Effect of physiotherapy on infertility treatment in polycystic ovary syndrome patients.

Introduction: Polycystic ovary syndrome (PCOS) is a complex hormonal condition associated with psychological, reproductive, and metabolic features. Low-grade inflammation is a recognised factor causing metabolic and reproductive disorders in PCOS, which is why anti-inflammation approaches in PCOS treatment, especially during the COVID pandemic, are considered. A promising therapeutic option is osteopathic manipulative treatment (OMT), which activates the cholinergic anti-inflammatory pathway and can inhibit proinflammatory cytokines, such as TNF, IL-1ß, and TNF-ß. In our paper we analysed the influence of OMT in women with PCOS. Material and methods: Seventy-three patients, aged 29-46 years, with a history of reproductive failure, who underwent a physiotherapeutic treatment were evaluated. Six months after the end of a physiotherapy session, a follow-up questionnaire was performed. Results: The results show that most women (83.6%) were satisfied with the therapeutic process, and that the sessions met the patient's expectations. Reducing the level of anxiety related to infertility treatment and pregnancy was declared by 60 (82.2%) women, an improvement in well-being was declared by 72 (97.3%) (p = 0.04), and increasing awareness of the body after physiotherapy sessions was declared by 70 (95.9%) women. Conclusions: The physiotherapeutic sessions improved infertility treatment, enhanced quality of life, and had a positive effect on overall health in PCOS women.

Preliminary Results for Personalized Therapy in Pregnant Women with Polycystic Ovary Syndrome During the COVID-19 Pandemic.

Increased androgen level, hyperinsulinemia, diabetes, impaired fibrinolysis, obesity, hypertension, chronic inflammation, abnormal immune response to infections and hyperhomocysteinemia are the most common abnormalities related to polycystic ovary syndrome (PCOS) women and are the factors predisposing to the severe course of COVID-19. The SARS-Cov-2 infection during pregnancy is associated with an increased risk of complications (spontaneous abortion), similar to those in PCOS. The treatment of PCOS pregnant women with a history of fertility failures raises many doubts, especially during the COVID pandemic. However, due to the increasing incidence of infections among reproductive people and the potentially more serious course in pregnant women, numerous questions about the safety and effectiveness of the treatment are still very current. In our study we presented a series of cases of recurrent miscarriages or recurrent implantation failure PCOS pregnant women with confirmed COVID-19. The diagnosis of infertility confirmed the presence of plasminogen activator inhibitor type 1 and/or 5,10-methylenetetrahydrofolate reductase polymorphisms in each of them. Moreover, some of the women presented immune dysfunction associated with infertility. We have described the personalized treatments of each pregnant patient included: metformin, enoxaparin and tacrolimus. The treatment applied had the expected effect, supporting the implantation processes. Furthermore, despite the ambiguous data according to immunological therapy of infertile women during the COVID pandemic, we observed a mild or asymptomatic COVID-19 course and we noticed no pregnancy complications.

Sildenafil Citrate Downregulates PDE5A mRNA Expression in Women with Recurrent Pregnancy Loss without Altering Angiogenic Factors-A Preliminary Study.

In our previous study, we showed that sildenafil citrate (SC), a selective PDE5A blocker, modulated NK cell activity in patients with recurrent pregnancy loss, which correlated with positive pregnancy outcomes. It was found that NK cells had a pivotal role in decidualization, angiogenesis, spiral artery remodeling, and the regulation of trophoblast invasion. Thus, in the current study, we determined the effects of SC on angiogenic factor expression and production, as well as idNK cell activity in the presence of nitric synthase blocker L-NMMA. Methods: NK cells (CD56+) were isolated from the peripheral blood of 15 patients and 15 fertile women on MACS columns and cultured in transformation media containing IL-15, TGF-ß, and AZA-a methylation agent-for 7 days in hypoxia (94% N2, 1% O2, 5% CO2). Cultures were set up in four variants: (1) with SC, (2) without SC, (3) with NO, a synthase blocker, and (4) with SC and NO synthase blocker. NK cell activity was determined after 7 days of culturing as CD107a expression after an additional 4h of stimulation with K562 erythroleukemia cells. The expression of the PDE5A, VEGF-A, PIGF, IL-8, and RENBP genes was determined with quantitative real-time PCR (qRT-PCR) using TaqMan probes and ELISA was used to measure the concentrations of VEGF-A, PLGF, IL-8, Ang-I, Ang-II, IFN-γ proteins in culture supernatants after SC supplementation. Results: SC downregulated PDE5A expression and had no effect on other studied angiogenic factors. VEGF-A expression was increased in RPL patients compared with fertile women. Similarly, VEGF production was enhanced in RPL patients' supernatants and SC increased the concentration of PIGF in culture supernatants. SC did not affect the expression or concentration of other studied factors, nor idNK cell activity, regardless of NO synthase blockade.

Decreased Production of TNF-α and IL-6 Inflammatory Cytokines in Non-Pregnant Idiopathic RPL Women Immunomodulatory Effect of Sildenafil Citrate on the Cellular Response of Idiopathic RPL Women.

Sildenafil citrate (SC), a PDE5 inhibitor, a drug for erectile dysfunction (ED) and pulmonary hypertension (PAH), was found to exert a positive effect on pregnancy outcomes when administered intravaginally before conception. In our previous studies, sildenafil increased endometrial thickness and significantly decreased peripheral blood NK cell activity after the intravaginal administration in women with recurrent pregnancy loss (RPL). No data are available to confirm the effect of sildenafil on maternal T cell populations involved in shaping fetal-maternal tolerance and NK cell activity. Thus, the present study aimed to establish if SC influences NKT cells or the axis of Th17/Treg cells and Th1/Th2 cytokine production. MATERIALS AND METHODS: Twenty-one healthy fertile women and twenty-two nonpregnant women with idiopathic RPL were studied. The ELISA method was used to evaluate the production of cytokines, including IL-2, IL-12p40, IL-4, IL-10, IL-6, IL-17, IL-21, TGF-ß, TNF-α, and IFN-γ in PBMC culture supernatants before and after supplementation with the physiological concentration of SC. The percentages of NKT (CD56+CD3+CD44+CD161+), Treg (CD4+CD25+FOXP3+) and Th17 (CD4+CD25+IL-17A+) cells were determined with flow cytometry method. RESULTS: Unexpectedly, we found that the PBMCs of patients with RPL produced a significantly lower level of inflammatory cytokines (TNF-α and IL-6) and a higher level of anti-inflammatory cytokines (TGF-ß and IL-10). SC significantly decreased IL-6, IL-12 and increased TGF-ß cytokine concentration in fertile women. In the case of RPL patients' PBMCs, SC improved the production of TNF-α and IL-10. CONCLUSIONS: Lower concentration of proinflammatory cytokines in idiopathic RPL women compared to fertile women might suggest the exhaustion of the immune system. The emphasized production of IL-10 by SC partially explains the previously observed downregulation of NK cell activity in RPL patients. The immunomodulatory effect of the drug might be utilized in anti-inflammatory therapies and help achieve positive pregnancy outcomes in women with reproductive failure due to a Th1/Th2 imbalance.

Differences in the Expression of KIR, ILT Inhibitory Receptors, and VEGF Production in the Induced Decidual NK Cell Cultures of Fertile and RPL Women.

RESULTS: KIR2DL1 and ILT-2 expression on idNK cells was higher in healthy women than in RPL patients. Sildenafil enhanced NKG2A expression in RPL patients. VEGF concentration was higher in fertile woman idNK cell cultures. idNK cells were more sensitive for necrosis in RPL than in fertile women. SC did not influence VEGF production or idNK cell apoptosis. CONCLUSIONS: A combination of hypoxia, IL-15, and AZA promotes the conversion of pbNK into idNK cells CD56+CD16--expressing KIR receptors and produces VEGF. Alterations in KIR2DL1 and ILT-2 expression as well as impaired VEGF production were associated with RPL. SC affects NKG2A expression on RPL idNK cells. SC had no effect on VEGF release or idNK cell apoptosis.

Anti-Müllerian hormone level is associated with vitamin D receptor polymorphisms in women with polycystic ovary syndrome.

OBJECTIVE: Our study aimed to investigate the relationship between polymorphisms (Apa1, Bsm1, Fok1, and Cdx2) in the VDR gene as well as AMH and AMHR2 genes and their influence on AMH and 25(OH)D levels in PCOS women. STUDY DESIGN: Seventy-five patients with PCOS and 23 control women were included. Serum AMH and 25(OH)D levels in patients and controls were measured by enzyme-linked immunosorbent assay (ELISA). Polymorphisms in VDR gene Fok1 C/T (rs2228587), Bsm1 A/G (rs1544410), Apa1 A/C (rs7975232), and Cdx2 A/G (rs11568820) polymorphisms as well as AMH G/T (rs10407022) and AMHR2 A/G (rs2002555) were analyzed using real-time PCR. RESULTS: Analysis of the VDR Cdx2 polymorphism showed a significantly higher frequency of the homozygous GG (mutant) genotype in the PCOS group as compared with the control group (p < 0.05). The analysis revealed a statistically significant correlation between the presence of FokI and ApaI polymorphisms and AMH levels in PCOS women (p < 0.05). The presence of mutant genotypes (CT, TT) in the Fok1 and (CA, CC) in the Apa1 polymorphisms were associated with higher AMH level in PCOS women (p < 0.05). No statistically significant correlations between AMH and AMHR2 polymorphisms and AMH level were found. Moreover, there was no correlation between AMH and 25(OH)D levels in the PCOS or in the control group. CONCLUSION: It seems that the elevated AMH level is associated with VDR Fokl and Apal polymorphisms, but not with 25(OH)D levels in PCOS women. Further research is needed to determine the role of VDR polymorphism in AMH level in PCOS.

Content of fatty acids in a diet and the homocysteine levels in women with fertility disorders.

OBJECTIVES: Assessment of the effects of consumption of fatty acids on the homocysteine levels in women with fertility disorders. METHODS AND RESULTS: The study included 286 women at the age between 23 and 46 years (the mean 33.13±4.21 years) with a fertility disorders. We measured: levels of homocysteine [µmol/L] (n=171), body weight and height (n=286). Body mass index (BMI) was calculated. A diet was assessed by 3-day dietary food records method including one day of the weekend. The mean homocysteine (Hcy) levels were 10.02±2.98 µmol/L. Body weight excess was observed in 29.3% of subjects. The percentage of fatty acids in the total energy [%E] it was demonstrated that the mean rate of MUFAs and PUFAs was statistically significantly lower in diets of women with the homocysteine levels >15 µmol/L compared to the group with the Hcy <10 µmol/L. Based on the results of a correlation between the homocysteine levels and consumption of n-6 fatty acids and n-3 fatty acids it is possible to conclude that there are negative correlations observed indicating that higher consumption of these acids is associated with lower levels of homocysteine. CONCLUSION: Increased consumption of PUFAs, including α-linoleic acid, in a diet seems to be an important factor preventing from hyperhomocysteinemia.

3D laparoscopy as a tertiary cytoreductive (TCR) surgery in infiltrating ureter recurrent epithelial ovarian cancer - case report and a mini-review of the literature.

The paper describes a case of a 61-year-old woman with recurrent epithelial ovarian cancer infiltrating the ureter treated with 3D laparoscopy as a tertiary cytoreductive surgery (TCR). In addition, a mini-review of the literature concerning TCR is presented.

Decreased NK cell activity after partial vulvectomy in pregnant patient with vulvar intraepithelial neoplasia 3 (VIN3).

INTRODUCTION AND OBJECTIVE: The study presents the problem of immune disturbances in pregnant women with vulvar carcinoma in situ (VIN3). MATERIAL AND METHODS: NK cell and T reg activity in the study patient were analysed using flow cytometry. RESULTS: Decreased NK cell activity and but increased T reg activity were observed after vulvectomy, with subsequent successful pregnancy outcome. CONCLUSIONS: Although vulvar cancer may influence immune cell activity, this issue merits further study.

Methylenotetrahydrololate reductase A1298C and C677T polymorphisms and adverse pregnancy outcome in women with PCOS.

OBJECTIVES: The aim of this study was to compare SNP C677T and A1298C in the MTHFR gene and pregnancy outcome in PCOS women. STUDY DESIGN: We investigated 76 PCOS and 56 non-PCOS women. Among PCOS patients 63 were women with a history of recurrent pregnancy loss (RPL) and 13 women were infertile. In non-PCOS group 40 women were RPL and 16 were infertile. We investigated the relationship between SNP in the MTHFR gene and pregnancy loss, homocysteine and AMH concentration in the study groups. RESULTS: DNA analysis of the PCOS and non-PCOS groups for MTHFR C677T and A1298C polymorphism showed no significant association between the groups. We demonstrated an increased miscarriage rate in non-PCOS women with A1298C polymorphism in the MTHFR gene (p=0.042). We found that homocysteine concentration was higher in women with SNP MTHFR A1298C (p=0.046). Moreover, we did not observe any association between the level of homocysteine and the pregnancy outcome in the whole study group. CONSLUSION: It seems that the presence of the MTHFR mutation is not associated with PCOS in the Polish population. However, our results may suggest a correlation between the MTHFR A1298C mutation and RPL in the non-PCOS group.

Successful pregnancy after Intralipid addition to sildenafil and enoxaparin in woman with history of recurrent pregnancy loss (RPL).

OBJECTIVES: Does addition of Intralipid to sildenafil and enoxaparin immunotherapy improve pregnancy outcome? MATERIALS AND METHOD: Report of a striking case of a patient with history of 4 recurrent pregnancy losses (RPL) and IVF failures. RESULTS: Adding of Intralipid resulted in giving birth to a healthy male baby in the 3th IVF cycle. CONCLUSION: Combination therapy that includes Intralipid may generate successful IVF outcome, although this problem merits further study, especially regarding safety issues.

Effects of sildenafil citrate and etanercept treatment on TNF-α levels in peripheral blood of women with recurrent miscarriage.

OBJECTIVES: The aim of the study was to determine serum concentrations of a proinflammatory cytokine, tumor necrosis factor-alpha (TNF-α), in patients with recurrent abortions undergoing treatment with sildenafil or etanercept. MATERIAL AND METHODS: Serum TNF-α concentrations were determined for 24 patients with recurrent miscarriages (aged 32.7 ± 4.64 years) deemed eligible for sildenafil therapy and 7 patients treated with etanercept (aged 37.65 ± 5.45 years). Measurements were performed before and after therapy. The control group included 10 healthy women (aged 33.3 ± 5.49 years), who gave birth at least once without pregnancy-related complications. The levels of serum TNF-α were measured by Elisa. RESULTS: Patients treated with etanercept had significantly elevated levels of TNF-α before therapy as compared to the control group (41.4 ± 28.4 vs. 16.6 ± 7.2 pg/ml). Moreover we found a tendency for the concentration of TNF-α to increase in sera of patients treated with sildenafil after therapy completion (19 ± 29 vs. 15.4 ± 26.7 pg/ml). Treatment with etanercept resulted in a significant reduction of serum TNF-α levels (41.4 ± 28.4 vs. 25.4 ± 3.2 pg/ml). CONCLUSIONS: Therapy of recurrent abortions with anti-TNF-α drugs appears to be encouraging. Administration of blockers of phosphodiesterase type 5 or TNF-α blockers before conception seems to be a promising future therapy of immune-dependent recurrent miscarriages, limiting the teratogenic influence of the drugs on the fetus.

Retrospective comparison between the Prolift and Elevate anterior vaginal mesh procedures: 18-month clinical outcome.

INTRODUCTION AND HYPOTHESIS: There are few direct comparisons between the first-generation trocar-guided and the second-generation single-incision mesh systems in the treatment of anterior pelvic organ prolapse (POP). Hence, the purpose of this retrospective review was to compare 18-month operative success in female patients who had undergone POP surgery with the anterior Prolift (n = 52) or the anterior Elevate mesh (n = 62). METHODS: Subjective (bulge symptoms) and objective measures (absence of anterior or apical descent beyond the hymen, POP-Q anterior stage 0 or I, no retreatment for POP) were used as the measures of surgical efficacy. Postoperative pelvic floor pain, dyspareunia, de novo overactive bladder (OAB), de novo stress urinary incontinence (SUI), and mesh exposure were addressed as complications of POP surgery. RESULTS: The two groups did not differ with regard to the subjective and objective measures of the operative efficacy. There were no between-group differences in the proportion of women reporting postoperative pelvic floor pain, dyspareunia, de novo SUI, and de novo OAB symptoms (all p values >0.05). The proportion of patients with postoperative vaginal exposure was significantly higher in the Prolift group (7.7 %) than in the Elevate group (0.0 %; p = 0.02). CONCLUSIONS: In conclusion, our results suggest that the use of the Elevate system in patients with anterior compartment prolapse results in fewer mesh erosions, but similar efficacy, compared with the Prolift mesh.

Anti-Müllerian hormone concentration as a biomarker of pregnancy success or failure.

OBJECTIVE: This study was conducted to determine serum anti-Müllerian hormone (AMH) concentration influence on pregnancy outcome. STUDY DESIGN: In this study we investigated sixty one infertile women (aged 27 to 44 years) who were diagnosed and treated between 2011 and 2013. We determine ovarian reserve measured by AMH concentration. Patients were divided in three groups according to their serum AMH concentration (<1 ng/ml; 1-2.5 ng/ml; >2.5 ng/ml respectively). We investigated the relationship between clinical pregnancy rate and AMH concentration. In addition, anti-thyroid antibodies (anti-TG and/or anti-TPO) positivity and insulin concentration were correlated with AMH level and pregnancy outcome in the study groups. RESULTS: We found no statistical differences between AMH concentration regarding number of pregnancies (42.3%; 41.1 %; 38.9% respectively in study groups; p>0.05). The miscarriage rate was highest in women with AMH>2.5 ng/mL (27.3%, 0%, 86% respectively in study groups; p>0.05). We found that anti-thyroid positivity is more frequent in women with lower AMH concentration (23.1%; 11.7%; 5.5% respectively; p>0.05) and patients with lower serum AMH had higher serum insulin concentration (p<0.05). CONCLUSIONS: It seems that AMH concentration might not reflect oocyte quality and the chance of pregnancy, but increased AMH concentration may be associated with negative pregnancy outcome. Moreover, it cannot be excluded that presence of anti-thyroid antibodies and increased insulin serum concentration may be connected to diminished ovarian reserve measured by AMH concentration.

Survivin expression as a prognostic factor in patients with epithelial ovarian cancer or primary peritoneal cancer treated with neoadjuvant chemotherapy.

BACKGROUND: The aim of this study was to evaluate association of expression of survivin and p53 with the effects of neoadjuvant chemotherapy (NAC) in patients with advanced ovarian cancer (AOC). METHODS: We retrospectively evaluated 60 consecutive patients with AOC (International Federation of Gynecology and Obstetrics stage IIIC-IV) treated with NAC. The expression of p53 and survivin was assessed immunohistochemically. The median of expression total score survivin equals 2 was adopted to dichotomize the group. The positive and negative expression of p53 was used to dichotomize the group. RESULTS: The expression of survivin in tumor tissue taken before and after NAC was a significant difference in the percentage of stained nuclei (P = 0.0002), the intensity of staining (P = 0.0003), and total score (P = 0.0001). There was a significant difference in p53 expression in tumor tissue before and after NAC in the percentage of stained nuclei (P = 0.0424). Survivin expression, in contrast to p53 expression, was a prognostic factor in patients with AOC treated with NAC (P = 0.0484). The expression of survivin and p53 was not a predictive factor. Independent adverse predictor factors were as follows: lack of optimal interval debulking surgery and the lack of an objective response (the respective hazard ratio was 3.93 [95% confidence interval, 2.07-7.46; P < 0.0001] and 2.36 [95% confidence interval,1.25-4.47; P = 0.0080]). The suboptimal range of interval debulking surgery, resistance to platinum, and the lack of paclitaxel in the NAC were adverse prognostic factors (the respective hazard ratio was 2.61 [95% confidence interval, 1.17-5.83], 2.72 [95% confidence interval, 1.07-6.89], and 2.56 [95% confidence interval, 1.06-6.18]; P < 0.05]). CONCLUSIONS: High expression of survivin could be a prognostic factor in patients treated with NAC for AOC.

[Ovarian aging and infertility]. / Procesy starzenia sie komórki jajowej a nieplodnosc.

The biological state of the ovum remains the key element in normal reproduction. Age-related decrease in the number of oocytes, as well as disturbed neuroendocrine function of the ovary and lesions in the uterus, contribute to reduced fertility. Decreasing number of ovarian follicles is accompanied by reduction of their quality including mainly abnormalities of the nucleus (dispersed chromatin, decondensation of chromosomes and abnormalities connected with the spindle apparatus). This results in failed reproduction due to abnormal gametogenesis, fertilization process, early development of the embryo and abnormal implantation. This work describes age-related biochemical mechanisms conditioning molecular changes occurring due to abnormal microenvironment of the ovary; their accumulation leads to aging and to a more rapid apoptosis of the oocyte. There are many theories explaining the causes of oocyte destruction, including abnormal vascularization, oxidative stress, imbalance of free radicals, influence of toxic compounds and genetic changes. Decreased blood perfusion in the microenvironment of a maturating ovum leads to hypoxia and thus to a chain of reactions of oxidative stress. Oxidative imbalance leads to abnormalities of cellular biomolecules. Moreover it is suggested that glycation processes in a cell, leading to the formation of compounds called AGEs (Advanced Glycation End Products), are also responsible for aging of the cells. They contribute directly to protein damage, induce a chain of reactions of oxidative stress, and increase the inflammatory reactions. Recently the role of mitochondria and telomeres in the aging process and loss of reproductive functions has been especially underscored. Moreover this work stresses the prognostic value of clinically used markers evaluating the ovarian reserve. The role of Anti-Müllerian hormone (AMH), follicle-stimulating hormone (FSH), estradiol, inhibin B and antral follicle count (AFC) was presented in this paper.

Etanercept immunotherapy in women with a history of recurrent reproductive failure.

OBJECTIVE: The aim of the study was to evaluate the effect of etanercept immunotherapy on peripheral natural killer (NK) cell activity in women with a history of recurrent miscarriage (RM) or failed in vitro fertilization (IVF). MATERIALS AND METHODS: Thirty nonpregnant women with reproductive failure and increased peripheral NK-cell number and/or activity before conception were studied. Women with reproductive failure received 4 doses (25 mg) of etanercept twice weekly before conception. Peripheral NK-cell activity before and after etanercept therapy in RM women was measured using flow cytometry In addition, the peripheral blood NK-cell surface antigens- CD16- and CD56 and peripheral blood regulatory T cell (T reg) antigens- CD4- and CD25 were studied using flow cytometry before treatment and 2 weeks after the last etanercept dose. RESULTS: NK-cell activity was significantly decreased after etanercept therapy in the study women (P < .05). This effect was significantly higher in women with subsequent pregnancy success (P < .05), but not in those with pregnancy failure (P > .05). There were no significant differences in T reg level before and after etanercept therapy (P > 0.05). CONCLUSION: Etanercept therapy might be effective treatment for women with increased NK-cell activity. Regulation of immune system activity may underlie the possible effect of such therapy.

First successful pregnancy after addition of enoxaparin to sildenafil and etanercept immunotherapy in woman with fifteen failed IVF cycles - case report.

PROBLEM: Does addition of enoxaparin to sildenafil and etanercept immunotherapy improve IVF outcome? METHODS: Report of a striking case with 15 IVF failures. RESULT: When enoxaparin was added, the 16th IVF cycle generated a healthy male baby. CONCLUSIONS: Combination therapy that includes a heparin may allow successful IVF outcome and this issue merits further study.

Reliability and clinical validity of a Polish version of the CONTILIFE: a quality of life questionnaire for urinary incontinence.

INTRODUCTION AND HYPOTHESIS: The primary aim of the study was to translate and validate a Polish version of the CONTILIFE, a quality of life questionnaire. The clinical validity of the CONTILIFE was assessed against the number of urinary leaks, Valsalva leak point pressure (VLPP), and maximum urethral closure pressure (MUCP). METHODS: One hundred women with stress urinary incontinence completed a Polish version of the CONTILIFE twice with a 4-day interval. RESULTS: Internal consistency (Cronbach's alpha= 0.94) and test-retest reliability of the instrument (ICC = 0.96) were very good. A significant relationship was found between all the CONTILIFE dimension scores, the CONTILIFE global score, and the number of urinary leaks per week. A less significant relationship was found between VLPP, MUCP, and CONTILIFE dimension and global scores. CONCLUSIONS: A Polish version of the CONTILIFE can be a reliable measure of quality of life in stress urinary incontinence patients.

Ectopic and eutopic stromal endometriotic cells have a damaged ceramide signaling pathway to apoptosis.

OBJECTIVE: To investigate whether sphingosine analogues, which activate the ceramide signaling pathway to apoptosis, can cause the death of ectopic (EEC) and eutopic stromal endometriotic cells (EEU), as well as healthy eutopic stromal endometrial cells (HEU). DESIGN: The EEC, EEU, and HEU isolated from fertile and infertile women with endometriosis were cultured for 48 hours in RPMI medium with 10% fetal calf serum (FCS) and with 2.5-10 microM sphingosine analogues. SETTING: A clinic for the treatment of endometriosis and basic research laboratories. PATIENT(S): Nineteen women with follicular cyst and 16 women with endometriosis. MAIN OUTCOME MEASURE(S): The percentage of proliferating cells was determined by 93-(4,5-dimethylthiazol-2-yl)2,5-diphenyl tetrazolium bromide (MTT) assay. Apoptosis and cell cycle were detected by fluorescence-activated cell sorter (FACS) Calibur flow cytometer. RESULT(S): The viability of EEC after exposure to 10 microM sphingosine analogues was 59.5% +/- 9.7% for D-sphingosine and 77.65 +/- 9.7% for DL-erythro-sphingosine, the viability of EEU was 69.2% +/- 14.2% and 42.0% +/- 15.5%, whereas the viability of comparative HEU was 9.0% +/- 4.8% and 18.8% +/- 8.3%, respectively. The differences were significant using the Mann-Whitney test. The apoptotic level of the cells treated with 10 microM sphingosine analogues for comparative HEU was 42.8% +/- 7.5% for D-sphingosine and 42.5% +/- 10.5% for DL-erythro-sphingosine, whereas for EEC this was 16.7% +/- 5.5% for D-sphingosine and 14.1% +/- 4.4% for DL-erythro-sphingosine and for EEU this was 14.3% +/- 4.7% and 22.9% +/- 8.9%, respectively. CONCLUSION(S): Ectopic and eutopic stromal endometrial cells from women with endometriosis have a damaged ceramide-downstream pathway to apoptosis.