Inherited defects of piRNA biogenesis cause transposon de-repression, impaired spermatogenesis, and human male infertility.

piRNAs are crucial for transposon silencing, germ cell maturation, and fertility in male mice. Here, we report on the genetic landscape of piRNA dysfunction in humans and present 39 infertile men carrying biallelic variants in 14 different piRNA pathway genes, including PIWIL1, GTSF1, GPAT2, MAEL, TDRD1, and DDX4. In some affected men, the testicular phenotypes differ from those of the respective knockout mice and range from complete germ cell loss to the production of a few morphologically abnormal sperm. A reduced number of pachytene piRNAs was detected in the testicular tissue of variant carriers, demonstrating impaired piRNA biogenesis. Furthermore, LINE1 expression in spermatogonia links impaired piRNA biogenesis to transposon de-silencing and serves to classify variants as functionally relevant. These results establish the disrupted piRNA pathway as a major cause of human spermatogenic failure and provide insights into transposon silencing in human male germ cells.

Evaluation of an Updated Gene Panel as a Diagnostic Tool for Both Male and Female Infertility.

In recent years, an increasing number of genes associated with male and female infertility have been identified. The genetics of infertility is no longer limited to the analysis of karyotypes or specific genes, and it is now possible to analyse several dozen infertility genes simultaneously. Here, we present the diagnostic activity over the past two years including 140 patients (63 women and 77 men). Targeted sequencing revealed causative variants in 17 patients, representing an overall diagnostic rate of 12.1%, with prevalence rates in females and males of 11% and 13%, respectively. The gene-disease relationship (GDR) was re-evaluated for genes due to the addition of new patients and/or variants in the actual study. Five genes changed categories: two female genes (MEIOB and TBPL2) moved from limited to moderate; two male genes (SOHLH1 and GALNTL5) moved from no evidence to strong and from limited to moderate; and SEPTIN12, which was unable to classify male infertility, was reclassified as limited. Many infertility genes have yet to be identified. With the increasing integration of genetics in reproductive medicine, the scope of intervention extends to include other family members, in addition to individual patients or couples. Genetic counselling consultations and appropriate staffing will need to be established in fertility centres. Trial registration number: Not applicable.

Ten-year outcomes of patients who developed persistent azoospermia following chemotherapy associated with different oncological diagnoses: A retrospective cohort study from a different perspective.

BACKGROUND: This study evaluated the treatment procedures for chemotherapy (CT)-induced persistent azoospermia and their outcomes from a different perspective. METHODS: In 63 patients (mean age: 30.16 ± 4.91 years) who had undergone CT 11 ± 5 years earlier, the semen volume, gonadotropins level, FSH level, genetics, micro-testicular sperm extraction (m-TESE) result, sperm DNA fragmentation index (SDFI), semen reactive oxidative stress (ROS) rate, duration of embryonic development, and pregnancy and baby take-home rates were examined. The correlations between the ROS rates and the SDFIs, m-TESE results, sperm motility, pathology scores, time-lapses, and baby take-home rates were evaluated. RESULTS: The semen volumes were 3.5 ± 1.1/ml. The FSH level following CT was 17.87 ± 5.80 mIU/ml. A sperm rate of 34.9% was found from the m-TESE result. The mean SDFI and ROS rate were 4 (<15-30>) and 1.29 ± 0.51, respectively. The time-lapse was calculated as 5h. Pregnancy and live birth were achieved at 20.63% and 12.7%, respectively. In the patients with a low ROS (≤1.42) and SDFI (≤15), the m-TESE success rate was high, the FSH value was low, the pathological score and fertilization rate were elevated, the embryonic cleavage period was normal, and the pregnancy and baby take-home rates were high. DISCUSSION: The sperms may be detected using m-TESE in patients who develop persistent azoospermia associated with CT due to different oncological diagnoses. Our study revealed that a low FSH value and normal ejaculatory ROS rates are positive predictive factors of sperm detection before m-TESE. The motility of the sperms detected after m-TESE and normal SDFI rates were found to be positive predictive criteria of high fertilization, good embryonic cleavage, pregnancy, and live birth.

Does rotation during catheter withdrawal in embryo transfer increase pregnancy rates?

BACKGROUND: The aim of this study was to determine whether there is a difference in pregnancy rates between groups that we removed directly and removed by rotating the embryo transfer catheter 360°. METHODS: The study group consisted of 552 patients who were withdrawn by 360° rotation and 797 patients who were withdrawn without catheter rotation. All patients underwent one or two fresh ETs on day 3 or 5. Groups were compared in terms of cycle characteristics and clinical pregnancy rates. RESULTS: There were no significant differences in demographic and clinical characteristics of both groups, such as age, body mass index (BMI), duration of infertility, causes of infertility, and basal hormone levels. Clinicals pregnancy rate, in the study group, 48% (265/552) and in the control group, 50.8% (405/797) were similar in both groups. When the implantation rate and miscarriage rate were examined, both groups were found to be similar. CONCLUSION: It was found that 360° rotation while pulling catheter during embryo transfer had no effect on pregnancy and clinical pregnancy.

Severe ovarian hyperstimulation syndrome and gonadotropin-releasing hormone agonist trigger in patients with hypogonadotropic hypogonadism: A report of two cases.

Ovarian Hyperstimulation syndrome (OHSS) is a rare condition in patients with hypogonadotropic hypogonadism. Two patients with hypogonadotropic hypogonadism are reported, a rare case of severe OHSS and a case of prevented OHSS via gonadotropin-releasing hormone (GnRH) agonist trigger, respectively. The first case was a 31-year-old patient. In vitro fertilization (IVF) treatment was performed three times but the patient never developed OHSS. The first patient was diagnosed as having severe OHSS on the ninth day after the fresh embryo transfer. She stayed 66 days in hospital and 50.5 litres of fluid were aspirated from her abdomen. The second case was a 26-year-old and primary infertile patient. She had never undergone IVF treatment. The GnRH agonist stimulation test was performed before IVF treatment. After the ovarian stimulation, GnRH agonist trigger was given. Thirty-two oocytes were retrieved from the ovaries and OHSS did not occur. Although severe OHSS is rare, it can develop in patients hypogonadotropic hypogonadism. If a GnRH stimulation test is performed before ovarian stimulation, OHSS can be prevented because the test allows agonist triggering instead of hCG in hypogonadotropic hypogonadism.

The Effects of Microfluidic Sperm Sorting, Density Gradient and Swim-up Methods on Semen Oxidation Reduction Potential.

PURPOSE: To compare the effects of microfluidic sperm sorting, density gradient and swim-up methods on the oxidative reduction potential (ORP) of split semen samples from a single patient population. MATERIALS AND METHODS: A prospective controlled study was conducted to compare the effects of three different semen processing methods using split semen samples from the same population of infertile men. The primary outcome was the ORP. Secondary outcomes were the sperm concentration, progressive motility rate and total sperm motility. RESULTS: A total of 57 split semen samples were included in this study. The ORP was significantly lower in the microfluidic group compared to the density gradient and swim-up groups (p < 0.05). The ORP/sperm concentration ratio was significantly lower in the microfluidic and density gradient groups compared to the swim-up group (p < 0.05).Total sperm concentration was significantly higher in the density gradient group than the microfluidic and swim-up groups (p < 0.05). Motility was significantly higher in the microfluidic and swim-up groups than the density gradient group (p < 0.05). The progressive motile sperm rate was significantly higher in the microfluidic and swim-up groups than the density gradient group (p < 0.05). CONCLUSIONS: Microfluidic sperm sorting was better for selecting highly motile sperm and yielded a lower ORP than conventional sperm preparation methods.

Non-Invasive Isthmocele Treatment: A New Therapeutic Option During Assisted Reproductive Technology Cycles?

BACKGROUND: The objective of the study was to evaluate a new medical treatment strategy for infertile patients with isthmocele. METHODS: This was a retrospective evaluation of the records of infertile patients with symptomatic isthmocele who received non-invasive isthmocele treatment (NIIT) before in vitro fertilization (IVF) treatment cycles. Isthmocele volumes were measured before and after NIIT. The IVF results and isthmocele-related complaints were also analyzed. The patients were treated with a depot gonadotropin-releasing hormone agonist for 3 months before frozen-thawed embryo transfer cycles. RESULTS: The mean isthmocele volume was 471.06 ± 182.81 mm3 (range: 289.43 - 765.4 mm3) in fresh cycles, but was reduced to 47.94 ± 29.48 mm3 (range: 18.70 - 105.6 mm3) in frozen-thawed cycles (P < 0.05). Intrauterine fluid was observed in two patients during fresh cycles, but was absent after NIIT during frozen-thawed cycles. There was no brown bloody discharge on the tip of the embryo transfer catheter in any case after NIIT. Two patients became pregnant and underwent term cesarean delivery (25%). CONCLUSIONS: NIIT can serve as an alternative pretreatment option for patients with isthmocele during IVF cycles.

Dydrogesterone-primed ovarian stimulation is an effective alternative to gonadotropin-releasing hormone antagonist protocol for freeze-all cycles in polycystic ovary syndrome.

OBJECTIVE: To compare the effects of progestin-primed ovarian stimulation using dydrogesterone (DYD) and a gonadotropin-releasing hormone (GnRH) antagonist protocol on cycle characteristics and pregnancy rates in freeze-all cycles in patients with polycystic ovary syndrome (PCOS). METHODS: Medical records of PCOS patients who underwent freeze-all in vitro fertilization cycles between April 2017 and April 2019 at the Novafertil in vitro fertilization Center were retrospectively evaluated. The primary outcome measure was the incidence of premature luteinizing hormone surge. Secondary outcome measures were the total number of mature oocytes retrieved, fertilization rate, clinical pregnancy rates and ongoing pregnancy rates. RESULTS: A total of 525 patients were included in the study. DYD-primed ovarian stimulation and a GnRH antagonist protocol were applied in 258 and 267 patients, respectively. The baseline parameters were similar between the two groups. The numbers of mature and fertilized oocytes were similar in the cetrorelix (CET) group and DYD group (11.43 ± 3.48 vs. 11.29 ± 4.34, respectively, P = 0.692; and 8.98 ± 2.93 vs. 8.62 ± 3.67, respectively, P = 0.208). Premature luteinization was rare in both groups, and the difference between the groups was not statistically significant (2.9% vs. 1.5%, respectively, P = 0.268). There was no significant difference in clinical pregnancy rate of the first frozen embryo transfer cycle between the DYG group and the CET group (56% [120/214] vs. 55.6% [113/203], respectively, P = 0.283). There were no significant differences in biochemical pregnancy rates, implantation rates, miscarriage rates or ongoing pregnancy rates between the two groups (P > 0.05). CONCLUSION: Dydrogesterone-primed ovarian stimulation seems to be an effective alternative to the GnRH antagonist protocol for freeze-all cycles in PCOS patients.

Evaluation from a different perspective of 10-year results of infertile males with Y chromosome AZFc microdeletions compared with a control group.

AZFc microdeletions will be evaluated upon being divided into partial and complete subgroups. The association of deletions with reactive oxidative stress (ROS) and sperm DNA fragmentation (SDFI) and the impact of their coexistence on fertility starting from the pregnancy process until live birth will be presented. Semen analyses, microbiological results, hormones, ROS and sperm TUNEL tests were checked. Preimplantation genetic testing (PGT) was planned for relevant patients. Intracytoplasmic sperm injection (ICSI) was applied. Their embryo fragmentation was monitored via time lapse. Their results were compared with those with no AZF deletion and no other genetic problems. Azoospermia rate was 71.5%, m-TESE success rate was 25%, pregnancy rate was 26% and live child rate was 2.2%. No difference was detected between the partial and total groups in terms of ROS and SDFI rates and no difference was identified with the control group. Better results were obtained in terms of live child rate in patients with partial AZFc and low ROS/SDFI. Spermatozoon was retrieved in AZFc deletions and pregnancy, and live child was identified. No AZFc impact was observed on ROS and SDFI in the results compared with the control groups in terms of their coexistence.

Comparison of microfluid sperm sorting chip and density gradient methods for use in intrauterine insemination cycles.

OBJECTIVE: To compare the effect of microfluiding sperm sorting chip and density gradient methods on ongoing pregnancy rates (PRs) of patients undergoing IUI. DESIGN: Retrospective cohort study. SETTING: Hospital IVF unit. PATIENT(S): Couples with infertility undergoing IUI cycles between 2017 and 2018. INTERVENTION(S): Not applicable. MAIN OUTCOME MEASURE(S): Ongoing PRs. RESULT(S): A total of 265 patients were included in the study. Microfluid sperm sorting and density gradient were used to prepare sperm in 133 and 132 patients, respectively. Baseline spermiogram parameters, including volume, concentration, motility, and morphology, were similar between the two groups. Total motile sperm count was lower in the microfluiding sperm sorting group at baseline (35.96 ± 37.69 vs. 70.66 ± 61.65). After sperm preparation sperm motility was higher in the microfluid group (96.34 ± 7.29 vs. 84.42 ± 10.87). Pregnancy rates were 18.04% in the microfluid group and 15.15% in the density gradient group, and ongoing PRs were 15.03% and 9.09%, respectively. After using multivariable logistic regression and controling for confounding factors, there was a significant increase in ongoing PRs in the microfluid sperm sorting group. The adjusted odds ratio for ongoing pregnancy in the microfluid group compared with the density gradient group was 3.49 (95% confidence interval 1.12-10.89). CONCLUSION(S): The microfluid sperm sorting method significantly increased the ongoing PRs compared with the density gradient group in IUI cycles.

Iatrogenic ureteral obstruction during transvaginal ocyte retrieval

ABSTRACT Transvaginal oocyte retrieval is a crucial step in assisted reproductive technology. Various complications may arise during this procedure. Ureteral injury is a rare, but a serious complication in gynecological practice. During oocyte retrieval, ureteral injuries, detachment and obstruction can be seen, though rare. In this study, we will present ureteral obstruction that develops secondary to small hematoma, which mimics ovarian cyst torsion or ruptured ovarian cyst.

Iatrogenic Ureteral Obstruction During Transvaginal Oocyte Retrieval.

Transvaginal oocyte retrieval is a crucial step in assisted reproductive technology. Various complications may arise during this procedure. Ureteral injury is a rare, but a serious complication in gynecological practice. During oocyte retrieval, ureteral injuries, detachment and obstruction can be seen, though rare. In this study, we will present ureteral obstruction that develops secondary to small hematoma, which mimics ovarian cyst torsion or ruptured ovarian cyst.

Rare genetic variants potentially involved in ovarian hyperstimulation syndrome.

PURPOSE: We aim to investigate whether there is a genetic predisposition in women who developed ovarian hyperstimulation syndrome (OHSS) after GnRH antagonist protocol with GnRH agonist trigger and freeze-all approach. METHODS: Four patients with OHSS after GnRH agonist trigger and freeze-all approach were gathered from the worldwide patient population. These patients were analyzed through Whole Exome Sequencing. In this study known causes of OHSS were investigated and new causes present in at least two individuals were searched for. RESULTS: In the first part of the study, we evaluated the presence of mutations in genes already known to be involved in OHSS. In PGR and TP53, heterozygous alterations were detected. PGR is predicted to be involved in progesterone resistance with a recessive inheritance pattern and is, therefore, not considered as being causal. The consequences of the variant detected in TP53 currently remain unknown. In part 2 of the study, we assessed the clinical significance of variants in genes previously not linked to OHSS. We especially focused on genes with variants present in ≥ 2 patients. Two patients have variants in the FLT4 gene. Mutations in this gene are linked to hereditary lymphedema, but no link to OHSS has been described. CONCLUSIONS: Defining a genetic predisposition for OHSS is essential in view of prevention. In this study, a potential link between the FLT4 gene and OHSS has been suggested. Future functional studies are essential to define a more precise involvement of the detected variants in the development of OHSS.

Dipeptidyl peptidase-4 and adenosine deaminase enzyme levels in polycystic ovary syndrome.

Polycystic ovary syndrome (PCOS) is one of the most common endocrinopathies in reproductive age women and insulin resistance (IR) and hyperinsulinism play a critical role in the pathogenesis. Glucagon-like peptide-1 (GLP-1), promotes insulin secretion, inhibits glucagon secretion. GLP-1 is degraded by dipeptidyl peptidase-4 (DPP-4). DPP-4, also interacts with adenosine deaminase (ADA). Therefore, IR may have a significant connection with ADA activity. The aim of this study is to compare levels of DPP-4 and ADA enzymes in PCOS and infertile patients. Forty-four patients with PCOS and 44 infertile patients with normal ovarian reserve were enrolled in the study. Serum ADA, DPP-4, AMH, glucose and insulin levels were measured. HOMA-IR method was used to assess insulin sensitivity. ADA, DPP-4, AMH, HOMA-IR (p < .05) and insulin levels (p < .01) were found to be increased in PCOS patients. Considering all study participants AMH levels were found to be positively correlated with ADA (r: 0.734) and DPP-4 (r: 0.449) levels. Also ADA levels were found to be positively correlated with DPP-4 (r: 0.472), insulin (r: 0.216) and HOMA-IR (r: 0.223). Our findings about the elevation of DPP-4 levels in patients with PCOS suggest that the use of DPP-4 inhibitors may be beneficial in treatment of these patients.

Comparison of embryo morphokinetics following intracytoplasmic sperm injection in smoker and non-smoker couples: Are the results different?

OBJECTIVE: To assess early embryo development via time-lapse in smokers and non-smokers. METHODS: The retrospective study was conducted at Novafertil IVF centers in Konya, Turkey and comprised oocytes of both smoker and non-smoker couples subjected to in vitro fertilisation / introcytoplasmic sperm injection from 2012 to 2015. Age, basal follicle-stimulating hormone, number of stimulation days, amount of gonadotropin used, number of metaphase II oocytes, number of embryos transferred and pregnancy, abortus and clinical pregnancy rates were noted. The embryos were observed for 72 hours in the time-lapse monitoring system. SPSS 22 was used for data analysis. RESULTS: Of the 257 couples, 132(51.4%) were non-smokers and 125(48.6%) were smokers. A total of 1,414 oocytes were collected from non-smokers and 1,280 oocytes from smokers. There was no significant difference in the age of patients and number of stimulation days between the smoker and non-smoker groups (p>0.05). The number of oocytes, fertilised oocytes, transferred embryos and metaphase II oocytes was significantly less in the smoker group (p<0.05). The rate of pregnancy and ongoing pregnancy was also lower in the smoker group (p<0.05). A difference was observed in time of pronuclei appearance, t8, t9+ cleavage times in time-lapse in the smoker and non-smoker groups (p<0.05). A prolongation was observed in time to pronuclear fading and t2 cleavage times in time-lapse in the non-smoker group (p<0.05). Some chromosomal number and structural defects were identified in pre-implantation genetic in some embryos with prolongation in time-lapse cleavage time in the smoker and non-smoker groups. CONCLUSIONS: The negative impacts of smoking were not observed at each cleavage phase of embryo development in time-lapse.

Is There Any Impact of Copper Intrauterine Device on Female Sexual Functioning?

INTRODUCTION: Intrauterine Device (IUD) is the most preferred modern contraceptive method in Turkey. Female Sexual Dysfunction (FSD) is defined as lack of one or more of the components in the sexual response cycle which includes sexual desire, impaired arousal and inability achieving an orgasm or pain with intercourse. FSD has multi-factorial aetiology. Advanced age and menopause, fatigue and stress, psychiatric and neurologic disease, childbirth, pelvic floor or bladder dysfunction, endometriosis, uterine fibroids, hypertension obesity, medication and substances, hormonal contraceptives, relationship factors are known risk factors for FSD. AIM: To investigate if IUD has any impact on female sexual functioning. MATERIALS AND METHODS: In this cross-sectional study subjects were divided into two groups. Study group consisted of 92 IUD-users (mean 5.1±1.2 years) and the control group consisted of 83 women with no contraception. Female Sexual Function Index (FSFI) questionnaire was performed to both two groups. Women with a total score lower than 26.5 were considered as having sexual dysfunction. RESULTS: The prevalence of FSD was 57.1% among participants. IUD users had a lower total FSFI score comparing to control group but the difference was not statistically different (p=0.983). A positive correlation was found between total FSFI score and duration of IUD (p=0.003). CONCLUSION: No difference was found in terms of sexual dysfunction between IUD users and women with no contraception. The prevalence of FSD was very high in both groups which may be attributed to the socio-cultural factors such as embarrassment of women due to conservatism.

GnRH agonist triggering affects the kinetics of embryo development: a comparative study.

BACKGROUND: To evaluate the effects of an ovulation triggering agent, human chorionic gonadotropin (hCG), versus a gonadotropin-releasing hormone agonist (GnRHa) on early embryo development in vitro using a time-lapse system. METHODS: Retrospective analysis of a prospectively collected database. A total of 739 embryos from 152 infertile couples undergoing intracytoplasmic sperm injection cycles. INTERVENTIONS: Embryo culture in a time-lapse incubator (EmbryoScope, Vitrolife, Göteborg, Sweden). MAIN OUTCOME MEASURES: Embryo morphokinetic parameters. RESULTS: In the 152 women, 252 embryos were derived from GnRHa-triggered cycles compared with 487 embryos derived from hCG-triggered cycles. Time-lapse analysis revealed that embryos from cycles triggered by a GnRHa cleaved faster than embryos derived from hCG-triggered cycles. CONCLUSION: Triggering with a GnRHa in in vitro fertilization cycles affects embryo kinetics.

Is It Possible to Prevent Ovarian Hyperstimulation Syndrome by Gonadotropin-Releasing Hormone Agonist Triggering and Modified Luteal Support in Patients With Polycystic Ovarian Morphology?

BACKGROUND: Gonadotropin-releasing hormone (GnRH) agonist triggering plus 1,500 IU human chorionic gonadotropin (hCG) supplementation protocol was previously claimed effective in reducing the ovarian hyperstimulation syndrome (OHSS) incidence in high responders. METHODS: This retrospective study included women with polycystic ovarian (PCO) morphology who were at high risk of OHSS and were given the GnRH agonist trigger plus hCG luteal support protocol in a single center. RESULTS: The mean peak estradiol level was 5,336 ± 2,341 (1,187 - 19,746) pg/mL. The mean number of follicles > 12 mm on the day of trigger was 22 ± 7 (9 - 51). A total of 88 cycles were undertaken. Sixty-three (71.5%) women underwent fresh embryo transfer. Fresh embryo transfer was canceled in 21 (23.8%) and embryo transfer was canceled in four (4.5%) women. The overall clinical pregnancy rate was 46.4% per started cycle. A total of 12 (13.6%) patients developed OHSS. "Freeze-all" policy did not attenuate OHSS in four patients, and three of these patients developed OHSS despite 1,500 IU hCG was not administered. CONCLUSION: We conclude that OHSS may still occur with the use of a GnRH agonist trigger combined with low-dose hCG supplementation protocol in women with polycystic ovary syndrome (PCOS) or PCO morphology. Furthermore, we also conclude that "freeze-all" policy also will not completely eliminate OHSS development in high-risk women.

Absence of luteal phase defect and spontaneous pregnancy in IVF patients despite GnRH-agonist trigger and "freeze all policy" without luteal phase support: a report of four cases.

Human chorionic gonadotropin (hCG) is commonly used for final oocyte maturation in "in vitro fertilization" (IVF)-treatment cycles, however, the main important risk is development of severe ovarian hyperstimulation syndrome (OHSS). OHSS can almost be avoided by using gonadotrophin-releasing-hormone agonist for final oocyte maturation in an antagonist protocol. However, primarily this approach lead to a very poor reproductive outcome, despite the use of a standard luteal phase support. The reason seems to be severe luteolysis. Obviously, luteolysis post-gonadotropin-releasing-hormone-agonist (post-GnRH-a) trigger is individual specific, and not all patients will develop a complete luteolysis, as expected previously. Luteolysis can been reverted by the administration of hCG. Unprotected intercourse around the time of ovulation induction and oocyte retrieval can lead to a spontaneous conception in IVF treatment and, endogenous hCG, produced by the trophoblast, will rescue the corpora lutea. Therefore, one should not rely on complete luteolysis after GnRH-a triggering and, especially patients for egg donation and pre-implantation-genetic diagnosis for single gene disorder, have to be counselled to avoid unprotected intercourse.

Early Morphokinetic Monitoring of Embryos after Intracytoplasmic Sperm Injection with Fresh Ejaculate Sperm in Nonmosaic Klinefelter Syndrome: A Different Presentation.

The patient was diagnosed with nonmosaic 47, XXY Klinefelter Syndrome with the AZF deletion absent and SRY+. The nonmosaic 47, XXY karyotype was confirmed on a skin biopsy chromosomal analysis. Using only ejaculate motile sperms, 11 oocytes underwent ICSI and were placed rapidly in a time lapse (Embryoscope ©) with a specific culture dish. Biopsies were performed on six embryos on the 3rd day, and numerical chromosomal abnormalities were observed using the FISH test before transfer. PGS results were normal in only two embryos with normal morphokinetics in the Embryoscope. For clinical confirmation of pregnancy, ultrasonographic examination was performed during the 7th week of pregnancy, and two gestational sacs and fetal heart beat were observed.