Study question Is there any correlation between the total bacterial load and the lactobacilli quantities in the vaginal and endometrial microbiomes in reproductive-age women? Summary answer There was no correlation between the vaginal and endometrial total bacterial loads and only a weak positive correlation between the quantities of lactobacilli. What is known already The Lactobacilli-dominated microbiota is considered to be the most favorable type of microbiota in the uterine cavity. It is associated with increased reproductive success in women undergoing in vitro fertilization. Whereas the non-Lactobacillus dominated microbial communities are more frequent in women with poor pregnancy outcomes. When analyzing endometrial microbiota, one of the challenges is sampling. Transvaginal sample intake involves the possibility of contaminating the samples with vaginal microbiota. Moreover, it is an invasive procedure leading to the development of infectious inflammatory diseases of the upper genital tract. Thus, researchers are currently searching for predictors of the state of endometrial microbiota. Study design, size, duration It is a cross-sectional study of the vaginal end endometrial micorbiomes from 64 reproductive-age women. Endometrial and vaginal samples were collected simultaneously on days 7–10 of the menstrual cycle. To avoid contamination by vaginal microbiota, Endobrush Standard for Endometrial Cytology (Laboratoire C.C.D.; France) was used for endometrial sampling. Participants/materials, setting, methods The study included women who came to the “Garmonia” Medical Center (Yekaterinburg, Russia) seeking infertility treatment. The average age of the patients was 32.2±5.0. DNA from vaginal and endometrial samples was extracted using PREP-NA-PLUS kit (DNA-Technology, Russia). Vaginal and endometrial microbiota was analyzed using Femoflor real-time PCR kit and DTprime 4M1 thermocycler (DNA-Technology, Russia). Main results and the role of chance Total bacterial load (TBL) in vaginal discharge was 3.8–7.9 lg (median — 7.1, interquartile range — 6.6–7.4). TBL in the endometrial samples was 0–5.1 lg (median — 3.9, interquartile range — 3.6–4.2). There was no correlation between TBL values in vaginal discharge and endometrial samples (Spearman’s rho — 0.247, p = 0.049). Lactobacilli quantities in vaginal discharge were 4.5–8.3 lg (median — 7.2, interquartile range — 6.4–7.6), in endometrial samples — 0–5.1 lg (median — 3.7, interquartile range — 3.1–4.2). There was a weak positive correlation between lactobacilli quantities in vaginal and endometrial samples (Spearman’s rho — 0.362, p = 0.003). The proportion of lactobacilli in vaginal discharge was 1–100% (median — 100%, interquartile range — 95–100%), in the endometrial samples — 0–100% (median — 96%, interquartile range — 25–100%). There was no correlation between lactobacilli proportions in vaginal and endometrial samples (Spearman’s rho — 0.225, p = 0.074). Furthermore, there was no correlation between lactobacilli quantity in the vagina and their proportion in the endometrial microbiota (Spearman’s rho — 0.294, p = 0.018). There was only a weak positive correlation between the quantities of lactobacilli in vaginal and endometrial samples. Vaginal TBL values and lactobacilli proportions did not correlate with lactobacilli quantities and proportions in the endometrial samples. Limitations, reasons for caution The study was conducted on a small sample. Moreover, it is notoriously difficult to interpret the analysis results for endometrial microbiota due to the high risk of contamination and its low microbial biomass. Wider implications of the findxings: Apparently, there is no obvious link between the vaginal and endometrial microbiomes. It is possible that, apart from vaginal microbiota, there are other predictors which could allow us to assume whether lactobacilli are present in the endometrial microbiota. Trial registration number Not applicable
Introduction. Normal ovarian reserve (OR) determining the ovarian response to follicle development containing fully-featured oocytes is an important factor in pregnancy, including assisted reproductive technology (ART) programs. The causes of premature OR decrease are multifactorial. The study of gene polymorphism as a cause of the premature OR decrease deserves attention.Aim: to determine genetic predisposition to premature OR decrease and create a prognostic model based on study results.Materials and Methods. A retrospective comparative cohort study was conducted. A total of 200 reproductively active patients with infertility underwent ART were examined. The patients were divided into 2 groups: Group 1 included 100 patients with a premature OR decrease; Group 2 consisted of 100 patients with a normal OR. All patients underwent molecular genetic study. Genetic polymorphisms of the genes ESR1, ESR2, FSHR, CYP19A were studied.Results. The final clinical phenotype is shaped by multiple factors – genetic and environmental. Several genetic variants contribute to the formation of premature decrease in ovarian reserve. Gene combination of CYP19A1 and FSHR displayed the greatest synergistic effect, potentiating each other and predisposing to a poor ovarian response as part of stimulated superovulation.Conclusion. Identifying genetic markers is a promising method for individual OR evaluation, including its premature decline. Multilocus analysis and a prognostic model based on combining several polymorphic gene variants will allow to assess risks of premature OR decrease and individualize ART programs.
Introduction. In the structure of gynecological morbidity intrauterine pathology occupies a leading position because of its high frequency and negative impact on the reproductive health of a woman. Endometrial pathology is numerous and includes endometrial hyperplasia and polyps, endometrial cancer, abnormal uterine bleeding and dysmenorrhea, endometriosis and Asherman's syndrome. Pathological changes of the endometrium in women with reproductive problems, such as infertility, habitual miscarriage, are particularly relevant.The aim of the work is to estimate the contribution of endometrial microbiota in the development of endometrial pathological processes and identify possible pathogens for the development of this process on the basis of scientific literature data.Materials and methods. Data analysis and systematization were performed on the basis of scientific publications in the PubMed, Google scholar, and Scopus databases for the period 2010–2021. For the final scientific review 65 sources were selected.Results and discussion. This article presents findings on the role of the uterine cavity microbiota and its contribution to endometrial pathology such as endometrial polyps and hyperplasia, adenomyosis and uterine bleeding, infertility and intrauterine synechiae. The correction of an altered microbiota opens up new insights into the development of uterine diseases and new therapeutic options.Conclusion. Numerous studies of the uterine cavity microbiota show its undoubted role in the development of uterine cavity pathology. However, the data obtained are quite contradictory. Further study of the uterine microbiota is necessary for a deeper understanding of the pathogenesis of endometrial pathological processes.
Study question Can we find specific biomarkers in vaginal microbiota predicting the state of endometrial microbiota? Summary answer The higher levels of Eubacterium spp. in the vagina could serve as a predictor of the non-Lactobacillus-dominated microbial community in the uterus. What is known already The absence of microorganisms or the Lactobacillus-dominated community in the uterus cavity is considered more favorable than the non-Lactobacillus-dominated community. The latter is associated with poor pregnancy outcomes in women undergoing in vitro fertilization. However, endometrial microbiota analysis is challenging due to the possibility of contaminating the samples with vaginal microbiota during transvaginal sampling. Also, there is always a risk of upper genital tract inflammation after the sampling. For these reasons, the idea of finding markers predicting the state of endometrial microbiota is promising. Study design, size, duration It is a cross-sectional study of the vaginal and endometrial microbiota in 61 reproductive-age women with the normal histological pattern of endometrial samples. Endometrial and vaginal samples were collected simultaneously on days 7–10 of the menstrual cycle. Depending on the type of endometrial microbiota, the women were divided into two groups: Group 1 – the Lactobacillus-dominated microbiota or no microbiota (n = 43), Group 2 – the non-Lactobacillus-dominated microbiota (n = 18). Participants/materials, setting, methods All women came to the “Garmonia” Medical Center (Yekaterinburg, Russia) seeking infertility treatment from 2019 to 2021. Vaginal and endometrial microbiota was analyzed using Femoflor and Androflor real-time PCR kits (DNA-Technology, Russia). For the vaginal samples, the quantities and proportions of 22 microorganism groups were compared in Group 1 and Group 2 using Mann-Whitney test. For the variables with significant differences, ROC curves were constructed to assess the diagnostic value of the tests. Main results and the role of chance When comparing vaginal microbiota in Groups 1 and 2, we registered the statistically significant differences for 6 out of 44 variables (quantities and proportions of 22 microorganism groups). These variables were the quantities of Eubacterium spp. (10° vs. 104.7, p = 0.002) and Gardnerella vaginalis (10° vs. 104.0, p = 0.033), the proportions of obligate anaerobes (0% vs. 6.0%, p = 0.010), Peptostreptococcus spp./Parvimonas spp. (0% vs. 0%, p = 0.011), G. vaginalis (0% vs. 0.2%, p = 0.012), Eubacterium spp. (0% vs. 0.6%, p = 0.033). “Eubacterium spp.’ quantity” variable demonstrated the highest discriminatory power (the area under the ROC curve value — 0.737 [CI 0.593-0.882], p = 0.004). The AUC value was considered as acceptable to divide patients into Group 1 and Group 2. The cutoff value of Eubacterium spp. 103.15 genome-equivalent/ml (GE/ml)) allowed to predict the non-Lactobacillus-dominated microbiota in the uterus cavity with 77.8% sensitivity and 62.8% specificity. Limitations, reasons for caution The results were obtained on a small population group of women with the normal histological pattern of endometrial samples. It may be different if the larger pool of samples is investigated. It is also possible, that the results will not apply to the patients with endometrial abnormalities. Wider implications of the findings Further research should determine the diagnostic value of Eubacterium spp. identified in vaginal samples as the predictor for the endometrial microbiota composition in women with endometrial abnormalities. The next step is research dedicated to the link between different signatures in vaginal microbiota and endometrial abnormalities. Trial registration number Protocol № 7 dated September 20, 2019
Chronic endometritis (CE) in women of the reproductive age is associated with infertility and recurrent pregnancy loss. The aim of this study was to evaluate the endometrial microbiota by means of real-time PCR in reproductive-age women depending on the morphological pattern of the endometrium. Using the Androflor real-time PCR kit, we analyzed endometrial aspirate collected from 23 patients with chronic endometritis, 30 patients with endometrial hyperplasia, and 19 healthy women. DNA of up to 9 groups of microorganisms was detected in all the analyzed samples in the amounts exceeding negative control. The total bacterial load (TBL) of the detected microorganisms was 10<sup>3</sup>–10<sup>6,4</sup> (median 10<sup>3,8</sup>) GE/ml. Lactobacillus spp. were detected the most often (86.1% of all samples). Opportunistic microorganisms (OM) were identified in 36.1% of all samples, including 22.2% of samples with lactobacilli and 13.9% — without lactobacilli. The variant of microbiota composition with Lactobacillus-dominance (more than 90%. in the TBL) was detected significantly less often in women with chronic endometritis compared to healthy women. Real-time PCR could be used for assessment of endometrial microbiota and allows us to determine its characteristics depending on the morphological pattern.
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