Prolidase activity in women with polycystic ovarian syndrome undergoing assisted conception.

Background: Prolidase is a manganese (Mn)-dependent cytosolic exopeptidase that degrades imidodipeptides with C-terminal proline or hydroxyproline. Prolidase recycling from imidodipeptides plays a critical role in collagen resynthesis and extracellular matrix (ECM) remodelling. Following an increase in gonadotropins, ovarian and follicular collagen undergo substantial degradation. Abnormal ovarian ECM composition is associated with polycystic ovary syndrome (PCOS). This study aimed to examine prolidase activity in the serum and follicular fluid (FF) of women undergoing in vitro fertilisation/intracytoplasmic sperm injection (IVF/ICSI) treatment, comparing those with PCOS to those with normal ovarian function.Methods: This prospective study enrolled 50 participants, of whom 44 were included. PCOS diagnosis followed the Rotterdam consensus criteria, with 20 patients constituting the study group. The control group comprised 24 individuals with mild-to-moderate male infertility. Prolidase enzyme activity in serum and FF was measured using the Chinard reagent via spectrophotometric analysis and compared between the groups.Results: Serum and FF prolidase levels were significantly lower in patients with PCOS (p < 0.05). A direct correlation was observed between serum and FF prolidase levels (p < 0.05). Although blastocyst quality scoring (BQS) significantly decreased in PCOS patients, no statistical difference was observed in the clinical pregnancy rate between the groups (p < 0.05) (p > 0.05). A negative correlation existed between serum prolidase levels and total antral follicle (AF) count (p < 0.05). Conversely, both serum and FF prolidase levels positively correlated with BQS (r = 0.574)(p < 0.05) (r = 0.650)(p < 0.05).Conclusions: Patients with PCOS showed lower serum and FF prolidase levels, indicating abnormal degradation of ovarian and follicular collagen, potentially causing anovulation.

Polycystic ovary syndrome (PCOS), the most prevalent endocrinopathy among reproductive-aged women, affects approximately 3­15% of this demographic. Long-term disorders such as cardiovascular disease, type 2 diabetes mellitus, obesity, and infertility are commonly associated with PCOS, with approximately 70% of affected women experiencing infertility. Although the aetiology of PCOS remains unclear, complex multigenic disorders and environmental factors such as abnormal ovarian extracellular matrix composition, disruption of the inflammatory pathway, and lifestyle factors have been found to be related.This study addresses the aetiology of PCOS, focusing on the close association between abnormal ovarian extracellular matrix composition and the syndrome, as seen in previous reports. Prolidase is a manganese-dependent cytosolic exopeptidase that degrades imidodipeptides using the C-terminal proline or hydroxyproline. Proline recycling from imidodipeptides by prolidase plays a critical role in the resynthesis of collagen and remodelling of the extracellular matrix. Our aim was to evaluate prolidase activity in the serum and follicular fluid of women diagnosed with PCOS. Our findings revealed a direct correlation between serum and follicular fluid prolidase levels, both of which were diminished in women with PCOS. Furthermore, a negative correlation was observed between serum prolidase levels and total antral follicle count indicating a potential link between prolidase activity and ovarian follicle development. In contrast, both serum and follicular fluid prolidase levels were positively correlated with blastocyst quality. In conclusion, PCOS patients showed lower serum and follicular fluid prolidase levels, indicating abnormal degradation of ovarian and follicular collagen, and potentially causing anovulation. Future studies measuring manganese levels in larger numbers of participants are required.

Comparative analysis of copper intrauterine device impact on female sexual dysfunction subtypes.

OBJECTIVES: To examine the effect of copper intrauterine device (Cu-IUD) on female sexual dysfunction (FSD) subtypes. MATERIAL AND METHODS: There were 159 sexually active women (ninety Cu-IUD users and sixty-nine women with no contraception) who attended the gynecology clinic for routine gynecologic control informed about the study and asked to fill Female Sexual Function Index (FSFI) and Beck Depression Inventory questionnaires. RESULTS: The prevalence of FSD was 41.1% (n=37) and 37.7% (n=26) in Cu-IUD users and control groups, respectively (p > 0.05). In analyses of mean overall and subgroup scores of FSFI, significantly lower scores for arousal (p=0.021), lubrication (p=0.021), orgasm (p=0.040), pain (p < 0.001), and overall FSFI (p=0.031) were noted in Cu-IUD users. When the results for FSFI domains were considered for Cu-IUD users separately, the only difference to reach statistical significance, using a Bonferroni adjustment, was found to be the pain domain. Finally, we determined that Cu-IUD status made the strongest unique contribution to explaining the dependent variable pain in multiple logistic regression model (ß = -0.26, p=0.001). CONCLUSION: Cu-IUD users have increased sexual pain compared to women with no contraception, which in turn possibly causes decreased sexual arousal, lubrication, and orgasm in these women.

Comparison of efficacy of bromocriptine and cabergoline to GnRH agonist in a rat endometriosis model.

OBJECTIVE: To determine the effect of dopamine agonists in a surgically induced endometriosis model on rats. STUDY DESIGN: In this prospective randomized experimental study, surgical induction of endometriosis was performed by autotransplantation technique on 52 adult female Wistar-Albino rats. Endometriosis formation was confirmed by a second-look laparotomy (n:48) 1 month later. Four study groups were randomly generated according to their treatment regimens: group 1 (leuprolide acetate, n = 12), group 2 (bromocriptine, n = 12), group 3 (cabergoline, n = 12) and group 4 (control, n = 12). Endometriotic implants were excised for histopathological examination after treatment at the setting of laparotomy. The mean surface areas and histopathological glandular tissue (GT) and stromal tissue (ST) scores of endometriotic implants were studied and compared among groups. RESULTS: After 30 days of treatment, the mean surface area of the endometriotic implants of leuprolide acetate, bromocriptine and cabergoline groups was significantly decreased. The regression of endometriotic foci size in comparison to control was highest in group 1, followed by group 2, then group 3. In the histopathological evaluation both the ST and GT scores of group 1, 2 and 3 were significantly decreased in comparison to controls without a statistically significant difference between the groups. CONCLUSION: Dopamine agonists are as effective as GnRH agonists in the regression of experimental endometriotic implants in rats. Further trials are needed to elucidate the pathways affected by dopamine agonists.

Effects of maternal ageing on ICSI outcomes and embryo development in relation to oocytes morphological characteristics of birefringent structures.

The aim of this study was to determine the morphological characteristics of the older reproductive aged women's oocytes and to reveal the influence of these characteristics on intra-cytoplasmic sperm injection (ICSI) outcomes. The oocytes of women older than 35 years of age were evaluated retrospectively. Non-invasive polarization microscopy (PolScope) examinations of mature oocytes were performed by measurement of meiotic spindles' length, area and retardance and zona pellucida thickness and retardance. Fertilization and conception competence and the correlation with the birefringent structures were assessed. Two hundred and thirteen mature oocytes from 54 women were evaluated with a PolScope. Length of the meiotic spindle was shown to be related to fertilization success of women with advanced maternal age. In conclusion, the PolScope is a useful device used to identify the oocyte quality. Quantitative measurements of meiotic spindle parameters may be valuable for the selection of high-quality oocytes that have the potential for embryo development in the in vitro fertilization (IVF) laboratory of women older than 35 years of age who are mostly poor responders.

Random-start controlled ovarian hyperstimulation with letrozole for fertility preservation in cancer patients: case series and review of literature.

The aim of this case series was to describe our experience with random-start controlled ovarian hyperstimulation (RS-COH) with the use of letrozole for fertility preservation. GnRH antagonist and letrozole cycles were started in three patients with a diagnosis of cancer and had a limited time window for fertility preservation for COH before initiating cancer therapy. Cycles were started in the late follicular or luteal phase, and the duration of COH ranged between 7-8 days. A total of 4-14 oocytes were retrieved, the peak E2 levels were 252-354 pg/ml and the saved time for start of the cancer treatment were 16-26 days for each patient. In conclusion, RS-COH with letrozole cycle is a reasonable option for fertility preservation in cancer patients for whom the treatment window may be narrow. Also, the use of a letrozole for COH may decrease the potential risk of ovarian hyperstimulation syndrome.

Pregnancy outcomes in a university hospital after legal requirement for single-embryo transfer.

OBJECTIVE: To present our in vitro fertilization outcomes after implementation of the Turkish government policy limiting the number of transferred embryos to one, in women under the age of 35, and two, in women over age 35; and to demonstrate the feasibility of this policy. STUDY DESIGN: We retrospectively reviewed the records of 423 patients who underwent ovarian stimulation, oocyte pick-up, intracytoplasmic sperm injection and embryo transfer due to primary or secondary infertility over a 22-month time interval. Clinical pregnancy was defined as a fetal heartbeat in the seventh gestational week. Embryo transfers were carried out on day 3 or 5. Descriptive statistics are given as the number of subjects and percentages. RESULTS: A total of 353 embryo transfers were performed: 261 (73.9%) were single-embryo transfer (244 (69.1%) elective and 17 (4.8%) non-elective) and 92 (26.1%) were double-embryo transfer. Of the 244 elective single-embryo transfers, 6.6% (n=16) were performed using frozen-thawed embryos. The average patient age was 29.6±4.5 years in the single-embryo group and 36.5±2.4 years in the double-embryo group. The cumulative pregnancy rates per oocyte pick-up were similar in both groups: 41% (n=107) in the single-embryo group and 43.4% (n=40) in the double-embryo group (p=0.678). The cumulative live birth rate of the single-embryo group (32.1%) was not statistically different from the double-embryo group (35.8%) (p=0.518). The twin pregnancy rate after single-embro transfer was significantly lower than with double-embryo transfer (2.8% (n=3) vs. 32.5% (n=13); p<0.001) and 62.5% of the twin pregnancies occurred in women 35 years or older who underwent double-embryo transfer. No significant difference in the spontaneous abortion rates was recorded between the single- and double-embryo transfer groups (16 (6.1%) vs. 6 (6.5%); p=0.894). CONCLUSION: Single-embryo transfer results in a pregnancy rate comparable to double-embryo transfer, with a significantly reduced multiple pregnancy rate. Double-embryo transfer in patients over age 35 should be reconsidered because of the resulting high rate of multiple pregnancy.

Do quantitative birefringence characteristics of meiotic spindle and zona pellucida have an impact on implantation in single embryo transfer cycles?

OBJECTIVE: The aim of this study was to determine whether quantitative PolScope characteristics of meiotic spindle and zona pellucida could be used as a non-invasive marker to predict implantation success in elective single embryo transfer cycles. METHODS: Quantitative birefringence parameters; including mean retardance, area, length and polar body deviation angle of meiotic spindle and mean retardance and width of inner zona pellucida belonging to 53 transfer oocytes from elective single embryo transfer cycles were retrospectively analyzed. The relevant PolScope features were compared between 20 conception and 33 non-conception cycles. RESULTS: Meiotic spindle mean retardance, area, length and inner zona pellucida mean retardance and width did not reveal a statistically significant difference between transfer oocytes from conception and non-conception cycles. Deviation angle of the polar bodies was also comparable between the groups. Spindle and inner zona PolScope characteristics of transfer oocytes were not correlated with the maternal age. CONCLUSION: Quantitative PolScope features of meiotic spindle and inner zona pellucida can not be used as a non-invasive marker to predict assisted reproductive technology success in elective single embryo transfer cycles.

What should be the protocol selection after failure of in-vitro fertilization at normoresponder patients: Agonist or antagonist?

OBJECTIVE: Evaluation of the impact of agonist or antagonist protocol selection on pregnancy outcomes after failure of in-vitro fertilization (IVF) treatment cycles which were down regulated with Gonadotropin Releasing Hormone (GnRH) agonist. MATERIALS AND METHODS: This was a retrospective study. Two hundred and sixty nine patients who were treated with GnRH agonist protocol between years 2002-2012 at an IVF unit and underwent a second attempt following one year period after failure of IVF enrolled in the study. Age, basal FSH levels, antral follicle counts, duration of induction, the number of yielded oocytes, the number of transferred embryos and the transfer days, clinical and ongoing pregnancy rates were evaluated for each treatment cycle. RESULTS: Normoresponder patients were separated into two groups according to the agonist or antagonist protocol selection at the second attempt and the results of two consequent IVF cycles were compared. There were no statistically significant difference between the groups for the dosage of administered gonadotropin, duration of induction, the count of yielded oocytes, the day and the number of transferred embryos (p>0.05). Furthermore the fertilization rate, clinical and ongoing pregnancy rates were similar in two groups. CONCLUSION: The selection of antagonist treatment is effective as agonist protocols at normoresponder patients after failure of IVF.

Paracrine activation of WNT/ß-catenin pathway in uterine leiomyoma stem cells promotes tumor growth.

Uterine leiomyomas are extremely common estrogen and progesterone-dependent tumors of the myometrium and cause irregular uterine bleeding, severe anemia, and recurrent pregnancy loss in 15-30% of reproductive-age women. Each leiomyoma is thought to arise from a single mutated myometrial smooth muscle stem cell. Leiomyoma side-population (LMSP) cells comprising 1% of all tumor cells and displaying tumor-initiating stem cell characteristics are essential for estrogen- and progesterone-dependent in vivo growth of tumors, although they have remarkably lower estrogen/progesterone receptor levels than mature myometrial or leiomyoma cells. However, how estrogen/progesterone regulates the growth of LMSP cells via mature neighboring cells is unknown. Here, we demonstrate a critical paracrine role of the wingless-type (WNT)/ß-catenin pathway in estrogen/progesterone-dependent tumorigenesis, involving LMSP and differentiated myometrial or leiomyoma cells. Estrogen/progesterone treatment of mature myometrial cells induced expression of WNT11 and WNT16, which remained constitutively elevated in leiomyoma tissues. In LMSP cells cocultured with mature myometrial cells, estrogen-progesterone selectively induced nuclear translocation of ß-catenin and induced transcriptional activity of its heterodimeric partner T-cell factor and their target gene AXIN2, leading to the proliferation of LMSP cells. This effect could be blocked by a WNT antagonist. Ectopic expression of inhibitor of ß-catenin and T-cell factor 4 in LMSP cells, but not in mature leiomyoma cells, blocked the estrogen/progesterone-dependent growth of human tumors in vivo. We uncovered a paracrine role of the WNT/ß-catenin pathway that enables mature myometrial or leiomyoma cells to send mitogenic signals to neighboring tissue stem cells in response to estrogen and progesterone, leading to the growth of uterine leiomyomas.

Effects of smoking during pregnancy on DNA damage and ROS level consequences in maternal and newborns' blood.

Some of the genotoxic/carcinogenic substances or metabolites in cigarette smoke are capable of passing through the placenta and harming a newborn's health. Smoking is also known as a factor in the formation of oxidative damage and the main mechanism involved in the carcinogenic process. Predetermining this genotoxic risk can be successfully achieved by measuring certain parameters of oxidative stress. The comet assay is considered an important biomarker for the evaluation of genotoxic substances and is effective for detecting DNA damage caused by smoking. This study examined third trimester bloods and the cord blood of 28 actively smoking and 22 non-smoking mothers in terms of DNA damage and oxidative stress parameters. Cu/Zn superoxide dismutase (CuZn-SOD), malondialdehyde (MDA), catalase (CAT), plasma nitrite/nitrates (NO2-/NO3-), selenium-dependent glutathione peroxidase (Se-GPx), Cu, and Zn levels were measured as indicators of oxidative damage. There were no significant increases in DNA damage of the actively smoking pregnant group in comparison with the non-smoking pregnant group, either in the third trimester or cord blood. Oxidative stress parameters of smoker and non-smoker groups were statistically different for MDA (p<0.05), CuZn-SOD (p<0.01), Se-GPx (p<0.05) values while the difference was not significant for NO2-/NO3-, CAT, Zn, and Cu values. The same values were also investigated in cord blood,and only NO2/NO3-(p<0.01), Se-GPx (p<0.01 and CAT (p<0.001) values were found statistically different.Smoking mothers may have been exposed to more oxidative stress than non-smoking mothers

Effect of a low dose of gonadotropin releasing hormone antagonist on in vitro fertilization outcomes.

PURPOSE: This study aims to determine whether a low dose of the gonadotropin releasing hormone antagonist, cetrorelix, prevents a premature luteinizing hormone (LH) surge and affects in vitro fertilization (IVF) outcomes compared to the standard dose of 0.25 mg/day. METHODS: In this study, 45 IVF/intracytoplasmic sperm injection patients were stimulated with recombinant follicle stimulation hormone from day 2 of the cycle. Cetrorelix was injected daily from day 6 of gonadotropin administration. Twenty-two patients received cetrorelix at a dose of 0.25 mg/day, whereas 23 participants received half dose. RESULTS: The mean consumption of gonadotropins was significantly higher in patients receiving 0.25 mg/day of cetrorelix (2,213 vs. 1,350 U; p = 0.046). The clinical pregnancy rates were similar in both groups (31.8 vs. 47.8 %; p = 0.273). Premature LH surge was detected in 9.1 % of the patients receiving cetrorelix 0.25 mg/day and in 13 % of the patients receiving cetrorelix 0.125 mg/day (p > 0.05). The difference between two groups was not statistically significant (p > 0.05). CONCLUSIONS: Our results suggest that there is no difference between a cetrorelix dose of 0.125 or 0.25 mg/day in preventing premature LH rise during ovarian stimulation for IVF.

Ovarian reserve testing before and after laparoscopic tubal bipolar electrodesiccation and transection.

OBJECTIVE: To find out whether tubal sterilization leads to loss of ovarian reserve, we assessed the hormonal and ultrasonographic parameters of ovarian reserve in women who underwent laparoscopic tubal sterilization by bipolar electrodesiccation and transection. STUDY DESIGN: In this preliminary study, laparoscopic tubal sterilization was performed on 49 healthy women who had voluntarily requested elective surgical sterilization. Among the current ovarian reserve indicators, in the early proliferative phases, preoperative (baseline) and postoperative (third month) serum follicle-stimulating hormone (FSH), estradiol (E2), and anti-Mullerian hormone (AMH) levels, ovarian volume, and antral follicle counts (AFCs) were determined. Analysis of these hormonal and ultrasonographic parameters of ovarian reserve preoperatively and postoperatively was the main outcome measure. RESULTS: Preoperative and third-month postoperative FSH, LH, E2, and AMH levels did not reveal statistically significant differences (p=0.101, p=0.180, p=0.254, and p=0.079; respectively). The ultrasonographic indicators of ovarian reserve did not change in terms of total ovarian volume and total AFC (p=0.793 and p=0.098, respectively). CONCLUSIONS: Short-term follow-up study results revealed a slight but non-significant change in the current ovarian reserve markers, especially in the AMH levels.

A macrophage activation marker chitotriosidase in women with PCOS: does low-grade chronic inflammation in PCOS relate to PCOS itself or obesity?

OBJECTIVE: Polycystic ovary syndrome (PCOS) is described as a low-grade chronic inflammatory state where the macrophage infiltration increased in visceral fat play the major role. The aim of this study was to assess chitotriosidase (ChT) activity (an activated macrophage marker) in women with PCOS and controls and to evaluate its correlation with anthropometric and biochemical parameters. STUDY DESIGN: Seventy-nine women with PCOS and 60 healthy controls were included in the study. PCOS and controls were divided into two subgroups according to body mass indexes (BMIs) as normoweight (<25 kg/m(2)) and overweight (≥25 kg/m(2)). ChT activity, biochemical (free testosterone, luteinizing hormone, insulin resistance index, etc.) and clinical parameters [BMI, waist-to-hip ratio, modified Ferriman-Gallwey scores (mFG)] were analyzed according to groups. RESULTS: Serum ChT activity was significantly (p < 0.001) higher in women with PCOS than controls (normoweight, 87.1 ± 90.1 vs. 18.4 ± 9.0 nmol/ml/h; overweight, 92.0 ± 96.7 vs. 17.9 ± 12.1 nmol/ml/h PCOS and controls, respectively). No statistically significant difference was noted between ChT activity of normoweight and overweight PCOS subgroups. A positive correlation was found between ChT activity and PCOS status (r 0.598, p < 0.001), mFG scores (r 0.525, p < 0.001), free testosterone (r 0.402, p 0.001) and total testosterone (r 0.168, p 0.048) for the combined groups (PCOS + controls). In multivariant linear regression analysis participants' PCOS status (presence or absence) and LH levels were the main predictors of ChT activity in the whole study population (p 0.002 and p 0.041, respectively). CONCLUSION: ChT activity elevates in PCOS independent of obesity. Our findings support the concept of PCOS is a state of low-grade chronic inflammation where the macrophages could play the major role. Hyperandrogenism might also be related to this inflammatory state and can be a subject of further trials.

Combined approach in a large placental chorioangioma case with intratumoral alcohol injection, cordocentesis, IU transfusion, and amnioreduction.

Large placental chorioangiomas (LPCs) are rare complications of pregnancy and these tumors are associated with increased risk of pregnancy complications. We presented a LPC case at 25th week of gestation, complicated by polyhydramnios and preterm labor in which intratumoral alcohol injection was performed succesfully. Cordocentesis, fetal intrauterine transfusion, and amnioreduction were performed at the same session, as a combined approach. The safety of the procedure was shown by the lack of alcohol in the fetal cord blood sample. Intratumoral alcohol injection is a reliable and effective treatment modality in the management of LPCs and may be combined with other invasive procedures.

Chitotriosidase, interleukin-1 beta and tumor necrosis factor alpha levels in mild preeclampsia.

OBJECTIVE: Chitotriosidase (ChT) is an activated macrophage marker. Tumor necrosis factor alpha (TNF-α) and interleukin 1 beta (IL-1ß) are mainly produced macrophages. The aim of the present study was to evaluate the relationship between serum ChT activity, levels of TNF-α and IL-1ß in patients with mild preeclampsia and normal pregnancy. METHODS: An overall 64 cases, 32 healthy pregnant control women (control group) and 32 women with mild preeclamptic patients (study group), were enrolled in this study. At the beginning of the study, all study participants were matched for age and gestational age. Serum ChT activity was measured by fluorometer; TNF-α and IL-1ß levels were measured by enzyme-linked immunosorbent assay. RESULTS: The mean age, gestational week, parity and gravida were similar in the two groups (p > 0.05). Serum ChT activity was significantly higher in the preeclampsia group compared to the control group (p < 0.05). Levels of TNF-α and IL-1ß in patients with mild preeclampsia were similar compared to the control group (p > 0.05). In the PE group, serum ChT activity was not correlated with TNF-α and IL-1ß. CONCLUSION: Mild preeclampsia is found associated with higher ChT activity. This result suggests that activated macrophages play a role in the pathogenesis of preeclampsia. This suggestion needs to be confirmed in future studies with larger populations.

Chitotriosidase levels in patients with severe endometriosis.

OBJECTIVE: To study the levels of chitotriosidase activity in the peritoneal fluid and the plasma of patients with severe endometriosis and control subjects. MATERIALS AND METHODS: Twenty-five women with laparoscopically and histopathologically confirmed endometriosis (study group) and 27 control patients who had undergone laparoscopic surgery were included. Peritoneal fluid and peripheral blood were obtained from all the patients before the surgery. Chitotriosidase activities were measured. RESULTS: Analysis of chitotriosidase activity in the peritoneal fluid of patients with endometriosis showed that there was no significant difference between endometriosis and control group, respectively (32.04 ± 64.20 vs. 15.25 ± 31.17 nmol/mL/h; p > 0.05). Analysis of chitotriosidase activity in plasma of patients with endometriosis showed significantly increased levels of chitotriosidase levels compared with the control group (74.81 ± 60.54 vs. 14.10 ± 26.17; p < 0.001), respectively. CONCLUSION: We found that the activity of chitotriosidase in plasma was statistically higher in severe endometriosis patients than women without endometriosis.

Toluidine blue staining in the diagnosis of endometrial pathologies: a preliminary study before chromohysteroscopy.

OBJECTIVE: To assess the efficacy of toluidine blue (TBlue) staining in the detection of endometrial premalignant or malignant lesions in uterine specimens obtained from hysterectomies. We hoped that the results of this preliminary study would help us in our future studies which may be on chromohysteroscopy. STUDY DESIGN: We developed an endometrium staining technique in which TBlue solution was used as a vital dye. The study enrolled 50 patients. Of these, 20 patients known to have a premalign or malign lesion of the uterus constituted the study group and 30 patients were recruited as the control group who were operated for leiomyoma of the uterus. Immediately after hysterectomy, all specimens were dyed with TBlue solution and a frozen section procedure was performed on the positively stained areas. The pre- and post-operative histopathological diagnoses of the participants were compared and the reliability of the new method was calculated. RESULTS: The sensitivity of TBlue staining in the determination of endometrial pathologies was 100% (95%, CI 0.83-1) with a specificity of 90% (95%, CI 0.74-0.96). As a diagnostic test, the positive predictive value (PPV) was 87% with a negative predictive value (NPV) of 100%. The positive likelihood ratio (+LR) was 10 (95%, CI 3.41-29.2). CONCLUSIONS: Our preliminary results demonstrated that TBlue staining of endometrium is a reliable and highly sensitive technique that may be used in the frozen section examination of uterine specimens if a malignancy is suspected. Moreover, TBlue staining of endometrium may prove useful in hysteroscopy, although the technique, interpretation, and indications require further studies.