Recurrent In Vitro Fertilization Failure Research Articles

The Association of NKp46-Positive uNK Cells With a Higher Risk of Recurrent Miscarriage and IVF Failure

Background: Uterine natural killer (uNK) cells have a significant impact on pregnancy and related complications. Given the importance of receptors in the activity of uNK cells, the present study aimed to determine the number of uNK cells and NKp46 (one of the most important NK cell-activating receptors) expression in the endometrium of women with recurrent miscarriage (RM) or a history of in vitro fertilization (IVF) failure. Materials and Methods: This case-control study was performed on 48 participants, including 16 healthy controls, 27 cases with RM, and 5 cases with repeated implantation failure (RIF) during the mid-luteal phase according to a standardized diagnostic protocol. All participants were assessed using transvaginal ultrasound to determine embryo survival rate and confirm gestational age. Endometrial specimens were collected and subjected to immunohistochemistry (IHC) staining using an anti-human NKp46 antibody expressed by uNK cells. Results: A significantly higher number of cells positive for NKp46 was obtained among two groups of cases versus healthy subjects (patients: 1.46±0.78, controls: 0.82±62, P=0.006), and the number of CD56+cells was significantly higher in patients than in controls (patients: 18.14±7.14, controls: 11.71±6.17, P=0.003). Additionally, there was not a significant difference in the frequency ratio of NKp46+NK cell subset to CD56+uNK cells between the patients (P=0.59) and control healthy group. Conclusion: The increase in the number of uterine NK cells and their cytotoxic activity during implantation and early pregnancy, possibly resulting from an excessive expression of inflammatory cytokines, confirms a significant association between uNK cell activity and a higher risk of RM and RIF. Therefore, immunomodulatory treatments may benefit these patients.

P-329 Oocyte donation outcomes in endometriosis patients with multiple IVF failures

Abstract Study question What are the reproductive outcomes and the prognostic factors of live birth rates in endometriosis patients referred to oocyte donation after multiple in vitro fertilization(IVF)-failures? Summary answer In this specific population, oocyte donation appears to be a viable option to optimize the reproductive outcomes, as the cumulative live birth rate reaches 63.2%. What is known already Assisted Reproductive Technology(ART) is currently recognized as one of the main therapeutic options to manage endometriosis-related infertility, often resulting in satisfactory outcome. A burning issue in this field remains how to deal with patients undergoing recurrent ART failures. As there appear to be two main factors contributing to the failures, i.e., a deleterious effect of endometriosis on embryo implantation and altered oocyte and embryo quality, two competing therapeutic strategies can be considered:(i) endometriosis surgery and (ii) oocyte donation. However, no data has been published to date in the specific population of endometriosis patients with previous IVF failures, undergoing oocyte donation. Study design, size, duration Observational cohort study including fifty-seven women with endometriosis-related infertility and multiple IVF failures (≥ 2 failed IVF cycles) referred to oocyte donation who consulted at our institution between January 2013 and June 2022. Participants/materials, setting, methods Endometriosis was diagnosed based on published imaging criteria using transvaginal sonography and magnetic resonance imaging and confirmed histologically in women who had a history of surgery for endometriosis. The main outcomes measured were the clinical pregnancy rate (CPR) and the cumulative live birth rate (CLBR). We compared the characteristics of women who had a live birth versus those who did not using univariate and multivariate analysis to identify determinant factors of fertility outcome. Main results and the role of chance Fifty-seven patients underwent 90 oocyte donation cycles, after 244 failed autologous IVF cycles. The mean age of the population was 36.8 ± 3.3 years, with a mean infertility duration of 3.6 ± 2.2 years, and a mean number of autologous IVF/ICSI cycles of 4.4 ± 2.3 cycles per patient. Three patients (5.3%) had superficial endometriosis, 2 patients (3.5%) had ovarian endometriomas, and 52 patients (91.2%) had deep infiltrating endometriosis, among which 30 patients (57.7%) had bowel lesions. Thirty patients (52.6%) had associated adenomyosis. Overall, the CPR and the CLBR per patient were 49/57 (85.9%) and 36/57 (63.2%), respectively. After multivariate analysis, only being nulligravida (p = 0.002) remained an independent negative predictive factor of the live birth rate. Limitations, reasons for caution In 35.1% of patients there was no surgical/histological proof of endometriosis. However, for all the study participants, endometriosis was diagnosed and staged using patient interviews, clinical examination, and imaging techniques to assess for endometriosis phenotypes, which is in line with the structured diagnostic process acknowledged as being appropriate for endometriosis. Wider implications of the findings In this specific population, poor embryo quality may be the main limiting factor of ART success rates, rather than impaired endometrial receptivity. These data are reason for clinicians not to systematically refer infertile endometriosis patients with recurrent IVF failures to surgery. Trial registration number not applicable

Endometrial Staining of CD56 (Uterine Natural Killer), BCL-6, and CD138 (Plasma Cells) Improve Diagnosis and Clinical Pregnancy Outcomes in Unexplained Infertility and Recurrent IVF Failures: Standardization of Diagnosis with Digital Pathology.

In women with unexplained infertility (UI) and recurrent in vitro fertilization (IVF) failures, the etiology is often unclear. Endometrial immune perturbations and the use of immune markers associated with these dysregulations are of great interest in the diagnosis and treatment of UI. However, reliable biomarkers and standardized quantification methods are lacking. Here, to address endometrial immune dysregulation in UI patients with recurrent IVF failures, we performed endometrial tissue sampling and immunostaining of CD56 (uNK), CD138, and BCL-6. Of these cases, 57.9% had positive CD56 in the endometrial stroma, while 46.1% had positive BCL-6 in the glandular epithelium, and 14.5% of the cases were found to be positive for CD138. Combined staining rates were 60.5%, 68.4%, and 71.05% for (CD56 or BCL-6), (CD56 or CD138), and (CD56, BCL-6, or CD138), respectively. There was a significant correlation between CD56 and BCL-6 positivity, while CD138 positivity was an independent parameter. After the recommended targeted therapy, pregnancy rates were found to increase from 58.5% to 61.6% and 73.8% in CD56-positive, (CD56- or BCL-6-positive), and (CD56-, BCL-6-, or CD138-positive) cases, respectively. Notably, a retrospective evaluation of digital pathology and light microscopy results showed a significant correlation. This study suggests that the examination of CD56, BCL-6, and CD138 in the same endometrial sample may be an effective method in determining the etiology of UI and reaching an early diagnosis and treatment options. Moreover, digital pathology can be used in the evaluation of CD56 and BCL-6 to provide objective, rapid, and reliable results.

Relation of PLCζ Gene with Sperm and Oocyte Parameters in Infertility Treatment based on Assisted Reproductive Technologies: Review

Introduction: As the world's population increases, the incidence of infertility is increasing, and infertility is now considered to affect 8–12% of couples worldwide. Assisted reproductive technology (ART) such as in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) is revolutionizing the field of reproductive medicine and strives continuously to provide infertile patients with access to appropriate and efficient treatments. However, some causes leading to total/low fertilization rate in ART have been addressed. Phospholipase C zeta (PLCζ) is the main sperm-specific factor responsible for triggering oocyte activation (OA). Therefore, PLCζ abnormalities, including low and absent expression as well as its mutation has been described as one of the fertility rate inducers resulted from the oocyte activation deficiency and sperm parameters abnormalities and failed IVF and ICSI cycles. In this review article we have aimed to focus on the latest information of the PLCζ published data in the Web of Science, Scopus, Science Direct and PubMed databases and resulted in the failed IVF and ICSI and recurrent ICSI and IVF failure. In addition the investigations on PLCζ gene and its coded protein in the animals have been also reported.
 Conclusion:.Studies have shown that the change in PLCζ expression and its mutations can cause a decrease in the fertilization rate due to egg activation defects and abnormalities in sperm parameters and failure in ART.

Comparison of Adjunctive Use of G-CSF Vs Autologous PRP in IVF Patients with a Refractory Thin Endometrium: A Retrospective Record Review

Background: In Vitro Fertilization (IVF) is an important treatment option in the management of couples with infertility. Sadly, however, failure to achieve a pregnancy through IVF is not uncommon. Amongst the many causes of IVF failure, implantation failure has emerged as one of the more common and important factors. The refractory thin endometrium as a cause of recurrent IVF failure has been well documented. The use of either Granulocyte Colony Stimulating Factor (G-CSF) or autologous Platelet Rich Plasma (PRP) has emerged as potential adjunctive treatments that may mitigate the rate of implantation failure; however, no conclusive evidence exists to favour the use of one over the other. Objective: To compare the measured change in endometrial thickness and pregnancy rates in patients with Recurrent implantation Failure (RIF) and/or thin endometrium following the intrauterine administration of either G-CSF or autologous PRP. In addition, to compare the pregnancy rates in patients with fluid in the endometrial cavity who underwent either therapy. Design: A retrospective analysis was conducted on patients who underwent endometrial therapy (either G-CSF or PRP) between January and June 2020. The measured change in endometrial thickness and the clinical pregnancy outcome of the two groups were compared. Subjects: 36 patients with a mean age of 40.36 years met the inclusion criteria of the study. 20 received autologous intrauterine PRP treatment and 16 received intrauterine G-CSF treatment. Both groups were well matched for age, pre-intervention endometrial thickness and embryo quality. Intervention: Administration of G-CSF (One ampoule Neupogen® (filgrastim)) or PRP (1ml) into the uterine cavity transcervical 48 hours prior to embryo transfer. Main Outcome Measures: The change in endometrial thickness measured 48 hours prior to embryo transfer and at the time of embryo transfer (ET) was compared. Positive clinical pregnancy outcome was determined by a positive serum B-HCG test 10 days post insertion. A statistically significant difference was set at p=0.05. Results: There was a statistically significant difference in endometrial expansion post intervention in both the G-CSF and PRP groups. However, the difference between the two groups did not reach statistical significance (p=0.077). Additionally, the collective pregnancy rate of the total study population was 44.4% (16 of 36), a significant increase over the expected pregnancy rate in the published literature [1,2]. Of the positive pregnancies, 9 (56.25%) were in the autologous PRP group and 7 (43.75%) in the G-CSF group. This difference was, however, determined not to be statistically significant (p=0.603). Conclusion: Both G-CSF and PRP are effective interventions in the management of the thin refractory endometrium. Both result in significant endometrial expansion and increased pregnancy rates. Despite a marginally higher endometrial response and pregnancy rate in the PRP group, the differences in these metrics between the two groups were not statistically significant.

Validation of the existence of genuine Empty follicle syndrome, versus false empty follicle syndrome to make definitive decisions in cases where recurrent IVF failure encountered secondary to absence of oocytes on ovum pick up-a short communication

Aim: Worldwide a big altercation exists with regards to the actual existence of the term “ Empty follicle syndrome’’(EFS), so much so that certain big authorities in the field have been believing that true EFS does not exist. Basically EFS is a syndrome when no functionally intact oocyte get retrieved when attempting an oocyte pick up (OPU) for a successful in vitro fertilization (IVF), however such patients encounter recurrent IVF failures. Since it is has become a big problem for the treating reproductive endocrinologist, besides the patient encountering recurrent IVF failures, it has become essential to differentiate the true EFS alias genuine Empty follicle syndrome (gEFS) from what is labeled today as the false empty follicle syndrome (fEFS). In view of the recently documented presence of mutations, gEFS got verified and appears to silence this biggest conflict that arose secondary to the existence of a false empty follicle syndrome(fEFS), where one could manage to get successful IVF outcomes subsequent to repeated hCG injections/ gonadotropin releasing hormone (GnRH) agonist in addition to pregnancy, with lot of clinicians believing there is no true term like EFS. Methods: Recently Yang et al., performed a study In tertiary a university based reproductive center in China that was comprised of a big cohort of patients that presented with gEFS. Genetic evaluation was conducted on 35 non correlated infertile patients who went through 16 failed IVF cycles in addition to oocyte degeneration, besides the subjects got a diagnosis of possessing a particular kind of EFS- cumulus oocytes complexes (COC’s) but possessed oocytes that were undergoing degeneration, with the utilization of whole –exome sequencing along with targeted Sanger sequencing. Results: Yang et al., found 22 innovative genetic variant of zona pellucida (ZP), genes in 18 subjects, that were inclusive of 20 variants in ZP 1 gene, 2 in ZP 2 gene in addition to 1 recurring variant in ZP3 gene that had been earlier documented. The homogenous /compound heterogenous ZP 1 mutations were inherited in an autosomal recessive manner, while the heterogenous variants of ZP 2 as well as ZP3 genes possessed an autosomal dominant manner of inheritance. Conclusions: These mutations were anticipated to be harmful in silico along with got further experimentally corroborated to be functionally null dependent on their ectopic expression in vitro. Thus with this further evidence that has been recently provided with regards to the existence of genuine Empty follicle syndrome (gEFS), it is significant for the youngsters to realize if they encounter similar cases after trial of rescue hCG injections / GnRH) agonist, not to further keep waiting, but evaluate further with regards to the existence of mutations for Zona Pellucida (ZP), ZP 1, ZP2 as well as ZP3 genes mutations, or LH/ chorionic gonadotropins receptor (LHCGR) gene mutation without subjecting the patient to repeated IVF, with her psychological as well as financial health in mind

In vitro Fertilization Procedures with Embryo Transfer and Their Association with Thrombophilia, Thrombosis and Early Antithrombotic Treatments.

In vitro fertilization (IVF) procedures have been frequently associated with antithrombotic treatments, in particular, to aspirin or low-molecular-weight heparin (LMWH). The rationale of this treatment is based on the increase of thrombotic risk occurring in this clinical context. Indeed, both prothrombotic changes of coagulation parameters specifically related to IVF procedures as well as the presence of potential thrombophilic alterations may concur to increase the risk in these women. Furthermore, the presence of thrombophilia has been suggested as a potential cause of recurrent IVF failures. Therefore, antithrombotic treatments have been historically planned to prevent thrombotic disorders during pharmacological ovarian stimulation and/or to increase a successful rate of pregnancy and live births after IVF with embryo transfer. However, up to date, the role of inherited and\\or acquired thrombophilia is still debated as well as a univocal therapeutic approach is lacking in women with infertility. The administration of antithrombotic drugs differs in several studies and even the dosages of aspirin and\\or low-molecular-weight heparin are different. This review focuses on underlining current evidence on the role of thrombophilia and thromboprophylaxis in women selected for IVF with embryo transfer.

Recurrent implantation failure is associated with increased levels of fetuin-A/alpha2-Heremans-Schmid-glycoprotein

ObjectiveRecurrent implantation failure is defined as the absence of implantation, after two or three consecutive cycles of in-vitro fertilization (IVF), intracytoplasmic sperm injection or frozen embryo replacement. Human fetuin-A/alpha2-Heremans-Schmid-glycoprotein is a plasma protein secreted by the liver that modulates insulin action in adipocytes. Increased fetuin-A promotes adipocyte dysfunction which results in decreased adiponectin and increased fatty acids and inflammatory cytokines. Fatty acids and inflammatory cytokines were previously reported in implantation failure. Also, fetuin-A inhibits receptor tyrosine kinase activity in trophoblast growth factors which decrease trophoblast viability and invasion. In this study, we aimed to find the association between fetuin-A and implantation failure. Study designA total of 78 women were included in this case-control study. Serum fetuin-A concentrations were measured in 42 women with recurrent IVF failure and 36 healthy women with regular cycles. ResultsThe mean serum fetuin-A levels of implantation failure and control women were 257.77 ± 32.18 and 219.59 ± 48.86 respectively with a p-value <0.001 (independent samples t-test). Our results showed a statistically significant difference between serum fetuin-A levels of implantation failure women and controls. ConclusionSo far reasons for implantation failure are only partially understood. The current study reveals the association between implantation failure and fetuin-A. Further studies with large population sizes are needed to investigate whether fetuin-A can be used as a marker before controlled ovarian stimulation began or regulation of fetuin-A levels with treatment or lifestyle interventions can improve implantation success.

Laparoscopic surgery: Any role in patients with unexplained infertility and failed in vitro fertilization cycles?

Patients who undergo several in-vitro fertilization (IVF) treatment cycles and fail to conceive present a frustrating problem to the clinician. When 1 cycle of IVF treatment fails, should we offer the couples to choose additional cycle of IVF instead of evaluation of the potential peritoneal factor? In cases of otherwise unexplained infertility, the investigation cannot be considered to be complete until laparoscopy has been performed. The aim of the study is to investigate the fertility outcome of laparoscopic treatment in infertile women with repeated IVF failures.This is a retrospective case-control study conducted in a tertiary care, academic teaching hospital from January 2012 to December 2015. Patients recruited in this study were classified into 2 groups. Study group (n = 45) were offered laparoscopy for evaluation of infertility, control group (n = 45) elected to proceed to IVF without laparoscopy. Diagnostic laparoscopy and subsequent excision of suspected endometriotic lesions, lysis of adhesion and treatment of tubal pathology were performed when indicated.Forty-four (97.8%) patients in study had pelvic pathologies and the treatment was performed at the same time. Twenty-four patients in study group conceived including 16 patients conceived spontaneously and 14 patients conceived with additional IVF following laparoscopy management. There was a significant difference in the ongoing pregnancy rates between patients conceived through IVF in study group and control group (41.9% vs 19.6%, P < .05).Laparoscopy in women with normal hysterosalpingography but recurrent IVF failures can detect unrecognized pelvic pathologies. Laparoscopy evaluation prior to additional cycle of IVF seems to improve the subsequent pregnancy rate.

Prevalence/Incidence of Hereditary and Acquired Thrombophilia Markers among Egyptian Females with Recurrent Pregnancy Loss or IVF Failure

Background: The largest percentage of failed in vitro fertilization (IVF) cycles, are due to lack of implantation. As hereditary and acquired thrombophilia can cause in placentation failure, it may have a role in recurrent IVF failure. Objective: Aim of this case-control study was to determine the most prevalent types of hereditary and acquired thrombophilia in women with recurrent IVF failures. Materials and Methods: Case group comprised 2466 women, with a history of recurrent IVF failure. Control group was comprised of 531 men (Most prevalent DVT gender presented in our Clinics) suffering from Active DVT (To correlate the impact those factors IVF failure with an active thrombotic effect among the control group). All participants were assessed for the presence of inherited thrombophilias including: Factor V Leiden, methyl tetrahydrofolate reductase (MTHFR) mutation, prothrombin mutation, plasminogen activator inhibitor-1 (PAI-1) mutation, Factor XIII, ACE, HPA1, and APO E and the homocystein level, protein S and C deficiency, antithrombin III (AT-III) deficiency, lupus anti-coagulant, NK markers and anticardiolipin. Positive results are compared as regard most prevalent combinations. Results: Having at least one thrombophilia known as a risk factor for recurrent IVF failure (95% CI=1.74-5.70, OR=3.15, p=0.00). Mutation of factor V Leiden (95% CI=1.26-10.27, OR=3.06, P=0.01) and homozygote form of MTHFR mutation (95% CI=1.55-97.86, OR=12.33, p=0.05) were also risk factors for recurrent IVF failure. However, we could not find significant difference in other inherited thrombophilia’s. Conclusion: Inherited thrombophilia is as prevalent in women with recurrent IVF failure compared men with multiple DVT. Homozygous APO E and ACE as well as heterozygous ACE, MTHFR A and PAI contribute as a major factor in patients with recurrent failure.

Endometrial cancer in a woman undergoing hysteroscopy for recurrent IVF failure

BackgroundHysteroscopy, despite being the undisputed gold standard for the examination of the uterine cavity, is controversial as a routine procedure in infertile women. However, benign intrauterine conditions are common in women suffering repeated in vitro fertilization (IVF) failure, and growing evidence suggests a unique diagnostic and therapeutic role for hysteroscopy. Endometrial malignancy, on the contrary, is unreported by large published series of women with repeated IVF failures undergoing hysteroscopy, and its impact on fertility, for obvious reasons, has not been studied.ResultsAn unsuspected endometrial cancer was diagnosed in an asymptomatic 38-year-old woman undergoing hysteroscopy because of several repeated failures of in vitro fertilization and embryo transfer.ConclusionsEndometrial cancer can be found at hysteroscopy in young women with repeated IVF failures. The possibility of repeatedly unsuccessful fertility treatments should be taken into account when counseling infertile women about conservative treatment of endometrial cancer.

Endometrial injury, the quality of embryos, and blastocyst transfer are the most important prognostic factors for in vitro fertilization success after previous repeated unsuccessful attempts

PurposeThe purpose of this study was to find out the most important prognostic factors for achieving a pregnancy after in vitro fertilization (IVF) in women with history of repeated unsuccessful IVF attempts.MethodsWe analyzed factors affecting pregnancy rate in a retrospective study including 429 IVF/ICSI cycles performed in women younger than 40 years with at least three previous consecutive failed IVF/ICSI attempts.ResultsClinical pregnancy was observed in 140/429 (32.6%) cycles. Clinical pregnancy rate (CPR) was significantly higher in cycles with LEI compared to cycles without LEI before embryo transfer (44.4 vs 26.54%, p = 0.007). The CPR was also higher in cycles with day 5 blastocyst- compared with day 3 cleavage-stage embryo transfers (45.51 vs 26.54%, p < 0.001). In multivariate logistic regression model, only transfer of at least one good quality embryo (OR = 4.32, 95% CI 2.41–7.73), local endometrial injury (OR = 1.73, 95% CI 1.02–2.92), and transfer on day 5 (OR = 3.02, 95% CI 1.53–5.94) remained important independent prognostic factors for clinical pregnancy.ConclusionsThese results suggest that hysteroscopy with local injury to the endometrium prior to ovarian stimulation for IVF/ICSI can improve implantation and pregnancy rates in women experiencing recurrent IVF failure. However, large studies are needed to confirm these findings.

Pre-implantation genetic screening among women experiencing recurrent failure of in vitro fertilization.

To determine the efficiency of pre-implantation genetic screening (PGS) among women scheduled to undergo intracytoplasmic sperm injection who had experienced recurrent in vitro fertilization (IVF) failure. The present retrospective cohort study reviewed the medical records of consecutive women who had experienced recurrent IVF failure and had presented at a private IVF facility in Trabzon Province, Turkey, to undergo intracytoplasmic sperm injection between May 1, 2012, and December 31, 2014. Patient data and perinatal outcomes were compared between patients who underwent PGS and those who did not. There were 88 patients included in the study; 43 patients had undergone PGS and 45 had declined to do so. No differences were detected in the clinical pregnancy rate (P=0.846), spontaneous abortion rate (P=0.416), number of perinatal deaths (P=0.162), or the number of live deliveries (P=0.188) between the groups of patients. The pregnancies included in the study resulted in 25 neonates being delivered; 24 had normal karyotypes, and one neonate from the control group had a karyotype of 46, XX, 9ph. Among the 19 embryos that were not transferred, the most frequently encountered chromosomal anomalies were diploidy, monosomy X, and 2N/N/4N mosaicism, detected in 7 (37%), 2 (11%), and 2 (11%) embryos, respectively. PGS had no effect on perinatal outcomes among women experiencing recurrent IVF failure.

Recurrent IVF failure is associated with elevated progesterone on the day of hCG administration

ObjectiveDuring in vitro fertilization (IVF) treatment, elevated progesterone on the day of human chorionic gonadotrophin (hCG) administration has been reported to be associated with a reduced chance of live birth. It is not known, however, if the relationship is casual or causal. In the latter situation, one would expect the incidence of elevated progesterone on the day of hCG administration to increase with the number of IVF/embryo transfer (ET) failures. The aim of this study was to investigate if the frequency of elevated progesterone on the day of hCG administration is related to the number of IVF failures. Study designThis retrospective, observational, cohort study included a consecutive series of 6673 IVF cycles. Subjects were categorized into one of three groups: Group I, no previous IVF/ET treatment; Group II, one previous IVF/ET treatment failure; or Group III, two or more previous IVF/ET treatment failures. The main outcome measure was the proportion of cycles with elevated progesterone (>6nmol/l) on the day of hCG administration. ResultsAfter adjusting for age, oestradiol level on the day of hCG administration and number of oocytes retrieved, the proportion of women with elevated progesterone on the day of hCG administration remained significantly different between the three groups: Group I, 16.8%; Group II, 31.7%; and Group III, 39.7% (p<0.001). ConclusionElevated progesterone on the day of hCG administration is more likely in women with recurrent IVF failure. Women with two or more IVF failures are twice as likely to have elevated progesterone on the day of hCG administration as women undergoing their first IVF cycle.

Preimplantation genetic diagnosis in Saudi Arabia.

Preimplantation genetic diagnosis (PGD) testing is the practice of obtaining a cellular biopsy sample from a developing human oocyte or embryo, acquired via a cycle of in vitro fertilization (IVF); evaluating the genetic composition of this sample; and using this information to determine which embryos will be optimal for subsequent uterine transfer. PGD has become an increasingly useful adjunct to IVF procedures. The ability to provide couples who are known carriers of genetic abnormalities the opportunity to deliver healthy babies has opened a new frontier in reproductive medicine. The purpose of the PGD is enables us to choose which embryos will be implanted into the mother. In the present study 137 families who had undergone IVF at Habib Medical Centre, were enrolled for the PGD analysis. The couple visited the clinic for the sex selection, recurrent fetal loss and with the recurrent IVF failure. 802 embryos were tested by the biopsy method and 512 are found to be normal and 290 were abnormal embryos. In this study only 24% of the embryos were transferred and the remaining was not transferred because of the abnormalities or undesired sex of the embryos. The structural and numerical abnormalities were found to be 16.8%.

Does the prevalence of individual chromosomal aneuploidy vary with respect to day 3 embryo morphology?

Preimplantation genetic diagnosis (PGD) has been increasingly utilized in conjunction with embryo morphology grading to select embryos for transfer. Our group previously reported that High Implantation Potential (HIP) embryos are more likely to develop to the blastocyst stage, but a significant proportion of those are aneuploid. We aim to compare proportions of aneuploidy among individual chromosomes in HIP and non-HIP embryos that develop to the blastocyst stage by day 5. Retrospective analysis. We identified all women who underwent in vitro fertilization (IVF) with their first PGD cycle for recurrent miscarriage, recurrent IVF failure, advanced maternal age, or other indications between January 1 and December 31, 2006. Embryos were scored as HIP if they had ≥4 cells on day 2 and ≥7 cells on day 3, with <20% fragmentation and no multinucleation. Fluorescence in situ hybridization was utilized to analyze 9 or 12 chromosomes. P values were adjusted to account for lack of independence among embryos from the same woman. A total of 364 embryos from 88 cycles in 88 patients were analyzed. Non-HIP embryos were more likely to be aneuploid for chromosomes XY, 8, 15, 16, 18, and 22 compared to HIP embryos, with similar trends for chromosome 14 and 17. There were no significant differences between HIP and Non-HIP embryos in the aneuploidy rates of chromosomes 13, 20, and 21. TableAneuploidy Prevalence by ChromosomeChromosomeHIPNon-HIPPXY18/205 (9%)25/148 (17%)0.04811/63 (17%)12/34 (35%)0.011343/204 (21%)36/149 (24%)0.491417/68 (25%)15/35 (43%)0.091538/198 (19%)48/143 (34%)0.0011637/200 (19%)40/149 (27%)0.0471720/200 (10%)26/149 (17%)0.051838/204 (19%)45/149 (30%)0.012016/63 (25%)9/34 (26%)0.902146/206 (22%)34/150 (23%)0.942236/199 (18%)39/147 (27%)0.02No Nucleus4/210 (2%)4/154 (3%)0.64 Open table in a new tab Aneuploidy is common in embryos that develop to the blastocyst stage. Non-HIP embryos are associated with increased aneuploidy for most screened chromosomes, suggesting an association between morphology and chromosomal status. Interestingly, HIP embryos are no less likely to have abnormal numbers of chromosomes 13, 20, and 21. Our data suggest that PGD is a useful tool to detect embryos with potentially viable but detrimental chromosomal abnormalities that are not detected by embryo morphology alone.

Embryo culture to the blastocyst stage does not select for gender

Reports of increased developmental rates in male embryos have resulted in concerns that day 5 blastocyst transfer may lead to gender selection. We compared blastocyst development and prevalence of euploidy in XX vs. XY embryos. Retrospective analysis. We identified all women who underwent in vitro fertilization (IVF) with their first preimplantation genetic diagnosis (PGD) cycle for recurrent miscarriage, recurrent IVF failure, advanced maternal age, or other indications between January 1 and December 31, 2006. Fluorescence in situ hybridization was utilized to analyze 9 or 12 chromosomes. Embryos with abnormal sex chromosomes were excluded. P values were adjusted to account for lack of independence among embryos from the same woman. A total of 481 embryos from 99 cycles in 99 patients with complete results for the 9-chromosome panel were analyzed. 235 (49%) were XX and 246 (51%) were XY. The prevalence of euploidy was similar between the XX and XY embryos (30% vs. 32%, P=0.73). XX and XY embryos were equally likely to develop to the blastocyst stage by day 5 (59% vs. 60%, P=0.84), and those that became blastocysts had similar proportions of euploidy (41% vs. 37%, P=0.43). Results were similar in a subset of embryos in which all 12 chromosomes were analyzed. Abnormal numbers of individual autosomes were no more likely to occur in XX than XY embryos. XX and XY embryos are equally likely to develop to the blastocyst stage by day 5. Moreover, XX and XY blastocysts are equally likely to be euploid. There is no predilection for aneuploidy of any single autosome to occur with greater frequency in XX vs. XY embryos. Our data suggest that day 5 blastocyst culture does not result in gender selection and that euploidy and aneuploidy are not gender dependent.

Differential Delivery Rates Between Infertility Diagnosis in a PGD/FISH Program

In the majority of patients undergoing IVF, competent embryos are identified using cumulative morphology grading. However, specific populations of patients exist that fail to conceive or continue to miscarry despite multiple attempts at IVF with the same embryo grading criteria. It has been hypothesized that these patients have less chromosomally normal embryos making the option of PGD desirable to identify euploid embryos for transfer. PGD as a treatment for infertility is often indicated in patients suffering from recurrent pregnancy loss (RPL), advanced maternal age, multiple failed in-vitro fertilization (IVF) attempts, and repeat biochemical pregnancies after IVF. However, this application of PGD has demonstrated varying reports of success. The goal of this study is to analyze the relationships between embryo morphology and aneuploidy and pregnancy outcomes in each patient population. Retrospective data analysis All patients over a 15-month period who had PGD with FISH for aneuploidy were included. Patients undergoing PGD for translocation or single gene defects were excluded. The clinical pregnancy rates (CPR) and ongoing/delivery rates were recorded by age and euploidy by indication. Embryo data was recorded by pronuclear and day three morphology and percent euploid by indication. A significant difference was observed in ongoing pregnancy rates between PGD patients of specific diagnosis groups. With the exception of the advanced maternal age group, there was no difference in the percent of euploid embryos between groups. In addition, there was no significant difference between embryo morphology in any of the diagnosis groups. Tabled 1 Tabled 1 These data suggest that PGD results in high pregnancy success rates for recurrent miscarriage patients. When a previous pregnancy rate of zero percent prior to PGD is considered, PGD proves to increase pregnancy rates by a significant amount for all of the diagnosis populations. However, the large discrepancy in success rates and the lack of significantly different morphology and aneuploidy data between groups suggests the existence of other unknown factors responsible for the majority of infertility in patients with recurrent IVF failure and repeat biochemical pregnancies.

Rezidivierende Aborte und frustrane In-vitro-Fertilisation

Antibodies against annexin-V, a potent anticoagulant abundant in placental tissues, were recently controversially reported to be associated with recurrent miscarriages or failures of in-vitro-fertilization (IVF) attempts. We screened 56 women (34.7 +/- 4.3 years of age: mean +/- 1 SD) with recurrent IVF failures and/or early pregnancy losses for resistance against activated protein C, lupus anticoagulant and antibodies against annexin V, cardiolipin or beta(2)-glycoprotein-1. Among them the prevalence of APC-R (8/56, 14%) and elevated levels of IgG- or IgM-anti-cardiolipin antibodies (7/56, 12%) were more common than elevated levels of (IgG or IgM) antibodies against beta(2)-glycoprotein-1 (3/56, 5%) or annexin-V (1/56, 2%). 42 (75%) of the women had another IVF-attempt after this haemostaseological evaluation and received low molecular weight heparin and/ or acetylsalicylic acid in the case of positivity for APC-resistance, lupus anticoagulant or antibodies against annexin V, cardiolipin or beta(2)-glycoprotein-1. The outcome of these IVF-attempts were 19 pregnancies (34%): 4 early miscarriages (7%) and 15 so far uncomplicated pregnancies (27%). The only woman with an elevated anti-annexin V (IgG) level had had 7 IVF before and received 40 mg Enoxaparin (Lovenox) subcutaneously once daily during the 8(th) IVF, which resulted in a healthy pregnancy. Our findings suggest that among women with recurrent IVF failures anti-annexin V antibody positivity is less prevalent than APC-resistance, lupus anticoagulant (LA) or elevated levels of antibodies against cardiolipin, beta(2)-glycoprotein-1 and that the IVF-result of women with APC-R, LA or with elevated levels of antibodies against annexin V, cardiolipin or beta(2)-glycoprotein might be positively influenced by low molecular weight heparin.

Preimplantation genetic diagnosis by fluorescence in situ hybridization: clinical possibilities and pitfalls.

Preimplantation genetic diagnosis using the fluorescence in situ hybridization technique (FISH) is being used widely to prevent the transmission of sex-linked diseases, to screen for translocations, and for aneuploidy screening in specific in vitro fertilization (IVF) patient groups, along with FISH analysis of spermatozoa in infertile men. In this study, we aim to critically analyze our clinical results in patients at risk of transmitting sex-linked diseases (n = 55), in carriers of translocations (n = 43), in women who have recurrent miscarriage (two or more miscarriages) (n = 128), recurrent IVF failure (three or more failed IVF attempts) (n = 47), and patients of advanced maternal age (37 years old or older) (n = 79). The use of the FISH technique in carriers of sex-linked diseases and translocation patients prevents transmission of these conditions and provides good IVF outcome. In patients with recurrent miscarriage, implantation failure, and advanced maternal age, a high incidence of embryos with abnormal chromosomes 13,16,18,21,22, X, and Y was observed (range 69-75%), as expected. In those three groups of patients, the selection of euploid embryos for transfer resulted in good pregnancy rates with a low incidence of miscarriage. Limitations and pitfalls of this technique are also discussed.