ABSTRACT
AANTEDENTES: N. gonorrhoeae es una de las principales causas de uretritis, prostatitis y epididimitis en hombres. En el tracto urogenital masculino, esta bacteria o los factores solubles producto de su metabolismo pueden interactuar con los espermatozoides y alterar su calidad. OBJETIVOS: Determinar el efecto de la incubación in vitro de N. gonorrhoeae y los factores solubles producto de su metabolismo sobre la calidad espermática y evaluar la interacción bacteria-espermatozoide. MÉTODO: Se realizó incubación de una concentración 0,5 McFarland de N. gonorrhoeae y de los factores solubles producto de su metabolismo bacteriano con muestras de semen de voluntarios aparentemente sanos. Se cuantificaron los parámetros espermáticos convencionales (movilidad y viabilidad) y funcionales (potencial de membrana mitocondrial, integridad y lipoperoxidación de la membrana espermática, detección de especies reactivas del oxígeno, integridad de la cromatina y expresión de Anexina V), empleando microscopía y citometría de flujo, respectivamente. Para evaluar la interacción de N. gonorrhoeae con los espermatozoides humanos se realizaron extendidos en placas para su observación al microscopio. RESULTADOS: N. gonorrhoeae puede unirse al espermatozoide y disminuir la viabilidad espermática luego de 1,5 horas de incubación con los espermatozoides humanos (84,5% vs 66,5%, p<0,05), sin afectar los parámetros espermáticos funcionales. CONCLUSIÓN: N. gonorrhoeae interactúa con los espermatozoides humanos afectando la viabilidad espermática.
BACKGROUND: N. gonorrhoeae is a major cause of urethritis, prostatitis and epididymitis in men. In the male urogenital tract, the bacteria or soluble products of their metabolism may interact with sperm, and alter their quality. AIMS: To determine the in vitro effect of incubation of N. gonorrhoeae and soluble products of their metabolism on sperm quality and assess the bacteria-sperm interaction. METHODS: Soluble products of bacterial metabolism and 0.5 McFarland concentration of N. gonorrhoeae were incubated with semen samples from healthy volunteers. Conventional (motility and viability) and functional sperm parameters (potential of mitochondrial membrane integrity and sperm membrane lipid peroxidation, detection of reactive oxygen species, chromatin integrity and expression of annexin V) were quantified using microscopy and cytometry flow, respectively. To assess the interaction of N. gonorrhoeae to human sperm microscopic observation was performed. RESULTS: N. gonorrhoeae can join the sperm and decreases sperm viability after 1.5 hours of incubation with human sperm (84.5% vs 66.5%, p<0.05), without affecting the functional sperm parameters. CONCLUSION: N. gonorrhoeae interacts with human sperm affecting sperm viability.
Subject(s)
Humans , Male , Solubility , Spermatozoa/microbiology , Semen Analysis , Neisseria gonorrhoeae/metabolism , Sperm Motility/physiology , In Vitro Techniques , Flow Cytometry , Neisseria gonorrhoeae/physiologyABSTRACT
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
The study of the complement-outer bacterial membrane interactions is gaining enthusiasm for applying a continous development of more specialized techniques. In this paper a novel flow cytometry technique shows that highly fluorescent lucifer yellow-stained Neisseriae gonorrhoeae when exposed to the redox reaction of p-nitro blue chloride tetrazolium (NBT), as formazan precipitation takes place, a shift to lesser fluorescent channels of the histogram population occurs. That effect is labeled NBT-laser beam quenching (NBT-LBQ). A significant difference by Kolmogorov-Smirnov summation curve analysis is found between complement heat-inactivated and its counterpart assessed with normal complement microorganisms. The operation of this NBT-LBQ effect by microbial flow cytometry casts an interesting potential for the evaluation of the outer membrane-complement-interaction of serum sensitive microorganisms
Subject(s)
Flow Cytometry/methods , Complement System Proteins/physiology , Escherichia coli/physiology , Fluorescence , Neisseria gonorrhoeae/physiology , Nitroblue Tetrazolium , Data Interpretation, StatisticalABSTRACT
Se presenta un estudio in vitro de la migración de Neisseria gonorrheae adherido al espermatozoide humano através del moco cervical estrogénico. Se describen los tipos de contactos que caracterizan esta adherencia celular, basado en la observación de las muestras con microscopía electónica. Esta interacción celular podría tambíen explicar la génesis de los procesos inflamatorios pelvianos de origen gonocícito, mediante el transporte activo del agente etiológico
Subject(s)
Humans , Male , Female , In Vitro Techniques , Neisseria gonorrhoeae/physiology , Pelvic Inflammatory Disease/etiology , Spermatozoa/ultrastructure , Bacterial AdhesionABSTRACT
De acuerdo a Kellogg, Neisseria gonorrohoeae en cuatro (4) tipos. Los dos primeros son virulentos (T1 y T2). Se responsabiliza de este efecto a factores de adhesión (fimbria o pili) quienes permiten la colonización. En el presente estudio se pone en evidencia dichos factores por medio de reacción de hemoglutinación. Se trabajó con 89 cepas de aislamientos recientes obtenidas de pacientes con diagnóstico clínico de gonorrea; se utilizó la microtécnica en portaobjetos descripta por Koransky y colaboradores. Los ensayos se realizaron con eritrócitos pertenecientes a distintas especies y tipos sanguíneos. Se demonstró que los resultados no se modifican cuando se varía la procedencia o tipo de glóbulos rojos usados. Paralelamente, con sueros humanos antigonocócicos se efectuó la reacción de inhibición de hemoglutinación (IHA) frente a las cepas aisladas. Se confirma con estas pruebas la presencia de pili
Subject(s)
Humans , Bacterial Adhesion , Erythrocytes , Neisseria gonorrhoeae/physiology , Hemagglutination Tests , Neisseria gonorrhoeae/isolation & purificationABSTRACT
O artigo traz informaçöes sobre Gonorréia deste o histórico da doença, definiçäo, tipos clínicos até o diagnóstico e seu tratamento.