Exploring the Applicability of Cisplatin Uptake Inhibitors in the Reduction/Prevention of Ototoxicity

Introduction: Cisplatin is a chemotherapeutic agent used to treat many solid tumors, including ovarian, testicular, bladder, lung, and head and neck tumors. The main side effects associated with the drug include nephrotoxicity, peripheral neurotoxicity, and ototoxicity, with ototoxicity being the dose-limiting effect. Objective: The study\u27s overall goal is to determine if cimetidine, a cisplatin uptake inhibitor, has protective effects against cisplatin cytotoxicity on hearing cells with minimal reduction in cisplatin\u27s chemotherapeutic effects on cancer cells. To accomplish this, we want to first determine cisplatin\u27s dose- and time-dependent effects on HEI-OC1 cell viability. Method: House Ear Institute-Organ of Corti 1 (HEI-OC1) cells were grown in 6-well plates and treated 24 hours later with cisplatin concentrations of either 0, 10, 50, 100, 150, or 200uM. Cells were imaged using an inverted microscope at 48 hours. The number of dead and alive cells was counted using an automated cell counter at 72 hours. Results: Increasing levels of cisplatin concentrations resulted in decreasing numbers of HEI-OC1 cells that were alive and showed changes to cell morphology. Conclusion: Cisplatin exerts a negative effect on HEI-OC1 cell viability. We will utilize the above-identified condition to treat cancer and HEI-OC1 cells side by side with and without cimetidine

Variation in genome content and predatory phenotypes between \u3cem\u3eBdellovibrio\u3c/em\u3e sp. NC01 isolated from soil and \u3cem\u3eB. bacteriovorus\u3c/em\u3e type strain HD100

Defining phenotypic and associated genotypic variation among Bdellovibrio may further our understanding of how this genus attacks and kills different Gram-negative bacteria. We isolated Bdellovibrio sp. NC01 from soil. Analysis of 16S rRNA gene sequences and average amino acid identity showed that NC01 belongs to a different species than the type species bacteriovorus. By clustering amino acid sequences from completely sequenced Bdellovibrio and comparing the resulting orthologue groups to a previously published analysis, we defined a ‘core genome’ of 778 protein-coding genes and identified four protein-coding genes that appeared to be missing only in NC01. To determine how horizontal gene transfer (HGT) may have impacted NC01 genome evolution, we performed genome-wide comparisons of Bdellovibrio nucleotide sequences, which indicated that eight NC01 genomic regions were likely acquired by HGT. To investigate how genome variation may impact predation, we compared protein-coding gene content between NC01 and the B. bacteriovorus type strain HD100, focusing on genes implicated as important in successful killing of prey. Of these, NC01 is missing ten genes that may play roles in lytic activity during predation. Compared to HD100, NC01 kills fewer tested prey strains and kills Escherichia coli ML35 less efficiently. NC01 causes a smaller log reduction in ML35, after which the prey population recovers and the NC01 population decreases. In addition, NC01 forms turbid plaques on lawns of E. coli ML35, in contrast to clear plaques formed by HD100. Linking phenotypic variation in interactions between Bdellovibrio and Gram-negative bacteria with underlying Bdellovibrio genome variation is valuable for understanding the ecological significance of predatory bacteria and evaluating their effectiveness in clinical applications

Variation in genome content and predatory phenotypes between Bdellovibrio sp. NC01 isolated from soil and B. bacteriovorus type strain HD100

© The Author(s), 2019. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Williams, L. E., Cullen, N., DeGiorgis, J. A., Martinez, K. J., Mellone, J., Oser, M., Wang, J., & Zhang, Y. Variation in genome content and predatory phenotypes between Bdellovibrio sp. NC01 isolated from soil and B. bacteriovorus type strain HD100. Microbiology, 165(12), (2019): 1315-1330, doi: 10.1099/mic.0.000861.Defining phenotypic and associated genotypic variation among Bdellovibrio may further our understanding of how this genus attacks and kills different Gram-negative bacteria. We isolated Bdellovibrio sp. NC01 from soil. Analysis of 16S rRNA gene sequences and average amino acid identity showed that NC01 belongs to a different species than the type species bacteriovorus. By clustering amino acid sequences from completely sequenced Bdellovibrio and comparing the resulting orthologue groups to a previously published analysis, we defined a ‘core genome’ of 778 protein-coding genes and identified four protein-coding genes that appeared to be missing only in NC01. To determine how horizontal gene transfer (HGT) may have impacted NC01 genome evolution, we performed genome-wide comparisons of Bdellovibrio nucleotide sequences, which indicated that eight NC01 genomic regions were likely acquired by HGT. To investigate how genome variation may impact predation, we compared protein-coding gene content between NC01 and the B. bacteriovorus type strain HD100, focusing on genes implicated as important in successful killing of prey. Of these, NC01 is missing ten genes that may play roles in lytic activity during predation. Compared to HD100, NC01 kills fewer tested prey strains and kills Escherichia coli ML35 less efficiently. NC01 causes a smaller log reduction in ML35, after which the prey population recovers and the NC01 population decreases. In addition, NC01 forms turbid plaques on lawns of E. coli ML35, in contrast to clear plaques formed by HD100. Linking phenotypic variation in interactions between Bdellovibrio and Gram-negative bacteria with underlying Bdellovibrio genome variation is valuable for understanding the ecological significance of predatory bacteria and evaluating their effectiveness in clinical applications.This research was supported by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under grant no. P20GM103430 and by funding to LEW from Providence College. This material is based upon work conducted at a Rhode Island NSF EPSCoR research facility, the Genomics and Sequencing Center, supported in part by the National Science Foundation EPSCoR Cooperative Agreement #EPS-1004057. This material is based upon work supported in part by the National Science Foundation under EPSCoR Cooperative Agreement #OIA-1655221. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication