{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T03:43:23Z","timestamp":1760240603081,"version":"build-2065373602"},"reference-count":40,"publisher":"MDPI AG","issue":"14","license":[{"start":{"date-parts":[[2019,7,23]],"date-time":"2019-07-23T00:00:00Z","timestamp":1563840000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"FCT\/MCTES through national funds","award":["UID\/QUI\/50006\/2019"],"award-info":[{"award-number":["UID\/QUI\/50006\/2019"]}]},{"name":"FAP-DF","award":["0193.000866\/2015"],"award-info":[{"award-number":["0193.000866\/2015"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Pseudomonas aeruginosa and Staphylococcus aureus are two major pathogens involved in a large variety of infections. Their co-occurrence in the same site of infection has been frequently reported and is linked to enhanced virulence and difficulty of treatment. Herein, the antimicrobial and antibiofilm activities of an intragenic antimicrobial peptide (IAP), named Hs02, which was uncovered from the human unconventional myosin 1H protein, were investigated against several P. aeruginosa and S. aureus strains, including multidrug-resistant (MDR) isolates. The antibiofilm activity was evaluated on single- and dual-species biofilms of P. aeruginosa and S. aureus. Moreover, the effect of peptide Hs02 on the membrane fluidity of the strains was assessed through Laurdan generalized polarization (GP). Minimum inhibitory concentration (MIC) values of peptide Hs02 ranged from 2 to 16 \u03bcg\/mL against all strains and MDR isolates. Though Hs02 was not able to hamper biofilm formation by some strains at sub-MIC values, it clearly affected 24 h preformed biofilms, especially by reducing the viability of the bacterial cells within the single- and dual-species biofilms, as shown by confocal laser scanning microscopy (CLSM) and atomic force microscopy (AFM) images. Laurdan GP values showed that Hs02 induces membrane rigidification in both P. aeruginosa and S. aureus. Peptide Hs02 can potentially be a lead for further improvement as an antibiofilm agent.<\/jats:p>","DOI":"10.3390\/ijms20143604","type":"journal-article","created":{"date-parts":[[2019,7,23]],"date-time":"2019-07-23T10:44:51Z","timestamp":1563878691000},"page":"3604","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":23,"title":["Intragenic Antimicrobial Peptide Hs02 Hampers the Proliferation of Single- and Dual-Species Biofilms of P. aeruginosa and S. aureus: A Promising Agent for Mitigation of Biofilm-Associated Infections"],"prefix":"10.3390","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8339-1964","authenticated-orcid":false,"given":"Lucinda J.","family":"Bessa","sequence":"first","affiliation":[{"name":"LAQV\/Requimte, Departamento de Qu\u00edmica e Bioqu\u00edmica, Faculdade de Ci\u00eancias da, Universidade do Porto, 4050-313 Porto, Portugal"}]},{"given":"Julia R.","family":"Manickchand","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de S\u00edntese e An\u00e1lise de Biomol\u00e9culas, Instituto de Qu\u00edmica, Universidade de Bras\u00edlia, UnB, Bras\u00edlia DF 70910-900, Brasil"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1821-1992","authenticated-orcid":false,"given":"Peter","family":"Eaton","sequence":"additional","affiliation":[{"name":"LAQV\/Requimte, Departamento de Qu\u00edmica e Bioqu\u00edmica, Faculdade de Ci\u00eancias da, Universidade do Porto, 4050-313 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1096-3236","authenticated-orcid":false,"given":"Jos\u00e9 Roberto S. A.","family":"Leite","sequence":"additional","affiliation":[{"name":"N\u00facleo de Pesquisa em Morfologia e Imunonologia Aplicada, NuPMIA, \u00c1rea de Morfologia, Faculdade de Medicina, FM, Universidade de Bras\u00edlia, UnB, Bras\u00edlia DF 70910-900, Brasil"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1615-0009","authenticated-orcid":false,"given":"Guilherme D.","family":"Brand","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de S\u00edntese e An\u00e1lise de Biomol\u00e9culas, Instituto de Qu\u00edmica, Universidade de Bras\u00edlia, UnB, Bras\u00edlia DF 70910-900, Brasil"}]},{"given":"Paula","family":"Gameiro","sequence":"additional","affiliation":[{"name":"LAQV\/Requimte, Departamento de Qu\u00edmica e Bioqu\u00edmica, Faculdade de Ci\u00eancias da, Universidade do Porto, 4050-313 Porto, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2019,7,23]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1038\/nrmicro.2015.8","article-title":"The biogeography of polymicrobial infection","volume":"14","author":"Stacy","year":"2016","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Kong, E.F., Tsui, C., Kuchar\u00edkov\u00e1, S., Andes, D., Van Dijck, P., and Jabra-Rizk, M.A. (2016). Commensal Protection of Staphylococcus aureus against Antimicrobials by Candida albicans Biofilm Matrix. mBio, 7.","DOI":"10.1128\/mBio.01365-16"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"363","DOI":"10.1080\/21505594.2017.1395129","article-title":"From the wound to the bench: Exoproteome interplay between wound-colonizing Staphylococcus aureus strains and co-existing bacteria","volume":"9","author":"Junker","year":"2018","journal-title":"Virulence"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"106","DOI":"10.3389\/fcimb.2017.00106","article-title":"In vivo and In vitro Interactions between Pseudomonas aeruginosa and Staphylococcus spp.","volume":"7","author":"Hotterbeekx","year":"2017","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"273","DOI":"10.12968\/jowc.2018.27.5.273","article-title":"Antibiotic resistance and biofilm tolerance: A combined threat in the treatment of chronic infections","volume":"27","author":"Bowler","year":"2018","journal-title":"J. Wound Care"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1016\/j.tim.2013.12.004","article-title":"Interactions in multispecies biofilms: Do they actually matter?","volume":"22","author":"Ren","year":"2014","journal-title":"Trends Microbiol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1111\/j.1469-0691.2012.04001.x","article-title":"The polymicrobial nature of biofilm infection","volume":"19","author":"Wolcott","year":"2013","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1111\/iwj.12049","article-title":"Bacterial isolates from infected wounds and their antibiotic susceptibility pattern: Some remarks about wound infection","volume":"12","author":"Bessa","year":"2015","journal-title":"Int. Wound J."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Radlinski, L., Rowe, S.E., Kartchner, L.B., Maile, R., Cairns, B.A., Vitko, N.P., Gode, C.J., Lachiewicz, A.M., Wolfgang, M.C., and Conlon, B.P. (2017). Pseudomonas aeruginosa exoproducts determine antibiotic efficacy against Staphylococcus aureus. PLoS Boil., 15.","DOI":"10.1371\/journal.pbio.2003981"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"281","DOI":"10.3389\/fphar.2018.00281","article-title":"Antimicrobial Peptides and Their Therapeutic Potential for Bacterial Skin Infections and Wounds","volume":"9","author":"Pfalzgraff","year":"2018","journal-title":"Front. Pharmacol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1016\/j.jpeds.2008.08.001","article-title":"Impact of Pseudomonas and Staphylococcus infection on inflammation and clinical status in young children with cystic fibrosis","volume":"154","author":"Sagel","year":"2009","journal-title":"J. Pediatr."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"497","DOI":"10.1016\/j.jcf.2012.12.003","article-title":"Association between Staphylococcus aureus alone or combined with Pseudomonas aeruginosa and the clinical condition of patients with cystic fibrosis","volume":"12","author":"Hubert","year":"2013","journal-title":"J. Cyst. Fibros."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Dalton, T., Dowd, S.E., Wolcott, R.D., Sun, Y., Watters, C., Griswold, J.A., and Rumbaugh, K.P. (2011). An in vivo polymicrobial biofilm wound infection model to study interspecies interactions. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0027317"},{"key":"ref_14","first-page":"128","article-title":"The role of multispecies social interactions in shaping Pseudomonas aeruginosa pathogenicity in the cystic fibrosis lung","volume":"364","author":"Fothergill","year":"2017","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"563","DOI":"10.1038\/nrmicro.2016.94","article-title":"Biofilms: An emergent form of bacterial life","volume":"14","author":"Flemming","year":"2016","journal-title":"Nat. Rev. Genet."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"740","DOI":"10.1038\/nrmicro.2017.99","article-title":"Targeting microbial biofilms: Current and prospective therapeutic strategies","volume":"15","author":"Koo","year":"2017","journal-title":"Nat. Rev. Genet."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1038\/s41522-017-0035-0","article-title":"Staphylococcus aureus interaction with Pseudomonas aeruginosa biofilm enhances tobramycin resistance","volume":"3","author":"Beaudoin","year":"2017","journal-title":"NPJ Biofilms Microbiomes"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1763","DOI":"10.1038\/s41598-018-19434-7","article-title":"Mechanisms driving the antibacterial and antibiofilm properties of Hp1404 and its analogue peptides against multidrug-resistant Pseudomonas aeruginosa","volume":"8","author":"Kim","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"405","DOI":"10.1016\/j.jmii.2016.12.005","article-title":"Antimicrobial peptides as potential anti-biofilm agents against multidrug-resistant bacteria","volume":"50","author":"Chung","year":"2017","journal-title":"J. Microbiol. Immunol. Infect."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"254","DOI":"10.1177\/0022034516679973","article-title":"Antimicrobial peptides: Mechanisms of action and resistance","volume":"96","author":"Bechinger","year":"2017","journal-title":"J. Dent. Res."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Kumar, P., Kizhakkedathu, J.N., and Straus, S.K. (2018). Antimicrobial Peptides: Diversity, Mechanism of Action and Strategies to Improve the Activity and Biocompatibility In Vivo. Biomolecules, 8.","DOI":"10.3390\/biom8010004"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1016\/j.mib.2016.05.016","article-title":"Anti-biofilm peptides as a new weapon in antimicrobial warfare","volume":"33","author":"Pletzer","year":"2016","journal-title":"Curr. Opin. Microbiol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1080\/08927014.2015.1021340","article-title":"BaAMPs: The database of biofilm-active antimicrobial peptides","volume":"31","author":"Maccari","year":"2015","journal-title":"Biofouling"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Brand, G.D., Magalh\u00e3es, M.T.Q., Tinoco, M.L.P., Arag\u00e3o, F.J.L., Nicoli, J., Kelly, S.M., Cooper, A., and Bloch, C. (2012). Probing Protein Sequences as Sources for Encrypted Antimicrobial Peptides. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0045848"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"13263","DOI":"10.1038\/s41598-017-13685-6","article-title":"Encrypted Antimicrobial Peptides from Plant Proteins","volume":"7","author":"Ramada","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1007\/978-1-60761-594-1_10","article-title":"Antimicrobial peptides: The LPS connection","volume":"618","author":"Giuliani","year":"2010","journal-title":"Methods Mol. Biol."},{"key":"ref_27","unstructured":"Brand, G.D., Ramada, M.H.S., Manickchand, J.R., Correa, R., Ribeiro, D.J.S., Michele, A., Santos, M.A., Vasconcelos, A.G., Abr\u00e3o, F.Y., and Prates, M.V. Intragenic antimicrobial peptides (IAPs) from human proteins with potent antimicrobial and anti-inflammatory activity. PLoS ONE, in press."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Monserrat-Martinez, A., Gambin, Y., and Sierecki, E. (2019). Thinking Outside the Bug: Molecular Targets and Strategies to Overcome Antibiotic Resistance. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20061255"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"521","DOI":"10.1016\/j.cis.2017.06.001","article-title":"The interaction of antimicrobial peptides with membranes","volume":"247","author":"Travkova","year":"2017","journal-title":"Adv. Colloid Interface Sci."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"150","DOI":"10.1016\/j.jphotobiol.2018.03.002","article-title":"Evaluation of membrane fluidity of multidrug-resistant isolates of Escherichia coli and Staphylococcus aureus in presence and absence of antibiotics","volume":"181","author":"Bessa","year":"2018","journal-title":"J. Photochem. Photobiol. B Boil."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"128","DOI":"10.1016\/j.dib.2018.09.106","article-title":"Data on Laurdan spectroscopic analyses to compare membrane fluidity between susceptible and multidrug-resistant bacteria","volume":"21","author":"Bessa","year":"2018","journal-title":"Data Brief"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1007\/978-1-4939-6634-9_10","article-title":"Measurement of cell membrane fluidity by Laurdan GP: Fluorescence spectroscopy and microscopy","volume":"1520","author":"Scheinpflug","year":"2017","journal-title":"Methods Mol. Biol."},{"key":"ref_33","first-page":"E7077","article-title":"Daptomycin inhibits cell envelope synthesis by interfering with fluid membrane microdomains","volume":"113","author":"Wenzel","year":"2016","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"717","DOI":"10.1039\/b416648p","article-title":"Natural products to drugs: Daptomycin and related lipopeptide antibiotics","volume":"22","author":"Baltz","year":"2005","journal-title":"Nat. Prod. Rep."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"5134","DOI":"10.1128\/AAC.00780-13","article-title":"Differential Adaptive Responses of Staphylococcus aureus to In Vitro Selection with Different Antimicrobial Peptides","volume":"57","author":"Shireen","year":"2013","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/j.foodres.2015.03.016","article-title":"Membrane fluidity-related adaptive response mechanisms of foodborne bacterial pathogens under environmental stresses","volume":"72","author":"Yoon","year":"2015","journal-title":"Food Res. Int."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1","DOI":"10.21769\/BioProtoc.3063","article-title":"Assessing Membrane Fluidity and Visualizing Fluid Membrane Domains in Bacteria Using Fluorescent Membrane Dyes","volume":"8","author":"Wenzel","year":"2018","journal-title":"Bio-Protoc."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Chan, W.C., and White, P. (2000). Basic procedures. Fmoc Solid Phase Peptide Synthesis: A Practical Approach, Oxford University Press.","DOI":"10.1093\/oso\/9780199637256.003.0007"},{"key":"ref_39","unstructured":"Clinical and Laboratory Standards Institute (CLSI) (2012). Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically, CLSI Standard M07. [11th ed.]."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"151","DOI":"10.2217\/fmb-2017-0175","article-title":"Synergistic and antibiofilm properties of ocellatin peptides against multidrug-resistant Pseudomonas aeruginosa","volume":"13","author":"Bessa","year":"2018","journal-title":"Futur. Microbiol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/20\/14\/3604\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T13:08:46Z","timestamp":1760188126000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/20\/14\/3604"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,7,23]]},"references-count":40,"journal-issue":{"issue":"14","published-online":{"date-parts":[[2019,7]]}},"alternative-id":["ijms20143604"],"URL":"https:\/\/doi.org\/10.3390\/ijms20143604","relation":{},"ISSN":["1422-0067"],"issn-type":[{"type":"electronic","value":"1422-0067"}],"subject":[],"published":{"date-parts":[[2019,7,23]]}}}