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1.
ARS med. (Santiago, En línea) ; 45(3): 29-35, sept. 30, 2020.
Article in Spanish | LILACS | ID: biblio-1255300

ABSTRACT

Introducción: La enfermedad celíaca (EC) es una patología autoinmune, que se desarrolla a cualquier edad en personas genéticamentesusceptibles, y cuyo órgano diana principal es el intestino delgado. La diversidad en las formas de presentación actualmente conocidas implica un desafío permanente para el laboratorio, que debe ofrecer nuevas técnicas, cada vez más sensibles y específicas, para detectar de manera eficiente los autoanticuerpos específicos para el diagnóstico y seguimiento de estos pacientes. Nuestro objetivo fue evaluar la sensibilidad y especificidad de un nuevo antígeno para la detección de anticuerpos anti-transglutaminasa compuesto por transglutaminasa tisular unida covalentemente a péptidos deamidados de gliadina (neo-epítope) en pacientes con sospecha de EC, con biopsia duodenal como gold standard. Como objetivo secundario nos propusimos evaluar la sensibilidad y especificidad del antígeno convencional, transglutaminasa humana recombinante, para el mismo grupo de estudio. Metodología: Se realizó un estudio prospectivo, analizando muestras de pacientes con sospecha de EC o en seguimiento de dicha patología, en las que se estudiaron los anticuerpos anti-transglutaminasa con antígeno neo- epítope, y con antígeno transglutaminasa recombinante humana. Se determinó sensibilidad, especificidad, VPP, VPN y coeficiente de concordancia Kappa. Resultados: Se procesaron 56 muestras, incluidas en un período de 5 meses. La sensibilidad (100%) y especificidad (92,3%) obtenidas con la técnica de neo-epítope, en relación a la biopsia (gold standard), fue mayor que con la técnica transglutaminasa humana recombinante (88,3% y 78,9% respectivamente). La técnica con neo-epítope proporcionó un menor número de resultados en la "zona de indeterminación". Conclusiones: Nuestros resultados concuerdan con otros autores, ya que neo-epítope detecta con mayor sensibilidad y especificidad aquellos pacientes con diferente situación de presentación y transgresores de la dieta libre de gluten, quienes pueden presentar serología negativa o débilmente positiva con transglutamiasa humana recombinante. La nueva técnica neo-epítope constituiría una mejor herramienta para la pesquisa diagnóstica y de seguimiento en pacientes con EC.


Introduction: Celiac disease (CD) is an autoimmune disease, which develops at any age in genetically susceptible people, and whose main target organ is the small intestine. The diversity in the currently known forms of presentation implies a permanent challenge for the laboratory, which must offer new techniques, increasingly sensitive and specific, to efficiently detect the specific autoantibodies that collaborate in the diagnosis and follow-up of these patients. Our main objective was to evaluate the sensitivity and specificity of a new sensitizing antigen for the detection of anti-transglutaminase antibodies composed of tissue transglutaminase covalently linked to deamidated gliadin peptides (neo-epitope) in patients with suspected CD, with duodenal biopsy as the gold standard. As a secondary objective, we set out to evaluate the sensitivity and specificity of the conventional antigen, recombinant human transglutaminase, for the same study group. Methodology: A prospective study was carried out, including samples from patients with suspected CD or in follow-up of said pathology, in which anti-transglutaminase antibodies were studied with neo-epitope antigen, and with human re-combinant transglutaminase antigen. Sensitivity, specificity, PPV, NPV and Kappa coefficient of concordance were determined. Results: 56 samples were processed, included in a period of 5 months. The sensitivity and specificity obtained with the neo-epitope technique (S: 100% - E: 92.3%), in relation to the biopsy (gold standard), was higher than with the recombinant human transglutaminase technique (S: 88.3% - E: 78.9%). The neo-epitope technique provided fewer results in the "zone of indeterminacy". Conclusions: Our results agree with other authors, since the neo-epitope detects with greater sensitivity and specificity those patients with different presentation situations and transgressors of the gluten-free diet, who can present negative or weakly positive serology with recombinant human transglutaminase. The new neo-epitope technique would constitute a better tool for diagnostic and follow-up research in patients with CD


Subject(s)
Autoantibodies , Celiac Disease , Prospective Studies , Patients , Autoimmunity , Antibodies
2.
Biomédica (Bogotá) ; 39(3): 547-560, jul.-set. 2019. graf
Article in Spanish | LILACS | ID: biblio-1038814

ABSTRACT

Resumen Introducción. La leptina es una hormona secretada por los adipocitos que se ha relacionado con el proceso de la transición de epitelio a mesénquima (Epithelial- Mesenchymal Transition, EMT). Promueve la migración e invasión de las células del epitelio mamario mediante la activación de las cinasas FAK y Src, un complejo regulador de vías de señalización que favorecen la expresión de las proteínas relacionadas con la formación de estructuras proteolíticas implicadas en la invasión y progresión del cáncer. Recientemente, se ha descrito que la sobreexpresión y activación de la proteína Hic-5 durante el mencionado proceso de transición, favorece la formación de los puntos de actina (indicativa de la formación y funcionalidad de los invadopodios), lo cual promueve la degradación local de los componentes de la matriz extracelular y la metástasis del cáncer. Objetivos. Evaluar el papel de las cinasas FAK y Src sobre la expresión y localización subcelular de Hic-5 y la formación de puntos de actina inducida por la leptina en la línea celular MCF10A de epitelio mamario no tumoral. Materiales y métodos. Se utilizaron los inhibidores específicos de la FAK (PF-573228) y la Src (PP2) para evaluar el papel de ambas cinasas en los niveles de expresión y localización subcelular de la proteína Hic-5 mediante Western blot e inmunofluorescencia, así como la formación de puntos de actina mediante la tinción con faloidina-TRITC en células MCF10A estimuladas con leptina. Resultados. La leptina indujo el incremento en la expresión de Hic-5 y la formación de puntos de actina. El tratamiento previo con los inhibidores de las cinasas FAK (PF-573228) y Src (PP2), promovió la disminución en la expresión de Hic-5 y de los puntos de actina en la línea celular MCF10A de epitelio mamario no tumoral. Conclusión. La leptina indujo la expresión y la localización perinuclear de Hic-5 y la formación de puntos de actina mediante un mecanismo dependiente de la actividad de las cinasas FAK y Src en las células MCF10A.


Abstract Introduction: Leptin is a hormone secreted by adipocytes that has been associated with the epithelial-mesenchymal transition (EMT). Additionally, leptin promotes the migration and invasion of mammary epithelial cells through the activation of FAK and Src kinases, which are part of a regulatory complex of signaling pathways that promotes the expression of proteins related to the formation of proteolytic structures involved in the invasion and progression of cancer. Recently, overexpression and activation of Hic-5 during the EMT have been shown to induce the formation of actin puncta; these structures are indicative of the formation and functionality of invadopodia, which promote the local degradation of extracellular matrix components and cancer metastasis. Objective: To evaluate the role of FAK and Src kinases in the expression of Hic-5 during the epithelial-mesenchymal transition induced by leptin in MCF10A cells. Materials and methods: We used specific inhibitors of FAK (PF-573228) and Src (PP2) to evaluate Hic-5 expression and subcellular localization by Western blot and immunofluorescence assays and to investigate the formation of actin puncta by epifluorescence in MCF10A cells stimulated with leptin. Results: Leptin induced an increase in Hic-5 expression and the formation of actin puncta. Pretreatment with inhibitors of FAK (PF-573228) and Src (PP2) promoted a decrease in Hic-5 expression and actin puncta formation in the non-tumorigenic mammary epithelial cell line MCF10A. Conclusion: In MCF10A cells, leptin-induced Hic-5 expression and perinuclear localization, as well as the formation of actin puncta through a mechanism dependent on the kinase activity of FAK and Src.


Subject(s)
Humans , src-Family Kinases/physiology , Leptin/pharmacology , Intracellular Signaling Peptides and Proteins/metabolism , Fanconi Anemia Complementation Group C Protein/physiology , Epithelial-Mesenchymal Transition/drug effects , LIM Domain Proteins/metabolism , Pyrimidines/pharmacology , Sulfones/pharmacology , Signal Transduction , Cell Line , Actins , Quinolones/pharmacology , src-Family Kinases/antagonists & inhibitors , Epithelial Cells/drug effects , Epithelial Cells/metabolism , Fanconi Anemia Complementation Group C Protein/antagonists & inhibitors , Epithelial-Mesenchymal Transition/physiology , Neoplasm Invasiveness
3.
Rev. bras. ter. intensiva ; 19(3): 354-356, jul.-set. 2007. ilus, tab
Article in Portuguese | LILACS | ID: lil-470948

ABSTRACT

JUSTIFICATIVA E OBJETIVOS: A endocardite é rara e grave complicação de brucelose humana. O diagnóstico é suspeito em casos de endocardite sem resposta ao tratamento comum e é confirmado com sorologia com teste de ELISA maior que 1:160. O tratamento na maioria dos casos é cirúrgico, associado a antibioticoterapia prolongada. Alguns casos são resolvidos com tratamento clínico, que inclui doxiciclina, rifampicina, ciproproxacina, gentamicina e tetraciclina. O objetivo deste estudo foi relatar um caso raro de endocardite por brucelose. RELATO DO CASO: Paciente do sexo masculino, 51 anos, trabalhador rural, admitido com quadro de febre e emagrecimento. O ecocardiograma mostrou espessamento e vegetação de válvula aórtica. Uma hemocultura foi positiva para Staphylococcus epidermidis. O tratamento foi iniciado no dia da internação com penicilina cristalina, associada com garamicina, sem melhora em três semanas. Foi suspeitada endocardite por brucelose, colhida sorologia e iniciado o tratamento. Os antibióticos usados foram rifampicina e ciprofloxacin associado à vancomicina, devido o resultado da primeira hemocultura. A sorologia de aglutinação para brucelose foi positiva com resultado de 1:360. O paciente melhorou e recebeu alta da UTI, hemodinamicamente estável e em uso de ciprofloxacin e gentamicina. CONCLUSÕES: A endocardite por brucelose não é comum, mas deve ser sempre lembrada quando o tratamento convencional de endocardite não tem boa resposta, principalmente nos pacientes com possível contato com animais e derivados de leite.


BACKGROUND AND OBJECTIVES: Endocarditis is a rare and serious complication of human brucellosis. The diagnosis is suspected in cases of endocarditis without response to conservative antibiotic treatment and it is confirmed with enzyme-linked immunosorbent assay (ELISA) test, titers being higher than 1:160. The treatment is usually a surgery, followed with antibiotics for long period of time. Some cases can be cured with antibiotic treatment only, with antibiotics such as doxiciclin, rifampicin, ciprofloxacin, gentamicin and tetracycline. We present a case report of a patient with brucellose endocarditis. CASE REPORT: Fifty one year old male patient, a farmer, was admitted with clinical history of fever and weight loss. Echocardiography showed thickening and vegetation on the aortic valve and blood culture was positive for Staphylococcus epidermidis. The treatment with crystal penicillin and garamycin was initiated with no improval during three weeks. Endocarditis caused by human brucellosis was suspected and a new treatment with rifampicin and ciprofloxacin, associated with vancomycin because of the first blood culture, was initiated. Agglutination sorology was positive for brucellosis, with titers of 1:360. The patient got better with new treatment and was dismissed from the intensive care unit clinically stable, taking ciprofloxacin and gentamicin. CONCLUSIONS: Endocarditis caused by human brucellosis is rare; however it should always be considered when conservative antibiotic treatment fails, especially in patients that have contact with animals and dairy products.


Subject(s)
Humans , Male , Middle Aged , Brucellosis , Endocarditis, Bacterial
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