Caracterización del gen mucA en cepas de Pseudomonas Aeruginosa asociadas a la infección crónica producida en Fibrosis quística
Pseudomonas aeruginosa es un microorganismo de gran importancia a nivel clínico, se encuentra comúnmente aislado de infecciones asociadas a la atención en salud (IAAS) y también en pacientes con enfermedad de base como la Fibrosis Quística (F.Q). Dicha enfermedad genética es de tipo autosómico reces...
- Autores:
-
Vélez Lozano, Valeria Andrea
- Tipo de recurso:
- Trabajo de grado de pregrado
- Fecha de publicación:
- 2022
- Institución:
- Colegio Mayor de Cundinamarca
- Repositorio:
- Repositorio Colegio Mayor de Cundinamarca
- Idioma:
- spa
- OAI Identifier:
- oai:repositorio.unicolmayor.edu.co:unicolmayor/5730
- Acceso en línea:
- https://repositorio.unicolmayor.edu.co/handle/unicolmayor/5730
- Palabra clave:
- Aeruginosa
Fibrosis Quística
Gen mucA
- Rights
- closedAccess
- License
- Derechos Reservados - Universidad Colegio Mayor de Cundinamarca, 2022
id |
UCOLMAYOR2_5b630548abca7778e99ecc424a1744b5 |
---|---|
oai_identifier_str |
oai:repositorio.unicolmayor.edu.co:unicolmayor/5730 |
network_acronym_str |
UCOLMAYOR2 |
network_name_str |
Repositorio Colegio Mayor de Cundinamarca |
repository_id_str |
|
dc.title.spa.fl_str_mv |
Caracterización del gen mucA en cepas de Pseudomonas Aeruginosa asociadas a la infección crónica producida en Fibrosis quística |
title |
Caracterización del gen mucA en cepas de Pseudomonas Aeruginosa asociadas a la infección crónica producida en Fibrosis quística |
spellingShingle |
Caracterización del gen mucA en cepas de Pseudomonas Aeruginosa asociadas a la infección crónica producida en Fibrosis quística Aeruginosa Fibrosis Quística Gen mucA |
title_short |
Caracterización del gen mucA en cepas de Pseudomonas Aeruginosa asociadas a la infección crónica producida en Fibrosis quística |
title_full |
Caracterización del gen mucA en cepas de Pseudomonas Aeruginosa asociadas a la infección crónica producida en Fibrosis quística |
title_fullStr |
Caracterización del gen mucA en cepas de Pseudomonas Aeruginosa asociadas a la infección crónica producida en Fibrosis quística |
title_full_unstemmed |
Caracterización del gen mucA en cepas de Pseudomonas Aeruginosa asociadas a la infección crónica producida en Fibrosis quística |
title_sort |
Caracterización del gen mucA en cepas de Pseudomonas Aeruginosa asociadas a la infección crónica producida en Fibrosis quística |
dc.creator.fl_str_mv |
Vélez Lozano, Valeria Andrea |
dc.contributor.advisor.none.fl_str_mv |
Sánchez Mora, Ruth Mélida Romero Calderón, Ibeth Cristina |
dc.contributor.author.none.fl_str_mv |
Vélez Lozano, Valeria Andrea |
dc.subject.proposal.spa.fl_str_mv |
Aeruginosa Fibrosis Quística |
topic |
Aeruginosa Fibrosis Quística Gen mucA |
dc.subject.proposal.eng.fl_str_mv |
Gen mucA |
description |
Pseudomonas aeruginosa es un microorganismo de gran importancia a nivel clínico, se encuentra comúnmente aislado de infecciones asociadas a la atención en salud (IAAS) y también en pacientes con enfermedad de base como la Fibrosis Quística (F.Q). Dicha enfermedad genética es de tipo autosómico recesivo, caracterizada por tener una afectación multisistémica y, a su vez, por presentar acumulación de moco dentro de los pulmones de los pacientes, permitiendo la colonización de un amplio espectro de microorganismos. En el caso del estadio crónico de la infección producida por P. aeruginosa en los pacientes con F.Q, se presentan una gran variedad de mutaciones en el genoma, debido a la exposición prolongada a factores estresores, tales como el ambiente anaerobio producido por el moco, la insuficiencia nutricional, la presencia de altas concentraciones de antibióticos para erradicar la infección y la constante activación de la respuesta inmune encabezada por los neutrófilos y especies reactivas de oxígeno (ROS). Uno de los genes que se encuentra asociado al estadio crónico de la infección en pacientes con F.Q es el gen mucA, el cual en condiciones normales se encarga de controlar la producción del alginato. No obstante, cuando se presenta la mutación, se genera la conversión mucoide de la bacteria. En ese sentido, el objetivo de esta investigación es caracterizar el gen mucA en cepas de Pseudomonas aeruginosa que se encuentran asociadas a la infección crónica producida en fibrosis quística. |
publishDate |
2022 |
dc.date.accessioned.none.fl_str_mv |
2022-10-25T16:32:39Z |
dc.date.available.none.fl_str_mv |
2022-10-25T16:32:39Z |
dc.date.issued.none.fl_str_mv |
2022-04 |
dc.type.spa.fl_str_mv |
Trabajo de grado - Pregrado |
dc.type.coar.spa.fl_str_mv |
http://purl.org/coar/resource_type/c_7a1f |
dc.type.coarversion.spa.fl_str_mv |
http://purl.org/coar/version/c_970fb48d4fbd8a85 |
dc.type.content.spa.fl_str_mv |
Text |
dc.type.driver.spa.fl_str_mv |
info:eu-repo/semantics/bachelorThesis |
dc.type.redcol.spa.fl_str_mv |
https://purl.org/redcol/resource_type/TM |
dc.type.version.spa.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
http://purl.org/coar/resource_type/c_7a1f |
status_str |
publishedVersion |
dc.identifier.uri.none.fl_str_mv |
https://repositorio.unicolmayor.edu.co/handle/unicolmayor/5730 |
url |
https://repositorio.unicolmayor.edu.co/handle/unicolmayor/5730 |
dc.language.iso.spa.fl_str_mv |
spa |
language |
spa |
dc.relation.references.spa.fl_str_mv |
. Pachori P, Gothalwal R, Gandhi P. Emergence of antibiotic resistance Pseudomonas aeruginosa in intensive care unit; a critical review. Genes & Diseases [internet] 2019; 6 [cited 20 april 2021] Available in: https://www.sciencedirect.com/science/article/pii/S2352304219300170 Folkesson A, Jelsbak L, Yang L, Johansen HK, Ciofu O, Høiby N, Molin S. Adaptation of Pseudomonas aeruginosa to the cystic fibrosis airway: an evolutionary perspective. Nat Rev Microbiol [internet] 2012; 10 [cited 20 april 2021] Available in: https://pubmed.ncbi.nlm.nih.gov/23147702/ Sautter R, Ramos D, Schneper L, Ciofu O, Wassermann T, Koh CL, et al. A complex multilevel attack on Pseudomonas aeruginosa algT/U expression and AlgT/U activity results in the loss of alginate production. Gene [internet] 2012; 498 [cited 08 oct 2020] Available in: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4968699/ Jurado-Martín I, Sainz-Mejías M, McClean S. Pseudomonas aeruginosa: An Audacious Pathogen with an Adaptable Arsenal of Virulence Factors. International Journal of Molecular Sciences [internet] 2021; 22 [cited 20 april 2021] Available in: https://www.mdpi.com/1422-0067/22/6/3128#cite Gellatly SH, Hancock REW. Pseudomonas aeruginosa: new insights into pathogenesis and host defenses. Pathogens and Disease [internet] 2013; 67 [cited 02 jan 2021] Available in: https://academic.oup.com/femspd/article/67/3/159/2398791 Sol Z, Jiao X, Peng Q, Jiang F, Huang Y, Zhang J, et al. Antibiotic Resistance in Pseudomonas aeruginosa is Associated with Decreased Fitness. Cell Physiol Biochem [internet] 2013; 31 [cited 02 jan 2021] Available in: https://www.karger.com/Article/Pdf/343372 . Chang C. Surface Sensing for Biofilm Formation in Pseudomonas aeruginosa. Frontiers in Microbiology [internet] 2018; 8 [cited 17 jan 2021] Available in: https://www.frontiersin.org/articles/10.3389/fmicb.2017.02671/full Tacconelli E, Magrini N, Carmeli Y, Harbarth S, Kahlmeter G, Kluytmans J, et al. Global priority list of antibiotic-resistant bacteria to guide research, discovery, and development of new antibiotics. WHO [internet] 2017 [cited 17 jan 2021] Available in: https://www.who.int/medicines/publications/WHO-PPLShort_Summary_25Feb-ET_NM_WHO.pdf Pang Z, Raudonis R, Glick BR, Lin T, Cheng Z. Antibiotic resistance in Pseudomonas aeruginosa: mechanisms and alternative therapeutic strategies. Biotechnology Advances [internet] 2019; 37 [cited 17 jan 2021] Available in: https://www.sciencedirect.com/science/article/pii/S0734975018301976 Cystic Fibrosis Foundation. Patient Registry Annual Data Report to the Center Directors 2019. Cystic Fibrosis Foundation Patient Registry [internet] 2020 [cited 17 jan 2021] Available in: https://www.cff.org/Research/ResearcherResources/Patient-Registry/2019-Patient-Registry-Annual-Data-Report.pdf Bhagirath AY, Li Y, Somayajula Y, Dadashi M, Badr S, Duan K. Cystic fibrosis lung environment and Pseudomonas aeruginosa infection. BMC Pulmonary Medicine [internet] 2016; 16 [cited 17 jan 2021] Available in: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5139081/ anmanee W, Su S, Schurr MJ, Lau GW, Zhu X, et al. The anti-sigma factor MucA of Pseudomonas aeruginosa: Dramatic differences of a mucA22 vs. a ΔmucA mutant in anaerobic acidified nitrite sensitivity of planktonic and biofilm bacteria in vitro and during chronic murine lung infection. PLOS ONE [internet] 2019; 14 [cited 20 april 2021] Available in: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0216401 Vidya P, Smith L, Beaudoin T, Yau YCW, Clark S, Coburn B, et al. Chronic infection phenotypes of Pseudomonas aeruginosa are associated with failure of eradication in children with cystic fibrosis. EJCMID [internet] 2016; 35 [cited 17 jan 2021] Available in: https://link.springer.com/article/10.1007/s10096-015- 2509-4 Yin Y, Withers TR, Wang X, Yu WD. Evidence for Sigma Factor Competition in the Regulation of Alginate Production by Pseudomonas aeruginosa. PLOS ONE [internet] 2013; 8 [cited 17 jan 2021] Available in: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0072329 Cross AR, Raghuram V, Wang Z, Dey D, Goldberg JB. Overproduction of the AlgT Sigma Factor Is Lethal to Mucoid Pseudomonas aeruginosa. Journal of Bacteriology [internet] 2020; 202 [cited 20 april 2021] Available in: https://jb.asm.org/content/202/20/e00445-20 Feliziani S, Luján AM, Moyano AJ, Sola C, Bocco JL. Mucoidy, Quorum Sensing, Mismatch Repair and Antibiotic Resistance in Pseudomonas aeruginosa from Cystic Fibrosis Chronic Airways Infections. PLOS ONE [internet] 2010; 5 [cited 20 april 2021] Available in: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0012669 Pozuelo MJ, Jiménez PA, Valderrey AD, Fernández-Olmos A, Cantón R, Rotger R. Polimorfismo de los genes mucA y fpvA en Pseudomonas aeruginosa de pacientes con fibrosis quística: coexistencia de variantes genéticas en el mismo paciente. Enferm Infecc Microbiol Clin [internet] 2011; 29 [citado 20 abril 2021] Disponible en: https://www.elsevier.es/es-revistaenfermedades-infecciosas-microbiologia-clinica-28-pdf-S0213005X10004015 Candido N, Capizzani CP, Gomes LA, Marin M, Galetti R, Ciofu O, et al. Adaptation of Pseudomonas aeruginosa to the chronic phenotype by mutations in the algTmucABD operon in isolates from Brazilian cystic fibrosis patients. PLOS ONE [internet] 2018 [cited 17 jan 2021] Available in: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0208013 Schofield MC, Rodríguez D, Kidman AA, Cassin EK, Michaels LA, Campbell EA, et al. The anti-sigma factor MucA is required for viability in Pseudomonas aeruginosa. bIORxiv [internet] 2021 [cited 20 april 2021] Available in: https://www.biorxiv.org/content/10.1101/2020.08.17.253617v3.full Weihui W, Yongxin J, Fang B, Shouguang J. Chapter 41 - Pseudomonas aeruginosa. Molecular Medical Microbiology [internet] 2015; 2 [cited 20 april 2021] Available in: https://www.sciencedirect.com/science/article/pii/B978012397169200041X#! Winstanley C, O’Brien S, Brockhurst MA. Pseudomonas aeruginosa Evolutionary Adaptation and Diversification in Cystic Fibrosis Chronic Lung Infections. Trends in Microbiology [internet] 2016; 24 (5) [cited 19 jan 2021] Available in: https://www.cell.com/trends/microbiology/fulltext/S0966- 842X(16)00021- 4?_returnURL=https%3A%2F%2Flinkinghub.elsevier.com%2Fretrieve%2Fpii% 2FS0966842X16000214%3Fshowall%3Dtrue Azam MW, Khan AU. Updates on the pathogenicity status of Pseudomonas aeruginosa. Drug Discovery Today [internet] 2019; 24 (1) [cited 20 april 2021] Available in: https://www.sciencedirect.com/science/article/abs/pii/S1359644618302356 Su SS, Wynn KZ, Ngwe H. Isolation and Identification of Pseudomonas aeruginosa from the Clinical Soil. Research Gate [internet] 2018 [cited 19 jan 2021] Available in: https://www.researchgate.net/publication/335619903_Isolation_and_Identificati on_of_Pseudomonas_aeruginosa_from_the_Clinical_Soil Zheng P, Raudonis R, Glick BR, Tong-Jun L, Cheng Z. Antibiotic resistance in Pseudomonas aeruginosa: mechanisms and alternative therapeutic strategies. Biotechnology Advances [internet] 2019; 37 [cited 20 april 2021] Available in: https://www.sciencedirect.com/science/article/pii/S0734975018301976 European Centre for Disease. Prevention and Control Annual Epidemiological Report for 2017 Healthcare-associated infections in intensive care units. ECDC [internet] 2019 [cited 19 jan 2021] Available in: https://www.ecdc.europa.eu/sites/default/files/documents/AER_for_2017- HAI.pdf Tran MT, Wibowo D, Rehm BHA. Pseudomonas aeruginosa Biofilms. MDPI [internet] 2020; 21 [cited 19 jan 2021] Available in: https://www.mdpi.com/1422- 0067/21/22/8671/pdf Catellani C, Assael BM . Cystic fibrosis: a clinical view. Cell. Mol. Life Sci [internet] 2017; 74 [cited 19 jan 2021] Available in: https://link.springer.com/article/10.1007%2Fs00018-016-2393-9 Malhotra S, Hayes Jr D, Wozniak DJ. Cystic Fibrosis and Pseudomonas aeruginosa: the Host-Microbe Interface. Clinical Microbiology Reviews [internet] 2019; 32 [cited 19 jan 2021] Available in: https://cmr.asm.org/content/32/3/e00138-18 Brown S, White R, Tobin P. Keep them breathing. Cystic fibrosis pathophysiology, diagnosis, and treatment. AAPA [internet] 2017; 30 [cited 05 may 2021] Available in: https://journals.lww.com/jaapa/fulltext/2017/05000/keep_them_breathing__cysti c_fibrosis.4.aspx Farrell PM, White TB, Ren CL, Hempstead SE, Accurso F, Derichs N, et al. Diagnosis of Cystic Fibrosis: Consensus Guidelines from the Cystic Fibrosis Foundation. The Journal of Pediatrics [internet] 2017; 181 [cited 19 jan 2021] Available in: https://www.sciencedirect.com/science/article/pii/S0022347616310484 Hauser AR, Jain M, Bar-Meir M, McColley SA. Clinical Significance of Microbial Infection and Adaptation in Cystic Fibrosis. Clinical Microbiology Review [internet] 2011; 4 (1) [cited 19 jan 2021] Available in: https://cmr.asm.org/content/24/1/29 Jackson L, Waters V. Factors influencing the acquisition and eradication of early Pseudomonas aeruginosa infection in cystic fibrosis. Journal of Cystic Fibrosis [internet] 2020 [cited 19 jan 2021] Available in: https://scihub.se/https://doi.org/10.1016/j.jcf.2020.10.008 Taccetti G, Denton M, Hayes K, ECFS-CTN, Drevinek P, Sermet-Gaudelus I. A critical review of definitions used to describe Pseudomonas aeruginosa microbiological status in patients with cystic fibrosis for application in clinical trials. Journal of Cystic Fibrosis [internet] 2019; 19 [cited 19 jan 2021] Available in: https://www.cysticfibrosisjournal.com/article/S1569-1993(19)30867-7/fulltext Schick A, Kassen R. Rapid diversification of Pseudomonas aeruginosa in cystic fibrosis lung-like conditions. PNAS [internet] 2018; 115 (42) 19 [cited 19 jan 2021] Available in: https://www.pnas.org/content/115/42/10714.full Limoli DH, Rockel AB, Host KM, Jha A, Kopp BT, Hollis T, et al. Cationic Antimicrobial Peptides Promote Microbial Mutagenesis and Pathoadaptation in Chronic Infections. PLOS Pathogens [internet] 2014; 10 (4) [cited 19 jan 2021] Available in: https://journals.plos.org/plospathogens/article?id=10.1371/journal.ppat.1004083 Hay ID, Wang Y, Moradali MF, Rehman ZU, Rehm BHA. Genetics and regulation of bacterial alginate production. Environmental Microbiology [internet] 2014; 16 (10) [cited 19 jan 2021] Available in: https://sfamjournals.onlinelibrary.wiley.com/doi/full/10.1111/1462-2920.12389 Hatite WA, Abdulkadhim MS. Molecular Detection for Nosocomial Pseudomonas aeruginosa and its Relationship with multidrug Resistance, Isolated from Hospitals Environment. Medico-legal Update. [internet] 2020; 20 (1) [cited 06 may 2021] Available in: https://ijop.net/index.php/mlu/article/view/433 Moehario LH, Boestami HP, Edbert D, Tjoa E, Robertus T. Automation for the identification of Pseudomonas aeruginosa: Comparison of TDR-300B, VITEK®2 and VITEK®-MS. BioRxiv. [internet] 2019; [cited 06 may 2021] Available in: https://www.biorxiv.org/content/10.1101/510107v1.full El’Garch F, Jeannot K, Hocquet D, Llanes-Barakat C, Plésiat P. Cumulative Effects of Several Nonenzymatic Mechanisms on the Resistance of Pseudomonas aeruginosa to Aminoglycosides. Antimicrobial Agents and Chemotherapy [Internet] 2020; 51 [cited 28 jan 2022] Available in: https://journals.asm.org/doi/full/10.1128/AAC.00704-06 Cystic Fibrosis Foundation. Patient Registry Annual Data Report to the Center Directors 2020. Cystic Fibrosis Foundation Patient Registry [internet] 2021 [cited 28 jan 2022] Available in: https://www.cff.org/media/23476/download Scales BS, Erb-Downward JR, Huffnagle IM, LiPuma JJ, Huffnagle GB. Comparative genomics of Pseudomonas fluorescens subclade III strains from human lungs. BMC Genomics [Internet] 2015. [cited 28 jan 2022] Available in: https://bmcgenomics.biomedcentral.com/articles/10.1186/s12864-015-2261-2 Liu X, Cai J, Chen H, Zhong Q, Hou Y, et al. Antibacterial activity and mechanism of linalool against Pseudomonas aeruginosa. Microbial Pathogenesis [internet] 2020; 141 [cited 30 march 2022]. Available in: https://doi.org/10.1038/srep28063 Becker, L., Steglich, M., Fuchs, S. et al. Comparison of six commercial kits to extract bacterial chromosome and plasmid DNA for MiSeq sequencing. Sci Rep [internet] 2016;6 [cited 28 march 2022]. Available inhttps://doi.org/10.1038/srep28063 Candido N, da Costa CP, Montero LA, Oana RG, da Costa AL, et al. Adaptation of Pseudomonas aeruginosa to the chronic phenotype by mutations in the algTmucABD operon in isolates from Brazilian cystic fibrosis patients. PLOS ONE. [internet] 2018 [cited 29 march 2022]. Available in: https://doi.org/10.1371/journal.pone.0208013 Bragonzi A, Wiehlmann L, Klockgether J, Cramer N, Worlitzsch D, Doring G, et al. Sequence diversity of the mucABD locus in Pseudomonas aeruginosa isolates from patients with cystic fibrosis. Microbiology Society. [internet] 2006 [cited 30 march 2022] Available in: https://www.microbiologyresearch.org/content/journal/micro/10.1099/mic.0.2917 5-0;jsessionid=5cpIGMZ6xeYgWJqDxlHLVGw8.mbslive-10-240-10-188 |
dc.rights.spa.fl_str_mv |
Derechos Reservados - Universidad Colegio Mayor de Cundinamarca, 2022 |
dc.rights.uri.spa.fl_str_mv |
https://creativecommons.org/licenses/by-nc-sa/4.0/ |
dc.rights.accessrights.spa.fl_str_mv |
info:eu-repo/semantics/closedAccess |
dc.rights.creativecommons.spa.fl_str_mv |
Atribución-NoComercial-CompartirIgual 4.0 Internacional (CC BY-NC-SA 4.0) |
dc.rights.coar.spa.fl_str_mv |
http://purl.org/coar/access_right/c_14cb |
rights_invalid_str_mv |
Derechos Reservados - Universidad Colegio Mayor de Cundinamarca, 2022 https://creativecommons.org/licenses/by-nc-sa/4.0/ Atribución-NoComercial-CompartirIgual 4.0 Internacional (CC BY-NC-SA 4.0) http://purl.org/coar/access_right/c_14cb |
eu_rights_str_mv |
closedAccess |
dc.format.extent.spa.fl_str_mv |
54p. |
dc.format.mimetype.spa.fl_str_mv |
application/pdf |
dc.publisher.faculty.spa.fl_str_mv |
Facultad de Ciencias de la Salud |
dc.publisher.place.spa.fl_str_mv |
Bogotá |
dc.publisher.program.spa.fl_str_mv |
Bacteriología y Laboratorio Clínico |
institution |
Colegio Mayor de Cundinamarca |
bitstream.url.fl_str_mv |
https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5730/5/license.txt https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5730/1/Informe%20Final%20-%20Proyecto%20de%20Grado%20Valeria%20Velez.pdf https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5730/2/6.%20Diapositivas%20sustentaci%c3%b3n%20tesis%20pregrado%20-%20Valeria%20V%c3%a9lez.pdf https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5730/3/CARTA%20DERECHOS%20DE%20AUTOR%20Velez.pdf https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5730/4/4.%20FORMATO%20IDENTIFICACI%c3%93N%20TRABAJOS%20DE%20GRADO%20-%20Valeria%20Velez.docx.pdf https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5730/6/Informe%20Final%20-%20Proyecto%20de%20Grado%20Valeria%20Velez.pdf.txt https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5730/8/6.%20Diapositivas%20sustentaci%c3%b3n%20tesis%20pregrado%20-%20Valeria%20V%c3%a9lez.pdf.txt https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5730/10/CARTA%20DERECHOS%20DE%20AUTOR%20Velez.pdf.txt https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5730/12/4.%20FORMATO%20IDENTIFICACI%c3%93N%20TRABAJOS%20DE%20GRADO%20-%20Valeria%20Velez.docx.pdf.txt https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5730/7/Informe%20Final%20-%20Proyecto%20de%20Grado%20Valeria%20Velez.pdf.jpg https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5730/9/6.%20Diapositivas%20sustentaci%c3%b3n%20tesis%20pregrado%20-%20Valeria%20V%c3%a9lez.pdf.jpg https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5730/11/CARTA%20DERECHOS%20DE%20AUTOR%20Velez.pdf.jpg https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5730/13/4.%20FORMATO%20IDENTIFICACI%c3%93N%20TRABAJOS%20DE%20GRADO%20-%20Valeria%20Velez.docx.pdf.jpg |
bitstream.checksum.fl_str_mv |
2f9959eaf5b71fae44bbf9ec84150c7a 7c0b7703427d6c4655b6c1066ad43a98 57cc6f303de355837b9baec58b1a057b ce12287f76d8634687afd4306acad3d4 2f67983be5a12576dafb24385ad89f56 7ed1d2a7d1d41c0bf43ffb0186d58d7c c6ff442e10c61606648de79fb01c99dc 1a65c4666baf18e61d0f2b5543d309fc d92a97da0490a3c1eb911d5c8c7ead4c 3014fbea8e898386de04c9f3d0c6a4e5 82d58f1e8f69bdaf2fc055322f18d1d4 2291b2d523d454f5895e5add157bce33 4c348a6df0af64bd6ea786199dfc0077 |
bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 MD5 MD5 MD5 MD5 MD5 MD5 MD5 MD5 MD5 MD5 MD5 |
repository.name.fl_str_mv |
Biblioteca Digital Unicolmayor |
repository.mail.fl_str_mv |
repositorio@unicolmayor.edu.co |
_version_ |
1812210040059199488 |
spelling |
Sánchez Mora, Ruth Mélidaefe8f5fbfdf7ec7c24ba850634bfbb9aRomero Calderón, Ibeth Cristina1be6cd68f2562dc4b43f161eb1ab77a0600Vélez Lozano, Valeria Andrea67871a92b4e3f63a4e637be2cb9c422e2022-10-25T16:32:39Z2022-10-25T16:32:39Z2022-04https://repositorio.unicolmayor.edu.co/handle/unicolmayor/5730Pseudomonas aeruginosa es un microorganismo de gran importancia a nivel clínico, se encuentra comúnmente aislado de infecciones asociadas a la atención en salud (IAAS) y también en pacientes con enfermedad de base como la Fibrosis Quística (F.Q). Dicha enfermedad genética es de tipo autosómico recesivo, caracterizada por tener una afectación multisistémica y, a su vez, por presentar acumulación de moco dentro de los pulmones de los pacientes, permitiendo la colonización de un amplio espectro de microorganismos. En el caso del estadio crónico de la infección producida por P. aeruginosa en los pacientes con F.Q, se presentan una gran variedad de mutaciones en el genoma, debido a la exposición prolongada a factores estresores, tales como el ambiente anaerobio producido por el moco, la insuficiencia nutricional, la presencia de altas concentraciones de antibióticos para erradicar la infección y la constante activación de la respuesta inmune encabezada por los neutrófilos y especies reactivas de oxígeno (ROS). Uno de los genes que se encuentra asociado al estadio crónico de la infección en pacientes con F.Q es el gen mucA, el cual en condiciones normales se encarga de controlar la producción del alginato. No obstante, cuando se presenta la mutación, se genera la conversión mucoide de la bacteria. En ese sentido, el objetivo de esta investigación es caracterizar el gen mucA en cepas de Pseudomonas aeruginosa que se encuentran asociadas a la infección crónica producida en fibrosis quística.INTRODUCCIÓN. 1 Planteamiento del problema 1 Objetivo general .. 2 Objetivos específicos .. 2 Justificación 2 1. ANTECEDENTES 4 2. MARCO REFERENCIAL. 6 2.1 Generalidades microbiológicas de Pseudomonas aeruginosa.. 6 2.2 Importancia a nivel intrahospitalario y enfermedades asociadas a la bacteria . 7 2.2.1 Fibrosis Quística _7 2.2.1.1 Manifestaciones clínicas. 8 2.2.1.2 Diagnóstico . 8 2.3 Infección de Pseudomonas aeruginosa en pacientes en F.Q . 9 2.3.1 Etapas de la infección por P. aeruginosa _9 2.4 Regulación y biosíntesis del alginato. 10 3. DISEÑO METODOLÓGICO 12 3.1 Universo, población, muestra 12 Tipo de investigación_12 Nivel, alcance o enfoque de la investigación _12 Población objeto de estudio _12 Muestra 12 3.2 Técnicas y procedimientos.. 13 3.2.1 Conservación de la muestra clínica 13 3.2.2 Cultivo e identificación de Pseudomonas aeruginosa y Pseudomonas fluorescens _13 3.2.3 Curvas de crecimiento de los aislados clínicos y ATCC 27853 _13 3.2.4 Amplificación del gen mucA mediante Reacción en Cadena de la Polimerasa (PCR) 14 3.2.4.1 Extracción de ADN genómico a partir de los aislados clínicos y ATCC 27853 14 3.2.4.2 Cuantificación de ADN de los productos de extracción. 15 3.2.4.3 Búsqueda y comparación de las secuencias nucleotídicas del gen mucA mediante herramientas bioinformáticas 15 3.2.4.3.1 Alineamiento de la secuencia del gen mucA de las cepas Pseudomonas aeruginosa ATCC PAO1 y 27853 y Pseudomonas fluorescens SBW25. 16 3.2.4.4 Diseño de primers 16 3.2.4.5 Amplificación del gen mucA mediante la reacción en cadena de la polimerasa (PCR) 16 3.2.4.6 Electroforesis en gel de agarosa.. 17 3.2.4.7 Purificación y secuenciación del gen mucA 17 3.2.4.8 Análisis bioinformático de las secuencias obtenidas del gen mucA .. 17 4. RESULTADOS 19 4.1 Cultivo e identificación de aislados clínicos. 19 4.1.1 Perfil de resistencia de los aislados clínicos _20 4.2 Curvas de crecimiento de cepa ATCC 27853 y aislados clínicos .. 21 4.3 Aislamiento del gen mucA por PCR.. 23 4.3.1 Cuantificación y electroforesis del ADN genómico _23 4.3.2 Comprobación de primers forward y reverse in silico mediante herramientas bioinformáticas. _25 4.3.2.1 Alineamiento de las secuencias del gen mucA de las cepas Pseudomonas aeruginosa ATCC PAO1 y 27853 25 4.3.2.2 Alineamiento de secuencia del gen mucA de las cepas Pseudomonas aeruginosa ATCC 27853 y Pseudomonas fluorescens SBW25 . 26 4.3.3 Diseño de primers _27 4.3.4 Amplificación del gen mucA por medio de PCR _28 4.4 Purificación y secuenciamiento 29 5. DISCUSIÓN 35 6. CONCLUSIONES 37Pseudomonas aeruginosa es un microorganismo de gran importancia a nivel clínico, se encuentra comúnmente aislado de infecciones asociadas a la atención en salud (IAAS) y también en pacientes con enfermedad de base como la Fibrosis Quística (F.Q). Dicha enfermedad genética es de tipo autosómico recesivo, caracterizada por tener una afectación multisistémica y, a su vez, por presentar acumulación de moco dentro de los pulmones de los pacientes, permitiendo la colonización de un amplio espectro de microorganismos. En el caso del estadio crónico de la infección producida por P. aeruginosa en los pacientes con F.Q, se presentan una gran variedad de mutaciones en el genoma, debido a la exposición prolongada a factores estresores, tales como el ambiente anaerobio producido por el moco, la insuficiencia nutricional, la presencia de altas concentraciones de antibióticos para erradicar la infección y la constante activación de la respuesta inmune encabezada por los neutrófilos y especies reactivas de oxígeno (ROS). Uno de los genes que se encuentra asociado al estadio crónico de la infección en pacientes con F.Q es el gen mucA, el cual en condiciones normales se encarga de controlar la producción del alginato. No obstante, cuando se presenta la mutación, se genera la conversión mucoide de la bacteria. En ese sentido, el objetivo de esta investigación es caracterizar el gen mucA en cepas de Pseudomonas aeruginosa que se encuentran asociadas a la infección crónica producida en fibrosis quísticaPregradoBacteriólogo(a) y Laboratorista Clínico54p.application/pdfspaDerechos Reservados - Universidad Colegio Mayor de Cundinamarca, 2022https://creativecommons.org/licenses/by-nc-sa/4.0/info:eu-repo/semantics/closedAccessAtribución-NoComercial-CompartirIgual 4.0 Internacional (CC BY-NC-SA 4.0)http://purl.org/coar/access_right/c_14cbCaracterización del gen mucA en cepas de Pseudomonas Aeruginosa asociadas a la infección crónica producida en Fibrosis quísticaTrabajo de grado - Pregradohttp://purl.org/coar/resource_type/c_7a1fhttp://purl.org/coar/version/c_970fb48d4fbd8a85Textinfo:eu-repo/semantics/bachelorThesishttps://purl.org/redcol/resource_type/TMinfo:eu-repo/semantics/publishedVersionFacultad de Ciencias de la SaludBogotáBacteriología y Laboratorio Clínico. Pachori P, Gothalwal R, Gandhi P. Emergence of antibiotic resistance Pseudomonas aeruginosa in intensive care unit; a critical review. Genes & Diseases [internet] 2019; 6 [cited 20 april 2021] Available in: https://www.sciencedirect.com/science/article/pii/S2352304219300170Folkesson A, Jelsbak L, Yang L, Johansen HK, Ciofu O, Høiby N, Molin S. Adaptation of Pseudomonas aeruginosa to the cystic fibrosis airway: an evolutionary perspective. Nat Rev Microbiol [internet] 2012; 10 [cited 20 april 2021] Available in: https://pubmed.ncbi.nlm.nih.gov/23147702/Sautter R, Ramos D, Schneper L, Ciofu O, Wassermann T, Koh CL, et al. A complex multilevel attack on Pseudomonas aeruginosa algT/U expression and AlgT/U activity results in the loss of alginate production. Gene [internet] 2012; 498 [cited 08 oct 2020] Available in: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4968699/Jurado-Martín I, Sainz-Mejías M, McClean S. Pseudomonas aeruginosa: An Audacious Pathogen with an Adaptable Arsenal of Virulence Factors. International Journal of Molecular Sciences [internet] 2021; 22 [cited 20 april 2021] Available in: https://www.mdpi.com/1422-0067/22/6/3128#citeGellatly SH, Hancock REW. Pseudomonas aeruginosa: new insights into pathogenesis and host defenses. Pathogens and Disease [internet] 2013; 67 [cited 02 jan 2021] Available in: https://academic.oup.com/femspd/article/67/3/159/2398791Sol Z, Jiao X, Peng Q, Jiang F, Huang Y, Zhang J, et al. Antibiotic Resistance in Pseudomonas aeruginosa is Associated with Decreased Fitness. Cell Physiol Biochem [internet] 2013; 31 [cited 02 jan 2021] Available in: https://www.karger.com/Article/Pdf/343372. Chang C. Surface Sensing for Biofilm Formation in Pseudomonas aeruginosa. Frontiers in Microbiology [internet] 2018; 8 [cited 17 jan 2021] Available in: https://www.frontiersin.org/articles/10.3389/fmicb.2017.02671/fullTacconelli E, Magrini N, Carmeli Y, Harbarth S, Kahlmeter G, Kluytmans J, et al. Global priority list of antibiotic-resistant bacteria to guide research, discovery, and development of new antibiotics. WHO [internet] 2017 [cited 17 jan 2021] Available in: https://www.who.int/medicines/publications/WHO-PPLShort_Summary_25Feb-ET_NM_WHO.pdfPang Z, Raudonis R, Glick BR, Lin T, Cheng Z. Antibiotic resistance in Pseudomonas aeruginosa: mechanisms and alternative therapeutic strategies. Biotechnology Advances [internet] 2019; 37 [cited 17 jan 2021] Available in: https://www.sciencedirect.com/science/article/pii/S0734975018301976Cystic Fibrosis Foundation. Patient Registry Annual Data Report to the Center Directors 2019. Cystic Fibrosis Foundation Patient Registry [internet] 2020 [cited 17 jan 2021] Available in: https://www.cff.org/Research/ResearcherResources/Patient-Registry/2019-Patient-Registry-Annual-Data-Report.pdfBhagirath AY, Li Y, Somayajula Y, Dadashi M, Badr S, Duan K. Cystic fibrosis lung environment and Pseudomonas aeruginosa infection. BMC Pulmonary Medicine [internet] 2016; 16 [cited 17 jan 2021] Available in: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5139081/anmanee W, Su S, Schurr MJ, Lau GW, Zhu X, et al. The anti-sigma factor MucA of Pseudomonas aeruginosa: Dramatic differences of a mucA22 vs. a ΔmucA mutant in anaerobic acidified nitrite sensitivity of planktonic and biofilm bacteria in vitro and during chronic murine lung infection. PLOS ONE [internet] 2019; 14 [cited 20 april 2021] Available in: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0216401Vidya P, Smith L, Beaudoin T, Yau YCW, Clark S, Coburn B, et al. Chronic infection phenotypes of Pseudomonas aeruginosa are associated with failure of eradication in children with cystic fibrosis. EJCMID [internet] 2016; 35 [cited 17 jan 2021] Available in: https://link.springer.com/article/10.1007/s10096-015- 2509-4Yin Y, Withers TR, Wang X, Yu WD. Evidence for Sigma Factor Competition in the Regulation of Alginate Production by Pseudomonas aeruginosa. PLOS ONE [internet] 2013; 8 [cited 17 jan 2021] Available in: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0072329Cross AR, Raghuram V, Wang Z, Dey D, Goldberg JB. Overproduction of the AlgT Sigma Factor Is Lethal to Mucoid Pseudomonas aeruginosa. Journal of Bacteriology [internet] 2020; 202 [cited 20 april 2021] Available in: https://jb.asm.org/content/202/20/e00445-20Feliziani S, Luján AM, Moyano AJ, Sola C, Bocco JL. Mucoidy, Quorum Sensing, Mismatch Repair and Antibiotic Resistance in Pseudomonas aeruginosa from Cystic Fibrosis Chronic Airways Infections. PLOS ONE [internet] 2010; 5 [cited 20 april 2021] Available in: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0012669Pozuelo MJ, Jiménez PA, Valderrey AD, Fernández-Olmos A, Cantón R, Rotger R. Polimorfismo de los genes mucA y fpvA en Pseudomonas aeruginosa de pacientes con fibrosis quística: coexistencia de variantes genéticas en el mismo paciente. Enferm Infecc Microbiol Clin [internet] 2011; 29 [citado 20 abril 2021] Disponible en: https://www.elsevier.es/es-revistaenfermedades-infecciosas-microbiologia-clinica-28-pdf-S0213005X10004015Candido N, Capizzani CP, Gomes LA, Marin M, Galetti R, Ciofu O, et al. Adaptation of Pseudomonas aeruginosa to the chronic phenotype by mutations in the algTmucABD operon in isolates from Brazilian cystic fibrosis patients. PLOS ONE [internet] 2018 [cited 17 jan 2021] Available in: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0208013Schofield MC, Rodríguez D, Kidman AA, Cassin EK, Michaels LA, Campbell EA, et al. The anti-sigma factor MucA is required for viability in Pseudomonas aeruginosa. bIORxiv [internet] 2021 [cited 20 april 2021] Available in: https://www.biorxiv.org/content/10.1101/2020.08.17.253617v3.fullWeihui W, Yongxin J, Fang B, Shouguang J. Chapter 41 - Pseudomonas aeruginosa. Molecular Medical Microbiology [internet] 2015; 2 [cited 20 april 2021] Available in: https://www.sciencedirect.com/science/article/pii/B978012397169200041X#!Winstanley C, O’Brien S, Brockhurst MA. Pseudomonas aeruginosa Evolutionary Adaptation and Diversification in Cystic Fibrosis Chronic Lung Infections. Trends in Microbiology [internet] 2016; 24 (5) [cited 19 jan 2021] Available in: https://www.cell.com/trends/microbiology/fulltext/S0966- 842X(16)00021- 4?_returnURL=https%3A%2F%2Flinkinghub.elsevier.com%2Fretrieve%2Fpii% 2FS0966842X16000214%3Fshowall%3DtrueAzam MW, Khan AU. Updates on the pathogenicity status of Pseudomonas aeruginosa. Drug Discovery Today [internet] 2019; 24 (1) [cited 20 april 2021] Available in: https://www.sciencedirect.com/science/article/abs/pii/S1359644618302356Su SS, Wynn KZ, Ngwe H. Isolation and Identification of Pseudomonas aeruginosa from the Clinical Soil. Research Gate [internet] 2018 [cited 19 jan 2021] Available in: https://www.researchgate.net/publication/335619903_Isolation_and_Identificati on_of_Pseudomonas_aeruginosa_from_the_Clinical_SoilZheng P, Raudonis R, Glick BR, Tong-Jun L, Cheng Z. Antibiotic resistance in Pseudomonas aeruginosa: mechanisms and alternative therapeutic strategies. Biotechnology Advances [internet] 2019; 37 [cited 20 april 2021] Available in: https://www.sciencedirect.com/science/article/pii/S0734975018301976European Centre for Disease. Prevention and Control Annual Epidemiological Report for 2017 Healthcare-associated infections in intensive care units. ECDC [internet] 2019 [cited 19 jan 2021] Available in: https://www.ecdc.europa.eu/sites/default/files/documents/AER_for_2017- HAI.pdfTran MT, Wibowo D, Rehm BHA. Pseudomonas aeruginosa Biofilms. MDPI [internet] 2020; 21 [cited 19 jan 2021] Available in: https://www.mdpi.com/1422- 0067/21/22/8671/pdfCatellani C, Assael BM . Cystic fibrosis: a clinical view. Cell. Mol. Life Sci [internet] 2017; 74 [cited 19 jan 2021] Available in: https://link.springer.com/article/10.1007%2Fs00018-016-2393-9Malhotra S, Hayes Jr D, Wozniak DJ. Cystic Fibrosis and Pseudomonas aeruginosa: the Host-Microbe Interface. Clinical Microbiology Reviews [internet] 2019; 32 [cited 19 jan 2021] Available in: https://cmr.asm.org/content/32/3/e00138-18Brown S, White R, Tobin P. Keep them breathing. Cystic fibrosis pathophysiology, diagnosis, and treatment. AAPA [internet] 2017; 30 [cited 05 may 2021] Available in: https://journals.lww.com/jaapa/fulltext/2017/05000/keep_them_breathing__cysti c_fibrosis.4.aspxFarrell PM, White TB, Ren CL, Hempstead SE, Accurso F, Derichs N, et al. Diagnosis of Cystic Fibrosis: Consensus Guidelines from the Cystic Fibrosis Foundation. The Journal of Pediatrics [internet] 2017; 181 [cited 19 jan 2021] Available in: https://www.sciencedirect.com/science/article/pii/S0022347616310484Hauser AR, Jain M, Bar-Meir M, McColley SA. Clinical Significance of Microbial Infection and Adaptation in Cystic Fibrosis. Clinical Microbiology Review [internet] 2011; 4 (1) [cited 19 jan 2021] Available in: https://cmr.asm.org/content/24/1/29Jackson L, Waters V. Factors influencing the acquisition and eradication of early Pseudomonas aeruginosa infection in cystic fibrosis. Journal of Cystic Fibrosis [internet] 2020 [cited 19 jan 2021] Available in: https://scihub.se/https://doi.org/10.1016/j.jcf.2020.10.008Taccetti G, Denton M, Hayes K, ECFS-CTN, Drevinek P, Sermet-Gaudelus I. A critical review of definitions used to describe Pseudomonas aeruginosa microbiological status in patients with cystic fibrosis for application in clinical trials. Journal of Cystic Fibrosis [internet] 2019; 19 [cited 19 jan 2021] Available in: https://www.cysticfibrosisjournal.com/article/S1569-1993(19)30867-7/fulltextSchick A, Kassen R. Rapid diversification of Pseudomonas aeruginosa in cystic fibrosis lung-like conditions. PNAS [internet] 2018; 115 (42) 19 [cited 19 jan 2021] Available in: https://www.pnas.org/content/115/42/10714.fullLimoli DH, Rockel AB, Host KM, Jha A, Kopp BT, Hollis T, et al. Cationic Antimicrobial Peptides Promote Microbial Mutagenesis and Pathoadaptation in Chronic Infections. PLOS Pathogens [internet] 2014; 10 (4) [cited 19 jan 2021] Available in: https://journals.plos.org/plospathogens/article?id=10.1371/journal.ppat.1004083Hay ID, Wang Y, Moradali MF, Rehman ZU, Rehm BHA. Genetics and regulation of bacterial alginate production. Environmental Microbiology [internet] 2014; 16 (10) [cited 19 jan 2021] Available in: https://sfamjournals.onlinelibrary.wiley.com/doi/full/10.1111/1462-2920.12389Hatite WA, Abdulkadhim MS. Molecular Detection for Nosocomial Pseudomonas aeruginosa and its Relationship with multidrug Resistance, Isolated from Hospitals Environment. Medico-legal Update. [internet] 2020; 20 (1) [cited 06 may 2021] Available in: https://ijop.net/index.php/mlu/article/view/433Moehario LH, Boestami HP, Edbert D, Tjoa E, Robertus T. Automation for the identification of Pseudomonas aeruginosa: Comparison of TDR-300B, VITEK®2 and VITEK®-MS. BioRxiv. [internet] 2019; [cited 06 may 2021] Available in: https://www.biorxiv.org/content/10.1101/510107v1.fullEl’Garch F, Jeannot K, Hocquet D, Llanes-Barakat C, Plésiat P. Cumulative Effects of Several Nonenzymatic Mechanisms on the Resistance of Pseudomonas aeruginosa to Aminoglycosides. Antimicrobial Agents and Chemotherapy [Internet] 2020; 51 [cited 28 jan 2022] Available in: https://journals.asm.org/doi/full/10.1128/AAC.00704-06Cystic Fibrosis Foundation. Patient Registry Annual Data Report to the Center Directors 2020. Cystic Fibrosis Foundation Patient Registry [internet] 2021 [cited 28 jan 2022] Available in: https://www.cff.org/media/23476/downloadScales BS, Erb-Downward JR, Huffnagle IM, LiPuma JJ, Huffnagle GB. Comparative genomics of Pseudomonas fluorescens subclade III strains from human lungs. BMC Genomics [Internet] 2015. [cited 28 jan 2022] Available in: https://bmcgenomics.biomedcentral.com/articles/10.1186/s12864-015-2261-2Liu X, Cai J, Chen H, Zhong Q, Hou Y, et al. Antibacterial activity and mechanism of linalool against Pseudomonas aeruginosa. Microbial Pathogenesis [internet] 2020; 141 [cited 30 march 2022]. Available in: https://doi.org/10.1038/srep28063Becker, L., Steglich, M., Fuchs, S. et al. Comparison of six commercial kits to extract bacterial chromosome and plasmid DNA for MiSeq sequencing. Sci Rep [internet] 2016;6 [cited 28 march 2022]. Available inhttps://doi.org/10.1038/srep28063Candido N, da Costa CP, Montero LA, Oana RG, da Costa AL, et al. Adaptation of Pseudomonas aeruginosa to the chronic phenotype by mutations in the algTmucABD operon in isolates from Brazilian cystic fibrosis patients. PLOS ONE. [internet] 2018 [cited 29 march 2022]. Available in: https://doi.org/10.1371/journal.pone.0208013Bragonzi A, Wiehlmann L, Klockgether J, Cramer N, Worlitzsch D, Doring G, et al. Sequence diversity of the mucABD locus in Pseudomonas aeruginosa isolates from patients with cystic fibrosis. Microbiology Society. [internet] 2006 [cited 30 march 2022] Available in: https://www.microbiologyresearch.org/content/journal/micro/10.1099/mic.0.2917 5-0;jsessionid=5cpIGMZ6xeYgWJqDxlHLVGw8.mbslive-10-240-10-188AeruginosaFibrosis QuísticaGen mucALICENSElicense.txtlicense.txttext/plain; charset=utf-814828https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5730/5/license.txt2f9959eaf5b71fae44bbf9ec84150c7aMD55open accessORIGINALInforme Final - Proyecto de Grado Valeria Velez.pdfInforme Final - Proyecto de Grado Valeria Velez.pdfapplication/pdf2832150https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5730/1/Informe%20Final%20-%20Proyecto%20de%20Grado%20Valeria%20Velez.pdf7c0b7703427d6c4655b6c1066ad43a98MD51open access6. Diapositivas sustentación tesis pregrado - Valeria Vélez.pdf6. Diapositivas sustentación tesis pregrado - Valeria Vélez.pdfapplication/pdf2852354https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5730/2/6.%20Diapositivas%20sustentaci%c3%b3n%20tesis%20pregrado%20-%20Valeria%20V%c3%a9lez.pdf57cc6f303de355837b9baec58b1a057bMD52open accessCARTA DERECHOS DE AUTOR Velez.pdfCARTA DERECHOS DE AUTOR Velez.pdfapplication/pdf154016https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5730/3/CARTA%20DERECHOS%20DE%20AUTOR%20Velez.pdfce12287f76d8634687afd4306acad3d4MD53metadata only access4. FORMATO IDENTIFICACIÓN TRABAJOS DE GRADO - Valeria Velez.docx.pdf4. FORMATO IDENTIFICACIÓN TRABAJOS DE GRADO - Valeria Velez.docx.pdfapplication/pdf132840https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5730/4/4.%20FORMATO%20IDENTIFICACI%c3%93N%20TRABAJOS%20DE%20GRADO%20-%20Valeria%20Velez.docx.pdf2f67983be5a12576dafb24385ad89f56MD54metadata only accessTEXTInforme Final - Proyecto de Grado Valeria Velez.pdf.txtInforme Final - Proyecto de Grado Valeria Velez.pdf.txtExtracted texttext/plain79197https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5730/6/Informe%20Final%20-%20Proyecto%20de%20Grado%20Valeria%20Velez.pdf.txt7ed1d2a7d1d41c0bf43ffb0186d58d7cMD56open access6. Diapositivas sustentación tesis pregrado - Valeria Vélez.pdf.txt6. Diapositivas sustentación tesis pregrado - Valeria Vélez.pdf.txtExtracted texttext/plain13133https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5730/8/6.%20Diapositivas%20sustentaci%c3%b3n%20tesis%20pregrado%20-%20Valeria%20V%c3%a9lez.pdf.txtc6ff442e10c61606648de79fb01c99dcMD58open accessCARTA DERECHOS DE AUTOR Velez.pdf.txtCARTA DERECHOS DE AUTOR Velez.pdf.txtExtracted texttext/plain1126https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5730/10/CARTA%20DERECHOS%20DE%20AUTOR%20Velez.pdf.txt1a65c4666baf18e61d0f2b5543d309fcMD510metadata only access4. FORMATO IDENTIFICACIÓN TRABAJOS DE GRADO - Valeria Velez.docx.pdf.txt4. FORMATO IDENTIFICACIÓN TRABAJOS DE GRADO - Valeria Velez.docx.pdf.txtExtracted texttext/plain3075https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5730/12/4.%20FORMATO%20IDENTIFICACI%c3%93N%20TRABAJOS%20DE%20GRADO%20-%20Valeria%20Velez.docx.pdf.txtd92a97da0490a3c1eb911d5c8c7ead4cMD512metadata only accessTHUMBNAILInforme Final - Proyecto de Grado Valeria Velez.pdf.jpgInforme Final - Proyecto de Grado Valeria Velez.pdf.jpgGenerated Thumbnailimage/jpeg7149https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5730/7/Informe%20Final%20-%20Proyecto%20de%20Grado%20Valeria%20Velez.pdf.jpg3014fbea8e898386de04c9f3d0c6a4e5MD57open access6. Diapositivas sustentación tesis pregrado - Valeria Vélez.pdf.jpg6. Diapositivas sustentación tesis pregrado - Valeria Vélez.pdf.jpgGenerated Thumbnailimage/jpeg5916https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5730/9/6.%20Diapositivas%20sustentaci%c3%b3n%20tesis%20pregrado%20-%20Valeria%20V%c3%a9lez.pdf.jpg82d58f1e8f69bdaf2fc055322f18d1d4MD59open accessCARTA DERECHOS DE AUTOR Velez.pdf.jpgCARTA DERECHOS DE AUTOR Velez.pdf.jpgGenerated Thumbnailimage/jpeg9319https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5730/11/CARTA%20DERECHOS%20DE%20AUTOR%20Velez.pdf.jpg2291b2d523d454f5895e5add157bce33MD511metadata only access4. FORMATO IDENTIFICACIÓN TRABAJOS DE GRADO - Valeria Velez.docx.pdf.jpg4. FORMATO IDENTIFICACIÓN TRABAJOS DE GRADO - Valeria Velez.docx.pdf.jpgGenerated Thumbnailimage/jpeg13995https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5730/13/4.%20FORMATO%20IDENTIFICACI%c3%93N%20TRABAJOS%20DE%20GRADO%20-%20Valeria%20Velez.docx.pdf.jpg4c348a6df0af64bd6ea786199dfc0077MD513metadata only accessunicolmayor/5730oai:repositorio.unicolmayor.edu.co:unicolmayor/57302024-05-10 12:31:47.141An error occurred on the license name.|||https://creativecommons.org/licenses/by-nc-sa/4.0/open accessBiblioteca Digital Unicolmayorrepositorio@unicolmayor.edu.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 |