Search Results (75)

Candida albicans is a major fungal pathogen associated with vulvovaginal candidiasis, and rapid, sensitive detection remains challenging, particularly in amplification-free formats. Here, we report NaPddCas, a microfluidic-free, droplet-based CRISPR/Cas12a detection strategy for qualitative identification of Candida albicans DNA. Unlike conventional bulk CRISPR assays, NaPddCas partitions the reaction mixture into vortex-generated polydisperse droplets, enabling spatial confinement of Cas12a activation events and effective suppression of background fluorescence. This compartmentalization substantially enhances detection sensitivity without nucleic acid amplification or microfluidic devices. Using plasmid and genomic DNA templates, NaPddCas achieved reliable detection at concentrations several orders of magnitude lower than bulk CRISPR/Cas12a reactions. The assay further demonstrated high specificity against non-target bacterial and fungal species and was successfully applied to clinical vaginal secretion samples. Importantly, NaPddCas is designed as a qualitative or semi-qualitative droplet-dependent digital detection method rather than a quantitative digital assay. Owing to its simplicity, sensitivity, and amplification-free workflow, NaPddCas represents a practical approach for laboratory-based screening of Candida albicans infections. Full article

Background/Objectives: Self-collection devices are more widely used than ever for detecting sexually transmitted infections and cervical cancer. Despite this, we still lack a clear understanding of how well these tools actually collect and release the necessary molecular samples. This study compared the in vitro uptake and release performance of commonly used self-sampling devices for total proteins, nucleic acids, and episomal human papillomavirus type 16 (HPV-16) DNA. Methods: An artificial cervicovaginal fluid composed of phosphate-buffered saline supplemented with serum and nucleic acid extracts was serially diluted 2-fold. Each dilution was applied for 5 min to the external surfaces of a vaginal veil (Vaginal Veil Collector V-Veil UP2^TM device), a flocked swab (FLOQSwabs^®), and a commercial vaginal tampon. Non-woven surgical tissue and plastic film served as controls. Total proteins and nucleic acids were quantified by spectrophotometry, and HPV-16 DNA by real-time quantitative PCR. Results: Recovery rates for proteins and nucleic acids were highest for the vaginal veil (81% and 91%), followed by the swab (66% and 70%) and non-woven tissue (44% and 47%). In contrast, the tampon and plastic film performed poorly, releasing less than 30% of proteins and negligible amounts of nucleic acids. Episomal HPV-16 DNA release was highest for the veil (89%), compared with the swab (57%), non-woven tissue (37%), tampon (4%), and plastic film (2%). Conclusions: The vaginal veil demonstrated superior uptake and release of proteins, nucleic acids, and HPV-16 DNA at physiological concentrations. Its non-absorbent structure allows high saturation with efficient release of genital components, including microbial genomes, whereas vaginal tampons retained these components, limiting analytical recovery. Full article

The vaginal microbiome plays a crucial role in maintaining host health by preserving a balanced microenvironment. Nevertheless, the definition of a “normal” vaginal microbiome remains controversial, as its composition varies depending on factors such as ethnicity and geographical origin. In most cases, members of the genus Lactobacillus predominate in healthy vaginal microbiomes, protecting against potential pathogens through specific mechanisms such as the secretion of lactic acid and bacteriocins, among others. A reduction in Lactobacillus abundance, accompanied by an increase in anaerobic organisms, predisposes the host to the development of various pathologies. Among these pathologies is infection with human papillomavirus (HPV) and the subsequent development of cervical cancer. A progressive decline in Lactobacillus has been observed as the lesion advances in different populations worldwide. In the case of the Mexican population, several Lactobacillus have been reported in healthy microbiomes: L. gasseri, L. fermentum, L. rhamnosus, L. jensenii, L. crispatus, L. delbrueckii, L. acidophilus, and L. brevis. In contrast, genera reported in dysbiosis include Sneathia, while Brevibacterium aureum and Brachybacterium conglomeratum have been associated with HPV16 infection and/or SIL. The mere presence of some bacteria is not sufficient to modulate the cellular activity of host cells; therefore, the expression, production and activity of different proteins could be affected by the vaginal microbiome. The impact of the microbiome on host cell function is the result of different metabolites produced by the bacteria, which suppress or activate different signaling and metabolic pathways. The molecular interactions between the host and microbiome, as well as their role in cervical carcinogenesis, are still unknown. In this review, we focus on the vaginal microbiome, HPV, and the impact that the interaction of the microbiome with HPV has in cervical cancer development. Full article

Group B Streptococcus (GBS) colonization during pregnancy is a major cause of neonatal infection, yet its microbial determinants remain unclear. This pilot study compared the vaginal and gut microbiota of late-pregnancy women with and without GBS colonization to explore potential microbial cues for peripartum risk stratification. Forty-three Japanese pregnant women (GBS-Negative = 34; GBS-Positive = 9) were enrolled at 35–37 weeks of gestation. Vaginal secretions and stool were analyzed by 16S rRNA (V3–V4) sequencing using QIIME 2 with SILVA annotation and community state type (CST) classification. Vaginal communities were mainly Lactobacillus-dominant. GBS-Positive women showed a non-significant tendency toward more L. iners-dominant CST III and fewer L. crispatus-dominant CST I compared with GBS-Negative women. Prevotella, Atopobium, and Gardnerella were significantly enriched in the GBS-Positive group (false discovery rate < 0.05), whereas gut microbial diversity and composition showed no significant differences between groups. Cross-site gut–vagina genus-level correlations were generally weak and non-significant. These findings suggest that, in late pregnancy, GBS colonization is linked to subtle shifts within Lactobacillus-dominant vaginal communities, with more L. iners and bacterial vaginosis-associated genera, rather than global microbiota disruption. The apparent shift from L. crispatus- to L. iners-dominant communities is hypothesis-generating and should be confirmed in larger cohorts. Full article

Candida species are common commensals within the human microbiome but can transition opportunistically to pathogenic states when host–microbe homeostasis is disrupted. Their ability to adhere to mucosa and implanted medical devices, form thick biofilms, and invade epithelial tissues makes candidiasis particularly harmful in immunocompromised and elderly populations. This review examines the reported antifungal activity of common probiotic genera such as Lactobacillus, Bacillus, Bifidobacterium, and Saccharomyces across the oral cavity, gastrointestinal tract, and vaginal tract. The probiotic mechanisms of action, such as competitive exclusion, secretion of antifungal metabolites, and immunomodulation, are explored in detail, and research methodologies are scrutinised to assess the robustness of current evidence. This review compiles evidence from a variety of studies and clinical trials showing certain probiotic strains and formulations have the ability to significantly decrease Candida colonisation and reduce candidiasis symptom prevalence. Although outcomes vary greatly between probiotic strains tested, species of Candida targeted, and specific site of infection, it is clear that selected probiotic species and their secreted substances can have prominent anti-Candida effects and promote tangible clinical improvements. Future directions for the field of probiotic study are suggested, including the roles of prebiotics, postbiotics, and synbiotic formulations to enhance probiotic efficacy against candidiasis. Full article

Depot medroxyprogesterone acetate (Depo-Provera) has been associated with an increased risk of HIV acquisition. We have previously shown that Depo-Provera administration increases immune markers for HIV preference on peripheral and cervical CD4⁺ T cells but decreases the levels of most immune mediators at vaginal and cervical mucosa. In this study, we determined the effect of cervicovaginal secretions from women before (visit 1), one month (visit 2) and three months (visit 3) after Depo-Provera treatment on HIV infectivity ex vivo. The effect of supernatants from vaginal, endocervical, and rectal swabs and from cervical cytobrush on HIV infectivity were assessed by a single-cycle infection assay using CCR5-using HIV-luciferase reporter viruses. We found that endocervical secretions from women after Depo-Provera treatment promoted HIV infectivity. When analyzing the association between endocervical mediator changes in response to Depo-Provera, available in our previous study, and the changes in HIV infectivity pre- and post-treatment, we found that changes in IL-17 and VEGF were positively associated with changes in HIV infectivity at visit 2 compared with visit 1, whereas changes in RANTES and IL-4 were negatively associated with HIV infectivity. The negative association between RANTES and HIV infectivity was also observed at visit 3 compared with visit 1. Additionally, changes in IL-1α at visit 3 were positively associated with changes in HIV infectivity compared with visit 1. These findings suggest that Depo-Provera may increase the HIV risk by shifting the mucosal milieu that promotes HIV infectivity. Full article

Background: Antimicrobial resistance (AMR) is a major public health concern. Urinary tract infections (UTIs) account for up to 85–90% of community-acquired cases. The COVID-19 pandemic disrupted healthcare access and may have influenced resistance patterns. In this context, we retrospectively evaluated the antibiotic resistance dynamics of various bacterial strains isolated between 2019 and 2023 in a hospital unit; Methods: A total of 8217 clinical specimens (urine, wound secretions, sputum, pharyngeal exudate, nasal exudate, tracheal secretions, vaginal and cervical secretions, puncture fluids, purulent secretions, blood, ear secretions, eye secretions) were processed using standard microbiological techniques. Pathogen identification and susceptibility testing were performed with the VITEK 2 Compact system, following CLSI guidelines. Results: Following the analysis of 8217 clinical samples collected over a five-year period (2019–2023), a total of 2900 microorganisms were isolated and identified. Among these, the most frequently encountered were E. coli strains, with 1204 isolates. Urine cultures represented 71.3% of all processed samples. Out of these 5860 urine cultures, 1530 (26%) were positive. The resistance of E. coli strains to ampicillin (48–55.2%), trimethoprim/sulfamethoxazole (22.9–34%), and ciprofloxacin (21.4–31.5%) remained high throughout the period. ESBL-producing strains peaked at 17.6% in 2020, with multidrug resistance rates ranging from 14% to 22.4%. Conclusions: E. coli strains displayed persistently high resistance to ampicillin, trimethoprim/sulfamethoxazole, and ciprofloxacin, with peaks in ESBL production and multidrug resistance during the COVID-19 pandemic. These trends underscore the importance of continuous surveillance and antibiotic stewardship, with direct implications for empirical UTI therapy and broader strategies to mitigate the public health impact of antimicrobial resistance. Full article

We have recently described a murine model of vaginal secretion that allows the measurement of minute changes in vaginal secretion. Using this model, we determined that female mice experience a vaginal secretory response to the scent of males, a response regulated by circadian and estrous factors since females did not respond during their sleep phase, nor when in metestrus. Female mice can distinguish the social status of a male by scent cues and show a preference for the scent of dominant males. We therefore tested whether or not vaginal responses to male scent differ by the social status of that male. Vaginal secretory responses were measured using a recently described method employing a colorimetric thread. In addition, while we have shown that the proposed female attractant α/β farnesenes evoked a strong vaginal response in female mice, a second volatile preputial gland-derived messenger, 1-hexadecanol, has also been proposed to serve as a female attractant. Here we test whether or not 1-hexadecanol similarly stimulates a vaginal secretory response. We now report that the female vaginal secretory response differs according to the social status of the male: the urine-borne scent of dominant males elicited a vaginal response, while samples from non-dominant males did not. In related odor-preference tests we confirmed that female mice spend more time investigating the urine scent of dominant males. We additionally tested whether or not a second putative female attractant 1-hexadecanol would elicit a vaginal secretory response. Like the α/β farnesenes, 1-hexadecanol is volatile, derived from preputial glands, and induces an investigatory response in females. However female mice did not experience a vaginal secretory response to the scent of 1-hexadecanol. We did confirm that females spent more time investigating hexadecanol over vehicle, indicating that there can be a disconnect between behavioral measures of interest and a vaginal preparatory response. In summary, we find that subordinate male mice do not elicit a vaginal secretory response, indicating that male social status impacts the physiological responses of females to the prospect of coitus. We additionally find that in contrast to farnesenes, the putative female attractant 1-hexadecanol does not elicit a vaginal response. These findings underscore the potential value of this murine model and indicate that even in mice, vaginal responses are under complex regulation. Full article

The uterine microbiota plays a crucial role in maintaining postpartum reproductive health in dairy cows, and its dysregulation is closely associated with uterine diseases. Vaginal discharge characteristics serve as important clinical indicators for assessing uterine status and guiding clinical decision-making. This study employed 16S rRNA gene sequencing to analyze uterine microbial diversity in cows with different discharge types. Results revealed significant microbial shifts associated with discharge severity. Notably, Caviibacter was highly enriched (up to 60.25%) in cows with mildly purulent discharge (<50%), suggesting its potential role in early-stage endometritis. In contrast, Fusobacterium and Helcococcus dominated when purulent discharge exceeded 50%, while Bacteroides, Porphyromonas, and Peptostreptococcus prevailed in cows with malodorous or discolored secretions, indicating severe inflammation. This study extends previous findings by uncovering stage-specific microbial transitions and proposing Caviibacter as a potential early biomarker of endometritis. These insights support early diagnosis and targeted interventions, contributing to improved reproductive management and sustainable dairy farming. Full article

Tissue engineering enables autologous reconstruction of human tissues, addressing limitations in tissue availability and immune compatibility. Several tissue engineering techniques, such as self-assembly, rely on or benefit from extracellular matrix (ECM) secretion by fibroblasts to produce biomimetic scaffolds. Models have been developed for use in humans, such as skin and corneas. Ascorbic acid (vitamin C, AA) is essential for collagen biosynthesis. However, AA is chemically unstable in culture, with a half-life of 24 h, requiring freshly prepared AA with each change of medium. This study aims to demonstrate the functional equivalence of 2-phospho-L-ascorbate (2PAA), a stable form of AA, for tissue reconstruction. Dermal, vaginal, and bladder stroma were reconstructed by self-assembly using tissue-specific protocols. The tissues were cultured in a medium supplemented with either freshly prepared or frozen AA, or with 2PAA. Biochemical analyses were performed on the tissues to evaluate cell density and tissue composition, including collagen secretion and deposition. Histology and quantitative polarized light microscopy were used to evaluate tissue architecture, and mechanical evaluation was performed both by tensiometry and atomic force microscopy (AFM) to evaluate its macroscopic and cell-scale mechanical properties. The tissues produced by the three ascorbate conditions had similar collagen deposition, architecture, and mechanical properties in each organ-specific stroma. Mechanical characterization revealed tissue-specific differences, with tensile modulus values ranging from 1–5 MPa and AFM-derived apparent stiffness in the 1–2 kPa range, reflecting the nonlinear and scale-dependent behavior of the engineered stroma. The results demonstrate the possibility of substituting AA with 2PAA for tissue engineering. This protocol could significantly reduce the costs associated with tissue production by reducing preparation time and use of materials. This is a crucial factor for any scale-up activity. Full article

Vulvar hygiene is an important part of general hygiene: the goals are to clear the vulvar area of microbial and cellular debris and vaginal and fecal secretions, ensure local comfort, provide natural levels of hydration, and protect the vulvar microbiota. There are few national and international guidelines on vulvar hygiene. We searched the PubMed database up until 30 November 2024, using logical combinations of the following terms: hygiene, washing, vulva, vulvar, microbiota, hydration, syndet, soap, detergent, water, and customs. The abstracts were reviewed, and potentially relevant full-text articles were retrieved and examined. The subregions of the vulva vary with regard to the presence of sweat and sebaceous glands, the keratin content, the water content, the pH, and the microbiota (notably Lactobacillus, Corynebacterium, Staphylococcus, and Prevotella). An alteration in the vulvar microbiota can cause an imbalance in the vaginal microbiota, and vice versa. Vaginal douching may have negative effects on vulvar microbiota. Hair removal might increase the risk of long-term dermatological complications. Repeated washing with water alone exposes the stratum corneum to damage, and washing with soap alters the stratum corneum proteins and lipids, increases skin water loss, and accentuates the risk of irritation. Syndet-based products have a mild detergent effect, promotion of hydration, a suitable pH for the vulvar area, and protection of the vulvar microbiota. Syndet-based products (containing a blend of surfactants, emollients, antioxidants, and buffering agents) appear to be the most appropriate for vulvar care. Full article

The vaginal microbiota, a critical determinant of women’s health, is influenced by hormonal and metabolic parameters across the lifespan. While Lactobacillus species are beneficial markers of vaginal health, microbial composition undergoes pronounced alterations after menopause. This study aimed to elucidate the associations between vaginal microbiota composition, vaginal pH, menopausal status, and metabolic parameters in Asian women. Vaginal secretion samples were collected from 40 women (20 premenopausal, 20 postmenopausal). Full-length 16S rRNA gene sequencing was used to characterize the microbiota, categorized into Community State Types (CSTs): CST-I + II (Lactobacillus crispatus/gasseri, protective), CST-III (Lactobacillus iners, neutral), and CST-IV (anaerobic bacteria, harmful). Vaginal pH and clinical data were assessed in relation to microbial profiles. CST distribution differed significantly by menopausal status and vaginal pH. Harmful-type CST-IV was more prevalent in postmenopausal women (70% vs. 40%, p < 0.05), while CST-III was dominant in premenopausal women (45% vs. 5%). CST-IV was associated with elevated pH (median 6.00, p = 0.026) and increased abundance of anaerobes including Bacteroides, Fusobacterium, Porphyromonas, Prevotella, and Streptococcus. Oral antibiotic use reduced both beneficial and harmful CSTs, shifting toward neutral CST-III (75%, p = 0.048). Use of sodium–glucose cotransporter-2 (SGLT2) inhibitors in postmenopausal women was associated with a higher prevalence of protective CST-I + II (57.14% vs. 8.33%, p < 0.05), though no significant impact on pathogen presence was observed. This study highlights the dynamic interplay between menopausal status, metabolic interventions, and vaginal microbiota composition. Findings may inform targeted strategies to maintain vaginal health in aging populations. Full article

Seroconversion is defined as a four-fold or greater rise in antibody titers. This assay is used in human prophylactic vaccine trials to confirm HSV as the cause of genital lesions and detect subclinical latent infection. We evaluated the accuracy of seroconversion in detecting infection using a guinea pig model of genital infection. Not all animals intravaginally inoculated with HSV-2 become infected, particularly if vaccinated; therefore, we need to establish criteria to determine whether an animal is infected. Our primary analysis involved considering animals to be infected if they had any of the following: (a) genital lesions; (b) HSV-2 DNA in vaginal secretions four or more weeks after HSV-2 inoculation as a marker of reactivation from latency; or (c) HSV-2 DNA in dorsal root ganglia, the site of latency. In the second analysis, we considered animals to be infected if they had positive virus cultures from vaginal swabs obtained on day two or four post HSV-2 inoculation. In the third analysis, we considered animals to be infected if they had any condition included in the first two analyses. We collected sera prior to HSV-2 inoculation and two months later and tested the first 57 animals for seroconversion using Western blotting and gG2 IgG ELISA. The results were concordant in 54 of 57 animals (95%), and when discordant, the gG2 ELISA matched infection results as defined by the primary analysis. The remaining animals were evaluated by gG2 IgG ELISA only. A total of 43 animals were inoculated with HSV-2 but not vaccinated (No vaccine group), and 224 were vaccinated with glycoprotein or mRNA vaccines prior to HSV-2 inoculation (Vaccine group). In the No vaccine group, we detected no false positives (seroconversion without infection) but 24% to 29% false negatives (no seroconversion despite infection) depending on the criteria used to define infection. In the Vaccine group, we detected 8% to 22% false positives and 31% to 37% false negatives. The accuracy of seroconversion was 74% to 79% in the No vaccine group and 71% to 76% in the Vaccine group. These results raise concerns about using seroconversion as a diagnostic test in human vaccine trials. Alternate approaches, such as subject home swabbing for HSV DNA, should be considered as a possible replacement. Full article

Tearing and salivation are wholly dependent on the activity of exocrine (lacrimal and salivary) glands, whereas vaginal moisture and secretion rely on a combination of exudation and exocrine secretion. Exocrine gland disorders impact millions, and women with Sjögren’s Syndrome often experience dry eye and mouth as well as vaginal dryness. Cannabis users’ complaints of dry eye and ‘cottonmouth’ are well-known, but some female cannabis users also report vaginal dryness. The regulation of vaginal secretion by the cannabinoid signaling system is essentially unstudied. We recently reported that despite their small size and nocturnal nature, laboratory mice have measurable basal vaginal moisture and pheromone-stimulated secretory responses that are regulated by circadian and estrous factors. We tested the regulation of vaginal moisture by cannabinoid CB1 receptors in this model. We now report that the cannabinoid receptor agonist CP55940 does not alter baseline vaginal moisture but prevents a stimulated secretory response due to a local peri-vaginal effect. Chronic intermittent CP55940 reduces basal vaginal moisture but also unmasks or induces a potentiating effect for CP55940, suggesting multiple sites of action. The acute and chronic effects likely occur via CB1 receptors. Δ⁹-tetrahydrocannabinol (THC), the chief psychoactive ingredient of cannabis, a partial agonist at CB1, has no acute or chronic effects. In summary, strong acute activation of CB1 receptors in a murine model does not reduce vaginal moisture but does prevent a pheromone-stimulated vaginal secretory response. In contrast, chronic intermittent CB1 activation reduces baseline vaginal moisture. The extent to which these findings translate to humans remains to be determined. Full article

Background: An imbalance in the vaginal microbiota, often characterized by reduced lactobacilli, paves the way forth for opportunistic bacteria from the gastrointestinal tract. The presence of aerobic bacteria in the genital tract during pregnancy can have negative outcomes on the pregnancy. Peripartum infections, when not adequately managed, can significantly impact maternal and neonatal health. Antimicrobial resistance poses an escalating global health threat, with newborns particularly vulnerable. Methods: This study constitutes a retrospective observational analysis, encompassing all microbial strains isolated from pregnant women admitted to the “Pius Brînzeu” Clinical County Emergency Hospital in Timișoara, Romania for various infectious diseases over one year. We analyzed 274 samples from 246 pregnant women, of which 242 were cervical samples, 23 urine cultures, 3 wound secretions, 3 amniotic fluids, 1 peritoneal cavity fluid, 1 sputum, and 1 hemoculture. Results: In cervical samples, Group B Streptococcus (GBS) was the most prevalent, representing 42.46% of the isolates. E. coli was the second most frequent at 30.16%, followed by K. pneumoniae at 11.9%, S. aureus at 8.73%, C. albicans at 2.78%, and other species at 3.97%. A total of 9.63% of cervical GBS isolates exhibited resistance to penicillin, while 23.36% were identified as multi-drug resistant (MDR). Methicillin-resistant S. aureus (MRSA) and MDR S. aureus strains were identified in 50% and 54.54% of the S. aureus-positive cervical samples, respectively. Conclusions: Recognizing the implications of maternal infection or colonization, especially with antimicrobial resistance bacteria, aids in assessing risks during pregnancy. Full article