Identificación de polimorfismos en el gen Mieloperoxidasa y su papel en Candidiasis Vulvovaginal recurrente por Candida SPP en pacientes de Bogotá DC
La mieloperoxidasa (MPO) es una proteína antimicrobiana presente en los gránulos azurófilos de los neutrófilos, su acción recae cuando se conjuga en el fagosoma con peróxido de hidrógeno, resultando en sustancias tóxicas para los microorganismos fagocitados (1). Se ha descrito que las infecciones vu...
- Autores:
-
Montoya Estrada, María Paula
Rincón González, Rincón González
- 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/5717
- Acceso en línea:
- https://repositorio.unicolmayor.edu.co/handle/unicolmayor/5717
- Palabra clave:
- Polimorfismos
Deficiencia de Mieloperoxidasa
Análisis Bioinformático
PCR
Secuenciación
Candidiasis vulvovaginal
Candida spp
- Rights
- closedAccess
- License
- Derechos Reservados - Universidad Colegio Mayor de Cundinamarca, 2022
| id |
UCOLMAYOR2_289cdeb66718b9723858fb52be7b5924 |
|---|---|
| oai_identifier_str |
oai:repositorio.unicolmayor.edu.co:unicolmayor/5717 |
| network_acronym_str |
UCOLMAYOR2 |
| network_name_str |
Repositorio Colegio Mayor de Cundinamarca |
| repository_id_str |
|
| dc.title.spa.fl_str_mv |
Identificación de polimorfismos en el gen Mieloperoxidasa y su papel en Candidiasis Vulvovaginal recurrente por Candida SPP en pacientes de Bogotá DC |
| title |
Identificación de polimorfismos en el gen Mieloperoxidasa y su papel en Candidiasis Vulvovaginal recurrente por Candida SPP en pacientes de Bogotá DC |
| spellingShingle |
Identificación de polimorfismos en el gen Mieloperoxidasa y su papel en Candidiasis Vulvovaginal recurrente por Candida SPP en pacientes de Bogotá DC Polimorfismos Deficiencia de Mieloperoxidasa Análisis Bioinformático PCR Secuenciación Candidiasis vulvovaginal Candida spp |
| title_short |
Identificación de polimorfismos en el gen Mieloperoxidasa y su papel en Candidiasis Vulvovaginal recurrente por Candida SPP en pacientes de Bogotá DC |
| title_full |
Identificación de polimorfismos en el gen Mieloperoxidasa y su papel en Candidiasis Vulvovaginal recurrente por Candida SPP en pacientes de Bogotá DC |
| title_fullStr |
Identificación de polimorfismos en el gen Mieloperoxidasa y su papel en Candidiasis Vulvovaginal recurrente por Candida SPP en pacientes de Bogotá DC |
| title_full_unstemmed |
Identificación de polimorfismos en el gen Mieloperoxidasa y su papel en Candidiasis Vulvovaginal recurrente por Candida SPP en pacientes de Bogotá DC |
| title_sort |
Identificación de polimorfismos en el gen Mieloperoxidasa y su papel en Candidiasis Vulvovaginal recurrente por Candida SPP en pacientes de Bogotá DC |
| dc.creator.fl_str_mv |
Montoya Estrada, María Paula Rincón González, Rincón González |
| dc.contributor.advisor.none.fl_str_mv |
Cruz Baquero, Claudia Andrea Pinilla Bermúdez, Gladys |
| dc.contributor.author.none.fl_str_mv |
Montoya Estrada, María Paula Rincón González, Rincón González |
| dc.subject.proposal.spa.fl_str_mv |
Polimorfismos Deficiencia de Mieloperoxidasa Análisis Bioinformático PCR Secuenciación Candidiasis vulvovaginal Candida spp |
| topic |
Polimorfismos Deficiencia de Mieloperoxidasa Análisis Bioinformático PCR Secuenciación Candidiasis vulvovaginal Candida spp |
| description |
La mieloperoxidasa (MPO) es una proteína antimicrobiana presente en los gránulos azurófilos de los neutrófilos, su acción recae cuando se conjuga en el fagosoma con peróxido de hidrógeno, resultando en sustancias tóxicas para los microorganismos fagocitados (1). Se ha descrito que las infecciones vulvovaginales recurrentes por Candida spp tienen relación con la deficiencia priMaría de MPO, el cual es un trastorno autosómico recesivo (2), que presenta polimorfismos asociados al procesamiento postraduccional defectuoso de la proteína precursora o con errores pretraduccionales (3), por esto, el propósito fue identificar la relación que existe entre dos polimorfismos de la MPO (Y173C y M251T) con una mayor probabilidad de presentar este tipo de infecciones, puesto que este tipo de polimorfismos no han sido estudiados en el país. Frente a la metodología, se realizó una encuesta, donde se recolectaron 27 muestras de mujeres entre 20-45 años, se les realizó cuadro hemático, tinción Wright y tinción de peroxidasa leucocitaria. Se prosiguió con la extracción de ADN utilizando el kit Wizard® Genomic DNA Purification de Promega, luego se realizó la cuantificación del ADN extraído por espectrofotometría en NanoDrop, y la evaluación de la integridad del ADN mediante electroforesis. Por otro lado, se diseñaron los primers haciendo uso de herramientas bioinformáticas para la estandarización de la PCR convencional, posteriormente se usaron enzimas de restricción y se secuenciaron los productos amplificados con el fin de identificar los polimorfismos mencionados previamente. |
| publishDate |
2022 |
| dc.date.accessioned.none.fl_str_mv |
2022-10-18T17:27:13Z |
| dc.date.available.none.fl_str_mv |
2022-10-18T17:27:13Z |
| dc.date.issued.none.fl_str_mv |
2022-04-08 |
| 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/TP |
| 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/5717 |
| url |
https://repositorio.unicolmayor.edu.co/handle/unicolmayor/5717 |
| dc.language.iso.spa.fl_str_mv |
spa |
| language |
spa |
| dc.relation.references.spa.fl_str_mv |
NIH. Definición de inmunidad innata - Diccionario de cáncer del NCI - Instituto Nacional del Cáncer [Internet]. INMUNIDAD INNATA. [cited 2021 Aug 8]. Available from: https://www.cancer.gov/espanol/publicaciones/diccionarios/diccionariocancer/def/inmunidad-innata Aratani Y. Myeloperoxidase: Its role for host defense, inflammation, and neutrophil function. Arch Biochem Biophys. 2018;640(November 2017):47–52. Van Der Veen BS, De Winther MPJ, Heeringa P. Myeloperoxidase: Molecular mechanisms of action and their relevance to human health and disease. Antioxidants Redox Signal. 2009;11(11):2899–937. Nauseef WM. How human neutrophils kill and degrade microbes: An integrated view. Immunol Rev. 2007;219(1):88–102. Segal AW. How neutrophils kill microbes. Annu Rev Immunol. 2005;23(2):197–223. Kolaczkowska E, Kubes P. Neutrophil recruitment and function in health and inflammation. Nat Rev Immunol [Internet]. 2013;13(3):159–75. Available from: http://dx.doi.org/10.1038/nri3399 Staines Boone AT, González Serrano ME, Guidos Morales HE, Hernández Bautista VM, Espinosa Rosales FJ, Espinosa Padilla SE. Inmunodeficiencias por alteraciones en las células fagocitarias.pdf. Medigraphic. 2005;14. Henderson WR, Chi EY, Jong EC, Klebanoff SJ. Mast cell-mediated tumor-cell cytotoxicity. Role of the peroxidase system. J Exp Med. 1981;153(3):520–33. Khan AA, Rahmani AH, Aldebasi YH, Aly SM. Biochemical and pathological studies on peroxidases -an updated review. Glob J Health Sci. 2014;6(5):87–98 Nauseef WM. Biosynthesis of human myeloperoxidase. Arch Biochem Biophys [Internet]. 2018;642:1–9. Available from: https://doi.org/10.1016/j.abb.2018.02.001 Miyasaki KT, Nemirovskiy E. Myeloperoxidase isoform activities released by human neutrophils in response to dental and periodontal bacteria. Oral Microbiol Immunol. 1997;12(1):27–32 NCBI. MPO mieloperoxidasa [Homo sapiens (humano)] - Gen - NCBI [Internet]. 2014 [cited 2021 Aug 8]. Available from: https://www.ncbi.nlm.nih.gov/gene/4353#geneexpression DeLeo FR, Goedken M, McCormick SJ, Nauseef WM. A novel form of hereditary myeloperoxidase deficiency linked to endoplasmic reticulum/proteasome degradation. J Clin Invest. 1998;101(12):2900–9. Grishkovskaya XI, Paumann-Page M, Tscheliessnig R, Stampler J, Hofbauer XS, Soudi M, et al. Structure of human promyeloperoxidase (proMPO) and the role of the propeptide in processing and maturation. J Biol Chem. 2017;292(20):8244–61 Van Antwerpen P, Slomianny MC, Boudjeltia KZ, Delporte C, Faid V, Calay D, et al. Glycosylation pattern of mature dimeric leukocyte and recombinant monomeric myeloperoxidase: Glycosylation is required for optimal enzymatic activity. J Biol Chem. 2010;285(21):16351–9. Beckman Coulter. Mieloperoxidasa - Beckman Coulter [Internet]. [cited 2021 Aug 8]. Available from: https://www.mybeckman.co/reagents/coulter-flow-cytometry/antibodiesand-kits/single-color-antibodies/myeloperoxidase Khan A, Alsahli M, Rahmani A. Myeloperoxidase as an Active Disease Biomarker: Recent Biochemical and Pathological Perspectives. Med Sci. 2018;6(2):33 Kinkade JM. Evidence for the Involvement of an Acidic Compartment in the Processing of Myeloperoxidase in Human Promyelocytic Leukemia HL-60 Cells ’ ronment ( 10 ). Lysosomes are generally considered to be acidic organelles , and pro- teolysis involved in the product. 1987;255(2):428–36. Zhu J De. Myeloidcell-lineage and premylocutic-stage-specific- expression of the mouse myeloperoxidase gene is controlled at initiation as well as elongation levels of transcription. Cell Res. 1999;9(2):107–34. Nauseef WM, McCormick SJ, Goedken M. Coordinated participation of calreticulin and calnexin in the biosynthesis of myeloperoxidase. J Biol Chem. 1998;273(12):7107–11. Fiedler TJ, Davey CA, Fenna RE. X-ray crystal structure and characterization of halidebinding sites of human myeloperoxidase at 1.8 Å resolution. J Biol Chem. 2000;275(16):11964–71. García Morales OH, Roche NP, Flores Sánchez RM. Enzimas generadoras de especies reactivas del oxígeno: Mieloperoxidasa. Rev Cuba Investig Biomed. 1998;17(3):190–7 Reynolds WF, Rhees J, Maciejewski D, Paladino T, Sieburg H, Maki RA, et al. Myeloperoxidase polymorphism is associated with gender specific risk for Alzheimer’s disease. Exp Neurol. 1999;155(1):31–41. Crawford FC, Freeman MJ, Schinka JA, Morris MD, Abdullah LI, Richards D, et al. Association between Alzheimer’s disease and a functional polymorphism in the Myeloperoxidase gene. Exp Neurol. 2001;167(2):456–9 Willems HME, Ahmed SS, Liu J, Xu Z, Peters BM. Vulvovaginal candidiasis: A current understanding and burning questions. J Fungi. 2020;6(1). Homme M, Tateno N, Miura N, Ohno N, Aratani Y. Myeloperoxidase deficiency in mice exacerbates lung inflammation induced by nonviable Candida albicans. Inflamm Res. 2013;62(11):981–90. Persad AS, Kameoka Y, Kanda S, Niho Y, Suzuki K. Arginine to cysteine mutation (R499C) found in a japanese patient with complete myeloperoxidase deficiency. Gene Expr. 2005;13(2):67–71. Odobasic D, Kitching AR, Holdsworth SR. Neutrophil-mediated regulation of innate and adaptive immunity: The role of myeloperoxidase. J Immunol Res. 2016;2016. Kutter D, Devaquet P, Vanderstocken G, Paulus JM, Marchal V, Gothot A. Consequences of total and subtotal myeloperoxidase deficiency: Risk or benefit? Acta Haematol. 2000;104(1):10–5. Kutter D. Prevalence of myeloperoxidase deficiency: Population studies using BayerTechnicon automated hematology. J Mol Med. 1998;76(10):669–75. Zielinska-Jenczylik J, Sypula A, Budko E, Rzadkowska-Bodalska H. Interferonogenic and antiviral effect of extracts from Erodium cicutarium. Arch Immunol Ther Exp (Warsz). 1987;35(2):211–20 Pahwa R, Modi P JI. Myeloperoxidase Deficiency. tatPearls Publ. 2020; Nauseef WM, McCormick S, Goedken M. Impact of missense mutation on biosynthesis of myeloperoxidase. Redox Rep. 2000;5(4):197–206 Rutkowski R, A Pancewicz S, Rutkowski K, Rutkowska J. [Reactive oxygen and nitrogen species in inflammatory process]. Pol Merkur Lek [Internet]. 2007;134:131–6. Available from: https://pubmed.ncbi.nlm.nih.gov/18044345/ Freeman TA, Parvizi J, Della Valle CJ, Steinbeck MJ. Reactive oxygen and nitrogen species induce protein and DNA modifications driving arthrofibrosis following total knee arthroplasty. Fibrogenes Tissue Repair. 2009;2(1):1–11. Freeman TA, Parvizi J, Della Valle CJ, Steinbeck MJ. Reactive oxygen and nitrogen species induce protein and DNA modifications driving arthrofibrosis following total knee arthroplasty. Fibrogenes Tissue Repair. 2009;2(1):1–11. Freeman TA, Parvizi J, Della Valle CJ, Steinbeck MJ. Reactive oxygen and nitrogen species induce protein and DNA modifications driving arthrofibrosis following total knee arthroplasty. Fibrogenes Tissue Repair. 2009;2(1):1–11. Cos Padrón Y, Torres Leyva I, Marsán Suárez V, Macías Abraham C. Revista Cubana de Hematología, Inmunología y Hemoterapia. Rev Cuba Hematol Inmunol Hemoter [Internet]. 2004;20(0864–0289). Available from: http://scielo.sld.cu/scielo.php?script=sci_arttext&pid=S0864- 02892004000100002&lng=es&nrm=iso&tlng=es Kalinski T, Jentsch-Ullrich K, Fill S, König B, Costa SD, Roessner A. Lethal candida sepsis associated with myeloperoxidase deficiency and pre-eclampsia: Case report. Apmis. 2007;115(7):875–80. Netea MG, Maródi L. Innate immune mechanisms for recognition and uptake of Candida species. Trends Immunol. 2010;31(9):346–53. Rosati D, Bruno M, Jaeger M, Ten Oever J, Netea MG. Recurrent vulvovaginal candidiasis: An immunological perspective. Microorganisms. 2020;8(2):1–14. Sobel JD. Vulvovaginal candidosis. Lancet. 2007;369(9577):1961–71. Uppuluri P, Lopez Ribot JL. Candida albicans biofilms. Candida albicans Cell Mol Biol Second Ed. 2017;18(5):63–75. Uppuluri P, Lopez Ribot JL. Candida albicans biofilms. Candida albicans Cell Mol Biol Second Ed. 2017;18(5):63–75. Mbakwem-Aniebo C, Osadebe AU, Athanasonny E, Okonko IO. Prevalence of Candida spp. and age-related disparities amongst women presenting with vaginitis at the obstetrics and gynaecology (O&G) clinic in a tertiary hospital in Port Harcourt, Nigeria. Afr Health Sci. 2020;20(1):51–8. Beltrame A, Matteelli A, Carvalho ACC, Saleri N, Casalini C, Capone S, et al. Vaginal colonization with Candida spp. in human immunodeficiency virus - Infected women: A cohort study. Int J STD AIDS. 2006;17(4):260–6 Vermitsky JP, Self MJ, Chadwick SG, Trama JP, Adelson ME, Mordechai E, et al. Survey of vaginal-flora Candida species isolates from women of different age groups by use of species-specific PCR detection. J Clin Microbiol. 2008;46(4):1501–3. Bradford LL, Ravel J. The vaginal mycobiome: A contemporary perspective on fungi in women’s health and diseases. Virulence. 2017;8(3):342–51. Bitew A, Abebaw Y. Vulvovaginal candidiasis: Species distribution of Candida and their antifungal susceptibility pattern. BMC Womens Health. 2018;18(1):1–10. Aratani Y, Kura F, Watanabe H, Akagawa H, Takano Y, Suzuki K, et al. Critical role of myeloperoxidase and nicotinamide adenine dinucleotide phosphate-oxidase in high-burden systemic infection of mice with Candida albicans. J Infect Dis. 2002;185(12):1833–7 Aratani Y, Kura F, Watanabe H, Akagawa H, Takano Y, Suzuki K, et al. Relative contributions of myeloperoxidase and NADPH-oxidase to the early host defense against 45 pulmonary infections with Candida albicans and Aspergillus fumigatus. Med Mycol. 2002;40(6):557–63 Lina Marcela RP. Identificación de polimorfismo(s) en el gen de la Mieloperoxidasa asociado a infecciones recurrentes por . Universidad Colegio Mayor de Cundinamarca; 2018 Clark BRA, Klebanoff SJ, System PC. Brief Definitive Report NEUTROPHILMEDIATED TUMOR CELL CYTOTOXICITY : ROLE OF THE PEROXIDASE SYSTEM * Polymorphonuclear leukocytes ( PMN ) employ a system comprised of myeloperoxidase ( MPO ), H20 2 , and an oxidizable halide cofactor to kill a variety . 1975;141:1442–7 Montpellier TS. Br. J. exp. Path (1982) 63, 320 SERUM INHIBITORY EFFECT ON MYELOPEROXIDASE IN HUMAN CANCER. 1982;320–4. Antachopoulos C. 2010 (Antachopoulos) Invasive fungal infections in congenital immunodeficiencies.pdf. 2010;1. Plantinga TS, Johnson MD, Scott WK, Van De Vosse E, Velez Edwards DR, Smith PB, et al. Toll-like receptor 1 polymorphisms increase susceptibility to candidemia. J Infect Dis. 2012;205(6):934–43 Cassone A. Vulvovaginal Candida albicans infections: Pathogenesis, immunity and vaccine prospects. BJOG An Int J Obstet Gynaecol. 2015;122(6):785–94 Vélez G, Rocha Y, Arias A, López J. Función del sistema NADPH oxidasa en la formación de trampas extracelulares de los neutrófilos (NETs). Rev Cuba Hematol Inmunol y Hemoter. 2016;32(1):43–56. Nauseef WM. Myeloperoxidase in human neutrophil host defence. Cell Microbiol. 2014;16(8):1146–55. Beckman Coulter. Mieloperoxidasa - Beckman Coulter [Internet]. Antigeno de mieloperoxidasa. [cited 2021 Aug 23]. Available from: https://www.mybeckman.co/reagents/coulter-flow-cytometry/antibodies-and-kits/singlecolor-antibodies/myeloperoxidase Nagra RM, Becher B, Tourtellotte WW, Antel JP, Gold D, Paladino T, et al. Immunohistochemical and genetic evidence of myeloperoxidase involvement in multiple sclerosis. J Neuroimmunol. 1997;78(1–2):97–107. Hoy A, Leininger-Muller B, Kutter D, Siest G, Visvikis S. Growing significance of myeloperoxidase in non-infectious diseases. Clin Chem Lab Med. 2002;40(1):2–8 Green PS, Mendez AJ, Jacob JS, Crowley JR, Growdon W, Hyman BT, et al. Neuronal expression of myeloperoxidase is increased in Alzheimer’s disease. J Neurochem. 2004;90(3):724–33. Nauseef WM, Cogley M, McCormick S. Effect of the R569W missense mutation on the biosynthesis of myeloperoxidase. J Biol Chem. 1996;271(16):9546–9 Ohashi YY, Kameoka Y, Persad AS, Koi F, Yamagoe S, Hashimoto K, et al. Novel missense mutation found in a Japanese patient with myeloperoxidase deficiency. Gene. 2004;327(2):195–200. Aratani Y, Koyama H, Nyui SI, Suzuki K, Kura F, Maeda N. Severe impairment in early host defense against Candida albicans in mice deficient in myeloperoxidase. Infect Immun. 1999;67(4):1828–36. Marchetti C, Patriarca P, Solero G Pietro, Baralle FE, Romano M. Genetic Characterization of Myeloperoxidase Deficiency in Italy. Hum Mutat. 2004;23(5):496–505 Comino R, Davi E, Martı JC, Docente U, Gene D De. OBSTETRICIA Y GINECOLOG ´ La vulvovaginitis candidia. 2013;56(2):108–16. Available from: file:///C:/Users/Usuario/Downloads/S0304501312001641_S300_es.pdf Pemán J, Quindós G. Aspectos actuales de las enfermedades invasoras causadas por Candida y otros hongos levaduriformes. Rev Iberoam Micol [Internet]. 2016;33(3):133–9. Available from: http://dx.doi.org/10.1016/j.riam.2015.10.001 Pemán J, Quindós G. Aspectos actuales de las enfermedades invasoras causadas por Candida y otros hongos levaduriformes. Rev Iberoam Micol [Internet]. 2016;33(3):133–9. Available from: http://dx.doi.org/10.1016/j.riam.2015.10.001 Kwak W, Han YH, Seol D, Kim H, Ahn H, Jeong M, et al. Complete Genome of Lactobacillus iners KY Using Flongle Provides Insight Into the Genetic Background of Optimal Adaption to Vaginal Econiche. Front Microbiol. 2020;11(May):1–10. Aldunate M, Srbinovski D, Hearps AC, Latham CF, Ramsland PA, Gugasyan R, et al. Antimicrobial and immune modulatory effects of lactic acid and short chain fatty acids produced by vaginal microbiota associated with eubiosis and bacterial vaginosis. Front Physiol. 2015;6(JUN):1–23 Parolin C, Marangoni A, Laghi L, Foschi C, Palomino RAÑ, Calonghi N, et al. Isolation of vaginal lactobacilli and characterization of anti-candida activity. PLoS One. 2015;10(6):1– 17. Van De Wijgert JHHM, Borgdorff H, Verhelst R, Crucitti T, Francis S, Verstraelen H, et al. The vaginal microbiota: What have we learned after a decade of molecular characterization? PLoS One. 2014;9(8). Li J, McCormick J, Bocking A, Reid G. Importance of vaginal microbes in reproductive health. Reprod Sci. 2012;19(3):235–42. Li J, McCormick J, Bocking A, Reid G. Importance of vaginal microbes in reproductive health. Reprod Sci. 2012;19(3):235–42. Sha BE, Zariffard MR, Wang QJ, Chen HY, Bremer J, Cohen MH, et al. Female genitaltract HIV load correlates inversely with Lactobacillus species but positively with bacterial vaginosis and Mycoplasma hominis. J Infect Dis. 2005;191(1):25–32. Hayes R, Watson-Jones D, Celum C, Van De Wijgert J, Wasserheit J. Treatment of sexually transmitted infections for HIV prevention: End of the road or new beginning? Aids. 2010;24(SUPPL. 4):1–19 Van De Wijgert JHHM, Morrison CS, Cornelisse PGA, Munjoma M, Moncada J, Awio P, et al. Bacterial vaginosis and vaginal yeast, but not vaginal cleansing, increase HIV-1 acquisition in African women. J Acquir Immune Defic Syndr. 2008;48(2):203–10. Jacobsen ID, Hube B. Candida albicans morphology: still in focus. Expert Rev Anti Infect Ther [Internet]. 2017;15(4):327–30. Available from: http://dx.doi.org/10.1080/14787210.2017.1290524 Poulain D. Candida albicans, plasticity and pathogenesis. Crit Rev Microbiol. 2015;41(2):208–17. Szafranski-Schneider E, Swidergall M, Cottier F, Tielker D, Román E, Pla J, et al. Msb2 shedding protects Candida albicans against antimicrobial peptides. PLoS Pathog. 2012;8(2) Spettel K, Barousch W, Makristathis A, Zeller I, Nehr M, Selitsch B, et al. Analysis of antifungal resistance genes in Candida albicans and Candida glabrata using next generation sequencing. PLoS One. 2019;14(1):1–19. Gao J, Wang H, Li Z, Wong AHH, Wang YZ, Guo Y, et al. Candida albicans gains azole resistance by altering sphingolipid composition. Nat Commun [Internet]. 2018;9(1):1–14. Available from: http://dx.doi.org/10.1038/s41467-018-06944-1 Silva S, Negri M, Henriques M, Oliveira R, Williams DW, Azeredo J. Candida glabrata, Candida parapsilosis and Candida tropicalis: Biology, epidemiology, pathogenicity and antifungal resistance. FEMS Microbiol Rev. 2012;36(2):288–305 Ramos JT, Francisco L, Daoud Z. Infección fúngica invasora en niños: diferencias y analogías con el adulto. Rev Española Quimioter. 2016;29(1):59–65. Wu EY, Ehrlich L, Handly B, Frush DP, Rebecca H, Hill C, et al. Immunodeficiency Disorders : an Update. 2017;46(12):1630–44. Quindós G. Epidemiology of candidaemia and invasive candidiasis. A changing face. Rev Iberoam Micol [Internet]. 2014;31(1):42–8. Available from: http://dx.doi.org/10.1016/j.riam.2013.10.001 Sanches JM, Giraldo PC, Bardin MG, Amaral R, Discacciati MG, Rossato L. Laboratorial Aspects of Cytolytic Vaginosis and Vulvovaginal Candidiasis as a Key for Accurate Diagnosis: A Pilot Study. Rev Bras Ginecol e Obstet. 2020;42(10):634–41. Uribarren-Berrueta T del NJ, Pineda-Murillo J, Cortés-Figueroa AÁ, Castañon Olivares LR. Candidosis vaginal. Segunda parte:diagnóstico, tratamiento y situación de algunos países latinoamericanos. Rev Médica Risaralda. 2017;23(1):159–66. Farr A, Effendy I, Frey Tirri B, Hof H, Mayser P, Petricevic L, et al. Guideline: Vulvovaginal candidosis (AWMF 015/072, level S2k). Mycoses. 2021;64(6):583–602. Pereira LC, Correia AF, da Silva ZDL, de Resende CN, Brandão F, Almeida RM, et al. Vulvovaginal candidiasis and current perspectives: new risk factors and laboratory diagnosis by using MALDI TOF for identifying species in primary infection and recurrence. Eur J Clin Microbiol Infect Dis. 2021;40(8):1681–93 Candidosis vaginal - Diagnóstico y tratamiento - Mayo Clinic [Internet]. [cited 2021 Sep 5]. Available from: https://www.mayoclinic.org/es-es/diseases-conditions/yeastinfection/diagnosis-treatment/drc-20379004 Colomer M. Candidiasis vulvovaginal Tratamiento tópico y oral. Actual científica Av Farm. 2009;28(1):68–9 Pérez AM. Reacción en cadena de la polimerasa (Polymerase Chain Reaction, PCR). Repos Inst la Univ Politècnica València [Internet]. 2011;10. Available from: https://riunet.upv.es/bitstream/handle/10251/10700/Reacción en cadena de la polimerasa.pdf Tamay de Dios L * Ibarra C,** Velasquillo C*. Fundamentos de la reacción en cadena de la polimerasa (PCR) y de la PCR en tiempo real. Medigraphic [Internet]. 2013 [cited 2021 Sep 5];2:70–8. Available from: www.medigraphic.org.mxhttp://www.medigraphic.com/ridwww.medigraphic.org.mx Márquez L, Serrato A, Cerritos R. Secuenciación de fragmentos de ADN. Herramientas Mol Apl en Ecol Asp teóricos y prácticos [Internet]. 2014;231–49. Available from: http://www2.inecc.gob.mx/publicaciones/libros/710/secuenciacion.pdf Carla LC, Fernando GC, María TC, Antonio OP, Emilia CM, Rafael CM. Aplicaciones de las técnicas de secuenciación masiva en la Microbiología Clínica [Internet]. 2021. 1–55 p. Available from: https://seimc.org/contenidos/documentoscientificos/procedimientosmicrobiologia/seimc-procedimiento71.pdf Products A. Manual de funcionamiento BC 3000 plus Hematologia. Maquin Mont LTDA [Internet]. 2014;(7001):1–43. Available from: http://www.maquinariamontana.com/images/manuales/H616 618 620.pdf%0Ahttp://sistemamid.com/preview.php?a=6209 NCBI. Cromosoma 17 de Homo sapiens, GRCh38.p13 Ensamblaje primario - Nucleótido - NCBI [Internet]. [cited 2021 Sep 6]. Available from: https://www.ncbi.nlm.nih.gov/nuccore/NC_000017.11?report=genbank&from=58269855& to=58280935&strand=true Campos-Sanchez E, Martínez-Cano J, del Pino Molina L, López-Granados E, Cobaleda C. Epigenetic Deregulation in Human Primary Immunodeficiencies. Trends Immunol. 2019;40(1):49–65 Capucine P, Bousfiha A, Jeddane L, Ailal F, Gaspar HB, Crow WA-H, et al. The 2017 IUIS Phenotypic Classification for Primary Immunodeficiencies. Lubr Eng. 1997;53(7):12–7 Lanza F. Clinical manifestation of myeloperoxidase deficiency. J Mol Med. 1998;76(10):676–81. Carmel R, Watkins D, Rosenblatt DS. Chapter 10 – Megaloblastic Anemia. Nathan Oski’s Hematol Oncol Infancy Child. 8th Ed. 2015;308-343.e11. Cheng G, Salerno JC, Cao Z, Pagano PJ, Lambeth JD. Identification and characterization of VPO1, a new animal heme-containing Peroxidase. El-Ashram S, Al Nasr I, Suo X. Nucleic acid protocols: Extraction and optimization. Biotechnol Reports [Internet]. 2016;12:33–9. Available from: http://dx.doi.org/10.1016/j.btre.2016.10.001 Metzler KD, Fuchs TA, Nauseef WM, Reumaux D, Roesler J, Schulze I, et al. Myeloperoxidase is required for neutrophil extracellular trap formation: Implications for innate immunity. Blood. 2011;117(3):953–9 Quie PG, White JG, Holmes B, Good RA. In vitro bactericidal capacity of human polymorphonuclear leukocytes: diminished activity in chronic granulomatous disease of childhood. J Clin Invest. 1967;46(4):668–79 Koziol-Montewka M, Magrys A, Paluch-Oles J, Bogut A, Buczynski K, Jablonka S. MPO and cytokines in the serum of cancer patients in the context of Candida colonization and infection. Immunol Invest. 2006;35(2):167–79. Vigezzi C, Riera FO, Rodriguez E, Icely PA, Miró MS, Figueredo CM, et al. Candidiasis invasora: un enfoque a la infección en el sistema nervioso central. Rev Argent Microbiol. 2021;53(2):171–8 Serrato Díaz A, Flores Rentería L, Aportela Cortez J, Sierra Palacios E. PCR: reacción en cadena de la polimerasa. [cited 2021 Sep 5]; Available from: www.biologia |
| 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 |
83p. |
| 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/5717/4/license.txt https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5717/5/Identificaci%c3%b3n%20de%20polimorfismos%20en%20el%20gen%20mieloperoxidasa%20y%20su%20papel%20en%20candidiasis%20vulvovaginal%20recurrente%20por%20Candida%20spp%20en%20pacientes%20de%20Bogot%c3%a1%20DC.pdf.txt https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5717/7/4.%20FORMATO%20IDENTIFICACI%c3%93N%20TRABAJOS%20DE%20GRADO%281%29.pdf.txt https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5717/9/CARTA%20DERECHOS%20DE%20AUTOR%20Montoya.pdf.txt https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5717/6/Identificaci%c3%b3n%20de%20polimorfismos%20en%20el%20gen%20mieloperoxidasa%20y%20su%20papel%20en%20candidiasis%20vulvovaginal%20recurrente%20por%20Candida%20spp%20en%20pacientes%20de%20Bogot%c3%a1%20DC.pdf.jpg https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5717/8/4.%20FORMATO%20IDENTIFICACI%c3%93N%20TRABAJOS%20DE%20GRADO%281%29.pdf.jpg https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5717/10/CARTA%20DERECHOS%20DE%20AUTOR%20Montoya.pdf.jpg https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5717/11/Identificaci%c3%b3n%20de%20polimorfismos%20en%20el%20gen%20mieloperoxidasa%20y%20su%20papel%20en%20candidiasis%20vulvovaginal%20recurrente%20por%20Candida%20spp%20en%20pacientes%20de%20Bogot%c3%a1%20DC.pdf https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5717/12/CARTA%20DERECHOS%20DE%20AUTOR%20Montoya.pdf https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5717/13/4.%20FORMATO%20IDENTIFICACI%c3%93N%20TRABAJOS%20DE%20GRADO%281%29.pdf |
| bitstream.checksum.fl_str_mv |
2f9959eaf5b71fae44bbf9ec84150c7a 801d2b9a1135c7975a16de8056561107 e05e93a1452a4c335cdf98fd5236eb38 123eb2349f8a77bfa64c6bdf27fd125c a7bea7b6fae6c8aec9b80437c67fb999 ed029c2618f978e938d4434cdebd7e9e 646cd9b36ffbf7f3c752978f84bd4224 437853c9633a805b5cb1520eb9b14303 4b0e49e2b81b201c7954c7d506fbee93 aa95c18a0f56d7461d118693f309aebc |
| bitstream.checksumAlgorithm.fl_str_mv |
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_ |
1812210091180425216 |
| spelling |
Cruz Baquero, Claudia Andreaf37a550d32dcc1622c0035ee0120dd76Pinilla Bermúdez, Gladys428561c7b44bd4a7c33f25d8f79d473c600Montoya Estrada, María Paulaf87f1d2683ab230ba68838d4201f05eeRincón González, Rincón González97bcf0361360f411d6bff0baa861b37f2022-10-18T17:27:13Z2022-10-18T17:27:13Z2022-04-08https://repositorio.unicolmayor.edu.co/handle/unicolmayor/5717La mieloperoxidasa (MPO) es una proteína antimicrobiana presente en los gránulos azurófilos de los neutrófilos, su acción recae cuando se conjuga en el fagosoma con peróxido de hidrógeno, resultando en sustancias tóxicas para los microorganismos fagocitados (1). Se ha descrito que las infecciones vulvovaginales recurrentes por Candida spp tienen relación con la deficiencia priMaría de MPO, el cual es un trastorno autosómico recesivo (2), que presenta polimorfismos asociados al procesamiento postraduccional defectuoso de la proteína precursora o con errores pretraduccionales (3), por esto, el propósito fue identificar la relación que existe entre dos polimorfismos de la MPO (Y173C y M251T) con una mayor probabilidad de presentar este tipo de infecciones, puesto que este tipo de polimorfismos no han sido estudiados en el país. Frente a la metodología, se realizó una encuesta, donde se recolectaron 27 muestras de mujeres entre 20-45 años, se les realizó cuadro hemático, tinción Wright y tinción de peroxidasa leucocitaria. Se prosiguió con la extracción de ADN utilizando el kit Wizard® Genomic DNA Purification de Promega, luego se realizó la cuantificación del ADN extraído por espectrofotometría en NanoDrop, y la evaluación de la integridad del ADN mediante electroforesis. Por otro lado, se diseñaron los primers haciendo uso de herramientas bioinformáticas para la estandarización de la PCR convencional, posteriormente se usaron enzimas de restricción y se secuenciaron los productos amplificados con el fin de identificar los polimorfismos mencionados previamente.TABLA DE CONTENIDO RESUMEN. 12 INTRODUCCIÓN . 13 1. ANTECEDENTES DEL PROYECTO .. 17 2. MARCO REFERENCIAL... 18 2.1 Sistema inmune . 18 2.2 Neutrófilos.. 19 2.3 Mieloperoxidasa 20 2.4 Polimorfismos relacionados con la deficiencia de MPO. 21 2.5 Inmunodeficiencias... 22 2.6 Candida spp 22 2.7 Patogenia de Candida spp... 23 2.8 Generalidades de candidiasis.. 23 2.9 Epidemiología 24 2.10 Diagnóstico.. 24 2.11 Tratamiento 25 2.12 Deficiencia de Mieloperoxidasa y susceptibilidad a infecciones por Candida spp. 25 2.13 Diagnóstico deficiencia de MPO. 26 2.13.1 Técnicas citoquímicas.. 26 2.13.2 PCR convencional .. 27 2.13.3 Secuenciación de ADN. 27 3. DISEÑO METODOLÓGICO. 27 3.1 Universo, Población y Muestra... 27 3.2 Hipótesis y Variables... 28 3.3. Técnicas y procedimientos 28 3.3.1 Recolección de muestras de sangre 28 3.3.2 Consentimiento informado .. 28 3.3.3 Análisis parámetros hemáticos ... 28 3.3.4 Identificación de peroxidasa leucocitaria 29 3.3.5 Identificación del gen mpo 29 3.3.6 Enzimas de restricción y electroforesis en gel de agarosa .. 30 3.3.7 Detección de polimorfismo del gen mpo... 30 4. RESULTADOS 31 4.1 Análisis de parámetros hemáticos 31 4.2 Identificación del gen mpo. 31 4.3 Enzimas de restricción y electroforesis en gel de agarosa. 33 4.4 Detección de polimorfismos del gen mpo.. 33 4 DISCUSIÓN. 34 5 CONCLUSIONES.. 39 REFERENCIAS... 41PregradoBacteriólogo(a) y Laboratorista Clínico83p.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_14cbIdentificación de polimorfismos en el gen Mieloperoxidasa y su papel en Candidiasis Vulvovaginal recurrente por Candida SPP en pacientes de Bogotá DCTrabajo 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/TPinfo:eu-repo/semantics/publishedVersionFacultad de Ciencias de la SaludBogotáBacteriología y Laboratorio ClínicoNIH. Definición de inmunidad innata - Diccionario de cáncer del NCI - Instituto Nacional del Cáncer [Internet]. INMUNIDAD INNATA. [cited 2021 Aug 8]. Available from: https://www.cancer.gov/espanol/publicaciones/diccionarios/diccionariocancer/def/inmunidad-innataAratani Y. Myeloperoxidase: Its role for host defense, inflammation, and neutrophil function. Arch Biochem Biophys. 2018;640(November 2017):47–52.Van Der Veen BS, De Winther MPJ, Heeringa P. Myeloperoxidase: Molecular mechanisms of action and their relevance to human health and disease. Antioxidants Redox Signal. 2009;11(11):2899–937.Nauseef WM. How human neutrophils kill and degrade microbes: An integrated view. Immunol Rev. 2007;219(1):88–102.Segal AW. How neutrophils kill microbes. Annu Rev Immunol. 2005;23(2):197–223.Kolaczkowska E, Kubes P. Neutrophil recruitment and function in health and inflammation. Nat Rev Immunol [Internet]. 2013;13(3):159–75. Available from: http://dx.doi.org/10.1038/nri3399Staines Boone AT, González Serrano ME, Guidos Morales HE, Hernández Bautista VM, Espinosa Rosales FJ, Espinosa Padilla SE. Inmunodeficiencias por alteraciones en las células fagocitarias.pdf. Medigraphic. 2005;14.Henderson WR, Chi EY, Jong EC, Klebanoff SJ. Mast cell-mediated tumor-cell cytotoxicity. Role of the peroxidase system. J Exp Med. 1981;153(3):520–33.Khan AA, Rahmani AH, Aldebasi YH, Aly SM. Biochemical and pathological studies on peroxidases -an updated review. Glob J Health Sci. 2014;6(5):87–98Nauseef WM. Biosynthesis of human myeloperoxidase. Arch Biochem Biophys [Internet]. 2018;642:1–9. Available from: https://doi.org/10.1016/j.abb.2018.02.001Miyasaki KT, Nemirovskiy E. Myeloperoxidase isoform activities released by human neutrophils in response to dental and periodontal bacteria. Oral Microbiol Immunol. 1997;12(1):27–32NCBI. MPO mieloperoxidasa [Homo sapiens (humano)] - Gen - NCBI [Internet]. 2014 [cited 2021 Aug 8]. Available from: https://www.ncbi.nlm.nih.gov/gene/4353#geneexpressionDeLeo FR, Goedken M, McCormick SJ, Nauseef WM. A novel form of hereditary myeloperoxidase deficiency linked to endoplasmic reticulum/proteasome degradation. J Clin Invest. 1998;101(12):2900–9.Grishkovskaya XI, Paumann-Page M, Tscheliessnig R, Stampler J, Hofbauer XS, Soudi M, et al. Structure of human promyeloperoxidase (proMPO) and the role of the propeptide in processing and maturation. J Biol Chem. 2017;292(20):8244–61Van Antwerpen P, Slomianny MC, Boudjeltia KZ, Delporte C, Faid V, Calay D, et al. Glycosylation pattern of mature dimeric leukocyte and recombinant monomeric myeloperoxidase: Glycosylation is required for optimal enzymatic activity. J Biol Chem. 2010;285(21):16351–9.Beckman Coulter. Mieloperoxidasa - Beckman Coulter [Internet]. [cited 2021 Aug 8]. Available from: https://www.mybeckman.co/reagents/coulter-flow-cytometry/antibodiesand-kits/single-color-antibodies/myeloperoxidaseKhan A, Alsahli M, Rahmani A. Myeloperoxidase as an Active Disease Biomarker: Recent Biochemical and Pathological Perspectives. Med Sci. 2018;6(2):33Kinkade JM. Evidence for the Involvement of an Acidic Compartment in the Processing of Myeloperoxidase in Human Promyelocytic Leukemia HL-60 Cells ’ ronment ( 10 ). Lysosomes are generally considered to be acidic organelles , and pro- teolysis involved in the product. 1987;255(2):428–36.Zhu J De. Myeloidcell-lineage and premylocutic-stage-specific- expression of the mouse myeloperoxidase gene is controlled at initiation as well as elongation levels of transcription. Cell Res. 1999;9(2):107–34.Nauseef WM, McCormick SJ, Goedken M. Coordinated participation of calreticulin and calnexin in the biosynthesis of myeloperoxidase. J Biol Chem. 1998;273(12):7107–11.Fiedler TJ, Davey CA, Fenna RE. X-ray crystal structure and characterization of halidebinding sites of human myeloperoxidase at 1.8 Å resolution. J Biol Chem. 2000;275(16):11964–71.García Morales OH, Roche NP, Flores Sánchez RM. Enzimas generadoras de especies reactivas del oxígeno: Mieloperoxidasa. Rev Cuba Investig Biomed. 1998;17(3):190–7Reynolds WF, Rhees J, Maciejewski D, Paladino T, Sieburg H, Maki RA, et al. Myeloperoxidase polymorphism is associated with gender specific risk for Alzheimer’s disease. Exp Neurol. 1999;155(1):31–41.Crawford FC, Freeman MJ, Schinka JA, Morris MD, Abdullah LI, Richards D, et al. Association between Alzheimer’s disease and a functional polymorphism in the Myeloperoxidase gene. Exp Neurol. 2001;167(2):456–9Willems HME, Ahmed SS, Liu J, Xu Z, Peters BM. Vulvovaginal candidiasis: A current understanding and burning questions. J Fungi. 2020;6(1).Homme M, Tateno N, Miura N, Ohno N, Aratani Y. Myeloperoxidase deficiency in mice exacerbates lung inflammation induced by nonviable Candida albicans. Inflamm Res. 2013;62(11):981–90.Persad AS, Kameoka Y, Kanda S, Niho Y, Suzuki K. Arginine to cysteine mutation (R499C) found in a japanese patient with complete myeloperoxidase deficiency. Gene Expr. 2005;13(2):67–71.Odobasic D, Kitching AR, Holdsworth SR. Neutrophil-mediated regulation of innate and adaptive immunity: The role of myeloperoxidase. J Immunol Res. 2016;2016.Kutter D, Devaquet P, Vanderstocken G, Paulus JM, Marchal V, Gothot A. Consequences of total and subtotal myeloperoxidase deficiency: Risk or benefit? Acta Haematol. 2000;104(1):10–5.Kutter D. Prevalence of myeloperoxidase deficiency: Population studies using BayerTechnicon automated hematology. J Mol Med. 1998;76(10):669–75.Zielinska-Jenczylik J, Sypula A, Budko E, Rzadkowska-Bodalska H. Interferonogenic and antiviral effect of extracts from Erodium cicutarium. Arch Immunol Ther Exp (Warsz). 1987;35(2):211–20Pahwa R, Modi P JI. Myeloperoxidase Deficiency. tatPearls Publ. 2020;Nauseef WM, McCormick S, Goedken M. Impact of missense mutation on biosynthesis of myeloperoxidase. Redox Rep. 2000;5(4):197–206Rutkowski R, A Pancewicz S, Rutkowski K, Rutkowska J. [Reactive oxygen and nitrogen species in inflammatory process]. Pol Merkur Lek [Internet]. 2007;134:131–6. Available from: https://pubmed.ncbi.nlm.nih.gov/18044345/Freeman TA, Parvizi J, Della Valle CJ, Steinbeck MJ. Reactive oxygen and nitrogen species induce protein and DNA modifications driving arthrofibrosis following total knee arthroplasty. Fibrogenes Tissue Repair. 2009;2(1):1–11.Freeman TA, Parvizi J, Della Valle CJ, Steinbeck MJ. Reactive oxygen and nitrogen species induce protein and DNA modifications driving arthrofibrosis following total knee arthroplasty. Fibrogenes Tissue Repair. 2009;2(1):1–11.Freeman TA, Parvizi J, Della Valle CJ, Steinbeck MJ. Reactive oxygen and nitrogen species induce protein and DNA modifications driving arthrofibrosis following total knee arthroplasty. Fibrogenes Tissue Repair. 2009;2(1):1–11.Cos Padrón Y, Torres Leyva I, Marsán Suárez V, Macías Abraham C. Revista Cubana de Hematología, Inmunología y Hemoterapia. Rev Cuba Hematol Inmunol Hemoter [Internet]. 2004;20(0864–0289). Available from: http://scielo.sld.cu/scielo.php?script=sci_arttext&pid=S0864- 02892004000100002&lng=es&nrm=iso&tlng=esKalinski T, Jentsch-Ullrich K, Fill S, König B, Costa SD, Roessner A. Lethal candida sepsis associated with myeloperoxidase deficiency and pre-eclampsia: Case report. Apmis. 2007;115(7):875–80.Netea MG, Maródi L. Innate immune mechanisms for recognition and uptake of Candida species. Trends Immunol. 2010;31(9):346–53.Rosati D, Bruno M, Jaeger M, Ten Oever J, Netea MG. Recurrent vulvovaginal candidiasis: An immunological perspective. Microorganisms. 2020;8(2):1–14.Sobel JD. Vulvovaginal candidosis. Lancet. 2007;369(9577):1961–71.Uppuluri P, Lopez Ribot JL. Candida albicans biofilms. Candida albicans Cell Mol Biol Second Ed. 2017;18(5):63–75.Uppuluri P, Lopez Ribot JL. Candida albicans biofilms. Candida albicans Cell Mol Biol Second Ed. 2017;18(5):63–75.Mbakwem-Aniebo C, Osadebe AU, Athanasonny E, Okonko IO. Prevalence of Candida spp. and age-related disparities amongst women presenting with vaginitis at the obstetrics and gynaecology (O&G) clinic in a tertiary hospital in Port Harcourt, Nigeria. Afr Health Sci. 2020;20(1):51–8.Beltrame A, Matteelli A, Carvalho ACC, Saleri N, Casalini C, Capone S, et al. Vaginal colonization with Candida spp. in human immunodeficiency virus - Infected women: A cohort study. Int J STD AIDS. 2006;17(4):260–6Vermitsky JP, Self MJ, Chadwick SG, Trama JP, Adelson ME, Mordechai E, et al. Survey of vaginal-flora Candida species isolates from women of different age groups by use of species-specific PCR detection. J Clin Microbiol. 2008;46(4):1501–3.Bradford LL, Ravel J. The vaginal mycobiome: A contemporary perspective on fungi in women’s health and diseases. Virulence. 2017;8(3):342–51.Bitew A, Abebaw Y. Vulvovaginal candidiasis: Species distribution of Candida and their antifungal susceptibility pattern. BMC Womens Health. 2018;18(1):1–10.Aratani Y, Kura F, Watanabe H, Akagawa H, Takano Y, Suzuki K, et al. Critical role of myeloperoxidase and nicotinamide adenine dinucleotide phosphate-oxidase in high-burden systemic infection of mice with Candida albicans. J Infect Dis. 2002;185(12):1833–7Aratani Y, Kura F, Watanabe H, Akagawa H, Takano Y, Suzuki K, et al. Relative contributions of myeloperoxidase and NADPH-oxidase to the early host defense against 45 pulmonary infections with Candida albicans and Aspergillus fumigatus. Med Mycol. 2002;40(6):557–63Lina Marcela RP. Identificación de polimorfismo(s) en el gen de la Mieloperoxidasa asociado a infecciones recurrentes por . Universidad Colegio Mayor de Cundinamarca; 2018Clark BRA, Klebanoff SJ, System PC. Brief Definitive Report NEUTROPHILMEDIATED TUMOR CELL CYTOTOXICITY : ROLE OF THE PEROXIDASE SYSTEM * Polymorphonuclear leukocytes ( PMN ) employ a system comprised of myeloperoxidase ( MPO ), H20 2 , and an oxidizable halide cofactor to kill a variety . 1975;141:1442–7Montpellier TS. Br. J. exp. Path (1982) 63, 320 SERUM INHIBITORY EFFECT ON MYELOPEROXIDASE IN HUMAN CANCER. 1982;320–4.Antachopoulos C. 2010 (Antachopoulos) Invasive fungal infections in congenital immunodeficiencies.pdf. 2010;1.Plantinga TS, Johnson MD, Scott WK, Van De Vosse E, Velez Edwards DR, Smith PB, et al. Toll-like receptor 1 polymorphisms increase susceptibility to candidemia. J Infect Dis. 2012;205(6):934–43Cassone A. Vulvovaginal Candida albicans infections: Pathogenesis, immunity and vaccine prospects. BJOG An Int J Obstet Gynaecol. 2015;122(6):785–94Vélez G, Rocha Y, Arias A, López J. Función del sistema NADPH oxidasa en la formación de trampas extracelulares de los neutrófilos (NETs). Rev Cuba Hematol Inmunol y Hemoter. 2016;32(1):43–56.Nauseef WM. Myeloperoxidase in human neutrophil host defence. Cell Microbiol. 2014;16(8):1146–55.Beckman Coulter. Mieloperoxidasa - Beckman Coulter [Internet]. Antigeno de mieloperoxidasa. [cited 2021 Aug 23]. Available from: https://www.mybeckman.co/reagents/coulter-flow-cytometry/antibodies-and-kits/singlecolor-antibodies/myeloperoxidaseNagra RM, Becher B, Tourtellotte WW, Antel JP, Gold D, Paladino T, et al. Immunohistochemical and genetic evidence of myeloperoxidase involvement in multiple sclerosis. J Neuroimmunol. 1997;78(1–2):97–107.Hoy A, Leininger-Muller B, Kutter D, Siest G, Visvikis S. Growing significance of myeloperoxidase in non-infectious diseases. Clin Chem Lab Med. 2002;40(1):2–8Green PS, Mendez AJ, Jacob JS, Crowley JR, Growdon W, Hyman BT, et al. Neuronal expression of myeloperoxidase is increased in Alzheimer’s disease. J Neurochem. 2004;90(3):724–33.Nauseef WM, Cogley M, McCormick S. Effect of the R569W missense mutation on the biosynthesis of myeloperoxidase. J Biol Chem. 1996;271(16):9546–9Ohashi YY, Kameoka Y, Persad AS, Koi F, Yamagoe S, Hashimoto K, et al. Novel missense mutation found in a Japanese patient with myeloperoxidase deficiency. Gene. 2004;327(2):195–200.Aratani Y, Koyama H, Nyui SI, Suzuki K, Kura F, Maeda N. Severe impairment in early host defense against Candida albicans in mice deficient in myeloperoxidase. Infect Immun. 1999;67(4):1828–36.Marchetti C, Patriarca P, Solero G Pietro, Baralle FE, Romano M. Genetic Characterization of Myeloperoxidase Deficiency in Italy. Hum Mutat. 2004;23(5):496–505Comino R, Davi E, Martı JC, Docente U, Gene D De. OBSTETRICIA Y GINECOLOG ´ La vulvovaginitis candidia. 2013;56(2):108–16. Available from: file:///C:/Users/Usuario/Downloads/S0304501312001641_S300_es.pdfPemán J, Quindós G. Aspectos actuales de las enfermedades invasoras causadas por Candida y otros hongos levaduriformes. Rev Iberoam Micol [Internet]. 2016;33(3):133–9. Available from: http://dx.doi.org/10.1016/j.riam.2015.10.001Pemán J, Quindós G. Aspectos actuales de las enfermedades invasoras causadas por Candida y otros hongos levaduriformes. Rev Iberoam Micol [Internet]. 2016;33(3):133–9. Available from: http://dx.doi.org/10.1016/j.riam.2015.10.001Kwak W, Han YH, Seol D, Kim H, Ahn H, Jeong M, et al. Complete Genome of Lactobacillus iners KY Using Flongle Provides Insight Into the Genetic Background of Optimal Adaption to Vaginal Econiche. Front Microbiol. 2020;11(May):1–10.Aldunate M, Srbinovski D, Hearps AC, Latham CF, Ramsland PA, Gugasyan R, et al. Antimicrobial and immune modulatory effects of lactic acid and short chain fatty acids produced by vaginal microbiota associated with eubiosis and bacterial vaginosis. Front Physiol. 2015;6(JUN):1–23Parolin C, Marangoni A, Laghi L, Foschi C, Palomino RAÑ, Calonghi N, et al. Isolation of vaginal lactobacilli and characterization of anti-candida activity. PLoS One. 2015;10(6):1– 17.Van De Wijgert JHHM, Borgdorff H, Verhelst R, Crucitti T, Francis S, Verstraelen H, et al. The vaginal microbiota: What have we learned after a decade of molecular characterization? PLoS One. 2014;9(8).Li J, McCormick J, Bocking A, Reid G. Importance of vaginal microbes in reproductive health. Reprod Sci. 2012;19(3):235–42.Li J, McCormick J, Bocking A, Reid G. Importance of vaginal microbes in reproductive health. Reprod Sci. 2012;19(3):235–42.Sha BE, Zariffard MR, Wang QJ, Chen HY, Bremer J, Cohen MH, et al. Female genitaltract HIV load correlates inversely with Lactobacillus species but positively with bacterial vaginosis and Mycoplasma hominis. J Infect Dis. 2005;191(1):25–32.Hayes R, Watson-Jones D, Celum C, Van De Wijgert J, Wasserheit J. Treatment of sexually transmitted infections for HIV prevention: End of the road or new beginning? Aids. 2010;24(SUPPL. 4):1–19Van De Wijgert JHHM, Morrison CS, Cornelisse PGA, Munjoma M, Moncada J, Awio P, et al. Bacterial vaginosis and vaginal yeast, but not vaginal cleansing, increase HIV-1 acquisition in African women. J Acquir Immune Defic Syndr. 2008;48(2):203–10.Jacobsen ID, Hube B. Candida albicans morphology: still in focus. Expert Rev Anti Infect Ther [Internet]. 2017;15(4):327–30. Available from: http://dx.doi.org/10.1080/14787210.2017.1290524Poulain D. Candida albicans, plasticity and pathogenesis. Crit Rev Microbiol. 2015;41(2):208–17.Szafranski-Schneider E, Swidergall M, Cottier F, Tielker D, Román E, Pla J, et al. Msb2 shedding protects Candida albicans against antimicrobial peptides. PLoS Pathog. 2012;8(2)Spettel K, Barousch W, Makristathis A, Zeller I, Nehr M, Selitsch B, et al. Analysis of antifungal resistance genes in Candida albicans and Candida glabrata using next generation sequencing. PLoS One. 2019;14(1):1–19.Gao J, Wang H, Li Z, Wong AHH, Wang YZ, Guo Y, et al. Candida albicans gains azole resistance by altering sphingolipid composition. Nat Commun [Internet]. 2018;9(1):1–14. Available from: http://dx.doi.org/10.1038/s41467-018-06944-1Silva S, Negri M, Henriques M, Oliveira R, Williams DW, Azeredo J. Candida glabrata, Candida parapsilosis and Candida tropicalis: Biology, epidemiology, pathogenicity and antifungal resistance. FEMS Microbiol Rev. 2012;36(2):288–305Ramos JT, Francisco L, Daoud Z. Infección fúngica invasora en niños: diferencias y analogías con el adulto. Rev Española Quimioter. 2016;29(1):59–65.Wu EY, Ehrlich L, Handly B, Frush DP, Rebecca H, Hill C, et al. Immunodeficiency Disorders : an Update. 2017;46(12):1630–44.Quindós G. Epidemiology of candidaemia and invasive candidiasis. A changing face. Rev Iberoam Micol [Internet]. 2014;31(1):42–8. Available from: http://dx.doi.org/10.1016/j.riam.2013.10.001Sanches JM, Giraldo PC, Bardin MG, Amaral R, Discacciati MG, Rossato L. Laboratorial Aspects of Cytolytic Vaginosis and Vulvovaginal Candidiasis as a Key for Accurate Diagnosis: A Pilot Study. Rev Bras Ginecol e Obstet. 2020;42(10):634–41.Uribarren-Berrueta T del NJ, Pineda-Murillo J, Cortés-Figueroa AÁ, Castañon Olivares LR. Candidosis vaginal. Segunda parte:diagnóstico, tratamiento y situación de algunos países latinoamericanos. Rev Médica Risaralda. 2017;23(1):159–66.Farr A, Effendy I, Frey Tirri B, Hof H, Mayser P, Petricevic L, et al. Guideline: Vulvovaginal candidosis (AWMF 015/072, level S2k). Mycoses. 2021;64(6):583–602.Pereira LC, Correia AF, da Silva ZDL, de Resende CN, Brandão F, Almeida RM, et al. Vulvovaginal candidiasis and current perspectives: new risk factors and laboratory diagnosis by using MALDI TOF for identifying species in primary infection and recurrence. Eur J Clin Microbiol Infect Dis. 2021;40(8):1681–93Candidosis vaginal - Diagnóstico y tratamiento - Mayo Clinic [Internet]. [cited 2021 Sep 5]. Available from: https://www.mayoclinic.org/es-es/diseases-conditions/yeastinfection/diagnosis-treatment/drc-20379004Colomer M. Candidiasis vulvovaginal Tratamiento tópico y oral. Actual científica Av Farm. 2009;28(1):68–9Pérez AM. Reacción en cadena de la polimerasa (Polymerase Chain Reaction, PCR). Repos Inst la Univ Politècnica València [Internet]. 2011;10. Available from: https://riunet.upv.es/bitstream/handle/10251/10700/Reacción en cadena de la polimerasa.pdfTamay de Dios L * Ibarra C,** Velasquillo C*. Fundamentos de la reacción en cadena de la polimerasa (PCR) y de la PCR en tiempo real. Medigraphic [Internet]. 2013 [cited 2021 Sep 5];2:70–8. Available from: www.medigraphic.org.mxhttp://www.medigraphic.com/ridwww.medigraphic.org.mxMárquez L, Serrato A, Cerritos R. Secuenciación de fragmentos de ADN. Herramientas Mol Apl en Ecol Asp teóricos y prácticos [Internet]. 2014;231–49. Available from: http://www2.inecc.gob.mx/publicaciones/libros/710/secuenciacion.pdfCarla LC, Fernando GC, María TC, Antonio OP, Emilia CM, Rafael CM. Aplicaciones de las técnicas de secuenciación masiva en la Microbiología Clínica [Internet]. 2021. 1–55 p. Available from: https://seimc.org/contenidos/documentoscientificos/procedimientosmicrobiologia/seimc-procedimiento71.pdfProducts A. Manual de funcionamiento BC 3000 plus Hematologia. Maquin Mont LTDA [Internet]. 2014;(7001):1–43. Available from: http://www.maquinariamontana.com/images/manuales/H616 618 620.pdf%0Ahttp://sistemamid.com/preview.php?a=6209NCBI. Cromosoma 17 de Homo sapiens, GRCh38.p13 Ensamblaje primario - Nucleótido - NCBI [Internet]. [cited 2021 Sep 6]. Available from: https://www.ncbi.nlm.nih.gov/nuccore/NC_000017.11?report=genbank&from=58269855& to=58280935&strand=trueCampos-Sanchez E, Martínez-Cano J, del Pino Molina L, López-Granados E, Cobaleda C. Epigenetic Deregulation in Human Primary Immunodeficiencies. Trends Immunol. 2019;40(1):49–65Capucine P, Bousfiha A, Jeddane L, Ailal F, Gaspar HB, Crow WA-H, et al. The 2017 IUIS Phenotypic Classification for Primary Immunodeficiencies. Lubr Eng. 1997;53(7):12–7Lanza F. Clinical manifestation of myeloperoxidase deficiency. J Mol Med. 1998;76(10):676–81.Carmel R, Watkins D, Rosenblatt DS. Chapter 10 – Megaloblastic Anemia. Nathan Oski’s Hematol Oncol Infancy Child. 8th Ed. 2015;308-343.e11.Cheng G, Salerno JC, Cao Z, Pagano PJ, Lambeth JD. Identification and characterization of VPO1, a new animal heme-containing Peroxidase.El-Ashram S, Al Nasr I, Suo X. Nucleic acid protocols: Extraction and optimization. Biotechnol Reports [Internet]. 2016;12:33–9. Available from: http://dx.doi.org/10.1016/j.btre.2016.10.001Metzler KD, Fuchs TA, Nauseef WM, Reumaux D, Roesler J, Schulze I, et al. Myeloperoxidase is required for neutrophil extracellular trap formation: Implications for innate immunity. Blood. 2011;117(3):953–9Quie PG, White JG, Holmes B, Good RA. In vitro bactericidal capacity of human polymorphonuclear leukocytes: diminished activity in chronic granulomatous disease of childhood. J Clin Invest. 1967;46(4):668–79Koziol-Montewka M, Magrys A, Paluch-Oles J, Bogut A, Buczynski K, Jablonka S. MPO and cytokines in the serum of cancer patients in the context of Candida colonization and infection. Immunol Invest. 2006;35(2):167–79.Vigezzi C, Riera FO, Rodriguez E, Icely PA, Miró MS, Figueredo CM, et al. Candidiasis invasora: un enfoque a la infección en el sistema nervioso central. Rev Argent Microbiol. 2021;53(2):171–8Serrato Díaz A, Flores Rentería L, Aportela Cortez J, Sierra Palacios E. PCR: reacción en cadena de la polimerasa. [cited 2021 Sep 5]; Available from: www.biologiaPolimorfismosDeficiencia de MieloperoxidasaAnálisis BioinformáticoPCRSecuenciaciónCandidiasis vulvovaginalCandida sppLICENSElicense.txtlicense.txttext/plain; charset=utf-814828https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5717/4/license.txt2f9959eaf5b71fae44bbf9ec84150c7aMD54open accessTEXTIdentificación de polimorfismos en el gen mieloperoxidasa y su papel en candidiasis vulvovaginal recurrente por Candida spp en pacientes de Bogotá DC.pdf.txtIdentificación de polimorfismos en el gen mieloperoxidasa y su papel en candidiasis vulvovaginal recurrente por Candida spp en pacientes de Bogotá DC.pdf.txtExtracted texttext/plain133808https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5717/5/Identificaci%c3%b3n%20de%20polimorfismos%20en%20el%20gen%20mieloperoxidasa%20y%20su%20papel%20en%20candidiasis%20vulvovaginal%20recurrente%20por%20Candida%20spp%20en%20pacientes%20de%20Bogot%c3%a1%20DC.pdf.txt801d2b9a1135c7975a16de8056561107MD55open access4. FORMATO IDENTIFICACIÓN TRABAJOS DE GRADO(1).pdf.txt4. FORMATO IDENTIFICACIÓN TRABAJOS DE GRADO(1).pdf.txtExtracted texttext/plain3243https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5717/7/4.%20FORMATO%20IDENTIFICACI%c3%93N%20TRABAJOS%20DE%20GRADO%281%29.pdf.txte05e93a1452a4c335cdf98fd5236eb38MD57metadata only accessCARTA DERECHOS DE AUTOR Montoya.pdf.txtCARTA DERECHOS DE AUTOR Montoya.pdf.txtExtracted texttext/plain1139https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5717/9/CARTA%20DERECHOS%20DE%20AUTOR%20Montoya.pdf.txt123eb2349f8a77bfa64c6bdf27fd125cMD59metadata only accessTHUMBNAILIdentificación de polimorfismos en el gen mieloperoxidasa y su papel en candidiasis vulvovaginal recurrente por Candida spp en pacientes de Bogotá DC.pdf.jpgIdentificación de polimorfismos en el gen mieloperoxidasa y su papel en candidiasis vulvovaginal recurrente por Candida spp en pacientes de Bogotá DC.pdf.jpgGenerated Thumbnailimage/jpeg6257https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5717/6/Identificaci%c3%b3n%20de%20polimorfismos%20en%20el%20gen%20mieloperoxidasa%20y%20su%20papel%20en%20candidiasis%20vulvovaginal%20recurrente%20por%20Candida%20spp%20en%20pacientes%20de%20Bogot%c3%a1%20DC.pdf.jpga7bea7b6fae6c8aec9b80437c67fb999MD56open access4. FORMATO IDENTIFICACIÓN TRABAJOS DE GRADO(1).pdf.jpg4. FORMATO IDENTIFICACIÓN TRABAJOS DE GRADO(1).pdf.jpgGenerated Thumbnailimage/jpeg13148https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5717/8/4.%20FORMATO%20IDENTIFICACI%c3%93N%20TRABAJOS%20DE%20GRADO%281%29.pdf.jpged029c2618f978e938d4434cdebd7e9eMD58metadata only accessCARTA DERECHOS DE AUTOR Montoya.pdf.jpgCARTA DERECHOS DE AUTOR Montoya.pdf.jpgGenerated Thumbnailimage/jpeg9341https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5717/10/CARTA%20DERECHOS%20DE%20AUTOR%20Montoya.pdf.jpg646cd9b36ffbf7f3c752978f84bd4224MD510metadata only accessORIGINALIdentificación de polimorfismos en el gen mieloperoxidasa y su papel en candidiasis vulvovaginal recurrente por Candida spp en pacientes de Bogotá DC.pdfIdentificación de polimorfismos en el gen mieloperoxidasa y su papel en candidiasis vulvovaginal recurrente por Candida spp en pacientes de Bogotá DC.pdfapplication/pdf5169255https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5717/11/Identificaci%c3%b3n%20de%20polimorfismos%20en%20el%20gen%20mieloperoxidasa%20y%20su%20papel%20en%20candidiasis%20vulvovaginal%20recurrente%20por%20Candida%20spp%20en%20pacientes%20de%20Bogot%c3%a1%20DC.pdf437853c9633a805b5cb1520eb9b14303MD511open accessCARTA DERECHOS DE AUTOR Montoya.pdfCARTA DERECHOS DE AUTOR Montoya.pdfapplication/pdf153134https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5717/12/CARTA%20DERECHOS%20DE%20AUTOR%20Montoya.pdf4b0e49e2b81b201c7954c7d506fbee93MD512metadata only access4. FORMATO IDENTIFICACIÓN TRABAJOS DE GRADO(1).pdf4. FORMATO IDENTIFICACIÓN TRABAJOS DE GRADO(1).pdfapplication/pdf196941https://repositorio.unicolmayor.edu.co/bitstream/unicolmayor/5717/13/4.%20FORMATO%20IDENTIFICACI%c3%93N%20TRABAJOS%20DE%20GRADO%281%29.pdfaa95c18a0f56d7461d118693f309aebcMD513metadata only accessunicolmayor/5717oai:repositorio.unicolmayor.edu.co:unicolmayor/57172024-05-10 11:12:20.328An 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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 |