致聋豚鼠的治疗作用。方法:选用耳廓反射和听性脑干反应(auditory brainstem response,ABR)正常的白色红目豚鼠,在大腿内侧肌肉注射卡那霉素400 mg/(kg·d),连续注射6 d,制备耳聋模型。在全身麻醉的条件

  10. Environmental and genetic factors affecting mutability to aminoglycoside antibiotics among Escherichia coli K12 strains

    Directory of Open Access Journals (Sweden)

    Monteiro A.C.M.

    2003-01-01

    Full Text Available Environmental and genetic factors affecting the in vitro spontaneous mutation frequencies to aminoglycoside resistance in Escherichia coli K12 were investigated. Spontaneous mutation frequencies to kanamycin resistance were at least 100 fold higher on modified Luria agar (L2 plates, when compared to results obtained in experiments carried out with Nutrient agar (NA plates. In contrast to rifampincin, the increased mutability to kanamycin resistance could not be attributed to a mutator phenotype expressed by DNA repair defective strains. Kanamycin mutant selection windows and mutant preventive concentrations on L2 plates were at least fourfold higher than on NA plates, further demonstrating the role of growth medium composition on the mutability to aminoglycosides. Mutability to kanamycin resistance was increased following addition of sorbitol, suggesting that osmolarity is involved on the spontaneous mutability of E. coli K12 strains to aminoglycosides. The spontaneous mutation rates to kanamycin resistance on both L2 and NA plates were strictly associated with the selective antibiotic concentrations. Moreover, mutants selected at different antibiotic concentrations expressed heterogeneous resistance levels to kanamycin and most of them expressing multiple resistance to all tested aminoglycoside antibiotics (gentamicin, neomycin, amykacin and tobramycin. These results will contribute to a better understanding of the complex nature of aminoglycoside resistance and the emergence of spontaneous resistant mutants among E. coli K12 strains.

  11. Aminoglycoside resistance in Haemophilus influenzae.

    Science.gov (United States)

    Gomez-Lus, R; Vergara, Y

    1995-04-01

    From September 1, 1990 to December 31, 1993 a total of 425 Haemophilus influenzae strains from clinical specimens were isolated in the Microbiology Laboratory of the Zaragoza University Hospital. Of these strains, 16 (33.33%) were resistant to kanamycin, neomycin, paromomycin, lividomycin and streptomycin. Demonstration of APH (3')-I activity by the phosphocellulose paper binding assay, based on the incorporation of radiolabel into lividomycin was sixfold greater than into butirosin. Two DNA probes were prepared to screen for the genes encoding APH(3') activity in kanamycin-resistant H. influenzae. Homology was observed between the aphA1 DNA probe and total cellular DNA from all 16 APH(3')-I producers. On the other hand, streptomycin-resistance was not through metabolic modification of the antibiotic.

  12. Agrobacterium-mediated transformation of Ruta graveolens L.

    Science.gov (United States)

    Lièvre, Karine; Tran, Thi Lê Minh; Doerper, Sébastien; Hehn, Alain; Lacoste, Paul; Thomasset, Brigitte; Bourgaud, Frédéric; Gontier, Eric

    2009-01-01

    Agrobacterium tumefaciens is used to develop a genetic transformation method for a medicinal plant Ruta graveolens. The direct plant regeneration strategy is preferred to callus line establishment. In vitro seedlings, 2- -to 3-wk-old, are used to excise hypocotyls and co-cultivated for 3 d with A. tumefaciens strain C58C1Rif containing plasmid pTDE4 harbouring neomycin phosphotransferase (npt II, kanamycin resistance) and beta-glucuronidase encoding genes. The Southern blot analysis has shown that 78% kanamycin resistant plants contain gene encoding beta-glucuronidase. The GUS histochemical assay shows that 67% transgenic plants exhibit the corresponding enzymatic activity. Routine transformation efficiency of R. graveolens L. is 11% and could reach up to 22%. Transgenic plants are grown in the greenhouse within 4 months after the initial seedlings.

  13. Studies on Genetic Transformatiom of Embryogenic Suspension Cultures of Sweetpotato

    Institute of Scientific and Technical Information of China (English)

    ZHAI Hong; LIU Qing-chang

    2003-01-01

    Genetic transformation of embryogenic suspension cultures of sweetpotato cv. Lizixiang wasconducted by using Agrobacterium tumefaciens strain A208SE harboring the binary vectors pROA93 with β-glucronidase (GUS) and neomycin phosphotransferase (NPT Ⅱ ) genes. The results indicated that embryogenicsuspension cultures precultured for 1 -3 d were suitable for the transformation. The optimal cocultivation timewas 4 - 5 d. The optimal concentration of kanamycin was 50-75 mg L-1 for suspension culture and 100 mg L-1for embryogenic callus proliferation and plant regeneration. The optimal concentration of carbencillin was 100mg L-1. Transgenic plants identified with GUS assays and PCR analyses were obtained.

  14. Instability of multiple drug resistance plasmids in Salmonella typhimurium isolated from poultry.

    OpenAIRE

    Brown, D J; Threlfall, E. J.; Rowe, B

    1991-01-01

    Plasmids in five strains of Salmonella typhimurium resistant to ampicillin, chloramphenicol, gentamicin, neomycin/kanamycin, streptomycin, sulphonamides, tetracyclines and trimethoprim (ACGKSSuTTm), CGKSSuTTm, ACSSuT or CSSuT which had been isolated from poultry in the first 3 months of 1989 have been characterized and compared with plasmids in two strains of R-types ACGKSSuTTm and ASSuTTm isolated from two patients later in the year. With the exception of the human isolate of R-type ASSuTTm,...

  15. [Drug resistance of Escherichia coli strains isolated from poultry].

    Science.gov (United States)

    Giurov, B; Korudzhiĭski, N; Bineva, I

    1981-01-01

    Studied was the sensitivity of a total of 143 strains of Escherichia coli, isolated from young birds and broilers died from coli septicaemia, to antibiotics and chemotherapeutics. The following descending order was established: gentamycin, carbenicillin, ampicillin, furazolidon, borgal, kanamycin, strep tomycin, chloramphenicol, neomycin sulphathiazole, and tetracycline. Markers of resistance were established with all strains with regard to the therapeutic agents in current and prospective use in industrial poultry farming. It is stated that a preliminary antibiogram is indispensable in order to obtain dependable results in the treatment of animals affected with colibacteriosis. An alternative is to apply directly those drugs to which the strains have shown highest sensitivity.

  16. aph(3′)-IIb, a Gene Encoding an Aminoglycoside-Modifying Enzyme, Is under the Positive Control of Surrogate Regulator HpaA

    OpenAIRE

    Zeng, Lin; Jin, Shouguang

    2003-01-01

    Pseudomonas aeruginosa harbors a chromosomal aminoglycoside phosphotransferase gene, aph(3′)-IIb, which confers P. aeruginosa resistance to several important aminoglycoside antibiotics, including kanamycin A and B, neomycin B and C, butirosin, and seldomycin F5. The aph(3′)-IIb gene has been found to be regulated by an AraC-type transcriptional regulator (HpaA) encoded by a gene located upstream of the aph(3′)-IIb gene. In the presence of 4-hydroxyphenylacetic acid (4-HPA), HpaA activates the...

  17. Deciphering the details of RNA aminoglycoside interactions: from atomistic models to biotechnological applications

    Energy Technology Data Exchange (ETDEWEB)

    Ilgu, Muslum [Iowa State Univ., Ames, IA (United States)

    2012-01-01

    A detailed study was done of the neomycin-B RNA aptamer for determining its selectivity and binding ability to both neomycin– and kanamycin-class aminoglycosides. A novel method to increase drug concentrations in cells for more efficiently killing is described. To test the method, a bacterial model system was adopted and several small RNA molecules interacting with aminoglycosides were cloned downstream of T7 RNA polymerase promoter in an expression vector. Then, the growth analysis of E. coli expressing aptamers was observed for 12-hour period. Our analysis indicated that aptamers helped to increase the intracellular concentration of aminoglycosides thereby increasing their efficacy.

  18. Multiple keys for a single lock: the unusual structural plasticity of the nucleotidyltransferase (4')/kanamycin complex.

    Science.gov (United States)

    Matesanz, Ruth; Diaz, José Fernando; Corzana, Francisco; Santana, Andrés G; Bastida, Agatha; Asensio, Juan Luis

    2012-03-05

    The most common mode of bacterial resistance to aminoglycoside antibiotics is the enzyme-catalysed chemical modification of the drug. Over the last two decades, significant efforts in medicinal chemistry have been focused on the design of non- inactivable antibiotics. Unfortunately, this strategy has met with limited success on account of the remarkably wide substrate specificity of aminoglycoside-modifying enzymes. To understand the mechanisms behind substrate promiscuity, we have performed a comprehensive experimental and theoretical analysis of the molecular-recognition processes that lead to antibiotic inactivation by Staphylococcus aureus nucleotidyltransferase 4'(ANT(4')), a clinically relevant protein. According to our results, the ability of this enzyme to inactivate structurally diverse polycationic molecules relies on three specific features of the catalytic region. First, the dominant role of electrostatics in aminoglycoside recognition, in combination with the significant extension of the enzyme anionic regions, confers to the protein/antibiotic complex a highly dynamic character. The motion deduced for the bound antibiotic seem to be essential for the enzyme action and probably provide a mechanism to explore alternative drug inactivation modes. Second, the nucleotide recognition is exclusively mediated by the inorganic fragment. In fact, even inorganic triphosphate can be employed as a substrate. Third, ANT(4') seems to be equipped with a duplicated basic catalyst that is able to promote drug inactivation through different reactive geometries. This particular combination of features explains the enzyme versatility and renders the design of non-inactivable derivatives a challenging task.

  19. 21 CFR 524.1204 - Kanamycin sulfate, calcium amphomycin, and hydrocortisone acetate.

    Science.gov (United States)

    2010-04-01

    ... antibiotics: Acute otitis externa, furunculosis, folliculitis, pruritus, anal gland infections, erythema, decubital ulcer, superficial wounds, and superficial abscesses. (2) The ointment should be applied to the... of the anal gland, the drug should be introduced into the orifice of the gland and not through...

  20. Highly sensitive spectrofluorimetric method for determination of certain aminoglycosides in pharmaceutical formulations and human plasma.

    Science.gov (United States)

    Omar, Mahmoud A; Nagy, Dalia M; Hammad, Mohamed A; Aly, Alshymaa A

    2013-06-01

    A simple, reliable, highly sensitive and selective spectrofluorimetric method has been developed for determination of certain aminoglycosides namely amikacin sulfate, tobramycin, neomycin sulfate, gentamicin sulfate, kanamycin sulfate and streptomycin sulfate. The method is based on the formation of a charge transfer complexes between these drugs and safranin in buffer solution of pH 8. The formed complexes were quantitatively extracted with chloroform under the optimized experimental conditions. These complexes showed an excitation maxima at 519-524 nm and emission maxima at 545-570 nm. The calibration plots were constructed over the range of 4-60 pg mL(-1) for amikacin, 4-50 pg mL(-1) for gentamicin, neomycin and kanamycin, 4-40 pg mL(-1) for streptomycin and 5-50 pg mL(-1) for tobramycin. The proposed method was successfully applied to the analysis of the cited drugs in dosage forms. The proposed method was validated according to ICH and USP guidelines with respect to specificity, linearity, accuracy, precision and robustness. The high sensitivity of the proposed method allowed determination of amikacin and gentamicin in spiked and real human plasma.

  1. Antibacterial activity of amphiphilic tobramycin.

    Science.gov (United States)

    Dhondikubeer, Ramesh; Bera, Smritilekha; Zhanel, George G; Schweizer, Frank

    2012-10-01

    Amphiphilic aminoglycoside antimicrobials are an emerging class of new antibacterial agents with novel modes of action. Previous studies have shown that amphiphilic neomycin-B and kanamycin-A analogs restore potent antibacterial activity against Gram-positive neomycin-B- and kanamycin-A-resistant organisms. In this paper, we investigated the antibacterial properties of a series of amphiphilic tobramycin analogs. We prepared tobramycin-lipid conjugates, as well as tobramycin-peptide triazole conjugates, and studied their antibacterial activities against a panel of Gram-positive and Gram-negative bacterial strains, including isolates obtained from Canadian hospitals. Our results demonstrate that the antibacterial activity of amphiphilic tobramycin is greatly affected by the length and nature of the hydrophobic lipid tail, whereas the nature of the polycationic headgroup or the number of cationic charges appear to be less important. Replacement of the hydrophobic tail by a fluorinated lipid confers good activity against two Pseudomonas strains and reduces hemolytic activity. However, susceptibility studies in the presence of bovine serum albumin indicate that all amphiphilic tobramycin analogs are strongly protein-bound, leading to a typical four- to eight-fold increase in MIC.

  2. Temporal bone studies of the human peripheral vestibular system. Aminoglycoside ototoxicity.

    Science.gov (United States)

    Tsuji, K; Velázquez-Villaseñor, L; Rauch, S D; Glynn, R J; Wall, C; Merchant, S N

    2000-05-01

    Quantitative assessments of vestibular hair cells and Scarpa's ganglion cells were performed on 17 temporal bones from 10 individuals who had well-documented clinical evidence of aminoglycoside ototoxicity (streptomycin, kanamycin, and neomycin). Assessment of vestibular hair cells was performed by Nomarski (differential interference contrast) microscopy. Hair cell counts were expressed as densities (number of cells per 0.01 mm2 surface area of the sensory epithelium). The results were compared with age-matched normal data. Streptomycin caused a significant loss of both type I and type II hair cells in all 5 vestibular sense organs. In comparing the ototoxic effect on type I versus type II hair cells, there was greater type I hair cell loss for all 3 cristae, but not for the maculae. The vestibular ototoxic effects of kanamycin appeared to be similar to those of streptomycin, but the small sample size precluded definitive conclusions from being made. Neomycin did not cause loss of vestibular hair cells. Within the limits of this study (maximum postototoxicity survival time of 12 months), there was no significant loss of Scarpa's ganglion cells for any of the 3 drugs. The findings have implications in several clinical areas, including the correlation of vestibular test results to pathological findings, the rehabilitation of patients with vestibular ototoxicity, the use of aminoglycosides to treat Meniere's disease, and the development of a vestibular prosthesis.

  3. Aminoglycosides resistance in clinical isolates of Staphylococcus aureus from a University Hospital in Bialystok, Poland.

    Directory of Open Access Journals (Sweden)

    Katarzyna Kaczyńska

    2008-06-01

    Full Text Available Staphylococcus aureus obtained from a University Hospital in Poland were characterized in relation to resistance to aminoglycoside antibiotics and the distribution of the genes encoding the most clinically relevant aminoglycoside modifying enzymes (AMEs. Of a total of 118 S. aureus, 45 (38.1% isolates were found to be resistant to at least one of the tested antibiotics. All aminoglycoside resistant isolates except one 44 (97.8% were resistant to kanamycin. The majority of strains 37 (82.2% and 32 (71.1% expressed resistance to neomycin and tobramycin, respectively. Eleven strains (24.4% were resistant to gentamicin or amikacin. All S. aureus strains were sensitive to netilmicin. The most prevalent resistance gene was aac(6'-Ie+aph(2' found in 13 (28.9% strains and 12 (26.7% isolates carried ant(4'-Ia gene, whilst aph(3'-IIIa gene was detected in only 7 (15.6% isolates. Additionally, the ant(6-Ia and str genes were detected in 14 (31.1% and 2 (4.4% strains, respectively. Ten (22.2% strains resistant to amikacin, tobramycin, kanamycin or neomycin did not harbor any of the above-noted genes.

  4. Label-free and multiplex detection of antibiotic residues in milk using imaging surface plasmon resonance-based immunosensor.

    Science.gov (United States)

    Rebe Raz, Sabina; Bremer, Maria G E G; Haasnoot, Willem; Norde, Willem

    2009-09-15

    Monitoring of antimicrobial drug residues in foods relies greatly on the availability of adequate analytical techniques. Currently, there is a need for a high-throughput screening method with a broad-spectrum detection range. This paper describes the development of a microarray biosensor, based on an imaging surface plasmon resonance (iSPR) platform, for quantitative and simultaneous immunodetection of different antibiotic residues in milk. Model compounds from four major antibiotic families: aminoglycosides (Neomycin, Gentamicin, Kanamycin, and Streptomycin), sulfonamides (Sulfamethazine), fenicols (Chloramphenicol), and fluoroquinolones (Enrofloxacin) were detected using a single sensor chip. By multiplexing seven immunoassays in a competitive format, we were able to measure all the target compounds at parts per billion (ppb) levels in buffer and in 10x-diluted milk. The assays for Neomycin, Kanamycin, Streptomycin, Enrofloxacin, and Sulfamethazine were sensitive enough for milk control at maximum residue levels as established in the European Union. The overall performance of the biosensor was determined to be comparable to that of conventional four-channel surface plasmon resonance (SPR)-based biosensors, in terms of assay sensitivity and robustness. Combining the advantages of a SPR sensor and a microarray, utilization of the biosensor described here offers a promising alternative to the existing methods and is highly relevant for multianalyte food profiling.

  5. AXENIC CULTURE OF FREE-LIVING CONCHOCELIS OF PORPHYRA YEZOENSIS AND PORPHYRA HAITANENSIS

    Institute of Scientific and Technical Information of China (English)

    刘会莲; 帅莉; 段德麟; 徐怀恕

    2002-01-01

    After discarding marine microorganisms from conchocelis of Porphyra yezoensis and Porphyra haitanensis, their axenic cultures were obtained through treatment with antibiotics. Antibiotic disc tests were carried out to determine the effectiveness of each antibiotic in eliminating contaminating microorganisms. Five of 12 antibiotics tested were selected and used to produce the axenic cultures in this study, which showed that 200 μg/mL streptomycin, 250 μg/mL penicillin, 252 μg/mL kanamycin, 30 μg/mL neomycin, 200 μg/mL chloramphenicol were effective concentrations for eliminating microorganisms from conchocelis when antibiotics were added singly step by step; whereas simultaneous combination of 150 μg/mL streptomycin, 250 (or 350) μg/mL penicillin, 150 (or 250) μg/mL kanamycin, 70 μg/mL neomycin and 200 μg/mL chloramphenicol was also effective for producing the axenic cultures. However, it seemed that the treatments with antibiotics applied individually were more feasible than those with all antibiotics added at the same time. This may be due to the combined inhibiting effect of antibiotics on the growth and development of conchocelis.

  6. Structure of polysaccharide antibiotics; Structure des antibiotiques polysaccharidiques

    Energy Technology Data Exchange (ETDEWEB)

    Matutano, L. [Commissariat a l' Energie Atomique, Saclay (France). Centre d' Etudes Nucleaires

    1966-07-01

    Study of the structure of antibiotics having two or several sugars in their molecule. One may distinguish: the polysaccharide antibiotics themselves, made up of two or several sugars either with or without nitrogen, such as streptomycin, neomycins, paromomycine, kanamycin, chalcomycin; the hetero-polysaccharide antibiotics made up of one saccharide part linked to an aglycone of various type through a glucoside: macrolide, pigment, pyrimidine purine. Amongst these latter are: erythromycin, magnamycin, spiramycin, oleandomycin, cinerubin and amicetin. The sugars can either play a direct role in biochemical reactions or act as a dissolving agent, as far as the anti-microbe power of these antibiotics is concerned. (author) [French] Etude de la structure des antibiotiques qui possedent dans leur molecule deux ou plusieurs sucres. On distingue: les antibiotiques polysaccharidiques proprement dits, constitues de deux eu plusieurs sucres azotes ou non, tels que streptomycines, neomycines, paromomycines, kanamycine, chalcomycine; les antibiotiques heteropolysaccharidiques formes d'une partie saccharidique liee glycosidiquement a un aglycone de nature diverse: macrolide - pigment - purine pyrimidine. On compte parmi ceux-ci: erythroraycine rnagnamycine, spiromycine, oleandomycine, cinerubine et amicetine. Dans le pouvoir antimicrobien de ces antibiotiques les sucres peuvent jouer soit un role direct dans les reactions biochimiques, soit agir comme solubilisant. (auteur)

  7. The incidence of antibiotic resistance and other characteristics amongst Escherichia coli strains causing fatal infection in chickens: the utilization of these characteristics to study the epidemiology of the infection.

    Science.gov (United States)

    Heller, E D; Smith, H W

    1973-12-01

    Of 173 epidemiologically unrelated strains of Escherichia coli isolated from the pericardial sac of chickens that had died from infection with these organisms in England in 1972, approximately 1 year after the introduction of legislation forbidding the routine use of feeds containing ;therapeutic' antibiotics, 83.8% were resistant to sulphonamides, 31.2% to tetracyclines, 20.8% to furazolidone, 18.5% to streptomycin, 2.9% to spectinomycin and 1.2% to ampicillin; none of the strains were resistant to chloramphenicol, neomycin, polymixin, trimethoprim or nalidixic acid. The sulphonamide resistance and possibly some of the resistance to other agents might have been the consequence of sulphonamides being exempted from the legislation. Much of the resistance, with the exception of that to furazolidone, was of the transferable type. Many strains possessed transfer factors in the absence of any known transferable characteristic. Colicine production was twice as common in the pathogenic strains as in a collection of strains isolated from the faeces of healthy chickens; about half of it was transferable.By means of serology, antibiotic resistance and other markers, it was found that several different kinds of E. coli were usually incriminated in any one outbreak of E. coli infection in broiler chickens. Sometimes the same kinds of E. coli were found in outbreaks in consecutive crops of chickens on the same farm. New kinds, too, appeared to be brought in by replacement chickens.

  8. 21 CFR 520.1921 - Prochlorperazine, isopropamide, with neomycin sustained-release capsules.

    Science.gov (United States)

    2010-04-01

    ... sustained-release capsules. 520.1921 Section 520.1921 Food and Drugs FOOD AND DRUG ADMINISTRATION... capsules. (a) Specifications. Each capsule contains either: (1) Capsule No. 1: 3.33 milligrams of... base (as the sulfate); or (2) Capsule No. 3: 10 milligrams of prochlorperazine (as the dimaleate), 5...

  9. 21 CFR 524.1484h - Neomycin, penicillin, polymyxin, hydrocortisone suspension.

    Science.gov (United States)

    2010-04-01

    ... topical exposure to penicillin. (d) Conditions of use—dogs—(1) Amount. Rub a small amount into the... day. (2) Indications for use. Treatment of summer eczema, atopic dermatitis, interdigital eczema, and.... For use in dogs only. Shake drug thoroughly and clean lesion before using. If redness, irritation,...

  10. The effect of neomycin and streptomycin on the electrical polarisability of aqueous suspensions of Escherichia coli.

    Science.gov (United States)

    Morris, V J; Jennings, B R

    1975-06-12

    Aqueous suspensions of bacteria scatter light strongly. In addition, the bacteria exhibit strong induced dipole moments in an electric field. In this note we report how, by measuring the intensity of the scattered light, the electric polarisability (alpha) of Escherichia coli could be monitored as small quantities of antibiotics were added to the suspensions. The effect of the presence of quite small quantities of antibiotic on the electrical polarisability, which gave rise to the induced dipole, was dramatic. From the hypothesis that alpha had its origins in the bacteria-solvent interface, a theory is presented which adequately accounts for both alpha and its changes in the presence of these antibiotics. The study is taken to suggest that the antibiotic molecules were adsorbed on to the bacterial surface thereby reducing the surface charge. This in turn reduces the number of counterions and the apparent induced dipole moment. Because the electric-field scattering method is both quick and sensitive to changes in alpha, it may prove a valuable method for the study of antibiotic action on cell and microorganism surfaces.

  11. 21 CFR 524.1484c - Neomycin sulfate, isoflupredone acetate, tetracaine hydrochloride ointment.

    Science.gov (United States)

    2010-04-01

    ...) Conditions of use. (1) It is used in treating such conditions as acute otitis externa in dogs and to a lesser degree, chronic otitis externa in dogs. It also is effective in treating anal gland infections and moist..., following ear trimming and castrating operations. (2) In treatment of otitis externa and other...

  12. 21 CFR 524.1881b - Prednisolone acetate-neomycin sulfate sterile suspension.

    Science.gov (United States)

    2010-04-01

    ... conjunctivitis, acute otitis externa, and chronic otitis externa in dogs and cats. (2) For beginning treatment of... hour period. When improvement occurs, the dosage may be reduced to 1 drop 2 to 4 times daily. In otitis externa, 2 to 6 drops may be placed in the external ear canal 2 or 3 times daily. (3) All...

  13. 21 CFR 524.1484f - Neomycin sulfate, prednisolone acetate, tetracaine hydrochloride eardrops.

    Science.gov (United States)

    2010-04-01

    ... chapter. (c) Conditions of use. (1) It is useful in treating such conditions as acute otitis externa and, to a lesser degree, chronic otitis externa in dogs and cats. It is indicated as treatment or...-susceptible organisms and/or allergy. In otitis externa, 2 to 6 drops may be placed in the external ear...

  14. 21 CFR 524.1600b - Nystatin, neomycin, thiostrepton, and triamcinolone acetonide ophthalmic ointment.

    Science.gov (United States)

    2010-04-01

    ... conjunctivitis in cats and dogs and for infectious kerato-conjunctivitis (pink eye) in cattle. (2) It is to be administered as follows: (i) For conjunctivitis and keratitis: Apply one drop of ointment to the affected eye(s... infectious kerato-conjunctivitis: Apply small line of ointment to the affected eye(s) once daily....

  15. Study of Klebsiella pneumoniae producing extended-spectrum β-lactamases against aminoglycosides

    Institute of Scientific and Technical Information of China (English)

    WEI FENG SHI; SU JIAN WANG; JIAN PING QIN

    2007-01-01

    Klebsiella pneumoniae ( K. pneumoniae) is one of the main gram-negative bacilli in clinical practice. Nosocomial infections caused by K. pneumoniae producing extended-spectrum β-lactamases (ESBLs) are very difficult to treat. This paper investigated the resistant characteristics of K. pneumoniae producing ESBLs and their aminoglycoside-modifying enzyme gene expressions including Nacetyltransferases and O-adenyhransferases. Bacteria identification and ESBLs confirmatory tests were performed by Phoenix TM-100 system. And minimum inhibitory concentrations (MICs) of gentamicin,amikacin, kanamycin, tobramycin, netilmicin and neomycin in 53 K. pneumoniae isolates were detected by agar dilution. In addition, six aminoglycoside-modifying enzyme genes were amplified by polymerase chain reaction (PCR) and verified by DNA sequencer. It was found that imipenem and meropenem against 120 K. pneumoniae isolates produced powerful antimicrobial activities. The resistant rates of gentamicin and amikacin were 55.0% and 46.7%, respectively. Except neomycin,MIC50 and MIC90 of gentamicin, amikacin, kanamycin, tobramycin and netilmicin in 53 K. pneumoniae were all > 128 μg/ml, and the resistant rates were 83.0%, 52.3%, 75.5%, 81. 1% and 69.8%, respectively. However, neomycin was only 39.6%. In addition, five modifying enzyme genes, including aac(3)- Ⅰ , aac(3)-Ⅱ, aac(6′) - Ⅰ b, ant(3″) - Ⅰ, ant(2″) - Ⅰ genes, were found in 53 isoahes except aac (6′)-Ⅱ, and their positive rates were 11.3%, 67.9%, 47.2%,1.9 % and 39.6 %, respectively. It was also confirmed by nucleotide sequence analysis that the above resistant genes shared nearly 100% identities with GenBank published genes. The results obtained in the present study indicated that K. pneumoniae producing ESBLs strains are rapidly spreading in our hospital, and their resistance to aminoglycosides may be associated with aminoglycoside-modifying enzyme gene expressions.

  16. Mechanistic studies of copper(II)-aminoglycoside mediated DNA damage and magnesium catalyzed nuclease activity of hammerhead ribozyme

    Science.gov (United States)

    Patwardhan, Anjali A.

    The antibacterial activity of aminoglycosides stems from their high affinity binding to the 16S rRNA in bacteria resulting in inhibition of protein synthesis. Used to treat acute bacterial infections these antibiotics have limited applications due to their high dosage requirements and the emergence of resistant strains. We have synthesized and characterized Cu(II) derivatives of the aminoglycosides, kanamycin A, tobramycin, neamine, kanamycin B, neomycin B, and paromomycin. The first three exhibit preferential and tight binding to Cu(II) as against neomycin B and kanamycin B and paromomycin. EPR of frozen solutions and UV-visible spectroscopy suggest a change in geometry around the Cu(II) but the stabilities of the complexes in water differ. These copper derivatives efficiently cleave plasmid DNA at micromolar concentrations (hydrolytic) and at nanomolar concentrations in the presence co-reactants like hydrogen peroxide or ascorbic acid. Hydrolysis is multi turnover and exhibits Michelis-Menten kinetics with enzyme-like behavior whereas oxidative cleavage is highly specific with C-4' H abstraction resulting in characteristic base propenal and nucleotide base products. Hydroxyl radicals generated are copper based and are generated in close proximity of the substrate. Hammerhead ribozymes are selectively hydrolyzed in the presence of divalent ions with Mg2+ being the metal ion of choice in vivo . Our studies with complex ions like cobalt hexaammine and fac-triamminetriaquochromium(III) establish outer sphere interactions of Mg2+ with the hammerhead in the catalytic site. There are two sets of sites, one structural and one catalytic. Complex ions in the catalytic site and divalent ions in the structural site result in a slow but active hammerhead ribozyme suggesting that the complex ions are not inhibitory, contrary to what was suggested previously.

  17. aph(3')-IIb, a gene encoding an aminoglycoside-modifying enzyme, is under the positive control of surrogate regulator HpaA.

    Science.gov (United States)

    Zeng, Lin; Jin, Shouguang

    2003-12-01

    Pseudomonas aeruginosa harbors a chromosomal aminoglycoside phosphotransferase gene, aph(3')-IIb, which confers P. aeruginosa resistance to several important aminoglycoside antibiotics, including kanamycin A and B, neomycin B and C, butirosin, and seldomycin F5. The aph(3')-IIb gene has been found to be regulated by an AraC-type transcriptional regulator (HpaA) encoded by a gene located upstream of the aph(3')-IIb gene. In the presence of 4-hydroxyphenylacetic acid (4-HPA), HpaA activates the expression of aph(3')-IIb as well as that of the hpa regulon which encodes metabolic enzymes for the utilization of 4-HPA. hpaA and aph(3')-IIb form an operon, and in response to the presence of 4-HPA, the wild-type P. aeruginosa strain PAK (but not its hpaA mutant strain) displays increased resistance to neomycin. A survey of 39 clinical and 19 environmental isolates of P. aeruginosa demonstrated in all of them the presence of an hpaA-aph gene cluster, while 56 out of the 58 isolates are able to utilize the 4-HPA as a sole carbon source, suggesting a feature common to P. aeruginosa strains. Interestingly, a larger portion of clinical isolates than environmental isolates showed 4-HPA-induced resistance to neomycin. The aph(3')-IIb gene product is likely to function as a metabolic enzyme which has a cross-reactivity with aminoglycosides. These findings provide new insight into the possible mechanism of P. aeruginosa antibiotic resistance.

  18. Transgenic superroots of Lotus corniculatus can be regenerated from superroot-derived leaves following Agrobacterium-mediated transformation.

    Science.gov (United States)

    Tanaka, Hidenori; Toyama, Jun; Hashiguchi, Masatsugu; Kutsuna, Yasuyo; Tsuruta, Shin-ichi; Akashi, Ryo; Hoffmann, Franz

    2008-08-25

    Super-growing roots (superroots; SR), which have been established in the legume species Lotus corniculatus, are a fast-growing root culture that allows continuous root cloning, direct somatic embryogenesis and mass regeneration of plants under entirely growth regulator-free culture conditions. These features are unique for non-hairy root cultures, and they are now stably expressed since the culture was isolated more than 10 years ago (1997). Attempts to achieve direct and stable transformation of SR turned out to be unsuccessful. Making use of the supple regeneration plasticity of SR, we are reporting here an indirect transformation protocol. Leaf explants, derived from plants regenerated from SR, were inoculated with Agrobacterium tumefaciens strain LBA4404 harboring the binary vector pBI121, which contains the neomycin phosphotransferase II (NPTII) and beta-glucuronidase (GUS) genes as selectable and visual markers, respectively. After co-cultivation, the explants were selected on solidified MS medium with 0.5 mg/L benzylamino purine (BAP), 100 mg/L kanamycin and 250 mg/L cefotaxime. Kanamycin-resistant calli were transferred to liquid rooting medium. The newly regenerated, kanamycin-resistant roots were harvested and SR cultures re-established, which exhibited all the characteristics of the original SR. Furthermore, kanamycin-resistant roots cultured onto solidified MS medium supplemented with 0.5 mg/L BAP produced plants at the same rate as control SR. Six months after gene transfer, PCR analysis and histochemical locating indicated that the NPTII gene was integrated into the genome and that the GUS gene was regularly expressed in leaves, roots and nodules, respectively. The protocol makes it now possible to produce transformed SR and nodules as well as transgenic plants from transformed SR.

  19. Urinary gamma-glutamyl transferase as an indicator of acute nephrotoxicity in rats

    Energy Technology Data Exchange (ETDEWEB)

    Dierickx, P.J.

    1981-06-01

    A series of nephrotoxic compounds dissolved in 0.9% NaCl was given to groups of five male Wistar rats in a single i.p. injection. Mercuric acetate and mercuric trifluoroacetate at 1 mg Hg/kg induced a sharp increase in urinary gamma-glutamyl transferase (GGT) activity on day 1, followed by a decrease below control values on day 3. Sodium ethylmercurithiosalicylate induced a relatively small urinary GGT increase, explained by its low Hg-bioavailability. An increased urinary GGT activity was noted after treatment with the aminoglycoside antibiotics kanamycin, neomycin, paramomycin, and streptomycin, ammonium fluoride, potassium bichromate, sodium tetrathionate, and cis-diamminedichloroplatinum. This was lower than for the mercury compounds, but clearly different from the controls. The urinary GGT increase was an acute phenomenon. It is concluded that the measurement of urinary GGT can be used as an indicator of acute nephrotoxicity.

  20. Antimicrobial Activity and Antibiotic Sensitivity of Three Isolates of Lactic Acid Bacteria From Fermented Fish Product, Budu

    Directory of Open Access Journals (Sweden)

    Liasi, S. A.

    2009-01-01

    Full Text Available Three isolates of lactic acid bacteria (LAB from the fermented food product, Budu, were identified as genus lactobacillus (Lactobacillus casei LA17, Lactobacillus plantarum LA22 and L. paracasei LA02, and the highest population was Lb. paracasei LA02. The antibacterial agent produced by the isolates inhibited the growth of a range of gram-positive and gram-negative microorganisms. Antimicrobial sensitivity test to 18 different types of antibiotic were evaluated using the disc diffusion method. Inhibition zone diameter was measured and calculated from the means of five determinations and expressed in terms of resistance or susceptibility. All the LAB isolates were resistant to colestin sulphate, streptomycin, amikacin, norfloxacin, nalidixic acid, mecillinam, sulphanethoxazole/ trimethoprim, kanamycin, neomycin, bacitracin and gentamycin but susceptible to erythromycin, penicillin G, chloramphenicol, tetracycline, ampicillin and nitrofurantion.

  1. Agrobacterium-mediated transformation and assessment of factors influencing transgene expression in loblolly pine (Pinus taeda L.)

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    This investigation reports a protocol for transfer and expression of foreign chimeric genes in loblolly pine (Pinus taeda L.). Transformation was achieved by co-cultivation of mature zygotic embryos with Agrobacterium tumefaciens strain LBA4404 which harbored a binary vector (pBI121) including genes for 3-glucuronidase (GUS) and neomycin phosphotransferase (NPTI1). Factors influencing transgene expression including seed sources of loblolly pine, concentration of bacteria, and the wounding procedures of target explants were investigated. The expression of foreign gene was confirmed by the ability of mature zygotic embryos to produce calli in the presence of kanamycin, by histochemical assays of GUS activity, by PCR analysis, and by Southern blot. The successful expression of the GUS gene in different families of loblolly pine suggests that this transformation system is probably useful for the production of the genetically modified conifers.

  2. A random sequential mechanism of aminoglycoside acetylation by Mycobacterium tuberculosis Eis protein.

    Directory of Open Access Journals (Sweden)

    Oleg V Tsodikov

    Full Text Available An important cause of bacterial resistance to aminoglycoside antibiotics is the enzymatic acetylation of their amino groups by acetyltransferases, which abolishes their binding to and inhibition of the bacterial ribosome. Enhanced intracellular survival (Eis protein from Mycobacterium tuberculosis (Mt is one of such acetyltransferases, whose upregulation was recently established as a cause of resistance to aminoglycosides in clinical cases of drug-resistant tuberculosis. The mechanism of aminoglycoside acetylation by MtEis is not completely understood. A systematic analysis of steady-state kinetics of acetylation of kanamycin A and neomycin B by Eis as a function of concentrations of these aminoglycosides and the acetyl donor, acetyl coenzyme A, reveals that MtEis employs a random-sequential bisubstrate mechanism of acetylation and yields the values of the kinetic parameters of this mechanism. The implications of these mechanistic properties for the design of inhibitors of Eis and other aminoglycoside acetyltransferases are discussed.

  3. Properties of Achromobacter xylosoxidans highly resistant to aminoglycoside antibiotics.

    Science.gov (United States)

    Nakamoto, Sachiko; Goda, Natsumi; Hayabuchi, Tatsuya; Tamaki, Hiroo; Ishida, Ayami; Suzuki, Ayaka; Nakano, Kaori; Yui, Shoko; Katsumata, Yuto; Yamagami, Yuki; Burioka, Naoto; Chikumi, Hiroki; Shimizu, Eiji

    2016-04-01

    We herein discovered a highly resistant clinical isolate of Pseudomonas aeruginosa with MICs to amikacin, gentamicin, and arbekacin of 128 μg/mL or higher in a drug sensitivity survey of 92 strains isolated from the specimens of Yoka hospital patients between January 2009 and October 2010, and Achromobacter xylosoxidans was separated from this P. aeruginosa isolate. The sensitivity of this bacterium to 29 antibiotics was investigated. The MICs of this A. xylosoxidans strain to 9 aminoglycoside antibiotics were: amikacin, gentamicin, arbekacin, streptomycin, kanamycin, neomycin, and spectinomycin, 1,024 μg/mL or ≥ 1,024 μg/mL; netilmicin, 512 μg/mL; and tobramycin, 256 μg/mL. This strain was also resistant to dibekacin. This aminoglycoside antibiotic resistant phenotype is very rare, and we are the first report the emergence of A. xylosoxidans with this characteristic.

  4. Validated spectrofluorimetric method for determination of selected aminoglycosides

    Science.gov (United States)

    Omar, Mahmoud A.; Ahmed, Hytham M.; Hammad, Mohamed A.; Derayea, Sayed M.

    2015-01-01

    New, sensitive, and selective spectrofluorimetric method was developed for determination of three aminoglycoside drugs in different dosage forms, namely; neomycin sulfate (NEO), tobramycin (TOB) and kanamycin sulfate (KAN). The method is based on Hantzsch condensation reaction between the primary amino group of aminoglycosides with acetylacetone and formaldehyde in pH 2.7 yielding highly yellow fluorescent derivatives measured emission (471 nm) and excitation (410 nm) wavelengths. The fluorescence intensity was directly proportional to the concentration over the range 10-60, 40-100 and 5-50 ng/mL for NEO, TOB and KAN respectively. The proposed method was applied successfully for determination of these drugs in their pharmaceutical dosage forms.

  5. Transfer DREB into Lolium perenne L. To improve its drought tolerance

    Institute of Scientific and Technical Information of China (English)

    Ma Xinrong; Sun Zhenyuan; Jiang Changshun; Dong Zhaoyong; Zhang Yizheng

    2006-01-01

    A method of Agrobacterium tumefaciens mediated transformation for perennial ryegrass was developed using the calli of ryegrass derived from mature embryos. The calli were inoculated with a disarmed A. tumefaciens strain EHA105 harboring binary vector p2328. Vector p2328 contained transcription factor DREB1B and neomycin phosphotransferase (npt II) genes which were driven by promoters of rd29B and CaMV35S, respectively. The inoculated calli were selected on paromomycin- or kanamycin-containing media till the established plants being transferred to soil. Six transgenic plants with DREB1B had been obtained from perennial ryegrass strain Tove. PCR and Southern-blotting showed that npt II and DREB1B genes were integrated in perennial ryegrass genome. Stress treatment confirmed that transgenic plants with higher drought tolerance were obtained.

  6. Molecular epidemiological survey on aminoglycoside antibiotics-resistant genotype and phenotype of avian Escherichia coli in North China.

    Science.gov (United States)

    Zhang, T; Wang, C G; Jiang, G E; Lv, J C; Zhong, X H

    2012-10-01

    Monitoring drug resistance in Escherichia coli is important for prevention and treatment of colibacillosis. To choose effective drugs to prevent and control avian colibacillosis in North China, we investigated resistance of 205 E. coli isolates (from Beijing, Tianjin, inner Mongolia, Shanxi, and Hebei regions) to commonly used clinical aminoglycoside antibiotics using a drug susceptibility test. The results show that the isolates had varying degrees of resistance to kanamycin, gentamicin, streptomycin, amikacin, neomycin, and spectinomycin. Particularly, the resistance rates of the former 3 antibiotics exceeded 40%. To explore the reasons for wide drug resistance, aminoglycosides modifying enzymes (AME) genes, which are important in generation of aminoglycoside resistance, were detected by PCR. Of the isolates, 60.98% carried AME genes and 38.05% carried commensal multidrug resistance genes. Therefore, resistance of avian E. coli to aminoglycoside antibiotics is very serious in North China, perhaps due to the existence of resistance genes.

  7. [Antibiotic resistance in strains of Salmonella typhimurium and S. enteritidis isolated from poultry in the Czech Republic 1991-1992].

    Science.gov (United States)

    Cízek, A; Kovarík, K

    1994-01-01

    Antibiotic resistance has been monitored in 293 strains of S. typhimurium and 260 strains of S. enteritidis isolated from poultry in Czech Republic in the years 1991 and 1992. Ninety per cent of all salmonella isolations examined by disc diffusion method (Bauer et al., 1966) were sensitive to all 8 antimicrobials (chloramphenicol, neomycin, tetracycline, streptomycin, colistin, ampicillin, kanamycin, sulfisoxazol) used for testing. The strains of S. typhimurium were more resistant than S. enteritidis strains, as seen from the percentage of resistant strains, 17.4% and 1.2% respectively. Thirty-two (62.7%) out of 51 resistant strains were multiresistant. The percentage of resistance in S. typhimurium strains was as follows: sulfisoxazol (12.3%), streptomycin (11.3%), tetracycline (4.4%) and chloramphenicol (1.7%).

  8. Pseudomonas fluorescens associated with Bacterial Disease in Catla catla in Marathwada Region of Maharashtra

    Directory of Open Access Journals (Sweden)

    1Omprakash Darak

    2015-05-01

    Full Text Available In the present study a detailed analysis was carried out to evaluate the association of various bacterial pathogens with Catla catla from Marathwada region of Maharashtra. The freshwater fishes were collected from different water bodies and fish culturing centre of eight districts of Marathwada region viz. Aurangabad, Jalna, Parbhani, Nanded, Hingoli, Latur, Beed and Osmanabad. The analysis could yield thirteen pathogenic bacteria from the fish samples that included Micrococcus sp., Bacillus sp., Lactobacillus sp., Vibrio sp., Aeromonas sp., Streptococcussp., Flavobacterium sp., Vibrio sp., Proteus sp., Staphylococcus sp., Enterobacteria sp., E.coli , Pseudomonas sp. The bacterial strains were identified based on colony morphology, cell morphology and biochemical chemical characters. The dominant bacterial pathogen was Pseudomonas sp. The Pseudomonas sp associated with Catla catla could survive on host as well as in water. Pseudomonas fluorescens was very sensitive to Kanamycin, Nalidixic acid, Gentamicin and Neomycin, less sensitive to tetracycline, amikacin and Chlorophenicol and very less sensitive to Oxytetracycline, Erythromycin and Penicillin.

  9. Agrobacterium tumefaciens-mediated transformation of embryogenic tissue and transgenic plant regeneration in Chamaecyparis obtusa Sieb. et Zucc.

    Science.gov (United States)

    Taniguchi, T; Kurita, M; Ohmiya, Y; Kondo, T

    2005-03-01

    A genetic transformation procedure for Chamaecyparis obtusa was developed after co-cultivation of embryogenic tissues with disarmed Agrobacterium tumefaciens strain C58/pMP90, which harbours the sgfp (synthetic green fluorescent protein) visual reporter and nptII (neomycin phoshotransferase II) selectable marker genes. The highest transformation frequency was 22.5 independent transformed lines per dish (250 mg embryogenic tissue) following selection on kanamycin medium. Transgenic plantlets were regenerated through the maturation and germination of somatic embryos. The intensity of GFP fluorescence, observed under a fluorescence microscope, varied from very faint to relatively strong, depending on the transgenic line or part of the transgenic plant. The integration of the genes into the genome of regenerated plantlets was confirmed by Southern blot analysis.

  10. Study on the Factors Influencing the Efficiency of Wheat Transformation

    Institute of Scientific and Technical Information of China (English)

    YE Xing-guo; XU Hui-jun; DU Li-pu; XIN Zhi-yong

    2002-01-01

    Wheat transformation efficiency is closely related to several factors such as receptor genotype, constructed plasmid and selection procedure after bombardment or co-cultivation. In our study, several kinds of antibiotics, which were normally used in plant transformation to the selection genes of nptⅡ, bar and hpt,were tested for the optimal concentrations for wheat transformation. The results showed that 25 - 50mg/L of geneticin (G418) was suitable for the selection of nptⅡ, kanamycin or neomycin was not suitable for use. 3 5mg/L of phosphinothricin (PPT) or biolaphos could be used for the selection of bar, 100 - 150mg/L of hygromycin for the selection of hpt. Yangmai 158 and Yangmai 10 with high tissue culture response and good agronomic characteristics were screened from 25 potential Chinese wheat cultivars. The concentration changing of selectable agent in selection medium was helpful to obtain enough regeneration plantlets with strong root system.

  11. Antibiotic resistance of lactic acid bacteria isolated from Chinese yogurts.

    Science.gov (United States)

    Zhou, N; Zhang, J X; Fan, M T; Wang, J; Guo, G; Wei, X Y

    2012-09-01

    The aim of this study was to evaluate the susceptibility of 43 strains of lactic acid bacteria, isolated from Chinese yogurts made in different geographical areas, to 11 antibiotics (ampicillin, penicillin G, roxithromycin, chloramphenicol, tetracycline, chlortetracycline, lincomycin, kanamycin, streptomycin, neomycin, and gentamycin). The 43 isolates (18 Lactobacillus bulgaricus and 25 Streptococcus thermophilus) were identified at species level and were typed by random amplified polymorphic DNA analysis. Thirty-five genotypically different strains were detected and their antimicrobial resistance to 11 antibiotics was determined using the agar dilution method. Widespread resistance to ampicillin, chloramphenicol, chlortetracycline, tetracyclines, lincomycin, streptomycin, neomycin, and gentamycin was found among the 35 strains tested. All of the Strep. thermophilus strains tested were susceptible to penicillin G and roxithromycin, whereas 23.5 and 64.7% of Lb. bulgaricus strains, respectively, were resistant. All of the Strep. thermophilus and Lb. bulgaricus strains were found to be resistant to kanamycin. The presence of the corresponding resistance genes in the resistant isolates was investigated through PCR, with the following genes detected: tet(M) in 1 Lb. bulgaricus and 2 Strep. thermophilus isolates, ant(6) in 2 Lb. bulgaricus and 2 Strep. thermophilus isolates, and aph(3')-IIIa in 5 Lb. bulgaricus and 2 Strep. thermophilus isolates. The main threat associated with these bacteria is that they may transfer resistance genes to pathogenic bacteria, which has been a major cause of concern to human and animal health. To our knowledge, the aph(3')-IIIa and ant(6) genes were found in Lb. bulgaricus and Strep. thermophilus for the first time. Further investigations are required to analyze whether the genes identified in Lb. bulgaricus and Strep. thermophilus isolates might be horizontally transferred to other species.

  12. Aminoglycoside resistance rates, phenotypes, and mechanisms of Gram-negative bacteria from infected patients in upper Egypt.

    Science.gov (United States)

    Gad, Gamal F; Mohamed, Heba A; Ashour, Hossam M

    2011-02-17

    With the re-emergence of older antibiotics as valuable choices for treatment of serious infections, we studied the aminoglycoside resistance of Gram-negative bacteria isolated from patients with ear, urinary tract, skin, and gastrointestinal tract infections at Minia university hospital in Egypt. Escherichia coli (mainly from urinary tract and gastrointestinal tract infections) was the most prevalent isolate (28.57%), followed by Pseudomonas aeruginosa (25.7%) (mainly from ear discharge and skin infections). Isolates exhibited maximal resistance against streptomycin (83.4%), and minimal resistance against amikacin (17.7%) and intermediate degrees of resistance against neomycin, kanamycin, gentamicin, and tobramycin. Resistance to older aminoglycosides was higher than newer aminoglycosides. The most common aminoglycoside resistance phenotype was that of streptomycin resistance, present as a single phenotype or in combination, followed by kanamycin-neomycin as determined by interpretative reading. The resistant Pseudomonas aeruginosa strains were capable of producing aminoglycoside-modifying enzymes and using efflux as mechanisms of resistance. Using checkerboard titration method, the most frequently-observed outcome in combinations of aminoglycosides with β-lactams or quinolones was synergism. The most effective combination was amikacin with ciprofloxacin (100% Synergism), whereas the least effective combination was gentamicin with amoxicillin (53.3% Synergistic, 26.7% additive, and 20% indifferent FIC indices). Whereas the studied combinations were additive and indifferent against few of the tested strains, antagonism was never observed. The high resistance rates to aminoglycosides exhibited by Gram-negative bacteria in this study could be attributed to the selective pressure of aminoglycoside usage which could be controlled by successful implementation of infection control measures.

  13. Aminoglycoside resistance rates, phenotypes, and mechanisms of Gram-negative bacteria from infected patients in upper Egypt.

    Directory of Open Access Journals (Sweden)

    Gamal F Gad

    Full Text Available With the re-emergence of older antibiotics as valuable choices for treatment of serious infections, we studied the aminoglycoside resistance of Gram-negative bacteria isolated from patients with ear, urinary tract, skin, and gastrointestinal tract infections at Minia university hospital in Egypt. Escherichia coli (mainly from urinary tract and gastrointestinal tract infections was the most prevalent isolate (28.57%, followed by Pseudomonas aeruginosa (25.7% (mainly from ear discharge and skin infections. Isolates exhibited maximal resistance against streptomycin (83.4%, and minimal resistance against amikacin (17.7% and intermediate degrees of resistance against neomycin, kanamycin, gentamicin, and tobramycin. Resistance to older aminoglycosides was higher than newer aminoglycosides. The most common aminoglycoside resistance phenotype was that of streptomycin resistance, present as a single phenotype or in combination, followed by kanamycin-neomycin as determined by interpretative reading. The resistant Pseudomonas aeruginosa strains were capable of producing aminoglycoside-modifying enzymes and using efflux as mechanisms of resistance. Using checkerboard titration method, the most frequently-observed outcome in combinations of aminoglycosides with β-lactams or quinolones was synergism. The most effective combination was amikacin with ciprofloxacin (100% Synergism, whereas the least effective combination was gentamicin with amoxicillin (53.3% Synergistic, 26.7% additive, and 20% indifferent FIC indices. Whereas the studied combinations were additive and indifferent against few of the tested strains, antagonism was never observed. The high resistance rates to aminoglycosides exhibited by Gram-negative bacteria in this study could be attributed to the selective pressure of aminoglycoside usage which could be controlled by successful implementation of infection control measures.

  14. Drug resistance, plasmids, biotypes and susceptibility to bacteriophages of Salmonella isolated from poultry in Canada.

    Science.gov (United States)

    Poppe, C; McFadden, K A; Demczuk, W H

    1996-07-01

    Salmonella isolates from 295 layer and 294 broiler flocks in Canada were examined to determine resistance to antimicrobial agents, plasmid profiles, biochemical properties, and susceptibility to polyvalent bacteriophages. Except for the high number of strains resistant to spectinomycin (97.8%), the frequency of drug resistance of Salmonella isolates from layer flocks was low. None of 457 isolates from layer flocks was resistant to amikacin or ciprofloxacin, and less than 2% of the strains were resistant to cephalothin, chloramphenicol, cotrimoxazole, gentamicin, kanamycin, neomycin, nitrofurantoin, and/or polymyxin B. About 3% of the strains were resistant to ampicillin, carbenicillin and/or tetracycline, whereas 8% of the strains were resistant to sulfisoxazole. Salmonella anatum var. O15+ and S. typhimurium var. copenhagen strains were resistant to multiple antimicrobial agents. None of 1159 Salmonella strains from broiler flocks was resistant to amikacin, cephalothin, ciprofloxacin or polymyxin B, less than 1% of the strains were resistant to chloramphenicol, 2% were resistant to ampicillin, carbenicillin and/or chloramphenicol; 5-7% were resistant to the aminoglycosides gentamicin, kanamycin and/or neomycin; 6% were resistant to nitrofurantoin; 10% to tetracycline; 14% to sulfisoxazole; and 99% to spectinomycin. A high percentage of S. binza, S. anatum var. O15+, S. schwarzengrund and S. heidelberg strains were resistant to antimicrobial agents. Some of the single or multiple resistances were encoded by conjugative plasmids or by plasmids that were thermosensitive for transfer. Eight percent of S. heidelberg strains did not produce hydrogen sulfide. Ninety-seven percent of the Salmonella strains were susceptible to the lytic effect of polyvalent bacteriophages.

  15. Determination of neomycin in neomycin sulfate eye drops by rapid colorimetric assay%快速比色法测定硫酸新霉素滴眼剂的含量

    Institute of Scientific and Technical Information of China (English)

    何梅凤; 吴伟; 王延东; 唐细兰

    2006-01-01

    目的建立快速测定硫酸新霉素滴眼剂含量的方法.方法根据新霉素在碱性条件下能与Cu2+形成蓝色复合物,并有稳定的光吸收的原理,采用比色法测定硫酸新霉素滴眼剂含量,检测波长为594nm.结果新霉素在0.10万u~0.60万u/ml浓度范围内吸收度与浓度呈良好的线性关系,相关系数r=0.9998,平均回收率为100.67%,RSD为0.74%.结论本方法简单、快速、准确、重现性好,适合于产品中间体的质量控制和医院制剂的快速分析.

  16. Determination of neomycin in compound neomycin ear drops by HPLC with pre-column derivatization%HPLC法柱前衍生法测定复方新霉素滴耳液中新霉素的含量

    Institute of Scientific and Technical Information of China (English)

    陈蓉; 付丽娟

    2004-01-01

    目的:测定复方新霉素滴耳液中新霉素的含量.方法:用2,4-二硝基氟苯作柱前衍生剂,乙腈-三乙胺溶液(pH3.6)(60:40)为流动相,检测波长为365nm.结果:测得线性范围2.5~167μg·ml-1(r=0.9999),平均回收率为100.3%,RSD=0.82%(n=6).结论:本法操作简单,快速,灵敏度高.可分离新霉素B、C.

  17. 新霉素氟轻松乳膏中新霉素效价测定方法探讨%Determination Method of Potency of Neomycin in Neomycin Sulfate-fluocinolone Cream

    Institute of Scientific and Technical Information of China (English)

    卢翔; 程樱

    2007-01-01

    目的:研究新霉素氟轻松乳膏中硫酸新霉素的效价测定方法,扩大药品的检测范围,提高检测灵敏度.方法:扩大抗生素检测的浓度范围,以抗生素高低剂量浓度为6~3 u·ml-1或5~2.5 u·ml-1对新霉素氟轻松乳膏中新霉素效价进行测定.结果:与《中国药典》方法比较,无显著差异,但平均可信限率良好.结论:改进方法简便精密适用于新霉素氟轻松乳膏的效价测定.

  18. Determination of Neomycin in Triamcinolone Acetonide and Neomycin Paste by HPLC-ELSD%HPLC-ELSD法测定曲安奈德新霉素贴膏中新霉素含量

    Institute of Scientific and Technical Information of China (English)

    张玉婵; 廖志强; 郭环娟

    2015-01-01

    目的 建立采用HPLC-蒸发光散射检测器(ELSD)法测定曲安奈德新霉素贴膏中新霉素含量.方法 色谱条件为:Ultimate LP-C18色谱柱(250 mm×4.6 mm,5μm),流动相:0.2 mol/L三氟醋酸溶液-甲醇(95∶5),流速:1.0 mL/min,漂移管温度:40℃,载气流速:3.0 L/min,蒸发光检测器Gain值:4.结果 新霉素进样量在1.0~5.0 μg范围内,峰面积的对数值与进样量的对数值呈良好的线性关系(r=0.998 5).结论 方法准确、简便、重复性好,可用于曲安奈德新霉素贴膏中新霉素的含量测定.

  19. 新麻滴鼻液中盐酸麻黄碱与硫酸新霉素的含量测定%Determination of ephedrine hydrochloride and neomycin sulfate in the neomycin and ephedrine rasal drops

    Institute of Scientific and Technical Information of China (English)

    张翠莲; 张凡; 李大魁

    2005-01-01

    目的:建立新麻滴鼻液中盐酸麻黄碱和硫酸新霉素的含量测定方法.方法:用高效液相色谱法测定样品中盐酸麻黄碱的含量;用分光光度法测定样品中硫酸新霉素的含量.结果:盐酸麻黄碱在0.05~1.50 g·L-1范围内线性关系良好,r=0.999 8,平均回收率为101.28%(RSD≤1.16%);硫酸新霉素在60~420 IU·mL-1范围内线性关系较好,r=0.999 0,平均回收率为102.19%(RSD≤2.74%).结论:所建立的测定方法准确、重复性好、实用.

  20. Effective antibiotics against 'Candidatus Liberibacter asiaticus' in HLB-affected citrus plants identified via the graft-based evaluation.

    Science.gov (United States)

    Zhang, Muqing; Guo, Ying; Powell, Charles A; Doud, Melissa S; Yang, Chuanyu; Duan, Yongping

    2014-01-01

    Citrus huanglongbing (HLB), caused by three species of fastidious, phloem-limited 'Candidatus Liberibacter', is one of the most destructive diseases of citrus worldwide. To date, there is no established cure for this century-old and yet, newly emerging disease. As a potential control strategy for citrus HLB, 31 antibiotics were screened for effectiveness and phytotoxicity using the optimized graft-based screening system with 'Candidatus Liberibacter asiaticus' (Las)-infected citrus scions. Actidione and Oxytetracycline were the most phytotoxic to citrus with less than 10% of scions surviving and growing; therefore, this data was not used in additional analyses. Results of principal component (PCA) and hierarchical clustering analyses (HCA) demonstrated that 29 antibiotics were clustered into 3 groups: highly effective, partly effective, and not effective. In spite of different modes of actions, a number of antibiotics such as, Ampicillin, Carbenicillin, Penicillin, Cefalexin, Rifampicin and Sulfadimethoxine were all highly effective in eliminating or suppressing Candidatus Liberibacter asiaticus indicated by both the lowest Las infection rate and titers of the treated scions and inoculated rootstock. The non-effective group, including 11 antibiotics alone with three controls, such as Amikacin, Cinoxacin, Gentamicin, Kasugamycin, Lincomycin, Neomycin, Polymixin B and Tobramycin, did not eliminate or suppress Las in the tested concentrations, resulting in plants with increased titers of Las. The other 12 antibiotics partly eliminated or suppressed Las in the treated and graft-inoculated plants. The effective and non-phytotoxic antibiotics could be potential candidates for control of citrus HLB, either for the rescue of infected citrus germplasm or for restricted field application.

  1. Effective antibiotics against 'Candidatus Liberibacter asiaticus' in HLB-affected citrus plants identified via the graft-based evaluation.

    Directory of Open Access Journals (Sweden)

    Muqing Zhang

    Full Text Available Citrus huanglongbing (HLB, caused by three species of fastidious, phloem-limited 'Candidatus Liberibacter', is one of the most destructive diseases of citrus worldwide. To date, there is no established cure for this century-old and yet, newly emerging disease. As a potential control strategy for citrus HLB, 31 antibiotics were screened for effectiveness and phytotoxicity using the optimized graft-based screening system with 'Candidatus Liberibacter asiaticus' (Las-infected citrus scions. Actidione and Oxytetracycline were the most phytotoxic to citrus with less than 10% of scions surviving and growing; therefore, this data was not used in additional analyses. Results of principal component (PCA and hierarchical clustering analyses (HCA demonstrated that 29 antibiotics were clustered into 3 groups: highly effective, partly effective, and not effective. In spite of different modes of actions, a number of antibiotics such as, Ampicillin, Carbenicillin, Penicillin, Cefalexin, Rifampicin and Sulfadimethoxine were all highly effective in eliminating or suppressing Candidatus Liberibacter asiaticus indicated by both the lowest Las infection rate and titers of the treated scions and inoculated rootstock. The non-effective group, including 11 antibiotics alone with three controls, such as Amikacin, Cinoxacin, Gentamicin, Kasugamycin, Lincomycin, Neomycin, Polymixin B and Tobramycin, did not eliminate or suppress Las in the tested concentrations, resulting in plants with increased titers of Las. The other 12 antibiotics partly eliminated or suppressed Las in the treated and graft-inoculated plants. The effective and non-phytotoxic antibiotics could be potential candidates for control of citrus HLB, either for the rescue of infected citrus germplasm or for restricted field application.

  2. Assay of Neomycin Sulfate by polarimetry%旋光法测定硫酸新霉素含量

    Institute of Scientific and Technical Information of China (English)

    卢芳; 刘群; 余祥华; 金秀娥

    2006-01-01

    采用旋光法测定了硫酸新霉素含量.结果表明,在1 500~15 000 单位/mL的浓度范围内,旋光度与硫酸新霉素浓度呈良好线性关系,相关系数r=0.999 8(n=6).与微生物法相比,该方法具有简便、快速、易操作等优点.

  3. Testing the possibility of horizontal transfer of introduced neomycin phosphotransferase (nptⅡ) gene of transgenic Eucalyptus camaldulensis into soil bacteria

    Institute of Scientific and Technical Information of China (English)

    Katsuaki ISHII; LU Meng-zhu

    2008-01-01

    The possible horizontal transfer of transgenes is of great concern when the transgenic plants are released into the field. To test the possible transfer of nptⅡ of transgenic trees into soil bacteria, we have used a stool DNA preparation kit to isolate the DNA from the soils in the rhizospheres of two non- and eight transgenic Eucalyptus camaldulensis trees. All the samples have provided the corresponding PCR products in the amplification with bacterial 16S RNA specific sequences, which indicates that the quality of the isolated DNA is adequate for amplification. The nptⅡ specific band has been amplified in three soil samples from the transgenic trees and even treated with filtration before the DNA isolation. This indicates that nptlI DNA exists in the soil, although it is still unclear whether the DNA was in the soil particles, in the soil bacteria or in the Agrobaeterium contamination which was used for the E.camaldulensis transformation. Two approaches on isolation of bacterial DNA have been suggested for testing the possibility of this event in the future.

  4. 21 CFR 524.155 - Bacitracin zinc-polymyxin B sulfate-neomycin sulfate-hydrocortisone or hydrocortisone acetate...

    Science.gov (United States)

    2010-04-01

    ... thin film over the cornea three or four times daily. (2) Indications for use. For treating acute or... initial treatment of corneal ulcers. They should not be used until the infection is under control...

  5. Studies on the Conformational Features of Neomycin-B and its Molecular Recognition by RNA and Bacterial Defense Proteins

    Science.gov (United States)

    Asensio, Juan Luis; Bastida, Agatha; Jiménez-Barbero, Jesús

    According to NMR and molecular dynamics simulations, the conformational behavior of natural aminoglycosides is characterized by a remarkable flexibility, with different conformations, even non-exo-anomeric ones, in fast exchange. Very probably, this feature allows the adaptation of these ligands to the spatial and electronic requirements of different receptors. The large diversity of structures adopted by aminoglycosides in the binding pocket of the different RNA receptors and the distinct enzymes involved in bacterial resistance are consistent with this view. This conformational diversity can, in certain favorable cases, be exploited in the design of new antibiotic derivatives not susceptible to enzymatic inactivation, by designing tailor-made conformationally locked aminoglycosides.

  6. Toxicity modulation, resistance enzyme evasion, and A-site X-ray structure of broad-spectrum antibacterial neomycin analogs.

    Science.gov (United States)

    Maianti, Juan Pablo; Kanazawa, Hiroki; Dozzo, Paola; Matias, Rowena D; Feeney, Lee Ann; Armstrong, Eliana S; Hildebrandt, Darin J; Kane, Timothy R; Gliedt, Micah J; Goldblum, Adam A; Linsell, Martin S; Aggen, James B; Kondo, Jiro; Hanessian, Stephen

    2014-09-19

    Aminoglycoside antibiotics are pseudosaccharides decorated with ammonium groups that are critical for their potent broad-spectrum antibacterial activity. Despite over three decades of speculation whether or not modulation of pKa is a viable strategy to curtail aminoglycoside kidney toxicity, there is a lack of methods to systematically probe amine-RNA interactions and resultant cytotoxicity trends. This study reports the first series of potent aminoglycoside antibiotics harboring fluorinated N1-hydroxyaminobutyryl acyl (HABA) appendages for which fluorine-RNA contacts are revealed through an X-ray cocrystal structure within the RNA A-site. Cytotoxicity in kidney-derived cells was significantly reduced for the derivative featuring our novel β,β-difluoro-HABA group, which masks one net charge by lowering the pKa without compromising antibacterial potency. This novel side-chain assists in evasion of aminoglycoside-modifying enzymes, and it can be easily transferred to impart these properties onto any number of novel analogs.

  7. NPTII标记转基因棉花再生株的延其筛选

    Institute of Scientific and Technical Information of China (English)

    张慧军; 石跃进; 朱永红; 岳建雄; 杨怀义

    2002-01-01

    @@ 新霉素磷酸转移酶(Neomycin Phosphotransferase Ⅱ,NPTⅡ)基因是迄今植物遗传转化中应用最为广泛的选择标记.该基因编码新霉素磷酸转移酶亦称氨基糖苷-3'-磷酸转移酶(aminoglycoside-3'-phosphotransferase Ⅱ),其编码产物可使氨基糖苷类抗生素(aminoglycoside angibi-otics)如卡那霉素(kanamycin,Km)、新霉素(neomycin)、G418等磷酸化而失活.卡那霉素等氨基糖苷类抗生素能与植物细胞叶绿体和线粒体中的核糖体30S小亚基相结合,影响70S起始复合物的形成,干扰叶绿体及线粒体的蛋白质合成,从而导致植物细胞死亡.

  8. A new poultry semen extender. 4. Effect of antibacterials in control of bacterial contamination in chicken semen.

    Science.gov (United States)

    Sexton, T J; Jacobs, L A; McDaniel, G R

    1980-02-01

    Forty antibacterials were qualitatively and quantitatively tested for controlling aerobic bacterial contamination without affecting viability of semen. Semen samples were collected aseptically, diluted 1:4 with the Beltsville poultry semen extender containing one of 40 antibiotics and held for 0, 24, 48, and 72 hr at 5 C. Semen samples were monitored at each storage interval for bacterial counts, sperm motility, sperm counts, and fertilizing capacity. Gentamicin (2.5 microgram/ml), kanamycin (31.2 microgram/ml), neomycin (62.5 microgram/ml), and tobramycin (2.5 microgram/ml) were the only antibacterials tested which controlled microbial growth without affecting sperm viability for up to 24 hr storage at 5 C. Tobramycin maintained fertility equal to that of the non-antibiotic control up to 24 hr storage. Neomycin maintained higher levels of fertility in semen stored for 48 and 72 hr than in semen of controls to all other treatments for the same storage periods and to undiluted, unstored semen. The conclusion from these studies is that the control of aerobic microbial growth in chicken semen has little influence on the maintenance of sperm viability during low temperature storage.

  9. Molecular determinants of affinity for aminoglycoside binding to the aminoglycoside nucleotidyltransferase(2'')-Ia.

    Science.gov (United States)

    Wright, Edward; Serpersu, Engin H

    2006-08-29

    One of the most commonly occurring aminoglycoside resistance enzymes is aminoglycoside 2''-O-nucleotidyltransferase [ANT(2'')]. In the present study molecular determinants of affinity and specificity for aminoglycoside binding to this enzyme are investigated using isothermal titration calorimetry (ITC). Binding of aminoglycosides is enthalpically driven accompanied by negative entropy changes. The presence of metal-nucleotide increases the affinity for all but one of the aminoglycosides studied but has no effect on specificity. The substituents at positions 1, 2', and 6' are important determinants of substrate specificity. An amino group at these positions leads to greater affinity. No correlation is observed between the change in affinity and enthalpy. At the 2' position greater affinity results from a more negative enthalpy for an aminoglycoside containing an amino rather than a hydroxyl at that position. At the 6' position the greater affinity for an aminoglycoside containing an amino substituent results from a less disfavorable entropic contribution. The thermodynamic basis for the change in affinity at position 1 could not be determined because of the weak binding of one of the aminoglycoside substrates, amikacin. The effect of increasing osmotic stress on affinity was used to determine that a net release of approximately four water molecules occurs when tobramycin binds to ANT(2''). No measurable net change in the number of bound water molecules is observed when neomycin binds the enzyme. Data acquired in this work provide the rationale for the ability of ANT(2'') to confer resistance against kanamycins but not neomycins.

  10. Herbal therapy associated with antibiotic therapy: potentiation of the antibiotic activity against methicillin – resistant Staphylococcus aureus by Turnera ulmifolia L

    Directory of Open Access Journals (Sweden)

    Lima Edeltrudes O

    2009-05-01

    Full Text Available Abstract Background Staphylococcus genus is widely spread in nature being part of the indigenous microbiota of skin and mucosa of animal and birds. Some Staphylococcus species are frequently recognized as etiological agents of many animal and human opportunistic infections This is the first report testing the antibiotic resistance-modifying activity of Turnera ulmifolia against methicillin-resistant Staphylococcus aureus – MRSA strain. Methods In this study an ethanol extract of Turnera ulmifolia L. and chlorpromazine were tested for their antimicrobial activity alone or in combination with aminoglycosides against an MRSA strain. Results The synergism of the ethanol extract and aminoglycosides were verified using microdillution method. A synergistic effect of this extract on gentamicin and kanamycin was demonstrated. Similarly, a potentiating effect of chlorpromazine on kanamycin, gentamicin and neomycin, indicating the involvement of an efflux system in the resistance to these aminoglycosides. Conclusion It is therefore suggested that extracts from Turnera ulmifolia could be used as a source of plant-derived natural products with resistance-modifying activity, constituting a new weapon against the problem of bacterial resistance to antibiotics demonstrated in MRSA strains.

  11. Optimization of Agrobacterium mediated genetic transformation of cotyledonary node explants of Vigna radiata.

    Science.gov (United States)

    Yadav, Sushil Kumar; Katikala, Sweety; Yellisetty, Varalaxmi; Kannepalle, Annapurna; Narayana, Jyothi Lakshmi; Maddi, Vanaja; Mandapaka, Maheswari; Shanker, Arun Kumar; Bandi, Venkateswarlu; Bharadwaja, Kirti Pulugurtha

    2012-12-01

    A reproducible and highly efficient protocol for genetic transformation mediated by Agrobacterium has been established for greengram (Vigna radiata L. Wilczek). Double cotyledonary node (DCN) explants were inoculated with Agrobacterium tumefaciens strain LBA 4404 harboring a binary vector pCAMBIA 2301 containing neomycin phosphotransferase (npt II) gene as selectable marker, β-glucuronidase (GUS) as a reporter (uidA) gene and annexin 1 bj gene. Important parameters like optical density of Agrobacterium culture, culture quantity, infection medium, infection and co-cultivation time and acetosyringone concentration were standardized to optimize the transformation frequency. Kanamycin at a concentration of 100 mg/l was used to select transformed cells. Transient and stable GUS expressions were studied in transformed explants and regenerated putative plants, respectively. Transformed shoot were produced on regeneration medium containing 100 mg/l kanamycin and 250 mg/l cefotaxime and rooted on ½ MS medium. Transient and constitutive GUS expression was observed in DCN explants and different tissues of T(0) and T(1) plants. Rooted T(0) and T(1) shoots confirming Polymerase Chain Reaction (PCR) positive for npt II and annexin 1bj genes were taken to maturity to collect the seeds. Integration of annexin gene into the greengram genome was confirmed by Southern blotting.

  12. Stability of antibiotics under growth conditions for thermophilic anaerobes

    Energy Technology Data Exchange (ETDEWEB)

    Peteranderl, R.; Shotts, E.B. Jr.; Wiegel, J. (Univ. of Georgia, Athens (United States))

    1990-06-01

    It was shown that the inhibitory effect of kanamycin and streptomycin in a growing culture of Clostridium thermohydrosulfuricum JW 102 is of limited duration. To screen a large number of antibiotics, their stability during incubation under the growth conditions of thermophilic clostridia was determined at 72 and 50C by using a 0.2% yeast extract-amended prereduced mineral medium with a pH of 7.3 or 5.0. Half-lives were determined in a modified MIC test with Escherichia coli, Staphylococcus aureus, and Bacillus megaterium as indicator strains. All compounds tested were similar at the two temperatures or more stable at 50 than at 72C. The half-life (t{sub 1/2}) at pH 7.3 and 72C ranged from 3.3 h (k = 7.26 day{sup {minus}1}, where k (degradation constant) = 1/t{sub 1/2}) for ampicillin to no detectable loss of activity for kanamycin, neomycin, and other antibiotics. Apparently some compounds became more potent during incubation. A change to pH 5.0 caused some compounds to become more labile to become more stable than at pH 7.3.

  13. Axenic culture of free-living conchocelis of Porphyra yezoensis and Porphyra haitanensis

    Science.gov (United States)

    Liu, Hui-Lian; Shuai, Li; Duan, De-Lin; Xu, Huai-Shu

    2002-03-01

    After discarding marine microorganisms from conchocelis of Porphyra yezoensis and Porphyra haitanensis, their axenic cultures were obtained through treatment with antibiotics. Antibiotic disc tests were carried out to determine the effectiveness of each antibiotic in eliminating contaminating microorganisms. Five of 12 antibiotics tested were selected and used to produce the axenic cultures in this study, which showed that 200 μg/mL streptomycin, 250 μg/mL penicillin, 252 μg/mL kanamycin, 30 μg/mL chloramphenicol were effective concentrations for eliminating microorganisms from conchocelis when antibiotics were added singly step by step; whereas simultaneous combination of 150 μg/mL streptomycin, 250 (or 350) μg/mL penicillin, 150 (or 250) μg/mL kanamycin, 70 μg/mL neomycin and 200 μg/mL chloramphenicol was also effective for producing the axenic cultures. However, it seemed that the treatments with antibiotics applied individually were more feasible than those will all antibiotics added at the same time. This may be due to the combined inhibiting effect of antibiotics on the growth and development of conchocelis.

  14. Antibacterial and Antioxidant Activities of Ursolic Acid and Derivatives

    Directory of Open Access Journals (Sweden)

    Patrícia G.G. do Nascimento

    2014-01-01

    Full Text Available Ursolic acid, an important bioactive compound, was isolated from ethanol extract of aerial parts of Sambucus australis. In order to develop bioactive ursolic acid derivatives, two semi-synthetic compounds were obtained through modification at C-3. The antibacterial activity of the ursolic acid and its derivatives was investigated. The microdilution method was used for determination of the minimal inhibitory concentration (MIC, against twelve bacterial strains. The influence of ursolic acid and its derivatives on the susceptibility of some bacterial pathogens to the aminoglycosides antibiotics neomycin, amikacin, kanamycin and gentamicin was evaluated. The most representative synergistic effect was observed by 3β-formyloxy-urs-12-en-28-oic acid at the concentration of 64 μg/mL in combination with kanamycin against Escherichia coli (27, a multidrug-resistant clinical isolate from sputum, with reduction of MIC value from 128 μg/mL to 8 μg/mL. Ursolic acid and its derivatives were examined for their radical scavenger activity using the DPPH assay, and showed significant activity.

  15. Scientific Opinion on a request from the European Commission for the assessment of the scientific elements put forward by Hungary to support the prohibition for the placing on the market of GM potato EH92-527-1 for cultivation purposes in Hungary

    Directory of Open Access Journals (Sweden)

    EFSA Panel on Genetically Modified Organisms (GMO

    2012-12-01

    Full Text Available Hungary notified to the European Commission its scientific arguments justifying the implementation of a national safeguard measure prohibiting the placing on the market of GM potato EH92-527-1 for cultivation purposes in Hungary, after which the European Commission asked the European Food Safety Authority (EFSA to assess the scientific information supporting the prohibition. Having considered the information package provided by Hungary and all relevant scientific publications, the EFSA Panel on Genetically Modified Organisms (GMO Panel concluded that (i no new data specific to the safety of the nptII gene have been provided; (ii the therapeutic relevance of kanamycin and neomycin was already addressed in the previous EFSA opinion on antibiotic resistance marker genes and kanamycin resistance in Mycobacterium tuberculosis results largely from chromosomal mutations and not from the transfer of aminoglycoside resistance genes such as nptII; (iii the knowledge gaps and uncertainties highlighted in the Hungarian document have already been considered in the previous EFSA opinion on antibiotic resistance marker genes, and no new information on the safety of nptII gene as present in the GM potato EH92-527-1 has been identified in the scientific literature that would cause the GMO Panel to change its previous conclusions. Therefore, the EFSA GMO Panel concludes that no grounds exist to date that would lead to reconsideration of its opinion on GM potato EH92-527-1.

  16. Antimicrobial Sensitivity of Avibacterium paragallinarum Isolates from Four Latin American Countries.

    Science.gov (United States)

    Luna-Galaz, G A; Morales-Erasto, V; Peñuelas-Rivas, C G; Blackall, P J; Soriano-Vargas, E

    2016-09-01

    The antimicrobial sensitivity of 11 reference strains and 66 Avibacterium paragallinarum isolates from four Latin American countries was investigated. All 11 reference strains were sensitive to amoxicillin-clavulanic acid, ampicillin, fosfomycin, gentamicin, kanamycin, neomycin, penicillin, tetracycline, and trimethoprim-sulfamethoxazole. The 11 reference strains were all resistant to lincomycin. All isolates (100%) from Mexico, Panama, and Peru were sensitive to amoxicillin-clavulanic acid, ampicillin, and fosfomycin. The Ecuadorian isolates showed some level of resistance to all 16 agents tested. The Ecuadorian isolates were significantly more sensitive to erythromycin, lincomycin, and streptomycin, and significantly more resistant to gentamicin, kanamycin, penicillin, and tetracycline, than the Mexican isolates. A total of 57.5% (38/66) of tested isolates were multi-drug resistant (MDR), with 16 MDR patterns detected in 88.4% (23/26) of the antimicrobial-resistant isolates from Ecuador, and 8 MDR patterns detected in 42.8% (15/35) of the antimicrobial-resistant isolates from Mexico. In conclusion, the variation in antimicrobial sensitivity patterns between isolates from Ecuador and Mexico emphasizes the importance of active, ongoing monitoring of A. paragallinarum isolates.

  17. Regeneration of Transgenic Soybean (Glycine max) Plants from Electroporated Protoplasts.

    Science.gov (United States)

    Dhir, S K; Dhir, S; Savka, M A; Belanger, F; Kriz, A L; Farrand, S K; Widholm, J M

    1992-05-01

    Transgenic soybean (Glycine max [L.] Merr.) plants were regenerated from calli derived from protoplasts electroporated with plasmid DNA-carrying genes for a selectable marker, neomycin phosphotransferase (NPTII), under the control of the cauliflower mosaic virus 35-Svedberg unit promoter, linked with a nonselectable mannityl opine synthesis marker. Following electroporation and culture, the protoplast-derived colonies were subjected to kanamycin selection (50 micrograms per milliliter) beginning on day 15 for 6 weeks. Approximately, 370 to 460 resistant colonies were recovered from 1 x 10(6) electroporated protoplasts, giving an absolute transformation frequency of 3.7 to 4.6 x 10(-4). More than 80% of the kanamycin-resistant colonies showed NPTII activity, and about 90% of these also synthesized opines. This indicates that the linked marker genes were co-introduced and co-expressed at a very high frequency. Plants were regenerated from the transformed cell lines. Southern blot analysis of the transformed callus and leaf DNA demonstrated the integration of both genes. Single-plant assays performed with different plant parts showed that both shoot and root tissues express NPTII activity and accumulate opines. Experiments with NPTII and mannityl opine synthesis marker genes on separate plasmids resulted in a co-expression rate of 66%. These results indicate that electroporation can be used to introduce both linked and unlinked genes into the soybean to produce transformed plants.

  18. Genetic transformation of Pinus taeda by particle bombardment

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    A protocol is presented for genetically engineering loblolly pine (Pinus taeda L.) using particle bombardment. This protocol enabled the routine transformation of loblolly pine plants that were previously difficult to transform. Mature zygotic embryos were used to be bombarded and to generate organogenic callus and transgenic regenerated plants. Plasmid pB48.215 DNA contained a synthetic Bacillus thuringiensis (B.t.) cryIAc coding sequence flanked by the double cauliflower mosaic virus (CaMV) 35S promoter and nopaline synthase (Nos) terminator sequences, and the selectable marker gene, neomycin phosphotransferase II (nptII) controlled by the promoter of the nopaline synthase gene was introduced into loblolly pine tissues by particle bombardment. The transformed tissues were proliferated and selected by kanamycin resistance conferred by the introduced NPTII gene. Shoot regeneration was induced from the kanamycin-resistant callus, and transgenic plantlets were then produced. The presence of the introduced genes in the transgenic loblolly pine plants was confirmed by polymerase chain reactions (PCR) analysis, by Southern blot analysis, and insect feeding assays. The recovered transgenic plants were acclimatized and then established in soil.

  19. Antibiotic Resistance of Escherichia Coli Isolated From Poultry and Poultry Environment of Bangladesh

    Directory of Open Access Journals (Sweden)

    Muhammad A. Akond

    2009-01-01

    Full Text Available Problem statement: Increased emergence in microbial resistance to antibiotics is a growing problem in Bangladesh, a tropical country with a large agrarian population having limited medical facilities. Wide spread use of antimicrobials in poultry farming here is a concern of multi-drug microbial resistance development that can potentially be transmitted to human pathogens even from non-pathogenic carrier strains. Attempt was made to assess drug susceptibility in Escherichia coli from poultry sources of Bangladesh. Approach: Eighty selected strains isolated from poultry sources were thoroughly characterized by standard cultural and biochemical tests followed by final identification using latex agglutination test of polyvalent anti-sera, from which 50 were tested for susceptibility to 13 antibiotics following disk diffusion method. Results: 145 (58%, out of total 250, were found positive for E. coli. 52-88% of tested E. coli strains from poultry sources were found resistant to Penicillin, Ciprofloxacin, Riphampicin, Kanamycin, Streptomycin, Cefixine, Erythromycin, Ampicillin, Tetracycline, and 20% strains showed resistance to both Chloramphenicol and Neomycin. No strains showed resistance to Norfloxacin and Gentamicin. Sensitivity was recorded in case of 60-86% strains to Norfloxacin, Gentamicin, Chloramphenicol, and Neomycin; and 26-36% strains against Tetracycline, Streptomycin, and Ampicillin. Intermediate resistance/ susceptibility to various antibiotics were observed for 12-36% Escherichia coli strains. Both, resistance and susceptibility were exhibited against Chloramphenicol, Ampicillin, Gentamicin, Neomycin, Tetracycline, Streptomycin and Norfloxacin. Multi drug resistance was found in case of 6-10 antibiotics for all strains tested. Conclusion: Further study is required on the role of poultry borne bacteria as vectors in transmitting drug resistance. Attention is to be paid for personnel hygiene in processing and handling of poultry and

  20. aph(3′)-IIb, a Gene Encoding an Aminoglycoside-Modifying Enzyme, Is under the Positive Control of Surrogate Regulator HpaA

    Science.gov (United States)

    Zeng, Lin; Jin, Shouguang

    2003-01-01

    Pseudomonas aeruginosa harbors a chromosomal aminoglycoside phosphotransferase gene, aph(3′)-IIb, which confers P. aeruginosa resistance to several important aminoglycoside antibiotics, including kanamycin A and B, neomycin B and C, butirosin, and seldomycin F5. The aph(3′)-IIb gene has been found to be regulated by an AraC-type transcriptional regulator (HpaA) encoded by a gene located upstream of the aph(3′)-IIb gene. In the presence of 4-hydroxyphenylacetic acid (4-HPA), HpaA activates the expression of aph(3′)-IIb as well as that of the hpa regulon which encodes metabolic enzymes for the utilization of 4-HPA. hpaA and aph(3′)-IIb form an operon, and in response to the presence of 4-HPA, the wild-type P. aeruginosa strain PAK (but not its hpaA mutant strain) displays increased resistance to neomycin. A survey of 39 clinical and 19 environmental isolates of P. aeruginosa demonstrated in all of them the presence of an hpaA-aph gene cluster, while 56 out of the 58 isolates are able to utilize the 4-HPA as a sole carbon source, suggesting a feature common to P. aeruginosa strains. Interestingly, a larger portion of clinical isolates than environmental isolates showed 4-HPA-induced resistance to neomycin. The aph(3′)-IIb gene product is likely to function as a metabolic enzyme which has a cross-reactivity with aminoglycosides. These findings provide new insight into the possible mechanism of P. aeruginosa antibiotic resistance. PMID:14638496

  1. Drug: D08260 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available IVE AGENTS A07A INTESTINAL ANTIINFECTIVES A07AA Antibiotics A07AA01 Neomycin D08260 Neomycin (INN) B BLOOD A...5 Neomycin D08260 Neomycin (INN) R RESPIRATORY SYSTEM R02 THROAT PREPARATIONS R02A THROAT PREPARATIONS R02AB Antibiotics... R02AB01 Neomycin D08260 Neomycin (INN) S SENSORY ORGANS S01 OPHTHALMOLOGICALS S01A ANTIINFECTIVES S01AA Antibiotics

  2. Drug: D05141 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available SENSORY ORGANS S01 OPHTHALMOLOGICALS S01A ANTIINFECTIVES S01AA Antibiotics S01AA03 Neomycin D05141 Neomycin...J01GB05 Neomycin D05141 Neomycin palmitate (USAN) R RESPIRATORY SYSTEM R02 THROAT PREPARATIONS R02A THROAT PREPARATIONS R02AB Antibio...biotics A07AA01 Neomycin D05141 Neomycin palmitate (USAN...mycin palmitate (USAN) A07 ANTIDIARRHEALS, INTESTINAL ANTIINFLAMMATORY/ANTIINFECTIVE AGENTS A07A INTESTINAL ANTIINFECTIVES A07AA Anti...tics R02AB01 Neomycin D05141 Neomycin palmitate (USAN) S

  3. Relationship between antimicrobial resistance and aminoglycoside-modifying enzyme gene expressions in Acinetobacter baumannii

    Institute of Scientific and Technical Information of China (English)

    SHI Wei-feng; JIANG Jian-ping; MI Zu-huang

    2005-01-01

    Background Acinetobacter baumannii is one of the main gram-negative bacilli in clinical practice. Nosocomial infections caused by multi-drug resistance Acinetobacter baumannii is very difficult to treat. This study was designed to investigate the antimicrobial resistance characteristics and four resistant gene expressions of aminoglycoside-modifying enzymes including N-acetyltransferases and O-phosphotransferases in Acinetobacter baumannii. Methods Bacterial identification and antimicrobial susceptibility test were performed by PhoenixTM system in 247 strains of Acinetobacter baumannii. Minimal inhibitory concentrations (MICs) of seven aminoglycosides including gentamicin, amikacin, kanamycin, tobramycin, netilmicin, neomycin and streptomycin in 15 strains of multi-drug resistant Acinetobacter baumannii were detected by agar dilution. Four aminoglycoside-modifying enzyme genes were amplified by polymerase chain reaction (PCR) and verified by DNA sequencer.Results The resistance rates of 247 strains of Acinetobacter baumannii against cefotaxime, levofloxacin, piperacillin, aztreonam, tetracycline, ciprofloxacin and chloramphenicol were more than 50%. Imipenem and meropenem showed high antibacterial activities with resistance rates of 3.2% and 4.1%. MIC50 and MIC90 of gentamicin, amikacin, streptomycin and kanamycin in 15 strains of multi-drug resistant Acinetobacter baumanii were all more than 1024 mg/L, and the resistance rates were 100%, 100%, 100% and 93.3%, respectively. But their resistance rates to tobramycin, netilmicin and neomycin were 86.7%, 93.3% and 46.7%, respectively. Three modifying enzyme genes, including aacC1, aacC2 and aacA4 genes, were found in 15 strains, but aphA6 had not been detected. Their positive rates were 93.3%, 20.0% and 20.0%, respectively. These three genes existed simultaneously in No.19 strain. Nucleotide sequences of aacC1, aacC2 and aacA4 genes shared 100%, 97.9% and 99.7% identities with GenBank genes (AY307113, S68058 and AY

  4. Agrobacterium-Mediated Transformation of Glycine Max and Regene rati on of Transgenic Plants%根癌农杆菌介导β-1,4-半乳糖苷转移酶基因转化大豆及其转基因植株再生

    Institute of Scientific and Technical Information of China (English)

    张毅; 李弘剑; 张俊辉; 郭勇

    2001-01-01

    A reproducible transformation system was developed for soybean (Glycin e max) using as explants sections from the excised hypocotyls of seeds. A constr uct containing cauliflower mosaic virus 35S-neomycin phosphotransferase (nptII) and human β-1,4-galactosyltransferase (hGT) was introduced into soybean culti var using Agrobacterium tumefaciens- mediated transfermation procedures. Regenerat i on was via organogenesis and transformed plants were selected on medium containi ng 50 mg/L kanamycin and 100 mg/L of aminopenicillanic acia. Transgenic soybeans were raised in the glasshouse. The hGT genes was integrated into the chomosomal genome of primary transgenic soybean plants, The transformation of regenerated plants was confirmed by assays for neomycin phosphotransferase activity and Sout hern blotting analyses.%以大豆下胚轴为外植体,通过根癌农杆菌介导转化法,建立起良好的转化系统,将人的β-1,4-半乳糖苷转移酶基因(hGT)导入大豆。经转化的外植体在添加100mg/L氨苄青霉素和50mg/L卡那霉素的选择培养基中可诱导出愈伤组织和芽再生,在1/2MS培养基上诱导生根,并培养成再生苗,获得了完整的抗性再生植株。进行Southern blot分子杂交鉴定,证实了外源基因hGT已稳定地整合到植物基因组中。

  5. SEROVARS AND ANTIMICROBIAL RESISTANCE OF Salmonella spp. ISOLATED FROM TURKEY AND BROILER CARCASSES IN SOUTHERN BRAZIL BETWEEN 2004 AND 2006.

    Science.gov (United States)

    Palmeira, Andre; Santos, Luciana Ruschel dos; Borsoi, Anderlise; Rodrigues, Laura Beatriz; Calasans, Max; Nascimento, Vladimir Pinheiro do

    2016-01-01

    Salmonella spp. causes diseases in fowls, when species-specific serovars (Salmonella Pullorum and S.Gallinarum) are present in flocks, and public health problems, when non-typhoid serovars are isolated, as well as possible bacterial resistance induced by the preventive and therapeutic use of antimicrobials in animal production. This study describes the serovars and bacterial resistance of 280 Salmonella spp. strains isolated from turkey and broiler carcasses in Southern Brazil between 2004 and 2006. Salmonella Enteritidis was the most prevalent serovar (55.7%), followed by Heidelberg (5.0%), Agona (4.3%), Bredeney (3.9%), Hadar (3.2%), and Typhimurium (2.9%). Tennessee and S. Enterica subspecies enterica(O: 4.5) were isolated only in turkeys, and Hadar (18.6%) was the most prevalent serovar in this species. Antimicrobial susceptibility tests were performed in 178 isolates (43 from turkeys and 135 from broilers). All isolates were sensitive to amoxicillin + clavulanic acid, polymyxin B, ciprofloxacin, and norfloxacin, and were resistant to bacitracin and penicillin. Broiler carcass isolates showed resistance to nalidixic acid (48.9%), nitrofurantoin (34.3%), neomycin (9.6%), tetracycline (5.2%), and kanamycin (8.9%); and turkey carcass isolates were resistant to nalidixic acid (62.8%), tetracycline (34.9%), and neomycin (30.2%), with a significant difference in turkeys when compared to broiler carcass isolates. These results indicate the need for judicious use of antimicrobials in livestock production, given that the serovars identified are potential causes of food poisoning.

  6. SEROVARS AND ANTIMICROBIAL RESISTANCE OF Salmonella spp. ISOLATED FROM TURKEY AND BROILER CARCASSES IN SOUTHERN BRAZIL BETWEEN 2004 AND 2006

    Directory of Open Access Journals (Sweden)

    Andre PALMEIRA

    2016-01-01

    Full Text Available Salmonella spp. causes diseases in fowls, when species-specific serovars (Salmonella Pullorum and S.Gallinarum are present in flocks, and public health problems, when non-typhoid serovars are isolated, as well as possible bacterial resistance induced by the preventive and therapeutic use of antimicrobials in animal production. This study describes the serovars and bacterial resistance of 280Salmonella spp. strains isolated from turkey and broiler carcasses in Southern Brazil between 2004 and 2006. SalmonellaEnteritidis was the most prevalent serovar (55.7%, followed by Heidelberg (5.0%, Agona (4.3%, Bredeney (3.9%, Hadar (3.2%, and Typhimurium (2.9%. Tennessee and S. Enterica subspecies enterica(O: 4.5 were isolated only in turkeys, and Hadar (18.6% was the most prevalent serovar in this species. Antimicrobial susceptibility tests were performed in 178 isolates (43 from turkeys and 135 from broilers. All isolates were sensitive to amoxicillin + clavulanic acid, polymyxin B, ciprofloxacin, and norfloxacin, and were resistant to bacitracin and penicillin. Broiler carcass isolates showed resistance to nalidixic acid (48.9%, nitrofurantoin (34.3%, neomycin (9.6%, tetracycline (5.2%, and kanamycin (8.9%; and turkey carcass isolates were resistant to nalidixic acid (62.8%, tetracycline (34.9%, and neomycin (30.2%, with a significant difference in turkeys when compared to broiler carcass isolates. These results indicate the need for judicious use of antimicrobials in livestock production, given that the serovars identified are potential causes of food poisoning.

  7. Evidence on Possible Mycoplasma Etiology of Aster Yellows Disease II. Suppression of Aster Yellows in Insect Vectors.

    Science.gov (United States)

    Whitcomb, R F; Davis, R E

    1970-08-01

    Chlortetracycline or chloramphenicol (but not kanamycin, penicillin, or erythromycin), when administered in hydroponic solution to diseased aster, reduced the availability of the aster yellows (AY) agent to nymphs of Macrosteles fascifrons (Stål). Insects exposed to healthy plants whose roots were immersed in chlortetracycline were able to acquire AY agent from diseased plants the day after removal from the antibiotic-treated plants, but the latent period of the ensuing disease in the insects was prolonged. Chlortetracycline or tylosin tartrate blocked AY infection in nymphs injected with a mixture of antibiotic and the AY agent, but polymyxin, neomycin, vancomycin, penicillin, carbomycin, or chloramphenicol did not. All tetracyclines tested, methacycline, oxytetracycline, and chlortetracycline, produced a dramatic reduction in the ability of infected vectors to transmit AY agent. Tylosin tartrate also reduced transmission when injected into AY-transmitting vectors, but carbomycin, spectinomycin, cycloserine, penicillin, erythromycin, or kanamycin had no such effect. During the first 10 days after injection of tylosin tartrate or oxytetracycline into transmitting vectors, ability of the insects to transmit AY decayed rapidly. Transmission by insects injected with buffer alone, after decreasing the first day after injection, gradually returned to its normal level in less than 1 week. By 2 to 3 weeks after injection with tylosin or oxytetracycline, ability to transmit AY was regained by vectors. The results suggest that tetracycline antibiotics and tylosin tartrate inhibit multiplication of AY agent in the insect. The spectrum of antibiotic activity in the insect is consistent with the hypothesis that AY and other plant yellows diseases are caused by mycoplasma-like organisms.

  8. Molecular identification of aminoglycoside-modifying enzymes in clinical isolates of Escherichia coli resistant to amoxicillin/clavulanic acid isolated in Spain.

    Science.gov (United States)

    Fernández-Martínez, Marta; Miró, Elisenda; Ortega, Adriana; Bou, Germán; González-López, Juan José; Oliver, Antonio; Pascual, Alvaro; Cercenado, Emilia; Oteo, Jesús; Martínez-Martínez, Luis; Navarro, Ferran

    2015-08-01

    The activity of eight aminoglycosides (amikacin, apramycin, arbekacin, gentamicin, kanamycin, neomycin, netilmicin and tobramycin) against a collection of 257 amoxicillin/clavulanic acid (AMC)-resistant Escherichia coli isolates was determined by microdilution. Aminoglycoside resistance rates, the prevalence of aminoglycoside-modifying enzyme (AME) genes, the relationship between AME gene detection and resistance phenotype to aminoglycosides, and the association of AME genes with mechanisms of AMC resistance in E. coli isolates in Spain were investigated. Aminoglycoside-resistant isolates were screened for the presence of genes encoding common AMEs [aac(3)-Ia, aac(3)-IIa, aac(3)-IVa, aac(6')-Ib, ant(2″)-Ia, ant(4')-IIa and aph(3')-Ia] or 16S rRNA methylases (armA, rmtB, rmtC and npmA). In total, 105 isolates (40.9%) were resistant to at least one of the aminoglycosides tested. Amikacin, apramycin and arbekacin showed better activity, with MIC90 values of 2mg/L (arbekacin) and 8mg/L (amikacin and apramycin). Kanamycin presented the highest MIC90 (128mg/L). The most common AME gene was aac(6')-Ib (36 strains; 34.3%), followed by aph(3')-Ia (31 strains; 29.5%), ant(2″)-Ia (29 strains; 27.6%) and aac(3)-IIa (23 strains; 21.9%). aac(3)-Ia, aac(3)-IVa, ant(4')-IIa and the four methylases were not detected. The ant(2″)-Ia gene was usually associated with OXA-1 [21/30; 70%], whilst 23/25 (92%) strains producing CTX-M-15 had the aac(6')-Ib gene. The most prevalent AME gene was aac(6')-Ib (18/41; 44%) in nosocomial isolates, whilst ant(2″)-Ia and aph(3')-Ia genes (20/64; 31%) were more frequent in strains of community origin. In 64.6% isolates the phenotypic profile correlated with the presence of commonly encountered AMEs.

  9. Expression of Human Papillomavirus Type 16 L1 Protein in Transgenic Tobacco Plants

    Institute of Scientific and Technical Information of China (English)

    Hong-Li LIU; Wen-Sheng LI; Ting LEI; Jing ZHENG; Zheng ZHANG; Xiao-Fei YAN; Zhe-Zhi WANG; Yi-Li WANG; Lü-Sheng SI

    2005-01-01

    To develop a plant expression system for the production of the human papillomavirus type 16(HPV16) vaccine, we investigated whether the HPV16 L1 protein can be expressed in tobacco plants and whether it can be used as the cheapest form of edible vaccine. The HPV16 L1 coding sequence was amplified by PCR using specific primers from the plasmid pGEM-T-HPV16 containing the template sequence, and subcloned into the intermediate vector pUCmT and binary vector pBI121 consecutively to obtain the plant expression plasmid pBI-L1. The T-DNA regions of the pBI-L1 binary vector contained the constitutive Cauliflower mosaic virus (CaMV) 35S promoter and the neomycin phosphotransferase npt Ⅱ gene, which allowed the selection of transformed plants using kanamycin. The tobacco plants were transformed by cocultivating them, using the leaf disc method, with Agrobacterium tumefaciens LBA4404, which harbored the plant expression plasmid. The regenerated transgenic tobacco plants were selected using kanamycin, and confirmed by PCR. The results of the Southern blot assay also showed that the HPV16 L1 gene was integrated stably into the genome of the transformed tobacco plants. The Western blot analysis showed that the transformed tobacco leaves could express the HPV16 L1 protein. Furthermore, it was demonstrated by ELISA assay that the expressed protein accounted for 0.034%-0.076% of the total soluble leaf protein, was able to form 55 nm virus-like particles compatible with HPV virus-like particle (VLP), and induced mouse erythrocyte hemagglutination in vitro. The present results indicate that the HPV16 L1 protein can be expressed in transgenic tobacco plants and the expressed protein possesses the natural features of the HPV 16L1 protein, implying that the HPV16 L1 transgenic plants can be potentially used as an edible vaccine.

  10. 旋光法测定硫酸新霉素(饲料级)含量%Determination of neomycin sulfate in feed by polarimetry

    Institute of Scientific and Technical Information of China (English)

    卢芳; 刘群; 余祥华

    2006-01-01

    本文采用旋光法测定硫酸新霉素(饲料级)含量,结果表明,硫酸新霉素浓度为1500~15000 U/mL时,浓度与旋光度呈良好线性关系,相关系数r=0.9996(n=6).与微生物法相比,旋光法具有简便、快速、易操作等优点.

  11. [The effect of Solcoseryl eye gel in combination with neomycin and bacitracin on the healing of corneal lesions after foreign body removal].

    Science.gov (United States)

    Haydon, P

    1983-09-01

    The effect of Solcoseryl on the healing rate of corneal lesions after foreign body removal was assessed in a double-blind study. Solcoseryl is a protein-free hemodialysate which is widely used to stimulate tissue repair processes. The healing rate was determined by means of photodocumentation which was complete in 82 out of 92 cases. The Solcoseryl group showed in significantly faster healing rate and a lower tendency to development of corneal opacities than the control group. No side-effects were observed which could be attributed to either of the drugs used in the study.

  12. 曲利本蓝分光光度法测定新霉素的含量%Spectrophotometry of trypan blue determining the content of neomycin

    Institute of Scientific and Technical Information of China (English)

    湛海粼

    2003-01-01

    在pH2.0~6.0的条件下,曲利本蓝(TB)与硫酸新霉素(NEO)发生反应生成蓝色离子缔合物,最大显色波长位于680nm,最大褪色波长位于588nm,摩尔吸光系数(ε)分别为9.15×103和2.87×104L·mol-1·cm-1,线性范围在0~13.0μg/ml至0~16.0μg/ml之间.若用双波长叠加法测定,则ε值可达到3.79×104L·mol-1·cm-1.该方法用于实际样品的测定,结果满意.

  13. Determination of neomycin by derivatization with UV-spectrophotometry%衍生化紫外分光光度法测定新霉素含量

    Institute of Scientific and Technical Information of China (English)

    曾令高; 范能全; 曾杰; 刘映倩

    2006-01-01

    目的 建立硫酸新霉素中新霉素的含量测定方法.方法 以乙酰丙酮-甲醛为衍生化试剂的紫外分光光度法,检测波长为285nm.结果 新霉素在2.69~14.34μg·mL-1浓度范围内吸收度与浓度呈良好线性关系,r=0.999 7,平均回收率为98.63%.结论 该法准确,快速,结果满意.

  14. c-Myb knockdown increases the neomycin-induced damage to hair-cell-like HEI-OC1 cells in vitro

    OpenAIRE

    Xiaoyu Yu; Wenwen Liu; Zhaomin Fan; Fuping Qian; Daogong Zhang; Yuechen Han; Lei Xu; Gaoying Sun; Jieyu Qi; Shasha Zhang; Mingliang Tang; Jianfeng Li; Renjie Chai; Haibo Wang

    2017-01-01

    c-Myb is a transcription factor that plays a key role in cell proliferation, differentiation, and apoptosis. It has been reported that c-Myb is expressed within the chicken otic placode, but whether c-Myb exists in the mammalian cochlea, and how it exerts its effects, has not been explored yet. Here, we investigated the expression of c-Myb in the postnatal mouse cochlea and HEI-OC1 cells and found that c-Myb was expressed in the hair cells (HCs) of mouse cochlea as well as in cultured HEI-OC1...

  15. Determination of the relative amounts of the B and C components of neomycin by ion-exclusion chromatography using refractometric detection.

    Science.gov (United States)

    Decoster, W; Claes, P; Vanderhaeghe, H

    1981-07-03

    Refractometric detection can be used as a convenient alternative to ninhydrin colorimetric or polarimetric detection in ion-exclusion chromatography of neomycinn. The determination of the relative amounts of neomyci B and C using different detection methods is examined. The use of a resin of smaller granulometry and a medium-pressure chromatographic apparatus reduces the analysis time to less than 25 min.

  16. Rubber Tree (Hevea brasiliensis Muell. Arg).

    Science.gov (United States)

    Venkatachalam, Perumal; Jayashree, Radha; Rekha, Karumamkandathil; Sushmakumari, Sreedharannair; Sobha, Sankaren; Kumari Jayasree, Parukkuttyamma; Kala, Radha Gopikkuttanunithan; Thulaseedharan, Arjunan

    2006-01-01

    Rubber tree (Hevea brasiliensis Muell. Arg.) is an important industrial crop for natural rubber production. At present, more than 9.5 million hectares in about 40 countries are devoted to rubber tree cultivation with a production about 6.5 million tons of dry rubber each year. The world supply of natural rubber is barely keeping up with a global demand for 12 million tons of natural rubber in 2020. Tapping panel dryness (TPD) is a complex physiological syndrome widely found in rubber tree plantations, which causes severe yield and crop losses in natural rubber producing countries. Currently, there is no effective prevention or treatment for this serious malady. As it is a perennial tree crop, the integration of specific desired traits through conventional breeding is both time-consuming and labour-intensive. Genetic transformation with conventional breeding is certainly a more promising tool for incorporation of agronomically important genes that could improve existing Hevea genotype. This chapter provides an Agrobacterium-mediated transformation protocol for rubber tree using immature anther-derived calli as initial explants. We have applied this protocol to generate genetically engineered plants from a high yielding Indian clone RRII 105 of Hevea brasiliensis (Hb). Calli were co-cultured with Agrobacterium tumefaciens harboring a plasmid vector containing the Hb superoxide dismutase (SOD) gene and the reporter gene used was beta-glucuronidase (GUS) gene (uidA). The selectable marker gene used was neomycin phosphotransferase (nptII) and kanamycin was used as selection agent. We found that a suitable transformation protocol for Hevea consists of a 3-d co-cultivation with Agrobacterium in the presence of 20 mM acetosyringone, 15 mM betaine HCl, and 11.55 mM proline followed by selection on medium containing 300 mg/L kanamycin. Transformed calli surviving on medium containing 300 mg/L kanamycin showed a strong GUS-positive reaction. Upon subsequent subculture into

  17. 21 CFR 333.110 - First aid antibiotic active ingredients.

    Science.gov (United States)

    2010-04-01

    ... gram, 30 milligrams of chlortetracycline hydrochloride in a suitable ointment base. (d) Neomycin sulfate ointment containing, in each gram, 3.5 milligrams of neomycin in a suitable water soluble or oleaginous ointment base. (e) Neomycin sulfate cream containing, in each gram, 3.5 milligrams of neomycin...

  18. Occurrence of antibiotics in pharmaceutical industrial wastewater, wastewater treatment plant and sea waters in Tunisia.

    Science.gov (United States)

    Tahrani, Leyla; Van Loco, Joris; Ben Mansour, Hedi; Reyns, Tim

    2016-04-01

    Antibiotics are among the most commonly used group of pharmaceuticals in human medicine. They can therefore reach surface and groundwater bodies through different routes, such as wastewater treatment plant effluents, surface runoff, or infiltration of water used for agricultural purposes. It is well known that antibiotics pose a significant risk to environmental and human health, even at low concentrations. The aim of the present study was to evaluate the presence of aminoglycosides and phenicol antibiotics in municipal wastewaters, sea water and pharmaceutical effluents in Tunisia. All analysed water samples contained detectable levels of aminoglycoside and phenicol antibiotics. The highest concentrations in wastewater influents were observed for neomycin and kanamycin B (16.4 ng mL(-1) and 7.5 ng mL(-1), respectively). Chloramphenicol was found in wastewater influents up to 3 ng mL(-1). It was observed that the waste water treatment plants were not efficient in completely removing these antibiotics. Chloramphenicol and florfenicol were found in sea water samples near aquaculture sites at levels up to, respectively, 15.6 ng mL(-1) and 18.4 ng mL(-1). Also aminoglycoside antibiotics were found near aquaculture sites with the highest concentration of 3.4 ng mL(-1) for streptomycin. In pharmaceutical effluents, only gentamycin was found at concentrations up to 19 ng mL(-1) over a sampling period of four months.

  19. Associations of antimicrobial uses with antimicrobial resistance of fecal Escherichia coli from pigs on 47 farrow-to-finish farms in Ontario and British Columbia

    Science.gov (United States)

    Akwar, Holy T.; Poppe, Cornelis; Wilson, Jeff; Reid-Smith, Richard J.; Dyck, Monica; Waddington, Josh; Shang, Dayue; McEwen, Scott A.

    2008-01-01

    This study assessed the associations between antimicrobial use and other management practices in pigs and antimicrobial resistance in generic Escherichia coli recovered from feces of weaner and finisher pigs on 39 purposefully selected farrow-to-finish farms in Ontario and 8 in British Columbia. Antimicrobials (n = 13), most frequently penicillins and tetracycline, were administered to different age groups of pigs on study farms through various routes of administration. Logistic regression was used to model risk factors to antimicrobial resistance in fecal E. coli of pigs for the following antimicrobials: ampicillin, apramycin, carbadox, cephalothin, chloramphenicol, kanamycin, neomycin, nitrofurantoin, spectinomycin, streptomycin, sulfamethoxazole, tetracycline, and cotrimoxazole (trimethoprim and sulfamethoxazole). Use of antimicrobials in weaner pigs compared with use in finisher pigs was associated with resistance in most models. There was phenotypic evidence of different mechanisms of resistance selection, including direct selection [use of carbadox was associated with carbadox resistance (OR = 6.48)]; cross-resistance [use of spectinomycin was associated with streptomycin resistance (OR = 2.29)]; and possible co-selection [ceftiofur use was associated with tetracycline resistance (OR = 6.12)]. These results provide further evidence that use of antimicrobials in pigs selects for resistance among fecal E. coli within and between classes of antimicrobials. PMID:18505211

  20. Transferring cucumber mosaic virus-white leaf strain coat protein gene into Cucumis melo L. and evaluating transgenic plants for protection against infections

    Energy Technology Data Exchange (ETDEWEB)

    Gonsalves, C.; Xue, B.; Yepes, M.; Fuchs, M.; Ling, K.; Namba, S. (Cornell Univ., Geneva, NY (United States). Dept. of Plant Pathology)

    1994-03-01

    A single regeneration procedure using cotyledon examples effectively regenerated five commercially grown muskmelon cultivars. This regeneration scheme was used to facilitate gene transfers using either Agrobacterium tumefaciens or microprojectile bombardment methods. In both cases, the transferred genes were from the T-DNA region of the binary vector plasmid pGA482GG/cp cucumber mosaic virus-white leaf strain (CMV-WL), which contains genes that encode neomycin phosphotransferase II (NPT II), [beta]-glucuronidase (GUS), and the CMV-WL coat protein (CP). Explants treated with pGA482GG/cpCMV-WL regenerated shoots on Murashige and Skoog medium containing 4.4 [mu]m 6-benzylaminopurine (BA), kanamycin (Km) at 150 mg[center dot]liter[sup [minus]1] and carbenicillin (Cb) at 500 mg[center dot]liter[sup [minus]1]. The authors' comparison of A. tumefaciens- and microprojectile-mediated gene transfer procedures shows that both methods effectively produce nearly the same percentage of transgenic plants. R[sub 0] plants were first tested for GUS or NPT II expression, then the polymerase chain reaction (PCR) and other tests were used to verify the transfer of the NPT II, GUS, and CMV-WL CP genes.

  1. 氨基甙类抗生素的耳毒机制和细胞凋亡

    Institute of Scientific and Technical Information of China (English)

    王玉林; 董明敏

    2002-01-01

    @@ 自1944年Schatz等[1]报道链毒素(streptomycin,SM)的抗菌活性以来,氨基甙类抗生素(aminoglycoside antibiotics,AmAn)逐渐变成世界范围内应用最广泛的抗生素品种之一,如卡那霉素(kanamycin,KM)、庆大霉素(gentamycin,GM)、新霉素(neomycin)及相关的新合成或半合成的AmAn的出现和应用.为治疗威胁人类生命的疾病做出了巨大贡献. AmAn的基本结构是已糖环,由2个或2个以上的氨基糖通过糖甙键与之结合形成化学稳定的复合物,一般称氨基糖甙,根据氨基糖的不同,对AmAn进行分类.AmAn的共同特点为:(1)水溶性好,性质稳定.(2)抗菌谱广,对葡萄球菌属、需氧革兰氏阴性杆菌均具有良好的抗菌活性.(3)抗菌机制主要是杀菌作用,抑制细菌蛋白质的合成.

  2. Detection of antibiotic residues in bovine milk by a voltammetric electronic tongue system.

    Science.gov (United States)

    Wei, Zhenbo; Wang, Jun

    2011-05-23

    A voltammetric electronic tongue (VE-tongue) was developed to detect antibiotic residues in bovine milk. Six antibiotics (Chloramphenicol, Erythromycin, Kanamycin sulfate, Neomycin sulfate, Streptomycin sulfate and Tetracycline HCl) spiked at four different concentration levels (0.5, 1, 1.5 and 2 maximum residue limits (MRLs)) were classified based on VE-tongue by two pattern recognition methods: principal component analysis (PCA) and discriminant function analysis (DFA). The VE-tongue was composed of five working electrodes (gold, silver, platinum, palladium, and titanium) positioned in a standard three-electrode configuration. The Multi-frequency large amplitude pulse voltammetry (MLAPV) which consisted of four segments (1 Hz, 10 Hz, 100 Hz and 1000 Hz) was applied as potential waveform. The six antibiotics at the MRLs could not be separated from bovine milk completely by PCA, but all the samples were demarcated clearly by DFA. Three regression models: Principal Component Regression Analysis (PCR), Partial Least Squares Regression (PLSR), and Least Squares-Support Vector Machines (LS-SVM) were used for concentrations of antibiotics prediction. All the regression models performed well, and PCR had the most stable results.

  3. Crystallization and preliminary crystallographic analysis of hygromycin B phosphotransferase from Escherichia coli

    Energy Technology Data Exchange (ETDEWEB)

    Iino, Daisuke [Department of Bioscience, Tokyo University of Agriculture, Tokyo 156-8502 (Japan); Takakura, Yasuaki [Division of Integrative Environmental Sciences, Graduate School of Life and Environmental Sciences, University of Tsukuba, Ibaraki 305-8572 (Japan); Kuroiwa, Mika; Kawakami, Ryouta; Sasaki, Yasuyuki [Department of Bioscience, Tokyo University of Agriculture, Tokyo 156-8502 (Japan); Hoshino, Takayuki [Division of Integrative Environmental Sciences, Graduate School of Life and Environmental Sciences, University of Tsukuba, Ibaraki 305-8572 (Japan); Ohsawa, Kanju [Department of Bioscience, Tokyo University of Agriculture, Tokyo 156-8502 (Japan); Nakamura, Akira [Division of Integrative Environmental Sciences, Graduate School of Life and Environmental Sciences, University of Tsukuba, Ibaraki 305-8572 (Japan); Yajima, Shunsuke, E-mail: yshun@nodai.ac.jp [Department of Bioscience, Tokyo University of Agriculture, Tokyo 156-8502 (Japan)

    2007-08-01

    The crystallization and preliminary X-ray studies of the aminoglycoside antibiotic-modifying enzyme hygromycin B phosphotransferase from E. coli are reported. Aminoglycoside antibiotics, such as hygromycin, kanamycin, neomycin, spectinomycin and streptomycin, inhibit protein synthesis by acting on bacterial and eukaryotic ribosomes. Hygromycin B phosphotransferase (Hph; EC 2.7.1.119) converts hygromycin B to 7′′-O-phosphohygromycin using a phosphate moiety from ATP, resulting in the loss of its cell-killing activity. The Hph protein has been crystallized for the first time using a thermostable mutant and the hanging-drop vapour-diffusion method. The crystal provided diffraction data to a resolution of 2.1 Å and belongs to space group P3{sub 2}21, with unit-cell parameters a = b = 71.0, c = 125.0 Å. Crystals of complexes of Hph with hygromycin B and AMP-PNP or ADP have also been obtained in the same crystal form as that of the apoprotein.

  4. Surface plasmon resonance analysis of antibiotics using imprinted boronic acid-functionalized Au nanoparticle composites.

    Science.gov (United States)

    Frasconi, Marco; Tel-Vered, Ran; Riskin, Michael; Willner, Itamar

    2010-03-15

    Au nanoparticles (NPs) are functionalized with thioaniline electropolymerizable groups and (mercaptophenyl)boronic acid. The antibiotic substrates neomycin (NE), kanamycin (KA), and streptomycin (ST) include vicinal diol functionalities and, thus, bind to the boronic acid ligands. The electropolymerization of the functionalized Au NPs in the presence of NE, KA, or ST onto Au surfaces yields bisaniline-cross-linked Au NP composites that, after removal of the ligated antibiotics, provide molecularly imprinted matrixes which reveal high sensitivities toward the sensing of the imprinted antibiotic analytes (detection limits for analyzing NE, KA, and ST correspond to 2.00 +/- 0.21 pM, 1.00 +/- 0.10 pM, and 200 +/- 30 fM, respectively). The antibiotics are sensed by surface plasmon resonance (SPR) spectroscopy, where the coupling between the localized plasmon of the NPs and the surface plasmon wave associated with the Au surface is implemented to amplify the SPR responses. The imprinted Au NP composites are, then, used to analyze the antibiotics in milk samples.

  5. 鸭疫里氏杆菌的分离鉴定及药敏试验%Isolation, Identification and Drug-sensitivity Tests of Riemerella anatipestifer Isolates

    Institute of Scientific and Technical Information of China (English)

    潘淑惠; 王璇; 郭光容; 徐景峨; 吴位珩; 杨莉; 余波

    2011-01-01

    为鸭疫里氏杆菌病的防治提供参考,2009年5月,对贵州某肉鸭养殖场送检病鸭进行了剖检病变观察、细菌分离鉴定,并进行了药敏试验.结果表明,分离细菌通过细菌形态、染色特性、生化试验、培养特征鉴定为鸭疫里氏杆菌,7株菌对头孢拉定、头孢曲松钠、头孢噻肟和氟苯尼考高度敏感,对庆大霉素、新霉素、青霉素、多粘菌素B、氟哌酸、复方新诺明、链霉素和卡那霉素耐药.%To make references for prevention and treatment of Riemerella anatipestifer, in May 2009, pathological changes of sick duck was observed, bacteria were isolated and identified and drug-sensitivity tests were carried out in Guizhou meat duck farm. The results showed that the isolated bacteria were identified as Riemerella anatipestifer by bacterium form, biochemistry and cultured characteristics. The seven strains were highly susceptible to cefradine, ceftriaxone sodium, cefotaxime, florfenicol, not susceptible to gentamicin, neomycin, penicillin, polymyxin B, norfloxacin, sinomin compositea, streptomycin and kanamycin.

  6. Detection of antibiotic residues in bovine milk by a voltammetric electronic tongue system

    Energy Technology Data Exchange (ETDEWEB)

    Wei Zhenbo [Zhejiang University, Department of Bio-Systems Engineering, 268 Kaixuan Road, Hangzhou, Zhejiang 310029 (China); Wang Jun, E-mail: jwang@zju.edu.cn [Zhejiang University, Department of Bio-Systems Engineering, 268 Kaixuan Road, Hangzhou, Zhejiang 310029 (China)

    2011-05-23

    A voltammetric electronic tongue (VE-tongue) was developed to detect antibiotic residues in bovine milk. Six antibiotics (Chloramphenicol, Erythromycin, Kanamycin sulfate, Neomycin sulfate, Streptomycin sulfate and Tetracycline HCl) spiked at four different concentration levels (0.5, 1, 1.5 and 2 maximum residue limits (MRLs)) were classified based on VE-tongue by two pattern recognition methods: principal component analysis (PCA) and discriminant function analysis (DFA). The VE-tongue was composed of five working electrodes (gold, silver, platinum, palladium, and titanium) positioned in a standard three-electrode configuration. The Multi-frequency large amplitude pulse voltammetry (MLAPV) which consisted of four segments (1 Hz, 10 Hz, 100 Hz and 1000 Hz) was applied as potential waveform. The six antibiotics at the MRLs could not be separated from bovine milk completely by PCA, but all the samples were demarcated clearly by DFA. Three regression models: Principal Component Regression Analysis (PCR), Partial Least Squares Regression (PLSR), and Least Squares-Support Vector Machines (LS-SVM) were used for concentrations of antibiotics prediction. All the regression models performed well, and PCR had the most stable results.

  7. Antimicrobial susceptibility pattern of Edwardsiella ictaluri isolates from natural outbreaks of bacillary necrosis of Pangasianodon hypophthalmus in Vietnam.

    Science.gov (United States)

    Tu, Thanh Dung; Haesebrouck, Freddy; Nguyen, Anh Tuan; Sorgeloos, Patrick; Baele, Margo; Decostere, Annemie

    2008-12-01

    The purpose of this study was to assess the in vitro susceptibility of 64 Vietnamese isolates of Edwardsiella ictaluri, the causal agent of the infectious disease Bacillus Necrosis Pangasius in Pangasianodon hypophthalmus, using the agar dilution technique. All isolates originated from different farms and were collected between 2002 and 2005. None of the isolates displayed acquired resistance to amoxicillin, amoxicillin-clavulanic acid, chloramphenicol, florfenicol, gentamicin, kanamycin, neomycin, and nitrofurantoin. Acquired resistance to streptomycin was detected in 83%, to oxytetracycline in 81%, and to trimethoprim in 71% of the isolates, as indicated by a bimodal distribution of the minimal inhibitory concentrations (MICs) of these antimicrobials. The MICs of enrofloxacin displayed a monomodal distribution with tailing toward the higher MIC values, possibly indicating reduced susceptibility of a minority of isolates (3 out of the 64). For the quinolone antimicrobial agents flumequin and oxolinic acid, acquired resistance was encountered in 8% and 6% of the strains, respectively. All strains were intrinsically resistant to the polypeptide antimicrobial agent colistin. Seventy-three percent of the isolates were shown to have acquired resistance to at least three antimicrobial agents. The results of this study emphasize the strict need to control both the prophylactic and curative use of antimicrobial agents in Vietnamese aquaculture.

  8. Quantifying Attachment and Antibiotic Resistance of from Conventional and Organic Swine Manure.

    Science.gov (United States)

    Zwonitzer, Martha R; Soupir, Michelle L; Jarboe, Laura R; Smith, Douglas R

    2016-03-01

    Broad-spectrum antibiotics are often administered to swine, contributing to the occurrence of antibiotic-resistant bacteria in their manure. During land application, the bacteria in swine manure preferentially attach to particles in the soil, affecting their transport in overland flow. However, a quantitative understanding of these attachment mechanisms is lacking, and their relationship to antibiotic resistance is unknown. The objective of this study is to examine the relationships between antibiotic resistance and attachment to very fine silica sand in collected from swine manure. A total of 556 isolates were collected from six farms, two organic and four conventional (antibiotics fed prophylactically). Antibiotic resistance was quantified using 13 antibiotics at three minimum inhibitory concentrations: resistant, intermediate, and susceptible. Of the 556 isolates used in the antibiotic resistance assays, 491 were subjected to an attachment assay. Results show that isolates from conventional systems were significantly more resistant to amoxicillin, ampicillin, chlortetracycline, erythromycin, kanamycin, neomycin, streptomycin, tetracycline, and tylosin ( < 0.001). Results also indicate that isolated from conventional systems attached to very fine silica sand at significantly higher levels than those from organic systems ( < 0.001). Statistical analysis showed that a significant relationship did not exist between antibiotic resistance levels and attachment in from conventional systems but did for organic systems ( < 0.001). Better quantification of these relationships is critical to understanding the behavior of in the environment and preventing exposure of human populations to antibiotic-resistant bacteria.

  9. A mutation in the 530 loop of Escherichia coli 16S ribosomal RNA causes resistance to streptomycin.

    Science.gov (United States)

    Melançon, P; Lemieux, C; Brakier-Gingras, L

    1988-10-25

    Oligonucleotide-directed mutagenesis was used to introduce an A to C transversion at position 523 in the 16S ribosomal RNA gene of Escherichia coli rrnB operon cloned in plasmid pKK3535. E. coli cells transformed with the mutated plasmid were resistant to streptomycin. The mutated ribosomes isolated from these cells were not stimulated by streptomycin to misread the message in a poly(U)-directed assay. They were also restrictive to the stimulation of misreading by other error-promoting related aminoglycoside antibiotics such as neomycin, kanamycin or gentamicin, which do not compete for the streptomycin binding site. The 530 loop where the mutation in the 16S rRNA is located has been mapped at the external surface of the 30S subunit, and is therefore distal from the streptomycin binding site at the subunit interface. Our results support the conclusion that the mutation at position 523 in the 16S rRNA does not interfere with the binding of streptomycin, but prevents the drug from inducing conformational changes in the 530 loop which account for its miscoding effect. Since this effect primarily results from a perturbation of the translational proofreading control, our results also provide evidence that the 530 loop of the 16S rRNA is involved in this accuracy control.

  10. A new rapid and sensitive bioluminescence assay for antibiotics that inhibit protein synthesis.

    Science.gov (United States)

    Naveh, A; Potasman, I; Bassan, H; Ulitzur, S

    1984-06-01

    A new sensitive, rapid and simple bioluminescence assay for antibiotics inhibiting protein synthesis is described. In this assay the ability of the tested antibiotic to inhibit the de novo synthesis of the enzymes participating in the bacterial luminescence system is determined by means of a dark variant of a luminous bacterium that undergoes prompt induction of the luminescence system with certain DNA-intercalating agents. Upon induction, the in vivo luminescence of the dark variant is increased more than 50-fold within 30 min. Antibiotics that block the de novo synthesis of protein limit the development of luminescence at a level that was found to be a function of the antibiotic concentration. The minimum detectable concentration of antibiotics in the bioluminescence test, after 45-60 min of incubation, was 0.1 microgram/ml for streptomycin, gentamicin, kanamycin, lincomycin and chloramphenicol and 0.3 microgram/ml for neomycin, clindamycin and spectinomycin. The new bioluminescence test has been used to assay these antibiotics in serum.

  11. Stable genetic transformation of Vigna mungo L. Hepper via Agrobacterium tumefaciens.

    Science.gov (United States)

    Saini, R; Sonia; Jaiwal, P K; Jaiwal, S

    2003-06-01

    Vigna mungo is one of the large-seeded grain legumes that has not yet been transformed. We report here for the first time the production of morphologically normal and fertile transgenic plants from cotyledonary-node explants inoculated with Agrobacterium tumefaciens carrying binary vector pCAMBIA2301, the latter of which contains a neomycin phosphotransferase ( nptII) gene and a beta-glucuronidase (GUS) gene ( uidA) interrupted with an intron. The transformed green shoots, selected and rooted on medium containing kanamycin, tested positive for nptII and uidA genes by polymerase chain reaction (PCR) analysis. These shoots were established in soil and grown to maturity to collect the seeds. Mechanical wounding of the explants prior to inoculation with Agrobacterium, time lag in regeneration due to removal of the cotyledons from explants and a second round of selection at the rooting stage were found to be critical for transformation. Analysis of T(0) plants showed the expression and integration of uidA into the plant genome. GUS activity in leaves, roots, flowers, anthers and pollen grains was detected by histochemical assay. PCR analysis of T(1) progeny revealed a Mendelian transgene inheritance pattern. The transformation frequency was 1%, and 6-8 weeks were required for the generation of transgenics.

  12. Transformation of a recalcitrant grain legume, Vigna mungo L. Hepper, using Agrobacterium tumefaciens-mediated gene transfer to shoot apical meristem cultures.

    Science.gov (United States)

    Saini, Raman; Jaiwal, Pawan K

    2005-06-01

    The efficiency of Vigna mungo L. Hepper transformation was significantly increased from an average of 1% to 6.5% by using shoot apices excised from embryonic axes precultured on 10 microM benzyl-6-aminopurine (BAP) for 3 days and wounded prior to inoculation in Agrobacterium tumefaciens strain EHA105 carrying the binary vector pCAMBIA2301, which contains a neomycin phosphotransferase gene (nptII) and a beta-glucuronidase (GUS) gene (gusA) interrupted by an intron. The transformed green shoots that were selected and rooted on medium containing kanamycin, and which tested positive for nptII gene by polymerase chain reaction, were established in soil to collect seeds. GUS activity was detected in whole T(0) shoots and T(1) seedlings. All T(0) plants were morphologically normal, fertile and the majority of them transmitted transgenes in a 3:1 ratio to their progenies. Southern analysis of T(1) plants showed integration of nptII into the plant genome.

  13. Transformation of blackgram (Vigna mungo (L.) Hepper) by barley chitinase and ribosome-inactivating protein genes towards improving resistance to Corynespora leaf spot fungal disease.

    Science.gov (United States)

    Chopra, Rajan; Saini, Raman

    2014-12-01

    Blackgram (Vigna mungo (L.) Hepper), an important grain legume crop, is sensitive to many fungal pathogens including Corynespora cassiicola, the causal agent of corynespora leaf spot disease. In the present study, plasmid pGJ42 harboring neomycin phosphotransferase (nptII) a selectable marker gene, the barley antifungal genes chitinase (AAA56786) and ribosome-inactivating protein (RIP; AAA32951) were used for the transformation, to develop fungal resistance for the first time in blackgram. The presence and integration of transgene into the blackgram genome was confirmed by PCR and Southern analysis with an overall transformation frequency of 10.2 %. Kanamycin selection and PCR analysis of T0 progeny revealed the inheritance of transgene in Mendelian fashion (3:1). Transgenic plants (T1), evaluated for fungal resistance by in vitro antifungal assay, arrested the growth of C. cassiicola up to 25-40 % over the wild-type plants. In fungal bio-assay screening, the transgenic plants (T1) sprayed with C. cassiicola spores showed a delay in onset of disease along with their lesser extent in terms of average number of diseased leaves and reduced number and size of lesions. The percent disease protection among different transformed lines varies in the range of 27-47 % compare to control (untransformed) plants. These results demonstrate potentiality of chitinase and RIP from a heterologous source in developing fungal disease protection in blackgram and can be helpful in increasing the production of blackgram.

  14. A versatile binary vector system with a T-DNA organisational structure conducive to efficient integration of cloned DNA into the plant genome.

    Science.gov (United States)

    Gleave, A P

    1992-12-01

    A versatile gene expression cartridge and binary vector system was constructed for use in Agrobacterium-mediated plant transformation. The expression cartridge of the primary cloning vector, pART7, comprises of cauliflower mosaic virus Cabb B-JI isolate 35S promoter, a multiple cloning site and the transcriptional termination region of the octopine synthase gene. The entire cartridge can be removed from pART7 as a Not I fragment and introduced directly into the binary vector, pART27, recombinants being selected by blue/white screening for beta-galactosidase. pART27 carries the RK2 minimal replicon for maintenance in Agrobacterium, the ColE1 origin of replication for high-copy maintenance in Escherichia coli and the Tn7 spectinomycin/streptomycin resistance gene as a bacterial selectable marker. The organisational structure of the T-DNA of pART27 has been constructed taking into account the right to left border, 5' to 3' model of T-DNA transfer. The T-DNA carries the chimaeric kanamycin resistance gene (nopaline synthase promoter-neomycin phosphotransferase-nopaline synthase terminator) distal to the right border relative to the lacZ' region. Utilisation of these vectors in Agrobacterium-mediated transformation of tobacco demonstrated efficient T-DNA transfer to the plant genome.

  15. Prevalence, molecular characterization and antimicrobial resistance of Salmonella serovars isolated from northwestern Spanish broiler flocks (2011-2015).

    Science.gov (United States)

    Lamas, A; Fernandez-No, I C; Miranda, J M; Vázquez, B; Cepeda, A; Franco, C M

    2016-09-01

    The present study investigated the prevalence, antimicrobial resistance to twenty antibiotics, and class 1 integron and virulence genes of Salmonella isolated from poultry houses of broilers in northwestern Spain between 2011 and 2015. Strains were classified to the serotype level using the Kauffman-White typing scheme and subtyping with enterobacterial repetitive intergenic consensus PCR. The prevalence of Salmonella spp. was 1.02%. Sixteen different serotypes were found, with S. typhimurium and S. arizonae 48:z4, z23:- being the most prevalent. A total of 59.70% of strains were resistant to at least one, and 19.70% were resistant to multiple drugs. All Salmonella spp. were susceptible to cefotaxime, ciprofloxacin, gentamicin, kanamycin, levofloxacin, neomycin, and trimethoprim. The highest level of resistance was to sulfamethoxazole (40.29%), doxycycline (17.91%), and nalidixic acid (17.91%). None of the isolates carried class 1 integron and only isolates of S. enterica subspecies enterica were positive for all virulence factors tested, whereas S. arizonae lacked genes related to replication and invasion in nonphagocytic cells. This study demonstrates that the prevalence and antimicrobial resistance of Salmonella spp. in poultry houses of broilers of northwestern Spain is low compared with those found in other studies and in other steps of the food chain. © 2016 Poultry Science Association Inc.

  16. A survey of the frequency of aminoglycoside antibiotic-resistant genotypes and phenotypes in Escherichia coli in broilers with septicaemia in Hebei, China.

    Science.gov (United States)

    Zhang, F Y; Huo, S Y; Li, Y R; Xie, R; Wu, X J; Chen, L G; Gao, Y H

    2014-01-01

    1. The aim of this study was to investigate the occurrence of aminoglycoside resistance and the prevalence of 6 important modifying enzyme genes, i.e. (strA, strB, aph(3')-IIa, aac(3)-IIa, aac(6')-Ib and ant(3")-Ia), in Escherichia coli strains in broilers with septicaemia in Hebei, China. 2. A total of 111 clinical isolates of E. coli were collected from 46 large-scale farms. Antimicrobial susceptibility tests, using the Kirby-Bauer disc diffusion method, were performed on all 111 isolates. In addition, all were screened for the presence of modifying enzyme genes using the polymerase chain reaction (PCR). 3. The results show that the rates of resistance were as follows: streptomycin: 97.3%, kanamycin: 97.0%, gentamicin: 95.5%, neomycin: 50.5%, amikacin: 46.0%, spectinomycin: 22.5%. Of the genes examined, strB (73.9%) was the most frequently identified gene in the phenotypic resistant isolates, followed in order by: ant(3")-Ia, aac(3)-IIa, aac(6')-Ib, aph(3')-IIa and strA. 4. It is concluded that aminoglycoside resistance in E. coli from broilers with septicaemia remains a serious problem in Hebei, China. This emphasises the need to ban the non-therapeutic use of antibiotics, discourage their misuse and to be continually vigilant by providing appropriate scientific and technological support for the poultry industry.

  17. Use of lambda Red-mediated recombineering and Cre/lox for generation of markerless chromosomal deletions in avian pathogenic Escherichia coli.

    Science.gov (United States)

    Tuntufye, Huruma N; Goddeeris, Bruno M

    2011-12-01

    Avian pathogenic Escherichia coli (APEC) are bacteria associated with extraintestinal diseases in poultry. A method to generate markerless deletions of APEC genome is described. Lambda Red recombination is used to introduce a LoxP cassette (loxP-rpsL-neo-loxP) containing the rpsL gene for streptomycin sensitivity and the neo gene for kanamycin/neomycin resistance into the APEC genome, with attendant deletion of a desired chromosomal gene. The loxP sites are incorporated into primers used to amplify the rpsL-neo marker during the construction of the LoxP cassette, making the method rapid and efficient. The cassette is specifically integrated into the fiu gene or intergenic region 2051-52, and the Cre/lox system is used to remove the marker, hence deletion of the drug-resistance genes. The results demonstrate that the Cre/lox system can successfully be used to generate markerless deletions in APEC, and rpsL counter-selection can be used to select the deletions so that one does not have to pick and test to find the desired product. © 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

  18. Occurrence of antimicrobial residues in Brazilian food animals in 2008 and 2009.

    Science.gov (United States)

    Nonaka, C K V; Oliveira, A M G; Paiva, C R; Almeida, M P; Rezende, C P; Moraes, C G O; Botelho, B G; Souza, L F; Dias, P G

    2012-01-01

    Brazil is one of the most important countries as a producer and exporter of cattle and poultry. In 2009 cattle accounted for 30% of the export market and 41.4% for poultry meat. The Brazilian National Residues and Contaminants Control Plan (PNCRC) follows the guidelines set by the Codex Alimentarius Commission and checks compliance maximum residue limits (MRLs) to ensure the quality of these commodities. Kidney samples (n = 2978) were analysed between January 2008 and December 2009. Fifteen antibiotics of the macrolide and aminoglycoside groups (clindamycin, eritromycin, lincomycin, tylmicosin, tylosin, amikacin, apramycin, dihydrostreptomycin, gentamycin, higromycin, kanamycin, neomycin, spectinomycin, streptomycin, tobramycin) were determined by a microbiological screening method (FAST) and confirmed/quantified using liquid chromatography (LC-MS/MS and UPLC-MS/MS). In 2008, 1459 samples were analysed by a screening test and liquid chromatography with only one sample (0.07%) exceeded Brazilian legislation limits (>MRL). In 2009, 1519 samples were analysed and none exceeding Brazilian legislation limits (>MRL). The slaughterhouses of 16 states were monitored during the year of 2008, and 18 states were monitored in 2009, being the major producing states most sampled by the PNCRC.

  19. Simultaneous determination of 15 aminoglycoside(s) residues in animal derived foods by automated solid-phase extraction and liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Tao, Yanfei; Chen, Dongmei; Yu, Huan; Huang, Lingli; Liu, Zhaoying; Cao, Xiaoqin; Yan, Caixia; Pan, Yuanhu; Liu, Zhenli; Yuan, Zonghui

    2012-11-15

    An automated method has been developed for the simultaneous quantification of 15 aminoglycosides in muscle, liver (pigs, chicken and cattle), kidney (pigs and cattle), cow milk, and hen eggs by liquid chromatography tandem mass spectrometry. Homogenized samples were extracted by monopotassium phosphate buffer (including ethylene diamine tetraacetic acid), and cleaned up with auto solid-phase extraction by carboxylic acid cartridges. The analytes were separated by a specialized column for aminoglycosides, and eluted with trifluoroacetic acid and acetonitrile. The decision limits (CCα) of apramycin, gentamycin, tobramycin, paromomycin, hygromycin, neomycin, kanamycin, sisomicin, netilmicin, ribostamycin, kasugamycin, amikacin, streptomycin, dihydrostreptomycin and spectinomycin were ranged from 8.1 to 11.8 μg/kg and detection capabilities (CCβ) from 16.4 to 21.8 μg/kg. High correlation coefficients (r(2)>0.99) of calibration curves for the analytes were obtained within linear from 20 to 1000 μg/kg. Reasonable recoveries (71-108%) were demonstrated with excellent relative standard deviation (RSD). This method is simple pretreatment, rapid determination and high sensitivity, which can be used in the determination of multi-aminoglycosides in complex samples. Copyright © 2012 Elsevier Ltd. All rights reserved.

  20. Cultural characteristics of chromium resistant unicellular cyanobacteria isolated from local environment in Pakistan

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    Many unicellular cyanobacteria were isolated from different places: fields, ponds, polluted water, and soils from Muredkey and Kasur tannery areas, near Lahore, Pakistan. Different media like BG 11 medium, Bold Basal medium, Chu's # 10 medium and Gorham's medium, in standard forms and with slight variations of ingredients, and different pH, temperature and light regimes were checked for the optimum growth of the isolates. The isolation procedure was repeated with different concentrations of chromium to select the resistant strains. These selected strains grew on chromium of the range 100-200 μg/ml in BG 11 medium. Cyanobacteria were maintained in solid and liquid media with/without shaking. Cyanobacterial strains were collected from natural habitats that were accompanied by a diversified group of organisms including bacteria, protozoan, and rotifers etc. In order to eliminate these agents termed as contaminants, we used several methods including phenol treatment, use of antibiotic and careful manual picking of unicellular cyanobacteria. Resistance of these strains against different heavy metals (ZnSO4, MnSO4,NiSO4, CoCl2, Pb(NO3)3, CuSO4, HgCl2, AgNO3 and CdCl2) and antibiotics (erythromycin, streptomycin, kanamycin,chloramphenicol, neomycin) was evaluated. Optimum temperature was 30℃ with variable pH for the reduction of Cr6+ in to Cr3+ in majority of strains.

  1. Genetic transformation of mature embryos of bread (T. aestivum) and pasta (T. durum) wheat genotypes.

    Science.gov (United States)

    Moghaieb, Reda E A; El-Arabi, Nagwa I; Momtaz, Osama A; Youssef, Sawsan S; Soliman, Mohamed H

    2010-01-01

    The objective of the present study is to develop an efficient protocol for regeneration of transgenic wheat plants using Agrobacterium- mediated transformation of mature embryos of hexaploid bread wheat (Triticum aestivum) and tetraploid pasta wheat (Triticum durum). The data indicated that embryogenic calli were formed within 7 days in the presence of 2 mgl-1 2,4-D. Adventitious shoots emerged from the embryonic calli in the presence of 2 mgl-1 BA. Shoot regeneration frequency varied between wheat cultivars according to their genetic background differences. Regeneration frequency was higher in the cultivar Gemmiza 10 (95 %) compared with the other cultivars tested. Mature embryos derived callus of the cultivars Gemmiza 10 and Gemmiza 9 were co-cultivated with A. tumefaciens strain LBA4404 harboring a binary vector pBI-121 containing the neomycin phosphotransferase-II gene (npt-II). The resulted putative transgenic plantlets were able to grow on kanamycin containing medium. A successful integration of the transgene was confirmed by analyzing the T0 plantlets using Southern hybridization and PCR amplification. The gus gene expression can be detected only in the transgenic plants. The reported protocol is reproducible and can be used to regenerate transgenic wheat plants expressing the genes present in A. tumifaciens binary vectors.

  2. PFGE, Lior serotype, and antimicrobial resistance patterns among Campylobacter jejuni isolated from travelers and US military personnel with acute diarrhea in Thailand, 1998-2003

    Directory of Open Access Journals (Sweden)

    Serichantalergs Oralak

    2010-11-01

    Full Text Available Abstract Background Campylobacter jejuni is a major cause of gastroenteritis worldwide. In Thailand, several strains of C. jejuni have been isolated and identified as major diarrheal pathogens among adult travelers. To study the epidemiology of C. jejuni in adult travelers and U.S. military personnel with acute diarrhea in Thailand from 1998-2003, strains of C. jejuni were isolated and phenotypically identified, serotyped, tested for antimicrobial susceptibility, and characterized using pulsed-field gel electrophoresis (PFGE. Results A total of 312 C. jejuni isolates were obtained from travelers (n = 46 and U.S. military personnel (n = 266 in Thailand who were experiencing acute diarrhea. Nalidixic acid and ciprofloxacin resistance was observed in 94.9% and 93.0% of the isolates, respectively. From 2001-2003, resistance to tetracycline (81.9%, trimethoprim-sulfamethoxazole (57.9%, ampicillin (28.9%, kanamycin (5.9%, sulfisoxazole (3.9%, neomycin (2.0%, and streptomycin (0.7% was observed. Combined PFGE analysis showed considerable genetic diversity among the C. jejuni isolates; however, four PFGE clusters included isolates from the major Lior serotypes (HL: 36, HL: 11, HL: 5, and HL: 28. The PFGE analysis linked individual C. jejuni clones that were obtained at U.S. military exercises with specific antimicrobial resistance patterns. Conclusions In summary, most human C. jejuni isolates from Thailand were multi-resistant to quinolones and tetracycline. PFGE detected spatial and temporal C. jejuni clonality responsible for the common sources of Campylobacter gastroenteritis.

  3. Transformation of soybean protoplasts from permanent suspension cultures by cocultivation with cells of Agrobacterium tumefaciens.

    Science.gov (United States)

    Baldes, R; Moos, M; Geider, K

    1987-03-01

    Cell wall regenerating protoplasts from soybean cells kept in suspension culture were cocultivated with bacteria which were derived from the nopaline strain C58 of Agrobacterium tumefaciens. When the bacteria carried an oncogenic Ti-plasmid, about 5% of the surviving protoplasts were able to form calli on hormone-free agar in contrast to controls, where bacteria without Ti-plasmid were applied, and where no calli were formed. After isolation of DNA from hormone-independently growing cells further evidence for transformation was obtained by hybridization to Ti-plasmid specific RNA and by rescue of a segment with a bacterial resistance gene which had been inserted before into the T-DNA. Transfer of T-DNA harboring a neomycin-resistance gene activated by the nos-promoter resulted in calli growing on kanamycin. Verification of segments located at the left and the right part of the T-DNA indicated the presence of its entire length in transformed soybean cells. Expression of T-DNA genes was measured by the assay of nopaline-synthase. Cells cultured on agar had a much higher level of nopaline-synthase than fast growing cells in suspension culture. Transferring them to agar or treatment with azacytidine strongly increased synthesis of nopaline-synthase indicating a reversible repression presumably via a methylation mechanism.

  4. In vitro activity of flumequine in comparison with several other antimicrobial agents against five pathogens isolated in calves in The Netherlands.

    Science.gov (United States)

    Mevius, D J; Breukink, H J; van Miert, A S

    1990-10-01

    The in vitro activity of flumequine in comparison with several other drugs was tested against 17 P. multocida, 16 P. haemolytica, 21 S. dublin, 21 S. typhimurium and 21 E. coli strains, isolated in (veal) calves in the Netherlands. The MIC50 of flumequine for the respective pasteurellas was 0.25 and 1 microgram/ml, for the salmonellas and E. coli 0.5 micrograms/ml. In comparison with flumequine, enrofloxacin and ciprofloxacin showed higher in vitro activity, with MIC50 less than or equal to 0.008 micrograms/ml for ciprofloxacin. Decreased susceptibility of the pasteurellas was found for kanamycin, neomycin, streptomycin, gentamicin, oxytetracycline and doxycycline. The MIC50 of minocycline for P. multocida was 0.5 micrograms/ml and there was no cross resistance with the other tetracyclines. P. multocida was very susceptible to ampicillin (MIC50 less than or equal to 0.03 micrograms/ml), P. haemolytica, however, was 100% resistant to this drug. Both pasteurellas were susceptible to cephalothin and approximately 50% of the strains of both bacteria were resistant to chloramphenicol. The MIC50 of either spiramycin or tylosin was greater than or equal to their respective breakpoint-MIC values. Both pasteurellas were susceptible to the combination of trimethoprim and sulphamethoxazole. However, for P. multocida, the addition of sulphamethoxazole to trimethoprim had no synergistic effect on its MIC. In comparison with trimethorpim, aditoprim was less potent. Therefore only P. multocida was susceptible to aditoprim.

  5. A method of batch-purifying microalgae with multiple antibiotics at extremely high concentrations

    Science.gov (United States)

    Han, Jichang; Wang, Song; Zhang, Lin; Yang, Guanpin; Zhao, Lu; Pan, Kehou

    2016-01-01

    Axenic microalgal strains are highly valued in diverse microalgal studies and applications. Antibiotics, alone or in combination, are often used to avoid bacterial contamination during microalgal isolation and culture. In our preliminary trials, we found that many microalgae ceased growing in antibiotics at extremely high concentrations but could resume growth quickly when returned to an antibiotics-free liquid medium and formed colonies when spread on a solid medium. We developed a simple and highly efficient method of obtaining axenic microalgal cultures based on this observation. First, microalgal strains of different species or strains were treated with a mixture of ampicillin, gentamycin sulfate, kanamycin, neomycin and streptomycin (each at a concentration of 600 mg/L) for 3 days; they were then transferred to antibiotics-free medium for 5 days; and finally they were spread on solid f/2 media to allow algal colonies to form. With this method, five strains of Nannochloropsis sp. (Eustigmatophyceae), two strains of Cylindrotheca sp. (Bacillariophyceae), two strains of Tetraselmis sp. (Chlorodendrophyceae) and one strain of Amphikrikos sp. (Trebouxiophyceae) were purified successfully. The method shows promise for batch-purifying microalgal cultures.

  6. Study on Transformation of Oriental Hybrid Lily by Agrobacterium-mediated

    Institute of Scientific and Technical Information of China (English)

    FAN Jinping; ZHANG Chen; ZHANG Yingying; ZHOU Xiaojie; CHE Daidi

    2009-01-01

    An efficient procedure was described for the transformation of the monocotyledonous oriental hybrid lily, Lilium cv. Siberia, by Agrobacterium-mediatsd genetic transformation via leaves regeneration. The leaves of leaflets which derived from bulbs were sliced into 1.0 cm long and were co-cultivated with A. tumefaciens strain LBA4404/pB2AE12, which harbored a vector carrying the neomycin phosphotransferase, DREB2A genes in the T-DNA region. The suitable genetic transformation condition was determined as follows: the bacterial concentration reached 0.5-0.6 (OD600), 15 min infection time, 20 mg·L-1 acetosyingone, and 10.6 mmol·L-1 NH4NO3 medium was used for co-cultivation 3 days, delayed 7 days for selecting by 30 mg·L-1 kanamycin containing regeneration medium. Efficient shoot regeneration was observed on MS medium supplemented with 1.0 mg·L-1 naphthaleneacetic acid, 0.5 mg·L-1 benzyladenine and 0.1 mg·L-1 Kinetin after about 6 weeks culture. The presence of DREB1A gene in the genomic DNA of regenerated plants was detected by means of PCR analysis.

  7. Plasmid Mediated Antibiotic and Heavy Metal Resistance in Bacillus Strains Isolated From Soils in Rize, Turkey

    Directory of Open Access Journals (Sweden)

    Elif SEVİM

    2015-09-01

    Full Text Available Fifteen Bacillus strains which were isolated from soil samples were examined for resistance to 17 different antibiotics (ampicillin, methicillin, erythromycin, norfloxacin, cephalotine, gentamycin, ciprofloxacin, streptomycin, tobramycin, chloramphenicol, trimethoprim-sulfamethoxazole, tetracycline, vancomycin, oxacilin, neomycin, kanamycin and, novabiocin and to 10 different heavy metals (copper, lead, cobalt, chrome, iron, mercury, zinc, nickel, manganese and, cadmium and for the presence of plasmid DNA. A total of eleven strains (67% were resistant to at least one antibiotic. The most common resistance was observed against methicillin and oxacillin. The most resistance strains were found as Bacillus sp. B3 and Bacillus sp. B11. High heavy metal resistance against copper, chromium, zinc, iron and nickel was detected, but mercury and cobalt resistance was not detected, except for 3 strains (B3, B11, and B12 which showed mercury resistance. It has been determined that seven Bacillus strains have plasmids. The isolated plasmids were transformed into the Bacillus subtilis W168 and it was shown that heavy metal and antibiotic resistance determinants were carried on these plasmids. These results showed that there was a correlation between plasmid content and resistance for both antibiotic and heavy metal resistance

  8. Development of Transgenic Papaya through Agrobacterium-Mediated Transformation

    Directory of Open Access Journals (Sweden)

    Md. Abul Kalam Azad

    2013-01-01

    Full Text Available Transgenic papaya plants were regenerated from hypocotyls and immature zygotic embryo after cocultivation with Agrobacterium tumefaciens LBA-4404 carrying a binary plasmid vector system containing neomycin phosphotransferase (nptII gene as the selectable marker and β-glucuronidase (GUS as the reporter gene. The explants were co-cultivated with Agrobacterium tumefaciens on regeneration medium containing 500 mg/L carbenicillin + 200 mg/L cefotaxime for one week. The cocultivated explants were transferred into the final selection medium containing 500 mg/L carbenicillin + 200 mg/L cefotaxime + 50 mg/L kanamycin for callus induction as well as plant regeneration. The callus derived from the hypocotyls of Carica papaya cv. Shahi showed the highest positive GUS activities compared to Carica papaya cv. Ranchi. The transformed callus grew vigorously and formed embryos followed by transgenic plantlets successfully. The result of this study showed that the hypocotyls of C. papaya cv. Shahi and C. papaya cv. Ranchi are better explants for genetic transformation compared to immature embryos. The transformed C. papaya cv. Shahi also showed the maximum number of plant regeneration compared to that of C. papaya cv. Ranchi.

  9. A rapid and stable Agrobacterium-mediated transformation method of a medicinal plant Chelone glabra L.

    Science.gov (United States)

    Gao, Zhenrui; Li, Ying; Chen, Jinhua; Chen, Zhixing; Cui, Min-Long

    2015-03-01

    Transformation approach is a useful tool for the study of gene function, the mechanism of molecular regulation, and increase usefulness of components by reverse genetic approach in plants. In this study, we developed a stable and rapid method for Agrobacterium-mediated transformation of a medicinal plant Chelone glabra L. using leaf explants. Stable transformants were obtained using Agrobacterium tumefaciens strains GV2260 and GV3101 that harbored the binary vector pBI121 and contained the neomycin phosphotransferase gene (NPT II) as a selectable marker and a reporter gene β-glucuronidase (GUS). Putative transformants were identified by kanamycin selection and a histochemical assay. PCR and Southern blot analysis confirmed the integration of the GUS gene into transformed genomes as well as detected stable expression of the β-glucuronidase gene (GUS) by RT-PCR. Resulting transformed plants had morphologically normal phenotypes. This method requires two changes of medium and few leaf explants as well as the transformation efficiency of 2-8 % after 2-3 months of inoculation. This method can provide a quick and economical transformation method for reverse genetic approach to change the secondary metabolic pathway to increase useful components in C. glabra.

  10. A new high-frequency Agrobacterium-mediated transformation technique for Sesamum indicum L. using de-embryonated cotyledon as explant.

    Science.gov (United States)

    Chowdhury, Supriyo; Basu, Arpita; Kundu, Surekha

    2014-09-01

    In spite of the economic importance of sesame (Sesamum indicum L.) and the recent availability of its genome sequence, a high-frequency transformation protocol is still not available. The only two existing Agrobacterium-mediated transformation protocols that are available have poor transformation efficiencies of less than 2%. In the present study, we report a high-frequency, simple, and reproducible transformation protocol for sesame. Transformation was done using de-embryonated cotyledons via somatic embryogenic stages. All the critical parameters of transformation, like incubation period of explants in pre-regeneration medium prior to infection by Agrobacterium tumefaciens, cocultivation period, concentrations of acetosyringone in cocultivation medium, kanamycin concentration, and concentration of plant hormones, including 6-benzylaminopurine, have been optimized. This protocol is superior to the two existing protocols in its high regeneration and transformation efficiencies. The transformed sesame lines have been tested by PCR, RT-PCR for neomycin phosphotransferase II gene expression, and β-glucuronidase (GUS) assay. The regeneration frequency and transformation efficiency are 57.33 and 42.66%, respectively. T0 and T1 generation transgenic plants were analyzed, and several T1 plants homozygous for the transgenes were obtained.

  11. Drug: D01066 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available des 263 Suppurative dermatosis agents 2634 Antibiotics for external use D01066 Polymixin B sulfate (JP16); P...olymyxin B sulfate (USP/INN) 6 Agents against pathologic organisms and parasites 61 Antibiotics 612 Acting m... B sulfate (USP/INN) S SENSORY ORGANS S01 OPHTHALMOLOGICALS S01A ANTIINFECTIVES S01AA Antibiotics S01AA18 Po...L ANTIINFECTIVES A07AA Antibiotics A07AA05 Polymyxin B D01066 Polymixin B sulfate...ARY TRACT AND METABOLISM A07 ANTIDIARRHEALS, INTESTINAL ANTIINFLAMMATORY/ANTIINFECTIVE AGENTS A07A INTESTINA

  12. 卡那霉素对川芎愈伤组织诱导和生长的影响%Effects of Kanamycin on the Induction and Growth of Ligusticum chuanxiong Hort. Callus

    Institute of Scientific and Technical Information of China (English)

    蒋卫东; 唐琳; 杨苍劲; 马逾英; 蒋桂华; 陈放

    2007-01-01

    以川芎(Ligusticum chuanxiong Hort.)的根、茎、叶及其对应的愈伤组织为试验材料,研究不同浓度的卡那霉素对其诱导和生长的影响.不同的外植体对卡那霉素的敏感度不同,根的敏感度高于茎,茎的敏感度高于叶片.方差分析和总体观察的结果表明,在川芎的遗传转化中,卡那霉素的最佳抗性筛选浓度范围为20~40 mg/L.

  13. Identification of Kanamycin-Resistant Enterococci in 92 Undergraduates%92例大学生耐卡那霉素肠球菌的分离鉴定及其相关因素

    Institute of Scientific and Technical Information of China (English)

    黄静丽; 姜智晖; 曾香玲; 吴健仪; 李文华; 麦璟莹; 苏丹红; 李孜

    2008-01-01

    目的 分离鉴定健康大学生粪便中的耐卡那霉素肠球菌,并调查相关因素.方法 用卡那霉素ELB选择培养基分离培养92份健康大学生粪便中的肠球菌,调查他们的健康状况及生活习惯等.结果 共鉴定出21例携带卡那霉素抗性肠球菌,71例阴性.耐卡那霉素肠球菌的分离率与饮食辛辣的相关性具有统计学意义(X2=5.75,P<0.05);而与消化不良、胃肠不适和便秘的相关性无统计学意义(X2<3.84,P>0.05).结论 耐卡那霉素肠球菌的分离率与饮食辛辣相关,与消化不良、胃肠不适和便秘等无相关性,其机制及其临床意义有待进一步的研究和随访.

  14. 定性PCR技术及卡那霉素对8个番茄品种的转基因检测%GMO Detection in Eight Different Tomato Varieties by Using Qualitative PCR Technique and Kanamycin-Resistance

    Institute of Scientific and Technical Information of China (English)

    许爽; 褚云霞; 张辉; 朱龙英; 朱为民

    2009-01-01

    本研究利用定性PCR技术对8个不同番茄品种中外源基因35S启动子、NPTⅡ基因和NOS终止子进行检测.实验结果表明,阳性对照可以稳定扩增到预期的大小片段,而待检测品种和阴性对照则没有扩增到预期产物.利用100 mg/L的卡那霉素对8个不同番茄品种的种子进行萌发试验,再用50 mg/L的卡那霉素对番茄外植体(子叶和下胚轴)进行抗性验证,结果显示卡那霉素抗性筛选实验结果与定性PCR结果一致.本文通过对这两种检测方法的优缺点进行研究分析及比较,初步建立了一个对不同品种番茄进行转基因检测的体系.

  15. 葛根素对豚鼠卡那霉素耳中毒听觉诱发电位的影响%The Effects of Puerarin on Brainstem Auditory Evoked Potentials of Ototoxicity with Kanamycin in Guinea-pigs

    Institute of Scientific and Technical Information of China (English)

    金成文; 王玉良; 陆洪英; 程秀臻; 刘儒林

    2002-01-01

    观察葛根素在豚鼠卡那霉素耳中毒中的作用.将豚鼠30只随机分两组:对照组15只,肌注卡那霉素400mg/kg/d,共12天;葛根素组15只,除肌注卡那霉素400mg/kg/d外,腹腔注射葛根素10mg/kg/d,共12天.以听觉诱发电位为指标进行组间比较,听觉诱发电位阈值对照组明显高于葛根素组(P<0.05);对照组诱发电位各波幅值则明显低于葛根素组(P<0.01).葛根素能减轻卡那霉素耳毒性,对卡那霉素耳中毒具有一定的保护作用,为耳毒性抗生素致聋的防治提供了一条新的研究途径.

  16. Sequences of two related multiple antibiotic resistance virulence plasmids sharing a unique IS26-related molecular signature isolated from different Escherichia coli pathotypes from different hosts.

    Directory of Open Access Journals (Sweden)

    Carola Venturini

    Full Text Available Enterohemorrhagic Escherichia coli (EHEC and atypical enteropathogenic E. coli (aEPEC are important zoonotic pathogens that increasingly are becoming resistant to multiple antibiotics. Here we describe two plasmids, pO26-CRL125 (125 kb from a human O26:H- EHEC, and pO111-CRL115 (115kb from a bovine O111 aEPEC, that impart resistance to ampicillin, kanamycin, neomycin, streptomycin, sulfathiazole, trimethoprim and tetracycline and both contain atypical class 1 integrons with an identical IS26-mediated deletion in their 3´-conserved segment. Complete sequence analysis showed that pO26-CRL125 and pO111-CRL115 are essentially identical except for a 9.7 kb fragment, present in the backbone of pO26-CRL125 but absent in pO111-CRL115, and several indels. The 9.7 kb fragment encodes IncI-associated genes involved in plasmid stability during conjugation, a putative transposase gene and three imperfect repeats. Contiguous sequence identical to regions within these pO26-CRL125 imperfect repeats was identified in pO111-CRL115 precisely where the 9.7 kb fragment is missing, suggesting it may be mobile. Sequences shared between the plasmids include a complete IncZ replicon, a unique toxin/antitoxin system, IncI stability and maintenance genes, a novel putative serine protease autotransporter, and an IncI1 transfer system including a unique shufflon. Both plasmids carry a derivate Tn21 transposon with an atypical class 1 integron comprising a dfrA5 gene cassette encoding resistance to trimethoprim, and 24 bp of the 3´-conserved segment followed by Tn6026, which encodes resistance to ampicillin, kanymycin, neomycin, streptomycin and sulfathiazole. The Tn21-derivative transposon is linked to a truncated Tn1721, encoding resistance to tetracycline, via a region containing the IncP-1α oriV. Absence of the 5 bp direct repeats flanking Tn3-family transposons, indicates that homologous recombination events played a key role in the formation of this complex

  17. Assessing specific oligonucleotides and small molecule antibiotics for the ability to inhibit the CRD-BP-CD44 RNA interaction.

    Directory of Open Access Journals (Sweden)

    Dustin T King

    Full Text Available Studies on Coding Region Determinant-Binding Protein (CRD-BP and its orthologs have confirmed their functional role in mRNA stability and localization. CRD-BP is present in extremely low levels in normal adult tissues, but it is over-expressed in many types of aggressive human cancers and in neonatal tissues. Although the exact role of CRD-BP in tumour progression is unclear, cumulative evidence suggests that its ability to physically associate with target mRNAs is an important criterion for its oncogenic role. CRD-BP has high affinity for the 3'UTR of the oncogenic CD44 mRNA and depletion of CRD-BP in cells led to destabilization of CD44 mRNA, decreased CD44 expression, reduced adhesion and disruption of invadopodia formation. Here, we further characterize the CRD-BP-CD44 RNA interaction and assess specific antisense oligonucleotides and small molecule antibiotics for their ability to inhibit the CRD-BP-CD44 RNA interaction. CRD-BP has a high affinity for binding to CD44 RNA nts 2862-3055 with a Kd of 645 nM. Out of ten antisense oligonucleotides spanning nts 2862-3055, only three antisense oligonucleotides (DD4, DD7 and DD10 were effective in competing with CRD-BP for binding to 32P-labeled CD44 RNA. The potency of DD4, DD7 and DD10 in inhibiting the CRD-BP-CD44 RNA interaction in vitro correlated with their ability to specifically reduce the steady-state level of CD44 mRNA in cells. The aminoglycoside antibiotics neomycin, paramomycin, kanamycin and streptomycin effectively inhibited the CRD-BP-CD44 RNA interaction in vitro. Assessing the potential inhibitory effect of aminoglycoside antibiotics including neomycin on the CRD-BP-CD44 mRNA interaction in cells proved difficult, likely due to their propensity to non-specifically bind nucleic acids. Our results have important implications for future studies in finding small molecules and nucleic acid-based inhibitors that interfere with protein-RNA interactions.

  18. 缘管浒苔游孢子转化系统选择标记的研究%Studies on the Sensitivity of Ulva Linza Spores to Antibiotics

    Institute of Scientific and Technical Information of China (English)

    卢永忠; 杨中飞

    2014-01-01

    选择标记的确定是基因工程研究的一个重要环节。主要研究了缘管浒苔游孢子对青霉素、氯霉素等9种抗生素的敏感性,并与不同发育时期的藻体进行了比较。结果表明:缘管浒苔游孢子对氯霉素、新霉素、红霉素、G-418敏感性较高,在10μg/mL时均有明显的致死效应;不同发育时期浒苔的抗生素敏感性不同,成熟度越高,敏感性越低;对青霉素、卡那霉素、四环素、潮霉素、链霉素不敏感。本实验结果提示氯霉素、红霉素、G-418、新霉素可作为缘管浒苔游孢子转基因研究理想的选择试剂。%To determine the proper screening marker is a key step for genetic engineering. In this study the sensitivity of Ulva linza spores to nine kinds of antibiotics such as penicillin,chloramphenicol and so on was investigated,and it was also compared with Ulva linza in different development period. The results showed that Ulva linza spores were more sensitive to chloramphenicol,erythromycin,neomycin and G -418,all exhibiting obvious killing effects under the concentration of 10μg/mL,whereas mature alga was less sensitive;they were not sensitive to penicillin, kanamycin, homomycin, streptomycin and tetracycline. This indicated that chloramphenicol,erythromycin,neomycin and G-418 could be used as screening agents for gene transferring of Ulva linza spores.

  19. Antimicrobial sensitivity profile of Staphylococcus spp. Isolated from clinical mastitis

    Directory of Open Access Journals (Sweden)

    Thamires Martins

    2012-12-01

    Full Text Available Inflammation of the mammary gland, which is also known as mastitis, occupies a prominent place among the diseases that affect dairy cattle, having a great economic importance in the dairy sector. Mastitis may have different origins, however, infectious mastitis is the most frequent and represents a risk to public health due to the propagation of microorganisms through milk. Staphylococcus spp. are considered the microorganisms that cause the greatest losses in milk production, being that Staphylococcus aureus is the pathogen of major importance because they present high resistence to antimicrobials. Empirical treatment, without prior identification of the pathogens and their resistance profile, may contribute to the emergence of multidrug-resistant strains and risk the efficiency of the antimicrobial. In that scenery, the study aimed to evaluate the resistance profile of Staphylococcus spp. against some antimicrobials used in the treatment of cows with clinical mastitis. The study was conducted on a property in the state of São Paulo from January 2011 to June 2012. We evaluated 29 lactating cows that present clinical mastitis in, at least, one mammary quarter. The diagnosis of clinical mastitis was performed by evaluating the clinical signs and also by Tamis test. Samples of milk from mammary quarters were collected aseptically in sterile tubes for microbiological evaluation. Microorganisms were isolated on sheep blood agar 5% and Sabouraud agar with chloramphenicol. The sensitivity profile of Staphylococcus spp. to the antibiotics ampicillin, cephalexin, ceftiofur, cefaclor, gentamicin, kanamycin, neomycin, penicillin G and oxacillin, was tested by disk diffusion test on Mueller-Hinton agar. From a total of 106 samples of milk analyzed, 64 (60.38% presented microbiological growth, being observed isolation of Streptococcus spp. 29 (34.52%, Staphylococcus spp. 28 (33.33%, Corynebacterium spp. 17 (20.24%, filamentous fungi 4 (4.76%, yeast 4 (4

  20. African Journal of Biotechnology - Vol 9, No 49 (2010)

    African Journals Online (AJOL)

    Factors impacting on the microbiological quality and safety of processed hake ... Microbial transformation of neomycin by a mutant of neomycin-producing ... Antioxidant and antibacterial activities of ethanolic extracts of Asparagus officinalis cv.

  1. Drug: D02139 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available D02139 Mixture, Drug Hydrocortisone acetate - neomycin sulfate - colistin sulfate -...3 Hydrocortisone and antiinfectives D02139 Hydrocortisone acetate - neomycin sulfate - colistin sulfate - thonzonium bromide mixt PubChem: 7849200 ...

  2. Domain Modeling: NP_061870.1 [SAHG[Archive

    Lifescience Database Archive (English)

    Full Text Available NP_061870.1 chr1 Crystal structure of the bacterial ribosome from Escherichia coli in complex with neomycin.... This file contains the 30S subunit of the first 70S ribosome, with neomycin bound.

  3. Drug: D05142 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available MMATORY/ANTIINFECTIVE AGENTS A07A INTESTINAL ANTIINFECTIVES A07AA Antibiotics A07... RESPIRATORY SYSTEM R02 THROAT PREPARATIONS R02A THROAT PREPARATIONS R02AB Antibiotics... R02AB01 Neomycin D05142 Neomycin undecylenate (USAN) S SENSORY ORGANS S01 OPHTHALMOLOGICALS S01A ANTIINFECTIVES S01AA Antibioti...cs S01AA03 Neomycin D05142 Neomycin undecylenate (USAN) S02 OTOLOGICALS S02A ANTIIN

  4. 多重耐药鲍曼不动杆菌对氨基糖苷类药物耐药性及其耐药基因(ant)的研究%Study on aminoglycosides resistance and its resistance genes in multidrug resistant Acinetobacter baumannii

    Institute of Scientific and Technical Information of China (English)

    顾建文; 姚丽娜; 史伟峰

    2008-01-01

    Objective To investigate the resistance of multi-drug resistant Acinetobacter baumannii against aminoglycosides and its resistance genes.Methods The minimal inhibitory concentrations (MICs) of seven antimicrobials (gentamicin, amikacin, tobramycine, netilmicin, neomycin, streptomycin and kanamycin) against 20 strains of multi-drug resistant Acinetobacter baumannii were detected by agar dilution method. Meanwhile, two resistance genes of aminoglycoside nucleotidyltransferase were amplified by PCR and vertified by DNA sequencer.Results It was found that MIC50 and MIC90 of gentamicin, amikacin, streptomycin and kanamycin in 20 strains of Acinetobacter baumannii were all above 1 024 mg/L, and their resistant rates were 95%, 95%, 100% and 90% respectively, while the resistant rates of tobramycin, netilmicin and neomycin were 85%, 90% and 40% respectively. Two aminoglycoside-modification genes ant(2″)-Ⅰ and ant(3″)-Ⅰ were detected in 20 strains, with 55% and 80% of positive rate respectively. The double positive rate of two resistance genes was 50%.Conclusion The resistance of multi-drug resistant Acinetobacter baumannii against aminoglycosides was closely associated with ant(2″)-Ⅰ and ant(3″)-Ⅰ genes.%目的 了解多重耐药鲍曼不动杆菌对氨基糖苷类药物修饰酶的耐药性及其耐药基因.方法 用琼脂稀释法检测庆大霉素、阿米卡星、链霉素、卡那霉素、妥布霉素、奈替米星、新霉素7种药物对20株多重耐药性鲍曼不动杆菌的最低抑菌浓度(MIC);用PCR法检测两种氨基糖苷类药物核苷转移酶基因,并使用DNA测序加以证实.结果 庆大霉素、阿米卡星、链霉素、卡那霉素对20株鲍曼不动杆菌的MIC50和MIC90均大于1 024 mg/L,耐药率分别为95%、95%、100%和90%;而妥布霉素、奈替米星和新霉素的耐药率分别为85%、90%、40%.从20株菌中检出ant(2″)-Ⅰ、ant(3″)-Ⅰ两种修饰酶基因,阳性率分别为55%、80%,10

  5. [Investigation of pathogenic phenotypes and virulence determinants of food-borne Salmonella enterica strains in Caenorhabditis elegans animal model].

    Science.gov (United States)

    Aksoy, Deniz; Şen, Ece

    2015-10-01

    Salmonellosis, caused by non-typhoidal Salmonella enterica serovars with the consumption of contaminated food, is one of the leading food-borne disease that makes microbial food safety an important public health issue. This study was performed in order to determine the antibiotic resistance, serotyping, plasmid profiles and pathogenicity potentials of food-borne Salmonella isolates in Caenorhabditis elegans animal model system in Edirne province, located at Thrace region of Turkey. In this study, 32 Salmonella isolates, of which 26 belonged to Infantis, four to Enteritidis, one to Telaviv and one to Kentucky serovars, isolated from chicken carcasses were used. Antibiotic resistance profiles were determined by disc diffusion and broth microdilution methods. A new C.elegans nematode animal model system was used to determine the pathogenicity potential of the isolates. The antibiotic resistance profiles revealed that one (3.1%) isolate was resistant to gentamicin, two (6.2%) to ciprofloxacin, three (9.4%) to ampicillin, 18 (56.3%) to kanamycin, 19 (60.8%) to neomycin, 25 (78.1%) to tetracycline, 25 (78.1%) to trimethoprim, 26 (81.25%) to nalidixic acid, 27 (84.4%) to streptomycin and 32 (100%) to sulfonamide. All of the 32 strains were susceptible to chloramphenicol and ampicillin/sulbactam. High levels of resistance to streptomycin, nalidixic acid, tetracycline, trimethoprim, sulfonamide, kanamycin and neomycin was determined. According to the plasmid analysis, six isolates (18.75%) harboured 1-3 plasmids with sizes between 1.2 and 42.4 kb. In C.elegans nematode animal model system, the time (in days) required to kill 50% (TD50) of nematodes was calculated for each experimental group. TD50 values of the nematode group fed with S.Typhimurium ATCC 14028 that was used as the positive control and another group fed with E.coli OP50 as the negative control were 4.2 ± 0.5 days and 8.0 ± 0.02 days, respectively. TD50 of the groups fed with Salmonella isolates ranged

  6. Antimicrobial susceptibility of Campylobacter sp strains isolated from calves with and without diarrhea in Minas Gerais state, Brazil Susceptibilidade a antimicrobianos de amostras de Campylobacter sp isoladas de bezerros com e sem diarréia, no estado de Minas Gerais, Brasil

    Directory of Open Access Journals (Sweden)

    Karina Leite Miranda

    2007-06-01

    Full Text Available The antimicrobial susceptibility of 25 Campylobacter sp strains isolated from calves with and without diarrhea - 7 C. coli, 16 C. fetus and 2 C. jejuni was studied by the disk diffusion method. Eleven antimicrobial agents were tested amikacin, ampicillin, kanamycin, chloramphenicol, erythromycin, gentamicin, neomycin, nitrofurantoin, penicillin G, tetracycline and sulfamethoxazole-trimethoprim. All Campylobacter sp strains were susceptible to amikacin, ampicillin, chloramphenicol, erythromycin, gentamicin, neomycin and nitrofurantoin. Three strains were moderately susceptible to kanamycin (2 C. coli and 1 C. fetus. All the strains were resistant to penicillin G. Two C. fetus strains were moderately susceptible to sulfamethoxazole-trimethoprim and 1 C. coli, 9 C. fetus and 2 C. jejuni strains were resistant. Two C. fetus strains were moderately susceptible to tetracycline and 3 C. coli, 2 C. fetus and 1 C. jejuni strains were resistant. Eleven strains showed multidrug resistance (2 C. coli, 8 C. fetus and 1 C. jejuni. There was no correlation between resistance of Campylobacter sp strains to antimicrobials and the occurrence of diarrhea in calves. The frequency of resistance and, most importantly, multi drug resistance found among Campylobacter sp strains isolated from calves in Minas Gerais, Brazil, were high and the patterns of resistance observed are related to the antimicrobials agents most largely used in cattle in Brazil.Foi estudado o perfil de susceptibilidade aos antimicrobianos de 25 amostras de Campylobacter sp isoladas de bezerros com e sem diarréia (7 C. coli, 16 C. fetus e 2 C. jejuni. Foram testados pelo método de difusão 11 agentes antimicrobianos: amicacina, ampicilina, canamicina, cloranfenicol, eritromicina, gentamicina, neomicina, nitrofurantoína, penicilina G, tetraciclina e sulfametoxazole-trimetoprim. Todas as amostras de Campylobacter sp foram susceptíveis a amicacina, ampicilina, cloranfenicol, eritromicina

  7. The expression of the Saccharomyces cerevisiae HAL1 gene increases salt tolerance in transgenic watermelon [Citrullus lanatus (Thunb.) Matsun. & Nakai.].

    Science.gov (United States)

    Ellul, P; Ríos, G; Atarés, A; Roig, L A; Serrano, R; Moreno, V

    2003-08-01

    An optimised Agrobacterium-mediated gene transfer protocol was developed in order to obtain watermelon transgenic plants [Citrullus lanatus (Thunb.) Matsun. & Nakai.]. Transformation efficiencies ranged from 2.8% to 5.3%, depending on the cultivar. The method was applied to obtain genetically engineered watermelon plants expressing the Saccharomyces cerevisiae HAL1 gene related to salt tolerance. In order to enhance its constitutive expression in plants, the HAL1 gene was cloned in a pBiN19 plasmid under control of the 35S promoter with a double enhancer sequence from the cauliflower mosaic virus and the RNA4 leader sequence of the alfalfa mosaic virus. This vector was introduced into Agrobacterium tumefaciens strain LBA4404 for further inoculation of watermelon half-cotyledon explants. The introduction of both the neomycin phosphotransferase II and HAL1 genes was assessed in primary transformants (TG1) by polymerase chain reaction analysis and Southern hybridisation. The expression of the HAL1 gene was determined by Northern analysis, and the diploid level of transgenic plants was confirmed by flow cytometry. The presence of the selectable marker gene in the expected Mendelian ratios was demonstrated in TG2 progenies. The TG2 kanamycin-resistant plantlets elongated better and produced new roots and leaves in culture media supplemented with NaCl compared with the control. Salt tolerance was confirmed in a semi-hydroponic system (EC=6 dS m(-1)) on the basis of the higher growth performance of homozygous TG3 lines with respect to their respective azygous control lines without the transgene. The halotolerance observed confirmed the inheritance of the trait and supports the potential usefulness of the HAL1 gene of S. cerevisiae as a molecular tool for genetic engineering of salt-stress protection in other crop species.

  8. Antibiotic Sensitivity Pattern of Staphylococcus aureus and Escherichia coli Isolated from Bovine Fresh Milk (POLA SENSITIVITAS ANTIBIOTIK TERHADAP STAPHYLOCOCCUS AUREUS DAN ESCHERICHIA COLI YANG DIISOLASI DARI SUSU SAPI SEGAR

    Directory of Open Access Journals (Sweden)

    Lucia Ratna Winata Muslimin

    2016-01-01

    Full Text Available The aims of this study were to determine the sensitivity of S.aureus and E. coli isolated fromfresh milk against against several antibiotics and to determine the safety of the milk for humancomsumsion. Milk was collected from milking diary cow and was used for the bacterial isolation. E.coli were were identified using Total Plate Count (TPC, Gram staining, their growth on Endo Agarand Eosin MethyleneBlue Agar, Biochemical analysis including glucose, lactose, sucrose,maltose, andsorbitol would be followed by Sorbitol Mac Conkey Agar Test for the identification of E.coliO157:H7.The isolation and identification of S.aureus were performed using Gram stain, TPC, growth on BairdParker Agar and Mannitol Salt Agar. S. aureus and S. epidermidis were differentiated by coagulaseand catalase tests. The antibiotic sensitivity tests for both S. aureus and E.coli were carried out usingthe following antibiotics: ampicillin, bacitracin, vancomycin, cefoperazone, ceftriaxone, cefotamine,cefuroxime, cefepime, cefazoline, ceftazidime, chloramphenicol, tetracycline, doxycycline, amikacin,kanamycin, neomycin, ertapenem, meropenem, imipenem, erythromycin, gentamycin, nalidixic acid,ciprofloxacin, levofloxacine, norfloxacine, ofloxacin, and novobiocin. Fresh milk obtained from thefarm was positive for S.aureus and E.coli and resistant to most antibiotics tested. The best antibioticsfor S. aureus were imipenem (54.1 mm, ampicillin (42.3 mm, cefazolin (41.6 mm, doxycycline (41.15mm, and for E.coli were Imipenem (30.1 mm, ertapenem (29.5 mm, and meropenem (25.35 mm. Thebovine fresh milk examined was contaminated by S.aureus and E.coli and to some extent, were alsoresistant to most antibiotics tested.

  9. Study of antimicrobial resistance and physiological biomarkers with special reference to Salmonellosis in diarrheic foals in Punjab, Pakistan.

    Science.gov (United States)

    Haq, Ikramul; Durrani, Aneela Zameer; Khan, Muhammad Sarwar; Mushtaq, Muhammad Hassan; Ahmad, Imtiaz

    2017-08-04

    Antimicrobial resistance results in selective colonization in animals. In the present study, 447 diarrheic foals (235 horse foals, 165 donkey foals and 47 mule foal) were selected from Lahore and Sahiwal districts of Punjab, Pakistan. Fresh fecal and blood samples from diarrheic foals were collected for isolation and confirmation of Salmonella Polymerase chain reaction. Results revealed that 50 (11.25%) foals (horse n=29, donkey n=12 and mule n=9) were positive. Fifty Salmonella enterica isolates belonging to 7 serovars S. Paratyphi B (15), S. Saintpaul (7), S. Newport (6), S. Typhimu-rium (11), S. Kottbus (4), S. Lagos (2), and S. enterica ssp salamae (5). Salmonella was common in foals that visited veterinary hospital, as compared to those in stud farms and individual foals reared in low income household. Out of the total 50 samples, 92% of isolates were resistant to three or more than three antimicrobials. The highest resistance (86%) was against Sulphamethoxazole (23.75mg) and lowest (4%) against trimethoprime (5mg). The isolates also showed resistance against Doxycycline (30mg), Oxytetracycline (30mg), Streptomycin (10mg), Neomycin (30mg), Amikacin (30mg), chloramphenicol (30mg), Ampicillin (10mg), Amoxicillin (10mg), kanamycin (30mg), Norfloxacin (10mg), Gentamicin 10mg, Cefotaxime (30mg), Ciprofloxacin (5mg) and Ceftriaxone (30mg). Blood analysis of salmonella infected foals showed That Hemoglobin, PCV and TEC were significantly higher and (while) TLC, PCV, Monocytes, Lymphocytes, Basophils, Eosinophil and Neutrophils were significantly lower than normal. Albumin were lower and BNU, Biluribin, ALT and creatinine were higher than normal values. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Heat-shock-mediated elimination of the nptII marker gene in transgenic apple (Malus×domestica Borkh.).

    Science.gov (United States)

    Herzog, Katja; Flachowsky, Henryk; Deising, Holger B; Hanke, Magda-Viola

    2012-04-25

    Production of marker-free genetically modified (GM) plants is one of the major challenges of molecular fruit breeding. Employing clean vector technologies, allowing the removal of undesired DNA sequences from GM plants, this goal can be achieved. The present study describes the establishment of a clean vector system in apple Malus×domestica Borkh., which is based on the use of the neomycin phosphotransferase II gene (nptII) as selectable marker gene and kanamycin/paramomycin as selective agent. The nptII gene can be removed after selection of GM shoots via site-specific excision mediated by heat-shock-inducible expression of the budding yeast FLP recombinase driven by the soybean Gmhsp17.5-E promoter. We created a monitoring vector containing the nptII and the flp gene as a box flanked by two direct repeats of the flp recognition target (FRT) sites. The FRT-flanked box separates the gusA reporter gene from the Cauliflower Mosaic Virus 35S (CaMV 35S) promoter. Consequently, GUS expression does only occur after elimination of the FRT-flanked box. Transformation experiments using the monitoring vector resulted in a total of nine transgenic lines. These lines were investigated for transgenicity by PCR, RT-PCR and Southern hybridization. Among different temperature regimes tested, exposure to 42 °C for 3.5 to 4h led to efficient induction of FLP-mediated recombination and removal of the nptII marker gene. A second round of shoot regeneration from leaf explants led to GM apple plants completely free of the nptII gene.

  11. Agrobacterium tumefaciens-mediated transformation of blueberry (Vaccinium corymbosum L.).

    Science.gov (United States)

    Song, Guo-Qing; Sink, K C

    2004-12-01

    Transient expression studies using blueberry leaf explants and monitored by beta-glucuronidase (GUS) assays indicated Agrobacterium tumefaciens strain EHA105 was more effective than LBA4404 or GV3101; and the use of acetosyringone (AS) at 100 microM for inoculation and 6 days co-cultivation was optimum compared to 2, 4, 8, 10 or 12 days. Subsequently, explants of the cultivars Aurora, Bluecrop, Brigitta, and Legacy were inoculated with strain EHA105 containing the binary vector pBISN1 with the neomycin phosphotransferase gene (nptII) and an intron-interrupted GUS gene directed by the chimeric super promoter (Aocs)3AmasPmas. Co-cultivation was for 6 days on modified woody plant medium (WPM) plus 100 microM AS. Explants were then placed on modified WPM supplemented with 1.0 mg l(-1) thidiazuron, 0.5 mg l(-1) alpha-naphthaleneacetic, 10 mg l(-1) kanamycin (Km), and 250 mg l(-1) cefotaxime. Selection for Km-resistant shoots was carried out in the dark for 2 weeks followed by culture in the light at 30 microE m(-2) s(-1) at 25 degrees C. After 12 weeks, selected shoots that were both Km resistant and GUS positive were obtained from 15.3% of the inoculated leaf explants of cultivar Aurora. Sixty-eight independent clones derived from such shoots all tested positive by the polymerase chain reaction using a nptII primer. Eight of eight among these 68 clones tested positive by Southern hybridization using a gusA gene derived probe. The transformation protocol also yielded Km-resistant, GUS-positive shoots that were also PCR positive at frequencies of 5.0% for Bluecrop, 10.0% for Brigitta and 5.6% for Legacy.

  12. Instability of multiple drug resistance plasmids in Salmonella typhimurium isolated from poultry.

    Science.gov (United States)

    Brown, D J; Threlfall, E J; Rowe, B

    1991-04-01

    Plasmids in five strains of Salmonella typhimurium resistant to ampicillin, chloramphenicol, gentamicin, neomycin/kanamycin, streptomycin, sulphonamides, tetracyclines and trimethoprim (ACGKSSuTTm), CGKSSuTTm, ACSSuT or CSSuT which had been isolated from poultry in the first 3 months of 1989 have been characterized and compared with plasmids in two strains of R-types ACGKSSuTTm and ASSuTTm isolated from two patients later in the year. With the exception of the human isolate of R-type ASSuTTm, all strains carried two non-conjugative plasmids, one coding for SSu and belonging to incompatibility group Q, and a second coding for multiple resistance and belonging to the FIme incompatibility group. The human isolate of R-type ASSuTTm did not carry the IncQ SSu plasmid but like the poultry isolates, carried a non-conjugative FIme plasmid. Restriction endonuclease digestion with the enzymes EcoR I, Pst I and Hind III demonstrated that the FIme plasmids from strains of different R-types showed a high degree of homology but exhibited numerous fragment size polymorphisms. The restriction digest fingerprint of plasmids in the human isolate of R-type ACGKSSuTTm was indistinguishable from a poultry isolate of the same R-type. Analysis of segregants of one of the poultry isolates of R-type ACGKSSuTTm demonstrated that resistance determinants could be rapidly lost from the FIme plasmid to give rise to a number of R-types and fingerprint patterns. Loss of tetracycline resistance from this plasmid appeared to be correlated with the integration of other plasmid-mediated resistances into the bacterial chromosome. Evidence is presented for the rapid loss of antimicrobial resistance determinants from a multiple resistance plasmid of the FIme incompatibility group in response to withdrawal of antibiotic selective pressure.

  13. A novel multidrug resistance plasmid isolated from an Escherichia coli strain resistant to aminoglycosides.

    Science.gov (United States)

    Sun, Hui; Li, Shasha; Xie, Zhijing; Yang, Fangfang; Sun, Yani; Zhu, Yanli; Zhao, Xiaomin; Jiang, Shijin

    2012-07-01

    Previous studies have reported several different plasmids that confer multidrug resistance (MDR) including resistance to aminoglycosides. In this study, we investigated the aminoglycoside resistance patterns for 224 Escherichia coli isolates from diseased chickens and ducks in China, characterized a novel MDR plasmid, and collected prevalence data on similar resistance plasmids. Antibiotic susceptibilities were determined using disc diffusion and the microdilution method. The plasmid pXZ was analysed by restriction fragment length polymorphism (RFLP) with EcoRI and SalI, and sequenced. The prevalence of similar resistance plasmids was assessed by multiplex PCR and by RFLP analysis. Among the 224 E. coli isolates, 189 (84.4%) were resistant to streptomycin, 125 (55.8%) were resistant to kanamycin, 116 (51.8%) were resistant to gentamicin, 106 (47.3%) were resistant to neomycin and 98 (43.8%) were resistant to amikacin. Among the 224 E. coli isolates, 17 contained a plasmid with the MDR-encoding region of pXZ, which showed high-level resistance to aminoglycosides (MICs of gentamicin and amikacin ≥ 512 mg/L). The plasmid pXZ was digested into five fragments by EcoRI and six fragments by SalI. The plasmid pXZ was a circular DNA molecule of 76635 bp with a 51.65% guanine + cytosine content and included four resistance genes (rmtB, fosA3, bla(TEM-1) and bla(CTX-M-24)). A novel MDR plasmid, pXZ, harbouring four resistance genes (rmtB, fosA3, bla(TEM-1) and bla(CTX-M)) was identified. To our knowledge, this is the first report of an aminoglycoside resistance plasmid harbouring the fosA3 gene.

  14. Antimicrobial resistance trends among Salmonella isolates obtained from dairy cattle in the northeastern United States, 2004-2011.

    Science.gov (United States)

    Cummings, Kevin J; Perkins, Gillian A; Khatibzadeh, Sarah M; Warnick, Lorin D; Altier, Craig

    2013-04-01

    Monitoring antimicrobial resistance trends among bacteria isolated from food animals and people is necessary to inform public policy regarding appropriate antimicrobial use. Our objectives were to describe the antimicrobial resistance status of Salmonella isolates from dairy cattle in the northeastern United States and to identify trends in resistance to various antimicrobial agents over time. Data were collected retrospectively for all bovine Salmonella isolates that were obtained from samples submitted to Cornell University's Animal Health Diagnostic Center between January 1, 2004 and December 31, 2011. Temporal trends in the prevalence of resistant Salmonella were investigated for each antimicrobial agent using the Cochran-Armitage trend test. Antimicrobial susceptibility testing was performed on 2745 bovine Salmonella isolates from clinical samples submitted during the study period. Overall resistance to each antimicrobial agent ranged from 0% (amikacin, ciprofloxacin, and nalidixic acid) to 72.0% (sulfadimethoxine). There was evidence of a significantly decreasing trend in prevalence of resistance to most agents: amoxicillin/clavulanic acid (AUG), ampicillin (AMP), cefoxitin (FOX), ceftiofur (TIO), ceftriaxone (AXO), chloramphenicol (CHL), chlortetracycline (CTET), florfenicol (FFN), kanamycin (KAN), neomycin (NEO), oxytetracycline (OXY), spectinomycin (SPE), streptomycin (STR), sulfadimethoxine (SDM), sulfisoxazole (FIS), and tetracycline (TET). Among the 265 isolates that were tested using the National Antimicrobial Resistance Monitoring System (NARMS) panel, the most common resistance patterns were pansusceptible (54.0%), AUG-AMP-FOX-TIO-AXO-CHL-KAN-STR-FIS-TET (18.1%), and AUG-AMP-FOX-TIO-AXO-CHL-STR-FIS-TET (12.1%). Increasing prevalence of S. enterica serovar Cerro over the course of the study period presumably had an impact on the observed resistance trends. Nevertheless, these results do not support the notion that the current level of antimicrobial

  15. Characterization and identification of streptococci from golden pompano in China.

    Science.gov (United States)

    Cai, X H; Peng, Y H; Wang, Z C; Huang, T; Xiong, X Y; Huang, Y C; Wang, B; Xu, L W; Wu, Z H

    2016-05-26

    Streptococcal infections cause significant mortality and high economic losses in the fish farm industry worldwide, including in the culture of golden pompano Trachinotus ovatus L., a species gaining popularity in China. A total of 9 streptococcal strains were isolated from cage-cultured diseased golden pompano in Beihai, Zhanjing, and Shenzhen, China, between 2012 and 2014. Conventional and rapid identification systems were used to determine that the isolates were Streptococcus agalactiae, S. iniae, and S. dysgalactiae subsp. dysgalactiae. All isolates were gram-positive cocci cells in pairs or short-chain, non-motile, catalase negative, α or β hemolytic cocci. The results of multiplex PCR assays and 16S rRNA BLAST analysis also showed that the β hemolytic strains were S. agalactiae and S. iniae and the α hemolytic strain was S. dysgalactiae subsp. dysgalactiae, respectively. Pathogenicity assays revealed that S. agalactiae (lethal dose [LD50]: 6.38 × 10(4) CFU ml(-1)) was more virulent for golden pompano than S. iniae (LD50: 1.47 × 10(7) CFU ml(-1)) and S. dysgalactiae subsp. dysgalactiae (LD50: 2.57 × 10(6) CFU ml(-1)) when they were challenged by intraperiotoneal (i.p.) injection. The results of antibiotic susceptibility showed that all strains were extremely susceptible to cefradine, erythromycin, and cefotaxime but resistant to gentamicin, penicillin G, novobiocin, neomycin, ciprofloxacin, roxithromycin, furazolidone, enrofloxacin, norfloxacin, kanamycin, ampicillin, tetracycline, and vancomycin This is the first report of a phenomenon of golden pompano coinfection with S. agalactiae and S. iniae, which will contribute to the diagnosis and prevention of streptococcicosis.

  16. Complete sequence of a plasmid from a bovine methicillin-resistant Staphylococcus aureus harbouring a novel ica-like gene cluster in addition to antimicrobial and heavy metal resistance genes.

    Science.gov (United States)

    Feßler, Andrea T; Zhao, Qin; Schoenfelder, Sonja; Kadlec, Kristina; Brenner Michael, Geovana; Wang, Yang; Ziebuhr, Wilma; Shen, Jianzhong; Schwarz, Stefan

    2017-02-01

    The multiresistance plasmid pAFS11, obtained from a bovine methicillin-resistant Staphylococcus aureus (MRSA) isolate, was completely sequenced and analysed for its structure and organisation. Moreover, the susceptibility to the heavy metals cadmium and copper was determined by broth macrodilution. The 49,189-bp plasmid harboured the apramycin resistance gene apmA, two copies of the macrolide/lincosamide/streptogramin B resistance gene erm(B) (both located on remnants of a truncated transposon Tn917), the kanamycin/neomycin resistance gene aadD, the tetracycline resistance gene tet(L) and the trimethoprim resistance gene dfrK. The latter three genes were part of a 7,284-bp segment which was bracketed by two copies of IS431. In addition, the cadmium resistance operon cadDX as well as the copper resistance genes copA and mco were located on the plasmid and mediated a reduced susceptibility to cadmium and copper. Moreover, a complete novel ica-like gene cluster of so far unknown genetic origin was detected on this plasmid. The ica-like gene cluster comprised four different genes whose products showed 64.4-76.9% homology to the Ica proteins known to be involved in biofilm formation of the S. aureus strains Mu50, Mu3 and N315. However, 96.2-99.4% homology was seen to proteins from S. sciuri NS1 indicating an S. sciuri origin. The finding of five different antibiotic resistance genes co-located on a plasmid with heavy metal resistance genes and an ica-like gene cluster is alarming. With the acquisition of this plasmid, antimicrobial multiresistance, heavy metal resistances and potential virulence properties may be co-selected and spread via a single horizontal gene transfer event. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Prevalence of aac(3-IIa gene among clinical isolates of uropathogenic Escherichia coli in Delfan, Lorestan

    Directory of Open Access Journals (Sweden)

    Somayeh Momeni Mofrad

    2013-09-01

    Full Text Available Backgrounds: Uropathogenic Escherichia coli strains are the predominant causative organisms of urinary tract infections (UTIs. Aminoglycosides are clinically useful antibiotics with bactericidal activity against this bacterium. The most common mechanism for resistance to these antibiotics are mediated through production of aminoglycoside modifying enzymes (AMEs. The most common of these enzymes are Aminoglycoside Acetyltransferases (AACs. The epidemiology of the dominant type of these enzymes, AAC(3-II, varies from region to region. The aim of this study was to determine the antimicrobial susceptibility pattern with a focus on aminoglycosides and the prevalence of aac(3-IIa gene among clinical isolates of uropathogenic Escherichia coli obtained from Delfan, Lorestan, Iran. Materials and Methods: In this descriptive study, a total of 100 uropathogenic Escherichia coli isolates were collected from BoAli hospital in Delfan city, Lorestan, from July to November 2010. Antibiotic susceptibility patterns of the isolates were determined using disk diffusion method according to Clinical and Laboratory Standards Institute CLSI guidelines. Prevalence of aac(3-IIa gene was determined by PCR and the relationship between resistance phenotypes to aminoglycosides and presence of aac(3-IIa gene was evaluated. Results: Among the 100 tested isolates, maximal resistance was seen to ampicillin (85%; whereas, no resistance to imipenem was found. Sixty percent of the isolates demonstrated resistance to at least one of the tested aminoglycosides. Resistance rate towards these agents were as followed: gentamicin 39%, kanamycin 26%, neomycin 31% and amikacin 1%. Forty–four isolates (44% harbored the aac(3-IIa gene. The maximal rate of gene presence (36 isolates, 92.3% was detected in strains with gentamicin resistant phenotype (39 isolates, 39%. Conclusion: On the basis of our findings, use of antibiotics such as nitrofurantoin, amikacin or imipenem are recommended for

  18. Longitudinal characterization of monophasic Salmonella Typhimurium throughout the pig's life cycle.

    Science.gov (United States)

    Fernandes, Laura; Centeno, Maria Madalena; Couto, Natacha; Nunes, Telmo; Almeida, Virgílio; Alban, Lis; Pomba, Constança

    2016-08-30

    Swine have been described as an important reservoir of multidrug resistant monophasic Salmonella Typhimurium, though information on its ecology is scarce. A longitudinal study was performed in order to elucidate the Salmonella 4,[5],12:i:- dynamics throughout the pig's production cycle. A total of 209 faecal samples were collected from 10 sows and in six sampling times during the life of 70 pigs from a Portuguese industrial farm, and 43 isolates of S. 4,[5],12:i:- were identified and characterized regarding clonality and antimicrobial resistance phenotype and genotype. Most isolates (n=42) exhibited resistance to at least ampicillin, kanamycin, neomycin, streptomycin, tetracycline and sulfonamides (encoded by blaTEM, aphAI-IAB, strA, strB, tetB and sul2, respectively). Isolates obtained during the finishing phase showed additional resistance to chloramphenicol and florfenicol (floR), gentamicin and netilmicin (aac(3')-IV). To our knowledge, this study is the first description of aphAI-IAB in S. 4,[5],12:i:-. PFGE analysis showed uneven distribution of isolates into three clusters, A (n=34), B (n=8) and C (n=1). PFGE cluster A was predominant in sows (n=5) and piglets in the farrowing phase (n=17) and in pigs in the early finishing phase (n=11) suggesting a carryover from birth to adult age. The introduction of PFGE cluster B isolates in adulthood could have had an external source, reinforcing the relevance of environmental transmission in the farm ecosystem. This study reveals a dynamic interaction between monophasic S. Typhimurium and the pressures exerted under an intensive swine production setting. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Functional activity of plasmid DNA after entry into the atmosphere of earth investigated by a new biomarker stability assay for ballistic spaceflight experiments.

    Directory of Open Access Journals (Sweden)

    Cora S Thiel

    Full Text Available Sounding rockets represent an excellent platform for testing the influence of space conditions during the passage of Earth's atmosphere and re-entry on biological, physical and chemical experiments for astrobiological purposes. We designed a robust functionality biomarker assay to analyze the biological effects of suborbital spaceflights prevailing during ballistic rocket flights. During the TEXUS-49 rocket mission in March 2011, artificial plasmid DNA carrying a fluorescent marker (enhanced green fluorescent protein: EGFP and an antibiotic resistance cassette (kanamycin/neomycin was attached on different positions of rocket exterior; (i circular every 90 degree on the outer surface concentrical of the payload, (ii in the grooves of screw heads located in between the surface application sites, and (iii on the surface of the bottom side of the payload. Temperature measurements showed two major peaks at 118 and 130 °C during the 780 seconds lasting flight on the inside of the recovery module, while outer gas temperatures of more than 1000 °C were estimated on the sample application locations. Directly after retrieval and return transport of the payload, the plasmid DNA samples were recovered. Subsequent analyses showed that DNA could be recovered from all application sites with a maximum of 53% in the grooves of the screw heads. We could further show that up to 35% of DNA retained its full biological function, i.e., mediating antibiotic resistance in bacteria and fluorescent marker expression in eukaryotic cells. These experiments show that our plasmid DNA biomarker assay is suitable to characterize the environmental conditions affecting DNA during an atmospheric transit and the re-entry and constitute the first report of the stability of DNA during hypervelocity atmospheric transit indicating that sounding rocket flights can be used to model the high-speed atmospheric entry of organics-laden artificial meteorites.

  20. Establishment of regeneration and transformation system in Egyptian sesame (Sesamum indicum L.) cv Sohag 1.

    Science.gov (United States)

    Al-Shafeay, Amal F; Ibrahim, Ahmed S; Nesiem, Mohamed R; Tawfik, Mohamed S

    2011-01-01

    Sesame (Sesamum indicum L.) is an important oil crop in many tropical and sub-tropical regions of the world, yet has received little attention in applying modern biotechnology in its improvement due to regeneration and transformation difficulties. Here within, we report the successful production of transgenic fertile plants of sesame (cv Sohag 1), after screening several cultivars. Agrobacterium tumefaciens- carrying the pBI121 plasmid {neomycin phosphotransferase gene (NPTII) and a β-glucuronidase gene (GUS)} was used in all experiments. Recovery of transgenic sesame shoots was achieved using shoot induction medium (Murashige and Skoog MS basal salt mixture + Gamborg's B5 vitamins + 2.0 mg/l BA + 1.0 mg/l IAA + 5.0 mg/l AgNO3 + 30.0 g/l sucrose + 7.0 g/l agar + 200 mg/l cefotaxime and 25 mg/l kanamycin) and shoots were rooted on MS medium + B5 vitamins + 1.0 mg/l IAA + 10.0 g/l sucrose and 7.0 g/l agar. Rooted shoots were transplanted into soil and grown to maturity in greenhouse. Incorporation and expression of the GUS gene into T0 sesame plants was confirmed using polymerase chain reaction (PCR), reverse transcriptase-PCR (RT-PCR) and GUS histochemical assay. Several factors were found to be important for regeneration and transformation in sesame. The most effective were plant genotype and the addition of AgNO3 for successful recovery of sesame shoots. Co-cultivation time and optical density of the Agrobacterium were also critical for sesame transformation. This work is an attempt to open the door for further genetic improvement of sesame using important agronomic traits.

  1. Characterization of Antibiotic Resistance Genes from Lactobacillus Isolated from Traditional Dairy Products.

    Science.gov (United States)

    Guo, Huiling; Pan, Lin; Li, Lina; Lu, Jie; Kwok, Laiyu; Menghe, Bilige; Zhang, Heping; Zhang, Wenyi

    2017-03-01

    Lactobacilli are widely used as starter cultures or probiotics in yoghurt, cheese, beer, wine, pickles, preserved food, and silage. They are generally recognized as safe (GRAS). However, recent studies have shown that some lactic acid bacteria (LAB) strains carry antibiotic resistance genes and are resistant to antibiotics. Some of them may even transfer their intrinsic antibiotic resistance genes to other LAB or pathogens via horizontal gene transfer, thus threatening human health. A total of 33 Lactobacillus strains was isolated from fermented milk collected from different areas of China. We analyzed (1) their levels of antibiotic resistance using a standardized dilution method, (2) their antibiotic resistance gene profiles by polymerase chain reaction (PCR) using gene-specific primers, and (3) the transferability of some of the detected resistance markers by a filter mating assay. All Lactobacillus strains were found to be resistant to vancomycin, but susceptible to gentamicin, linezolid, neomycin, erythromycin, and clindamycin. Their susceptibilities to tetracycline, kanamycin, ciprofloxacin, streptomycin, quinupristin/dalfopristin, trimethoprim, ampicillin, rifampicin, and chloramphenicol was different. Results from our PCR analysis revealed 19 vancomycin, 10 ciprofloxacin, and 1 tetracycline-resistant bacteria that carried the van(X), van(E), gyr(A), and tet(M) genes, respectively. Finally, no transferal of the monitored antibiotic resistance genes was observed in the filter mating assay. Taken together, our study generated the antibiotic resistance profiles of some milk-originated lactobacilli isolates and preliminarily assessed their risk of transferring antibiotic gene to other bacteria. The study may provide important data concerning the safe use of LAB. © 2017 Institute of Food Technologists®.

  2. [Detection of auditory impairment in the offsprings caused by drug treatment of the dams].

    Science.gov (United States)

    Kameyama, T; Nabeshima, T; Itoh, J

    1982-12-01

    To study the auditory impairment induced by prenatal administration of aminoglycosides in the offspring, the shuttle box method to measure the auditory threshold of rats (Kameyama et al., Folia pharmacol. japon. 77, 15, 1981) was employed. Four groups of pregnant rats were administered 200 mg/kg kanamycin sulfate (KM), 200 mg/kg dihydrostreptomycin sulfate (DHSM), 100 mg/kg neomycin sulfate (NM), or 1 ml/kg saline intramuscularly from the 10th to the 19th day of pregnancy. The auditory threshold of the offspring could be measured by the shuttle box method in about 90% of the live born rats at the age of 100 days. The auditory thresholds of the groups were as follows (mean +/- S.E.): saline group, 53.8 +/- 0.6 dB (N = 36); KM group, 63.8 +/- 1.1 dB (N = 34); DHSM group, 60.0 +/- 1.2 dB (N = 29); NM group, 62.4 +/- 1.2 dB (N = 24). Auditory thresholds of drug-treated groups were significantly higher than that of the saline group. However, no increase in the auditory threshold of the mother rat was detected after treatment with aminoglycosides. In addition, the experimental procedure of the shuttle box method is very easy, and the auditory threshold of a large number of rats could be measured in a short period. These findings suggest that this method is a very useful one for screening for auditory impairment induced by prenatal drug treatment in rat offspring.

  3. Antibiotic resistance in Salmonella Enteritidis isolated from broiler carcasses Resistência antimicrobiana em Salmonella Enteritidis isoladas de carcaças de frango

    Directory of Open Access Journals (Sweden)

    Martha Oliveira Cardoso

    2006-09-01

    Full Text Available Eighty Salmonella Enteritidis strains isolated from broiler carcasses between May 1995 and April 1996 in the State of Rio Grande do Sul, Brazil, were tested for antibiotic susceptibility using the disk diffusion method. Resistance to colistin, novobiocin, erythromycin and tetracycline was observed in 100% of the isolates. The strains showed intermediate resistance at different levels to kanamycin (1.25%, enrofloxacin (3.75%, neomycin (3.75%, fosfomycin (20%, sulphonamides (86.25% and nitrofurantoin (90%. Resistance to ciprofloxacin, norfloxacin, gentamicin, polymyxin B, sulphametrim and sulphazotrim was not found. Since resistance to antibiotics especially those introduced in the last decades, was detected, it is recommended that their use must be based on the results of resistance tests or minimum inhibitory concentration tests.Oitenta amostras de Salmonella Enteritidis isoladas de carcaças de frango no período entre maio de 1995 a abril de 1996 no Estado do Rio Grande do Sul, Brasil foram testados para susceptibilidade antimicrobiana pelo método de antibiograma. O antibiograma das amostras apresentou 100% de resistência a colistina, novobiocina, eritromicina e tetraciclina. Tiveram resistência em diferentes níveis a canamicina (1,25%, enrofloxacina (3,75%, neomicina (3,75%, fosfomicina (20%, sulfonamida (86,25% e nitrofurantoína (90% e por outro lado não apresentaram resistência a ciprofloxacina, norfloxacina, gentamicina, polimixina B, sulfametrim e sulfazotrim. A constatação de resistência a antibióticos, inclusive àqueles introduzidos na última década, enfatiza a necessidade de uso responsável de antibióticos, e com base em antibiograma ou concentração inibitória mínima.

  4. Scientific Opinion on a request from the European Commission for the assessment of the scientific elements put forward by Luxembourg to support the prohibition for the placing on the market of GM potato EH92-527-1 for cultivation purposes in Luxembourg

    Directory of Open Access Journals (Sweden)

    EFSA Panel on Genetically Modified Organisms (GMO

    2012-09-01

    Full Text Available

    Luxembourg notified to the European Commission its scientific arguments justifying the implementation of a national safeguard measure prohibiting the placing on the market of GM potato EH92-527-1 for cultivation purposes in Luxembourg, after which the European Commission asked the European Food Safety Authority (EFSA to assess the scientific information supporting the prohibition. Having considered the information package provided by Luxembourg and all relevant scientific publications, the EFSA Panel on Genetically Modified Organisms (GMO Panel concluded that: (i no new data specific to the safety of the nptII gene have been provided; (ii although bacterial DNA release and development of competence are expected to occur more efficiently in biofilms, the link between resistance in biofilms and cultivation/processing of GM potato EH92-527-1 was not established by Luxembourg, and the main barriers, limiting the transformation frequency of bacterial cells with transgenic plant DNA, remain; (iii the risk posed by the formation of mosaic structures of aminoglycoside phosphotransferase genes could not be assessed without data documenting the existence of such structures among the existing gene variants, and such data were not provided; (iv the knowledge gaps and uncertainties highlighted in the Luxembourgeois document and the therapeutic relevance of kanamycin and neomycin have already been considered in the previous EFSA opinion on antibiotic resistance marker genes, and no new information on the safety of nptII gene as present in the GM potato EH92-527-1 has been identified in the scientific literature that would cause the GMO Panel to change its previous conclusions. Therefore, the EFSA GMO Panel concludes that no grounds exist to date that would lead to reconsideration of its opinion on GM potato EH92-527-1.

  5. 链霉素抗性突变理性筛选新霉素高产菌株%Streptomycin-resistant Rational Screening for High-yield Neomycin Producing Strain

    Institute of Scientific and Technical Information of China (English)

    陈宏; 张祝兰; 孙菲; 张引; 周璟明; 郑榕

    2014-01-01

    采用化学诱变剂NTG结合链霉素抗性筛选法获得新霉素高产菌株.出发菌株费氏链霉菌(Streptomyces fradiae)FS1109的孢子悬液经不同剂量的化学诱变剂NTG处理后,涂布在含链霉素最小抑制浓度(3μg/mL)的培养基平板上培养,获得大量的链霉素抗性突变株.经影印法初筛和摇瓶发酵复筛,正突变率高于负突变率,获得一株遗传性状稳定的Streptomyces fradiae Str 63菌株,其新霉素生物活性单位比出发菌株提高了50%以上,且C组分较出发菌株的低.

  6. Study on Control Efficiency of Hydrochloride Neomycin 1% on Soil Borne Disease of Eggplant%1%申嗪霉素防治茄子土传病害药效试验

    Institute of Scientific and Technical Information of China (English)

    梁莉红

    2012-01-01

    于2010年秋季至2011年春季,进行1%申嗪霉素对茄子枯黄萎病的防治效果试验.结果表明:应用1%申嗪霉素防治茄子土传病害,以4.5g/m2覆膜效果为最佳,黄萎病发病率比对照低5.68%,增产5.77%,建议在生产上推广应用.

  7. 新得米先可溶性粉中新霉素测定方法的研究%Study on the determination of neomycin in Neo-Terramycin soluble powder

    Institute of Scientific and Technical Information of China (English)

    张秀英; 吴好庭

    2004-01-01

    利用盐酸土霉素能溶于无水乙醇而硫酸新霉素不溶的特性,将土霉素从新得米先可溶性粉中分离除去,再按抗生素微生物检定法测定新霉素的含量.该方法的平均回收率为100.3%,RSD为0.4%.按此方法测定3批供试品,含新霉素分别为标示量的92.7%、98.8%和98.9%.

  8. 薄层色谱法快速鉴别复方新霉素滴鼻液中三种主要成分%Qualitative Analysis of Compound Neomycin Nasal Drops By TLC

    Institute of Scientific and Technical Information of China (English)

    江霞; 陈怡

    2003-01-01

    采用薄层色谱法在同一薄层板上对复方新霉素滴鼻液中的三种主要成分硫酸新霉素、马来酸氯苯那敏与盐酸萘甲唑林进行定性鉴别,方法简便,易行,重现性好.

  9. Drug: D03262 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available pathologic organisms and parasites 61 Antibiotics 616 Acting mainly on acid-fast bacteria 6169 Others D03262...GENTS A07A INTESTINAL ANTIINFECTIVES A07AA Antibiotics A07AA08 Kanamycin D03262 Kanamycin sulfate (JP16) J A... ORGANS S01 OPHTHALMOLOGICALS S01A ANTIINFECTIVES S01AA Antibiotics S01AA24 Kanamycin D03262 Kanamycin sulfa

  10. Coenzyme Q10 protects hair cells against aminoglycoside.

    Directory of Open Access Journals (Sweden)

    Kazuma Sugahara

    Full Text Available It is well known that the production of free radicals is associated with sensory cell death induced by an aminoglycoside. Many researchers have reported that antioxidant reagents protect sensory cells in the inner ear, and coenzyme Q10 (CoQ10 is an antioxidant that is consumed as a health food in many countries. The purpose of this study was to investigate the role of CoQ10 in mammalian vestibular hair cell death induced by aminoglycoside. Cultured utricles of CBA/CaN mice were divided into three groups (control group, neomycin group, and neomycin + CoQ10 group. In the neomycin group, utricles were cultured with neomycin (1 mM to induce hair cell death. In the neomycin + CoQ10 group, utricles were cultured with neomycin and water-soluble CoQ10 (30-0.3 µM. Twenty-four hours after exposure to neomycin, the cultured tissues were fixed, and vestibular hair cells were labeled using an anti-calmodulin antibody. Significantly more hair cells survived in the neomycin + CoQ10 group than in the neomycin group. These data indicate that CoQ10 protects sensory hair cells against neomycin-induced death in the mammalian vestibular epithelium; therefore, CoQ10 may be useful as a protective drug in the inner ear.

  11. Targeting C-myc G-Quadruplex: Dual Recognition by Aminosugar-Bisbenzimidazoles with Varying Linker Lengths

    Directory of Open Access Journals (Sweden)

    Nihar Ranjan

    2013-11-01

    Full Text Available G-quadruplexes are therapeutically important biological targets. In this report, we present biophysical studies of neomycin-Hoechst 33258 conjugates binding to a G-quadruplex derived from the C-myc promoter sequence. Our studies indicate that conjugation of neomycin to a G-quadruplex binder, Hoechst 33258, enhances its binding. The enhancement in G-quadruplex binding of these conjugates varies with the length and composition of the linkers joining the neomycin and Hoechst 33258 units.

  12. Antibacterial activity of norfloxacin in the gastrointestinal tracts of rats.

    OpenAIRE

    Frimodt-Møller, P C; Jensen, K. M.; Madsen, P O

    1983-01-01

    The capacities of norfloxacin (MK-0366) and neomycin to reduce the numbers of bacteria in the gastrointestinal tracts of rats were evaluated. Results of a 3-day treatment with norfloxacin were compared with those of a 3-day treatment with neomycin. Both drugs significantly decreased gram-negative and gram-positive bacteria. Norfloxacin effected a significantly greater reduction in numbers of gram-negative bacteria than did neomycin. Norfloxacin also significantly increased the number of anaer...

  13. Ganciclovir

    Science.gov (United States)

    ... aminoglycoside antibiotics such as amikacin (Amikin), gentamicin (Garamycin), neomycin (New-Rx, New-Fradin), netilmicin (Netromycin), streptomycin, tobramycin (Nebcin, Tobi), and others; amphotericin B (Fungizone); ...

  14. Valganciclovir

    Science.gov (United States)

    ... aminoglycoside antibiotics such as amikacin (Amikin), gentamicin (Garamycin), neomycin (Neo-Rx, Neo-Fradin), netilmycin (Netromycin), streptomycin, tobramycin (Nebcin, Tobi), and others; amphotericin B (Fungizone); ...

  15. Drug: D01618 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available 27 Dental preparations 276 Antibiotics 2760 Antibiotics D01618 Fradiomycin sulfa...A07A INTESTINAL ANTIINFECTIVES A07AA Antibiotics A07AA01 Neomycin D01618 Fradiomycin sulfate (JP16); Neomyci...THROAT PREPARATIONS R02AB Antibiotics R02AB01 Neomycin D01618 Fradiomycin sulfate... (JP16); Neomycin sulfate (USP) S SENSORY ORGANS S01 OPHTHALMOLOGICALS S01A ANTIINFECTIVES S01AA Antibiotics...ibiotics for external use D01618 Fradiomycin sulfate (JP16); Neomycin sulfate (USP)

  16. Prevalenoe of Drug - Resistant Staphylococci in Teheran University Hospital Wards

    Directory of Open Access Journals (Sweden)

    F. Shafa

    1960-01-01

    Full Text Available 1 Fifty coagulase posittve strains of staphylococc~~ ~folated fr.o~ .the nose"nand wrist of Hospital nurses have been examined for sensltfvlty to pemcilhn, tetracyclines,"nchloramphenicol, dihydrostreptomycin, erythrorriycm, neomycin, kana.n:ycin,"nbacitracin, polymyxin-B and the triple sulfa. The percentages of fully sensittve strains at the present are as followe:                                       Erythromycin                                       100%"nNeomycin                                             78%"nKanamycin                                            78%"nChloramphenicol                                     68%"nDihydrostreptomycin                               52%"nPenicillin                                                18%"nTetracyclines                                         16%"nPolymyxin-B                                            1%"nTriple sulfa                                              0%"n2 The following topics have been discussed:"na The origin anr" mechanism of drug resistance"nb Cross-resistr.nee"nc The hospital epidemiology of Staphylococcus"nd The clinical implications of Staphylococcus drug-resistance

  17. Caracterização de bactérias dos gêneros Mima e Herella (Tribo Mimeae, DeBord, 1942: 1 Propriedade morfo-bioquímicas e sensibilidade aos antibióticos

    Directory of Open Access Journals (Sweden)

    Altair A. Zebral

    1971-01-01

    % lactose and irregularly utilize rhamnose and cellobiose; in the synthetic base media the strains produced acid from lactose and have the same sugar oxidations as described before. Mima polymorpha was oxidase-negative and failed to utilize the carbohidrates tested in either the complex nitrogenous media or the synthetic base media. Mima polymorpha var. oxidans was oxidase-positive and failed to utilize the carbohydrates tested. The Herellea vaginicola and Mima polymorpha was very susceptible to gabromycin, knamycin, neomycin, colistin and the former was very sensitive to chloranphenicol and rovamycin. Mima polymorpha var. oxidans presented high sensibility to kanamycin, neomycin, colistin, chloranphenicol and nalidixic acid. With the results obtained by the sensibility of strains to the 1 and 0,1 unity of penicillin by milliliter, it was impossible to separate the oxidase-positive and oxidase-negative strains in discordance with the data obtained by Baumann, Dodoroff & Stanier (1968a.

  18. First identification of methicillin-resistant Staphylococcus pseudintermedius strains among coagulase-positive staphylococci isolated from dogs with otitis externa in Trinidad, West Indies

    Directory of Open Access Journals (Sweden)

    Francis Dziva

    2015-12-01

    Full Text Available Background: Otitis externa is a common inflammatory ear disease in dogs caused by a variety of pathogens, and coagulase-positive staphylococci are frequently isolated from such infections. Objective: To identify antimicrobial susceptibility profiles and methicillin-resistant strains among coagulase-positive staphylococci isolated from otitis externa in dogs. Methods: A cross-sectional study was performed over 2 years on 114 client-owned dogs presented to the Veterinary Teaching Hospital with a primary complaint of ear infections. Swabs were obtained from both ears and cultured for staphylococci which were subsequently confirmed as coagulase-positive using rabbit plasma. Antimicrobial susceptibility assays were assessed on all isolates followed by subsequent genetic analysis for species identification and detection of the mecA gene. Results: Sixty-five coagulase-positive staphylococci were isolated from 114 client-owned dogs. The isolates exhibited resistance against neomycin (58.5%, streptomycin (49.2%, penicillin (49.2%, polymyxin B (44.6%, tetracycline (36.9%, sulphamethoxazole/trimethoprim (33.8%, kanamycin (33.8%, doxycycline (32.3%, norfloxacin (23.1%, amoxicillin/clavulanate (20%, ciprofloxacin (20%, enrofloxacin (18.5%, gentamicin (16.9%, and cephalothin (9.2%. Forty (61.5% of the isolates were resistant to at least three or more antimicrobials and 10 were sensitive to all. Using a multiplex polymerase chain reaction assay based on species-specific regions of the thermonuclease (nuc gene, 38/65 (58.5% isolates were classified as Staphylococcus aureus, 23/65 (35.4% as S. pseudintermedius, 2/65 (3.1% as S. intermedius, and 2/65 (3.1% as S. schleiferi. Analysis for the mecA gene revealed two positive isolates of S. pseudintermedius which were oxacillin-resistant, representing a first report of such organisms in the Caribbean. Conclusion: Despite the relatively high prevalence of multidrug-resistant coagulase-positive staphylococci in

  19. First identification of methicillin-resistant Staphylococcus pseudintermedius strains among coagulase-positive staphylococci isolated from dogs with otitis externa in Trinidad, West Indies

    Science.gov (United States)

    Dziva, Francis; Wint, Crystal; Auguste, Tennille; Heeraman, Carolyn; Dacon, Cherrelle; Yu, Priscilla; Koma, Lee M.

    2015-01-01

    Background Otitis externa is a common inflammatory ear disease in dogs caused by a variety of pathogens, and coagulase-positive staphylococci are frequently isolated from such infections. Objective To identify antimicrobial susceptibility profiles and methicillin-resistant strains among coagulase-positive staphylococci isolated from otitis externa in dogs. Methods A cross-sectional study was performed over 2 years on 114 client-owned dogs presented to the Veterinary Teaching Hospital with a primary complaint of ear infections. Swabs were obtained from both ears and cultured for staphylococci which were subsequently confirmed as coagulase-positive using rabbit plasma. Antimicrobial susceptibility assays were assessed on all isolates followed by subsequent genetic analysis for species identification and detection of the mecA gene. Results Sixty-five coagulase-positive staphylococci were isolated from 114 client-owned dogs. The isolates exhibited resistance against neomycin (58.5%), streptomycin (49.2%), penicillin (49.2%), polymyxin B (44.6%), tetracycline (36.9%), sulphamethoxazole/trimethoprim (33.8%), kanamycin (33.8%), doxycycline (32.3%), norfloxacin (23.1%), amoxicillin/clavulanate (20%), ciprofloxacin (20%), enrofloxacin (18.5%), gentamicin (16.9%), and cephalothin (9.2%). Forty (61.5%) of the isolates were resistant to at least three or more antimicrobials and 10 were sensitive to all. Using a multiplex polymerase chain reaction assay based on species-specific regions of the thermonuclease (nuc) gene, 38/65 (58.5%) isolates were classified as Staphylococcus aureus, 23/65 (35.4%) as S. pseudintermedius, 2/65 (3.1%) as S. intermedius, and 2/65 (3.1%) as S. schleiferi. Analysis for the mecA gene revealed two positive isolates of S. pseudintermedius which were oxacillin-resistant, representing a first report of such organisms in the Caribbean. Conclusion Despite the relatively high prevalence of multidrug-resistant coagulase-positive staphylococci in Trinidad

  20. Bacterial Analysis of Bovine Mastitis in Part Regions of Tianjin%天津部分地区奶牛乳房炎细菌学分析

    Institute of Scientific and Technical Information of China (English)

    李利山; 王晓敏; 刘燕霏; 杨建德

    2015-01-01

    乳房炎是奶牛业最常见且损失最为严重的疾病之一.为了解天津地区奶牛场乳房炎病原,本研究对天津地区4个奶牛场的77份乳样进行了病原菌的分离与鉴定分析.通过病原分离、细菌形态学观察、生化试验鉴定发现,临床型及亚临床型乳房炎乳样分离菌均以金黄色葡萄球菌和大肠杆菌为优势菌群.药敏试验结果表明,分离菌对环丙沙星、头孢他啶、氧氟沙星、复方新诺明、庆大霉素、卡那霉素、四环素和新霉素敏感,对其他抗菌素药物均呈现不同程度的耐药.本研究明确了天津地区奶牛乳房炎的主要病原,并筛选出了敏感药物,这对该病的防治具有指导意义.%Bovine mastitis is one of the most prevalent diseases in the cow industry with severe economical loss. To understand the etiology of bovine mastitis and provide references for controlling bovine mastitis in Tianjin, in this study, the suspected milk samples of bovine mastitis from four cow farms in this region were analyzed by pathogen isolation, morphology observation, biochemical tests and drug sensitivity tests. The results show that Staphylococcus aureus andE coli. Were the main isolates and they were sensitive to ciprofloxacin, ceftazidime, levofloxacin, cotrimoxazole, gentamicin, kanamycin, tetracycline and neomycin, with various degrees of resistance to other antibiotics. The study demonstrated the main pathogens and sensitive drugs for bovine mastitis in part regions of Tianjin, and provided guiding significance for prevention and control of the disease.

  1. 鹅胚沙门氏菌的药敏试验及消毒试验%Antibiotics Sensitivity and Disinfect Experiment of Goose Embryo Salmonella

    Institute of Scientific and Technical Information of China (English)

    湛琳丽; 周廷富

    2012-01-01

    采用纸片扩散法,对重庆市荣昌县某种鹅场死胚的9株鹅胚沙门氏茵,进行了13种抗菌药物的敏感性测定。还比较了7种消毒剂的抑茵强度,结果表明:9株茵对恩诺沙星、卡那霉素、氟哌酸高敏,对强力霉素、新霉素、庆大霉素、壮观霉素、氨苄青霉素中敏,而对复方磺胺、青霉素G、链霉素、洁霉素、四环素具有一定的耐药性;必灭杀可作为家禽防病灭痛的首选消毒药物,茵毒敌、复合酚、瑞得士203、甲酚皂、新洁而灭、畜禽乐消毒剂可作为家禽防病灭病的一般消毒药物。%With the Filter-paper diffusion method,for Rongchang county Chongqing city certain goose farm become sick the 9 goose embryo Salmonella that split in death chick, the sensitivity that has carried out 14 kinds of antibiotics, have still compared 7 kinds of disinfectant restrain bacterium strength, show as a result: 16 bacteriums are high sensitive to enrofloxaein, kanamycin, Norfloxacin, and are moderate sensitive to Doxycycline, Neomycin, Furazolidone, Gentamycin, Spectinomycin, Ampicillin, and have drug resistance to Compound sulfamethoxazole, Penicillin G, Streptomycin, Lincomycin, Tetracycline. Bromosept can be the first selection that extinguishes disease sterilized medicine.Farmot complex phenol, Rui De Shi Disinfectant Bactericide Contianing Double Chain Quatemaies and Saponated Cresol Solution , benzalkonium bromide, Chu Qin Ie (a New veterinary Medicine)can be used as general sterilized medicine for domestic animals.

  2. Isolation and Growth Characteristics of a Psychro-Tolerant Methanobacterium SHB%耐低温甲烷杆菌SHB的分离鉴定与生长特性分析

    Institute of Scientific and Technical Information of China (English)

    马金亮; 习彦花; 张丽萍; 王宏伟; 程辉彩

    2012-01-01

    利用亨盖特厌氧操作技术,从承德塞罕坝湿地污泥中分离出1株耐低温的产甲烷菌SHB.该菌株宽约0.31 μm,长约2.87 μm,细杆,形成长丝.能够利用H2/CO2和甲钠盐生长,不利用甲醇、三甲胺、乙酸和二级醇类.最适生长温度范围为16 ~40℃,最适pH为6.0 ~9.0,能在0% ~5%盐浓度范围内生长.该菌株对青霉素、链霉素、罗红霉素、利福平、新霉素有抗性,对氯霉素、四环素、卡那霉素、庆大霉素敏感.经生理、生化及16S rDNA分析,确定该菌株与甲烷杆菌属(Methanobacterium sp.)的亲缘关系最近.该菌株的生长温度和pH范围都很宽,在沼气生产方面有潜在的应用前景.%A psychro-tolerant methane-producing strain SHB was isolated from sludge collected in Saihanba wetland, Chengde, by hungate roll-tube technique. The strain was 0.31 μm μ2.87 μm, thin rod, forming filament. It could utilize formate and H2/CO2 as substrate but not acetate, methanol, trimethylamine, and/or secondary alcohols. The most suitable growth temperture ranged at 16 ~ 40 ℃ with pH 6 ~9, and could grow within 0% -5% of salt. The strain resisted to penicillin, streptomycin, roxithromycin, rifampicin, and neomycin, but sensitive to tetracycline, chloramphenicol, kanamycin, and genlamycin. Through physio-biochemical characteristics and 16S rDNA analyses the osculation of the strain was confirmed close the most to the genus of Methanobaclerium. The wide growth temperature and pH range of the strain made it potential application outlook in mathane production.

  3. Monitoring of multiple resistance of Streptococcus suis and Haemophilus parasuis strains from clinical isolates%临床分离猪链球菌和副猪嗜血杆菌多重耐药性监测

    Institute of Scientific and Technical Information of China (English)

    胡军勇; 汤细彪; 汤电; 刘望宏; 倪德斌; 吴斌

    2012-01-01

    为了解我国规模化猪场链球菌和副猪嗜血杆菌的耐药情况,用纸片扩散法对临床分离的112株猪链球菌和92株副猪嗜血杆菌进行23种药物的敏感性试验.结果表明,猪链球菌对林可胺类、大环内酯类、硝基呋喃类药物的耐药率均在90%以上,对链霉素、庆大霉素、卡那霉素、阿米卡星、多西环素的耐药率在60%~90%之间,而对β-内酰胺类和氟喹诺酮类的耐药率相对较低.副猪嗜血杆菌对四环素耐药率最高(78.3%),对螺旋霉素、复方磺胺甲(嗯)唑、克林霉素、阿米卡星、林可霉素、新霉素的耐药率在50%~70%之间,对头孢类和氟喹诺酮类耐药率相对较低.2种细菌对不同种类药物的敏感性各异,并且多重耐药性严重,主要耐药谱具有多样性.%One hundred and twelve Streptococcus suis and 92 Haemophilus parasuis were isolated from pig farm to investigate susceptibility of the two bacteria strains to 23 antimicrobials by using disk diffusion method. Results showed that for the S. Suis strain,the resistance rates of lincosamtde,macroli-des and nitrofurans were all above 90%,followed by streptomycin,gentamicin,kanamycin,amikacin and doxycycline with the resistance rates ranged from 60% to 90% and resistance rates of β-lactams and quinolones were relatively lower. For the H. Parasuis strain, the resistance rate of tetracycline was the highest (78. 3%), followed by spiramycin, sulfamethoxazole/TMP, clindymycin, lincomycin, amikacin and neomycin varied from 50% to 70% and resistance rates of cephalosporins and quinolones were relatively lower. The susceptibility of the two kinds of bacteria to the antimicrobials were different, multiple resistance was severe and the resistance profiles were various.

  4. Determination of aminoglycoside residues in milk and muscle based on a simple and fast extraction procedure followed by liquid chromatography coupled to tandem mass spectrometry and time of flight mass spectrometry.

    Science.gov (United States)

    Arsand, Juliana Bazzan; Jank, Louíse; Martins, Magda Targa; Hoff, Rodrigo Barcellos; Barreto, Fabiano; Pizzolato, Tânia Mara; Sirtori, Carla

    2016-07-01

    Antibiotics are widely used in veterinary medicine mainly for treatment and prevention of diseases. The aminoglycosides are one of the antibiotics classes that have been extensively employed in animal husbandry for the treatment of bacterial infections, but also as growth promotion. The European Union has issued strict Maximum Residue Levels (MRLs) for aminoglycosides in several animal origin products including bovine milk, bovine, swine and poultry muscle. This paper describes a fast and simple analytical method for the determination of ten aminoglycosides (spectinomycin, tobramycin, gentamicin, kanamycin, hygromycin, apramycin, streptomycin, dihydrostreptomycin, amikacin and neomycin) in bovine milk and bovine, swine and poultry muscle. For sample preparation, an extraction method was developed using trichloroacetic acid and clean up with low temperature precipitation and C18 bulk. Liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) was used to carry out quantitative analysis and liquid chromatography-quadrupole-time of flight-mass spectrometry (LC-QTOF-MS) was used to screening purposes. Both methods were validated according to the European Union Commission Directive 2002/657/EC. Good performance characteristics were obtained for recovery, precision, calibration curve, specificity, decision limits (CCα) and detection capabilities (CCβ) in all matrices evaluated. The detection limit (LOD) and quantification limit (LOQ) were ranging from 5 to 100ngg(-1) and 12.5 to 250ngg(-1), respectively. Good linearity (r)-above 0.99-was achieved in concentrations ranging from 0.0 to 2.0×MRL. Recoveries ranged from 36.8% to 98.0% and the coefficient of variation from 0.9 to 20.2%, noting that all curves have been made into their own matrices in order to minimize the matrix effects. The CCβ values obtained in qualitative method were between 25 and 250ngg(-1). The proposed method showed to be simple, easy, and adequate for high-throughput analysis of a large

  5. Two P5CS genes from common bean exhibiting different tolerance to salt stress in transgenic Arabidopsis

    Indian Academy of Sciences (India)

    Ji Bao Chen; Jian Wei Yang; Zhao Yuan Zhang; Xiao Fan Feng; Shu Min Wang

    2013-12-01

    Many plants accumulate proline in response to salt stress. -pyrroline-5-carboxylate synthetase (P5CS) is the rate-limiting enzyme in proline biosynthesis in plants. Plasmid DNA (pCHF3-PvP5CS1 and pCHF3-PvP5CS2) containing the selectable neomycin phosphotransferase gene for kanamycin resistance and Phaseolus vulgaris P5CS (PvP5CS1 and PvP5CS2) cDNA was introduced into Arabidopsis plants using Agrobacterium-mediated gene transfer. Southern blot, northern blot and RT-PCR analyses demonstrated that the foreign genes were integrated into Arabidopsis chromosomal DNA and expressed. Single-gene transformants were analysed in this study. Transgenic plants expressed higher levels of PvP5CS1 and PvP5CS2 transcripts under salt stress conditions than under normal conditions. When treated with 0, 100 and 200 mM NaCl, the average proline content in leaves of transgenic plants was significantly higher $(P \\lt 0.01)$ than control plants. The average relative electrical conductivity (REC) of transgenic lines was significantly lower $(P \\lt 0.01)$ than control plants under salt stress condition. Biomass production of transgenic lines was significantly higher $(P \\lt 0.05)$ than control plants under 200 mM NaCl stress treatment. These results indicated that introducing PvP5CS1 and PvP5CS2 cDNA into transgenic Arabidopsis caused proline overproduction, increasing salt tolerance. Although the expression of PvP5CS1 in L4 lines and PvP5CS2 in S4 lines was the same under salt stress condition, the S4 lines accumulated 1.6 and 1.9 times more proline than the L4 lines under 100 and 200 mM NaCl treatments, respectively. The REC of S4 plants was 0.5 (100 mM NaCl) and 0.6 times (200 mM NaCl) that of L4 plants. The biomass production of S4 plants was 1.6 times (200 mM NaCl) more than in L4 plants. Total P5CS enzyme activity of S4 was significantly higher than that of L4. These results implied that the PvP5CS2 protein had stronger capacity to catalyze proline synthesis than PvP5CS1 under salt

  6. Epidemiological Investigation of Swine Pathogenic E.coli in Guangxi Small and Medium-scale Farms%广西中小规模猪场大肠杆菌病流行情况调查

    Institute of Scientific and Technical Information of China (English)

    陈凤莲; 赵武; 黄红梅; 覃绍敏; 华俊; 马玲; 吴健敏; 杨威

    2011-01-01

    从广西23家中小规模猪场采集病料,分离、鉴定出致病性大肠杆菌56株,并对其进行血清型鉴定和耐药性检测,结果鉴定出43株致病性大肠杆菌,分属于16种血清型,其中以O138、O18、O85、O9和O21为优势血清型,占定型菌株的69.76%;用20种常用抗菌药进行药物敏感性试验,发现高敏的药物有头孢拉啶、头孢唑啉、头孢曲松、头孢西丁、多黏菌素B、新霉素、壮观霉素和阿米卡星;而不敏感的药物有青霉素、阿莫西林、强力霉素、卡那霉素、万古霉素、林可霉素.说明这些猪场大肠杆菌的耐药性十分严重,应加强生物安全措施等进行综合防控.%56 strains of swine pathogenic E.coli were isolated from 23 small and medium-scale pig farms in Guangxi, and its serotype identification and detection of drug resistance showed that the 56 strains belonged to 16 serotypes, out of which O serotype viz, O138 ,O18 ,O85 ,O9 ,O21 were predominant, occupying 69.76%.The detection of drug resistance of 20 frequently used antibiotics indicated that most E.coli trains were highly sensitive to Cephradine, Cefazolin, Ceftriaxone, Cefoxitin, Polymyxin B, Neomycin, Spectinomycin, and Amikacin, but insensitive to Penicillin, Amoxicillin, Doxycycline, Kanamycin, Vancomycin, Lincomycin, which points to the fact that drug resistance of E.coli in these pig farms has been very grave, and that measures such as biotic safety should be strengthened to undertake comprehensive prevention.

  7. Development of efficient catharanthus roseus regeneration and transformation system using agrobacterium tumefaciens and hypocotyls as explants

    Directory of Open Access Journals (Sweden)

    Wang Quan

    2012-06-01

    Full Text Available Abstract Background As a valuable medicinal plant, Madagascar periwinkle (Catharanthus roseus produces many terpenoid indole alkaloids (TIAs, such as vindoline, ajamlicine, serpentine, catharanthine, vinblastine and vincristine et al. Some of them are important components of drugs treating cancer and hypertension. However, the yields of these TIAs are low in wild-type plants, and the total chemical synthesis is impractical in large scale due to high-cost and their complicated structures. The recent development of metabolic engineering strategy offers a promising solution. In order to improve the production of TIAs in C. roseus, the establishment of an efficient genetic transformation method is required. Results To develop a genetic transformation method for C. roseus, Agrobacterium tumefaciens strain EHA105 was employed which harbors a binary vector pCAMBIA2301 containing a report β-glucuronidase (GUS gene and a selectable marker neomycin phosphotransferase II gene (NTPII. The influential factors were investigated systematically and the optimal transformation condition was achieved using hypocotyls as explants, including the sonication treatment of 10 min with 80 W, A. tumefaciens infection of 30 min and co-cultivation of 2 d in 1/2 MS medium containing 100 μM acetosyringone. With a series of selection in callus, shoot and root inducing kanamycin-containing resistance media, we successfully obtained stable transgenic regeneration plants. The expression of GUS gene was confirmed by histochemistry, polymerase chain reaction, and genomic southern blot analysis. To prove the efficiency of the established genetic transformation system, the rate-limiting gene in TIAs biosynthetic pathway, DAT, which encodes deacetylvindoline-4-O-acetyltransferase, was transferred into C. roseus using this established system and 9 independent transgenic plants were obtained. The results of metabolite analysis using high performance liquid chromatography (HPLC

  8. 磷酸二(2-乙基己基)酯-磷酸三丁酯-异辛烷反胶束萃取卡那霉素的研究%Study on Extraction of Kanamycin with HDEHP-TBP-Isooctane Reverse Micelles

    Institute of Scientific and Technical Information of China (English)

    李夏兰; 翁连进; 陈培钦; 蔡婀娜

    2006-01-01

    目的 研究反胶束提取氨基糖苷类抗生素的可能性,寻求高效且环境友好的氨基糖苷类抗生素的生产工艺.方法 将磷酸二(2-乙基己基)酯(HDEHP)、磷酸三丁酯(TBP)、异辛烷形成反胶束,与卡那霉素溶液或发酵滤液混合、振荡、离心.结果 反胶束萃取率与水相pH值、盐浓度密切相关.当KCl浓度为0.05 mol·L-1,萃取前水相pH为10,HDEHP浓度为1 mol·L-1时,HDEHP-TBP-异辛烷反胶束萃取发酵滤液的萃取率为88.6%.结论 HDEHP-TBP-异辛烷反胶束萃取反胶束是可行的.

  9. Production of Axenic Gonyaulax Cultures by Treatment with Antibiotics

    OpenAIRE

    1982-01-01

    The effects of amphotericin B, chloramphenicol, dihydrostreptomycin sulfate, neomycin sulfate, polymyxin B sulfate, potassium penicillin G, and streptomycin sulfate (used singularly and in various combinations at different concentrations) on the growth and development of four marine dinoflagellates of the genus Gonyaulax and associated bacteria were studied. The combination of amphotericin B, dihydrostreptomycin, neomycin, and penicillin G was highly effective in eliminating bacteria and fung...

  10. 77 FR 64715 - New Animal Drugs; Approvals; Changes of Sponsor; Change of Sponsor's Name; Change of Sponsor's...

    Science.gov (United States)

    2012-10-23

    ... rights and interest in, NADA 065-015 for VETROPOLYCIN HC (bacitracin zinc, polymyxin B sulfate, neomycin sulfate, and hydrocortisone) Ophthalmic Ointment, NADA 065-016 for VETROPOLYCIN (bacitracin zinc, neomycin... narasin Type C medicated swine feeds. (d) * * * (1) * * * (xi) * * * (B) Limitations. For broiler chickens...

  11. A library screening approach identifies naturally occurring RNA sequences for a G-quadruplex binding ligand.

    Science.gov (United States)

    Mirihana Arachchilage, Gayan; Morris, Mark J; Basu, Soumitra

    2014-02-07

    An RNA G-quadruplex library was synthesised and screened against kanamycin A as the ligand. Naturally occurring G-quadruplex forming sequences that differentially bind to kanamycin A were identified and characterized. This provides a simple and effective strategy for identification of potential intracellular G-quadruplex targets for a ligand.

  12. Determination of clobetasol propionate and miconazole nitrate in clobetasol propionate, miconazole nitrate,neomycin sulfate and urea cream by HPLC%HPLC法测定索咪新尿素乳膏中丙酸氯倍他索和硝酸咪康唑的含量

    Institute of Scientific and Technical Information of China (English)

    王希东

    2011-01-01

    目的 采用高效波相色谱法(HPLC)测定索咪新尿素乳膏中丙酸氯倍他索和硝酸咪康唑的含量.方法 采用Diamonsil C18柱(150 mm×4.6 mm,5μm)为色谱柱,以甲醇-0.1%醋酸铵溶液(80:20)为流动相,以苯甲酸苄酯为内标,流速为1.0 mL/min,紫外检测器于240 nm测定.结果 HPLC法测定,样品与基质分离良好,丙酸氯倍他索和硝酸咪康唑的线性范围分别为7.5~12.5 mg/L和0.3 ~ 0.5 mg/mL内(r=0.9999),平均回收率均>98.00%,样品溶液在48 h内稳定.结论 HPLC法简便、准确、灵敏、重现性好,能有效控制索咪新尿素乳膏的内在质量.

  13. Determination of clobetasol propionate and miconazole nitrate in clobetasol propionate,miconazole nitrate,neomycin sulfate and urea cream by HPLC%HPLC法测定索咪新尿素乳膏中丙酸氯倍他索和硝酸咪康唑的含量

    Institute of Scientific and Technical Information of China (English)

    郭燕萍; 郁韫超; 施孝金; 钟明康

    2010-01-01

    目的 建立HPLC法同时测定索咪新尿素乳膏中丙酸氯倍他索和硝酸咪康唑的含量.方法 采用Diamonsil C18柱,流动相为甲醇-0.1%乙酸铵溶液(90:10,V/V),流速1.0mL·nin-1,检测波长240nm,内标为苯甲酸苄酯.结果 丙酸氯倍他索在7.5~12.5 mg·L-1内和硝酸咪康唑在300~500 mg·L-1内线性关系良好(r=0.999 9),回收率均>98%,日内、日间RSD均<1.5%.结论 本方法准确灵敏,可用于索咪新尿素乳膏中丙酸氯倍他索和硝酸咪康唑的含量测定.

  14. Estudo comparativo de duas técnicas farmacopéicas de avaliação da atividade antimicrobiana dos fármacos: nistatina, eritromicina, neomicina e gentamicina Comparison of two pharmacopeical thecnics to the evaluation of the antimicrobial activity of the drugs: nystatin, erythromycin, neomycin and gentamicin

    OpenAIRE

    Tatiane Margato Vital; Cleomenes Reis; Marco Túlio Antonio García-Zapata; Luiz Carlos da Cunha

    2004-01-01

    Através das técnicas de difusão em ágar com discos de papel e cilindros de aço inoxidável, analisou-se quantitativamente a atividade antimicrobiana de 123 medicamentos contendo os fármacos: nistatina (43 amostras de creme vaginal), eritromicina (14 amostras de comprimidos e 9 amostras de suspensão oral), gentamicina (33 amostras de líquido injetável) e neomicina (24 amostras de creme de uso tópico), mediante as seguintes cepas de microrganismos: Saccharomyces cerevisiae ATCC 2601 para a nista...

  15. Role of activity of gastrointestinal microflora in absorption of calcium and magnesium in rats fed beta1-4 linked galactooligosaccharides.

    Science.gov (United States)

    Chonan, O; Takahashi, R; Watanuki, M

    2001-08-01

    Rats fed a diet containing beta1-4 linked galactooligosaccharides (GOS) (5 g/100 g of diet) absorbed calcium and magnesium more efficiently than those fed the control diet. However, the increment obtained through GOS-feeding was reduced by neomycin sulfate (0.67 g/100 g of diet). Since the decrease in cecal pH in rats fed GOS was suppressed by neomycin-feeding, bacterial action in the digestive tract was considered to be reduced by neomycin-feeding. Our findings suggest that the action of intestinal bacteria is necessary for the effects of GOS.

  16. Drug: D07994 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available AGENTS A07A INTESTINAL ANTIINFECTIVES A07AA Antibiotics A07AA01 Neomycin D07994 Framycetin sulfate B BLOOD A...CALS S01A ANTIINFECTIVES S01AA Antibiotics S01AA03 Neomycin D07994 Framycetin sulfate S01AA07 Framycetin D07...R01AX08 Framycetin D07994 Framycetin sulfate R02 THROAT PREPARATIONS R02A THROAT PREPARATIONS R02AB Antibi...otics R02AB01 Neomycin D07994 Framycetin sulfate S SENSORY ORGANS S01 OPHTHALMOLOGI

  17. Varicose and other vein problems - self-care

    Science.gov (United States)

    ... neomycin Drying lotions, such as calamine Lanolin, a natural moisturizer Benzocaine or other creams that numb the skin Watch for skin sores on your leg, especially around your ankle. Take care of sores right away to prevent infection. When ...

  18. Characterisation of recently emerged multiple antibiotic-resistant Salmonella enterica serovar typhimurium DT104 and other multiresistant phage types from Danish pig herds

    DEFF Research Database (Denmark)

    Baggesen, Dorte Lau; Aarestrup, Frank Møller

    1998-01-01

    A total of 670 isolates of Salmonella enterica were isolated from Danish pig herds, phage typed and tested for susceptibility to amoxycillin + clavulanate, ampicillin, colistin, enrofloxacin, gentamicin, neomycin, spectinomycin, streptomycin, tetracyclines, and trimethoprim + sulphadiazine. S ent...

  19. Associations between baseline allergens and polysensitization

    DEFF Research Database (Denmark)

    Carlsen, Berit Christina; Menné, Torkil; Johansen, Jeanne Duus

    2008-01-01

    : Seven allergens--parabens mix, N-isopropyl-N-phenyl-p-phenylenediamine, sesquiterpene lactone mix, wool alcohols, potassium dichromate, Myroxylon pereirae, and cobalt chloride - showed statistically significant positive associations to polysensitization. Five allergens p-phenylenediamine, neomycin...

  20. Associations between baseline allergens and polysensitization

    DEFF Research Database (Denmark)

    Carlsen, B.C.; Menne, T.; Johansen, J.D.

    2008-01-01

    : Seven allergens - parabens mix, N-isopropyl-N-phenyl-p-phenylenediamine, sesquiterpene lactone mix, wool alcohols, potassium dichromate, Myroxylon pereirae, and cobalt chloride - showed statistically significant positive associations to polysensitization. Five allergens p-phenylenediamine, neomycin...

  1. Combined therapy of Salmonella infection in chickens by antimicrobial agents followed by cultured cecal bacteria.

    Science.gov (United States)

    Seuna, E; Schneitz, C; Nurmi, E

    1980-06-01

    Week-old chickens infected with Salmonella infantis when one day old were treated with antimicrobial drugs either given alone or followed by peroral inoculation of bacterial culture. The bacteria were derived from the cecal contents of adult chickens. The antimicrobial drugs used were: neomycin, neomycin plus oxytetracycline, neomycin plus polymyxin, and sulfadiazine plus trimethoprim. The combined therapy with oxytetracycline plus neomycin and bacterial culture seemed to be the most effective, although the efficacy varied between the parallel trials. Sulfadiazine plus trimethoprim followed by treatment with the bacterial culture decreased the infection rate. The bacterial culture alone also had a slight anti-salmonella effect. When only antimicrobials were given, salmonellae rapidly reappeared in the intestines when the therapy was stopped.

  2. Oral antibiotics enhance antibody responses to keyhole limpet hemocyanin in orally but not muscularly immunized chickens

    National Research Council Canada - National Science Library

    Murai, Atsushi; Kitahara, Kazuki; Okumura, Shouta; Kobayashi, Misato; Horio, Fumihiko

    2016-01-01

    .... We aimed to determine whether the modification of gut microbiota by oral co‐administration of two antibiotics, ampicillin and neomycin, would lead to changes in the antibody response to antigens in chickens...

  3. Synergistic Antibacterial Effect of Silver Nanoparticles Combined with Ineffective Antibiotics on Drug Resistant Salmonella typhimurium DT104.

    Science.gov (United States)

    McShan, Danielle; Zhang, Ying; Deng, Hua; Ray, Paresh C; Yu, Hongtao

    2015-01-01

    Synergistic antibacterial activity of combined silver nanoparticles (AgNPs) with tetracycline (polykeptide), neomycin (aminoglycoside), and penicillin (β-lactam) was tested against the multidrug resistant bacterium Salmonella typhimurium DT104. Dose-dependent inhibition of Salmonella typhimurium DT104 growth is observed for tetracycline-AgNPs and neomycin-AgNPs combination with IC50 of 0.07 μg/mL and 0.43 μg/mL, respectively. There is no inhibition by the penicillin-AgNPs combination. These results suggest that the combination of the ineffective tetracycline or neomycin with AgNPs effectively inhibits the growth of this bacterium. The synergistic antibacterial effect is likely due to enhanced bacterial binding by AgNPs assisted by tetracycline or neomycin, but not by penicillin.

  4. Maintaining Toxic Tort Claims against the Federal Government: A Case Study of Liability Theories Defenses and Remedies

    Science.gov (United States)

    1990-02-18

    issue is United States v. Kubrick . 2 Plaintiff received neomycin treatment in 1968 and later developed a hearing loss. In January 1969, a physician...advised Ji.t was highly possible (or likely?) this druq was the cause. While pursuing a VA claim in 1971, Kubrick was told neomycin caused his injuries...These comments merely prompt suspicion. Although he knew the cause within two years of filing, the Court said Kubrick could have discovered it earlier. 33

  5. Antibiotic Manipulation of Intestinal Microbiota To Identify Microbes Associated with Campylobacter jejuni Exclusion in Poultry▿ †

    OpenAIRE

    Scupham, A. J.; Jones, J. A.; Rettedal, E. A.; Weber, T. E.

    2010-01-01

    The ability of various subsets of poultry intestinal microbiota to protect turkeys from colonization by Campylobacter jejuni was investigated. Community subsets were generated in vivo by inoculation of day-old poults with the cecal contents of a Campylobacter-free adult turkey, followed by treatment with one antimicrobial, either virginiamycin, enrofloxacin, neomycin, or vancomycin. The C. jejuni loads of the enrofloxacin-, neomycin-, and vancomycin-derived communities were decreased by 1 log...

  6. Chromatid Paints: A New Method for Detecting Tumor-Specific Chromosomal Inversions

    Science.gov (United States)

    1999-10-01

    a neomycin resistance marker (NEO) by homologous recombination in the yeast Saccha- have been established and previously described [14]. romyces...irslSF-C7 mutant knockout in DT40 chicken lymphoblastoid cells, line. This implies that, while XRCC2 and XRCC3 shutdown of RAD51 expression resulted in...Morita, H. Yamada, M. Oshimura, Construction of mouse A9 clones containing a single human chromosome tagged with neomycin -resistance gene via mi- [1

  7. Comparison of the sensitivities of the Buehler test and the guinea pig maximization test for predictive testing of contact allergy

    DEFF Research Database (Denmark)

    Frankild, S; Vølund, A; Wahlberg, J E;

    2001-01-01

    dose-response model. To compare the sensitivity of the 2 test procedures the test conditions were kept identical and the following chemicals with a range of sensitization potentials were tested: chloraniline, chlorhexidine, eugenol, formaldehyde, mercaptobenzothiazole and neomycin sulphate....... Formaldehyde and neomycin sulphate were strong sensitizers in both tests. Mercaptobenzothiazole, eugenol and chloraniline were all strong sensitizers in the GPMT, eugenol and mercaptobenzothiazole were negative in the Buehler test and equivocal results were obtained with chloraniline. Chlorhexidine...

  8. Comparison of the efficacy of diclofenac and betamethasone following strabismus surgery

    OpenAIRE

    Wright, M.; Butt, Z; McIlwaine, G; Fleck, B.

    1997-01-01

    AIMS—To compare the relative anti-inflammatory potency and safety of topical diclofenac-gentamicin with beta methasone-neomycin following strabismus surgery.
METHODS—A single centre, single observer, prospective, randomised, and double masked clinical trial of 25 children undergoing bilateral symmetrical horizontal strabismus surgery was carried out. One eye received diclofenac-gentamicin and the contralateral eye received betamethasone-neomycin; both treatments were instilled four times a da...

  9. Drug: D05140 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available RRHEALS, INTESTINAL ANTIINFLAMMATORY/ANTIINFECTIVE AGENTS A07A INTESTINAL ANTIINFECTIVES A07AA Antibiotics A...N) R02 THROAT PREPARATIONS R02A THROAT PREPARATIONS R02AB Antibiotics R02AB01 Neomycin D05140 Framycetin (IN...N) S SENSORY ORGANS S01 OPHTHALMOLOGICALS S01A ANTIINFECTIVES S01AA Antibiotics S01AA03 Neomycin D05140 Fram

  10. Determination of ten aminoglycoside residues in milk and dairy products using high performance liquid chromatography-tandem mass spectrometry%高效液相色谱-串联质谱法检测乳制品中10种氨基糖苷类抗生素残留

    Institute of Scientific and Technical Information of China (English)

    龚强; 丁利; 朱绍华; 焦艳娜; 成婧; 付善良; 王利兵

    2012-01-01

    建立了乳制品中链霉素、双氢链霉素、新霉素、卡那霉素、妥布霉素、庆大霉素、安普霉素、潮霉素B、巴龙霉素、阿米卡星等10种氨基糖苷类抗生素(aminoglycosides,AGs)残留的高效液相色谱-串联质谱(HPLC-MS/MS)检测方法.乳制品提取液经亲水-亲脂平衡(hydrophilic-lipophilic balance,HLB)柱净化后,采用反相离子对高效液相色谱分离,电喷雾串联四极杆质谱检测.对样品前处理条件、液相色谱流动相以及质谱条件进行了优化.结果表明:10种AGs在20 ~1000 μg/L范围内定量离子的峰面积和样品的质量浓度之间有很好的线性关系;在乳制品中的加标回收率为71.2%~101.7%,相对标准偏差为3.4%~13.8%.该方法简便、灵敏、准确,可用于乳制品中多种AGs残留的同时检测.%A high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/ MS) analytical method was developed for the simultaneous determination of ten aminoglycoside residues (streptomycin, dihydrostrepmycin, neomycin, kanamycin, tobramycin, gentamycin, apramycin, hygromycin B, paromomycin, and amkacin) in milk and dairy products. The sample was extracted with 5% trichloroacetic acid aqueous solution, then the extract was purified by a hydrophilic-lipophilic balance (HLB) cartridge. The ten aminoglycoside residues were separated by ion-pair reversed phase high performance liquid chromatography. Heptafluorobutyric acid was used as ion pair agent due to its volatility. Then the analytes were detected by electro-spray ionization tandem mass spectrometry. The pretreatment condition of the sample, the HPLC condition and the MS operation parameters were optimized. The results showed that the linearities of the ten aminoglycoside residues in 20 - 1 000 |xg/L had the correlation coefficient between 0. 994 6 -0.999 7. The recoveries ranged from 71.2% and 101.7% with the relative standard deviations of 3.4% - 13.8%. The proposed method was

  11. Antibiotic Susceptibility of Lactobacillus bulgaricus Isolated from Yoghurt%酸奶中保加利亚乳杆菌药物敏感性分析

    Institute of Scientific and Technical Information of China (English)

    周宁; 张建新; 樊明涛; 王静; 李子龙; 魏新元

    2012-01-01

    Lactobacillus bulgaricus strains were isolated from different yoghurt brands produced in different regions in China and analyzed for genetic diversity and antibiotic susceptibility.Further,antibiotic-resistant isolates were subjected to genetic analysis.A total of 18 Lactobacillus bulgaricus strains were obtained using modified MRS medium.They were subjected to RAPD typing and analyzed for susceptibility to 11 different antibiotics.Besides,their potential antibiotic-resistance genes were detected by PCR.The results showed that 18 Lactobacillus bulgaricus isolates had considerable genetic diversity and diverse antibiotic-resistance phenotypes.All of them were susceptible to roxithromycin and resistant to kanamycin.Meanwhile,they also could resist ampicillin,penicillin G,chlortetracycline,chloramphenicol,tetracycline,lincomycin,streptomycin,neomycin and gentamycin to different extents.tet(M) gene was detected in isolate B-8,and ant(6) gene in both B-8 and B-41,and aph(3')-Ⅲa gene in B-43,B-47,B-49 and B-51.These findings suggested that 18 Lactobacillus bulgaricus isolates had serious multiple antibiotic resistance.%从酸奶中分离保加利亚乳杆菌,对其遗传多样性和药物敏感性进行分析,并进一步对耐药菌株的抗性基因进行检测。利用改良MRS培养基,从国内不同品牌酸奶中分离获得18株保加利亚乳杆菌。18株分离菌先经RAPD分型后采用琼脂稀释法测定其对11种抗生素的药敏性,并通过PCR对耐药菌株中可能存在的抗性基因进行检测。结果显示,18株受试菌具有明显的遗传多样性和耐药表型多样性。18株菌全部对罗红霉素敏感,而全部对卡那霉素耐药;对氨苄青霉素、青霉素G、金霉素、氯霉素、四环素、林克霉素、链霉素、新霉素及庆大霉素等9种抗生素均表现出不同程度的耐药性。通过检测耐药菌株的抗性基因,从1株菌(B-8)中检出四环素抗性基因tet(M),从2株菌(B-8和B-41)

  12. 一株正二十二烷降解菌的分离及鉴定%Isolation and Identification on A Strain of N-docosane Degrading Bacteria

    Institute of Scientific and Technical Information of China (English)

    谭洪; 彭超; 唐赟; 刘亮; 杨艳

    2011-01-01

    [目的]研究正二十二烷降解菌的分离及鉴定.[方法]以正二十二烷无机盐培养基为选择培养基,从南充市炼油厂曝气池的回流污泥中筛选出1株高效降解长链烷烃的菌株,命名为T2,并对其进行形态学观察、生理生化鉴定和16S rDNA比对以及17种药物敏感试验.[结果]T2菌株鉴定为沙雷氏菌属.在正二十二烷浓度为1%(W/V)无机盐培养基中接入1%种子液,并在30℃和180 r/min摇瓶震荡下培养6d,正二十二烷降解率可达70%.药敏试验表明,T2菌株对链霉素、卡拉霉素、大观霉、氯霉素、氧氟沙星、庆大霉素、恩诺沙星、新霉素、阿米卡星、复方新诺明高度敏感;对环丙沙星为中度敏感;对青霉素G、氨苄青霉素、四环素、乙酰螺旋霉素、克林霉素、阿莫西林不敏感.[结论]该研究为石油污染的生物处理提供良好的微生物菌种资源.%[Objective] The aim was to study the isolation and identification of n-docosane degrading bacteria. [ Method ] With the mineral salt medium with the n-docosane as the selecting medium, a strain that could degrade the long-chain alkane effectively was screen out from the return sludge in the aeration pool of Nanchong Oil Refinery Plant, named as T2, and was conducted for the morphological observation, physiological and biochemical identification and 16S rDNA comparison, and the sensitive test with 17 kinds of drugs. [Result] The strain T2 was identified as Ser-ratia sp. The N-docosane degradation was 70% after 1% seed liquid of the strain was inoculated on the mineral salt medium with 1% ( W/V) of n-docosane and cultured at 30 ℃ and 180 r/min for 6 d. The drug the sensitivity tests showed that the T2 strain was sensitive to Streptomycin, Kanamycin, Spectinomycin, Chloromycetin, Ofloxacin, Gentamicin, Enrofloxacin, Neomycin, Amikaci and Sulfamethoxazole; intermediately sensitive to Ciprofloxacin; not sensitive to Penicillin G, Ampicillin, Tetracycline

  13. 黄喉拟水龟白眼病病原菌分离鉴定及药敏试验%Isolation, identification and drug sensitivity test of pathogen causing withe eye disease of Mauremys mutica

    Institute of Scientific and Technical Information of China (English)

    陆专灵; 钟一治; 赵忠添; 雷燕; 韦友传; 张益峰

    2016-01-01

    . mutica. The isolated strain was identified to be a gram-nega-tive bacillus with facultative anaerobic, acidogenicity, aerogenesis, fermentation and no exercise ability. The 16S rDNA sequences of the isolated strain ( GenBank accession number: KU855372 ) and that of Klebsiella pneumoniae had 99.1% homology. The isolated strain was hypersensitive to enrofloxacin, cephalothin, cefotaxime and ofloxacin, but showed drug resistance to streptomycin, neomycin, penicillin, kanamycin, tetracycline, clindamycin, doxycycline, amoxicillin and compound sulfamethoxazole. [Conclusion]K. pneumoniae is one of the main pathogenic bacteria causing withe eye disease of M. mutica. Quinolone and cephalosporin analogues such as enrofloxacin, ofloxacin cephalothin and cefotaxime can be used to prevent and control the disease.

  14. Evaluation of ghee based formulation for wound healing activity.

    Science.gov (United States)

    Prasad, Vure; Dorle, Avinash Kumar

    2006-08-11

    Formulation containing neomycin and ghee was evaluated for wound-healing potential on different experimental models of wounds in rats. The rats were divided into six groups of group 1 as control, group 2 as treated with neomycin only, group 3 as treated only with ghee, group 4 treated with F-1 formulation containing ghee 40% and neomycin 0.5%, group 5 treated with F-2 formulation containing ghee 50% and neomycin 0.5% and group 6 treated with F-3 formulation containing ghee and ointment base in all two wound models, each group consisting of six rats. Wound contraction ability in excision wound model was measured at different time intervals and study was continued until wound is completely healed. Tensile strength was measured in 10-day-old incision wound and quantitative estimation of hydroxy proline content in the healed tissue was determined in 10-day-old excision wound. Histological studies were done on 10-day-old sections of regenerated tissue of incision wound. F-2 formulation containing ghee 50% and neomycin 0.5% showed statistically significant response, in terms of wound contracting ability, wound closure time, period of epithelization, tensile strength of the wound, regeneration of tissues at wound site when compared with the control group and these results were comparable to those of a reference neomycin ointment.

  15. In vitro Synergy and Time-kill Assessment of Interaction between ...

    African Journals Online (AJOL)

    Pharmacotherapy Group, Faculty of Pharmacy, University of Benin, Benin City, 300001 .... µl aliquot was removed from the culture medium .... patients, multiple combinations of antibacterial .... therapeutic potential and safety of kanamycin-.

  16. Streptomycin interference in Jaffe reaction - Possible false positive creatinine estimation in excessive dose exposure

    DEFF Research Database (Denmark)

    Syal, Kirtimaan; Srinivasan, Anand; Banerjee, Dibyajyoti

    2013-01-01

    Objectives: To study the potential of commonly used aminoglycoside antibiotics to form non-creatinine chromogen with alkaline picrate reagent. Design and methods: We studied the non-creatinine chromogen formation of various concentrations of streptomycin, amikacin, kanamycin, netilmicin, gentamicin...

  17. Characterization of putative multidrug resistance transporters of the major facilitator-superfamily expressed in Salmonella Typhi

    DEFF Research Database (Denmark)

    Shaheen, Aqsa; Ismat, Fouzia; Iqbal, Mazhar

    2015-01-01

    conferred resistance to at least ten of the tested antimicrobials: ciprofloxacin, norfloxacin, levofloxacin, kanamycin, streptomycin, gentamycin, nalidixic acid, chloramphenicol, ethidium bromide, and acriflavine, including fluoroquinolone antibiotics, which were drugs of choice to treat S. Typhi infections...

  18. [Labelling of nif-plasmid pEA9 from Enterobacter agglomerans 339].

    Science.gov (United States)

    Liu, Cheng-jun; Klingmüller, Walter

    2002-07-01

    The authors describe the in vivo labelling of the plasmid pEA9 in Enterobacter agglomerans 339 with a kanamycin resistance gene. For labelling purposes the donor plasmid pST5 was constructed. This plasmid contains the nif ENX region from pEA9,in which a kanamycin resistance gene is cloned.pST5 was transformed into E.a.339 and subsequently cured from the host. Curing was achieved with AP medium. Fourty strains that had lost pST5,but retained the kanamycin resistance, could be isolated. It showed that none of these clones contained co-integrates of pST5 and pEA9. This is evident that in all clones the kanamycin resistance gene was integrated into pEA9 by homologous recombination.

  19. Innovations in Wound Infection Prevention and Management and Antimicrobial Countermeasures

    Science.gov (United States)

    2011-01-24

    therapies to treat wound infections – Acinetobacter baumannii , Pseudomonas aeruginosa, methicillin-resistant Staphylococcus aureus, extended...conventional antibiotics – converted Acinetobacter baumannii once resistant to kanamycin & gentamicin to susceptibility – Compound in Phase I clinical testing

  20. Tomato protoplast DNA transformation : physical linkage and recombination of exogenous DNA sequences

    NARCIS (Netherlands)

    Jongsma, Maarten; Koornneef, Maarten; Zabel, Pim; Hille, Jacques

    1987-01-01

    Tomato protoplasts have been transformed with plasmid DNA's, containing a chimeric kanamycin resistance gene and putative tomato origins of replication. A calcium phosphate-DNA mediated transformation procedure was employed in combination with either polyethylene glycol or polyvinyl alcohol. There