Project description:Aminoglycosides remain in current use because of the benefits, including the broad-spectrum, inexpensiveness to produce, and a relatively small incidence of allergic reactions. However, aminoglycosides may induce irreversible hearing loss. Recently, tacrine, an acetylcholineesterase inhibitor originally developed to treat Alzheimer’s disease, was found to be protective to neomycin-induced hair cell damage in zebrafish and mouse. However, the protection mechanism remain to be largely unknown. In this study, we revealed that tacrine significantly suppressed the expression of tnf mRNA and ROS production induced by neomycin. Reduction of tnf expression by the antisense morpholino or transcription activator-like effectors nuclease significantly suppressed the hair cell damage but not ROS production induced by neomycin. In zebrafish treated with an antagonist of NMDA receptor MK-801 or a mitochondrial ROS inhibitor acetyl-L-carnitine, ROS production, tnf expression, and hair cell damage induced by neomycin were also significantly reduced. These levels of suppression were not significantly different from those of co-treatment of tacrine and MK-801 or acetyl-L-carnitine. These findings suggest that activation of NMDA receptor followed by ROS production and subsequent tnf expression are involved in neomycin-induced hair cell damage and that tacrine ameliorate neomycin-induced hair cell damage through inhibition of the signaling pathway.
Project description:Much of human hearing loss is caused by loss of auditory hair cell (HC) function. Mammals cannot regenerate these essential mechanoelectrical transducers of sound. However, birds have retained the ability to regenerate HCs from surrounding supporting cells. To better understand hair cell regeneration, we have expression profiled the sensory epithelia from chicken cochleae and utricles. Pure sensory epithelia, consisting of HCs plus supporting cells, were damaged with either neomycin or laser treatment. Changes in gene expression at various points during regeneration were compared to undamaged control cultures on a custom microarray that interrogates the vast majority of transcription factor (TF) genes. These experiments involved multiple biological samples and hundreds of microarray comparisons. Keywords: timecourse
Project description:Much of human hearing loss is caused by loss of auditory hair cell (HC) function. Mammals cannot regenerate these essential mechanoelectrical transducers of sound. However, birds have retained the ability to regenerate HCs from surrounding supporting cells. To better understand hair cell regeneration, we have expression profiled the sensory epithelia from chicken cochleae and utricles. Pure sensory epithelia, consisting of HCs plus supporting cells, were damaged with either neomycin or laser treatment. Changes in gene expression at various points during regeneration were compared to undamaged control cultures on a custom microarray that interrogates the vast majority of transcription factor (TF) genes. These experiments involved multiple biological samples and hundreds of microarray comparisons. Keywords: timecourse
Project description:U2OS cells preferentially utilise integrin alphaV beta5 in adhesion complexes when grown under standard cell culture conditions for 3 days. AlphaV beta5 can be found in both classical 'Focal' adhesions and in novel 'Reticular' adhesions that do not associate with F-actin or other known adhesion protein components. In order to identify components of reticular adhesions, cells were treated with cytochalasinD in order to disrupt F-actin and promote loss of focal adhesions. Remaining reticular adhesion complexes were stabilised with DTBP crosslinking reagent before adhesion complexes were isolated. Preliminary data suggested that depletion of PIP2 by Neomycin treatment would lead to disruption of reticular adhesions preferentially over focal adhesions and so this manipulation was included in these experiments.
Project description:This randomized clinical trial studies how well neomycin and metronidazole hydrochloride with or without polyethylene glycol work in reducing infection in patients undergoing elective colorectal surgery. Polyethylene glycol, may draw water from the body into the colon, flushing out the contents of the colon. Antibiotics, like neomycin and metronidazole hydrochloride, may stop bacteria from growing. It is not yet known whether it’s better to give preoperative neomycin and metronidazole hydrochloride with or without polyethylene glycol in reducing surgical site infection after colorectal surgery.
Project description:(Part 1) Gene expression profiles of C. elegans in response to P. aeruginosa. Synchronized larval stage 1 (L1) worms were raised on E. coli OP50 for 72 hours. These synchronized young adult (YA) animals were subsequently exposed to P. aeruginosa PA14 for 4 hours. 25dC. (Part 2) Gene expression profiles of C. elegans in response to Neomycin/Streptomycin-mediated recovery after a 4 hour exposure to P. aeruginosa. Synchronized larval stage 1 (L1) worms were raised on E. coli OP50 for 72 hours. These synchronized young adult (YA) animals were subsequently exposed to P. aeruginosa PA14 for 4 hours and then treated with Neomycin and shifted to E. coli OP50 plus Streptomycin plates for 6, 12, or 24 hours to resolve the infection. As a control for Neomycin/Streptomycin exposure, synchronized larval stage 1 (L1) worms were raised on E. coli OP50 for 72 hours and either shifted to fresh E. coli OP50 plates for 6 hours or treated with Neomycin and shifted to E. coli OP50 plus Streptomycin plates for 6 hours. 25dC.
Project description:In order to obtain a global view of energy metabolism pathways of the sulfate-reducer Desulfovibrio vulgaris Hildenborough and the proteins involved therein whole-genome microarrays were used to compare the transcriptional response of cells grown with hydrogen/sulfate, pyruvate/sulfate, lactate/thiosulfate, and pyruvate with limiting sulfate, relative to growth in standard lactate/sulfate condition. Growth with hydrogen/sulfate showed the largest number of differently expressed genes and the largest changes in expression levels. The most up-regulated energy metabolism genes were those coding for the periplasmic [NiFeSe] hydrogenase, followed by the Ech hydrogenase, and the most down-regulated were genes coding for the Coo hydrogenase. The results point to the involvement of formate cycling and the ethanol pathway during growth on hydrogen, whereas there is evidence for CO cycling during growth on lactate and pyruvate, but not on H2. Growth with thiosulfate showed the hallmarks of a reduced energy status of the cells with down regulation of the ATP synthase and the Qmo and Dsr complexes., whereas growth with pyruvate showed the smallest differences but an increased role for the Ech hydrogenase.that in this case functions in reverse from the case of growth with H2. The multiple periplasmic hydrogenases and formate dehydrogenases, do not display the same regulation pattern showing that their metabolic roles are not totally interchangeable. This result together with the observation that several genes coding for proteins that have not been biochemically characterised were considerably affected in this study, reveals a more complex energy metabolism than previously considered and provides guidance for further studies. Keywords: Growth protocol
Project description:In order to obtain a global view of energy metabolism pathways of the sulfate-reducer Desulfovibrio vulgaris Hildenborough and the proteins involved therein whole-genome microarrays were used to compare the transcriptional response of cells grown with hydrogen/sulfate, pyruvate/sulfate, lactate/thiosulfate, and pyruvate with limiting sulfate, relative to growth in standard lactate/sulfate condition. Growth with hydrogen/sulfate showed the largest number of differently expressed genes and the largest changes in expression levels. The most up-regulated energy metabolism genes were those coding for the periplasmic [NiFeSe] hydrogenase, followed by the Ech hydrogenase, and the most down-regulated were genes coding for the Coo hydrogenase. The results point to the involvement of formate cycling and the ethanol pathway during growth on hydrogen, whereas there is evidence for CO cycling during growth on lactate and pyruvate, but not on H2. Growth with thiosulfate showed the hallmarks of a reduced energy status of the cells with down regulation of the ATP synthase and the Qmo and Dsr complexes., whereas growth with pyruvate showed the smallest differences but an increased role for the Ech hydrogenase.that in this case functions in reverse from the case of growth with H2. The multiple periplasmic hydrogenases and formate dehydrogenases, do not display the same regulation pattern showing that their metabolic roles are not totally interchangeable. This result together with the observation that several genes coding for proteins that have not been biochemically characterised were considerably affected in this study, reveals a more complex energy metabolism than previously considered and provides guidance for further studies. Keywords: Growth protocol Desulfovibrio vulgaris Hildenborough was cultured at 37°C with thiosulfate as the only electron acceptor (with lactate as the electron donor) to mid-log phase. Cultures were also cultivated similarly using sulfate as the sole electron acceptor to mid-log phase. Gene expression profiles of cultures grown with thiosulfate as the electron acceptor were compared with those of the cultures grown with sulfate as the electron acceptor for dissimilatory sulfate reductiontion. Total RNA was harvested from four replicate cultures for microarray analysis. RNA extraction, purification, and labeling were performed independently on each cell sample. Two samples of each total RNA preparation were labeled, one with Cy3-dUTP and another with Cy5-dUTP for microarray hybridization (dye swap).