Fertility and Sterility

Recurrent implantation failure: current update and clinical approach to an ongoing challenge

Neri Laufer, Alex Simon — 2012-05-01

A better understanding of the mechanisms responsible for implantation may improve our ability to treat RIF and improve IVF results in general. This issue's Views and Reviews aims to summarize the current knowledge on mechanisms involved in recurrent implantation failure and presents a clinical approach and potential treatments to overcome the problem.

Recurrent implantation failure: gamete and embryo factors

Mausumi Das, Hananel E.G. Holzer — 2012-03-16

Chromosomal abnormalities, sperm DNA damage, zona hardening, inadequate culture conditions, and suboptimal embryo development all play a significant role in the etiology of recurrent implantation failure. Evidence suggests that preimplantation genetic screening does not increase implantation or live birth rates. Comparative genomic hybridization array and analysis of single nucleotide polymorphisms could enable a more comprehensive screening of chromosomes. Assisted hatching may help to overcome zona hardening in selected cases. Optimal culture conditions and blastocyst transfer could contribute toward improving implantation and pregnancy rates. Novel embryo assessment and selection procedures, such as time-lapse imaging and metabolomics, may help in better evaluation of embryo quality and viability and help in selecting embryos with the highest implantation potential. The safety and efficacy of emerging treatment modalities should be evaluated in prospective randomized clinical trials before being applied in routine clinical practice.

Defective endometrial receptivity

Ariel Revel — 2012-05-01

The endometrium is one of the most fascinating tissues in the human body. Its sole purpose is to enable implantation of an embryo during a relatively short window of opportunity in the menstrual cycle. It is becoming clear that overcoming the current bottleneck in improvements to assisted reproductive techniques will require a closer look at the interface between uterus and embryo. Indeed, embryo implantation requires a cross talk with a receptive endometrium. Using sonography, hysteroscopy and endometrial biopsy we can learn about anatomical and functional markers of endometrial receptivity. This article reviews the factors which might cause defective endometrial receptivity. These include uterine polyps, septa, leiomyomata and adhesions. The effect of thin endometrium, endometriosis and hydrosalpinx is also described. Finally contemporary investigation of molecular markers of endometrial receptivity is described. Improving embryo implantation by a closer look inside the uterus is the key to increasing pregnancy rates in IVF.

Recurrent IVF failure: other factors

Alan S. Penzias — 2012-03-29

IVF failure is a problem for a couple in the singular but can be a tragedy in the plural. Recurrent IVF failure has multiple known causes but many which are not routinely considered as part of the posttreatment analysis. The reason is there are several causes associated with lifestyle and other causes related to pre-existing conditions that have only a tenuous or no apparent connection to fertility. This article examines the impact of obesity, cigarette smoke, uterine anatomy, body mass index, thyroid dysfunction, immune factors, the hereditary and acquired thrombophilias, and embryo transfer technique on recurrent IVF failure.

Repeated implantation failure: clinical approach

Alex Simon, Neri Laufer — 2012-04-01

Successful embryo implantation depends on a well-functioning endometrium as well as a normal healthy embryo. This process might be hampered if either of these variables is defective. Repeated implantation failure (RIF) is diagnosed when good-quality embryos repeatedly fail to implant after transfer in several IVF treatment cycles. The causes of RIF originate with either the mother or the embryo. The authors discuss factors that are associated with RIF and address various treatment options.

A persistent misperception: assisted reproductive technology can reverse the “aged biological clock”

Nichole Wyndham, Paula Gabriela Marin Figueira, Pasquale Patrizio — 2012-03-05

Delaying motherhood should be a free choice made in full knowledge of all the consequences, but modern women have alarming misconceptions about their own reproductive systems and the effectiveness of assisted reproductive technologies. Doctors and health professionals must begin to discuss fertility preservation with their patients and make sure that young women truly understand all their options. Preventing age-related infertility is the responsibility not only of doctors and medical practitioners but also of society at large. Social, economic, and personal pressures are causing women to decide to conceive later in life, yet those who choose to delay motherhood are stigmatized as being selfish and unconcerned about starting a family. This stigma must be banished, and age-related infertility should be faced as a medical problem.

A good meta-analysis is hard to find

Richard S. Legro, Allen R. Kunselman — 2012-05-01

“A good man is hard to find,” Red Sammy said. “Everything is getting terrible. I remember that day you could go off and leave your screen door unlatched. Not no more.”From A Good Man is Hard to Find by Flannery O’Connor

The incidence and effect of bacteriospermia and elevated seminal leukocytes on semen parameters

2012-02-17

Objective: To determine the incidence of bacteriospermia and elevated seminal leukocytes (ESL) in a subfertile male population and correlate these results with semen parameters.Design: Retrospective cohort study.Setting: Canadian tertiary-level male infertility clinic and university-affiliated andrology and microbiology laboratories.Patient(s): Four thousand nine hundred thirty-five nonazoospermic subfertile men.Intervention(s): Analysis and concurrent culture of 7,852 semen samples.Main Outcome Measure(s): Incidence of bacteriospermia and ESL and comparison of semen parameters between these groups.Result(s): The rate of bacteriospermia was 15% (22 species), and the rate of ESL was 19%, with no statistical correlation between these groups. Bacteriospermic patients (without ESL) had a statistically significant deterioration in DNA fragmentation index (DFI) only, compared with patients without bacteriospermia and ESL (24.1 vs. 21.8%). ESL alone was associated with a statistically significant deterioration in sperm concentration (20.6 vs. 55.3 × 106/mL), motility (21.8 vs. 26.9%), normal morphology (12.3 vs. 17.4%), and DFI (26.5 vs. 21.8%), with no additional deterioration identified with bacteriospermia.Conclusion(s): Bacteriospermia and ESL were prevalent, but not statistically associated, in subfertile men. Bacteriospermia alone was associated with an increase in DFI only, but the presence of ESL was the dominant factor associated with deterioration in semen parameters.

Electroejaculation combined with intracytoplasmic sperm injection in patients with psychogenic anejaculation yields comparable results to patients with spinal cord injuries

2012-02-27

Objective: To evaluate sperm quality and fertility potential of men with psychogenic anejaculation treated by electroejaculation (EEJ) and intracytoplasmic sperm injection (ICSI). Treatment results were compared to spinal cord injured (SCI) patients treated similarly.Design: Retrospective clinical study.Setting: Academic tertiary referral fertility center.Patient(s): Couples with isolated psychogenic anejaculation or SCI.Intervention(s): Electroejaculation and ICSI.Main Outcome Measure(s): Semen analysis, fertilization rate, implantation rate, pregnancy rate, delivery rate and safety of the procedure.Result(s): Fifteen patients diagnosed with psychogenic anejaculation underwent 40 EEJ/ICSI cycles. The semen retrieved was characterized by low motility (mean 10.7% ± 12.3%), normal volume (2.2 ± 1.9 mL) and normal count (25.1 ± 29.9 × 106/mL), according to World Health Organization criteria. Results of EEJ/ICSI were compared with 22 SCI patients treated by 66 EEJ/ICSI cycles during the same period. Mean female age and the number of oocytes retrieved per cycle were similar between the groups. Similar semen parameters after EEJ were found between psychogenic and SCI patients. Fertilization rate was significantly lower in the psychogenic patients compared to SCI (47.0% and 57.0%, respectively). No significant differences were found regarding pregnancy rates (20% and 22.7%, respectively), implantation rate (10.2% and 11.6%, respectively) or delivery rates (15% and 18.2%, respectively).Conclusion(s): Sperm retrieved by EEJ is characterized by asthenospermia and normal count. In spite of the lower fertilization rate in psychogenic patients, combination of EEJ and ICSI gives adequate results to couples with psychogenic anejaculation similar to the results obtained for SCI patients. Current results give these couples a reasonable chance of pregnancy achievement.

A platform for evaluating sperm RNA biomarkers: dysplasia of the fibrous sheath—testing the concept

Aletheia Lima-Souza, Ester Anton, Shihong Mao, Won Jin Ho, Stephen A. Krawetz — 2012-03-02

Objective: To investigate the potential use of correlative microarray-based transcript pairs as candidate markers for male fertility using dysplasia of the fibrous sheath (DFS) as an affected model. It is widely recognized that microarray technology may be limited by cost and that the quality of the transcript remains relatively unknown. To address these issues, we analyzed the stable transcript pairs by qPCR with a systematic primer design process.Design: Experimental study.Setting: University.Patient(s): Men with proven fertility and men with a diagnosis of DFS.Intervention(s): None.Main Outcome Measure(s): Primer sequences for six genes of interest were designed using Oligo7 and Primer3Plus. Primer specificity was initially assessed in silico by searching the ENSEMBL, University of California Santa Cruz, and National Center for Biotechnology Information databases for nontarget complementary sequences throughout the genome. The ability of transcript pairs to classify samples from males of proven fertility away from DFS was assessed.Result(s): In conjunction with identifying four new stable transcript pairs, comparison of the DFS qPCR C(t) correlation coefficients revealed the disruption of four stable fertile sample transcript pairs. This suite of transcript pairs resolves DFS.Conclusion(s): The results show that with effectively designed primers, qPCR may provide an affordable molecular assay to assess male fertility status.

DNA integrity is maintained after freeze-drying of human spermatozoa

2012-03-05

Objective: To evaluate the effects on human spermatozoa of freeze-drying, also known as lyophilization, and of cryopreservation in liquid nitrogen.Design: Prospective experimental study.Setting: Reproductive medicine unit and a private IVF center.Patient(s): Thirty healthy male donors.Intervention(s): Sperm samples from 30 donors divided as two aliquots, one to be lyophilized and the other to be cryopreserved in liquid nitrogen.Main Outcome Measure(s): Assessment of count, motility, morphology, viability, DNA integrity, chromosomal status, and birefringence properties of lyophilized and cryopreserved human spermatozoa compared with the same parameters in the fresh sample.Result(s): Although sperm viability and motility were totally compromised after freeze-drying, the sperm chromatin structure was not altered in comparison with fresh samples, which demonstrated that the procedure did not affect DNA integrity. The sperm-head inner protoplasmic structures were also preserved, which was estimated by assessing the corresponding birefringence characteristics. After cryopreservation with liquid nitrogen, the motility, viability, and DNA integrity of spermatozoa were statistically significantly reduced compared with the fresh samples; the proportion of sperm cells with abnormal head birefringence increased meaningfully.Conclusion(s): The process of freeze-drying deeply damages cell membranes; however, unlike with liquid nitrogen preservation, it does not affect DNA integrity.

Emergency testicular sperm extraction after scrotal trauma in a patient with a history of contralateral orchiopexy for cryptorchidism: case report and review of the literature

2012-03-12

Objective: To report a case of successful testicular sperm extraction performed during an emergency scrotal exploration for trauma in a patient with a history of previous contralateral cryptorchidism.Design: Case report.Setting: Clinica Urologica I, Università degli Studi di Milano, Fondazione Istituto di Ricerca e Cura a Carattere Scientifico Ca' Granda, Ospedale Maggiore Policlinico.Patient(s): A 28-year-old man with a history of right orchiopexy for cryptorchidism presenting with blunt trauma of the pelvis.Intervention(s): Seminiferous tubules extraction.Main Outcome Measure(s): Successful sperm retrieval.Result(s): Scrotal ultrasonography (US) showed a suspected rupture of the left tunica albuginea, and color Doppler examination evidenced testicular perfusion only in the region near the mediastinum testis. Twelve hours after scrotal exploration, the extracted testicular tissue was successfully processed, and seven vials of sperm were frozen.Conclusion(s): After testicular trauma in selected patients, such as individuals with a history of monorchism or cryptorchidism, emergency testicular sperm extraction (TESE) can be a useful option to preserve fertility.

An investigation into the relationship between the metabolic profile of follicular fluid, oocyte developmental potential, and implantation outcome

2012-02-23

Objective: To determine whether metabolomic analysis of follicular fluid could prove a useful noninvasive technique for the selection of viable oocytes and embryos.Design: Metabolomic analysis based on proton nuclear magnetic resonance (1H NMR) performed on follicular fluid collected from in vitro fertilization (IVF) patients.Setting: A university research center and a private fertility clinic.Patient(s): Fifty-eight women undergoing IVF treatment.Intervention(s): Follicular fluid collected at the time of oocyte retrieval.Main Outcome Measure(s): Metabolomic profile, assessment of oocyte developmental potential and embryo viability.Result(s): The metabolomic profile of follicular fluid from follicles where the oocyte resulted in a fertilized egg that failed to cleave (n = 9) was distinctly different from that where oocytes developed into early cleavage-stage embryos. Discriminating metabolites included glucose, lactate, choline/phosphocholine, and lipoproteins. Comparison of follicular fluid from women who subsequently had a positive β human chorionic gonadotropin (n = 10) to those who were unsuccessful in achieving a pregnancy (n = 12) revealed metabolic differences that were correlated to cycle outcome.Conclusion(s): Differences in the metabolite composition of follicular fluid correlate with the developmental competence of the human oocyte. Therefore, metabolomic profiling of follicular fluid may prove to be an important technique in gamete/embryo selection.

Effects of laser polar-body biopsy on embryo quality

2012-02-24

Objective: To evaluate the effect of laser polar-body biopsy (PBB) for preimplantation genetic diagnosis on embryo quality.Study design: Retrospective case-control analysis. The quality of 145 embryos after PBB was compared to 276 embryos of the same group of women without biopsy.Setting: University-based tertiary-care medical center.Patient(s): Women with inherited genetics disease.Intervention(s): Laser PBB of IVF embryos for genetic diagnosis.Main Outcome Measure(s): The study and control embryos were compared for fertilization rate, pronuclear grading, and cleavage-stage parameters on days 1, 2, and 3 after oocyte retrieval.Result(s): The study embryos demonstrated higher rates of cleavage arrest (3.6% vs. 0.7%), higher rate of significant fragmentation on day 2 (9.5% vs. 3.0%), and lower rate of good cleavage embryos on day 2 (69.1% vs. 78.4%) compared with control embryos. On day 3, the study embryos had lower cleavage rates (six or more blastomeres; 56.5% vs. 74.5%), higher fragmentation (11.7% vs. 3.9%), higher rate of embryos presenting inferior cleavage pattern (57.2% vs. 38.5%), and lower mean blastomere number (5.8 ± 2.1 vs. 6.6 ± 1.9) compared with control embryos.Conclusion(s): Polar-body biopsy may have a negative effect on embryo quality.

What is the optimal follicular size before triggering ovulation in intrauterine insemination cycles with clomiphene citrate or letrozole? An analysis of 988 cycles

Anna Palatnik, Estil Strawn, Aniko Szabo, Paul Robb — 2012-03-28

Objective: To determine the optimal size of the leading follicle before human chorionic gonadotropin (hCG) administration in cycles with clomiphene citrate (CC) and letrozole, and to examine any differences in the optimal leading follicle size between cycles with CC and letrozole.Design: A retrospective study.Setting: University hospital-based reproductive center.Patient(s): 1,075 women undergoing intrauterine insemination cycles with CC or letrozole.Intervention(s): Leading follicle diameters and endometrial thickness were recorded 24 hours before hCG administration, together with other cycles parameters, and were compared between pregnant and nonpregnant patients.Main Outcome Measure(s): Leading follicle diameter and intrauterine insemination outcome.Result(s): Eight percent of patients (n = 87) were excluded because their leading follicle was less than 18 mm by days 11 to 13. Pregnancy was recorded as clinical pregnancy with fetal heart activity seen at 6- to 7-week transvaginal ultrasound. For both CC and letrozole, higher pregnancy rates were achieved when the leading follicles were in the 23 to 28 mm range. The optimal size of the leading follicle was not statistically significantly different between cycles using CC or letrozole. However, for each endometrial thickness, the optimal follicular size of the leading follicle was different. Each additional millimeter of endometrial thickness increased the optimal follicular size by 0.5 mm. Thicker endometrial lining led to a higher probability of pregnancy.Conclusion(s): The optimal size of the leading follicle in ovulation induction with CC and letrozole is similar for both drugs and is closely related to the endometrial thickness.

How members of the Society for Reproductive Endocrinology and Infertility and Society of Reproductive Surgeons evaluate, define, and manage hydrosalpinges

2012-03-12

Objective: To describe the management of hydrosalpinges among Society for Reproduction Endocrinology and Infertility (SREI)/Society of Reproductive Surgeons (SRS) members.Design: Cross-sectional survey of SREI/SRS members.Setting: Academic and private practice–based reproductive medicine physicians.Participant(s): A total of 442 SREI and/or SRS members.Intervention(s): Internet-based survey.Main Outcome Measure(s): To understand how respondents evaluate, define, and manage hydrosalpinges.Result(s): Of 1,070 SREI and SRS members surveyed, 442 responded to all items, for a 41% response rate. Respondents represented both academic and private practice settings, and differences existed in the evaluation and management of hydrosalpinges. More than one-half (57%) perform their own hysterosalpingograms (HSGs), and 54.5% involve radiologists in their interpretation of tubal disease. Most respondents thought that a clinically significant hydrosalpinx on HSG is one that is distally occluded (80.4%) or visible on ultrasound (60%). Approximately one in four respondents remove a unilateral hydrosalpinx before controlled ovarian hyperstimulation (COH)/intrauterine insemination (IUI) and clomiphene citrate (CC)/IUI (29.3% and 22.8%, respectively), and physicians in private practice were more likely to intervene (COH: risk ratio [RR] 1.81, 95% confidence interval [CI] 1.31–2.51; CC: RR 1.98, 95% CI 1.33–2.95). Although laparoscopic salpingectomy was the preferred method of surgical management, nearly one-half responded that hysteroscopic tubal occlusion should have a role as a primary method of intervention.Conclusion(s): SREI/SRS members define a “clinically significant hydrosalpinx” consistently, and actual practice among members reflects American Society for Reproductive Medicine/SRS recommendations, with variation attributed to individual patient needs. Additionally, one in four members intervene before other infertility treatments when there is a unilateral hydrosalpinx present.

Oocyte slow freezing using a 0.2–0.3 M sucrose concentration protocol: is it really the time to trash the cryopreservation machine?

Veronica Bianchi, Michela Lappi, Maria Antonietta Bonu, Andrea Borini — 2012-02-23

Objective: To update results on outcomes with frozen/thawed oocytes using a differential sucrose concentration during dehydration (0.2 M) and rehydration (0.3 M), combined with a one-step propanediol exposure.Design: Retrospective cohort study.Setting: Private IVF centers.Patient(s): Infertile couples undergoing IVF treatment.Intervention(s): Oocyte thawing cycles between May 2004 and December 2010.Main Outcome Measure(s): Survival, fertilization, and cleavage rates were reported to evaluate biological outcomes. Clinical pregnancy and implantation rates were analyzed as markers of efficiency.Result(s): Three hundred forty-two patients and 443 cycles were monitored; the survival was 71.8%, fertilization 77.9%, and of the embryos obtained 83.8% were classified as grade 1 and 2. Three hundred ninety-four transfers were performed, resulting in 90 pregnancies. The pregnancy rate per transfer was 22.8% and per patient was 26.3%, with 122 gestational sacs. The implantation rate per embryo was 13.5%. Patients were divided into three groups according to their age: ≤34 years (group A), 35–38 years (group B), and ≥39 years (group C). Biological outcomes were comparable in all three groups, whereas the pregnancy rate per transfer was higher in the first group (27.7% vs. 21.4% and 17.6%). The implantation rates per injected egg were 11.8%, 8.0%, and 7.5% for the three groups, respectively.Conclusion(s): The biological and clinical data obtained on 443 cycles are consistent with our previous results showing that slow freezing of oocytes can be a valid tool in IVF practice when performed with a suitable protocol.

The use of recombinant luteinizing hormone in patients undergoing assisted reproductive techniques with advanced reproductive age: a systematic review and meta-analysis

2012-02-27

Objective: To evaluate the effect of recombinant LH in assisted reproduction technology (ART) cycles in patients of advanced reproductive age.Design: A systematic review and meta-analysis.Setting: Published randomized controlled clinical trials comparing recombinant LH plus recombinant FSH versus recombinant FSH only in patients of advanced reproductive age.Patient(s): Patients 35 years and older undergoing assisted reproduction.Intervention(s): Recombinant LH plus recombinant FSH controlled ovarian hyperstimulation (COH) versus recombinant FSH stimulation only in assisted reproduction cycles.Main Outcome Measure(s): Implantation and clinical pregnancy.Result(s): Seven trials were identified that met inclusion criteria and comprised 902 assisted reproduction technology cycles. No differences in serum E2 on the day of hCG administration were reported in any trials. Two trials reported lower oocyte yield and one trial reported lower metaphase II oocyte yield in the recombinant LH-supplemented group. One trial reported higher fertilization rates in the recombinant LH-supplemented group. In a fixed effect model, implantation was higher in the recombinant LH-supplemented group (odds ratio 1.36, 95% confidence interval 1.05–1.78). Similarly, clinical pregnancy was increased in the recombinant LH-supplemented group (odds ratio 1.37, 95% confidence interval 1.03–1.83).Conclusion(s): The addition of recombinant LH to ART cycles may improve implantation and clinical pregnancy in patients of advanced reproductive age.

Why does the fallopian tube fail in ectopic pregnancy? The role of activins, inducible nitric oxide synthase, and MUC1 in ectopic implantation

2012-03-16

Objective: To investigate the role of activin-βA subunit, activin type II receptors, inducible nitric oxide synthase (iNOS), and MUC1 in the pathogenesis of ectopic pregnancy (EP) and their involvement in the determination of the implantation site.Design: Observational study.Setting: Academic unit of reproductive and developmental medicine.Patient(s): Four women at the luteal phase, three pseudopregnant women at the time of hysterectomy for benign disease, and 10 archived cases of EP. We collected 14 Fallopian tubes were collected from four women at the luteal phase and three pseudopregnant women at the time of hysterectomy for benign disease; specimens from implantation site, trophoblast and remote sites from the implantation site were collected from 10 archived cases of EP.Intervention(s): Immunohistochemistry and quantitative reverse-transcriptase polymerase chain reaction (RT-PCR).Main Outcome Measure(s): Comparison of the expression of candidate molecules between the different groups.Result(s): The expression of activin-βA subunit, activin type II receptors, and iNOS was statistically significantly increased and expression of MUC1 statistically significantly decreased in tubes bearing an EP. There was no statistically significant difference in the expression of the candidate molecules between the implantation and remote sites. Candidate molecules were also expressed in the trophoblast.Conclusion(s): The pathological expression of candidate molecules by tubes bearing an EP is not involved in the determination of implantation site. Additionally, candidate molecules may play a role in the regulation of trophoblast cells in vivo during early pregnancy.

Genetic association study of polymorphisms FOXP3 and FCRL3 in women with endometriosis

2012-02-17

Objective: To consider a possible cumulative effect of two genetic polymorphisms (FOXP3 C-2383T/rs3761549 and FCRL3 C-169T/rs7528684) that were previously shown to be associated with endometriosis.Design: Genetic association study.Setting: Human reproduction outpatient clinic of Faculdade de Medicina do ABC.Patient(s): One hundred eighty-eight infertile women with endometriosis and 169 controls.Intervention(s): Detection of polymorphisms FOXP3 (C-2383T/rs3761549) and FCRL3 (C-169T/rs7528684) by TaqMan real-time polymerase chain reaction. The results were analyzed statistically.Main Outcome Measure(s): Genotype distribution, allele frequency, and combination analysis of the FOXP3 and FCRL3 polymorphisms.Result(s): Single-marker analysis revealed a significant association of FOXP3 C-2383T and FCRL3 C-169T, independently, with endometriosis-related infertility, regardless of the stage of the disease. Considering the combined genotypes of FCRL3 and FOXP3 polymorphisms, a positive association was found between genotypes FCRL3TT/FOXP3CT, FCRL3CT/FOXP3CT, and FCRL3CC/FOXP3CT and the risk of endometriosis development. Moreover, a progression of the disease risk was observed according to the presence of one or two copies of risk allele FCRL3 C and only one copy of risk allele FOXP3 T (odds ratio [OR] = 2.14, OR = 3.25, and OR = 6.0, respectively, for genotypes FCRL3TT/FOXP3CT, FCRL3CT/FOXP3CT, and FCRL3CC/FOXP3CT).Conclusion(s): Our findings support a possible gene-gene interaction leading to a cumulative effect on endometriosis development.

Increased expression of macrophage colony–stimulating factor and its receptor in patients with endometriosis

2012-02-27

Objective: To investigate the expression and regulation of colony-stimulating factor 1 (CSF-1) and its receptor, C-FMS, in endometriosis.Design: In vivo and vitro study.Setting: University-based academic medical center.Patient(s): Reproductive-age women undergoing surgery for benign conditions.Intervention(s): Peritoneal and endometrial tissue samples were obtained.Main Outcome Measure(s): CSF-1 and C-FMS expression.Result(s): Significantly higher CSF-1 levels were found in peritoneal fluid of patients with endometriosis compared with control subjects. Ectopic endometriotic tissue had 3.5-fold and 1.7-fold increases in CSF-1 and C-FMS expression, respectively, compared with eutopic tissue. Coculture of endometrial cells from either established cell lines or patient samples with peritoneal mesothelial cells (PMCs) led to increased expression of CSF-1 and C-FMS. A higher but nonsignificant increase in levels of C-FMS and CSF-1 was found in cocultures of endometrial epithelial cells from patients with endometriosis compared with those without endometriosis.Conclusion(s): Increased CSF-1 levels may contribute to endometriosis lesion formation and progression. Elevation in CSF-1 after coculture of endometrial cells with PMCs suggests that endometrial tissue may be a source of peritoneal CSF-1. Increased C-FMS expression in endometrial cells from women with endometriosis cocultured with PMCs suggests that endometrial tissue involved in lesion formation is highly responsive to CSF-1 signaling.

A prospective cohort study of physical activity and time to pregnancy

2012-03-16

Objective: To investigate the association between leisure-time physical activity (PA) and fecundability.Design: Prospective cohort study.Setting: Internet-based observational study of Danish women who were planning a pregnancy (2007–2009).Patient(s): A total of 3,628 women aged 18–40 years at baseline.Intervention(s): None.Main Outcome Measure(s): Time to pregnancy (TTP). Fecundability ratios (FRs) and 95% confidence intervals (CIs) were derived from discrete-time Cox models, with adjustment for potential confounders, such as body mass index (BMI).Result(s): We observed an inverse monotonic association between vigorous PA and fecundability (≥5 h/wk vs. none: FR 0.68, 95% CI 0.54–0.85) and a weak positive association between moderate PA and fecundability (≥5 vs. <1 h/wk: FR 1.18, 95% CI 0.98–1.43) after mutual adjustment for both PA types. Inverse associations between high vigorous PA and fecundability were observed within subgroups of age, parity status, and cycle regularity, but not among overweight or obese women (BMI ≥25 kg/m2).Conclusion(s): There was evidence for a dose-response relationship between increasing vigorous PA and delayed TTP in all subgroups of women with the exception of overweight and obese women. Moderate PA was associated with a small increase in fecundability regardless of BMI. These findings indicate that PA of any type might improve fertility among overweight and obese women, a subgroup at higher risk of infertility. Lean women who substitute vigorous PA with moderate PA may also improve their fertility.

Glutathione transferase polymorphisms and risk of endometriosis associated with polychlorinated biphenyls exposure in Italian women: a gene–environment interaction

2012-03-15

Objective: To investigate the occurrence of a gene–environment interaction between glutathione transferase (GST) gene polymorphisms (GSTM1, GSTT1, GSTP1, and GSTA1) and serum polychlorinated biphenyls (PCBs) levels. This is suggested as possible risk factors for endometriosis, a multifactorial gynecological disease.Design: Case-control study conducted from 2002 to 2005.Setting: Policlinico Umberto I, “Sapienza” University of Rome and Italian National Institute for Health, Rome.Patient(s): Italian women (N = 343), with laparoscopic diagnosis and histologic confirmation of the presence (cases, N = 181) or the absence (controls, N = 162) of endometriosis.Intervention(s): Genomic DNA extraction, multiplex polymerase chain reaction (PCR), and restriction fragment length polymorphism analysis. Determination of serum concentrations of selected PCBs by ion-trap mass spectrometry (subgroup, 63 cases and 63 controls).Main Outcome Measure(s): Endometriosis diagnosis by laparoscopy, GST genotypes, serum PCB levels.Result(s): The genotype distributions of GSTM1, GSTA1, and GSTP1 did not show any statistically significant difference between cases and controls. The GSTT1 null genotype was negatively associated with the disease. The GSTP1 wild-type genotype in the presence of medium-high blood levels of PCB153, total PCBs, or of high levels of PCB180 significantly increased the risk of endometriosis, suggesting a multiplicative interaction.Conclusion(s): The GSTs polymorphisms per se do not increase the risk of developing endometriosis. However, a gene–environment interaction was observed for GSTP1Ile/Ile and GSTM1 null genotypes, modulating the effect of PCB153, PCB180, and of total PCBs on disease risk.

Orthotopic grafting of cryopreserved prepubertal testicular tissue: in search of a simple yet effective cryopreservation protocol

2012-02-27

Objective: To investigate whether solid-surface vitrification (SSV) is an effective cryopreservation strategy regarding the integrity and function of prepubertal mouse testicular tissue.Design: Prospective experimental study.Setting: Academic research unit.Animal(s): Mice.Intervention(s): Testicular tissue from 5- to 10-day-old GFP+ mice was cryopreserved with the use of a conventional uncontrolled slow freezing (USF) technique and SSV before intratesticular grafting in busulfan-treated GFP− mice.Main Outcome Measure(s): Ultrastructural cryoinjury to spermatogonial stem cells (SSCs) and somatic cells was assessed by electron microscopy. Tubular structure was evaluated by histology, and graft survival and spermatogenic recovery by immunohistochemistry.Result(s): The tubular morphology and the proportion of ultrastructural cryodamage were similar between vitrified and slow-frozen testicular fragments. Allografting of tissue after both USF and SSV resulted in a recovery of spermatogenesis similar to fresh samples.Conclusion(s): SSV resulted in success rates similar to USF in maintaining testicular cell ultrastructure, tubular morphology, and tissue function. These data provide further evidence that vitrification, being an inexpensive and simple technique, can be considered as an alternative for cryopreservation of prepubertal testicular tissue.

Microarray expression profiling in adhesion and normal peritoneal tissues

2012-02-23

Objective: To identify molecular markers associated with adhesion and normal peritoneal tissue using microarray expression profiling.Design: Comparative study.Setting: University hospital.Patient(s): Five premenopausal women.Intervention(s): Adhesion and normal peritoneal tissue samples were obtained from premenopausal women. Ribonucleic acid was extracted using standard protocols and processed for hybridization to Affymetrix Whole Transcript Human Gene Expression Chips. Microarray data were obtained from five different patients, each with adhesion tissue and normal peritoneal samples. Real-time polymerase chain reaction was performed for confirmation using standard protocols.Main Outcome Measure(s): Gene expression in postoperative adhesion and normal peritoneal tissues.Result(s): A total of 1,263 genes were differentially expressed between adhesion and normal tissues. One hundred seventy-three genes were found to be up-regulated and 56 genes were down-regulated in the adhesion tissues compared with normal peritoneal tissues. The genes were sorted into functional categories according to Gene Ontology annotations. Twenty-six up-regulated genes and 11 down-regulated genes were identified with functions potentially relevant to the pathophysiology of postoperative adhesions. We evaluated and confirmed expression of 12 of these specific genes via polymerase chain reaction.Conclusion(s): The pathogenesis, natural history, and optimal treatment of postoperative adhesive disease remains unanswered. Microarray analysis of adhesions identified specific genes with increased and decreased expression when compared with normal peritoneum. Knowledge of these genes and ontologic pathways with altered expression provide targets for new therapies to treat patients who have or are at risk for postoperative adhesions.

A novel luteinizing hormone/chorionic gonadotropin receptor mutation associated with amenorrhea, low oocyte yield, and recurrent pregnancy loss

Yaakov Bentov, Shlomit Kenigsberg, Robert F. Casper — 2012-02-27

Objective: To study the cause for poor oocyte yield, amenorrhea, and recurrent pregnancy loss in a patient undergoing IVF.Design: Case report.Setting: University-affiliated private IVF clinic.Patient(s): A 33-year-old woman with amenorrhea, recurrent ovarian cyst formation, poor oocyte yield, and repeated chemical pregnancies after IVF treatments.Intervention(s): The hCG stimulation test and luteinizing hormone/chorionic gonadotropin receptor (LHCGR) gene sequencing.Main Outcome Measure(s): The presence of LHCGR gene mutations.Result(s): The patient had a markedly abrogated androgen response to 10,000 IU of hCG. A novel heterozygous inactivating mutation in exon 1 of the LHCGR gene was detected. This mutation was superimposed on a common LHCGR polymorphism.Conclusion(s): This novel mutation may provide a potential genetic mechanism for the poor oocyte recovery in some IVF cases. It is the first example of a heterozygous inactivating mutation in the LHCGR gene.

Assessment of FSHR variants and antimüllerian hormone in infertility patients with a reduced ovarian response to gonadotropin stimulation

2012-03-09

Objective: To study women with a poor response to ovarian hormone stimulation, known as low responders. Genetic defects in the FSH receptor gene (FSHR) were analyzed as well as antimüllerian hormone (AMH) for ovarian reserve.Design: Retrospective cohort study.Setting: University hospital.Patient(s): Two hundred fifty-nine patients total: 74 low responders; 88 patients receiving assisted reproduction therapy (ART) with a normal ovarian response; and 97 women with a normal fertility status.Intervention(s): DNA from patients was analyzed using real-time polymerase chain reaction. Serum concentrations of AMH were assessed using ELISA.Main Outcome Measure(s): The FSHR variants Asn680Ser (rs6166), Ala189Val (rs121909658), Ile160Thr (rs121909659), Thr449Ile (rs28928870) and the serum AMH concentrations were assessed.Result(s): With the exception of the frequent Asn680Ser polymorphism, no homozygotic SNPs of FSHR were found. In the group of ART patients, Thr160/Ile160 variants were more frequent in comparison with women with normal fertility. The Ser680/Ser680 was more frequent in ART patients than in women with normal reproductive function. The rate of live births was markedly reduced, particularly in the low responder group. No difference was noted in the distribution of the Ala189Val and Thr449Ile variant. Low serum AMH values were observed in 75% of the low responder group.Conclusion(s): FSHR gene variations such as Asn680Ser, Ala189Val, Thr449Ile, and Ile160Thr did not seem to be a decisive factor of poor response to fertility treatment, whereas the low ovarian reserve determined by AMH is considered more crucial.

Viability and function of the cryopreserved whole rat ovary: comparison between slow-freezing and vitrification

Milan Milenkovic, César Diaz-Garcia, Ann Wallin, Mats Brännström — 2012-02-17

Objective: To investigate four different protocols for cryopreservation of the whole rat ovary with intact vasculature to evaluate whether differences exist in post-thawing viability of the ovary after either vitrification or slow freezing.Design: Experimental study.Setting: Obstetrics and gynecology department.Animal(s): Immature Sprague-Dawley female rats.Intervention(s): Ovaries were isolated with the vascular tree intact up to the bifurcation of the abdominal aorta and were subsequently cannulated. The ovaries were flushed with increasing concentrations of the cryoprotectant dimethyl sulfoxide (DMSO) to either 1.5 or 7 M. The ovaries underwent cryopreservation by vitrification or passive slow freezing.Main Outcome Measure(s): After thawing, the ovaries were subjected to neutral red viability staining to assess the density of viable small follicles and for long-term (48 hours) incubation evaluation of steroid secretion, histology, and apoptosis assay.Result(s): The follicular viability was decreased in both vitrification groups and in the slow-freezing group with the high concentration of DMSO, as compared with fresh controls. Estradiol levels in the incubation medium followed the same pattern. Light microscopy revealed well-preserved morphology in all groups after 48 hours’ incubation. Apoptosis was increased in both vitrified and cryopreserved ovaries.Conclusion(s): We have developed a new method that can be used in basic studies to improve cryopreservation protocols. Our initial findings suggest that a moderate concentration of the cryoprotectant DMSO is superior to a high DMSO concentration for both vitrification and slow freezing.

The impairment of reproduction in db/db mice is not mediated by intraovarian defective leptin signaling

2012-02-16

Objective: To demonstrate whether leptin modulates reproduction by a direct effect within the ovary.Design: Animal model.Setting: National Key Laboratory of Infertility.Animal(s): Adult female db/db mice.Intervention(s): Adult littermate wild-type (WT) and diabetic (db) leptin receptor (LR) mutant female mice were matched for the allograft of the ovary to construct new genotypic models, respectively. WT mouse received only one ovary from a WT or a db/db mouse (WT Ov-WT, WT Ov-db), and db/db mouse received one ovary from a WT or a db/db mouse (db Ov-WT, db Ov-db). WT and db/db mice received one ovary from a WT mouse and another ovary from a db/db mouse (WT Ov-WT/db, db Ov-WT/db) or received two ovaries all from a WT mouse (db Ov-WT/WT).Main Outcome Measure(s): Hormones, lipids, and reverse transcription polymerase chain reaction.Result(s): Both WT Ov-WT and WT Ov-db mice presented normal cycles, comparable serum E2 and FSH levels, and ovarian expressions of the Star, Cyp17, and Cyp19 mRNA, even with different ovary genotypes. In WT Ov-WT/db with hMG stimulation, db ovaries with LR mutation expressed higher Star, Cyp17, Cyp19, Jak2, Stat3, and Pias3 mRNA than in the basal state, whereas WT ovaries with intact LR expressed higher Star, Cyp17, and Cyp19 but divergently lower Jak2, Stat3, and Pias3 levels.Conclusion(s): We confirmed that impairment of reproduction in intact db/db mice is not mediated by intraovary intact/defective leptin signaling even in face of a divergent modulation by gonadotropins.

Bioluminescence imaging as a tool to evaluate germ cells in vitro and transplantation in vivo as fertility preservation of prepubertal male mice

2012-03-15

Objective: To determine the feasibility of bioluminescence imaging (BLI) to evaluate the efficiency of germ cell transplantation in vitro and in vivo in fertility preservation for infertile male mice.Design: Transgenic mouse model.Setting: University-based teaching hospital.Animal(s): Transgenic mice.Intervention(s): Busulfan was used to induce testicular failure in 3-week-old immature FVB/NJNarl wild-type recipient mice. At 8 weeks of age they received hemizygotic germinal cells from 3-week-old immature male FVB/N-Tg (PolII-luc) Ltc strain transgenic donor mice, transplanted into the seminiferous tubules.Main Outcome Measure(s): Isolated germinal cells were suspended in multiwell plates with the bioluminescent substrate d-luciferin in excess to quantify viable germ cells in vitro. Quantitatively in vivo BLI was applied to demonstrate the efficiency and success of transplantation and BLI of live pups born from wild-type in vivo.Result(s): Live birth pup of FVB/N-Tg (PolII-luc) Ltc transgenic mouse were born and imaged by bioluminescence after mating FVB/NJNarl female wild-type and male wild-type infertile recipient 4–6 months after transplantation of germinal cells of FVB/N-Tg (PolII-luc).Conclusion(s): The BLI could be applied successfully to this transgenic small animal model. It proved a useful tool for quantifying germ cells in vitro and for assessing the efficacy of germ cell transplantation in vivo.

Arachidonic acid regulation of the cytosolic phospholipase A2α/cyclooxygenase-2 pathway in mouse endometrial stromal cells

2012-02-27

Objective: To investigate the role of arachidonic acid (AA) in mouse endometrial stromal cells.Design: Experimental animal study.Setting: University research laboratory.Animal(s): Sexually mature female CD1-strain mice.Intervention(s): Primary culture of endometrial stromal cells.Main Outcome Measure(s): Western blot and real-time polymerase chain reaction for gene expression and/or phophorylation analysis. Luciferase assay for Cox-2 promoter analysis.Result(s): AA-derived prostaglandins play important roles during embryo implantation and decidualization. However, the function of AA itself in reproduction is largely unknown. In this study, exogenous AA stimulated cPLA2α phosphorylation and COX-2 expression, mainly through ERK1/2 in mouse endometrial stromal cells, and p38 inhibitor modestly inhibited cPLA2α phosphorylation induced by AA. The induction of COX-2 by AA was diminished by short interfering RNA against C/EBPβ and inhibitory C/EBPβ (LIP). C/EBPβ binding site at −872–−864 of Cox-2 promoter contributes to Cox-2 promoter activation induced by C/EBPβ transfection. The expression of C/EBPβ protein induced by AA was inhibited by p38 inhibitor, and the phosphorylation of C/EBPβ induced by AA was inhibited by p38 inhibitor and ERK1/2 inhibitor. A nonmetabolized analogue of AA (ETYA) also enhanced cPLA2α phosphorylation and COX-2 expression. The activation of cPLA2α/COX-2 by AA was not inhibited by COX inhibitor indomethacin.Conclusion(s): AA can induce cPLA2α/COX-2 pathway activation in mouse endometrial stromal cells.

Localization of B4GALNT2 and its role in mouse embryo attachment

Pei-Tzu Li, Chi-Jr Liao, Lung-Chi Yu, Wen-Guey Wu, Sin Tak Chu — 2012-03-09

Objective: To investigate the location of β-1,4-N-acetylgalactosaminyltransferase II (B4GALNT2) and the involvement of this protein and Sda antigen in embryonic implantation.Design: Cell and animal study.Setting: University.Animal(s): Adult outbred Institute for Cancer Research mice.Intervention(s): B4GALNT2 antibody injected into the uteri of mice in early pregnancy; E3.5 blastocysts and pregnant uterine tissues were collected.Main Outcome Measure(s): Protein expression was detected by immunofluorescence staining and Western blotting. Embryo attachment was assayed via in vitro and in vivo embryo implantation models.Result(s): The b4galnt2 gene expression in the 293T cell line showed the protein localized in the plasma membrane. We confirmed that B4GALNT2 was localized on the surface of E3.5 blastocysts but was an intracellular component in uterine epithelia. Finally, anti-B4GALNT2 and lectins inhibition assays demonstrated the involvement of B4GALNT2 and Sda antigen in embryonic attachment in vitro and in vivo via the mouse system and human endometrial cell line (Ishikawa).Conclusion(s): B4GALNT2 expressed in the blastocyst may interact with a ligand on the endometrial surface, perhaps via Sda also, to permit embryo implantation. Our data suggest that B4GALNT2 and Sda antigen are essential for embryo implantation.

Analysis of the expression of neurokinin B, kisspeptin, and their cognate receptors NK3R and KISS1R in the human female genital tract

2012-03-15

Objective: To investigate the presence of neurokinin B (NKB)/NK3 receptor (NK3R) and kisspeptin/KISS1 receptor (KISS1R) messenger RNA (mRNA) and proteins throughout the human female genital tract.Design: In vitro study.Setting: Academic research laboratories and academic hospitals.Patient(s): Fifteen reproductive-age women and 16 postmenopausal women provided fresh samples of uterus, ovary, or oviduct, and 12 women provided archival samples of endometrium or oviduct.Intervention(s): Fresh and archival samples of uterus, ovary, and oviduct obtained from reproductive-age and postmenopausal women.Main Outcome Measure(s): Results of reverse-transcription polymerase chain reaction (RT-PCR) and immunohistochemistry to investigate the pattern of expression of NKB/NK3R and kisspeptin/KISS1R in target tissues.Result(s): Expression of the genes encoding NKB (TAC3) and NK3R (TACR3), and kisspeptin (KISS1) and its receptor (KISS1R) was found in the uterus, ovary, and oviduct. Both NKB and NK3R immunoreactivity was detected in the endometrium, the oviduct, and the ovary, with marked expression in endometrial and oviductal epithelial cells, where intense coexpression of kisspeptin and KISS1R was also detected. Positive staining for NKB and NK3R was found in the myometrium where, in contrast, kisspeptin and KISS1R were absent.Conclusion(s): NKB/NK3R and kisspeptin/KISS1R are present in female peripheral reproductive tissues with colocalization of both systems in some non-neuronal cell populations of the human female genital tract. Our findings are compatible with a potential modulatory role of NKB and kisspeptin at peripheral reproductive tissues.

Efficiency of slush nitrogen vitrification of human oocytes vitrified with or without cumulus cells in relation to survival rate and meiotic spindle competence

2012-03-09

Objective: To evaluate the efficiency of slush nitrogen vitrification of human oocytes with or without cumulus cells in terms of survival rate and maintenance of meiotic spindle.Design: Randomized, comparative study.Setting: Medical center.Patient(s): A total of 274 oocytes obtained from 46 couples undergoing infertility treatment.Intervention(s): Metaphase II oocytes were divided into groups A and B, vitrified with and without cumulus cells, respectively.Main Outcomes Measure(s): Survival rates and maintenance of meiotic spindle observed immediately after warming and 3 hours after incubation.Result(s): No statistically significant difference was detected between the two groups in terms of survival rate, but a significantly higher percentage of detectable spindle was observed in group B (completely denuded oocytes), either immediately after warming or 3 hours after incubation.Conclusion(s): Complete denudation of oocytes before slush nitrogen vitrification does not influence survival rates but positively affects oocyte meiotic spindle competence. These data support the hypothesis that cumulus cells during vitrification represent an obstacle to cryoprotectant penetration more than having a protective role for the oocyte.

Anti-ACTL7a antibodies: a cause of infertility

2012-03-05

Objective: To show that antibodies against ACTL7a, a spermatozoon-specific protein, may be a cause of immunologic infertility.Design: Determine the presence of anti-ACTL7a antibodies in infertile blood, raise antibodies against ACTL7a in rabbits, and demonstrate that the in vitro treatment of mouse spermatozoa with infertile sera markedly reduces their fertilizing capacity. Demonstrate that the active immunization of mice with ACTL7a protein reduces fertility.Setting: National Research Institute for Family Planning, Beijing, World Health Organization Collaboration Center of Human Reproduction, China.Animal(s): Rabbits, ICR mice.Intervention(s): None.Main Outcome Measure(s): Mass spectrometry, indirect immunostaining, spermatozoa agglutination test, and standard fertility assay.Result(s): The fertilizing potential of mouse spermatozoa was markedly reduced after in vitro treatment with ACTL7a antibody–containing serum from a vasectomized man. Active immunization with ACTL7a significantly reduced the fertility of mice. Anti-ACTL7a antibodies caused the agglutination of mouse and human spermatozoa in vitro. Furthermore, the antibodies were detected in the sera of additional vasectomized men.Conclusion(s): Anti-ACTL7a antibodies may cause infertility in mice because the in vitro treatment of mouse spermatozoa with ACTL7a antibody–containing serum markedly reduced the fertilizing potential of the spermatozoa. In addition, the active immunization of mice with ACTL7a resulted in significant reductions in fertility.

Administration of visfatin during superovulation improves developmental competency of oocytes and fertility potential in aged female mice

2012-03-16

Objective: To examine whether visfatin administration during superovulation improves ovarian response, developmental competence of oocytes, and fertility in aged female mice.Design: Controlled experimental study.Setting: University hospital.Animal(s): Two groups of differently aged C57BL female mice (6–11 and 26–31 weeks).Intervention(s): Female mice were coinjected intraperitoneally with 5 IU pregnant mare's serum gonadotropin (PMSG) and visfatin of various doses (0–500 ng/mL), followed by 5 IU human chorionic gonadotropin (hCG) injection 48 hours later. Then the mice were immediately mated with an individual male. After 18 hours zygotes were cultured, and expression of ovarian visfatin and vascular endothelial growth factor (VEGF) was examined. Potential pregnancies of visfatin-administered aged female mice were monitored for delivery of offspring.Main Outcome Measure(s): Number of zygotes retrieved, embryo developmental competency, fertility potential, ovarian visfatin and VEGF expression.Result(s): Ovarian visfatin expression was significantly decreased in the aged mice group compared with the young. Visfatin administration significantly increased embryo developmental rate and ovarian visfatin and VEGF expressions in the aged mice. Visfatin-administered aged mice delivered significantly higher numbers of offspring than controls.Conclusion(s): This study suggests that visfatin administration during superovulation plays an important role in regulating oocyte quality and can improve oocyte quality and fertility of aged female mice.

Circulating antimüllerian hormone levels in boys decline during early puberty and correlate with inhibin B

2012-03-12

Objective: To investigate peripheral levels of inhibin B and antimüllerian hormone (AMH) in boys during peripuberty and in patients with congenital hypogonadotropic hypogonadism (HH).Design: Randomized, placebo-controlled trial (peripubertal boys); and cross-sectional clinical study (males with HH).Setting: University central hospital.Patient(s): Twenty-eight peripubertal boys with idiopathic short stature (ISS), 19 males with Kallmann syndrome.Intervention(s): Letrozole (2.5 mg/day) or placebo in boys with ISS for 2 years.Main Outcome Measure(s): Longitudinal follow-up observation of serum AMH and its relationship with inhibin B during early puberty and the influence of high (letrozole-treated boys) and low (males with HH) gonadotropin exposure on circulating AMH.Result(s): In boys with ISS receiving placebo, the decrease in AMH levels and the increase in inhibin B levels were correlated. The serum AMH level had already declined before a clinically significant increase in testis volume or serum testosterone occurred. Letrozole did not appear to modulate the decline in AMH. The AMH levels were lower in boys and young adults with Kallmann syndrome and prepubertal testes (mean: 20.9 ± 4.7 ng/mL, n = 6) as compared with prepubertal ISS boys (102.3 ± 11.9 ng/mL).Conclusion(s): The gonadotropin-mediated early pubertal increase in inhibin B is tightly coupled to decrease in AMH levels and may reflect androgen-mediated differentiation of Sertoli cells. Profound gonadotropin deficiency is associated with low AMH levels, suggesting impaired development of the Sertoli cell population.

Chromosome transfer in mature oocytes

Masahito Tachibana, Michelle Sparman, Shoukhrat Mitalipov — 2012-05-01

Objective: To demonstrate step-by-step micromanipulation procedures required for transfer of spindle-chromosomal complexes between mature oocytes.Design: Video presentation of reproductive biology study.Setting: In vitro fertilization and embryo manipulation laboratory.Animal(s): Rhesus (Macaca mulatta) primates.Intervention(s): Transplantation of the genetic material between mammalian oocytes offers many opportunities to study various aspects of nuclear-cytoplasmic interactions during oogenesis, fertilization and embryo development. We demonstrate the feasibility of isolation and transfer of chromosomes between mature metaphase II (MII) primate oocyte. After fertilization, manipulated oocytes were capable of producing healthy offspring or embryonic stem cells.Main Outcome Measure(s): In this video, we show micromanipulation procedures required for isolation and transfer of spindle-chromosomal complexes between rhesus MII oocytes. In brief, the spindle is visualized using a polarized microscope and extracted into a membrane enclosed karyoplast. Karyoplasts are then reintroduced into an enucleated recipient oocyte (cytoplast, derived from an another female) by karyoplast-cytoplast membrane fusion.Result(s): Newly reconstructed oocytes consist of nuclear genetic material from one female and cytoplasmic components, including mitochondria and mitochondrial DNA from another.Conclusion(s): This video demonstrates the protocol developed for primate oocytes that successfully allowed of isolation and transfer of chromosomes between mature metaphase II (MII) oocytes. Potential clinical applications include mitochondrial gene replacement therapy to prevent transmission of mtDNA mutations and treatment of infertility caused by cytoplasmic defects in oocytes. Video is available at http://fertstertforum.com/2012974tachibana/.

Single port laparoscopy

Luiz Carvalho, Rebecca L. Flyckt, Pedro F. Escobar, Tommaso Falcone — 2012-05-01

Objective: To demonstrate the abdominal wall anatomy necessary to perform a single port laparoscopic procedure. Single port laparoscopic (SPL) surgery has reduced the number of sites required to perform laparoscopic surgery. However, the incision at the umbilicus is larger than conventional laparoscopic surgery.Design: Video presentation of clinical article. The video uses animation and surgical cases to demonstrate the relevant abdominal wall anatomy to establish surgical access for a single site or single port laparoscopy.Result(s): This video demonstrates the regional anatomy pertinent to the anterior abdominal wall, specifically of the umbilicus. The umbilicus is a focal point of fusion of the anterior abdominal wall muscles that allows entry into the peritoneal cavity. For this procedure there are 2 incisions possible, a small midline intra-umbilical one and an omega incision. The video demonstrates each technique. Introduction of a port into this single incision is demonstrated with 2 different trocar systems. These trocar systems show how the limitations of using a single site may be reduced.Conclusion(s): The abdominal wall anatomy is unique at the umbilicus and allows optimal placement of a single trocar to allow laparoscopic surgery. Video is available at http://fertstertforum.com/2012974caravalho/.

Quantification of mitochondrial abundance

Tuncer Cayci, Bulent Kurt, Emin Ozgur Akgul, Yasemin Gulcan Kurt — 2012-03-28

We read the article by Duran et al. titled “The association of reproductive senescence with mitochondrial quantity, function, and DNA integrity in human oocytes at different stages of maturation” with great interest. In that article, the authors aimed to investigate the contribution of mitochondrial aging to female reproductive senescence in humans. To test this hypothesis, an attempt was made to estimate the number of mitochondria by the quantification of mitochondrial (mt) DNA copy number, which is measured via real-time polymerase chain reaction. They also studied some other biochemical and molecular parameters. They found that the number of mitochondria in the oocytes from various maturational stages did not show any statistically significant differences. They hypothesized that reproductive aging may lead to decreased numbers of mitochondria in human oocytes, which are interpolated by using mtDNA copy number. In their discussion, they concluded that inherently imperfect ovarian reserve indicators, such as FSH level and total number of oocytes retrieved by controlled ovarian hyperstimulation, may also predict the number of mitochondria in individual human oocytes.

Reply of the Authors

Frank Joseph Castora, Eyup H. Duran — 2012-03-28

We thank Dr. Kurt and his colleagues for their interest in our paper and their observations about our assessment of the number of mitochondria in the individual oocytes we examined. We agree that this estimate of mitochondrial number based on real-time polymerase chain reaction (PCR) assays of mitochondrial (mt) DNA is imprecise. As noted by Dr. Kurt and colleagues, aside from any imprecision associated with the real-time PCR methodology, there is the uncertainty associated with the current accepted estimate of mtDNA molecules per mitochondrion in oocytes. Is it one? Is it two?

Inadequate description of an acupuncture treatment study for polycystic ovaries

Roy Moncayo, Helga Moncayo — 2012-03-26

In the February issue of the Journal, Stener-Victorin et al. have published an article that confuses concepts and methods of acupuncture. In the title of the publication, the authors refer to the effects of acupuncture; however, in the publication itself they refer to electroacupuncture. Electroacupuncture does not belong to the traditional treatment methods in Chinese medicine . Practitioners of acupuncture, such as ourselves, are confused by such title-line statements. In addition to this, traditional acupuncture works with clearly and exactly defined acupoints to allow reproducibility and comprehension of any kind of treatment . Stener-Victorin et al. fail to describe the points used in their therapeutic approach. They vaguely mention the use of acupoints, described as “inserted bilaterally in acupuncture points in abdominal muscles and in muscles below the knee” . Their publication does not provide any reproducible thoughts that adhere to acupuncture. In addition to this, the use of acupoints related to muscular structures can be seen as a defined interrelationship within the concept of the Five Elements , as we demonstrated in 2009 .

Reply of the Authors

Elisabet Stener-Victorin — 2012-03-26

It is unclear what concept confuses the readers of our recently published article reporting secondary analyses of a randomized controlled trial . Acupuncture (Latin acus, “needle”; punctura, “puncture”) is an integral part of traditional Chinese medicine, and over the last decade it has become established in Western medicine. Acupuncture also involves stimulation of the needles, which may be via manual manipulation, electrical stimulation, or by heating (moxa). It is unfortunate if the readers become confused because we do not indicate the type of stimulation in the title. However, type of stimulation, which is a combination of manual and electrical stimulation of the needles, is clearly described in the article, though not the exact points because they have already been published in the primary article referred to and in an experimental part of the trial . In the primary article, electroacupuncture is given in the title , which is already too long and therefore the main reason not to include “electro” in the present article owing to limited space. Importantly, exact points, stimulation (both manual and electrical), number of needles, depth of needle placement, treatment time, number of treatments, and such are reported in the primary article and briefly in the present article, all according to the Standards for Reporting Interventions in Clinical Trials of Acupuncture (STRICTA) . Furthermore, the acupuncture rationale in the present study is Western Medical Acupuncture according to STRICTA definition . The reader needs to check the references, and if there is any uncertainty they should contact the corresponding author. However, it seems that in this case it is rather the rationale that is a problem, and that is clearly stated in both the primary and secondary articles. It is striking also that the comment refers to textbooks rather than scientific literature. In a scientific article we need to stick to science and, most importantly today, to increase the quality of acupuncture research by following the CONSORT and STRICTA guidelines, as done in the present study .

Comment on, “Is there a place for corifollitropin alfa in IVF/ICSI cycles? A systematic review and meta-analysis”

Bernadette Mannaerts, Pierre Verweij, Keith Gordon — 2012-03-28

As the sponsor of all four trials included in a recent systematic review and meta-analysis of corifollitropin alfa, we reviewed the manuscript authored by Youssef et al. with great interest. This combined analysis includes two phase II dose-finding trials and two phase III randomized controlled trials.

Reply of the Authors

2012-03-28

We read the letter by Mannaerts et al. regarding our meta-analysis on corifollitropin alfa with interest. We welcome this opportunity to apologize for the typing error in the number needed to harm (NNH). The correct NNH is indeed almost 100.

Vitrification carriers and European regulation

Lodovico Parmegiani, Graciela Estela Cognigni, Marco Filicori — 2012-03-22

We read with interest the recent article by Valbuena et al. comparing the efficiency of Cryotip versus Cryotop for blastomere vitrification. The authors concluded that it is preferable to preserve individual human blastomeres using Cryotip, which is a closed system. Furthermore, they suggest that this has the advantage of complying with European Union directives.

Reply of the Authors

Diana Valbuena, Carlos Simon — 2012-03-22

We have read the letter to the editor by Parmegiani et al. entitled “Vitrification Carriers and European Regulation” referring to our recent article . In our work, we compared open versus closed systems for mice and human blastomere vitrification and concluded that the closed system is more efficient in terms of blastomere survival and division.

Acupuncture for IVF: do not let needles stand in the way of empathy

Jean-Luc Mommaerts, Dirk Devroey — 2012-03-26

Zheng et al. claim in their meta-analysis that there is no significant difference between acupuncture and placebo when it comes to live birth rates among women undergoing in vitro fertilization because the Streitberger procedure, in which needles do not enter the skin and which was used as placebo acupuncture, appears to be more effective than acupuncture itself. This might be interpreted as a recommendation to choose the Streitberger procedure over acupuncture as the active treatment. The authors seem to agree, saying “the harmful reaction produced by real acupuncture can be avoided by this noninvasive stimulation.”

Hormonal therapy for male sexual dysfunction

Edward D. Kim — 2012-04-01

Over the last decade, there has been a significantly increased interest in the use of testosterone and other hormonal therapies for the aging male. For many baby-boomer generation men, the hope is that these hormonal therapies will improve their sexual function and treat their sexual dysfunction. However, despite advances in the understanding of sexual physiology, the field of sexual endocrinology is still incompletely understood.

Editorial Board

2012-05-01

Fertility and Sterility

Recurrent implantation failure: current update and clinical approach to an ongoing challenge

Recurrent implantation failure: gamete and embryo factors

Defective endometrial receptivity

Recurrent IVF failure: other factors

Repeated implantation failure: clinical approach

A persistent misperception: assisted reproductive technology can reverse the “aged biological clock”

A good meta-analysis is hard to find

The incidence and effect of bacteriospermia and elevated seminal leukocytes on semen parameters

Electroejaculation combined with intracytoplasmic sperm injection in patients with psychogenic anejaculation yields comparable results to patients with spinal cord injuries

A platform for evaluating sperm RNA biomarkers: dysplasia of the fibrous sheath—testing the concept

DNA integrity is maintained after freeze-drying of human spermatozoa

Emergency testicular sperm extraction after scrotal trauma in a patient with a history of contralateral orchiopexy for cryptorchidism: case report and review of the literature

An investigation into the relationship between the metabolic profile of follicular fluid, oocyte developmental potential, and implantation outcome

Effects of laser polar-body biopsy on embryo quality

What is the optimal follicular size before triggering ovulation in intrauterine insemination cycles with clomiphene citrate or letrozole? An analysis of 988 cycles

How members of the Society for Reproductive Endocrinology and Infertility and Society of Reproductive Surgeons evaluate, define, and manage hydrosalpinges

Oocyte slow freezing using a 0.2–0.3 M sucrose concentration protocol: is it really the time to trash the cryopreservation machine?

The use of recombinant luteinizing hormone in patients undergoing assisted reproductive techniques with advanced reproductive age: a systematic review and meta-analysis

Why does the fallopian tube fail in ectopic pregnancy? The role of activins, inducible nitric oxide synthase, and MUC1 in ectopic implantation

Genetic association study of polymorphisms FOXP3 and FCRL3 in women with endometriosis

Increased expression of macrophage colony–stimulating factor and its receptor in patients with endometriosis

A prospective cohort study of physical activity and time to pregnancy

Glutathione transferase polymorphisms and risk of endometriosis associated with polychlorinated biphenyls exposure in Italian women: a gene–environment interaction

Orthotopic grafting of cryopreserved prepubertal testicular tissue: in search of a simple yet effective cryopreservation protocol

Microarray expression profiling in adhesion and normal peritoneal tissues

A novel luteinizing hormone/chorionic gonadotropin receptor mutation associated with amenorrhea, low oocyte yield, and recurrent pregnancy loss

Assessment of FSHR variants and antimüllerian hormone in infertility patients with a reduced ovarian response to gonadotropin stimulation

Viability and function of the cryopreserved whole rat ovary: comparison between slow-freezing and vitrification

The impairment of reproduction in db/db mice is not mediated by intraovarian defective leptin signaling

Bioluminescence imaging as a tool to evaluate germ cells in vitro and transplantation in vivo as fertility preservation of prepubertal male mice

Arachidonic acid regulation of the cytosolic phospholipase A2α/cyclooxygenase-2 pathway in mouse endometrial stromal cells

Localization of B4GALNT2 and its role in mouse embryo attachment

Analysis of the expression of neurokinin B, kisspeptin, and their cognate receptors NK3R and KISS1R in the human female genital tract

Efficiency of slush nitrogen vitrification of human oocytes vitrified with or without cumulus cells in relation to survival rate and meiotic spindle competence

Anti-ACTL7a antibodies: a cause of infertility

Administration of visfatin during superovulation improves developmental competency of oocytes and fertility potential in aged female mice

Circulating antimüllerian hormone levels in boys decline during early puberty and correlate with inhibin B

Chromosome transfer in mature oocytes

Single port laparoscopy

Quantification of mitochondrial abundance

Reply of the Authors

Inadequate description of an acupuncture treatment study for polycystic ovaries

Reply of the Authors

Comment on, “Is there a place for corifollitropin alfa in IVF/ICSI cycles? A systematic review and meta-analysis”

Reply of the Authors

Vitrification carriers and European regulation

Reply of the Authors

Acupuncture for IVF: do not let needles stand in the way of empathy

Hormonal therapy for male sexual dysfunction

Editorial Board

Table of Contents