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Pubblicazioni Aperte DIgitali Sapienza > Scienze biochimiche "A. Rossi Fanelli" > BIOCHIMICA >

Please use this identifier to cite or link to this item: http://hdl.handle.net/10805/2415

Title: Membrane-active derivatives of the frog skin peptide Esculentin-1 against relevant human pathogens
Authors: LUCA, VINCENZO
Tutor: MANGONI, MARIA LUISA
BARRA, DONATELLA
Keywords: ANTIMICROBIAL PEPTIDE
ANTI-BIOFILM ACTIVITY
ANTI-CANDIDA ACTIVITY
CYSTIC FIBROSIS
MEMBRANE-PERMEABILIZATION
CAENORHABDITIS ELEGANS
SCANNING ELECTRON MICROSCOPY
SIMPLE INFECTION MODEL
ANTI-PSEUDOMONAS ACTIVITY
Issue Date: 3-Mar-2014
Abstract: Candida albicans represents one of the most prevalent species causing life-threatening fungal infections. Current treatments to defeat Candida albicans have become quite difficult, due to their toxic side effects and the emergence of resistant strains. Antimicrobial peptides (AMPs) are fascinating molecules with a potential role as novel anti-infective agents. However, only a few studies have been performed on their efficacy towards the most virulent hyphal phenotype of this pathogen. The purpose of this work is to evaluate the anti-Candida activity of the N-terminal 1–18 fragment of the frog skin AMP esculentin- 1b, Esc(1–18), under both in vitro and in vivo conditions using Caenorhabditis elegans as a simple host model for microbial infections. Our results demonstrate that Esc(1–18) caused a rapid reduction in the number of viable yeast cells and killing of the hyphal population. Esc(1–18)revealed a membrane perturbing effect which is likely the basis of its mode of action. Esc(1-18) is able (1) to kill both growing stages of Candida; (2) to promote survival of Candida-infected living organisms and (3) to inhibit transition of these fungal cells from the roundish yeast shape to the more dangerous hyphal form at sub-inhibitory concentrations. Pseudomonas aeruginosa is an opportunistic bacterial pathogen that forms sessile communities, named biofilms. The non-motile forms are very difficult to eradicate and are often associated with the establishment of persistent infections, especially in patients with cystic fibrosis. The resistance of P. aeruginosa to conventional antibiotics has become a growing health concern worldwide and has prompted the search for new anti-infective agents with new modes of action. Naturally occurring antimicrobial peptides (AMPs) represent promising future template candidates. Here we report on the potent activity and membrane-perturbing effects of the amphibian AMP esculentin(1-21), on both the free-living and sessile forms of P. aeruginosa, as a possible mechanism for biofilm disruption. Furthermore, the findings that esculentin(1-21) is able to prolong survival of animals in models of sepsis and pulmonary infection indicate that this peptide can be a promising template for the generation of new antibiotic formulations to advance care of infections caused by P. aeruginosa.
URI: http://hdl.handle.net/10805/2415
Research interests: Antimicrobial peptides, Cystic fibrosis, Candida infection, mechanism of action study of antimicrobial peptides.
Skills short description: 1)Microbiologia: isolamento e coltura di microrganismi patogeni e non, cinetiche di killing, MIC, saggi di sinergia, saggi dell’alone di inibizione, BIOFILM batterici e fungini, saggi di emolisi. Produzione di liposomi che mimano la membrana di eucarioti e batteri Gram-positivi e Gram-negativi. Capacità di lavorare in ambiente sterile (cappa biologica di II livello). Spettrofotometro, lettore micropiastre in fluorescenza, sonicatore, saggi ELISA; 2) Coltura di C. elegans e messa a punto di protocolli di infezione; 3) Utilizzo di microscopio ottico invertito in campo chiaro e fluorescenza; 4) Biologia Molecolare: disegno di primer, elettroforesi su gel di agarosio, colorazione argentica, co-immunoprecipitazione, colorazione comassie, SDS-PAGE, Western blot.
Personal skills keywords: Microbiology
Molecular Biology
Biochemistry
Optical microscope
Biofilm
Appears in PhD:BIOCHIMICA

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