Vaginitis Research Articles

B-117 Rapid Classification of Cleanliness Grades of Vaginal Discharge through Computational Analysis of SERS Spectra via Variational Auto-Encoder Model And Long Short-Term Memory Networks

Abstract Background Classification of cleanliness grades of vaginal discharge plays a pivotal role in clinical laboratories, serving as indicators for the presence of various vaginal infections such as bacterial vaginosis and candidal vulvovaginitis, etc. The conventional method for assessing vaginal discharge cleanliness involves manual microscopy, a technique susceptible to variations based on the examiner's expertise and subjective judgment, hindering the attainment of swift and accurate diagnoses. Methods This study introduces a novel approach by integrating Surface-Enhanced Raman Spectroscopy (SERS) with a deep learning algorithm for the rapid classification of vaginal discharge cleanliness grades. The proposed classification method combines the Variational Auto-Encoder (VAE) and Long Short-Term Memory (LSTM) to process SERS spectra. The quality of the training data is assessed using the signal-to-noise ratio (SNR) as a standard indicator. Results The application of the VAE-LSTM algorithm significantly enhances classification performance, providing swift and accurate results while reducing the required time. Comparative analyses with various classical machine learning algorithms and evaluation metrics reveal the superior predictive capability and time efficiency of the VAE-LSTM algorithm, demonstrating 100% accuracy and a fitting time of 2 seconds. Blind testing of SERS spectra collected from unknown patients further validates the reliability of the proposed method. Conclusions This study presents a promising and practical approach for the rapid classification of vaginal discharge cleanliness grades. The integration of the SERS technique with the VAE-LSTM algorithm not only ensures accuracy and efficiency in clinical diagnosis but also lays the foundation for advancements in non-invasive diagnostic technologies in gynecological healthcare.

Vaginal Tritrichomonas foetus infection in mice as an in vivo model for drug development against Trichomonas vaginalis.

Trichomonas vaginalis is the causative agent of the common sexually transmitted disease, trichomoniasis, which affects more than a hundred million people worldwide. Metronidazole and tinidazole, agents belonging to the 5-nitroheterocyclic class of antimicrobials, are most often used to treat infection, but increased resistance has been reported and adverse effects of these drugs can be significant. Consequently, an urgent need exists for the development of novel drug entities against trichomoniasis. Critical for antimicrobial drug development is the demonstration of in vivo efficacy. Murine models of vaginal T. vaginalis infection are unreliable for unknown reasons. Meanwhile, murine infections with the related bovine pathogen, Tritrichomonas foetus, tend to be more robust, although susceptibility to different antimicrobials might differ from T. vaginalis. Here, we explored the utility of T. foetus infection as a surrogate model for drug development against T. vaginalis. Four different T. foetus strains caused robust vaginal infection in young mice, while none of four diverse T. vaginalis strains did. Comparison of drug susceptibility profiles revealed that T. foetus and T. vaginalis were similarly susceptible to a range of 5-nitroheterocyclic and gold(I) compounds. By comparison, proteasome inhibitors were 10- to 15-fold less active against T. foetus than T. vaginalis, although one of the proteasome inhibitors, bortezomib, had low micromolar activity or better against multiple strains of both trichomonads. Different strains of T. foetus were used to demonstrate the utility of the murine vaginal infection models for in vivo efficacy testing, including for bortezomib and a gold(I) compound. The differences in susceptibility to proteasome inhibitors may be partially explained by differences in the proteasome subunit sequences between the two trichomonads, although the functional relevance of the proteasome was similar in both organisms. These findings indicate that T. foetus can serve as a reliable surrogate model for T. vaginalis in vitro and in murine infections in vivo, but caution must be exercised for specific drug classes with targets, such as the proteasome, that may display genetic divergence between the trichomonads.

Identifying changes in vaginal fluid using SERS: Advancing diagnosis of vulvovaginal candidiasis

Vaginal infections, medically termed vaginitis, encompass a spectrum of symptomatic presentations arising from disturbances within the vaginal microflora. The conventional diagnostic approach relies on microscopic examination of wet preparation of vaginal discharge, considered the ‘gold standard’ in clinical practice. Complementary to this, culture-based methodologies are often employed to reinforce diagnostic accuracy. However, challenges such as subjectivity in result interpretation, resource-intensive requirements regarding skilled personnel, and reagent utilization underscore the need for alternative diagnostic strategies.In this article, we demonstrate surface-enhanced Raman spectroscopy (SERS) and partial least squares regression (PLSR) techniques to elucidate the molecular signatures present in vaginal fluids, accounting for various influencing factors, including disruptions in the natural microflora, vaginal irrigation practices, and contraceptive usage. Furthermore, we investigated the spectral manifestations associated with vulvovaginal candidiasis (VVC) relative to control samples. Each clinical specimen underwent meticulous characterization encompassing microbial composition, pH levels, purity, and other pertinent parameters.Our findings unveil significant associations between extraneous inflammatory factors such as vaginal irrigation and diminished sample purity with alterations in SERS signals. Conversely, the day of the menstrual cycle phase exhibits negligible influence on spectral profiles. Notably, VVC samples demonstrated diverse spectral responses correlating with the abundance of pathogenic bacteria. These explorations hold promise in paving the path towards developing a novel intrinsic framework for the diagnosis of vaginitis.

Vaginal lactobacilli produce anti-inflammatory β-carboline compounds

The optimal vaginal microbiome is a Lactobacillus-dominant community. Apart from Lactobacillus iners, the presence of Lactobacillus species is associated with reduced vaginal inflammation and reduced levels of pro-inflammatory cytokines. Loss of Lactobacillus-dominance is associated with inflammatory conditions, such as bacterial vaginosis (BV). We have identified that Lactobacillus crispatus, a key vaginal bacterial species, produces a family of β-carboline compounds with anti-inflammatory activity. These compounds suppress nuclear factor κB (NF-κB) and interferon (IFN) signaling downstream of multiple pattern recognition receptors in primary human cells and significantly dampen type I IFN receptor (IFNAR) activation in monocytes. Topical application of an anti-inflammatory β-carboline compound, perlolyrine, was sufficient to significantly reduce vaginal inflammation in a mouse model of genital herpes infection. These compounds are enriched in cervicovaginal lavage (CVL) of healthy people compared with people with BV. This study identifies a family of compounds by which vaginal lactobacilli mediate host immune homeostasis and highlights a potential therapeutic avenue for vaginal inflammation.

Cross-Sectional Study on the Correlation Between Vaginal Microecology and High-Risk Human Papillomavirus Infection: Establishment of a Clinical Prediction Model.

High-risk human papillomavirus (HR-HPV) is a significant risk factor for cervical precancerous lesions and cancer. This study aimed to investigate the relationship between vaginal microecology and HR-HPV infection and to evaluate the clinical applicability of vaginal microecology in predicting HR-HPV infection. Overall, 2000 women with simultaneously detected vaginal discharge and cervical HPV were selected between March 2022 and March 2023, including 241 and 1759 cases in the HR-HPV positive and HPV negative groups, respectively. No significant differences were found in age, vulvovaginal candidiasis, trichomonas vaginitis, and β-N-acetylglucosaminosidase between the two groups (P>0.05). Significant differences were observed in Lactobacillus deficiency, bacterial vaginitis (BV), aerobic vaginitis (AV), glucuronidase (GUS), sialidase (SNA), and leukocyte esterase (LE) between the two groups (P<0.05). In the multivariate logistic regression equation, Lactobacillus deficiency, BV, AV, SNA, LE, and GUS were risk factors for HR-HPV infection (P<0.05). Three prediction models, namely, logistic regression, decision tree, and random forest, were established to rank the importance of the predictors. BV ranked first among the three prediction models. The logistic regression model demonstrated the highest accuracy in predicting the risk of HR-HPV infection. The calibration curve of the logistic regression model showed a strong correlation between the predicted and actual probabilities, and decision curve analysis revealed that the prediction model had good clinical applicability. Overall, vaginal microecology imbalance was closely associated with cervical HR-HPV infection, particularly BV and AV. The logistic regression model for the risk of HR-HPV infection based on six predictive factors (BV, AV, LE, SNA, Lactobacillus deficiency, and GUS) had good accuracy and clinical applicability.

Fetomaternal outcomes in vaginal discharge during pregnancy: A prospective comparative study

Background: Vaginal discharge (VD) is a common symptom in women during the reproductive period, especially during pregnancy. Pathological VD during pregnancy is an important public health concern because it has a sociopsychological impact on women and influences fetomaternal outcomes. Aims and Objectives: The study aims to estimate the prevalence of pathological discharge in pregnant women. The study also focuses on the demographic profile and various risk factors associated with pathological VDs and their consequences on fetal and maternal health. Materials and Methods: A prospective observational study was conducted in the Department of Obstetrics and Gynecology in a tertiary care center in Northern India. A total of 150 pregnant women with complaints of VD in the second and third trimesters with confirmation on speculum examination were recruited and evaluated for the etiology of discharge. Patients were followed until delivery and data regarding fetomaternal outcomes was collected. Results: The prevalence of pathological VD among enrolled women was 39.33% and various etiologies were candidiasis (15.33%), bacterial vaginosis (10.66%), aerobic vaginitis (8.67%), and trichomoniasis (4.67%). These infections were significantly related to pre-term delivery and pre-term pre-labor rupture of membranes. Furthermore, the incidence of prematurity, and low birth weight neonates requiring neonatal intensive care unit care was significantly high in babies born to mothers with pathological VD. Conclusion: Vaginal infections affect both maternal and neonatal health, thus, increasing the likelihood of poor perinatal outcomes. Hence, we propose that routine screening for pathological VD for all pregnant women should be introduced into standard antenatal care.

The association between lifestyle factors and the composition of the vaginal microbiota: a review.

The vaginal microbiota offers valuable insights into women's sexual health and the risk of developing sexually transmitted infections (STIs) and bacterial vaginosis. Despite the public health implications of changes in the vaginal environment, existing data on this topic remain sparse. Following the PRISMA statement guidelines, we consulted five bibliographic databases, focusing on five main daily habits and behaviors. We included only studies published up to October 2023, investigating the influence of personal hygiene, sexual behaviors, hormonal contraception, smoking, alcohol consumption, and psychosocial stress on the vaginal microbiota using next-generation sequencing. Based on our inclusion criteria, we incorporated 37 studies into this review. Hormonal contraception and personal hygiene were found to promote eubiosis of the vaginal microbiota. In contrast, sexual behaviors, smoking, alcohol consumption, and psychosocial stress were associated with an increased susceptibility to bacterial vaginosis, STIs, and severe pelvic inflammatory diseases due to a modified vaginal microbiota. Black ethnicity emerged as a confounding factor, with this population showing unstable vaginal microbiota. Oral contraception and a stable male sexual partner were found to favor Lactobacillus colonization, acting as a protective factor. Conversely, non-hormonal contraception and unprotected or non-penile/vaginal sexual activity increased the incidence of vaginal inflammation and bacterial vaginosis by disturbing the vaginal microbiota and reducing Lactobacillus abundance. Daily habits and lifestyle can influence the composition of the vaginal microbiota, thereby affecting vaginal health. Disturbances in the vaginal microbiota could be associated factors for STIs and vaginosis. Therefore, prioritizing more appropriate management of the vaginal microbiota is crucial.

Research progress of hydrogels in the prevention of pelvic inflammatory disease

AbstractPelvic inflammatory disease (PID) is a critical global health concern with the potential to lead to adverse outcomes, including infertility and chronic pelvic pain. Since PID is often caused by ascending vaginal infections or urinary tract infections, understanding the treatment of both is critical to preventing PID. Meanwhile, the emergence of drug‐resistant and persistently infected strains poses a growing challenge. This review discusses current clinical treatments for the prevention of PID from the physiologic basis of PID, as well as summarizes the advantages and research progress of hydrogels in the prevention of PID. In contrast to conventional treatments, hydrogels serve as excellent vehicles for vaginal drug delivery, maintaining the presence of the drug at the target site and controlling its release. In the context of urinary tract infections (UTIs), hydrogels are employed primarily as coatings on catheters to prevent and treat catheter‐associated UTIs. Finally, this review summarizes the limitations of hydrogels in PID prevention and future directions for development with the aim of elucidating avenues for clinical treatment of PID and informing further research.

Formulation Development of Mucoadhesive Vaginal Gel of Secnidazole and Voriconazole.

Purpose: Over the preceding decades, developing conventional drug delivery in to modified form has acquired importance in the pharmaceutical industry. Vaginal drug delivery system offers localized drug administration of drug and also avoids unwanted side effects of oral drug delivery. Vaginal drug delivery system has significant role in medical practice. Even though VDDS has a lot of advantages over the other routes of administration. The goal of developing the Mucoadhesive Vaginal Gel was to achieve the synergistic effect of voriconazole and secnidazole in the treatment of bacterial and fungal vaginal infections. Method: Mucoadhesive antimicrobial vaginal gel was optimised using Design expert software using Carbopol 934, HPMC K4M polymers. Result: Formulation was evaluated for various rheological parameters ,antimicrobial activity, invitro drug release and stability study. The spreadability and pH was found to be 5.5 to 7.4 cm and 4.12. Its compatible with vaginal pH. The selected optimized F8 formulation is the best formulation and the drug ratio is 2:1. and optimum gel viscosity is 1874 cps. antimicrobial study revealed that faster drug release of test formulation was compared to marketed product. It indicates as inhibition zone. Conclusion: The combination of drugs can serves as better option for treating vaginitis, compared to single drug formulation. Mucoadesion increases contact time of drug with affected area which rendered formulation more effective.

Abstract C017: Race-Specific Patterns of Vaginal Microbiome and HPV Infections Predict Risk of Precancerous Cervical Lesions

Abstract Background: Our previous research indicated that high-grade squamous intraepithelial lesions (HSIL), precursors to cervical cancer (CCa), are influenced by the vaginal microbiome (VMB) differentially by self-reported race. VMB enriched for certain anti-inflammatory Lactobacillus species confer protection against the risk of HSIL for white, but not Black women. Black women exhibit greater VMB diversity (associated with HPV tumorigenesis) and a lower prevalence of anti-inflammatory Lactobacillus species. But not all Black women with diverse VMB develop HSIL. Objective: Given HPV's essential role in cervical cancer, we aimed to assess the influence of sociodemographic, clinical, and microecological HPV attributes on HSIL risk within the context of the vaginal microbiome's variation by race. Methods: The VMB and HPV profiles of 1,168 women were analyzed alongside over 100 sociodemographic and clinical variables. Spearman’s correlation, Point-Biserial Correlation, chi-squared tests, and machine learning models, were employed to evaluate associations between sociodemographic, clinical, and microecological variables with HSIL, with a particular focus on the impact of race on these relationships. Results: The cohort was predominantly Black (72%), showing no significant race-based differences in HPV positivity (45%). However, HSIL incidence was nearly twice as high in Black (7%) compared to White women (4%). Women with HSIL had a higher prevalence of non-Lactobacillus-dominant VMB (68% vs. 51%) and were more likely to be HPV+ (73% vs. 44%) than those without HSIL. Co-infection with multiple HPV strains was associated with a higher risk of CIN3. Specifically, HPV16, HPV56, HPV58, HPV33, and HPV42 infections were significantly associated with higher CIN3 risks. The random forest model identified the relative abundance of Lactobacillus crispatus as the most essential factor in predicting CIN3 risk, followed by the number of HPV strains per sample. However, differential abundance analysis specifically indicated lower L. crispatus levels were associated with HSIL in white but not Black women. Conversely, HPV abundance was only associated with HSIL in Black women. Additionally, Black patients with 2 or more HPV co-infections had a higher likelihood of being diagnosed with CIN3+ compared to white counterparts. ML models highlighted different HPV-specific HSIL predictors by race: HPV number, and HPV 56, 16, and 33 in Blacks; sexual partners, HPV 90, 56, and 16 in whites. Conclusions: Our findings reveal race-specific interplays among individual risk factors, HPV infection, and VMB composition in HSIL development. The identification of key microbial and viral predictors highlights the importance of personalized approaches in preventing and managing HSIL, particularly among racially diverse populations. Citation Format: Katherine Y. Tossas, Bin Zhu, Katarzyna Tyc, Myrna Serrano, Jerome Strauss, Gregory Buck. Race-Specific Patterns of Vaginal Microbiome and HPV Infections Predict Risk of Precancerous Cervical Lesions [abstract]. In: Proceedings of the 17th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2024 Sep 21-24; Los Angeles, CA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2024;33(9 Suppl):Abstract nr C017.

The In Vivo Effect of Water-Based Lubricants on the Vaginal Microbiome of Women from Varying Age Groups: Exploratory Analysis of a Randomized Controlled Trial.

Vaginal mucosa undergoes physiological changes across the lifespan, such as increased pH and reduced natural lubrication which are known to impact vaginal commensal microorganisms, hence increasing the chances of vaginal infections. An improved understanding of vaginal microbiome composition in different age groups and the effect of social behaviors, such as the use of personal lubricants, could facilitate the development of new strategies to maintain good vaginal health. The objective of this study was to assess the effect of water-based lubricants on the human vaginal microbiome. Fifty females from three age groups (18-29, 30-44, and 45-65 years) with mild-to-moderate vaginal dryness were randomized to one of five lubricants (four of which were formulated to meet expert guidance on osmolality and pH). Subjects entered the study at tolerance or treatment phase (vaginal intercourse minimum once a week using assigned lubricant). Four vaginal swabs per participant were sampled during pre-("baseline"), post-first ("2 h", "24 h") and post-last ("4 weeks") lubricant application to assess bacterial and fungal diversity via amplicon sequencing. Vaginal pH and relative humidity were measured at baseline, 2 h, and 24 h post-lubricant application. Relative bacteriome abundance was statistically compared between timepoints for each lubricant group. Vaginal moisture, age, BMI, and pH were correlated with bacteriome relative abundance. Lactobacilli and Gardnerella sp. Were predominant across participants. Repeated lubricant application did not significantly alter the vaginal bacteriome during 4 weeks of product use (p > 0.05) when measured by relative abundance and alpha-diversity index. Bacteriome diversity and abundance differed significantly between age groups at baseline whereas lactobacilli relative abundance was negatively associated with age and vaginal pH.

Synthetic bacterial consortia transplantation attenuates vaginal inflammation and modulates the immune response in a mouse model of Gardnerella vaginalis-induced bacterial vaginosis

This study aimed to evaluate the efficacy of synthetic bacterial consortia transplantation (SBCT) and compare it with VMT (vaginal microbiota transplantation) in a mouse model of Gardnerella vaginalis-induced Bacterial vaginosis (BV). A murine model of G. vaginalis-induced BV was established, and mice were treated with SBCT, VMT, or saline. Histopathological changes, inflammatory cytokine levels, pro-inflammatory biomarker expression, helper T cell transcription factor expression, and vaginal microbiota composition were assessed. SBCT and VMT effectively suppressed G. vaginalis growth, reduced inflammation, and restored vaginal microbiota diversity. Both treatments attenuated epithelial damage, downregulated pro-inflammatory cytokines (IL-1β and IL-8), and upregulated the anti-inflammatory cytokine IL-10. SBCT and VMT also inhibited NF-κB activation, suppressed IL-17 expression, and enhanced Foxp3 expression in vaginal tissues. SBCT is a promising therapeutic approach for treating BV, as it effectively modulates the immune response and restores vaginal microbiota diversity in a mouse model of G. vaginalis-induced BV.

Progesterone promotes CXCl2-dependent vaginal neutrophil killing by activating cervical resident macrophage-neutrophil crosstalk.

Vaginal infections in women of reproductive age represent a clinical dilemma with significant socioeconomic implications. The current understanding of mucosal immunity failure during early pathogenic invasions that allows the pathogen to grow and thrive is far from complete. Neutrophils infiltrate most tissues following circadian patterns as part of normal repair, regulation of microbiota, or immune surveillance and become more numerous after infection. Neutrophils are responsible for maintaining vaginal immunity. Specific to the vagina, neutrophils continuously infiltrate at high levels, although during ovulation, they retreat to avoid sperm damage and permit reproduction. Here we show that, after ovulation, progesterone promotes resident vaginal macrophage-neutrophil crosstalk by upregulating Yolk sac early fetal organs (FOLR2+) macrophage CXCl2 expression, in a TNFA-patrolling monocyte-derived macrophage-mediated (CX3CR1hiMHCIIhi-mediated) manner, to activate the neutrophils' capacity to eliminate sex-transmitted and opportunistic microorganisms. Indeed, progesterone plays an essential role in conciliating the balance between the commensal microbiota, sperm, and the threat of pathogens because progesterone not only promotes a flurry of neutrophils but also increases neutrophilic fury to restore immunity after ovulation to thwart pathogenic invasion after intercourse. Therefore, modest progesterone dysregulations could lead to a suboptimal neutrophilic response, resulting in insufficient mucosal defense and recurrent unresolved infections.

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.

Silver secnidazole nano-hybrid emulsion-based probiotics as a novel antifungal formula against multidrug-resistant vaginal pathogens.

This study presents a novel approach to manage vaginal infections due to Candidiasis, utilizing a novel silver secnidazole nano-hybrid emulsion (Ag-Secn-NHE)-based probiotics and free Ag-Secn-NHE. Ag-Secn-NHE was prepared by simple homogenization‒ultrasonication technique and validated by using a ultraviolet‒visible scan, dynamic light scattering, transmission electron microscopy (TEM), Fourier-transform infrared spectroscopy, and zeta potential. Saccharomyces cerevisiae (RCMB 002Y001) is the most effective probiotic-producing organism that demonstrates significant effects when combined with Ag-Secn-NHE. Ag-Secn-NHE-based probiotics showed significant antifungal effect compared to free Ag-Secn-NHE, silver nitrate, silver nanoparticles, secnidazole, secnidazole nanoemulsion, and commercial vaginal wash against multidrug-resistant vaginal pathogens. The highest inhibitory effect was achieved with Ag-Secn-NHE-based probiotic against Candida auris, Candida albicans, and Cryptococcus neoformans with minimal inhibitory concentration (MIC) 0.625±0.002, 0.00625:1.25±0.012 and 0.00625:1.25±0.032mg/mL, respectively, in comparison with Ag-Secn-NHE that show MIC at 0.00625:1.25±0.612, 0.0125:2.5±0.812, and 0.0125:2.5±0.112mg/mL (Ag:Secn). Ag-Secn-NHE-based- probiotic show minimum fungicidal concentration (MFC) at range from 2.5 to 20mg/mL, wherever free Ag-Secn-NHE show MFC range from 5 to >20mg/mL. Additionally, Ag-Secn-NHE-based probiotics have 75% inhibition of biofilm formation against C. auris and 60% inhibition of biofilm formation against both Cryptococcus neoformans and C. albicans in comparison with free Ag-Secn-NHE. Time-kill curves showed that the antifungal effect of Ag-Secn-NHE-based probiotics was fungistatic at 2MIC value after 4h and after 16h for Ag-Secn-NHE. TEM photographs showed that C. auris cells treated with Ag-Secn-NHE-based probiotic formula revealed severe deformations and distored ultrastructural changes. furthermore, results indicated that the Gamma radiation up to 15kGy increases production of Ag-Secn-NHE in comparison with non-irradiated one.

Preparation and in- vitro Evaluation of Bioadhesive Vaginal Film of Tinidazole

Bioadhesive vaginal film is becoming a more effective and convenient dosage form compared to suppositories and gel for vaginal application as a method for the treatment of vaginal infections. This study aimed to prepare and characterize bioadhesive films of tinidazole to be used for vaginal delivery over a prolonged period to treat local infections. Fourteen formulas of tinidazole films were prepared by solvent evaporation method. Each film was composed of 10 mg tinidazole with different ratios and concentrations of a polymeric combination of polyvinyl alcohol/ polyvinyl pyrrolidone (PVA/PVP) or polyvinyl alcohol/ hydroxypropyl methylcellulose (PVA/HPMC) for formulas A1-A7 and B1-B7, respectively. The prepared films were evaluated for their physicochemical characteristics, content uniformity, swelling, mucoadhesive strength, mucoadhesive time, and drug release. All the prepared films were transparent, had a uniform thickness (0.1-0.15 mm), and exhibited sufficient flexibility with drug content uniformity. The mucoadhesive strength of films was in the range of 28 ± 0.2 to 75 ± 0.01 g. Films containing PVA/HPMC showed higher swelling and lower mucoadhesive strength than those containing PVA/PVP polymeric combination. The in vitro drug release study showed no significant differences (p &gt; 0.05) among films containing PVA/PVP or PVA/HPMC polymeric combinations. Formulas A3 and B4 were considered as the optimal bioadhesive films containing 3% of PVA: PVP (4:1) and 4% of PVA: HPMC (2:1), respectively as they showed complete flexibility, acceptable mucoadhesive strength and time (43 ± 0.1, 45 ± 0.23 g and 30 ± 0.02, 34 ± 0.01 min, respectively) and prolonged drug release rate. In conclusion, films of formulas A3 and B4 possess the potential as a delivery system for tinidazole for the treatment of vaginal infections at the site of application.

A Metagenomics Pipeline to Characterize Self-Collected Vaginal Microbiome Samples.

Vaginitis is a widespread issue for women worldwide, yet current diagnostic tools are lacking. Bacterial vaginosis (BV) is the most prevalent type of vaginitis, found in 10-50% of reproductive-aged women. Current diagnostic methods for BV rely on clinical criteria, microscopy, or the detection of a few microbes by qPCR. However, many vaginal infections lack a single etiological agent and are characterized by changes in the vaginal microbiome community structure (e.g., BV is defined as a loss of protective lactobacilli resulting in an overgrowth of anaerobic bacteria). Shotgun metagenomic sequencing provides a comprehensive view of all the organisms present in the vaginal microbiome (VMB), allowing for a better understanding of all potential etiologies. Here, we describe a robust VMB metagenomics sequencing test with a sensitivity of 93.1%, a specificity of 90%, a negative predictive value of 93.4%, and a positive predictive value of 89.6% certified by Clinical Laboratory Improvement Amendments (CLIA), the College of American Pathologist (CAP), and the Clinical Laboratory Evaluation Program (CLEP). We sequenced over 7000 human vaginal samples with this pipeline and described general findings and comparisons to US census data.

Meningococcal vaccine Bexsero elicits a robust cellular immune response that targets but is not consistently protective against Neisseria gonorrhoeae during murine vaginal infection.

Retrospective epidemiological studies suggest that the licensed serogroup B meningococcal vaccine 4CMenB (Bexsero) provides some protection against the closely related pathogen Neisseria gonorrhoeae in humans. This result has been replicated in murine models of gonococcal colonization, with a gonococci-reactive humoral response and more rapid clearance of vaginal infection. However, immunization with Bexsero consistently elicits a robust humoral response but does not protect all individuals, so the correlates of protection remain undefined. Herein, we exploit the fact that Bexsero promotes clearance in only a subset of immunized mice to perform a broad analysis of the adaptive response in animals that are or are not protected. We observe that Bexsero vaccination induces high levels of anti-neisserial antibodies in both serum and the vaginal lumen, and a robust cellular response highlighted by an increase in both conventional naive and memory populations as well as unconventional lymphocyte subsets. Multiplex and flow cytometry results show that Bexsero vaccination generates a robust, multi-faceted cytokine response that spans numerous T cell subsets (TH1, TH2, Treg and TH17 responses) and that non-T non-B lymphocytes play an important role in this response, as indicated by an unbiased principal component analysis. Together, this work provides the first comprehensive analysis of the robust humoral and complex cellular response to Bexsero so as to reveal the effector mechanisms that may contribute to immunity against vaginal gonococcal infection.

Nutraceuticals in Pregnancy: A Special Focus on Probiotics.

The placenta is crucial to fetal development and performs vital functions such as nutrient exchange, waste removal and hormone regulation. Abnormal placental development can lead to conditions such as fetal growth restriction, pre-eclampsia and stillbirth, affecting both immediate and long-term fetal health. Placental development is a highly complex process involving interactions between maternal and fetal components, imprinted genes, signaling pathways, mitochondria, fetal sexomes and environmental factors such as diet, supplementation and exercise. Probiotics have been shown to make a significant contribution to prenatal health, placental health and fetal development, with associations with reduced risk of preterm birth and pre-eclampsia, as well as improvements in maternal health through effects on gut microbiota, lipid metabolism, vaginal infections, gestational diabetes, allergic diseases and inflammation. This review summarizes key studies on the influence of dietary supplementation on placental development, with a focus on the role of probiotics in prenatal health and fetal development.

Rational selection of TbpB variants elucidates a bivalent vaccine formulation with broad spectrum coverage against Neisseria gonorrhoeae.

Neisseria gonorrhoeae is the causative agent of gonorrhea, an on-going public health problem due in part to the lack of success with efforts to develop an efficacious vaccine to prevent this sexually transmitted infection. An attractive candidate vaccine antigen because of its essential function and surface exposure, the gonococcal transferrin binding protein B (TbpB) exhibits high levels of antigenic variability which poses a significant obstacle in evoking a broadly protective vaccine composition. Here, we utilize phylogenetic information to rationally select TbpB variants for inclusion into a potential gonococcal vaccine and identify two TbpB variants that when formulated together elicit a highly cross-reactive antibody response in both rabbits and mice against a diverse panel of TbpB variants and clinically relevant gonococcal strains. Further, this formulation performed well in experimental proxies of real-world usage, including eliciting bactericidal activity against 8 diverse gonococcal strains and decreasing the median duration of colonization after vaginal infection in female mice by two heterologous strains of N. gonorrhoeae . Together, these data support the use of a combination of TbpB variants for a broadly protective gonococcal vaccine.