Biology Commons^™

A Rationally Designed Synthetic Antimicrobial Peptide Against Pseudomonas-Associated Corneal Keratitis: Structure-Function Correlation, Sk Abdul Mohid, Prerana Sharma, Amani Alghalayini, Tripti Saini, Debarun Datta, Mark D. P. Willcox, Haydar Ali, Sreyan Raha, Achintya Singha, Nirakar Sahoo

Biology Faculty Publications and Presentations

Contact lens wearers are at an increased risk of developing Pseudomonas-associated corneal keratitis, which can lead to a host of serious ocular complications. Despite the use of topical antibiotics, ocular infections remain a major clinical problem, and a strategy to avoid Pseudomonas-associated microbial keratitis is urgently required. The hybrid peptide VR18 (VARGWGRKCPLFGKNKSR) was designed to have enhanced antimicrobial properties in the fight against Pseudomonas-induced microbial keratitis, including contact lens-related keratitis. In this paper, VR18's modes of action against Pseudomonas membranes were shown by live cell Raman spectroscopy, live cell NMR, live-cell fluorescence microscopy and measures taken using sparsely tethered bilayer …

Evolved Resistance To A Novel Cationic Peptide Antibiotic Requires High Mutation Supply, Alfonso Santos-Lopez, Melissa J. Fritz, Jeffrey B. Lombardo, Ansen H.P. Burr, Victoria A. Heinrich, Christopher Marshall, Vaughn S. Cooper

Biological Sciences Faculty Research and Publications

Background and Objectives

A key strategy for resolving the antibiotic resistance crisis is the development of new drugs with antimicrobial properties. The engineered cationic antimicrobial peptide WLBU2 (also known as PLG0206) is a promising broad-spectrum antimicrobial compound that has completed Phase I clinical studies. It has activity against Gram-negative and Gram-positive bacteria including infections associated with biofilm. No definitive mechanisms of resistance to WLBU2 have been identified.

Methodology

Here, we used experimental evolution under different levels of mutation supply and whole genome sequencing (WGS) to detect the genetic pathways and probable mechanisms of resistance to this peptide. We propagated populations …

Carbapenemase-Producing Pseudomonas Aeruginosa – An Emerging Challenge, Fred C. Tenover, David P. Nicolau, Christian M. Gill

Biology Faculty Publications

Carbapenem-resistant Pseudomonas aeruginosa (CR-PA) is a major healthcare-associated pathogen worldwide. In the United States, 10–30% of P. aeruginosa isolates are carbapenem-resistant, while globally the percentage varies considerably. A subset of carbapenem-resistant P. aeruginosa isolates harbour carbapenemases, although due in part to limited screening for these enzymes in clinical laboratories, the actual percentage is unknown. Carbapenemase-mediated carbapenem resistance in P. aeruginosa is a significant concern as it greatly limits the choice of anti-infective strategies, although detecting carbapenemase-producing P. aeruginosa in the clinical laboratory can be challenging. Such organisms also have been associated with nosocomial spread requiring infection prevention interventions. The carbapenemases …

Investigating The Antibacterial Activity Of Novel Metal Complexes Against Antimicrobial Resistant Pathogens, Megan O'Shaughnessy

Theses, Doctoral

Antimicrobial resistance (AMR) is one of the serious global health challenges of our time. There is an urgent need to develop novel therapeutic agents to overcome AMR, preferably through alternative mechanistic pathways from conventional treatments. Interdisciplinary research in inorganic medicinal chemistry with biology is advancing the knowledge and implementation of transition metal complexes for therapy and is offering a realistic alternative to traditional antibiotics. Metal complexes with 1,10-phenanthroline (phen) ligands have demonstrated promising therapeutic capabilities with diverse biological activity. Consequently, there has been a resurgence in research of these complexes as possible alternatives or adjuvants to established antimicrobial clinical therapeutics. …

In Vivo Activity Of Metal Complexes Containing 1,10-Phenanthroline And 3,6,9-Trioxaundecanedioate Ligands Against Pseudomonas Aeruginosa Infection In Galleria Mellonella Larvae, Megan O'Shaughnessy, Magdalena Piatek, Pauraic Mccarron, Malachy Mccann, Michael Devereux, Kevin Kavanagh, Orla L. Howe

Articles

Drug-resistant Pseudomonas aeruginosa is rapidly developing resulting in a serious global threat. Immunocompromised patients are specifically at risk, especially those with cystic fibrosis (CF). Novel metal complexes incorporating 1,10-phenanthroline (phen) ligands have previously demonstrated antibacterial and anti-biofilm effects against resistant P. aeruginosa from CF patients in vitro. Herein, we present the in vivo efficacy of {[Cu(3,6,9-tdda)(phen)₂]·₃H₂O·EtOH}_n (Cu-tdda-phen), {[Mn(3,6,9-tdda)(phen)₂]·₃H₂O·EtOH}_n (Mn-tdda-phen) and [Ag₂(3,6,9-tdda)(phen)₄]·EtOH (Ag-tdda-phen) (tddaH₂ = 3,6,9-trioxaundecanedioic acid). Individual treatments of these metal-tdda-phen complexes and in combination with the established antibiotic gentamicin were …

Periodically Disturbing The Spatial Structure Of Biofilms Can Affect The Production Of An Essential Virulence Factor In Pseudomonas Aeruginosa, Rebecca J. Quinn, Ivana Barraza, Laura García-Diéguez, Camryn Pajon, Lauren E. Krausfeldt, Kerollos Ibrahim, Laura A. Enzinna, Morgan E. Thorn, Omar Tonsi Eldakar, Travis J. A. Craddock, Robert P. Smith

Biology Faculty Articles

Understanding the environmental factors that affect the production of virulence factors has major implications in evolution and medicine. While spatial structure is important in virulence factor production, observations of this relationship have occurred in undisturbed or continuously disturbed environments. However, natural environments are subject to periodic fluctuations, including changes in physical forces, which could alter the spatial structure of bacterial populations and impact virulence factor production. Using Pseudomonas aeruginosa PA14, we periodically applied a physical force to biofilms and examined production of pyoverdine. Intermediate frequencies of disturbance reduced the amount of pyoverdine produced compared to undisturbed or frequently disturbed conditions. …

Phenotypic/Genotypic Profile Of Oxa-10-Like-Harboring, Carbapenem-Resistant Pseudomonas Aeruginosa: Using Validated Pharmacokinetic/Pharmacodynamic In Vivo Models To Further Evaluate Enzyme Functionality And Clinical Implications, Fred C. Tenover, Christian M. Gill, Adrian Brink, Chun Yat Chu, Jennifer Coetzee, George Dimopoulos, Clinton Moodley, Christoffel Johannes Opperman, Spyros Pournaras, Isabella A. Tickler, Hafsah Deepa Tootla, Sophia Vourli, David P. Nicolau

Biology Faculty Publications

In vitro MICs and in vivo pharmacodynamics of ceftazidime and cefepime human-simulated regimens (HSR) against modified carbapenem inactivation method (mCIM)-positive Pseudomonas aeruginosa isolates harboring different OXA-10-like subtypes were described. The murine thigh model assessed ceftazidime (2 g every 8 h [q8h] HSR) and cefepime (2 g and 1 g q8h HSR). Phenotypes were similar despite possessing OXA-10-like subtypes with differing spectra. Ceftazidime produced ≥1-log10 killing in all isolates. Cefepime activity was dose dependent and MIC driven. This approach may be useful in assessing the implications of β-lactamase variants.

Evaluation Of The Xpert Carba-R Nxg Assay For Detection Of Carbapenemase Genes In A Global Challenge Set Of Pseudomonas Aeruginosa Isolates, Fred C. Tenover, Christian M. Gill, Tomefa E. Asempa, Isabella A. Tickler, Caitlin M. Dela Cruz, David P. Nicolau

Biology Faculty Publications

The growing prevalence and diversity of carbapenemase producers among carbapenem-resistant Pseudomonas aeruginosa (CRPA) isolates warrants an expansion of detection capabilities. The purpose of this study was to evaluate the performance of the commercially available Xpert Carba-R (Carba-R) and the research-use-only Xpert Carba-R NxG (Carba-R NxG) in a global collection of P. aeruginosa. The challenge set included 123 P. aeruginosa clinical isolates from 12 countries. Isolates were previously categorized via PCR or whole-genome sequencing. Carbapenemase classes tested include VIM, IMP, NDM, SPM, KPC, and GES. Non-carbapenemase (non-CP)-harboring isolates were also tested (negative control). Isolates were tested using the Carba-R NxG and …

Draft Genome Sequences Of Two Pseudomonas Aeruginosa Isolates From The Female Urogenital Tract, Genevieve Johnson, Carine R. Mores, Alan J. Wolfe, Catherine Putonti

Bioinformatics Faculty Publications

Pseudomonas aeruginosa is a Gram-negative bacterium that has the ability to survive in and readily adapt to a variety of environmental conditions. Here, we report 2 genome sequences of P. aeruginosa strains, UMB1046 and UMB5686, isolated from the female urogenital tract.

Characterization Of The Φctx-Like Pseudomonas Aeruginosa Phage Dobby Isolated From The Kidney Stone Microbiota, Genevieve Johnson, Alan J. Wolfe, Catherine Putonti

Biology: Faculty Publications and Other Works

Bacteriophages (phages) are vital members of the human microbiota. They are abundant even within low biomass niches of the human body, including the lower urinary tract. While several prior studies have cultured bacteria from kidney stones, this is the first study to explore phages within the kidney stone microbiota. Here we report Dobby, a temperate phage isolated from a strain of Pseudomonas aeruginosa cultured from a kidney stone. Dobby is capable of lysing clinical P. aeruginosa strains within our collection from the urinary tract. Sequencing was performed producing a 37 152 bp genome that closely resembles the temperate P. aeruginosa …

Characterization Of The Φctx-Like Pseudomonas Aeruginosa Phage Dobby Isolated From The Kidney Stone Microbiota, Genevieve Johnson, Alan J. Wolfe, Catherine Putonti

Bioinformatics Faculty Publications

Bacteriophages (phages) are vital members of the human microbiota. They are abundant even within low biomass niches of the human body, including the lower urinary tract. While several prior studies have cultured bacteria from kidney stones, this is the first study to explore phages within the kidney stone microbiota. Here we report Dobby, a temperate phage isolated from a strain of Pseudomonas aeruginosa cultured from a kidney stone. Dobby is capable of lysing clinical P. aeruginosa strains within our collection from the urinary tract. Sequencing was performed producing a 37 152 bp genome that closely resembles the temperate P. aeruginosa …

Disarming Pseudomonas Aeruginosa Virulence By The Inhibitory Action Of 1,10-Phenanthroline-5,6-Dione- Based Compounds: Elastase B (Lasb) As A Chemotherapeutic Target, Anna Clara M. Galdino, Livia Vigano, Aleaxandre A. De Castron, Elaine F.F; Da Cunha, Thais P. Mello, Larissa M. Mattos, Marcos D. Pereira, Mary Hunt, Megan O'Shaughnessy, Orla L. Howe, Michael Devereux, Malachy Mccann, Teodorico C. Ramalho, Marta H. Branquinha, André L.S. Santos

Articles

negative pathogen Pseudomonas aeruginosa, and this enzyme orchestrates several physiopathological events during bacteria-host interplays. LasB is considered to be a potential target for the development of an innovative chemotherapeutic approach, especially against multidrug-resistant strains. Recently, our group showed that 1,10-phenanthroline-5,6-dione (phendione), [Ag(phendione)2]ClO4 (Ag-phendione) and [Cu(phendione)3](ClO4)2.4H2O (Cu-phendione) had anti-P. aeruginosa action against both planktonic- and biofilm-growing cells. In the present work, we have evaluated the effects of these compounds on the (i) interaction with the lasB active site using in silico approaches, (ii) lasB proteolytic activity by using a specific fluorogenic peptide substrate, (iii) lasB gene expression by real time-polymerase chain …

Deconvoluting The Effects Of Surface Chemistry And Nanoscale Topography: Pseudomonas Aeruginosa Biofilm Nucleation On Si-Based Substrates, Zhang Zhang, Jingling Huang, Carmen Say, Robert L. Dorit, Kate Queeney

Chemistry: Faculty Publications

Hypothesis

The nucleation of biofilms is known to be affected by both the chemistry and topography of the underlying substrate, particularly when topography includes nanoscale (nm) features. However, determining the role of topography vs. chemistry is complicated by concomitant variation in both as a result of typical surface modification techniques. Analyzing the behavior of biofilm-forming bacteria exposed to surfaces with systematic, independent variation of both topography and surface chemistry should allow differentiation of the two effects.

Experiments

Silicon surfaces with reproducible nanotopography were created by anisotropic etching in deoxygenated water. Surface chemistry was varied independently to create hydrophilic (OH- terminated) …

The 'Pseudomonas Aeruginosa' Psl Polysaccharide Is A Social But Noncheatable Trait In Biofilms, Yasuhiko Irie, Aled E. Roberts, Kasper N. Kragh, Vernita D. Gordon, Jaime B. Hutchison, Rosalind J. Allen, Gavin Melaugh, Thomas Bjarnsholt, Stuart A. West, Stephen P. Diggle

Biology Faculty Publications

Extracellular polysaccharides are compounds secreted by microorganisms into the surrounding environment, and they are important for surface attachment and maintaining structural integrity within biofilms. The social nature of many extracellular polysaccharides remains unclear, and it has been suggested that they could function as either cooperative public goods or as traits that provide a competitive advantage. Here, we empirically tested the cooperative nature of the PSL polysaccharide, which is crucial for the formation of biofilms in Pseudomonas aeruginosa. We show that (i) PSL is not metabolically costly to produce; (ii) PSL provides populationlevel benefits in biofilms, for both growth and antibiotic …

Environmental Pseudomonads Inhibit Cystic Fibrosis Patient-Derived Pseudomonas Aeruginosa, Payel Chatterjee, Elizabeth Davis, Fengan Yu, Sarah James, Julia H. Wildschutte, Daniel D. Wiegmann, David H. Sherman, Robert M. Mckay, John J. Lipuma, Hans Wildschutte

Great Lakes Institute for Environmental Research Publications

Pseudomonas aeruginosa is an opportunistic pathogen which is evolving resistance to many currently used antibiotics. While much research has been devoted to the roles of pathogenic P. aeruginosa in cystic fibrosis (CF) patients, less is known of its ecological properties. P. aeruginosa dominates the lungs during chronic infection in CF patients, yet its abundance in some environments is less than that of other diverse groups of pseudomonads. Here, we sought to determine if clinical isolates of P. aeruginosa are vulnerable to environmental pseudomonads that dominate soil and water habitats in one-to-one competitions which may provide a source of inhibitory factors. …

Elucidating The Role Of Mifs-Mifr Two-Component System In Regulating Pseudomonas Aeruginosa Pathogenicity, Gorakh Digambar Tatke

FIU Electronic Theses and Dissertations

Pseudomonas aeruginosa is a Gram-negative, metabolically versatile, opportunistic pathogen that exhibits a multitude of virulence factors, and is extraordinarily resistant to a gamut of clinically significant antibiotics. This ability is in part mediated by two-component systems (TCS) that play a crucial role in regulating virulence mechanisms, metabolism and antibiotic resistance. Our sequence analysis of the P. aeruginosa PAO1 genome revealed the presence of two open reading frames, mifS and mifR, which encodes putative TCS proteins, a histidine sensor kinase MifS and a response regulator MifR, respectively. This two-gene operon was found immediately upstream of the poxAB operon, where poxB encodes …

Identification Of Novel Cyclic Lipopeptides From A Positional Scanning Combinatorial Library With Enhanced Antibacterial And Antibiofilm Activities, Nina Bionda, Renee M. Fleeman, César De La Fuente-Núñez, Maria C. Rodriguez, Fany Reffuveille, Lindsey N. Shaw, Irena Pastar, Stephen C Davis, Robert E W Hancock, Predrag Cudic

Molecular Biosciences Faculty Publications

Treating bacterial infections can be difficult due to innate or acquired resistance mechanisms, and the formation of biofilms. Cyclic lipopeptides derived from fusaricidin/LI-F natural products represent particularly attractive candidates for the development of new antibacterial and antibiofilm agents, with the potential to meet the challenge of bacterial resistance to antibiotics. A positional-scanning combinatorial approach was used to identify the amino acid residues responsible for driving antibacterial activity, and increase the potency of these cyclic lipopeptides. Screening against the antibiotic resistant ESKAPE pathogens revealed the importance of hydrophobic as well as positively charged amino acid residues for activity of this class …

Characterization Of The Poxab Operon Encoding A Class D Carbapenemase In Pseudomonas Aeruginosa,, Diansy Zincke

FIU Electronic Theses and Dissertations

Pseudomonas aeruginosa is a dreaded opportunistic pathogen that causes severe and often intractable infections in immunocompromised and critically ill patients. This bacterium is also the primary cause of fatal lung infections in patients with cystic fibrosis and a leading nosocomial pathogen responsible for nearly 10% of all hospital-acquired infections. P. aeruginosa is intrinsically recalcitrant to most classes of antibiotics and has the ability to acquire additional resistance during treatment. In particular, resistance to the widely used β-lactam antibiotics is frequently mediated by the expression of AmpC, a chromosomally encoded β-lactamase that is ubiquitously found in P. aeruginosa strains. This dissertation …

An Experimental Study Of Strong Reciprocity In Bacteria, R. Inglis, R. Inglis, Stuart West, Angus Buckling

Biology Department Faculty Works

Strong reciprocity, whereby cooperators punish non-cooperators, may help to explain the evolutionary success of cooperative behaviours. However, theory suggests that selection for strong reciprocity can depend upon tight genetic linkage between cooperation and punishment, to avoid the strategy being outcompeted by non-punishing cooperators. We tested this hypothesis using experimental populations of the bacterium Pseudomonas aeruginosa, which cooperate by producing iron-scavenging siderophores and, in this context, punish non-cooperators with toxins. Consistent with theory, we show that cooperative punishers can indeed invade cheats, but only when the traits are tightly linked. These results emphasize that punishment is only likely to be favoured …

Opportunistic Pathogenic Bacteria Colonize Thorns Of Native Rio Grande Valley Plants, Felicia A. Charles, Sergio Cepeda, J. Andrew Mcdonald, Kristine L. Lowe

Biology Faculty Publications and Presentations

Thorns serve as mechanical defenses of plants against herbivory. However, plant thorns harbor microorganisms that are potentially pathogenic. These pathogens may be transferred to herbivores and other animals and provide an additional defense for the plants. Thorns from 5 plant species native to the Rio Grande Valley of South Texas were collected and used to isolate bacteria colonizing the thorn surface. Thorns, leaves and stems of plants were visualized using Scanning Electron Microscopy (SEM) to observe any bacteria on the plant surface. Isolated bacteria were tested for their ability to grow in aerobic versus anaerobic environments, to produce hemolysis, carbon …

Pseudomonas Aeruginosa Elastase Induces Il-8 Production In The Lung Cells Via The Epidermal Growth Factor/Extracellular Signal-Regulated Proteins/Nfîºb Pathway, Ali Azghani

Biology Faculty Publications and Presentations

Background: The induction of chemokine secretion by fibroblasts is crucial for the migration of leukocytes into the parenchyma of the injured lung. Several bacterial products activate the lung’s structural as well as immune cells to produce pro inflammatory cytokines and chemokines. We report that elastase from Pseudomonas aeruginosa (PE) evokes IL-8 mRNA expression and protein secretion in nonmalignant culture of human lung fibroblasts by activating the receptor for epidermal growth factor (EGFR) and downstream mitogen-activated protein kinases (MAPK) pathway. Methods: We utilized western blot analysis to detect phosphorylation of EGFR and signal transduction intermediates. Northern blot and ELISA analyses were …

Anti-Quorum Sensing Agents From South Florida Medicinal Plants And Their Attenuation Of Pseudomonas Aeruginosa Pathogenicity, Allison L. Adonizio

FIU Electronic Theses and Dissertations

With the difficulty in treating recalcitrant infections and the growing resistance to antibiotics, new therapeutic modalities are becoming increasingly necessary. The interruption of bacterial quorum sensing (QS), or cell-cell communication is known to attenuate virulence, while limiting selective pressure toward resistance. This study initiates an ethnobotanically-directed search for QS inhibiting agents in south Florida medicinal plants. Fifty plants were screened for anti-QS activity using two biomonitor strains, Chromobacterium violaceum and Agrobacterium tumefaciens. Of these plants, six showed QS inhibition: Conocarpus erectus L. (Combretaceae), Chamaecyce hypericifolia (L.) Millsp. (Euphorbiaceae), Callistemon viminalis (Sol.ex Gaertn.) G. Don (Myrtaceae), Bucida burceras L. (Combretaceae), Tetrazygia …

Mechanism Of Enhanced Activity Of Liposome-Entrapped Aminoglycosides Against Resistant Strains Of Pseudomonas Aeruginosa, Clement Mugabe, Majed Halwani, Ali Azghani, Robert M. Lafrenie, Abdelwahab Omri

Biology Faculty Publications and Presentations

Pseudomonas aeruginosa is inherently resistant to most conventional antibiotics. The mechanism of resistance of this bacterium is mainly associated with the low permeability of its outer membrane to these agents. We sought to assess the bactericidal efficacy of liposome-entrapped aminoglycosides against resistant clinical strains of P. aeruginosa and to define the mechanism of liposome-bacterium interactions. Aminoglycosides were incorporated into liposomes, and the bactericidal efficacies of both free and liposomal drugs were evaluated. To define the mechanism of liposome-bacterium interactions, transmission electron microscopy (TEM), flow cytometry, lipid mixing assay, and immunocytochemistry were employed. Encapsulation of aminoglycosides into liposomes significantly increased their …

Pyoverdine Production By Pseudomonas Aeruginosa Exposed To Metals Or An Oxidative Stress Agent, Kim-Hien Thi Dao, Katherine E. Hamer, Christine L. Clark, Lawrence G. Harshman

School of Biological Sciences: Faculty Publications

Siderophores are low molecular mass compounds used by many microorganisms to scavenge dissolved iron, which is typically rare in environments. Pseudomonas aeruginosa (PAO1) was exposed to metals and methyl viologen in low-iron medium in order to monitor the abundance of a specific siderophore (pyoverdine) and monitor growth over time. In this study it was discovered that cadmium can stimulate pyoverdine abundance. Cadmium may interact with and sequester the Fur regulatory protein, which represses siderophore synthesis under conditions of iron sufficiency. Mercury did not stimulate pyoverdine production at concentrations that strongly inhibited bacterial growth, suggesting that pyoverdine production is not governed …