Ovulation stigma and concentration of progesterone and estradiol in peritoneal fluid: relation with fertility and endometriosis

The relationship between the presence or absence of an ovulation stigma and (1) the fertility status, (2) the incidence of endometriosis, (3) the concentration of progesterone and estradiol in the peritoneal fluid, and (4) the blood levels of luteinizing hormone, follicle-stimulating hormone, progesterone, and estradiol in 21 fertile and 45 infertile patients who underwent a laparoscopy in the early (n = 48) or late luteal phase (n = 18) was investigated. An ovulation stigma was observed in about half of the patients, irrespective of their fertility status (past and subsequent), the presence of endometriosis, or the time of the luteal phase. Progesterone and estradiol concentrations in the peritoneal fluid were highest in the early luteal phase, but they were not correlated with the presence or absence of an ovulation stigma. No significant differences were observed in peripheral hormone levels between women with and those without an ovulation stigma nor between women with high or low concentrations of progesterone in the peritoneal fluid. From the data, it is concluded that hormone assays are of no aid in the diagnosis of the luteinized unruptured follicle syndrome and that the absence of an ovulation stigma on laparoscopic examination cannot be equated with the luteinized unruptured follicle syndrome.     

Clinical significance of the luteinized unruptured follicle syndrome as a cause of infertility

The luteinized unruptured follicle syndrome (LUF) exists and is a cause of infertility. Although the data of the laparoscopic inspection of the ovaries and the corpora lutea and of steroid hormone concentrations in peritoneal fluid constitute strong evidence that the syndrome exists, its role as a cause of infertility is less clear. The only evidence available that the LUF syndrome is a cause of infertility, is the finding that the syndrome occurs statistically more frequently in women with unexplained infertility than in a control group of women. It still has to be proven whether the LUF syndrome occurs repetitively in each cycle and causes infertility, or whether the syndrome occurs intermittently and only reduces the probability of conception.Diagnosis of the syndrome can be made by laparoscopic inspection of the ovaries and by the assay of 17β-estradiol and progesterone, in peritoneal fluid between day 14 and 20 of the cycle.The relationship between the LUF syndrome and pelvic endometriosis and luteal phase insufficiency is discussed. We suggest that the LUF syndrome might be the cause of endometriosis thus explaining the statistical association between both syndromes, and the infertility of women with only mild endometriosis.We favor the hypothesis that the LUF syndrome might be caused by stress thus constituting a ‘psychological infertility’ and we suggest that the syndrome could explain the spontaneous cure rate. Therefore, before any therapy can be accepted as the treatment of the LUF syndrome, it should be strictly assessed with adequate controls.     

Incidence, recurrence and treatment of the luteinized unruptured follicle syndrome

The frequency of the luteinized unruptured follicle (LUF) was determined in a population of 220 regularly cycling women, infertile for at least two years. Laparoscopy was performed during the very first days of the luteal phase. In 26 women without other demonstrable cause of infertility, a diagnosis of LUF was made based on the absence of an ovulation stigma and the low concentrations of progesterone (P) and 17 beta-oestradiol (E2) in peritoneal fluid (PF). Twenty of these 26 women underwent a culdocentesis 72-96 hours after the serum LH-rise in a following cycle. In 19 out of 20, low P and E2 concentrations in PF were again found, suggesting the recurrence of LUF. Subsequently, ovulation was induced with human menopausal gonadotrophins (hMG) alone (n = 4), or in combination with human chorionic gonadotrophin (hCG, n = 9). At carefully timed culdocentesis (at LH/hCG + 72-96 hours), P concentration in PF was high in the hMG-hCG treated women but remained low in those given hMG alone. The combination of hMG and hCG may be a valuable treatment of LUF.     

Peritoneal fluid in female fertility and sterility (author's transl)

The authors have analyzed samples of peritoneal liquid to determine how and in which measure the level of steroid hormones allows to distinguish between follicular rupture and ovulation, and follicular luteinization without rupture and lack of ovulation. Volume of peritoneal fluid is not influenced by endometriosis or by pelvic varicosities, but it increases during the luteal phase; peritoneal protein concentration is also at its highest during the luteal phase. Progestin and estradiol-17 beta are higher in peritoneal fluid than in serum; such high concentration is maintained for at least a week after ovulation. The concentration of such hormones is higher in women with ovulatory scars than in those with luteinized unruptured follicle syndrome. Such findings show that peritoneal liquid is a transudate of the hyperemic active ovary, and that the level of concentration of progestin and estradiol-17 beta can be used in the diagnosis of luteinized unruptured follicle syndrome.     

Peritoneal fluid volume and steroid hormone concentrations in baboons with and without either spontaneous minimal/mild endometriosis or the luteinized unruptured follicle syndrome

This study was performed to document for the first time peritoneal fluid volumes and steroid hormone levels in baboons with and without endometriosis. A laparoscopy was carried out in 19 female baboons (11 with a normal pelvis and 8 with histologically proven spontaneous minimal/mild endometriosis) during 64 cycles in the early luteal phase. Peritoneal fluid was measured and aspirated. The pelvis was examined for evidence of recent ovulation and endometriosis. Peritoneal fluid and serum were assayed for 17beta-estradiol and progesterone. The mean volume of peritoneal fluid and its concentration of 17beta-estradiol and progesterone were 2.6 ml, 679 pmol/l and 64 nmol/1, respectively. No differences were observed between animals with or without either endometriosis or luteinized unruptured follicle syndrome. The peritoneal fluid and serum steroid levels were comparable to those found in women. The results presented are similar to those obtained in women with endometriosis and this suggests that the baboon can be used for the study of this disease.Supported by the Commission of the European Communities (DG VIII Development and DG XII Science, Research and Development) and by the Vlaamse Interuniversitaire Raad (Flemish Interuniversity Council), Brussels, Belgium     

Peritoneal fluid and serum steroids in infertility patients

Peritoneal fluid and serum were collected from 78 patients at the time of laparoscopy. Twenty-two were fertile controls (CTL), and 56 were infertility patients, who were subdivided into three main groups: endometriosis (EMS), pelvic adhesions (ADH), and ovarian dysfunction (OvDF). Based on control group data, biochemical criteria indicative of the presence of a stigma, S(+), were established: (1) serum progesterone (P) greater than or equal to 2 ng/ml, (2) peritoneal fluid P greater than or equal to 50 ng/ml, and (3) peritoneal fluid/serum ratio of P greater than or equal to 3. Direct visualization by laparoscopy showed that 21% CTL, 75% EMS, 69% ADH, and 56% OvDF subjects had luteinized unruptured follicle (LUF) syndrome. Biochemical criteria, however, demonstrated only 7% CTL, 37% EMS, 23% ADH, and 56% OvDF subjects had LUF. Peritoneal fluid estradiol (E2) and P concentrations and total content were significantly lower in LUF than in non-LUF patients, whereas serum E2 and P concentrations were not different between the two groups. Values for testosterone and androstenedione in peritoneal fluid and serum were similar between these two groups. Endometrial dating in LUF versus non-LUF patients were also similar. The usual indicators of ovulation, i.e., serum P, endometrial dating, and basal body temperature, failed to identify LUF. The diagnosis of LUF can be best made by P assay of peritoneal fluid and serum.     

Ovarian adhesions impair ovulation

The concentration of progesterone in peritoneal fluid, aspirated from the pouch of Douglas by laparoscopy, 72 hours after the peak of the luteinizing hormone in serum is believed to indicate whether the ovarian follicle has ruptured or not. Twenty six patients were studied. The volume of peritoneal fluid 72 hours after the peak of the luteinizing hormone was markedly decreased when bilateral ovarian adhesions were present. Progesterone concentrations, assayed during the early luteal phase in peritoneal fluid of women with bilateral ovarian adhesions, were significantly lower than in women with a corpus luteum presenting an ovulation stigma and even significantly lower than in those without ovulation stigma (luteinized unruptured follicle syndrome). The assay of progesterone in peritoneal fluid during the early luteal phase may be of value in women with ovarian adhesions.     

Incidence of the luteinized unruptured follicle syndrome in fertile women and in women with endometriosis

One-hundred normal fertile women with normal luteal phase and 118 women with endometriosis underwent luteal phase laparoscopy before day 22. The luteal phase was ascertained by the presence of secretory endometrium and serum progesterone levels higher than 3 ng/ml. The ovaries were carefully inspected for the presence or absence of an ovulation ostium. The percentage of ostii that was observed in fertile women (91%) was similar to that observed in women with mild endometriosis (85%). However, in women with moderate and severe endometriosis, significantly less ostii were noted, respectively 72 and 51%. It is therefore argued that the absence of an ovulation ostium (so-called luteinized unrupted follicle syndrome, LUF) is more frequent in women with moderate and severe endometriosis and may contribute to infertility in this group of women.     

Effect of ovarian involvement on the frequency of luteinized unruptured follicle in endometriosis.

OBJECTIVE: When we review the current literature on endometriosis and luteinized unruptured follicle (LUF), we see that most of the studies deal with only the association between LUF frequency and the severity of endometriosis. Our purpose was to evaluate the effect of ovarian involvement on LUF frequency in endometriosis and assess the relationship between endometriosis and LUF in infertile women. STUDY DESIGN: This study is a prospective analysis covering a total of 126 infertile women between 22 and 35 years of age who underwent diagnostic laparoscopy from September 1995 to August 1997 in the Department of Obstetrics and Gynecology at Süleyman Demirel University, Isparta, Turkey. Endometriosis was diagnosed in 58 of these patients. All had received a revised American Fertility Society staging score at the time of the laparoscopic diagnosis. Diagnosis of LUF was made when the following criteria were fulfilled: absence of ultrasonic signs of ovulation, and absence of an ovulation ostium on the follicle by laparoscopy, despite increased serum progesterone. Statistical evaluation was performed using chi2 test and Fisher's exact test where appropriate. RESULTS: The prevalence of LUF in mild, moderate and severe endometriosis cases was 13.3, 41.2 and 72.7%, respectively. The LUF frequencies were 45.9% in 37 endometriosis patients with ovarian involvement, 9.5% in 21 cases without ovarian involvement, and 5.9% in 68 cases without endometriosis. A statistically significant difference was observed between the LUF rate in the group with ovarian involvement and that without involvement (chi(2) = 8.06, p < 0.001). CONCLUSION: In summary, in this study we noted a significant increase in LUF frequency in endometriosis patients with ovarian involvement.     

Exaggerated prolactin response to thyrotropin-releasing hormone in infertile women with the luteinized unruptured follicle syndrome

Summary Ten cases of luteinized unruptured follicle (LUF) syndrome out of 250 women with unexplained infertility were detected on ultrasonography, giving a frequency of 4%. Hormonal analysis revealed lower serum progesterone levels at mid-luteal phase in LUF cases, suggesting a link between LUF syndrome and inadequate luteal phase. Prolactin response to thyrotropin-releasing hormone was exaggerated in LUF cases as compared with ovulatory cases. Aberrant prolactin release may be a contributory factor in the pathophysiology of the LUF syndrome.     

Oocyte retention after follicle luteinization

Indirect evidence supports the existence of the luteinized unruptured follicle syndrome in infertile women. To seek direct evidence of oocyte retention, infertile and normal women were studied in the early and midluteal phase by visual documentation of ovulation stigma, needle aspiration of ovarian follicles, and peritoneal fluid collection for estradiol and progesterone assay. Luteal phase was confirmed by endometrial biopsy (postovulation day 2 to 8). In normal control subjects (n = 16), 25% of test cycles were stigma-negative and no oocytes were recovered. In infertile group (n = 23), 43% of test cycles were stigma-negative. Five oocytes were recovered including one from a stigma-bearing follicle. Peritoneal fluid steroid levels failed to discriminate stigma-positive from stigma-negative cycles in either group. Oocyte retention after luteinization occurs in infertile women.     

Diagnosis of luteinized unruptured follicle (LUF) syndrome by ultrasound

To evaluate the role of ultrasound in diagnosing luteinized unruptured follicle (LUF), 37 women with unexplained infertility were examined for two to three menstrual cycles. Laparoscopy or laparotomy was performed on days 16 to 18 of the third study cycle in 25 patients. The LUF syndrome was suspected at ultrasound examination in 57 of 100 cycles observed. In the remaining 43 cycles, follicular collapse was observed in 33, and 10 were diagnosed as anovulatory. At laparoscopy or laparotomy on 25 patients, 18 of the 21 patients diagnosed as having LUF by ultrasound had a corpus luteum without a stigma. The other three cases diagnosed as LUF by ultrasound had ovulation stigmata. Additional findings in the 25 patients who underwent laparotomy or laparoscopy were endometriosis in 7 (5 of whom had LUF as well), ovulation in 5, bilateral hydrosalpinx in 1, and inability to visualize the ovaries because of adhesions in 1. The LUF syndrome was not a consistent change in the ovulatory pattern of most of the patients. It occurred by ultrasound diagnosis in three consecutive cycles in only 34% of patients.     

Association of interleukin-6 and estradiol with hepatocyte growth factor in peritoneal fluid of women with endometriosis

BACKGROUND: Different cytokines and ovarian steroid hormones have been reported to regulate the growth and maintenance of endometriosis. We determined the relationship between peritoneal fluid concentrations of interleukin-6, ovarian steroids and hepatocyte growth factor in different revised American Fertility Society (AFS) staging and morphologic appearances of endometriosis. METHODS: Peritoneal fluid was collected from 30 women with endometriosis and 20 women without endometriosis during laparoscopy, and hepatocyte growth factor, interleukin(IL)-6 and ovarian steroids were measured in peritoneal fluid. The concentrations of hepatocyte growth factor and IL-6 in peritoneal fluid were measured by ELISA, and that of estradiol and progesterone by using the immulyze-enzyme amplified luminescence system. Changes in peritoneal fluid concentrations of hepatocyte growth factor, IL-6, estradiol and progesterone in different stages and morphologic appearances of endometriosis were examined to demonstrate their differences in early and advanced endometriosis. RESULTS: Peritoneal fluid levels of hepatocyte growth factor in women with stage I-II endometriosis were significantly higher than in both women with stage III-IV endometriosis and without endometriosis. A similar significant increase in stage I-II endometriosis was also observed for IL-6 and estradiol. When we divided the women according to different morphologic appearances of endometriosis, we found significantly higher concentrations of hepatocyte growth factor (HGF), IL-6, estradiol and progesterone in women containing red lesions compared with other pigments or without endometriosis. A positive correlation was observed between peritoneal fluid levels of IL-6 and hepatocyte growth factor only but not between other markers. Although estradiol levels in peritoneal fluid showed an increased tendency to elevate in the proliferative phase of endometriosis women, hepatocyte growth factor and progesterone displayed higher concentrations in the secretory phase of the menstrual cycle. After adjusting different variables in peritoneal fluid, multiple analysis of covariance indicated that hepatocyte growth factor levels in peritoneal fluid were independently involved in the red morphologic activity of endometriosis. CONCLUSIONS: Early staging and red color appearance of endometriosis are associated with the elevation in peritoneal fluid concentrations of hepatocyte growth factor, IL-6 and estradiol, demonstrating the combined effect of these cytokines and ovarian steroids in the production of hepatocyte growth factor from endometrial tissues in active endometriosis.     

Follicle growth curves and hormonal patterns in patients with the luteinized unruptured follicle syndrome

A prospective longitudinal and standardized study is presented, dealing with ultrasonographic and hormonal characteristics of the luteinized unruptured follicle (LUF) syndrome. Among 600 cycles monitored in 270 infertility patients, 40 cycles in 27 patients showed no evidence of follicle rupture, in spite of signs of luteinization, as reflected by basal body temperature recordings and progesterone determinations. In this study, 20 LUF cycles in 20 infertile patients were compared with 45 ovulatory cycles in 45 control women. During the follicular phase, no substantial difference in follicle growth was found, but after the luteinizing hormone peak, LUF follicles, instead of rupturing, showed a typical accelerated growth pattern. Both mean luteinizing hormone peak levels and midluteal progesterone levels were significantly lower in LUF cycles than in the control cycles. However, the duration of the luteal phase was not affected. Both central and local factors can be held responsible for the lack of follicle rupture. Ultrasound offers new possibilities as a noninvasive method in diagnosing the LUF syndrome.     

Mild endometriosis and ovulatory dysfunction: effect of danazol treatment on success of ovulation induction

The effectiveness of ovulation induction with clomiphene citrate or human menopausal gonadotropins was evaluated in 52 infertile women with stage I or stage II endometriosis and ovulatory dysfunction: anovulation or luteinized unruptured follicle (LUF) syndrome before (group I) and after (group II) danazol treatment. The incidence of anovulation and LUF in the endometriosis population was 9% and 34%, respectively. In group I, 10 of 36 patients (27.8%) conceived, with an average of 17.6 induction cycles per pregnancy. In group II, 21 of 30 patients (70%) conceived, with an average of 4.5 cycles per pregnancy (difference significant at P less than 0.001). There was no difference in the average number of ovulation induction cycles per patient between groups I and II (4.9 and 3.1, respectively). Of 14 patients who did not conceive in group I and crossed over to group II, 9 (64.3%) conceived (not different from group II). Spontaneous abortion rates were 20% in group I and 14% in group II. These results indicate that mild endometriosis may interfere with conception through mechanisms other than ovulatory dysfunction and that treatment with danazol appears to more than double the fertility rate.     

LUF-Syndrom

The luteinized unruptured follicle (LUF) syndrome is estimated to be present in 6–12% of cases of female subfertility. When ovarian stimulation is used the incidence rises to 20–25%. A LUF syndrome tends to return in subsequent cycles. Diagnosis is made by several ultrasound scans starting the day before the expected ovulation. Typical for a LUF syndrome is the missing follicle collapse. There may be different causes leading to a LUF syndrome. The data in this review lead us to the conclusion that a LUF syndrome really exists. In fertile women the prevalence is about 10%, which will not reduce the fertility potential. One should keep in mind the possibility of LUF syndrome in idiopathic infertility. The only possible therapy is IVF as long as no other treatment options are available, which also are addressed in this review.     

Luteinized unruptured follicle syndrome in mild endometriosis. Assessment with biochemical parameters

Failure to extrude an ovum, with subsequent luteinization of the unruptured follicle (LUF), has been proposed as a cause of infertility in women with mild endometriosis. To assess the incidence of this process we performed laparoscopies in the early luteal phase on 16 women with mild endometriosis and 8 control subjects. Peritoneal fluid was aspirated and a plasma sample obtained concurrently. Estradiol (E2) and progesterone (P) concentrations were determined. A review of the literature suggested that the following hormonal criteria correlated with follicular rupture: fluid E2 greater than or equal to 500 pg/ml, E2 fluid/plasma ratio greater than or equal to 3.1, fluid P greater than or equal to 3,000 ng/dl and P fluid/plasma ratio greater than or equal to 5:1. All control subjects met at least one E2 and one P criterion: 75% met all. In contrast, less than one-third with mild endometriosis met all, and three (19%) met none. Five met only E2 criteria. These findings suggest that LUF occurs occasionally in association with mild endometriosis. Additionally, ovarian steroidogenesis, particularly P secretion, was impaired frequently in the absence of LUF in women with endometriosis.     

The origin of estrogens, progesterone, androgens and sex hormone binding globulin in peritoneal fluid in the immediate postovulatory period in normal ovulating women.

In 20 women with proven fertility, one menstrual cycle was monitored by ovarian ultrasonography, laparoscopy and estimation of 17 beta-estradiol (E2), estrone (E1), progesterone, testosterone, androstenedione (Adion), dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulfate (DHEAS), and sex hormone binding globulin (SHBG) in serum and peritoneal fluid. Three groups were studied, in which the peritoneal fluid was collected within 1, 3 and 5 days after ovulation. E1, E2 and progesterone levels in peritoneal fluid were highest shortly after ovulation and decreased with time. Testosterone and Adion in peritoneal fluid showed no changes, but peritoneal fluid levels were always higher than serum levels. No differences were found between the peritoneal fluid and serum levels of DHEA and DHEAS. SHBG in serum was always higher than in peritoneal fluid. The results are compared with reported steroid levels in follicular fluid from the literature and factors complicating the interpretation of steroid levels in peritoneal fluid are discussed.     

Effect of luteinized unruptured follicle cycles on clinical outcomes of frozen thawed embryo transfer in Chinese women

OBJECTIVE: To determine the effect of luteinized unruptured follicle (LUF) cycles on frozen thawed embryo transfer (FET). DESIGN: A retrospective analysis comparing the clinical outcomes after FET among 144 cases of luteinized unruptured follicle (LUF) cycles and 866 cases of ovulation cycles. SETTING: Reproductive medical center, Beijing China. PATIENTS: Chinese infertile women who underwent FET. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Clinical pregnancy rate (PR), implantation rate. RESULTS: The implantation rate, clinical pregnancy rate, on-going pregnancy rate and live birth rate in LUF group were 12.76% (49/384), 27.78% (40/144), 24.31% (35/144) and 19.44% (28/144), respectively, and in ovulation group, 14.74% (332/2251), 31.29% (271/866), 28.29% (245/866) and 22.23% (193/866), respectively (p > 0.05). CONCLUSIONS: LUF does not affect the clinical outcomes of FET. Patients of LUF should be included in FET treatment.     

Ultrasonographical study on luteinized unruptured follicle

The clinical features and the intrafollicular environment of luteinized unruptured follicle (LUF) were investigated in 60 infertile women by means of ultrasonography. LUF was diagnosed in daily ultrasound scans in 52 (13.5%) of 384 apparently ovulatory cycles. High incidences of LUF cycle were observed in the patients with polycystic ovary (37.5%), endometriosis (24.7%) and a history of pelvic surgery (26.2%), whereas LUF cycles were rare in the patients who conceived (2.7%). Mean follicular diameter measured by transvaginal ultrasonography showed no difference between LUF and ovulatory cycles during the follicular phase, whereas during the luteal phase, a continual growth pattern without follicular collapse was observed in LUF cycles. The ultrasonic aspiration of the follicle was carried out in 21 LUF cycles on the luteal phase. In 19 cases, hormonal profiles for the aspirated follicular fluid coincided with the preoperative diagnosis, and in 6 of 19 cases (31.6%), the presence of entrapped oocytes, direct evidence of LUF, was demonstrated. The morphological observation revealed that the oocytes recovered from the LUF follicle were in the stage of postmaturity, and the fertilization test in which they were used resulted in abnormal fertilization. These results indicate that LUF is an important cause of infertility and that periovarian abnormality might contribute to the failure of follicular rupture and postmaturity.