Age related semen parameters and ICSI pregnancy outcomes of 8046 men in Turkey over a 9-year period.

The effect of paternal age on fertility remains unclear. This retrospective study aims to examine the impact of male age on semen parameters and the reproductive outcomes of men admitted to an infertility center over a 9-year period. A total of 8046 patients were included in the study. Men were divided into four age groups. The groups were evaluated for semen parameters and reproductive outcome. The 21-30 year group presented lower sperm concentrations in comparison to those aged 31-40 and 41-50, yet shared a similar concentration to those over 50 years of age. Moreover, grades A and B decreased significantly in men aged over 50 years. The highest progressive motility and normozoospermia were observed in the age group 31-40 years while men over 50 years of age had the highest rates of asthenozoospermia and oligoasthenozoospermia. Furthermore, live birth results were reported in 5583 of the patients who underwent intracytoplasmic sperm injection (ICSI) and were found highest between 31-40 years of age. To our knowledge, this is the largest study in Turkey focusing on male age-related semen parameters and ICSI pregnancy outcomes. The study demonstrates that age is a significant factor for semen quality and live birth.

The Effects of Glutathione on Clinically Essential Fertility Parameters in a Bleomycin Etoposide Cisplatin Chemotherapy Model.

Chemotherapeutic agents used in the treatment of testicular cancer cause damage to healthy tissues, including the testis. We investigated the effects of glutathione on sperm DNA integrity and testicular histomorphology in bleomycin etoposide cisplatin (BEP) treated rats. Twelve-week-old male rats of reproductive age (n = 24) were randomly divided into three groups, the (i) control group, (ii) BEP group, and (iii) BEP+ glutathione group. Weight gain increase and testes indices of the control group were found to be higher than that of the BEP group and BEP+ glutathione group. While the BEP treatment increased sperm DNA fragmentation and morphological abnormalities when compared to the control group, GSH treatment resulted in a marked decrease for both parameters. Moreover, BEP treatment significantly decreased serum testosterone levels and sperm counts in comparison to the control group, yet this reduction was recovered in the BEP+ glutathione treated group. Similarly, seminiferous tubule epithelial thicknesses and Johnsen scores in testicles were higher in the control and BEP+ glutathione groups than in the BEP-treated group. In conclusion, exogenous glutathione might prevent the deterioration of male reproductive functions by alleviating the detrimental effects of BEP treatment on sperm quality and testicular histomorphology.

Early Compaction Might Be a Parameter to Determine Good Quality Embryos and Day of Embryo Transfer in Patients Undergoing Intracytoplasmic Sperm Injection.

INTRODUCTION: Compaction is the first event in embryo morphogenesis. Blastocyst transfer on day five or six has been widely performed in the last decade. We investigated the clinical value of early compaction on day three for evaluation of the transferred embryo quality and pregnancy. METHODS: Four hundred patients with female factor infertility and 776 fresh embryo transfers were included. Two groups were formed: Early compaction group had embryo transfer with at least one day-three embryo exhibiting early compaction. Transferred embryos without early compaction comprised the control group. Embryo transfer was performed on day three or five after the assessment of embryo compaction by a time-lapse technology system. Each patient underwent only a single cycle of embryo transfer. We analyzed fertilization, pregnancy, and live birth rates. RESULTS: We detected significantly higher numbers of the retrieved oocytes, metaphase II (MII) oocytes, and fertilized oocytes in the early compaction group. Moreover, the transfer of the early compacting embryos on day three resulted in higher pregnancy and live birth rates. CONCLUSION: Our data suggest that early compaction might be a factor to determine good quality embryos and embryo transfer day.

Rescue IVM of Denuded GV- and MI-Stage Oocytes of Premenopausal Rats with Oncostatin M, Insulin-like Growth Factor I, and Growth Hormone.

Immature oocytes are retrieved and matured through in vitro maturation (IVM). Maturation, fertilization rates, and embryo development via IVM are all lower than those found in vitro fertilization (IVF) cycles. We investigated the effects of oncostatin M (OSM), insulin-like growth factor-1 (IGF-I), and growth hormone (GH) in rescue IVM. A total of 111 germinal vesicle (GV) and 17 metaphase I (MI) oocytes were obtained after conventional IVF from 28 female Wistar albino rats. Denuded immature oocytes were cultured in maturation medium supplemented with OSM, IGF-1, or GH. The quantities of metaphase II (MII) oocytes matured from the GV stage were 17 of 30 (56.6%), 15 of 28 (53.5%), 10 of 30 (33.3%), and 7 of 23 (30.3%), in control, OSM, IGF-I, and GH groups, respectively. Maturation rates in control and OSM groups were higher than those in IGF-I and GH groups (p = 0.001). The quantities of MII oocytes matured from MI stage were 7 of 7 (100%), 4 of 4 (100%), 1 of 1 (100%), and 1 of 5 (20%) in control, OSM, IGF-I, and GH groups, respectively. Maturation rates from MI to MII stages in control, OSM, and IGF-I groups were higher than those in the GH group (p = 0.004). Acceptable maturation rates are observed with OSM in rat oocytes in rescue IVM.

Comparison of the rates for reaching the blastocyst stage between normal and abnormal pronucleus embryos monitored by a time-lapse system in IVF patients

Objective: To compare the rates of blastocyst stage development between embryos fertilized after one (MPN) or more than two pronucleus (PN) (3PN, 4PN-multiPN) with those after 2PN in the same patients. Material and Methods: The embryos of patients who had both abnormal PN (MPN, 3PN or 4PN) and normal fertilized (2PN) embryos after intracytoplasmic sperm injection (ICSI) fertilization, were followed with a time-lapse system following the ICSI procedure. The rates of reaching the blastocyst stage were compared between normal and abnormally fertilized embryos. Results: One thousand eight hundred and twenty oocytes were collected from 140 patients and 1280 (70.3%) of them were fertilized. MPN, 2PN and 3PN, 4PN (multiPN) ratios of the embryos in the pronuclear stage were 11.4%, 83.13% and 5.47%, respectively. The rates of reaching the blastocyst stage among these embryos were 17.1%, 60.8% and 42.8% for MPN, 2PN and multiPN, respectively. The proportion reaching blastocyst development was significantly higher following 2PN compared to those after MPN and multiPN (p<0.05). Embryos developing after multiPN had significantly higher rates of reaching the blastocyst stage compared to those after MPN (p<0.01). Conclusion: The majority of abnormally pronucleated embryos arrest without reaching the blastocyst stage. MultiPN embryos have a higher rate of blastocyst development than MPN embryos.

Sperm Selection and Embryo Development: A Comparison of the Density Gradient Centrifugation and Microfluidic Chip Sperm Preparation Methods in Patients with Astheno-Teratozoospermia.

In recent years, microfluidic chip-based sperm sorting has emerged as an alternative tool to centrifugation-based conventional techniques for in vitro fertilization. This prospective study aims to compare the effects of density gradient centrifugation and microfluidic chip sperm preparation methods on embryo development in patient populations with astheno-teratozoospermia. In the study, the semen samples of the patients were divided into two groups for preparation with either the microfluidic or density gradient methods. Selected spermatozoa were then used to fertilize mature sibling oocytes and the semen parameters and embryo development on days 3 and 5 were assessed. While the density gradient group was associated with a higher sperm concentration, motility (progressive and total) was significantly higher in the microfluidic chip group. No significant differences were observed in the fertilization rates or grade 1 (G1) and grade 2 (G2) proportions of the third-day embryos. Furthermore, while the proportions of the poor, fair and good blastocysts on day 5 did not differ significantly, excellent blastocysts (indicating high-quality embryos) were observed in a significantly higher proportion of the microfluidic chip group. When compared to the classical density gradient method, the microfluidic chip sperm preparation yielded sperm with higher motility and higher quality blastocysts at day 5; in patients with astheno-teratozoospermia.

Intracranial injection with KCl: an alternative method in selected cases of multifetal pregnancy reduction.

Multifetal pregnancy reduction (MFPR) offers a therapeutic option which reduces the maternal, prenatal, neonatal morbidity and mortality associated with multifetal pregnancies. In certain cases of MFPR, where difficulty is encountered in reaching the thorax due to the fetal position as well as the location of membranes and placenta, an alternative approach may be the insertion of the needle to the fetal cranium. We describe a new technique for MFPR performed by fetal intracranial injection of potassium chloride. To our knowledge, the current case series is the first report describing the technique of intracranial injection of potassium chloride during MFPR and selective termination. This approach enables a technically easier procedure than the intrathoracic approach. However, the use of this technique should be reserved for selected cases of MFPR only by experienced operators and centers.

IVF outcomes after hysteroscopic metroplasty in patients with T- shaped uterus.

BACKGROUND: T- shaped uterus may be associated with infertility and adverse pregnancy outcomes. Hysteroscopic metroplasty may improve the reproductivity for these cases. To our knowledge, there is no data in literature about the clinical consequences of in vitro fertilization (IVF) in patients undergoing hysteroscopic metroplasty for T-shaped uterus. The principal objective of the current study is to assess the impact of hysteroscopic metroplasty for T-shaped uterus on the reproductive outcomes of IVF. METHODS: IVF outcomes of 74 patients who underwent hysteroscopic metroplasty for T- shaped uterus and 148 patients without any uterine abnormalities and with diagnosis of unexplained infertility (control group) were retrospectively analyzed. RESULTS: Patients in metroplasty and control groups were comparable with respect to age, BMI, partner's age and duration of infertility. Number of patients with a history of pregnancy beyond 20 weeks of gestation was significantly lower in the metroplasty group (4.1% vs 18.2%; p < 0.05). Number of previous unsuccessful cycles and percentage of patients with ≥3 unsuccessful IVF cycles (35.1% vs 17.6%; p < 0.05) were significantly higher in the metroplasty group. There were no significant differences in the reproductive outcomes such as the pregnancy rate, clinical pregnancy or live birth rate between the metroplasty and control groups. There were non-significant trends for higher rates of miscarriage (18.8% vs 8%, p > 0.05) and biochemical pregnancy (20.0% vs 10.7%, p > 0.05) in the metroplasty group compared to the control group. CONCLUSIONS: Reproductive results of the IVF cycles after hysteroscopic correction of T-shaped uterus were comparable to those of the patients without any uterine abnormalities and with diagnosis of unexplained infertility. Hysteroscopic metroplasty may contribute to improved IVF outcomes in patients with T-shaped uterus.

Evaluation of Sperm DNA Fragmentation via Halosperm Technique and TUNEL Assay Before and After Cryopreservation.

AIM: Human sperm DNA fragmentation is one of the factors suggested for male infertility. The ratio of sperm DNA damage in semen may adversely affect both the fertilization rate and the embryo development of in vitro fertilization/ intracytoplasmic sperm injection cycles. Sperm cryopreservation both increases the success rates in assisted reproductive techniques (ARTs) and contributes to the preservation of fertility before testis surgery, chemotherapy, and radiotherapy. The aim of the current study is to determine sperm DNA fragmentation, following cryopreservation. METHODS: A cross-sectional, observational study was conducted at a university hospital infertility clinic. One hundred (n = 100) volunteer fertile men (ages between 21 and 39 years) with normozoospermic sperm parameters were involved in the current study. Sperm DNA damage was evaluated with the Halosperm technique and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay. Fresh samples were studied in liquid form. The remaining samples were kept frozen and then thawed after 1 month and reevaluated with the Halosperm technique and TUNEL assay. Results were then compared between the fresh and frozen samples. RESULTS: Sperm DNA fragmentation results with the Halosperm technique both before and after cryopreservation were 25% (5%-65%) and 40% (6%-89%), respectively, with a statistically significant increase (15%; P < .001). Sperm DNA fragmentation results by TUNEL assay before and after cryopreservation were 17% (3%-43%) and 36% (7%-94%), respectively, with a statistically significant increase (19%; P <.001). CONCLUSION: The current data demonstrate increased sperm DNA damage after cryopreservation. Further studies may contribute to development of less harmful techniques and cryoprotectants in order to improve the results of ART.

Estradiol increases apoptosis in human coronary artery endothelial cells by up-regulating Fas and Fas ligand expression.

CONTEXT: In animal models, estrogen inhibits atherogenesis by inhibiting many of the early steps of atherosclerotic plaque formation. However, the lack of cardioprotective effect by postmenopausal hormone replacement therapy and possible increase in cardiovascular events observed during the first year after the initiation of hormone replacement therapy may suggest that once the plaque is formed, estrogen may have additional effects that may counteract its beneficial outcomes. Indeed, the effect of estrogen on plaque stability has not been identified. OBJECTIVE: We hypothesized that 17beta-estradiol (E2) may cause increased apoptosis in human coronary artery endothelial cells (HCAECs). This effect would explain an adverse effect on plaque stability in vivo. INTERVENTION(S) AND MAIN OUTCOME MEASURE(S): The effect of E2 on apoptosis, cell proliferation, and expression of proapoptotic molecules Fas and Fas ligand (FasL) in cultured HCAECs was evaluated. RESULTS: HCAECs in culture treated with E2 showed an increase in DNA strand breaks and nuclear fragmentation indicative of apoptosis. E2 treatment also induced a significant concentration-dependent increase in Fas mRNA and protein expressions in HCAECs. Moreover, the expression of FasL mRNA and secretion of FasL protein by HCAECs were enhanced in response to E2 treatments. CONCLUSIONS: E2 increases the apoptosis in cultured HCAECs. Enhanced Fas and FasL expressions in response to E2 suggest that activation of the Fas/FasL pathway may be a mediator of the proapoptotic effects of E2 in these cells.

Serum and follicular fluid levels of soluble Fas and soluble Fas ligand in IVF cycles.

OBJECTIVE: To determine follicular fluid (FF) and serum levels of soluble Fas (sFas) and soluble Fas ligand (sFasL) in patients undergoing IVF cycles. STUDY DESIGN: A prospective comparative study among patients with endometriosis (n=12), infertility due to male factor (n=12) and poor responders (n=32) undergoing IVF cycles in Centrum IVF Clinic. Individual FF and serum samples were collected from patients during transvaginal ultrasonography-guided follicle aspiration. Patients were classified as poor responder patients undergoing IVF cycles with GnRHa, triptorelin and GnRH antagonist, cetrotide, patients with endometriosis and patients with infertility due to male factor. sFas, sFasL levels in both FF and serum samples and their correlations with clinical outcomes of IVF were measured in each study group. RESULTS: Serum and FF levels of sFas, sFasL were similar in the poor responder and male factor groups. There were no differences between the serum and FF levels of both sFas and sFasL among poor responder patients receiving either GnRH agonist or antagonist therapies. Serum levels of sFas were significantly lower in the endometriosis group compared to the male factor group. Serum and FF levels of sFas, sFasL were similar among patients with or without clinical pregnancy. CONCLUSION: sFas and sFasL are detected in both serum and follicular fluid samples from IVF cycles, their levels are similar between poor responder and male factor groups as well as between GnRH agonist and antagonist treatment groups. These soluble apoptotic factors may not be predictive for the outcomes of IVF. Decreased serum levels of sFas, suggests increased apoptosis in endometriosis.

Regulation of HOXA-10 expression by testosterone in vitro and in the endometrium of patients with polycystic ovary syndrome.

Polycystic ovary syndrome (PCOS) affects approximately 5% of reproductive-age women and is characterized by anovulation and increased androgen production. Despite the ability to correct ovulatory disorders, pregnancy rates remain paradoxically low, and spontaneous pregnancy loss rates are high. To determine whether uterine dysfunction contributed to the adverse reproductive outcomes in PCOS, we assessed the effect of the increased ovarian androgens on a well-characterized gene essential to endometrial receptivity. Up-regulation of HOXA10 in the endometrium is necessary for receptivity to embryo implantation. In vitro, HOXA10 expression was repressed by testosterone but not by dehydroepiandrosterone, dehydroepiandrosterone sulfate, or insulin. Testosterone also prevented the increased expression of HOXA10 previously reported with estradiol or progesterone. Dihydrotestosterone produced an effect similar to that of testosterone, whereas flutamide blocked the testosterone effect. Endometrial biopsies, obtained from women with PCOS, demonstrated decreased HOXA10 mRNA. Testosterone is a novel regulator of HOXA10. Diminished uterine HOXA10 expression may contribute to the diminished reproduction potential of women with PCOS.

Regulation of fas ligand expression by IL-8 in human endometrium.

Numerous cytokines and growth factors are synthesized in the endometrium. IL-8 is one of these cytokines regulating endometrial function. It is a neutrophil chemoattractant/ activating factor and a potent angiogenic agent. IL-8 is elevated in the peritoneal fluid of women with endometriosis. We have previously demonstrated a direct proliferative effect of IL-8 on endometrial stromal cells. We hypothesized that increased levels of IL-8 in the endometriotic environment could up- regulate Fas ligand (FasL) expression in endometrial cells and may be relevant for the development of a relative local immunotolerance in endometriosis by inducing apoptosis of cytotoxic T lymphocytes. To test our hypothesis, we studied the in vitro regulation of FasL expression and apoptosis by IL-8 in endometrial cells. Western blot analysis in endometrial stromal, glandular, and Ishikawa cells revealed that IL-8 up- regulated FasL protein expression in these cells. By semiquantitative RT-PCR analysis, IL-8 does not alter the expression of either Fas or FasL mRNA levels in these cells. Immunocytochemistry results from endometrial stromal cells treated with IL-8 demonstrated an up-regulation of FasL protein expression. IL-8 decreased apoptosis rate in endometrial stromal cells as evaluated by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling assay. We observed an increased apoptotic rate in Jurkat (T lymphocyte line) cells plated on endometrial stromal cells previously treated with IL-8. We speculate that increased FasL expression by IL-8 may induce apoptosis of T lymphocytes and thus produce a local immunotolerant environment for the development of ectopic implants.

Endometrial HOXA10 expression after controlled ovarian hyperstimulation with recombinant follicle-stimulating hormone.

OBJECTIVE: To determine the effects of controlled ovarian hyperstimulation and resultant high levels of E(2) on endometrial HOXA10 expression (a marker of endometrial receptivity). DESIGN: Prospective study. SETTING: University academic medical center. PATIENT(S): Twenty-five women undergoing controlled ovarian hyperstimulation with recombinant FSH and 30 fertile controls. INTERVENTION(S): Endometrium was obtained by Pipelle endometrial biopsy on cycle days 21-25. In addition, Ishikawa cells (a well-differentiated endometrial adenocarcinoma cell line) were treated with either E(2), recombinant FSH, GnRH agonist, or GnRH antagonist. RNA was extracted and analyzed by quantitative reverse transcriptase-polymerase chain reaction (RT-PCR). MAIN OUTCOME MEASURE(S): HOXA10 expression. RESULT(S): Endometrial HOXA10 expression in women undergoing controlled ovarian hyperstimulation (COH) with recombinant FSH was not different from that in fertile controls. Estradiol increased HOXA10 expression in Ishikawa cells in a dose-dependent manner from 10(-9) to 10(-7) M. Neither recombinant FSH, GnRH agonist, nor GnRH antagonist altered HOXA10 expression in these cells. CONCLUSION(S): Controlled ovarian hyperstimulation did not inhibit endometrial HOXA10 expression in vivo. In addition, in vitro endometrial cell HOXA10 expression was not altered by either recombinant FSH, GnRH agonist, or GnRH antagonist. COH is unlikely to adversely impact endometrial receptivity.

Estradiol down-regulates MCP-1 expression in human coronary artery endothelial cells.

OBJECTIVE: To determine whether estrogen down-regulates MCP-1 in vascular endothelial cells. DESIGN: A prospective comparative study. SETTING: Academic research environment. PATIENT(S): Human umbilical vein endothelial cells (n = 3) and human coronary artery endothelial cells (n = 3) obtained from females. INTERVENTION(S): Human umbilical vein endothelial cells (HUVEC) and human coronary artery endothelial cells (HCAEC) were grown to preconfluence. Then, they were treated with various concentrations of estradiol (10(-11) M to 10(-7) M) as well as raloxifene (10(-7) M) and tamoxifen (10(-7) M). MCP-1 in culture media was quantified using an enzyme-linked immunosorbent assay (ELISA). Cellular ribonucleic acid (RNA) was extracted and Northern blots were hybridized with an oligonucleotide probe complementary to a specific sequence of MCP-1 mRNA. MAIN OUTCOME MEASURE(S): MCP-1 protein and mRNA. RESULT(S): Estrogen treatment did not change MCP-1 expression in HUVEC. On the other hand, in HCAEC, estradiol induced a 30% decrease in mRNA expression and resulted in dose-dependent inhibition of MCP-1 production as detected by ELISA. Raloxifene and tamoxifen also resulted in inhibition of MCP-1 mRNA and protein expression. CONCLUSION(S): Our findings suggest that one of the mechanisms by which estrogen down-regulates atherosclerosis is by suppressing vascular MCP-1 expression, resulting in decreased macrophage recruitment.

Transient osteoporosis of pregnancy: case report.

Transient osteoporosis of pregnancy is a rarely observed skeletal pathology developing in the last months of pregnancy. Meticulous evaluation is important for the differential diagnosis of severe and progressive hip and/or groin pain in pregnant patients. MRI is a valuable and safe technique for demonstrating bone marrow edema and skeletal abnormalities during pregnancy. Avoidance of vaginal delivery and non-weight bearing measures are essential in order to prevent complications such as hip fractures related to transient osteoporosis of pregnancy. We present the diagnostic evaluation and treatment of an uncommon case of transient osteoporosis of pregnancy with resolution of symptoms and postpartum.

Laparoscopic management of heterotopic cesarean scar pregnancy with preservation of intrauterine gestation and delivery at term: case report.

OBJECTIVE: To present a case of laparoscopic removal of a heterotopic cesarean scar pregnancy under ultrasound guidance. DESIGN: Case report. SETTING: Private hospital. PATIENT(S): A 34-year-old woman with heterotopic cesarean scar pregnancy. INTERVENTION(S): Laparoscopic removal of heterotopic cesarean scar pregnancy. MAIN OUTCOME MEASURE(S): Delivery at term after laparoscopic management of heterotopic cesarean scar pregnancy. RESULT(S): An ongoing intrauterine pregnancy ended with a live birth after successful removal of the heterotopic gestational mass by a laparoscopic approach. CONCLUSION(S): Surgical removal of the ectopic mass by laparoscopy may be a radical approach in cases of heterotopic cesarean scar pregnancy. Laparoscopic excision of the cesarean scar pregnancy gives the opportunity to preserve the viable intrauterine gestation while maintaining a strong lower uterine segment. Ultrasound is an adjunctive tool that enables precise location of the ectopic mass during the operation.

Efficiency and pregnancy outcome of serial intrauterine device-guided hysteroscopic adhesiolysis of intrauterine synechiae.

OBJECTIVE: To highlight the efficiency of intrauterine device (IUD) guidance during hysteroscopic adhesiolysis for severe intrauterine adhesions. DESIGN: A prospective, randomized trial. SETTING: Private tertiary and referral infertility clinic. PATIENT(S): Seventy-one subfertile patients who underwent hysteroscopic treatment of intrauterine synechiae or adhesions. INTERVENTION(S): Thirty-six women in group 1 were initially examined by laparoscopy-hysteroscopy at first look, and an IUD was inserted during hysteroscopic adhesiolysis. The adhesions were further lysed by the guidance of IUD during the second-look office hysteroscopy, 1 week later. Patients were prescribed 2 months of estrogen as well as P therapy, and the IUD was removed by the end of this period. The uterine cavity was evaluated, and adhesions were further lysed by a third-look office hysteroscopy, 1 week after the removal of IUD. Thirty-five women in group 2 were similarly examined by first-look office hysteroscopy, and an IUD was inserted during hysteroscopic adhesiolysis. These patients did not undergo early intervention of office hysteroscopy, 1 week after the first procedure. They also used 2 months of estrogen and P therapy. The IUD was removed by the end of this period, and the uterine cavity was evaluated and adhesions were further lysed during a second-look office hysteroscopy. MAIN OUTCOME MEASURE(S): Pregnancy rate and live birth rate. RESULT(S): Spontaneous pregnancy rates after treatment were 17/36 (47.2%) and 11/35 (30%), and live birth rates were 10/36 (28%) and 7/35 (20%) in groups 1 and 2, respectively. These differences between the two groups were not statistically significant. CONCLUSION(S): The method described especially for early intervention may prevent complications during the treatment of severe intrauterine adhesions and may present a secure and effective alternative for constructive clinical outcomes.

Regulation of FAS ligand expression by chemokine ligand 2 in human endometrial cells.

Human endometrium is a dynamic tissue under the influence of numerous hormones, growth factors, and cytokines interacting to maintain a balance of cellular growth, differentiation, and apoptosis. We have previously demonstrated that several factors including interleukin-8, extracellular matrix, and steroid hormones modulate FASLG, one of the apoptotic molecules, in human endometrium. Chemokine ligand 2 (CCL2), a monocyte chemoattractant and activating factor, is a cytokine involved in endometrial function. CCL2 is elevated in the peritoneal fluid of women with endometriosis. We hypothesize that increased levels of CCL2 in the endometriotic environment may upregulate FASLG expression in human endometrial stromal cells and induce a local immunotolerance in endometriosis. To test our hypothesis, we studied the in vitro regulation of FASLG expression and apoptosis by CCL2 in endometrial stromal cells. Western blot analysis revealed that CCL2 upregulated FASLG protein expression in cultured endometrial stromal cells. Based on semiquantitative RT-PCR analysis, CCL2 did not alter either FAS or FASLG mRNA expression in endometrial stromal cells. Immunocytochemistry results from the same cells treated with CCL2 demonstrated upregulation of FASLG protein expression. CCL2 did not change rate of apoptosis in endometrial stromal cells as evaluated by TUNEL assay. However, an increased apoptotic rate was detected in Jurkat (T lymphocytes) cells cocultured with endometrial stromal cells previously treated with CCL2. We speculate that increased FASLG expression by CCL2 may induce apoptosis of T lymphocytes and thus produce an immunotolerant environment for the development of ectopic implants.

Impaired immunity against tetanus in type 2 diabetes.

BACKGROUND: The aim of this study was to compare serum levels of tetanus antibody in diabetic patients over 50 years of age with those of age- and sex-matched non-diabetic controls. MATERIAL/METHODS: The study population consisted of 115 type 2 diabetic patients and 115 age- and sex-matched non-diabetic patients. Serum levels of tetanus IgG were measured by a commercial ELISA kit, and levels over 0.1 IU/ml were considered protective. RESULTS: Mean serum levels of tetanus antibody in the diabetic and control groups were 0.164+/-0.140 IU/ml vs. 0.374+/-0.534 IU/ml, respectively (p<0.001). Mean serum levels of tetanus antibody in the diabetics vs. controls aged 50-64 years were 0.172+/-0.141 IU/ml vs. 0.568+/-0.653 IU/ml and in those p<0.001, p=1.000). Among patients aged 50-64 years, 38 (55.9%) cases in the diabetic and 45 (73.8%) in the control group demonstrated protective levels of tetanus antibodies (p=0.034). Of patients p=0.298). CONCLUSIONS: Serum levels of tetanus antibody decreased in diabetic patients older than 50 years of age, whereas this period of time is prolonged to 65 years in healthy individuals. All individuals over 65 years should be vaccinated against tetanus; however, vaccination over 50 years of age might be considered for diabetic patients.