P–522 Cervical secretion methylation profile is associated with the success of frozen-thawed embryo transfer - a proof-of-concept study

Study question Is cervical secretion gene methylation profile different between receptive and non-receptive endometrium and associated with implantation outcome in frozen-embryo transfer (FET) cycle? Summary answer The combination of candidate genes methylation profiles obtained from cervical secretion showed significant associations with pregnancy outcomes. What is known already Implantation failure remains a black box in reproductive medicine, and the exact mechanism of how endometrial receptivity is regulated is still unknown. Epigenetic modifications play a role in the gene expression pattern and may alter the endometrial receptivity in the human endometrium. Cervical secretion containing various implantation-related cytokines, and the gene methylation change can be used as a non-invasive molecular source that reflects the endometrium condition. Study design, size, duration In this retrospective case-control study, sixty-two women who entered the FET cycle (30 pregnant and 32 non-pregnant women) were enrolled. Participants/materials, setting, methods Cervical secretion was collected before embryo transfer from women enrolled in multicenter university-affiliated reproductive units. The DNA methylation status of six candidate genes was measured using quantitative methylation-specific PCR (qMSP). The correlation between methylation change and the pregnancy outcome was analyzed. Main results and the role of chance The candidate genes were selected from that associated with implantation with literature review and the original genome-wide DNA methylation data from NCBI GEO DataSets (GSE90060) which processed using bioinformatics analysis. Six candidate genes whose CpG-level methylation analysis with β-value statistically higher in receptive endometrium than in a pre-receptive endometrium were selected. All six candidate genes showed different degrees of correlation with the pregnancy outcomes. Among them, PRKAG2 methylation changes showed the highest correlation with the pregnancy outcome. A logistic regression model was used to evaluate the performance of a single gene or a combination of genes for implantation prediction. The results showed a statistically significant association between the methylation status of a combination of genes (PRKAG2, KRS1, HAND2) and the pregnancy outcome (p = 0.008), resulting in an optimal AUC of 0.7 (95% CI: 0.57 - 0.81) for implantation prediction. Limitations, reasons for caution The results obtained from a relatively small cohort size. A larger study and further comprehensive methylome investigations are warranted. Wider implications of the findings: This study is the first proof-of-concept study that cervical secretion methylation profile is associated with implantation outcome in a FET cycle, and showed potential as a non-invasive method for implantation prediction. Trial registration number non applicable

Congenital, Intrapartum and Postnatal Maternal-Fetal-Neonatal SARS-CoV-2 Infections: A Narrative Review

Background: There is inconclusive evidence regarding congenital, intrapartum, and postnatal maternal-fetal-neonatal SARS-CoV-2 infections during the COVID-19 pandemic. A narrative review was conducted with the aim of guiding clinicians on the management of pregnant women with respect to congenital, intrapartum, and postnatal maternal-fetal-neonatal SARS-CoV-2 infections and breastfeeding during the COVID-19 pandemic. Methods: Searches were conducted in Web of Science, PubMed, Scopus, Dialnet, CUIDEN, Scielo, and Virtual Health Library to identify observational, case series, case reports, and randomized controlled trial studies assessing the transmission of SARS-CoV-2 from mother to baby and/or through breastfeeding during the COVID-19 pandemic. Results: A total of 49 studies was included in this review, comprising 329 pregnant women and 331 neonates (two pregnant women delivered twins). The studies were performed in China (n = 26), USA (n = 7), Italy (n = 3), Iran (n = 2), Switzerland (n = 1), Spain (n = 1), Turkey (n = 1), Australia (n = 1), India (n = 1), Germany (n = 1), France (n = 1), Canada (n = 1), Honduras (n = 1), Brazil (n = 1), and Peru (n = 1). Samples from amniotic fluid, umbilical cord blood, placenta, cervical secretion, and breastmilk were collected and analyzed. A total of 15 placental swabs gave positive results for SARS-CoV-2 ribonucleic acid (RNA) on the fetal side of the placenta. SARS-CoV-2 RNA was found in seven breastmilk samples. One umbilical cord sample was positive for SARS-CoV-2. One amniotic fluid sample tested positive for SARS-CoV-2. Conclusions: This study presents some evidence to support the potential of congenital, intrapartum, and postnatal maternal-fetal-neonatal SARS-CoV-2 infections during the COVID-19 pandemic. Mothers should follow recommendations including wearing a facemask and hand washing before and after breastfeeding.

Persistência do HPV em mulheres tratadas para o adenocarcinoma cervical

Objetivo: determinar a frequência da persistência do HPV em mulheres tratadas para o adenocarcinoma cervical. Método: trata-se de um estudo quantitativo, descritivo, retrospectivo, do tipo coorte, com 77 mulheres genotipadas para o HPV antes do tratamento e com adenocarcinoma cervical. Coletou-se novo material de secreção cervical após o tratamento para a realização da detecção do DNA do HPV por reação de cadeia da polimerase. Utilizou-se o programa estatístico Epi Info 7.1.4. para a análise dos dados. Resultados: observou-se que, das 77 pacientes, foi possível determinar a genotipagem do HPV após o tratamento em 30 mulheres e, destas, 7 (23,3%) apresentaram o HPV detectável. Encontraram-se os tipos de HPV dos quais quatro pacientes (57,1%) estavam com o HPV 31, sendo que, em uma paciente, estava associado ao HPV 18; o 33 em duas mulheres (28,6%), sendo que em uma estava associado ao HPV 16 e uma apresentou os HPV 11 e 56 associados (14,2%). Conclusão: detectou-se o HPV na secreção cervical em 23,3% das mulheres após o tratamento para o adenocarcinoma de colo uterino, sendo o HPV 31 o tipo mais frequente. Descritores: Adenocarcinoma; HPV; Falha de Tratamento; Prevalência; Colo do Útero; Doenças Sexualmente Transmissíveis. AbstractObjective: to determine the frequency of persistence of HPV in women treated for cervical adenocarcinoma. Method: this is a quantitative, descriptive, retrospective, cohort study, with 77 women genotyped for HPV before treatment and with cervical adenocarcinoma. New cervical secretion material was collected after treatment to detect HPV DNA by polymerase chain reaction. The Epi Info 7.1.4 statistical program was used for data analysis. Results: it was observed that, out of 77 patients, it was possible to determine HPV genotyping after treatment in 30 women, and of these, seven (23.3%) had detectable HPV. HPV types were found, of which four patients (57.1%) had HPV 31, and in one patient it was associated with HPV 18; o 33 in two women (28.6%), one of whom was associated with HPV 16 and one had HPV 11 and 56 associated (14.2%). Conclusion: HPV was detected in cervical secretion in 23.3% of women after treatment for cervical adenocarcinoma, with HPV 31 being the most frequent type. Descriptors: Adenocarcinoma; Papillomaviridae; Treatment Failure; Prevalence; Cervix Uteri; Sexually Transmitted Diseases.ResumenObjetivo: determinar la frecuencia de persistencia del VPH en mujeres tratadas por adenocarcinoma cervical. Método: estudio de cohorte cuantitativo, descriptivo, retrospectivo, con 77 mujeres genotipadas para el VPH antes del tratamiento y con adenocarcinoma cervical. Se recogió nuevo material de secreción cervical después del tratamiento para detectar el ADN del VPH por reacción en cadena de la polimerasa. Se utilizó el programa estadístico Epi Info 7.1.4. para el análisis de datos. Resultados: se observó que, de las 77 pacientes, fue posible determinar el genotipo del VPH después del tratamiento en 30 mujeres, y de estos, 7 (23.3%) tenían VPH detectable. Se encontraron tipos de VPH, de los cuales cuatro pacientes (57.1%) tenían VPH 31, y en una paciente se asoció con VPH 18; o 33 en dos mujeres (28,6%), una de las cuales estaba asociada con el VPH 16 y una tenía VPH 11 y 56 (14,2%). Conclusión: el VPH se detectó en la secreción cervical en el 23,3% de las mujeres después del tratamiento para el adenocarcinoma de cuello uterino, siendo el VPH 31 el tipo más frecuente. Descriptores: Adenocarcinoma; Papillomaviridae; Insuficiencia del Tratamiento; Prevalencia; Cuello del Útero; Enfermedades de Transmisión Sexual.

Low intensity laser effects on factors of congenital immunity and cytokine balance in secretions of female reproductive system affected with chlamydia infection

Therefore, we defined the objective of our research: to study the factors of uterine cervix tissue im- munity for women with Chlamydia infection and to estimate the effect of low-intensity laser therapy (LILT) on these factors due to a wavelength of 632 nm and a power of 30 mW. Materials and methods. Clinical immunologic studies of cervical secretion were carried out on 184 women of reproductive age 25,9 ± 4,22 in 2016–2018. Chlamydia infection Chlamydia trachomatis was found by the methods of real-time polymerase chain reaction (Rt-PCR) tests. Code by Classifier of the International Standard Classification of Diseases and Related Health Problems 10th Revision is А 56.0, Chlamydial infections of the lower sections of the urogenital tract. Urethra and canalis cervicis uteri scrapings were used as the material for detecting C. trachomatis. All patients with chlamydial infection of the lower part of the urogenital system were divided into two groups: a group of 100 women who were treated by using the local influence of the low-intensity laser of 632 nm as a part of complex treatment, the comparison group (basic treatment) consisted of 84 women who took azithromycin of 1 g once a day for the 1st, 5th, 10th days of treatment. 50 healthy women without any pathology and infections of the genitourinary system made the control group. A system of cytokines was studied by interleukin's level IL-8. IL-1 α, IL-1 β, TNF-α and IFN-γ content in cervical mucus. Results Course. The results of the study showed a high clinical and immunological efficacy of treating chlamydial cervicitis using a low intensity laser with a wavelength of 632 nm. An imbalance of cellular factors of innate immunity revealed before starting a therapy: a decrease in the oxygen-dependent metabolism of neutrophilic granulocytes of the cervical secretion.

New Possibilities in Molecular Diagnosis and Prognosis of Diseases Associated with Human Papilloma Virus in Cervical Secretion

The aim of the article is to determine the importance of the systematic analysis of data available in modern publications – molecular markers of cervical discharge in the diagnosis of various diseases of the cervix associated with human papilloma virus (HPV). In addition, the prospects of studying protein and metabolic profiles of cervicovaginal discharge for predicting the course of neoplastic processes of the cervix. The article describes a number of protein markers in cervical defined for the early diagnosis of precancerous diseases and cervical cancer.

Cytokine profile of cervical mucus at different forms of papillomavirus infection

Aim. To study the levels of pro- and anti-inflammatory cytokines in cervical secretion in females with cervical epithelial dysplasia and latent papillomavirus infection. Methods. The study included 120 females aged 20 to 40 years with cervical papillomavirus infection, who were assigned to two groups using «case-control» method. The first group included 60 females with latent disease, the second group - 60 females with mild and moderate cervical epithelial dysplasia (cervical intraepithelial neoplasia stages I and II). Cytokine levels in cervical secretion were measured by ELISA using «ProCon» test system. Results. ELISA test showed increased levels of interleukin-4 and interleukin-10 (43 [21; 74] and 48 [12; 88] pg/ml, respectively) and decreased levels of interleukin-2 (18.5 [5.5; 27.5] pg/ml), interleukin-6 (0.6 [0.06; 0.9] pg/ml), tumor necrosis factor alpha (88.5 [0; 123] pg/ml), interferon γ (2 [0; 4] pg/ml) in cervical secretion of females with cervical epithelial dysplasia compared to females with latent papillomavirus infection. Thus, females with cervical epithelial dysplasia showed increased levels of anti-inflammatory cytokines: interleukin-4 - by 2.7 times and interleukin-10 - by 2.4 times compared to females with latent disease, while levels of pro-inflammatory cytokines was decreased as following: interleukin-2 - by 1.4 times, interleukin-6 - by 4.5 times, tumor necrosis factor alpha - by 1.8 times, interferon γ - by 6.3 times (p 0.05). Conclusion. Imbalance of immune response cytokine regulation with anti-inflammatory cytokines prevailing might be an important factor facilitating persistence of papillomavirus in cervical epithelium and contributing to cervical epithelial dysplasia onset and progression.