Vaginal Epithelium Research Articles

Improvement in identification of pro-estrous mice by using a novel method of detecting vaginal mucous cells.

Accurate identification of the murine estrous cycle using vaginal exfoliative cytology is the initial and crucial step for controlled reproduction of this species. However, it is generally difficult to discriminate each stage of the cycle, and thus to select pro-estrous mice for mating. To increase the accuracy of identification of the pro-estrous stage, we re-evaluated the vaginal fold histology and modified the method of exfoliative cytology. Tissue fixation using methanol in Carnoy's solution but not paraformaldehyde, combined with Alcian blue staining but not the conventional Giemsa staining, resulted in better manifestation of mucosal cell layers in the vaginal epithelium just above the keratinized layer. This mucous layer in the fold histology was found to form specifically in the pro-estrous and late di-estrous stages, and the mucous cells exfoliated in smear samples only in the pro-estrous stage. This novel method was found, by a blinded test, to increase the rate of accurate identification of the pro-estrous stage compared to the conventional method (80% vs 50%). Consistent with this finding, the mating experiment with "pro-estrous" females selected by the novel method revealed a significantly higher success rate than that with the conventional method (78.0% vs 47.5%). Thus, our study demonstrates vaginal exfoliative mucous cells as a better potential marker to detect the "receptive" state of female mice that leads to an improved success rate of mating.

The Aqueous Extract of Dacryodes edulis (Burseraceae) Leaves Inhibits Cell Proliferation Induced by Estradiol on the Uterus and Vagina of Ovariectomized Female Wistar Rats.

Proliferation is a cellular process strongly linked to the genesis of cancer. Natural substances with antiproliferative activities are currently potential alternatives in the treatment of cancers. Dacryodes edulis, for instance, is a medicinal plant traditionally used in the treatment of cancer. Scientific studies have reported the antioxidant activity of this plant. In addition, the presence of prostate cancer chemopreventive polyphenols was reported in D. edulis extracts. Therefore, this study was aimed to evaluate the effects of the aqueous extract of D. edulis leaves on cell proliferation induced by estradiol in ovariectomized female Wistar rats. In this regard, ovariectomized (OVX) rats were cotreated with estradiol valerate (E2V) (0.75 mg/kg) and the aqueous extract of D. edulis leaves. Control groups received either the vehicle (sham-operated animals and the OVX control), E2V (0.75 mg/kg) only, or E2V (0.75 mg/kg) and tamoxifen (10 mg/kg). Treatments were administered orally for 3 consecutive days, and animals were sacrificed thereafter. Epithelial heights of the uterus and vagina were assessed. Uterine levels of total cholesterol and estradiol were determined as well. Results showed that the aqueous extract of D. edulis leaves reversed the effects of estradiol as it reduced uterine weight (p < 0.05), uterine (p < 0.05), and vaginal (p < 0.001) epithelium heights. This antiproliferative effect of D. edulis was associated with reduced tissue (uterine) levels of estradiol (p < 0.001). These results suggest that the aqueous extract of D. edulis leaves could be a potential alternative treatment for proliferation-related diseases.

Development and evaluation of mucoadhesive bigel containing tenofovir and maraviroc for HIV prophylaxis

BackgroundSexual transmission of HIV is the most common means of acquiring the disease. Topical microbicides have been investigated to prevent transmission. This study will use a specific entry inhibitor, maraviroc, and a nucleotide reverse transcriptase inhibitor (NRTI), tenofovir, a dual combination which will provide a synergist effect that can enhance the efficacy of HIV microbicides via a mucoadhesive dual compartment bigel. Bigel formulation via hydrogel organogel linkages were developed and evaluated for their physicochemical characteristics, safety, and anti-HIV efficacy. In vitro diffusion studies were performed with Franz diffusion cells having effective diffusion surface area of 1.76cm2 and receiver chamber volume of 15mL.ResultThe bigel formulations showed a viscosity ranging from 14179 to 14560 cPs and had a good spreadability and acidic pH in the range of 4.0 ± 0.34 to 5.2 ± 0.18. The bigel formulations showed good anti-HIV activity at a concentration of 0.1 μg/mL. The in vitro release study of maraviroc from the bigel formulations showed a release rate ranging from 2.675 to 3.838 μg/cm2/min½ while the release rate for tenofovir ranged from 3.475 to 3.825 μg/cm2/min½. The bigel formulations were non-toxic to the human vagina as there was < 1 log10 change in Lactobacilli crispatus viability.ConclusionThis study successfully developed a dual compartment bigel containing maraviroc and tenofovir. BG C was found to be stable and safe towards vaginal and rectal epithelium, and it actively prevented HIV transmission. This bigel has the potential for long-term pre-exposure prophylaxis prevention of HIV transmission.

The Cervicovaginal Mucus Barrier.

Preterm births are a global health priority that affects 15 million babies every year worldwide. There are no effective prognostic and therapeutic strategies relating to preterm delivery, but uterine infections appear to be a major cause. The vaginal epithelium is covered by the cervicovaginal mucus, which is essential to health because of its direct involvement in reproduction and functions as a selective barrier by sheltering the beneficial lactobacilli while helping to clear pathogens. During pregnancy, the cervical canal is sealed with a cervical mucus plug that prevents the vaginal flora from ascending toward the uterine compartment, which protects the fetus from pathogens. Abnormalities of the cervical mucus plug and bacterial vaginosis are associated with a higher risk of preterm delivery. This review addresses the current understanding of the cervicovaginal mucus and the cervical mucus plug and their interactions with the microbial communities in both the physiological state and bacterial vaginosis, with a focus on gel-forming mucins. We also review the current state of knowledge of gel-forming mucins contained in mouse cervicovaginal mucus and the mouse models used to study bacterial vaginosis.

Cytology of smears of prints with symptomatic forms of infertility in cows

Many works of domestic and foreign scientists are devoted to the study of etiological factors, pathogenesis, as well as the development of effective methods of diagnosis and therapy for symptomatic infertility of cows, however, insufficient attention is paid to cytological examination of the genital organs.So the goal of our work was to study the cytology of vaginal mucus at the symptomatic form of infertility of cows. The article presents the cytological composition of vaginal mucus from the mucous membrane of the caudal section of the cervix, the lateral surface of the vagina and the dorsal wall of the vaginal vestibule. Research was carried out with 5 groups of cows 4–5 years old cows 6 animals in each, weighing 500–600 kg, with an average productivity of 7500 kg of milk per lactation. The tests were cows were on the 18–24 day after insemination and during the sexual cycle braking, research of the joint course of uterus and hypotension of the ovarian, the presence of persistent corpus luteum and subclinical chronic endometritis, cysts and persistent corpus luteum of the ovarian body. For sub-clinical chronic endometritis, basal cells (32–41 %) are found in wet prep with vaginal mucosa, which are arranged in layers, parabasal (22–6 %) and superficial (5–21 %), but more interstitial (37–66 %), and bare neutrophil nuclei. According to hypofunction of ovaries in vaginal smears, neutrophils, vacuoles in interstitial cell nuclei (44–56 %), basophilic cells (30–43 %), superficial cells (20–21 %), parabasal cells (3–6 %). Basal cell hypofusions were (3–43%), parabasal (3–6 %), interstitial (44–56 %), superficial (20–21 %), and bilateral (12–26 %) (4–5 %) (36–55 %) and (30–33 %). It should be noted that for the functional state of the body, the cells from the cervix are flat. In addition, the drug contains single white blood cells with light cytoplasm without signs of phagocytosis. They are structurally similar to cells of vaginal smear. During the esrtus, the smear is dominated by interstitial and surface cells and white cells, and in the follicular proliferative phase, the number of surface cells increases, while the ovulator phase is characterized by the pronounced maturation of the cells, they are flat and spaced apart.

Assessing the Genomic Variability of Gardnerella vaginalis through Comparative Genomic Analyses: Evolutionary and Ecological Implications.

Gardnerella vaginalis is described as a common anaerobic vaginal bacterium whose presence may correlate with vaginal dysbiotic conditions. In the current study, we performed phylogenomic analyses of 72 G. vaginalis genome sequences, revealing noteworthy genome differences underlying a polyphyletic organization of this taxon. Particularly, the genomic survey revealed that this species may actually include nine distinct genotypes (GGtype1 to GGtype9). Furthermore, the observed link between sialidase and phylogenomic grouping provided clues of a connection between virulence potential and the evolutionary history of this microbial taxon. Specifically, based on the outcomes of these in silico analyses, GGtype3, GGtype7, GGtype8, and GGtype9 appear to have virulence potential since they exhibited the sialidase gene in their genomes. Notably, the analysis of 34 publicly available vaginal metagenomic samples allowed us to trace the distribution of the nine G. vaginalis genotypes identified in this study among the human population, highlighting how differences in genetic makeup could be related to specific ecological properties. Furthermore, comparative genomic analyses provided details about the G. vaginalis pan- and core genome contents, including putative genetic elements involved in the adaptation to the ecological niche as well as many putative virulence factors. Among these putative virulence factors, particularly noteworthy genes identified were the gene encoding cholesterol-dependent cytolysin (CDC) toxin vaginolysin and genes related to microbial biofilm formation, iron uptake, adhesion to the vaginal epithelium, as well as macrolide antibiotic resistance.IMPORTANCE The identification of nine different genotypes among members of G. vaginalis allowed us to distinguish an uneven distribution of virulence-associated genetic traits within this taxon and thus suggest the potential occurrence of putative pathogen and commensal G. vaginalis strains. These findings, coupled with metagenomics microbial profiling of human vaginal microbiota, permitted us to get insights into the distribution of the genotypes among the human population, highlighting the presence of different structural communities in terms of G. vaginalis genotypes.

The Association between Testosterone Therapy and Postoperative Complications Among Transmen Undergoing Gender Affirming Hysterectomy

Study Objective While there is increasing evidence on the safety of testosterone for transmen, little is known regarding its perioperative implications. This study seeks to investigate the association between testosterone and postoperative complications among transmen undergoing gender affirming hysterectomy. Design Retrospective cohort study of all gender affirming hysterectomies performed at a single teaching hospital in the United States over 26 months, with 6 weeks follow up to identify postoperative complications. Setting Standard laparoscopic operating room setup with patients in dorsal lithotomy. Patients or Participants 34 transmen on testosterone for at least 12 months before hysterectomy and continued at time of surgery. Interventions All patients underwent a total laparoscopic hysterectomy with bilateral salpingectomy Measurements and Main Results Data was collected through chart review of preoperative visits for covariates and testosterone dose and postoperative visits for complications. There were 8 postoperative complications including 6 transmen who experienced vaginal bleeding requiring application of hemostasis agents to the vaginal cuff. Mean weekly testosterone dose was 69.3mg among transmen who experienced a complication and 62.5mg among those who did not. Both groups had similar mean age (31.8 vs. 29.2), mean BMI (29.8 vs. 26.9), race (87.5% vs. 57.7% White, 12.5% vs. 26.9% Black), smoking status (12.5% vs. 19.2% current smokers), and estimated blood loss (55.7ml vs. 62.9ml). Univariable and multivariable logistic regressions were performed, adjusting for age, race, BMI, and estimated blood loss. The odds of experiencing a postoperative complication was 3% higher for every additional milligram of testosterone (OR 1.03, 95%CI 0.98-1.08). Conclusion Transmen on testosterone experience vaginal bleeding post-hysterectomy at a higher rate than reported in the literature for cis-women. While testosterone de-estrogenizes the vaginal epithelium, testosterone dose is not statistically significantly associated with postoperative complications including vaginal bleeding. Additional studies with larger samples and studies evaluating the effect of postoperative vaginal estrogen on bleeding among transmen are needed to better characterize this postoperative complication.

Mayer-Rokitansky-Küster-Hauser syndrome with rare findings of inferior crossed-fused renal ectopia and Gartner's duct cyst: a video case report

To describe the treatments of a patient using the laparoscopic Davydov's method for Mayer-Rokitansky-Küster-Hauser (MRKH) syndrome and ureteral reimplantation for hydronephrosis and hydroureter. Surgical video article. A consent form from the patient was obtained as appropriate; the nature of the study did not necessitate ethics committee approval. There were no conflicts of interest. University hospital. A 28-year-old woman who presented at our gynecology department with the symptoms of primary amenorrhea and difficult intercourse. She had repaired congenital rectovestibular fistula and imperforate anus at the age of 8. At physical examination, she had a phenotypically normal vulva with a vaginal small pouch (0.5 cm). Magnetic resonance imaging of the pelvis revealed normal ovaries, a primordial uterus, absence of vaginal canal, and a 4.0 × 4.2 × 4.0 cm cystic structure posterior to the bladder. Magnetic resonance urography showed right to left renal crossed-ectopia with inferior fusion, and hydronephrosis and hydroureter from the superior kidney with Grade Ⅳ vesicoureteral reflux. Karyotype was 46, XX. Saline solution 300 mL was injected into the rectovesical space with an infusion of diluted adrenaline (1:200,000). The goal of this injection was to aid in the identification of tissue planes and reduce blood loss. The space between urethra/bladder and rectum progressively was dissected. Blunt dissection was performed initially with digital separation of tissues. Then, an 8-cm-long neovaginal vault of about 3 cm in diameter was created. The mobilized peritoneum was pulled downward with eight Vicryl sutures and connected to the vaginal epithelium. By cystoscope, we found the left orifice but could not find the right orifice of the hydroureter. Then we ligated the hydroureter by 2-0 absorbable suture near the cyst and cut off the hydroureter, and then incised of all the layers at the top of the bladder to make a bladder flap. We placed the 5 Fr double J stent in the hydroureter and the bladder and anastomozed with the ureteral stump (3-0 Vicryl). Then we removed the cyst laparoscopically. We performed a purse-string stitch to create the apex of the neovagina by taking posterior serosa of the bladder, the pelvic peritoneum between the ovary and rectum, primordial uterus, and anterior rectal serosa. Measurement of the final canal length, sexual function (Female Sexual Function Index), and degree of hydronephrosis. Three days later, we started to change the vaginal mold and the patient was advised to wear it day and night for the first postoperative month. The vaginal mold had to be worn each night until normal sexual intercourse was possible. Findings confirmed the cyst was Gartner's duct cyst. One year after the surgery, the final canal length was 9 cm and Female Sexual Function Index score was 28. The ultrasound showed that the degree of hydronephrosis of upper moiety was mild. The distal Wolffian ducts in the female are absorbed but may persist as vestigial remnants (Gartner's duct cysts). A few cases of the combined urogenital-Wolffian anomalies are reported; most of them are associated with the anomalies of müllerian duct fusion, such as Herlyn-Werner-Wunderlich syndrome (uterus didelphys, obstructed hemivagina, and mesonephric duct anomalies). The embryogenesis of the combined anomalies is not completely understood. With comprehensive preoperative assessments, laparoscopic surgery could be a safe and effective treatment to these cases.

Current treatment options for genitourinary syndrome

At the onset of menopause, the urogenital symptoms begin to develop simultaneously with vasomotor symptoms: vaginal soreness, itching and dryness, dyspareunia, etc. The development of atrophic changes in the urogenital tract greatly reduces the quality of life and is a risk factor for the development of recurrent urinary tract infections. More than 60% of postmenopausal women, who do not take systemic menopausal hormone therapy (MHT), suffer from vaginal atrophy experience symptoms. The article discusses and evaluates effectiveness of various therapies for genitourinary syndrome of menopause (GSM). Nonhormonal lubricating gels are recommended as the first-line therapy. Local intravaginal administration of estrogens is used in moderate to severe vulvovaginal atrophy and in the absence of contraindications. Women with estrogen-dependent cancers can use low doses of local estrogen against the background of Tamoxifen therapy. New promising therapies for GSM are presented: ospemifene, an oral active selective estrogen receptor modulator (SERM), laser therapy, dehydroepiandrosterone. Even though the urogenital atrophy is a common disease, administration frequency of local estrogens as therapy is low. Basic principles for treatment of urogenital atrophy include alleviation of symptoms and restoration of normal blood supply to the vaginal epithelium and urothelium wall and mucous membrane. The therapy should be initiated early and the response time to therapy will depend on the degree of baseline atrophy. Vaginal moisturizers and lubricants can be used in combination with or separately from natural estrogens in cases when patients have medical contraindications to estrogen treatment.

Flt3-L enhances trans-epithelial migration and antigen presentation of dendritic cells adoptively transferred to genital mucosa

Dendritic cells (DCs) play a critical role in shaping adaptive immunity. Systemic transfer of DCs by intravenous injection has been widely investigated to inform the development of immunogenic DCs for use as cellular therapies. Adoptive transfer of DCs to mucosal sites has been limited but serves as a valuable tool to understand the role of the microenvironment on mucosal DC activation, maturation and antigen presentation. Here, we show that chitosan facilitates transmigration of DCs across the vaginal epithelium in the mouse female reproductive tract (FRT). In addition, ex vivo programming of DCs with fms-related tyrosine kinase 3 ligand (Flt3-L) was found to enhance translocation of intravaginally administered DCs to draining lymph nodes (dLNs) and stimulate in vivo proliferation of both antigen-specific CD4+ and CD8+ T cells (cross-presentation). Mucosal priming with chitosan and DC programming may hold great promise to enhance efficacy of DC-based vaccination to the female genital mucosa.

Aborte und Totgeburten bei Ziegen unter besonderer Berücksichtigung von Coxiella burnetii

Coxiellosis, caused by the bacterium Coxiella burnetii, is a reportable disease in animals and humans in Switzerland. The number of cases in farm animals and humans has risen continuously in recent years. The aim of this work was to investigate abortions and stillbirths in goats with a focus on C. burnetii, to identify excretory routes which pose a zoonotic risk and the excretion time after an acute infection. Besides the submitted fetuses, does were screened with a serological antibody test. In addition, excretion via milk, faeces and vaginal mucus were investigated in dams with fetuses tested positive for C. burnetii at 14-day intervals.C. burnetii were isolated in 8 cases (3× in the placenta, 2× in the abomasum, 3× in the placenta and abomasum) of 13 examined stillbirths/abortions. Ten abomasums of goat kids and 8 placentas were examined using modified Ziehl-Neelsen staining (ZN) according to Stamp simultaneously with a real-time PCR. Four of 18 samples were false negative using modified ZN staining according to Stamp in contrast to real-time PCR. Seven does had serum antibodies against Coxiella. The excretion of C. burnetii persisted for 63 days in the milk, for 96 days in the vaginal mucus and for 96 respectively 114 days in two does monitored extensively. Intermittent excretion could also be observed in the milk during these 63 days. The present study showed that confirmation of disease, respectively transmission cannot be based on a single test. Only combined serological antibody test and real-time PCR examinations of birth material, milk, feces and vaginal mucus can result in a conclusive diagnosis. In addition, the examination using modified ZN staining according to Stamp is less sensitive and specific than the real-time PCR examination.

VP45.28: Effects of coronavirus disease 2019 (COVID‐19) on maternal, perinatal and neonatal outcomes: a systematic review

To perform a systematic review of available published literature on pregnancies affected by COVID-19 to evaluate the effects of COVID-19 on maternal, perinatal and neonatal outcomes. We performed a systematic review to evaluate the effects of COVID-19 on pregnancy, perinatal and neonatal outcomes. We conducted a comprehensive literature search using PubMed, EMBASE, Cochrane library, China National Knowledge Infrastructure Database and Wan Fang Data until 20 April 2020 (studies were identified through PubMed alert after 20 April 2020). Eligibility criteria included COVID-19 diagnosis, patient was pregnant on admission, availability of clinical characteristics, including maternal, perinatal or neonatal outcomes. 19 studies, including 266 pregnant women with COVID-19, were included. Among the seven case-series, the maternal age ranged from 20 to 41 years and the gestational age (GA) on admission ranged from 5 to 41 weeks. The most common symptoms were fever, cough, dyspnea and fatigue. The rate of severe pneumonia was relatively low, with the majority of the cases requiring ICU admission. Almost all cases from the case-series had positive CT chest findings. There were 6 and 22 cases that had nucleic-acid testing in vaginal mucus and breast milk, respectively, which were negative for SARS-CoV2. 177 cases had delivered, of which the majority by Caesarean section. The GA at delivery ranged from 28 to 41 weeks. Over one-third of neonates were transferred to NICU. There was one case each of neonatal asphyxia and death. There were 113 neonates that had nucleic-acid testing in throat swab, which was negative for SARS-CoV2. From the case reports, there were two maternal deaths reported. The clinical characteristics of pregnant women with COVID-19 are similar to those of nonpregnant adults with COVID-19. The subject of vertical transmission of SARS-CoV2 remains controversial and more data is needed to investigate this possibility.

Surgical Outcomes of Transvaginal Neobladder-Vaginal Fistula Repair After Radical Cystectomy with Ileal Orthotopic Neobladder: A Case-Control Study.

PurposeTo present surgical methods and outcomes in women with bladder cancer (BCa) requiring correction of neobladder-vaginal fistula (NVF) after radical cystectomy (RC) with ileal orthotopic neobladder (IONB).Materials and MethodsThe medical records of 163 women who underwent RC with IONB for BCa between January 2010 and December 2018 were retrospectively reviewed. The presence of NVF was confirmed by cystoscopy and/or voiding cystography. NVF repair was performed using a transvaginal approach, which included circumferential incision of the fistula tract, creation of a plane between the neobladder serosa and the vaginal epithelium, and multi-layered transvaginal closure.ResultsDuring a median follow-up of 47.9 months, NVF was identified in 12 (8.8%) of the 163 included women. Eight (66.7%) fistulas were located in the proximal anterior vaginal wall and four (33.3%) in the vaginal apex. Median time from RC to NVF repair was 3.4 months (range, 2.1–5.6 months), median NVF size was 6.0 mm (range, 4.0–22.0 mm), and median duration of urethral Foley catheter indwelling was 24.0 days (range, 15.0–43.0 days). Initial repair of NVF was successful in ten (83.3%) patients. Two (16.7%) patients who relapsed retained IONB through the subsequent operation. Two (16.7%) patients developed severe urinary incontinence after NVF repair, requiring anti-incontinence surgery with a synthetic transobturator mid-urethral sling.ConclusionThe transvaginal approach for NVF repair is feasible, yielding successful surgical outcomes. However, women should be counseled about the risks of relapse and urinary incontinence.

Laboratorial Aspects of Cytolytic Vaginosis and Vulvovaginal Candidiasis as a Key for Accurate Diagnosis: A Pilot Study

To identify clinical, microscopic, and biochemical characteristics that differentiate cytolytic vaginosis (CV) from vulvovaginal candidiasis (VVC). The present cross-sectional study analyzed the vaginal contents of 24 non-pregnant women aged 18 to 42 years who were attended at the Genital Infections Clinic at Centro de Atenção Integral à Saúde da Mulher da Universidade Estadual de Campinas (CAISM-UNICAMP). They were diagnosed either with (CV = 8, VVC = 8) or without vulvovaginitis or vaginal dysbiosis (controls). The socio-demographic, clinical, and gynecological data were obtained from a detailed patient interview. Samples of the vaginal contents were collected for analysis of vaginal pH, gram stain, and specific fungal culture. The Kruskal-Wallis and Fisher exact tests were used to compare the differences between the groups. Odds ratios were used to compare the categorical variables. The significance level was considered at p < 0.05. Both women with CV and VVC had a lumpy vaginal discharge (p = 0,002) and vaginal hyperemia (p = 0.001), compared with controls. The inflammatory process was more intense in the VVC group (p = 0.001). In the CV group, there was statistical significance for the lactobacillus amount (p = 0.006), vaginal epithelium lysis (p = 0.001), and vaginal pH (p = 0.0002). Cytolytic vaginosis and VVC diagnoses rarely differ on clinical characteristics but have different laboratorial findings. The present study highlights the importance of conducting an accurate investigation through laboratory tests rather than clinical criteria to avoid misdiagnosis.

Healing effects of a protein scaffold loaded with adipose-derived mesenchymal stem cells on radiation-induced vaginal injury in rats.

ObjectivesCervical cancer, the most common female cancer after breast cancer, is typically treated using radiotherapy. However, pelvic radiotherapy can cause irreversible damage to the vagina, seriously affecting patients’ quality of life. In this study, protein scaffolds loaded with rat adipose-derived mesenchymal stem cells (ADSCs) were implanted into irradiated tissue to assess their healing potential.MethodsWe established a rat model of radiation-induced vaginal injury. Complexes (consisting of protein scaffolds loaded with ADSCs) were implanted into injury sites. Histological analysis were used to assess regeneration of the vaginal epithelium. RNA sequencing was used to study the therapeutic mechanism of the complexes.ResultsThe complexes promoted vaginal epithelial cell regeneration, vaginal tissue repair and improved vaginal stenosis and contracture. Compared with rats transplanted with ADSCs, rats transplanted with complexes achieved better therapeutic effects.ConclusionsProtein scaffold-ADSC complexes had a beneficial therapeutic effect on radiation-induced vaginal injury in rats and may serve as the basis of a novel therapeutic approach for radiation dermatitis.

Evaluation of different culture media to support in vitro growth and biofilm formation of bacterial vaginosis-associated anaerobes.

BackgroundBacterial vaginosis (BV) is one of the most common vaginal infections worldwide. It is associated with the presence of a dense polymicrobial biofilm on the vaginal epithelium, formed mainly by Gardnerella species. The biofilm also contains other anaerobic species, but little is known about their role in BV development.AimTo evaluate the influence of different culture media on the planktonic and biofilm growth of six cultivable anaerobes frequently associated with BV, namely Gardnerella sp., Atopobium vaginae, Lactobacillus iners, Mobiluncus curtisii, Peptostreptococcus anaerobius and Prevotella bivia.MethodsA total of nine different culture media compositions, including commercially available and chemically defined media simulating genital tract secretions, were tested in this study. Planktonic cultures and biofilms were grown under anaerobic conditions (10% carbon dioxide, 10% helium and 80% nitrogen). Planktonic growth was assessed by optical density measurements, and biofilm formation was quantified by crystal violet staining.ResultsSignificant planktonic growth was observed for Gardnerella sp., A. vaginae and L. iners in New York City III broth, with or without ascorbic acid supplementation. Biofilm quantification showed high in vitro biofilm growth for Gardnerella sp., P. anaerobius and P. bivia in almost all culture media excluding Brucella broth. Contrary, only New York City III broth was able to promote biofilm formation for A. vaginae, L. iners and M. curtisii.ConclusionsOur data demonstrate that New York City III broth relative to the other tested media is the most conducive for future studies addressing polymicrobial biofilms development as this culture medium allowed the formation of significant levels of single-species biofilms.

Morphophysiological variations of the female reproductive organs of the vespertilionid bat Myotis nigricans during its different reproductive phases

Myotis nigricans is an important species of insectivorous bat from the Vespertilionidae family, which is endemic of the Neotropics. Despite its wide distribution, no detailed study of the reproduction is found for this species. Thus, the aim of the present study was to evaluate and characterize morphological and morphometric variations of the female reproductive organs of M. nigricans during its different reproductive phases. Twenty adult females were collected, divided into four (4) sample groups, according to the reproductive status (nonreproductive, initial and advanced pregnancy and lactating), and submitted to anatomical, histological and morphometric analyses. The results demonstrated that the female reproductive organs of M. nigricans were composed of the ovaries, the uterus, the uterine tubes and the vagina, in addition to the mammary glands. The ovaries present a great number of interstitial glands and a small but persistent corpus luteum. Ovulation is simple, unilateral and non-preferential, possibly alternated in successive ovulations and may be sequential with the occurrence of postpartum estrus. The uterus is bicornuate, implantation is fundic and the placenta is chorioallantoic, discoidal and hemochorial. The vaginal epithelium is weakly keratinized and shows great disorganization during the periods of pregnancy and lactation.

Influence of preoperative preparation on the vaginal architectonics of postmenopausal women with severe genital prolapse

Patients with severe postmenopausal genital prolapse (GP) are the most clinically difficult group of women, in which surgical treatment with mesh implants presents significant difficulties due to the duration of lesions amid pronounced structural changes in the tissues of the urogenital tract under conditions of hormone deficiency. That is why it is of particular importance to study the histological characteristics of the vaginal mucosa in women with GP, who must conduct surgical treatment of vaginal access.&#x0D; Aim. To study the histological features of the vaginal mucosa in postmenopausal women with severe GP in preparation for surgical treatment with vaginal access.&#x0D; Materials and methods. The study involved 70 postmenopausal women suffering from GP of stage III, IV in combination with genitourinary menopausal syndrome. Patients were randomized blindly into two groups. The therapy was carried out using vaginal forms of estrogen and progesterone preparations in combination with lactobacilli. In the comparison group, preoperative preparation was not performed. All patients underwent clinical and laboratory examination: general examination and gynecological examination, morphological examination of biopsy samples of the mucous membrane of the anterior vaginal wall, which were obtained during subsequent surgical treatment of GP. Methods of parametric statistics were used.&#x0D; Results. It was established that focal dystrophy, acanthotic severity, keratinization of the vaginal epithelium, moderate inflammation are characteristic morphological changes in the vaginal mucosa in women with postmenopausal GP. Application at the stage of preparation for surgical treatment using vaginal forms, as a result of which estrogen and progesterone enhance the inflammatory process in the ulcerative vagina, reduce dystrophic changes and their severity in relation to complex changes and differentiation of the vaginal epithelium and activation of regeneration processes in tissues.&#x0D; Conclusion. Reducing inflammation and the severity of dystrophic processes in vaginal enemas at the stage of preoperative preparation of women with postmenopausal GP favorably affects the regeneration processes, which in the first place can improve the outcome of surgical treatment.

Variation in outcome reporting and measurement tools in clinical trials of treatments for genitourinary symptoms in peri- and postmenopausal women: a systematic review.

Genitourinary symptoms affect 40% to 60% of postmenopausal women. Evidence-based approaches to diagnosing and managing these symptoms are limited by inconsistencies in outcomes and measures used in clinical trials. The aim of the study was to systematically review all outcomes and measurement tools reported in randomized clinical trials of interventions for genitourinary symptoms associated with menopause. We searched PubMed, Scopus, and the Cochrane Central Register of Controlled Trials (CENTRAL) up to December 2018. Randomized controlled trials with a primary or secondary outcome of genitourinary symptoms associated with menopause, English language, and sample size of 20 or more women per study arm were included. Study characteristics, outcomes, and measurement methods were collected. The search yielded 3,478 articles of which 109 met inclusion criteria. Forty-eight different outcomes were reported with "atrophy" as the most common (56/109, 51%) followed by measures of sexual function (19/109, 17%). Almost all (108/109, 99%) trials included patient-reported measures, with 21 different measures and 39 symptom combinations. Clinician-reported scales of vulvovaginal appearance were used in 36 of 109 (33%) trials, with extensive variation in what was measured and reported. Cytological measures from the vaginal epithelium were the most commonly used objective tools (76/109, 70%). There is heterogeneity in reported outcomes and measures used in clinical trials of treatments for genitourinary symptoms at menopause and uncertainty as to which outcomes best reflect patient priorities and symptoms. The findings from this systematic review have informed an international survey of stakeholders to determine priorities for outcome selection and reporting. This survey will then inform the development of a Core Outcome Set for use in future clinical trials by the COMMA (Core OutcoMes in MenopAuse) consortium. : Video Summary:http://links.lww.com/MENO/A599.

OVARIAN AGING AND REPRODUCTIVE SENESCENCE IN MOUSE MODEL OF MITOCHONDRIAL DYSFUNCTION

To evaluate the mechanisms of female reproductive aging and senescence induced by mitochondrial dysfunction in mice. Prospective cohort design study. 13 female mice with mitochondrial dysfunction (mtDNA-depleter) and 13 wild-type (WT) female mice were studied at three time points in their reproductive life cycle: 4-6, 8-10, and 11-12 months of age. A transgenic mouse model with an inducible gene for mitochondrial dysfunction (mtDNA-depleter) was previously created by our lab. All protocols were approved by our institutional IACUC. At each of the three time points, reproductive markers were assessed, including estrous cyclicity, plasma anti-Mullerian hormone (AMH), ovarian follicles, estrogen receptor staining, and uterine and vaginal atrophy. Breeding experiments were performed to assess fertility. Differences between the mtDNA-depleter and WT groups were evaluated by independent two-sample t-test. Twenty-six female mice were studied from 4 to 12 months of age. The estrous cycles in 4-6-month mtDNA-depleter mice were prolonged, 9.4 days (n = 7), versus 5.3 days in WT (n = 9), p-value of 0.021. By 11-12 months, both groups had prolonged estrous cycles consistent with physiologic aging. AMH levels in 4-6-month mtDNA-depleter mice were decreased, 84 (n = 4), versus 114 ng/mL in WT (n = 5), p-value of 0.013. AMH was reduced in older mice at 8-10 and 11-12 months, but there were no significant differences between groups. Looking at the data collectively for all three time points, there were fewer tertiary (early antral, antral, or pre-ovulatory) follicles in mtDNA-depleter mice than in WT, 3.5 versus 4.9 per cut section, respectively (p-value of 0.039). There were fewer corpora lutea in mtDNA-depleter mice than in WT, 0.90 versus 1.7 per cut section, respectively (p-value of 0.027). Collectively, average uterine weight in mtDNA-depleter mice was lower, 62.2, versus 92.3 mg in WT mice, p-value of 0.0028. Vaginal epithelium in 4-6-month mtDNA-depleter mice was thinner, 98 μm (n = 4), versus 200 μm in WT mice (n = 4), p-value of 0.035. In breeding experiments, mtDNA-depleter mice at age 3 and 5 months did not produce litters. Estrogen receptor levels were reduced in mtDNA-depleter mouse ovary relative to WT, evident from both immunofluorescence and semi-quantitative PCR analysis. Mitochondrial dysfunction in mice is associated with premature reproductive aging and senescence, as evidenced by early onset of prolonged estrous cycles, diminished ovarian reserve, uterine and vaginal atrophy, and sterility. Mitochondrial dysfunction is also associated with premature estrogen receptor down-regulation in the mouse ovary.