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
Mycoplasma canadense, a clinical isolate from milk of a mastitic buffalo, was experimentally tested for its pathogenic potential in hamster tracheal ring and rabbit fallopian tube explant organ cultures (in vitro) and rat and rabbit mammary gland (in vivo) models. The activity percentage reduction in M. canadense infected hamster tracheal rings was 99.1% in comparison to 16.4% in control rings. Mycoplasma canadense, also induced complete ciliostasis at 11-day post-infection in rabbit fallopian tube explants. Histopathological lesions in these infected organ cultures were loss of cilia, desquamation or denudation of epithelium, infiltration of inflammatory cells and proliferation of macrophages as well as oedema in lamina propria. At the end of the experiments, M. canadense organisms were reisolated in pure colonies from the infected but not the control organ cultures. In the rat and rabbit mammary glands, M. canadense organisms persisted upto 6-day and 7-day postinfection, respectively and caused histopathological changes suggestive of subacute to chronic mastitis during the experimental period. The results indicate that the tested M. canadense clinical isolate was virulent.
Subject(s)
Animals , Cattle , Cattle Diseases/microbiology , Cell Division , Cilia , Cricetinae , Edema , Epithelium , Fallopian Tubes/microbiology , Female , Macrophages , Mastitis/microbiology , Milk/microbiology , Mycoplasma/isolation & purification , Mycoplasma Infections/microbiology , Organ Culture Techniques , Rabbits , Rats , Trachea/microbiologyABSTRACT
A total of 100 cases were studied. Fallopian tubes removed at operation constituted a very good material to study the infections of upper genital tract. Along with fallopian tubes, HVS from same patients were cultured both for aerobic and anaerobic organisms. Bacteriological analysis of 100 cases of fallopian tubes and same number of HVS showed bacterial growth in 30 cases (30%) whereas in HVS samples growth was obtained in 50 cases (50%). In 66.67% of cases same bacteria were isolated both from HVS and fallopian tubes which strongly favours the theory of ascending infection. Out of all antibiotics ciprofloxacin showed the widest spectrum of activity (94.20%). All anaerobes were sensitive to metronidazole.
Subject(s)
Bacteria/isolation & purification , Drug Resistance, Microbial , Drug Resistance, Multiple , Fallopian Tubes/microbiology , Female , Humans , Microbial Sensitivity Tests , Pelvic Inflammatory Disease/diagnosis , Species Specificity , Vaginal SmearsABSTRACT
La flora microbiana abdominal y cervical fue estudiada en un grupo de 29 mujeres infértiles asintomáticas sometidas a laparoscopia. Se practicaron cultivos para gérmenes aerobios, anaerobios, neisseria gonorrhoeae, mycoplasma, ureaplasma urealyticum y chlamydia trachomatis. Se establecieron dos grupos de pacientes de acuerdo a la presencia o ausencia de factor tuboperitoneal. No se demostró relación entre la existencia de gérmenes en cervix y/o abdomen y factor tuboperitoneal. El antecedente de uso de dispositivo intrauterino (DIU) se asoció con la presencia de alteraciones tuboperitoneales
Subject(s)
Humans , Female , Adult , Cervix Uteri/microbiology , Fallopian Tubes/microbiology , Infertility, Female/microbiology , Bacteria, Aerobic/isolation & purification , Bacteria, Anaerobic/isolation & purification , Chlamydia trachomatis/isolation & purification , Infertility, Female/etiology , Mycoplasma/isolation & purification , Ureaplasma/isolation & purificationABSTRACT
Mollicutes (10) belonging to Mycoplasma and Acholeplasma isolated from various reproductive disorders were tested in rabbit fallopian tube (FT) organ culture. Parameter for describing pathogenic status of Mollicutes in rabbit FT organ culture included multiplication of organisms, and its effect on ciliary activity along with histopathological changes in FT explants. M. mycoides (LC, Y-Goat), M. bovoculi, M. bovigenitalium, Mycoplasma sp. and A. oculi were categorized as pathogenic; A. axanthum and A. laidlawii as mildly pathogenic; and M. bovis, M. arginini. and A. granularum, as nonpathogenic to rabbit FT organ culture. Thus, rabbit FT organ culture is recommended for use as a suitable and economical in vitro model to assess the pathogenicity of Mollicutes of reproductive tract origin.