Vaginal Epithelium Research Articles

The role of probiotics in maintaining vaginal ecosystem

A disturbed vaginal ecosystem is characterized by reduced or non-existing lactobacillary flora and damaged vaginal epithelium. It may not always be apparent whether alterations of the vaginal epithelium or pathogenic microorganisms are the primary cause of a disturbed vaginal ecosystem. Estriol plays a key role in developing and maintaining a healthy vaginal epithelium, stimulating cell growth and differentiation, increases epithelial thickness, and also promotes glycogen synthesis, which is required to maintain an acidic environment. Estriol deficiency results in the vulvovaginal atrophy and the development of atrophic vaginitis. In turn, Lactobacillus acidophilus maintains a healthy microbiome, protecting against infections and creating an acidic environment unfavourable for the growth of pathogens. In recent years, an increasing interest has been developed in probiotics with the belief that probiotics could be able to restore the vaginal microbiota health. A review of scientific articles on the use of an ultra-low-dose vaginal estriol 0.03 mg in combination with Lactobacillus acidophilus to normalize the vaginal microbiota and treat vaginal infections has been conducted. An analysis of published clinical studies and meta-analyses on the use of probiotics in vaginal infections showed the mechanisms of action of probiotics, their biological role and clinical effects. The integration of probiotics into the treatment and prevention of bacterial vaginosis is growing more urgent. The results of numerous studies and meta-analyses confirmed their high efficacy and safety. The supplementation of standard antibiotic therapy with probiotics improves cure rates, restores normal vaginal microflora and reduces the risk of relapse. Therefore, further studies on the use of probiotics to develop more effective and personalized treatment strategies providing long-term results are needed. The publication of new evidence will make it possible to introduce best practices into clinical practice and ensure a healthier future for patients worldwide.

The diagnostic role of Lactobacillus spp. as representatives of the normal microbiome of the lower urogenital tract

Currently, scientific knowledge about various Lactobacillus species, which are representatives of the resident flora of the vagina, is actively expanding. We know, lactobacilli are an integral part of the female genital tract. Currently, about 20 different types of lactobacilli are known. Most of the lactobacilli already studied contribute to maintaining vaginal health by providing protection from pathogenic microorganisms, the development of dysbiosis, pregnancy complications, including premature birth. The protective function is provided primarily by the formation of Dand L-forms of lactic acid, which maintains an optimal acidic pH, from the glycogen of the vaginal epithelium. Lactobacillus spp. also prevent adhesion and invasion of pathogens into epithelial cells, synthesize bacteriocins, and inhibit the release of proinflammatory cytokines. However, to date, there is already evidence that not all representatives of lactobacilli have a positive effect on the health of the female body. Thus, patients with the CSTIII morphotype, dominated by L. iners, are especially predisposed to the development of vaginal dysbiosis, and the dominance of L. acidophilus in patients is probably associated with infertility. Currently, there is no reliable evidence of the positive and/or negative effects of many lactobacilli on the female body, and existing studies on a number of bacteria are limited and contradictory. In this regard, it is relevant to further study the characteristics and properties of Lactobacillus spp., colonizing the vagina, for a more accurate understanding of the role of lactobacilli in the vagina and the application of the knowledge gained in clinical practice for the treatment and prevention of various disorders of the normal vaginal microbiome.

The female reproductive tract microbiome and obesity

High-throughput 16S rRNA sequencing has allowed us to identify novel microorganisms and their relationships in the female reproductive tract. However, in obese patients, the female reproductive tract microbiome, unlike the intestinal microbiome, has been understudied. Here, the literature review analyzes and describes microbiome features in the external genitalia, vagina, cervical canal, uterus, and ovaries in overweight and obese nonpregnant and pregnant females. The microbiome of the lower female reproductive tract in obese patients is characterized by increased bacterial diversity, pH, decreased Lactobacillus abundance, and increased abundance of obligate anaerobes and yeasts of the genus Candida. The endometrial microbiome in overweight and obese patients has been studied only in postmenopause and is characterized by higher Proteobacteria abundance. No data on the characteristics of the ovarian microbiome in obese patients are available. The mechanisms accounting for microbiome changes in obese patients are likely to due to the ability of adipose tissue-derived leptin and estrone to inhibit production of pituitary gonadotropic hormones resulting in blocked ovulation and lowered estradiol production in patients of reproductive age. Consequently, a decline in glycogen synthesis in the vaginal epithelium, decreased Lactobacillus abundance followed by elevated vaginal mucus pH value and number of obligate anaerobes, including those associated with bacterial vaginosis are observed. Weight loss can have a beneficial effect on the state of the vaginal microbiome, restoring normal Lactobacillus abundance.

Management of a granulosa-theca cell tumor in a female Rottweiler

A 4-year, multiparous Rottweiler female dog, was presented for breeding management. Dog had clinical signs of proestrus and estrus for > 6 weeks, including cornification of vaginal epithelium and low circulating progesterone concentrations (< 2 ng/ml). Transabdominal ultrasonography of the reproductive tract and measurement of circulating hormones (antimüllerian hormone, Inhibin-B, and progesterone) suggested granulosa-theca cell tumor. Affected ovary was removed via laparoscopy. Histopathology confirmed granulosa-theca cell tumor; dog resumed cyclicity 4 months after surgery, had normal estrus and was successfully mated with a young stud dog. Dog was diagnosed pregnant (30 days after LH surge) via transabdominal ultrasonography; 4 amniotic sacs were detected, and 3 grew to term but were not viable at delivery (via cesarian surgery).

First detection and impact of bovine herpesvirus type 4 on dairy cattle reproduction in Thailand

Background and Aim: Postpartum reproductive tract infections pose significant challenges to dairy farms, leading to economic losses due to reduced fertility associated with uterine inflammation. In veterinary practice, numerous research groups have explored the underlying causes of subfertility in cows, including surveying endemic viral infections related to endometritis in local areas. This study investigated bovine herpesvirus 4 (BoHV-4) infection in Thai dairy herds and assessed its impact on endometritis and subsequent reproductive outcomes. Materials and Methods: The present study analyzed BoHV-4 DNA in various samples, including milk, blood, and endometrial tissue, from 44 Holstein-Friesian cows 21–47 days postpartum across five dairy herds in Central Thailand. BoHV-4 glycoprotein B and thymidine kinase DNA sequences were detected using the polymerase chain reaction (PCR) and nested PCR, with sequence comparisons made to GenBank data for phylogenetic analysis. The endometritis status was diagnosed through vaginal mucus examination and endometrial cytology, with reproductive performance monitored up to the subsequent calving. Results: BoHV-4 DNA was identified in blood and endometrial tissues (15.91%) but not in milk samples. Phylogenetic analysis revealed that the local BoHV-4 strains are similar to those identified in Brazil and Japan. Notably, the presence of BoHV-4 was correlated with reduced postpartum reproductive performance, particularly extending the interval from calving to the first service. Conclusion: Our findings underscore the importance of integrating BoHV-4 genomic surveys and uterine health assessments to refine reproductive management strategies within the dairy industry. Keywords: bovine herpesvirus-4, dairy cattle, endometritis, phylogenetic analysis, reproductive performance, Thailand.

Gardnerella vaginalis, Fannyhessea vaginae, and Prevotella bivia Strongly Influence Each Other's Transcriptome in Triple-Species Biofilms

Bacterial vaginosis (BV), the most common vaginal infection worldwide, is characterized by the development of a polymicrobial biofilm on the vaginal epithelium. While Gardnerella spp. have been shown to have a prominent role in BV, little is known regarding how other species can influence BV development. Thus, we aimed to study the transcriptome of Gardnerella vaginalis, Fannyhessea vaginae, and Prevotella bivia, when growing in triple-species biofilms. Single and triple-species biofilms were formed in vitro, and RNA was extracted and sent for sequencing. cDNA libraries were prepared and sequenced. Quantitative PCR analysis (qPCR) was performed on the triple-species biofilms to evaluate the biofilm composition. The qPCR results revealed that the triple-species biofilms were mainly composed by G. vaginalis and P. bivia was the species with the lowest percentage. The RNA-sequencing analysis revealed a total of 432, 126, and 39 differentially expressed genes for G. vaginalis, F. vaginae, and P. bivia, respectively, when growing together. Gene ontology enrichment of G. vaginalis downregulated genes revealed several functions associated with metabolism, indicating a low metabolic activity of G. vaginalis when growing in polymicrobial biofilms. This work highlighted that the presence of 3 different BV-associated bacteria in the biofilm influenced each other’s transcriptome and provided insight into the molecular mechanisms that enhanced the virulence potential of polymicrobial consortia. These findings will contribute to understand the development of incident BV and the interactions occurring within the biofilm.

Demonstrating the principal mechanism of action of medical devices intended for vaginal use on reconstructed human vaginal epithelium: the case of two hyaluronic acid-containing devices

Regulation (EU) 2017/745 on medical devices (MDR) has significantly modified the rules to be adopted for MD qualifications and classification. New requirements require robust evidence on mechanisms of action (MoAs) that cannot be produced by existing common EU or ISO standards. Therefore, on a “case-by-case basis,” a new evidence-based non-clinical approach to MD qualification must be defined. In this study, an in vitro experimental approach is described to assess the physicochemical and mechanical MoA of two hyaluronic acid (HA)-based medical devices: Mucogyne® Gel and Mucogyne® Ovule for vaginal use. They both act as moisturizers and lubricants as well as a healing adjuvant by promoting the continued moisture of the vulvovaginal area. The MoA of these two products has been demonstrated by using a 3D reconstructed human vaginal epithelium (HVE) model in a homeostatic physiological state and in stressed conditions. Film forming and persistency properties were assessed on intact HVE tissues by caffeine permeation assay and Lucifer Yellow (LY) localization on HVE vertical sections. Healing properties were assessed on injured HVE tissues by trans-electrical epithelial resistance (TEER) measurements associated with histo-morphological analysis (H&E), and moisturizing efficacy was evaluated on HVE tissues cultured in dry conditions by histomorphological analysis (H&E) and aquaporin 3 (AQP3) expression and localization by immunohistochemistry (IHC). Using the same “dry” HVE model, the non-pharmacological action of the two products was addressed by CD44 (hyaluronic acid receptor) expression and localization. The results suggest that in vitro evaluations can provide robust results on a human-relevant experimental model for the intended use of the products and supports clinical data with mechanistic information which may not be achieved with in vivo studies but are particularly important for product qualification. The results also underline the specific relative efficacy of the mechanisms investigated for Mucogyne® Gel and Mucogyne® Ovule in line with their different formulation types (respectively, hydrophilic and lipophilic) that influence the action of the active ingredient HA. The present in vitro non-clinical evaluation of HVE combined with clinical investigation data obtained in women explain why Mucogyne MDs provide significant benefits in various physiological or pathological situations, including vaginal dryness and healing.

Prevotella timonensis degrades the vaginal epithelial glycocalyx through high fucosidase and sialidase activities.

Bacterial vaginosis (BV) is a common vaginal infection that affects a significant proportion of women and is associated with reduced fertility and increased risk of secondary infections. Gardnerella vaginalis is the most well-known BV-associated bacterium, but Prevotella species including P. timonensis and P. bivia may also play an important role. We showed that, similar to G. vaginalis, P. timonensis adhered well to the vaginal epithelium, suggesting that both bacteria could be important in the first stage of infection. Compared to the other bacteria, P. timonensis was unique in efficiently removing the protective mucin sugars that cover the vaginal epithelium. These results underscore that vaginal bacteria play different roles in the initiation and development of BV.

The study of vaginal wall thickness in adults based on histopathological measurements

To accurately measure the vaginal mucosa thickness across different age groups using histopathologic techniques and investigate the factors that may influence the thickness changes. This study aims to provide clinicians with objective evidence of variations in vaginal mucosal thickness, facilitating personalized medical decisions for patients. A retrospective analysis was conducted on clinical data from 348 patients who underwent local vaginal wall resection at the West China Second University Hospital, Sichuan University, from January 2021 and May 2022. The thickness of vaginal mucosa, epithelium and lamina propria was measured precisely under the microscope. And the 10th, 25th, 50th, 75th, and 90th percentile values of vaginal mucosa thickness across different age groups were counted and charted a dot-line plot. The percentile values for vaginal mucosa thickness exhibited a decreasing trend with increasing age; vaginal mucosa thickness showed significant correlations with times of delivery (P = 0.031) and age (P < 0.001), both of which were negatively associated. And vaginal mucosa thickness demonstrated no significant correlation with body mass index (BMI) (P = 0.325), times of abortions (P = 0.511), times of gestation (P = 0.101), menstrual cycle (P = 0.533), or types of delivery (P = 0.056); epithelial thickness showed significant associations with age (P < 0.001) and types of delivery (P = 0.017), both of which were negative correlations. Moreover, BMI (P = 0.429), times of abortions (P = 0.764), delivery (P = 0.079), gestation (P = 0.475), and menstrual cycle (P = 0.950) were nonassociated with epithelial thickness; lamina propria thickness displayed a significant correlation only with age (P = 0.002), and there were no obvious correlations observed between lamina propria thickness and BMI (P = 0.374), times of abortion (P = 0.417), delivery (P = 0.053), gestation (P = 0.101), types of delivery (P = 0.132) and menstrual cycle (P = 0.495). Moreover, when the age segmentation was thresholded at 35 and 50 years, both epithelial thickness and vaginal mucosa thickness were significantly correlated with age (P < 0.05). Lamina propria thickness was associated with age when the age threshold was set at 35 years (P = 0.007), whereas it showed no strong link with age when the age threshold was 50 years (P = 0.072). This study has innovatively established percentile reference values for vaginal mucosa thickness based on histopathology, furnishing clinicians with objective evidence of variations in vaginal mucosal thickness to facilitate personalized medical decisions for patients. The findings demonstrated a strong link between vaginal mucosa thickness and age, with epithelium likely playing a predominant role, while the association with lamina propria appeared to be less significant. Further research involving a larger sample size is warranted to elucidate the potential relationship with the lamina propria.

Lipopolysaccharide Triggers Luminal Acidification to Promote Defense Against Bacterial Infection in Vaginal Epithelium

The vaginal epithelium plays pivotal roles in host defense against pathogen invasion, contributing to the maintenance of an acidic microenvironment within the vaginal lumen through the activity of acid-base transport proteins. However, the precise defense mechanisms employed by the vaginal epithelium following bacterial infection remain incompletely understood. This study demonstrated that the bacterial lipopolysaccharide (LPS) potentiated net proton efflux by up-regulating the expression of Na+-H+ exchanger 1 (NHE1), without affecting other acid-base transport proteins in vaginal epithelial cells. Pharmacological inhibition or genetic knockdown of Toll-like receptor-4 (TLR4) and the extracellular-signal-regulated protein kinase (ERK) signaling pathway effectively counteracted the up-regulation of NHE1 and the enhanced proton efflux triggered by LPS in vaginal epithelial cells. In vivo studies revealed that LPS administration led to luminal acidification through the up-regulation of NHE1 expression in the rat vagina. Moreover, inhibition of NHE exhibited impaired defense against acute bacterial infection in the rat vagina. These findings collectively indicated the active involvement of vaginal epithelial cells in facilitating luminal acidification during acute bacterial infection, offering potential insights into the treatment of bacterial vaginosis.

Associations among the largest follicle, preovulatory estradiol concentrations, and predominant vaginal epithelial cells at the completion of hormonal ovarian stimulation for fixed-time artificial insemination in goats.

The objective of the present study was to investigate the association among the largest follicle (LF), preovulatory estradiol (E2), and predominant vaginal epithelial cell at the completion of hormonal ovarian stimulation for fixed-time artificial insemination (FTAI) in goats. Thirty-seven crossbred Boer does received gonadotropin-releasing hormone (GnRH) and intravaginal progesterone (P4)-releasing devices (day 0). On day 5, P4 devices were removed and does received prostaglandin F2α and equine chorionic gonadotrophin. On day 7, does received GnRH, and FTAI was undertaken. On day 7, does were divided into three groups, i.e. small-sized (3-3.9 mm; n = 5), medium-sized (4-4.9 mm; n = 8), and large-sized (≥5 mm; n = 24) according to the diameter of the ovarian LF; follicular characteristics (number and diameter) were identified, and blood samples and vaginal smears were collected. The average diameters of total antral follicles and LF and the percentage of superficial cell were greatest in large-sized LF does (p < .01). The average diameters of total antral follicle (r = .68) and LF (r = .71), number of preovulatory follicle (r = .58), and plasma E2 concentrations (r = .61) were positively correlated with the percentage of superficial cells (p < .01). The likelihood of a pregnancy outcome after the FTAI increased by 13.71 times in does with a greater average diameter of antral follicle, 14.18 times with emergence of a large preovulatory follicle, and 36.83 times with a higher percentage of vaginal superficial cells (p < .01). It was concluded that there is a relationship between the cell types of the vaginal epithelium, the diameters of the largest ovarian follicles, and the concentration of E2 in goats subjected to FTAI protocols.

Multifaceted Impact of CO2 Laser Therapy on Genitourinary Syndrome of Menopause, Vulvovaginal Atrophy and Sexual Function.

Genitourinary syndrome of menopause (GSM) encompasses a range of distressing symptoms in the vulvovaginal and/or bladder-urethral regions related to menopause changes, negatively influencing woman's quality of life and sexual activity. Fractional micro-ablative CO2 laser therapy has shown the potential to reinstate the vaginal epithelium to a condition akin to the premenopausal state, thereby ameliorating the subjective symptoms associated with GSM. We conducted a prospective, pilot study in 73 sexually active postmenopausal women treated with CO2 laser for their GSM symptoms, while assessing Vaginal Health Index Score (VHIS) and sexual function through the Female Sexual Function Index (FSFI) Questionnaire. The laser treatment resulted in a decrease in VHIS and patient-reported vulvovaginal atrophy (VVA) symptoms, with a significantly lower prevalence of vaginal itching, dryness, and burning (p < 0.001), as well as dyspareunia (p = 0.002). The occurrence of urinary incontinence, urgency, and vaginal heaviness significantly reduced, with an improvement in the staging of cystocele, either to Stage 1 or complete resolution (p < 0.001). FSFI total and domain scores were significantly higher after the treatment, indicating better sexual function, with a post-treatment score median of 25 (p < 0.001). Therefore, using a three-cycle fractional CO2 laser was an effective choice for reducing urogenital discomfort related to GSM in postmenopausal women.

Tampons as a source of exposure to metal(loid)s

BackgroundBetween 52–86% of people who menstruate in the United States use tampons—cotton and/or rayon/viscose ‘plugs’—to absorb menstrual blood in the vagina. Tampons may contain metals from agricultural or manufacturing processes, which could be absorbed by the vagina’s highly absorptive tissue, resulting in systemic exposure. To our knowledge, no previous studies have measured metals in tampons. ObjectivesWe evaluated the concentrations of 16 metal(loid)s in 30 tampons from 14 tampon brands and 18 product lines and compared the concentrations by tampon characteristics. MethodsAbout 0.2 – 0.3 g from each tampon (n = 60 samples) were microwave-acid digested and analyzed by inductively coupled plasma mass spectrometry (ICP-MS) to determine concentrations of arsenic, barium, calcium, cadmium, cobalt, chromium, copper, iron, manganese, mercury, nickel, lead, selenium, strontium, vanadium, and zinc. We compared concentrations by several tampon characteristics (region of purchase, organic material, brand type) using median quantile mixed models. ResultsWe found measurable concentrations of all 16 metals assessed. We detected concentrations of several toxic metals, including elevated mean concentrations of lead (geometric mean [GM] = 120 ng/g), cadmium (GM = 6.74 ng/g), and arsenic (GM = 2.56 ng/g). Metal concentrations differed by region of tampon purchase (US versus European Union/United Kingdom), by organic versus non-organic material, and for store- versus name-brand tampons. Most metals differed by organic status; lead concentrations were higher in non-organic tampons while arsenic was higher in organic tampons. No categoriy had consistently lower concentrations of all or most metals. DiscussionTampon use is a potential source of metal exposure. We detected all 16 metals in at least one sampled tampon, including some toxic metals like lead that has no “safe” exposure level. Future research is needed to replicate our findings and determine whether metals can leach out of tampons and cross the vaginal epithelium into systemic circulation.

Molecular identification of Coxiella burnetii in raw milk samples collected from farm animals in districts Kasur and Lahore of Punjab, Pakistan

Coxiella burnetii is the worldwide zoonotic infectious agent for Q fever in humans and animals. Farm animals are the main reservoirs of C. burnetii infection, which is mainly transmitted via tick bites. In humans, oral, percutaneous, and respiratory routes are the primary sources of infection transmission. The clinical signs vary from flu-like symptoms to endocarditis for humans’ acute and chronic Q fever. While it is usually asymptomatic in livestock, abortion, stillbirth, infertility, mastitis, and endometritis are its clinical consequences. Infected farm animals shed C. burnetii in birth products, milk, feces, vaginal mucus, and urine. Milk is an important source of infection among foods of animal origin. This study aimed to determine the prevalence and molecular characterization of C. burnetii in milk samples of dairy animals from two districts in Punjab, Pakistan, as it has not been reported there so far. Using a convenience sampling approach, the current study included 304 individual milk samples from different herds of cattle, buffalo, goats, and sheep present on 39 farms in 11 villages in the districts of Kasur and Lahore. PCR targeting the IS1111 gene sequence was used for its detection. Coxiella burnetii DNA was present in 19 of the 304 (6.3%) samples. The distribution was 7.2% and 5.2% in districts Kasur and Lahore, respectively. The results showed the distribution in ruminants as 3.4% in buffalo, 5.6% in cattle, 6.7% in goats, and 10.6% in sheep. From the univariable analysis, the clinical signs of infection i.e. mastitis and abortion were analyzed for the prevalence of Coxiella burnetii. The obtained sequences were identical to the previously reported sequence of a local strain in district Lahore, Sahiwal and Attock. These findings demonstrated that the prevalence of C. burnetii in raw milk samples deserves more attention from the health care system and veterinary organizations in Kasur and Lahore of Punjab, Pakistan. Future studies should include different districts and human populations, especially professionals working with animals, to estimate the prevalence of C. burnetii.

Fungal burden, dimorphic transition and candidalysin: Role in Candida albicans-induced vaginal cell damage and mitochondrial activation in vitro.

Candida albicans (C. albicans) can behave as a commensal yeast colonizing the vaginal mucosa, and in this condition is tolerated by the epithelium. When the epithelial tolerance breaks down, due to C. albicans overgrowth and hyphae formation, the generated inflammatory response and cell damage lead to vulvovaginal candidiasis (VVC) symptoms. Here, we focused on the induction of mitochondrial reactive oxygen species (mtROS) in vaginal epithelial cells after C. albicans infection and the involvement of fungal burden, morphogenesis and candidalysin (CL) production in such induction. Bioluminescent (BLI) C. albicans, C. albicans PCA-2 and C. albicans 529L strains were employed in an in vitro infection model including reconstituted vaginal epithelium cells (RVE), produced starting from A-431 cell line. The production of mtROS was kinetically measured by using MitoSOX™ Red probe. The potency of C. albicans to induced cell damage to RVE and C. albicans proliferation have also been evaluated. C. albicans induces a rapid mtROS release from vaginal epithelial cells, in parallel with an increase of the fungal load and hyphal formation. Under the same experimental conditions, the 529L C. albicans strain, known to be defective in CL production, induced a minor mtROS release showing the key role of CL in causing epithelial mithocondrial activation. C. albicans PCA-2, unable to form hyphae, induced comparable but slower mtROS production as compared to BLI C. albicans yeasts. By reducing mtROS through a ROS scavenger, an increased fungal burden was observed during RVE infection but not in fungal cultures grown on abiotic surface. Collectively, we conclude that CL, more than fungal load and hyphae formation, seems to play a key role in the rapid activation of mtROS by epithelial cells and in the induction of cell-damage and that mtROS are key elements in the vaginal epithelial cells response to C. albicans.

Abstract PO4-27-04: GRACE-trial: A randomized active-controlled trial for vulvovaginal atrophy in breast cancer patients on endocrine therapy

Abstract Background: Breast cancer is the most common cancer type in women worldwide. Due to hormone receptor positivity in the majority of the breast cancer tumors is endocrine therapy a crucial part in the treatment landscape of breast cancer. Selective estrogen-receptor modulators or aromatase inhibitors are the used treatment options for endocrine therapy. These medicines generate a hypoestrogenic environment by reducing circulating estrogen or by altering the effect of estrogen on tissue cells by receptor blockade. As a common side effect, vulvovaginal atrophy occurs in a majority of breast cancer patients using endocrine therapy. Vulvovaginal atrophy has a significant impact on physical and psychological wellbeing due to negative influence on quality-of-life, self-esteem and sexuality. As a consequence, adherence of endocrine therapy for the standard duration of 5 to 10 years is challenging, resulting in higher rates of therapy interruption, leading to poorer prognosis with shorter distant disease free survival. The standard treatment for vulvovaginal atrophy in postmenopausal women is based on the use of local hormonal treatment. However, when a patient has a history of breast cancer, delay of treatment and undertreatment are ubiquitous. Methods &amp; design: In this first ever prospective randomized trial breast cancer patients on endocrine therapy with vulvovaginal atrophy will be treated with the available local treatment modalities with a 1:1:1:1 randomization: estrogen, dehydroepiandrosterone, moisturizers and a co-treatment of estrogen and probiotics. Patient-reported outcomes measurements will be implemented to investigate the efficacy of the implemented treatments. These outcome measures include symptom evaluation, impact on quality-of-life and sexuality. Safety of the treatments will be evaluated by assessing systemic sex hormones concentrations. Previous research indicated no increased recurrence risk, yet no randomized trials comparing the different modalities have been published. Primary objectives: Two primary objectives have been determined. Firstly, the efficacy of the different treatment modalities will be assessed. The assessment will be based on patient-reported outcome measurements (PROMs) and clinical evaluations such as vaginal pH and the vaginal maturation index, the latter a direct microscopic evaluation of the vaginal epithelium. Secondly, safety of the different groups will be assessed. Evaluation will be done be measuring sex hormone concentrations systemically, which will be a surrogate for safety evaluation. Secondary objectives: The secondary objective in this study is the identification of microbial alterations after treatment initiation. Identification of these alterations can help in further understanding of the pathophysiology of vulvovaginal atrophy and potentially create opportunities for new treatment strategies towards vulvovaginal atrophy in breast cancer patients on endocrine therapy. Registration: The current study is registered at the European Union Clinical Trials Register (EudraCT number 2021-001921-31). Trial status: Recruitment for the study was commenced in May 2022. At time of manuscript submission, patient recruitment was ongoing. Citation Format: Glenn Vergauwen, Piet Cools, Hannelore Denys, Tom Fiers, Koen van de Vijver, Liv Veldeman, Hans Verstraelen. GRACE-trial: A randomized active-controlled trial for vulvovaginal atrophy in breast cancer patients on endocrine therapy [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO4-27-04.

Estrus detection of PE goats using the ASM application and confirmed by vagina cytology

The estrus cycle is the initial stage of increasing the milk production of PE goats. Estrus detection in PE goats is generally done manually by looking for signs of sound, restlessness, and aggression. The use of voice recording applications to detect estrus has been carried out, one of which is ASM. However, to increase the accuracy of detection based on ASM audio-visual, it is necessary to confirm the cytological status of bovine vaginal cells. This study aimed to detect estrus using audio-visual and vaginal cytology of goats. Eight female PE goats were observed on small breeders for their reproductive cycle, behavior, vocalization with ASM, and cell cytology sampling through vaginal mucus. They are processing data using Spearman rank correlation (SD). The results show aggressive behavior, frequent voices, and signs of clear discharge, and the tail is often moved. ASM detected the sound of goats in estrus at 430.0 Hz – 494.3 Hz and confirmed the superficial cell status, which predominated in PE goats. It can be concluded that the audio of goats in the 472 Hz range recorded using the ASM application can be used as a reference that goats are in estrus and ready to be mated.

In-vitro antibacterial activity of Zingiber officinale (ginger) against bacteria isolated from reproductive tract of clinical endometritic Murrah buffaloes

Endometritis, an inflammation of the innermost uterine layer, is caused by a variety of bacteria, including Gram- positive and Gram-negative aerobes and anaerobes. This study aimed to identify the active compounds in Zingiber officinale (ginger) and assess its effectiveness against common bacterial pathogens found in Murrah buffaloes suffering from endometritis. The infrared spectrum of ginger powder displayed a range of wavelengths from 3361.89/cm to 664.26/cm, indicating the presence of eight functional groups. In the hydro-ethanolic ginger extract, the spectrum ranged from 3402.8/cm to 765.35/cm, revealing 12 functional groups. A total of 42 vaginal mucus samples were collected, with E. coli being the most prevalent in 23 samples, followed by S. aureus in 21 samples, Pseudomonas spp. in 12 samples, and Campylobacter spp. in 11 samples. To evaluate the in vitro antibacterial activity of ginger extracts, antimicrobial susceptibility tests were conducted at concentrations of 2.5, 5, 10, 20, 30, 40, and 50 mg/ml. Staphylococcus aureus exhibited the highest sensitivity to the ginger extract, while E. coli showed substantial resistance. In conclusion, ginger demonstrates potent antibacterial activity against multidrug-resistant uterine pathogens and could serve as an alternative therapy to mitigate the risk of drug resistance in treating uterine infections.

Demonstrates the principle of compressing thin vaginal epithelium to the underlying fascia, in order to avoid excision of vaginal tissue.

Demonstrates the principle of compressing thin vaginal epithelium to the underlying fascia, in order to avoid excision of vaginal tissue.

Expression and localization of the mechanosensitive/osmosensitive ion channel TMEM63B in the mouse urinary tract.

The epithelial cells that line the kidneys and lower urinary tract are exposed to mechanical forces including shear stress and wall tension; however, the mechanosensors that detect and respond to these stimuli remain obscure. Candidates include the OSCA/TMEM63 family of ion channels, which can function as mechanosensors and osmosensors. Using Tmem63bHA-fl/HA-fl reporter mice, we assessed the localization of HA-tagged-TMEM63B within the urinary tract by immunofluorescence coupled with confocal microscopy. In the kidneys, HA-TMEM63B was expressed by proximal tubule epithelial cells, by the intercalated cells of the collecting duct, and by the epithelial cells lining the thick ascending limb of the medulla. In the urinary tract, HA-TMEM63B was expressed by the urothelium lining the renal pelvis, ureters, bladder, and urethra. HA-TMEM63B was also expressed in closely allied organs including the epithelial cells lining the seminal vesicles, vas deferens, and lateral prostate glands of male mice and the vaginal epithelium of female mice. Our studies reveal that TMEM63B is expressed by subsets of kidney and lower urinary tract epithelial cells, which we hypothesize are sites of TMEM63B mechanosensation or osmosensation, or both.