Project description:Bacterial vaginosis (BV) is the most common vaginal infection in women of reproductive age and has been associated with serious health complications, mainly in pregnant women. It is characterized by a decrease in the number of Lactobacillus species in the healthy vaginal microbiota and an overgrowth of strict and facultative anaerobic bacteria that develop a polymicrobial biofilm. Despite over 60 years of research investigating BV, its etiology is not fully understood. Gardnerella spp. is a crucial microorganism that contributes to the formation of the biofilm and the development of BV, but the role of other BV-associated bacteria is not clear. Nevertheless, Fannyhessea vaginae (previously known as Atopobium vaginae) is a highly specific species for BV, and co-colonization with Gardnerella is thought to be a very specific diagnostic marker. The diagnosis of BV still presents some limitations, since currently used methods often fail to accurately detect BV. This work aims to develop a novel peptide nucleic acid (PNA) probe targeting F. vaginae. This probe was further validated in a multiplex assay, which included a Gardnerella-specific PNA probe, as a possible method for diagnosis of BV, and was compared with quantification by qPCR. The new PNA probe showed excellent sensitivity and specificity and could discriminate F. vaginae-Gardnerella biofilms, confirming the potential to be used for the detection of BV-associated pathogens.
Project description:Gene expression in the neonatal uterus and vagina from transformation related protein 63 (Trp63) conditional heterozygous and null mice was analyzed in order to identify upstream and downstream of Trp63 in the developing vagina. Differential gene expression between vaginal (Trp63 positive) and uterine (Trp63 negative) tissues should reflect both the upstream and downstream of Trp63 expression, whereas the comparison between conditional knockout and heterozygous vaginae should identify what is downstream of Trp63. Thus, the three-way comparison should identify what is upstream of Trp63. Total RNA was extracted from the Trp63 conditional heterozygous (Trp63 flox/wt ; Pax2-cre) uteri and vaginae, and Trp63 conditional knockout (Trp63 flox/floxt ; Pax2-cre) vaginae. Total 12 samples (3 different genotype/tissue groups x 4 independent samples each).