Long pentraxin 3 and vitamin D receptor mRNA expression pattern of cumulus granulosa cells isolated from PCOS oocytes at different stages of nuclear maturation.

BACKGROUND: A fine-tuned pro-inflammatory and anti-inflammatory balance in the follicular unit is essential for cumulus expansion and successful ovulation. While the long pentraxin 3 (PTX3) gene is required for the expansion of cumulus cells (CCs), ovulation, resumption of meiosis and fertilization, the vitamin D receptor gene (VDR-X2) is required for intra-follicle redox balance. This study was planned to determine the expression pattern of VDR-X2 and PTX3 mRNA in CCs isolated from germinal vesicle (GV), metaphase I (MI), and metaphase II (MII) oocytes of PCOS patients with ovulatory dysfunction. METHODS: The relative expression of CC-PTX3 and CC-VDR-X2 mRNA were evaluated using qRT-PCR in a total of 79 CC samples collected from individual cumulus-oocyte complex of 40 infertile patients (20 PCOS and 20 non-PCOS normal responders) who underwent ovarian stimulation with the GnRH antagonist protocol. RESULTS: Relative PTX3 mRNA expressions of CCMI-control and CCMII-control showed 3- and 9-fold significant upregulation compared to CCGV-control, respectively. The relative PTX3 mRNA expression of CCMII-control increased approximately three fold compared to CCMI-control. Compared to CCGV-pcos, a 3-fold increase was noted in the relative PTX3 mRNA expression of CCMI-pcos and an approximately 4-fold increase in the PTX3 mRNA expression of CCMII-pcos. Relative PTX3 mRNA expression values of CCMII-pcos and CCMI-pcos were similar. A 6-fold upregulation of relative PTX3 mRNA and a 4-fold upregulation of VDR-X2 mRNA were detected in CCMII-control compared to CCMII-pcos. CC-VDR-X2 expression patterns of the PCOS and control groups overlapped with the CC-PTX3 pattern. Fertilization rates of the PCOS group exhibiting failed transcript expression were similar to normal responders. CONCLUSION: The fact that relative CC-PTX3 and CC-VDR mRNA expression does not increase during the transition from MI to MII stage in PCOS as in normal responders suggests that PTX3 and VDR expression may be defective in cumulus cells of PCOS patients with ovulatory dysfunction.

Endometrial Injury Upregulates Expression of Receptivity Genes in Women with Implantation Failure.

BACKGROUND: Homeobox genes A10 (HOXA10) and A11 (HOXA11), members of the abdominal B gene family, are responsible for embryonic survival and implantation. This study was planned to investigate whether endometrial injury alters the expression of both transcripts in women with implantation failure. METHODS: A total of 54 women with implantation failure were divided into two equal groups as experimental (scratching) and sham (no scratching). Participants in the scratching group were exposed to endometrial injury in the mid-luteal phase, and those in the sham group were exposed to endometrial flushing. The scratching group, but not the sham group, underwent prior endometrial sampling. A second endometrial sampling was performed on the scratching group in the mid-luteal phase of the following cycle. The mRNA and protein levels of the HOXA10 and 11 transcripts were determined in endometrial samples collected before and after injury/flushing. Participants in each group underwent IVF/ET in the cycle after the second endometrial sampling. RESULTS: Endometrial injury caused a 60.1-fold (p < 0.01) increase in HOXA10 mRNA and a 9.0-fold increase in HOXA11 mRNA (p < 0.02). Injury resulted in a significant increase in both HOXA10 (p < 0.001) and HOXA11 protein expression (p < 0.003). There was no significant change in HOXA10 and 11 mRNA expressions after flushing. Clinical pregnancy, live birth, and miscarriage rates of the both groups were similar. CONCLUSIONS: Endometrial injury increases homeobox transcript expression at both mRNA and protein levels.

Endometrial injury concurrent with hysteroscopy increases the expression of Leukaemia inhibitory factor: a preliminary study.

OBJECTIVE: It is not known by which mechanism endometrial injury increases pregnancy rates. Leukaemia inhibitory factor (LIF) is a cytokine involved in wound healing and implantation. The aim of this study was to determine the change in endometrial LIF mRNA expression before and after mechanical injury during hysteroscopy. METHODS: Forty patients with a history of two or more unsuccessful implantations who decided to undergo hysteroscopy in the proliferative phase were divided into two equal groups: one with endometrial injury (scratching group) and the other with noninjury (control group). Endometrial sampling was conducted before injury on the patients in the scratching group, and then injury was performed with monopolar needle forceps. Only diagnostic hysteroscopy was performed on the patients in the control group. Endometrial tissues were collected using a Pipelle catheter between Days 20 and 23 of the mid-luteal phase of the next cycles in both the scratching and control groups. Endometrial LIF mRNA expression was evaluated with the use of reverse-transcription polymerase chain reactions. RESULTS: Relative changes in mRNA expression levels of the LIF gene in endometrial samples taken before and after injury were calculated using the 2-ΔΔCt method, and the fold changes obtained were compared between and within the groups. Compared with preinjury values, an 11.1-fold increase was found in postinjury LIF mRNA expression in patients with monopolar forceps injury (p < 0.001). There was a 3.9-fold significant increase in postinjury LIF mRNA levels compared with those in the control group (p < 0.02). CONCLUSIONS: The fertility-promoting effect of hysteroscopy-guided mechanical endometrial injury may be mediated by LIF mRNA.

Factors preventing materno-fetal transmission of SARS-CoV-2.

Although many pregnant women have been infected by coronavirus, the presence of intrauterine vertical transmission has not been conclusively reported yet. What prevents this highly contagious virus from reaching the fetus? Is it only the presence of a strong placental barrier, or is it the natural absence of the some receptor that the viruses use for transmission? We, therefore, need to comprehensively understand the mechanism of action of the mammalian epithelial barriers located in two different organs with functional similarity. The barriers selected as potential targets by SARS-CoV-2 are the alveolo-capillary barrier (ACB), and the syncytio-capillary barrier (SCB). Caveolae are omega-shaped structures located on the cell membrane. They consist of caveolin-1 protein (Cav-1) and are involved in the internalisation of some viruses. By activating leukocytes and nuclear factor-κB, Cav-1 initiates inflammatory reactions. The presence of more than one Cav-1 binding sites on coronavirus is an important finding supporting the possible relationship between SARS-CoV-2-mediated lung injury. While the ACB cells express Cav-1 there is no caveolin expression in syncytiotrophoblasts. In this short review, we will try to explain our hypothesis that the lack of caveolin expression in the SCB is one of the most important physiological mechanisms that prevents vertical transmission of SARS-CoV-2. Since the physiological Cav-1 deficiency appears to prevent acute cell damage treatment algorithms could potentially be developed to block this pathway in the non-pregnant population affected by SARS-CoV-2.

Combating sars-cov-2 through lipoxins, proteasome, caveolin and nuclear factor-κb pathways in non-pregnant and pregnant populations.

It can be misleading to think that the new severe acute respiratory syndrome coronavirus (SARS-CoV2) which has a very strong mutation and adaptation capabilities, uses only the angiotensin-converting enzyme II (ACE2) pathway to reach target cells. Despite all the precautions taken, the pandemic attack continues and the rapid increase in the number of deaths suggest that this virus has entered the cell through different pathways and caused damage through different mechanisms. The main reason why the ACE2 pathway comes to the fore in all scientific studies is that this receptor is located at the entry point of basic mechanisms that provide alveolo-capillary homeostasis. SARS-CoV-2 has to use nuclear factor-κB (NF-kB), caveloae, clathrin, lipoxin, serine protease and proteasome pathways in addition to ACE2 to enter the target cell and initiate damage. For this reason, while new drug development studies are continuing, in order to be beneficial to patients in their acute period, it is imperative that we are able to come up with drugs that activate or inhibit these pathways and are currently in clinical use. It is also critical that we adopt these new pathways to the treatment of pregnant women affected by SARS-CoV-2, based on the scientific data we use to treat the general population.

Surgical Removal of Hydrosalpinx Improves Endometrium Receptivity by Decreasing Nuclear Factor-Kappa B Expression.

Although physiologic amount of inflammation is necessary for successful implantation, pathological inflammation inhibits the expression of receptivity molecules and genes. Because hydrosalpinges is an inflammatory disease, adverse effects of hydrosalpinges on implantation may be in part mediated by disturbed endometrial expression of nuclear factor-kappa B (NF-κB), a dimer implicated in inflammation. We examined the expression of NF-κB p65 (Rel A) during the window of implantation in the endometrium of infertile women (n = 14) with uni or bilateral hydrosalpinges prior to and following salpingectomy and of fertile controls (n = 14) by immunohistochemistry. We assessed the influence of salpingectomy on NF-κB p65 expression by comparing pre- and post-operative endometrial samples. To evaluate the intensity of endometrial NF-κB p65 (Rel A) immunoreactivity, H-score method was used. We showed a decrease in NF-κB p65 expression in 13 out of the 14 post-salpingectomy endometrial samples. The mean NF-κB p65 level was significantly higher in the endometrium of women with hydrosalpinges before salpingectomy compared with control cases without hydrosalpinges (3.94 ± 6.2 vs 2.18 ± 0.7, p < .02). Salpingectomy decreased the mean endometrial NF-κB p65 levels in both unilateral and bilateral hydrosalpinges (2.87 ± 1.1). When we compared the endometrial NF-κB p65 levels of the post-salpingectomy samples with their age-matched fertile controls, we did not observe any significant difference. After salpingectomy, the mean H-score of endometrial NF-κB p65 expression significantly decreased to a level similar to that of the fertile group (2.87 ± 1.1 vs 2.18 ± 0.7, p > .64). NF-κB p65 expression was detected in cytoplasmic and membranous parts of luminal and glandular epithelial cells of endometrium obtained before salpingectomy. Both epithelial and stromal components of the endometrium showed decreased staining for NF-κB p65 compared with the pre- and post-salpingectomy samples. The decreased NF-κB p65 (Rel A) immunoreactivity was predominantly localized to luminal and glandular epithelial cells. Uni or bilateral hydrosalpinges causes pathological endometrial inflammation. Improvement in pathological inflammation following salpingectomy in women with hydrosalpinges may be in part mediated by the downregulation of endometrial NF-κB p65 (Rel A) expression.

Testis spectroscopy may predict sperm retrieval rate in men with non-obstructive azoospermia undergoing micro-TESE: A pilot study

Objective: To investigate whether prior testis magnetic resonance spectroscopy predicts the success or failure of micro-dissection testicular sperm extraction (micro-TESE) in patients with non-obstructive azoospermia (NOA). Material and Methods: Nine men with NOA who were scheduled for micro-TESE for the first time, 9 NOA men with a history of previous micro-TESE and 5 fertile men were enrolled. All NOA patients and fertile controls underwent testis spectroscopy. A multi-voxel spectroscopy sequence was used. Testicular signals of choline (Cho), creatine (Cr), myo-inositol (MI), lactate, and lipids were analyzed quantitatively and compared with the results of the micro-TESEs. Results: The most prominent peaks were Cho and Cr in the fertile controls and NOA subjects with positive sperm retrieval in the micro-TESE. A high Cho peak was detected in 87% of the NOA men with positive sperm retrieval. NOA men without sperm at the previous micro-TESE showed a marked decrease in Cho and Cr signals. For positive sperm retrieval in micro-TESE, the cut-off value of Cho was 1.46 ppm, the cut-off value of Cr was 1.43 ppm, and the cut-off value of MI was 0.79 ppm. Conclusion: Testis spectroscopy can be used as a non-invasive screening method to predict the success or failure of micro-TESE.

Human papillomavirus prevalence and genotype distribution among Turkish women with or without cervical lesion.

CONTEXT: Human papillomavirus (HPV) infection is the main cause of cervical cancer, but the risk is associated with the various HPV genotypes which may be found in women with or without clinical findings. AIMS: We aimed to identify HPV prevalence and genotype distribution in women with or without cervical lesions admitted to Gynaecology and Obstetrics Clinics of one of the largest private hospitals in Istanbul between 2013 and 2017. SUBJECTS AND METHODS: In the present study, cervical cytobrush samples collected from 2464 women with different cytological conditions, and investigated for the presence of HPV, and the different genotypes. Results were evaluated based on the HPV positivity in different cytological findings, and ages. Furthermore, distribution of high-risk (HR) and low-risk (LR) genotypes in different groups was investigated. RESULTS: Among all participants, 1925 (78.1%) was with the normal cytological condition, 354 (14.4%) with ASC-US; 151 (6.1%) with low-grade squamous intraepithelial lesion (LSIL), and 34 (1.4%) with high-grade squamous intraepithelial lesion (HSIL). Our results showed that 649 out of 2464 patients (26.3%) were positive, and 1815 (73.7%) were negative for the presence of HPV. Among 649 positive patients, 223 (34.3%) were found positive for more than one genotype. HPV 16 was found the most common HR-HPV type in ASC-US and LSIL whereas HPV 18 was the most common in HSIL. HPV 6 was found the most common LR-HPV type in ASC-US and LSIL whereas HPV 11 was the most common in HSIL. 26.9% of women <50 years old, and 22.3% of women ≥50 years old was positive for HPV. The most common HR-HPV genotype was 16 in both groups with (19%) or without (17%) abnormal cytology. CONCLUSIONS: We concluded that HPV prevalence and genotype distribution in women with or without clinical findings is an important predictor of cervical cancer.

DHEA supplementation improves endometrial HOXA-10 mRNA expression in poor responders.

OBJECTIVE: The study was planned to investigate whether DHEA supplementation had an impact on endometrial receptivity in women who were poor responders (POR). MATERIAL AND METHODS: Twenty-eight POR women who were undergoing hysteroscopy and five fertile control subjects were included. The POR women were equally subdivided into two separate groups as patients who were currently using DHEA and those who were not. Endometrial samples of the subjects were obtained during hysteroscopy at the late follicular phase. Expression levels of endometrial HOXA-10, HOXA-11, and LIF mRNA were measured with the using real-time polymerase chain reaction. Spontaneous clinical pregnancy rates were also noted. RESULTS: Compared with POR women who were not given DHEA, upregulated endometrial HOXA-10 (7.33-fold) and HOXA-11 (2.39-fold) mRNA expression were detected in POR women on DHEA. The increase in HOXA-10 mRNA was significant (p<0.03). The fold increase in HOXA-11 mRNA was found as 2.39, which indicated a positive upregulation. However, this fold increment was insignificant (p<0.45). An insignificant increase in spontaneous clinical pregnancy rates in POR women on DHEA (53.3%) was observed compared with POR women who were not given DHEA (43.8%). CONCLUSION: Oral DHEA supplementation in POR upregulates endometrial HOXA-10 mRNA expression, which is known to positively modulate endometrial receptivity.

Evaluation on the Practice and Behaviour of Women Applied for Gynecology Outpatient Clinics About Screening Methods for Early Diagnosis of Breast Cancer.

OBJECTIVE: Breast self-examination (BSE), clinical breast exam (CBE), mammography and ultrasound imaging (UI) are screening methods used for early diagnosis of breast cancer (BC). The purpose of this study is to put forth the utilization frequency of these screening methods among women presenting to the gynecology outpatient clinics and the relation of these data with the socio-demographic characteristics of the women. MATERIALS AND METHODS: A survey was conducted among 429 women (age, 16-80 years) who were admitted to the gynecology outpatient clinics. The survey inquired about the rate and frequency of the performance of BSE, CBE, mammography and UI; personal and family history of breast cancer and social-demographic characteristics of the women. RESULTS: The mean age was 40.08 (SD: 3.67). More than half of the women above 40 years of age (59.7 %) had never undergone mammography. 99.8 % of the women who had undergone mammography had also received ultrasound imaging. A significant relationship was identified between the BSE performance and having mammography. 57.4% of the women above 40 years of age (117) had UI, 53.9% (110) had CBE and 57.3% (117) performed BSE. There was a significant relationship between the age, education status and regular BSE; positive family history of BC and having CBE and mammography. CONCLUSION: The results reveal that the rate of BSE performance, having mammography and CBE are at less-than-ideal levels. In this context, it is apparent that breast cancer screening methods are needed to be introduced and guidance about their application frequency should be provided for women in gynecology outpatient clinics.

Concomitant use of transvaginal sonography and Doppler indices improve diagnosis of adenomyosis.

This study was planned to investigate whether measuring of Doppler indices with TV-DUS improved the diagnosis of adenomyosis. Preoperative Doppler indices of subjects with a preliminary diagnosis of adenomyosis were compared with the histopathological results of excised specimens. Sensitivity, specificity and positive/negative predictive values (PPV, NPV) of the measured indices were also calculated. Sensitivity, specificity, PPV and NPV and positive and negative likelihood ratios (LR+) and (LR-) of TV-US in the diagnosis of adenomyosis were found to be 70.8%, 62.1%, 40.4%, 85.4%, 1.96 and 0.47, respectively. Sensitivity, specificity, PPV, NPV, LR + and LR - in the diagnosis of adenomyosis following the addition of TV-DUS were found to be 90%, 94.2%, 81.8%, 97%, 15.5 and 0.10, respectively. Concomitant use of TV-US and TV-DUS improved correct diagnosis of adenomyosis with high sensitivity.

Frozen embryo transfer prevents the detrimental effect of high estrogen on endometrium receptivity.

OBJECTIVE: To investigate whether serum levels of estradiol affect reproductive outcomes of normoresponder women undergoing fresh embryo transfer (ET) versus frozen-thawed ET (FET). MATERIAL AND METHODS: Two hundred fifty-five normoresponder women underwent fresh ET in their first or second in vitro fertilization cycle. Ninety-two women with negative pregnacy test results underwent FET. Clinical and ongoing pregnancy rates, implantation, and live birth rates of women undergoing fresh ET versus FET were compared. RESULTS: One hundred forty-seven (57.65%) out of the 255 normoresponder women receiving FET had positive beta-human chorionic gonadotrophin (hCG) results. The remaining 108 women had negative beta-hCG results. The clinical pregnancy rates of the fresh ET group were found as 55.69% (n=142). Ninety-two of the 108 women with failed pregnancies underwent FET; 72.83% had positive beta-hCG results (n=67), and 70.65% had clinical pregnancy (n=65). Both biochemical and clinical pregnancy rates of women undergoing FET increased significantly (p<0.012 and p<0.013, respectively). Ongoing pregnancy (60.87% vs. 52.94%) and live birth rates (59.87% vs. 48.63%) were similar in both fresh and FET groups. Serum E2 levels of women who failed to conceive were significantly higher than those women did conceive. Serum progesterone levels of women who conceived versus those that did not were similar. CONCLUSION: The detrimental effect of high serum estradiol levels on endometrial receptivity could be prevented by FET.

Follicular fluid cerebellin and betatrophin regulate the metabolic functions of growing follicles in polycystic ovary syndrome.

OBJECTIVE: The aim of this study was to assess the changes of follicular fluid (FF) and serum levels of cerebellin precursor protein 1 (cbln1) and betatrophin in patients with polycystic ovary syndrome (PCOS) undergoing in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) with a gonadotropin-releasing hormone (GnRH) antagonist protocol. METHODS: Twenty infertile women with PCOS and 20 control women diagnosed as poor responders undergoing ovarian stimulation with a GnRH antagonist were included. Blood samples were obtained during ovum pick-up. Follicular fluid from a dominant follicle was collected from the subjects. Using enzyme-linked immunosorbent assays, FF and serum levels of cbln1 and betatrophin were measured in both groups of participants. Metabolic and hormonal parameters were also determined and correlated with each other. RESULTS: Both groups of women had similar serum and FF betatrophin levels (55.0±8.9 ng/mL vs. 53.1±10.3 ng/mL, p=0.11). The serum and FF betatrophin levels of poor responders were found to be similar (49.9±5.9 ng/mL vs. 48.9±10.7 ng/mL, p=0.22). Conversely, the FF cbln1 levels of PCOS women were found to be significantly higher than the serum cbln1 levels (589.1±147.6 ng/L vs. 531.7±74.3 ng/L, p<0.02). The FF cbln1 levels of control participants without PCOS were significantly higher than their serum cbln1 levels (599.3±211.5 ng/L vs. 525.3±87.0 ng/L, p=0.01). Positive correlations were detected among body mass index, insulin resistance, serum insulin, total testosterone, and betatrophin levels in the PCOS group. CONCLUSION: Follicular fluid betatrophin and cbln1 concentrations may play a pivotal role on follicular growth in PCOS subjects undergoing IVF/ICSI with an antagonist protocol.

Impact of Endometrioma Resection on Eutopic Endometrium Metabolite Contents: Noninvasive Evaluation of Endometrium Receptivity.

The aim of this study was to determine whether endometrioma resection alters most commonly defined endometrial metabolites, lactate (Lac), N-acetylaspartate (NAA), creatine 1 (Cr1), creatine 2 (Cr2), and choline (Cho) during the window of implantation. Twenty patients with uni- or bilateral endometrioma and 7 patients having nonendometriotic benign ovarian cyst were included. Midluteal phase magnetic resonance spectroscopy analysis of eutopic endometrium was performed before surgery. Second spectrum of endometrium was obtained 3 to 5 months after laparoscopic endometrioma resection. Pre- and postoperative endometrial peaks of Lac, NAA, Cr, and Cho were measured in units and denominated in parts per million (ppm). Compared to preoperative peak values, significantly decreased NAA, Lac, and Cr1 signals were noted in patients undergoing endometrioma surgery. Nearly 5-fold decline in the NAA signal occurred after endometrioma surgery (1.94 ± 3.24 vs 0.37 ± 0.55). Likewise, 2.5-fold decline in Lac signals was noted after endometrioma resection (2.81 ± 2.64 vs 1.06 ± 1.88). Both uni- and bilateral endometrioma affected endometrium signals the same. The peak intensity of Cho, Cr1, Cr2, NAA, and Lac did not alter significantly after nonendometriotic cyst surgery. Endometrioma surgery straightens endometrial NAA, Lac, and Cr1 peaks, suggesting improvement in endometrial receptivity.

Uterine sarcoma in a 14 year-old girl presenting with uterine rupture.

•Uterine sarcomas are rare in adolescents. Adenosarcomas are even more rare. Uterine rupture, as a presentation is a rare entity.•It is hard to diagnose sarcomas preoperatively. It is still hard even postoperatively due to variable morphologic features.•Experience of the gynecopathologists and oncologic surgeons is the hallmark of the treatment option.