ABSTRACT
In the present study, we investigated whether cellular damage, as demonstrated by lactate dehydrogenase (LDH) release in the human fallopian tube (FT) infected by Neisseria gonorrhoeae (Ngo), correlated with high levels of nitric oxide synthase (NOS) mRNA and enzyme activity. Infection with Ngo induced a significant increase (~35-fold) in mRNA transcripts of the inducible isoform of NOS. Paradoxically, a reduction in NOS enzyme activity was observed in infected cultures, suggesting that gonococcal infection possibly influences translation of iNOS mRNA to the enzyme. In addition, treatment with the NOS inhibitor TRIM did not prevent gonococcal-induced cellular damage. In contrast, the addition of the inhibitor L-NAME induced a 40 percent reduction in LDH release, which correlated with a ~50 percent reduction in gonococcal numbers. Moreover, treatment of normal FT explants with an exogenous NO donor, SNAP, did not induce significant cellular damage. Taken together, our data suggest that NO does not contribute to cellular damage during infection of the human FT with Neisseria gonorrhoeae.
Subject(s)
Female , Humans , Fallopian Tubes/microbiology , L-Lactate Dehydrogenase/metabolism , Neisseria gonorrhoeae/enzymology , Nitric Oxide Synthase/metabolism , Nitric Oxide/metabolism , RNA, Messenger/metabolism , Cells, Cultured , Fallopian Tubes/pathology , Time FactorsABSTRACT
Background: Infection of the Fallopian tubes (FT) by Neisseria gonorrhoeae (Ngo) can lead to acute salpingitis, an inflammatory condition resulting in damage primarily to the ciliated cells, with loss of ciliary activity and sloughing of the cells from the epithelium. Recently, we have shown that Ngo infection induced apoptosis in FT epithelium cells by a TNF-alpha dependent mechanism that could contribute to the cell and tissue damage observed in gonococcal salpingitis. Aim: To investigate the apoptosis-related genes expressed during apoptosis induction in cultured FT epithelial cells infected in vitro by Ngo. Materials and Methods: In the current study, we used cDNA macroarrays and real time PCR to identify and determine the expression levels of apoptosis related genes during the in vitro gonococci infection of FT epithelial cells. Results: Significant apoptosis was induced following infection with Ngo. Macroarray analysis identified the expression of multiple genes of the TNF receptor family (TNFRSF1B, -4, -6, -10A, -10B and -10D) and the Bcl-2 family (BAK1, BAX, BLK, HRK and MCL-1) without differences between controls and infected cells. This lack of difference was confirmed by RT-PCR of BAX, Bcl-2, TNFRS1A (TNFR-I) and TNFRSF1B (TNFR-II). Conclusion: Several genes related to apoptosis are expressed in primary cultures of epithelial cells of the human Fallopian tube. Infection with Ngo induces apoptosis without changes in the pattern of gene expression of several apoptosis-related genes. Results strongly suggest that Ngo regulates apoptosis in the FT by post-transcriptional mechanisms that need to be further addressed.
Subject(s)
Female , Humans , Apoptosis/genetics , Epithelial Cells/microbiology , Fallopian Tubes/microbiology , Neisseria gonorrhoeae/physiology , Salpingitis/microbiology , Cells, Cultured , Epithelial Cells/pathology , Fallopian Tubes/pathology , Gene Expression Regulation , Oligonucleotide Array Sequence Analysis , /metabolism , Reverse Transcriptase Polymerase Chain Reaction , Receptors, Tumor Necrosis Factor/metabolism , Salpingitis/pathologyABSTRACT
Introducción. Desde 1948, la exenteración pélvica ha sido la alternativa de tratamiento en el rescate para pacientes con recurrencia por cáncer en la pelvis, sobre todo de tipo ginecológico. El procedimiento original ha sido modificado infinidad de ocasiones en un intento por mejorar el estado general y la calidad de vida de las pacientes sometidas a esta intervención. Objetivo. Efectuamos una revisión de la literatura y transmitimos la evaluación sobre el abordaje integral de las pacientes con recurrencia por cáncer ginecológico, con especial énfasis en el manejo perioperatorio estándar de las pacientes candidatas a esta intervención. Material y métodos. Revisión de la literatura, y análisis crítico de los departamentos involucrados, en el manejo perioperatorio de estas pacientes. Se describen con detalle desde el proceso de selección de las mujeres candidatas, rutas de manejo, monitoreo, evaluaciones preoperatorias, técnica quirúrgica más comúnmente empleada en el Instituto Nacional de Cancerología, hasta los cuidados posoperatorios y seguimiento de las pacientes. Resultados. Resultados. La selección meticulosa de las pacientes candidatas al procedimiento se inicia desde la primera entrevista con evaluación del entorno de la pacientes, soporte familiar, factibilidad económica y decision final por parte de la paciente. Participación de un equipo quirúrgico y de enfermería, multidisplinario, familiarizado con el procedimiento y colaborando sincrónicamente en las fases exenterativa y de reconstrucción. Apego y compromiso en la vigilancia estrecha del posoperatorio y evolución a corto, mediano y largo plazo de las pacientes. Conclusiones. El éxito de este tipo de procedimiento requiere pacientes bien seleccionadas, en centro oncológicos de experiencia, participación multidisciplinaria durante la resección quirúrgica, pero sobre todo del compromiso real por parte del equipo quirúrgico y de enfermería durante el manejo posoperatorio