In Vitro and In Vivo Evaluation of the De Novo Designed Antimicrobial Peptide P6.2 Against a KPC-Producing P. aeruginosa Clinical Isolate

Autores
Martínez, Melina María Belén; Corleto, Ingrid Merlina; Weschenfeller, Melanie Elizabeth; Urrea Montes, Santiago; Salomon, Camila Naiara; Gonzalez, Natalia; Garavaglia, Matías Javier; Faccone, Diego Francisco; Maffia, Paulo Cesar
Año de publicación
2025
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The antimicrobial peptide P6.2 was previously de novo designed as an alpha hélix cationic amphipathic molecule. In previous work, we have shown that this peptide displayed significant antimicrobial activity against both Gram-positive (Staphylococcus aureus) and Gram-negative (Pseudomonas aeruginosa) bacteria. However, while P6.2 lacked biofilminhibiting properties against the P. aeruginosa strain PA01, it displayed anti-inflammatory effects in a murine acute lung infection model challenged with this pathogen. In this work, the peptide P6.2 antimicrobial activity and its possible synergy with meropenem were evaluated both in vitro and in vivo using a Galleria mellonella infection model against a carbapenem-resistant KPC-producing clinical isolate of P. aeruginosa. Firstly, the cytotoxic effect of the peptide on A549 and RAW264.7 cell lines was assayed, showing no cytotoxicity at 64 μg/mL and below. Then, the MIC (minimal inhibitory concentration) and bactericidal effect against the carbapenemase-producing strain P. aeruginosa M13513 strain were determined. P6.2 showed a MIC between 32 and 64 μg/mL, and a rapid bactericidal activity against this strain (less than 45 min). The peptide stability at different temperaturas and in bovine serum at 37 ◦C was also analyzed, showing good stability and almost no degradation after 15 min of incubation at 100 ◦C or 24 h at 37 ◦C in serum, respectively. The antibiofilm activity was also evaluated, and although the peptide did not show biofilm inhibitory activity, it did demonstrate biofilm disruptive activity, together with bactericidal activity inside the pre-formed biofilm. The possible synergistic effect with the carbapenem meropenem was then analyzed in vitro by killing kinetics, revealing a synergistic interaction between P6.2 and the antibiotic against this strain. Finally, P6.2 was evaluated in vivo in the Galleria mellonella larvae infection model. Interestingly, in G. mellonella, P6.2 alone did not completely clear the infection caused by P. aeruginosa M13513. However, when combined with meropenem, P6.2 demonstrated a synergistic effect, leading to increased survival rates in infected larvae. The results presented here highlight the potential that this peptide displays when used in combination with carbapenems against a clinically relevant KPC-producing P. aeruginosa.
Fil: Martínez, Melina María Belén. Laboratorio de Biotecnologia y Microbiologia Aplicada ; Secretaria de Investigacion ; Universidad Nacional de Hurlingham; . Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Corleto, Ingrid Merlina. Laboratorio de Biotecnologia y Microbiologia Aplicada ; Secretaria de Investigacion ; Universidad Nacional de Hurlingham; . Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Weschenfeller, Melanie Elizabeth. Laboratorio de Biotecnologia y Microbiologia Aplicada ; Secretaria de Investigacion ; Universidad Nacional de Hurlingham; . Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Urrea Montes, Santiago. Laboratorio de Biotecnologia y Microbiologia Aplicada ; Secretaria de Investigacion ; Universidad Nacional de Hurlingham; . Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Salomon, Camila Naiara. Laboratorio de Biotecnologia y Microbiologia Aplicada ; Secretaria de Investigacion ; Universidad Nacional de Hurlingham; . Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas; Argentina
Fil: Gonzalez, Natalia. Laboratorio de Biotecnologia y Microbiologia Aplicada ; Secretaria de Investigacion ; Universidad Nacional de Hurlingham;
Fil: Garavaglia, Matías Javier. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Laboratorio de Biotecnologia y Microbiologia Aplicada ; Secretaria de Investigacion ; Universidad Nacional de Hurlingham;
Fil: Faccone, Diego Francisco. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas. Área de Antimicrobianos; Argentina
Fil: Maffia, Paulo Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas; Argentina
Materia
ANTIMICROBIAL
PEPTIDE
PSEUDOMONAS
AERUGINOSA
MEROPENEM
KPC
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/269383
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/269383
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling In Vitro and In Vivo Evaluation of the De Novo Designed Antimicrobial Peptide P6.2 Against a KPC-Producing P. aeruginosa Clinical IsolateMartínez, Melina María BelénCorleto, Ingrid MerlinaWeschenfeller, Melanie ElizabethUrrea Montes, SantiagoSalomon, Camila NaiaraGonzalez, NataliaGaravaglia, Matías JavierFaccone, Diego FranciscoMaffia, Paulo CesarANTIMICROBIALPEPTIDEPSEUDOMONASAERUGINOSAMEROPENEMKPChttps://purl.org/becyt/ford/3.4https://purl.org/becyt/ford/3The antimicrobial peptide P6.2 was previously de novo designed as an alpha hélix cationic amphipathic molecule. In previous work, we have shown that this peptide displayed significant antimicrobial activity against both Gram-positive (Staphylococcus aureus) and Gram-negative (Pseudomonas aeruginosa) bacteria. However, while P6.2 lacked biofilminhibiting properties against the P. aeruginosa strain PA01, it displayed anti-inflammatory effects in a murine acute lung infection model challenged with this pathogen. In this work, the peptide P6.2 antimicrobial activity and its possible synergy with meropenem were evaluated both in vitro and in vivo using a Galleria mellonella infection model against a carbapenem-resistant KPC-producing clinical isolate of P. aeruginosa. Firstly, the cytotoxic effect of the peptide on A549 and RAW264.7 cell lines was assayed, showing no cytotoxicity at 64 μg/mL and below. Then, the MIC (minimal inhibitory concentration) and bactericidal effect against the carbapenemase-producing strain P. aeruginosa M13513 strain were determined. P6.2 showed a MIC between 32 and 64 μg/mL, and a rapid bactericidal activity against this strain (less than 45 min). The peptide stability at different temperaturas and in bovine serum at 37 ◦C was also analyzed, showing good stability and almost no degradation after 15 min of incubation at 100 ◦C or 24 h at 37 ◦C in serum, respectively. The antibiofilm activity was also evaluated, and although the peptide did not show biofilm inhibitory activity, it did demonstrate biofilm disruptive activity, together with bactericidal activity inside the pre-formed biofilm. The possible synergistic effect with the carbapenem meropenem was then analyzed in vitro by killing kinetics, revealing a synergistic interaction between P6.2 and the antibiotic against this strain. Finally, P6.2 was evaluated in vivo in the Galleria mellonella larvae infection model. Interestingly, in G. mellonella, P6.2 alone did not completely clear the infection caused by P. aeruginosa M13513. However, when combined with meropenem, P6.2 demonstrated a synergistic effect, leading to increased survival rates in infected larvae. The results presented here highlight the potential that this peptide displays when used in combination with carbapenems against a clinically relevant KPC-producing P. aeruginosa.Fil: Martínez, Melina María Belén. Laboratorio de Biotecnologia y Microbiologia Aplicada ; Secretaria de Investigacion ; Universidad Nacional de Hurlingham; . Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Corleto, Ingrid Merlina. Laboratorio de Biotecnologia y Microbiologia Aplicada ; Secretaria de Investigacion ; Universidad Nacional de Hurlingham; . Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Weschenfeller, Melanie Elizabeth. Laboratorio de Biotecnologia y Microbiologia Aplicada ; Secretaria de Investigacion ; Universidad Nacional de Hurlingham; . Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Urrea Montes, Santiago. Laboratorio de Biotecnologia y Microbiologia Aplicada ; Secretaria de Investigacion ; Universidad Nacional de Hurlingham; . Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Salomon, Camila Naiara. Laboratorio de Biotecnologia y Microbiologia Aplicada ; Secretaria de Investigacion ; Universidad Nacional de Hurlingham; . Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas; ArgentinaFil: Gonzalez, Natalia. Laboratorio de Biotecnologia y Microbiologia Aplicada ; Secretaria de Investigacion ; Universidad Nacional de Hurlingham;Fil: Garavaglia, Matías Javier. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Laboratorio de Biotecnologia y Microbiologia Aplicada ; Secretaria de Investigacion ; Universidad Nacional de Hurlingham;Fil: Faccone, Diego Francisco. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas. Área de Antimicrobianos; ArgentinaFil: Maffia, Paulo Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas; ArgentinaMDPI2025-02info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/269383Martínez, Melina María Belén; Corleto, Ingrid Merlina; Weschenfeller, Melanie Elizabeth; Urrea Montes, Santiago; Salomon, Camila Naiara; et al.; In Vitro and In Vivo Evaluation of the De Novo Designed Antimicrobial Peptide P6.2 Against a KPC-Producing P. aeruginosa Clinical Isolate; MDPI; Biomolecules; 15; 3; 2-2025; 1-162218-273XCONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.mdpi.com/2218-273X/15/3/339info:eu-repo/semantics/altIdentifier/doi/10.3390/biom15030339info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:06:49Zoai:ri.conicet.gov.ar:11336/269383instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-03 10:06:50.136CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv In Vitro and In Vivo Evaluation of the De Novo Designed Antimicrobial Peptide P6.2 Against a KPC-Producing P. aeruginosa Clinical Isolate
title In Vitro and In Vivo Evaluation of the De Novo Designed Antimicrobial Peptide P6.2 Against a KPC-Producing P. aeruginosa Clinical Isolate
spellingShingle In Vitro and In Vivo Evaluation of the De Novo Designed Antimicrobial Peptide P6.2 Against a KPC-Producing P. aeruginosa Clinical Isolate
Martínez, Melina María Belén
ANTIMICROBIAL
PEPTIDE
PSEUDOMONAS
AERUGINOSA
MEROPENEM
KPC
title_short In Vitro and In Vivo Evaluation of the De Novo Designed Antimicrobial Peptide P6.2 Against a KPC-Producing P. aeruginosa Clinical Isolate
title_full In Vitro and In Vivo Evaluation of the De Novo Designed Antimicrobial Peptide P6.2 Against a KPC-Producing P. aeruginosa Clinical Isolate
title_fullStr In Vitro and In Vivo Evaluation of the De Novo Designed Antimicrobial Peptide P6.2 Against a KPC-Producing P. aeruginosa Clinical Isolate
title_full_unstemmed In Vitro and In Vivo Evaluation of the De Novo Designed Antimicrobial Peptide P6.2 Against a KPC-Producing P. aeruginosa Clinical Isolate
title_sort In Vitro and In Vivo Evaluation of the De Novo Designed Antimicrobial Peptide P6.2 Against a KPC-Producing P. aeruginosa Clinical Isolate
dc.creator.none.fl_str_mv Martínez, Melina María Belén
Corleto, Ingrid Merlina
Weschenfeller, Melanie Elizabeth
Urrea Montes, Santiago
Salomon, Camila Naiara
Gonzalez, Natalia
Garavaglia, Matías Javier
Faccone, Diego Francisco
Maffia, Paulo Cesar
author Martínez, Melina María Belén
author_facet Martínez, Melina María Belén
Corleto, Ingrid Merlina
Weschenfeller, Melanie Elizabeth
Urrea Montes, Santiago
Salomon, Camila Naiara
Gonzalez, Natalia
Garavaglia, Matías Javier
Faccone, Diego Francisco
Maffia, Paulo Cesar
author_role author
author2 Corleto, Ingrid Merlina
Weschenfeller, Melanie Elizabeth
Urrea Montes, Santiago
Salomon, Camila Naiara
Gonzalez, Natalia
Garavaglia, Matías Javier
Faccone, Diego Francisco
Maffia, Paulo Cesar
author2_role author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv ANTIMICROBIAL
PEPTIDE
PSEUDOMONAS
AERUGINOSA
MEROPENEM
KPC
topic ANTIMICROBIAL
PEPTIDE
PSEUDOMONAS
AERUGINOSA
MEROPENEM
KPC
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.4
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv The antimicrobial peptide P6.2 was previously de novo designed as an alpha hélix cationic amphipathic molecule. In previous work, we have shown that this peptide displayed significant antimicrobial activity against both Gram-positive (Staphylococcus aureus) and Gram-negative (Pseudomonas aeruginosa) bacteria. However, while P6.2 lacked biofilminhibiting properties against the P. aeruginosa strain PA01, it displayed anti-inflammatory effects in a murine acute lung infection model challenged with this pathogen. In this work, the peptide P6.2 antimicrobial activity and its possible synergy with meropenem were evaluated both in vitro and in vivo using a Galleria mellonella infection model against a carbapenem-resistant KPC-producing clinical isolate of P. aeruginosa. Firstly, the cytotoxic effect of the peptide on A549 and RAW264.7 cell lines was assayed, showing no cytotoxicity at 64 μg/mL and below. Then, the MIC (minimal inhibitory concentration) and bactericidal effect against the carbapenemase-producing strain P. aeruginosa M13513 strain were determined. P6.2 showed a MIC between 32 and 64 μg/mL, and a rapid bactericidal activity against this strain (less than 45 min). The peptide stability at different temperaturas and in bovine serum at 37 ◦C was also analyzed, showing good stability and almost no degradation after 15 min of incubation at 100 ◦C or 24 h at 37 ◦C in serum, respectively. The antibiofilm activity was also evaluated, and although the peptide did not show biofilm inhibitory activity, it did demonstrate biofilm disruptive activity, together with bactericidal activity inside the pre-formed biofilm. The possible synergistic effect with the carbapenem meropenem was then analyzed in vitro by killing kinetics, revealing a synergistic interaction between P6.2 and the antibiotic against this strain. Finally, P6.2 was evaluated in vivo in the Galleria mellonella larvae infection model. Interestingly, in G. mellonella, P6.2 alone did not completely clear the infection caused by P. aeruginosa M13513. However, when combined with meropenem, P6.2 demonstrated a synergistic effect, leading to increased survival rates in infected larvae. The results presented here highlight the potential that this peptide displays when used in combination with carbapenems against a clinically relevant KPC-producing P. aeruginosa.
Fil: Martínez, Melina María Belén. Laboratorio de Biotecnologia y Microbiologia Aplicada ; Secretaria de Investigacion ; Universidad Nacional de Hurlingham; . Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Corleto, Ingrid Merlina. Laboratorio de Biotecnologia y Microbiologia Aplicada ; Secretaria de Investigacion ; Universidad Nacional de Hurlingham; . Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Weschenfeller, Melanie Elizabeth. Laboratorio de Biotecnologia y Microbiologia Aplicada ; Secretaria de Investigacion ; Universidad Nacional de Hurlingham; . Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Urrea Montes, Santiago. Laboratorio de Biotecnologia y Microbiologia Aplicada ; Secretaria de Investigacion ; Universidad Nacional de Hurlingham; . Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Salomon, Camila Naiara. Laboratorio de Biotecnologia y Microbiologia Aplicada ; Secretaria de Investigacion ; Universidad Nacional de Hurlingham; . Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas; Argentina
Fil: Gonzalez, Natalia. Laboratorio de Biotecnologia y Microbiologia Aplicada ; Secretaria de Investigacion ; Universidad Nacional de Hurlingham;
Fil: Garavaglia, Matías Javier. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Laboratorio de Biotecnologia y Microbiologia Aplicada ; Secretaria de Investigacion ; Universidad Nacional de Hurlingham;
Fil: Faccone, Diego Francisco. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas. Área de Antimicrobianos; Argentina
Fil: Maffia, Paulo Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas; Argentina
description The antimicrobial peptide P6.2 was previously de novo designed as an alpha hélix cationic amphipathic molecule. In previous work, we have shown that this peptide displayed significant antimicrobial activity against both Gram-positive (Staphylococcus aureus) and Gram-negative (Pseudomonas aeruginosa) bacteria. However, while P6.2 lacked biofilminhibiting properties against the P. aeruginosa strain PA01, it displayed anti-inflammatory effects in a murine acute lung infection model challenged with this pathogen. In this work, the peptide P6.2 antimicrobial activity and its possible synergy with meropenem were evaluated both in vitro and in vivo using a Galleria mellonella infection model against a carbapenem-resistant KPC-producing clinical isolate of P. aeruginosa. Firstly, the cytotoxic effect of the peptide on A549 and RAW264.7 cell lines was assayed, showing no cytotoxicity at 64 μg/mL and below. Then, the MIC (minimal inhibitory concentration) and bactericidal effect against the carbapenemase-producing strain P. aeruginosa M13513 strain were determined. P6.2 showed a MIC between 32 and 64 μg/mL, and a rapid bactericidal activity against this strain (less than 45 min). The peptide stability at different temperaturas and in bovine serum at 37 ◦C was also analyzed, showing good stability and almost no degradation after 15 min of incubation at 100 ◦C or 24 h at 37 ◦C in serum, respectively. The antibiofilm activity was also evaluated, and although the peptide did not show biofilm inhibitory activity, it did demonstrate biofilm disruptive activity, together with bactericidal activity inside the pre-formed biofilm. The possible synergistic effect with the carbapenem meropenem was then analyzed in vitro by killing kinetics, revealing a synergistic interaction between P6.2 and the antibiotic against this strain. Finally, P6.2 was evaluated in vivo in the Galleria mellonella larvae infection model. Interestingly, in G. mellonella, P6.2 alone did not completely clear the infection caused by P. aeruginosa M13513. However, when combined with meropenem, P6.2 demonstrated a synergistic effect, leading to increased survival rates in infected larvae. The results presented here highlight the potential that this peptide displays when used in combination with carbapenems against a clinically relevant KPC-producing P. aeruginosa.
publishDate 2025
dc.date.none.fl_str_mv 2025-02
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/269383
Martínez, Melina María Belén; Corleto, Ingrid Merlina; Weschenfeller, Melanie Elizabeth; Urrea Montes, Santiago; Salomon, Camila Naiara; et al.; In Vitro and In Vivo Evaluation of the De Novo Designed Antimicrobial Peptide P6.2 Against a KPC-Producing P. aeruginosa Clinical Isolate; MDPI; Biomolecules; 15; 3; 2-2025; 1-16
2218-273X
CONICET Digital
CONICET
url http://hdl.handle.net/11336/269383
identifier_str_mv Martínez, Melina María Belén; Corleto, Ingrid Merlina; Weschenfeller, Melanie Elizabeth; Urrea Montes, Santiago; Salomon, Camila Naiara; et al.; In Vitro and In Vivo Evaluation of the De Novo Designed Antimicrobial Peptide P6.2 Against a KPC-Producing P. aeruginosa Clinical Isolate; MDPI; Biomolecules; 15; 3; 2-2025; 1-16
2218-273X
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.mdpi.com/2218-273X/15/3/339
info:eu-repo/semantics/altIdentifier/doi/10.3390/biom15030339
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
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application/pdf
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dc.publisher.none.fl_str_mv MDPI
publisher.none.fl_str_mv MDPI
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reponame_str CONICET Digital (CONICET)
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instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.13397

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