The role of 17β-estrogen in Escherichia coli adhesion on human vaginal epithelial cells via FAK phosphorylation

Estrogen, the predominant sex hormone, has been found to be related to the occurrence of vaginal infectious diseases. However, its role in the occurrence and development of bacterial vaginitis caused by Escherichia coli is still unclear. The objective of this study was to investigate the role of 17β-estrogen in E. coli adhesion on human vaginal epithelial cells. The vaginal epithelial cell line, VK2/E6E7, was used to study the molecular events induced by estrogen between E. coli and cells. An adhesion study was performed to evaluate the involvement of the estrogen-dependent focal adhesion kinase (FAK) activation with cell adhesion. The phosphorylation status of FAK and estrogen receptor α (ERα) upon estrogen challenge was assessed by Western blotting. Specific inhibitors for ERα were used to validate the involvement of ERα-FAK signaling cascade. The results showed that, following the stimulation with 1000 nM estrogen for 48 h, a transient activation of ERα and FAK was observed, as well as the increased average number of E. coli adhering to vaginal epithelial cell. In addition, estrogen-induced activation of ERa and FAK was inhibited by the specific inhibitor of ERα, especially when the inhibitor reached a 10 μM concentration and acted for 1 h, and a decrease in the number of adherent E. coli was observed simultaneously. However, this inhibitory effect diminished as the concentration of estrogen increased. In conclusion, FAK and ERα signaling cascades were assosiated with the increasing E. coli adherence to vaginal epithelial cells, which was promoted by a certain concentration of estrogen.

The RD2 pathogenicity island modifies the disease potential of the group A Streptococcus.

Serotype M28 isolates of the group A Streptococcus (GAS; Streptococcus pyogenes) are non-randomly associated with cases of puerperal sepsis, a potentially life-threatening infection that can occur in women following childbirth. Previously, we discovered that the 36.3 kb RD2 pathogenicity island, which is present in serotype M28 isolates but lacking from most other isolates, promotes the ability of M28 GAS to colonize the female reproductive tract. Here, we performed a gain-of-function study in which we introduced RD2 into representative serotype M1, M49, and M59 isolates and assessed the phenotypic consequences of RD2 acquisition. All RD2-containing derivatives colonized a higher percentage of mice, and at higher colony-forming-unit levels, than did the parental isolates in a mouse vaginal colonization model. However, for two additional phenotypes, survival in heparinized whole human blood and adherence to two human vaginal epithelial cell lines, there were serotype-specific differences to RD2-acquisition. Using transcriptomic comparisons, we identified that such differences may be a consequence of RD2 altering the abundance of transcripts from select core genome genes along serotype-specific lines. Our study is the first that interrogates RD2 function in GAS serotypes other than M28 isolates, shedding light on variability in the phenotypic consequences of RD2 acquisition, and informing on why this mobile genetic element is not ubiquitous in the GAS population.

Personal and Clinical Vaginal Lubricants: Impact on Local Vaginal Microenvironment and Implications for Epithelial Cell Host Response and Barrier Function

Vaginal lubricants are widely used both in-clinic and for personal use. Here, we employed monolayer and 3-dimensional vaginal epithelial cell models to show that select hyperosmolar lubricants induce cytotoxicity, reduce cell viability, and alter barrier and inflammatory targets.

Ultrasonographic and vaginal cytological diagnostics of the Queen

Ultrasonography is a diagnostic method to image the conditions of reproductive organs and it could be supported by vaginal cytology to identify the activities of the ovaries by the types of vaginal exfoliate cells. The aims of this study was to observe reproduction organ through ultrasonography with supportive diagnostic with vaginal cytological assessment. A total of 10 individual queens were used in this study and then grouped into intact group (n=5) and spayed (ovariohysterectomy) group (n=5) based on the anamneses or their medical history. The vagina, cervix, uterus body and horns, and ovaries were imaged and measured by ultrasound. Vagina, uterine body and horn seem as pipe-like structures with hyperechoic outer lines. The lumen in uterine body and horn seem as a hyperechoic structure. The ovaries seem as round- or oval-shaped structures with anechoic follicles. The corpus luteal has thick wall and seen as anechoic in its centre part. The corpus albicans seems as a hyperechoic structure. The vagina of spayed queens seemed more corrugated than those intact queens. The cervix is seen as a hyperechoic structure linking the vagina and uterine body. Exfoliate vaginal epithelial cell types were then also be identified and counted on each queens. The results of vaginal cytology showed that proestrus occured in 3 intact queens, late metestrus in 1 intact and 3 spayed queens, anestrus in 1 spayed queen, and unidentifiable estrus stage in 1 intact and 1 spayed queens. Moreover, the morphology of cervix and uterine was affected by the activity of ovary.

Saccharomyces cerevisiae-based probiotic as novel anti-fungal and anti-inflammatory agent for therapy of vaginal candidiasis

Previously we demonstrated that the treatment with live Saccharomyces cerevisiae exerts beneficial therapeutic effects against vaginal candidiasis. Here, we address potential mechanisms particularly examining the probiotic capacity to modulate both fungus and host-related factors. We show that the S. cerevisiae-based probiotic markedly affects the expression of virulence traits of Candida albicans such as aspartyl proteinases (SAPs) as well as hyphae-associated proteins Hwp1 and Ece1 in the vaginal cavity. On the host side, the probiotic suppression of the influx of neutrophils caused by the fungus into the vaginas of the mice is likely related to: (1) lower production of interleukin-8; and (2) inhibition of SAPs expression. However, these neutrophils displayed reactive oxygen species hyperproduction and increased killing activity as compared to the neutrophils of placebo-treated mice. There was no evidence of any cytotoxic effect by the probiotic, either when used in vivo on vaginal epithelial cell and organ architecture, or in in vitro in human vaginal epithelium. Inactivated yeast cells did not affect any of the factors above. In summary, the data suggest that the beneficial effect exerted by this S. cerevisiae-based probiotic is the result of its interference with the expression of fungus virulence factors coupled with the modulation of the inflammatory response of the host.