Intracytoplasmic Sperm Injection Failure Research Articles

Impact of granulocyte colony stimulating factor infusion on the implantation rate in women with unexplained previous intracytoplasmic sperm injection failure: a case control study

Impact of granulocyte colony stimulating factor infusion on the implantation rate in women with unexplained previous intracytoplasmic sperm injection failure: a case control study

Elucidating the impact of Y chromosome microdeletions and altered gene expression on male fertility in assisted reproduction.

Genetic abnormalities like Y chromosome microdeletions are implicated in male infertility. This study investigated the association of azoospermia factor (AZF) region microdeletions with unsuccessful assisted reproductive techniques (ART), including in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). This cross-sectional analysis study examined 80 Iranian oligospermic men (mean age 34years) with prior failed ICSI and IVF cycles (IR.IAU.TNB.REC.1401.041). Semen analysis evaluated quantity/quality parameters based on World Health Organization guidelines. Participants were stratified by sperm DNA fragmentation (SDF) levels into: control (SDF < 15%, n = 20), mild elevation (15% ≤ SDF ≤ 30%, n = 60), and high (SDF > 30%, n = 20). Multiplex PCR mapped AZF microdeletions in the high SDF group. The AZF-associated genes were selected by RNA Seq analysis, and the candidate genes were checked for expression level by real-time PCR. High SDF individuals exhibited poorer semen metrics, including 69% lower sperm concentration (P = 0.04) than those without SDF. Of this subset, 45% (9/20 men) harboured predominately AZF microdeletions. Men with AZF microdeletions showed higher SDF (32% vs 21%, P = 0.02) and altered AZF-associated genes expression. As USP9Y 3-fold, UTY 1.3-fold, and BPY2 1-fold revealed up-regulation, while IQCF1 8-fold, CDY 6.5-fold, DAZ 6-fold, and DDX3Y 1-fold underwent down-regulation. The PAWP gene was also down-regulated (5.7-fold, P = 0.029) in the IVF/ICSI failure group. AZF microdeletions significantly impact male infertility and ART outcomes. High SDF individuals exhibited poorer semen metrics, with 45% AZF microdeletions. These microdeletions altered AZF-associated genes expression, affecting fertility mediator PAWP independently. Dual AZF and SDF screening enables personalized management in severe male infertility, potentially explaining IVF/ICSI failures.

Stress Biomarkers Transferred Into the Female Reproductive Tract by Seminal Plasma Are Associated with ICSI Outcomes.

This study aimed to determine whether male stress is related to seminal stress biomarkers and pregnancy achievement in women exposed to their partner's seminal plasma (SP) in the intracytoplasmic sperm injection (ICSI) cycle. In this pilot prospective study, 20 couples undergoing ICSI, as well as 5 fertile sperm donors and 10 saliva donors, were investigated. Women were exposed to their partner's SP via unprotected sexual intercourse during the ICSI cycle and intravaginal application on the day of ovum pick-up (Day-OPU). Semen samples were collected from male partners by masturbation on the Day-OPU. Saliva and serum samples were collected prior to masturbation. Body fluids were frozen at - 80°C until assayed. Biomarkers of activity of the sympathetic adrenomedullary axis (salivary alpha-amylase and adrenaline), sympathetic neural axis (noradrenaline and dopamine), hypothalamic-pituitary-adrenal (HPA) system (cortisol), and immune system (C-reactive protein and interleukin (IL)-18) were estimated to examine their association with SP composition and clinical pregnancy achievement. The clinical pregnancy rate was 45.0%. In the unsuccessful ICSI group, blunted levels of salivary and serum cortisol were found compared to the successful ICSI group and the fertile sperm donors. With regard to seminal markers, decreased cortisol level and elevated noradrenaline, noradrenaline/cortisol ratio, and lL-18 levels were strongly associated with ICSI failure (areas under the ROC curves were, 0.813, 0.848, 0.899, and 0.828, respectively). These findings confirm that stress response systems activity affects SP composition, which in turn is associated with ICSI outcomes in women exposed to their partner's SP during an ICSI cycle.

ICSI using testicular spermatozoa after failure of ICSI with ejaculated spermatozoa could be a good choice: A propensity score‐matched cohort study

AbstractBackgroundEjaculated spermatozoa are considered to possess a higher fertilisation potential than testicular spermatozoa. In selected cases, the use of testicular spermatozoa from non‐azoospermic infertile men resulted in a higher implantation and pregnancy rate than the use of ejaculated spermatozoa.ObjectiveThe primary objective was to compare the live birth rate and cumulative live birth rate between couples with failed intracytoplasmic sperm injection procedure using ejaculated spermatozoa who subsequently had an intracytoplasmic sperm injection cycle with testicular spermatozoa and those who subsequently had an intracytoplasmic sperm injection cycle with ejaculated spermatozoa. The secondary objective was to determine the indications for the use of testicular spermatozoa after intracytoplasmic sperm injection failure with ejaculated spermatozoa.Materials and methodsA retrospective study of matched couples using propensity score matching analysis was performed. After an intracytoplasmic sperm injection failure (cycle_1), intracytoplasmic sperm injection with either ejaculated spermatozoa (ejaculated sperm group), or testicular spermatozoa (testicular sperm group), was performed (cycle_2). The matching was on intracytoplasmic sperm injection performed in cycle_1 according to spermatozoa used (testicular or ejaculated) in cycle_2. Logistic regression was used to evaluate the influence of sperm origin on cumulative live birth rate. Univariate analysis on parameters of cycle_1 was used to identify the prognostic factors to propose an intracytoplasmic sperm injection with testicular spermatozoa in case of cycle_1 failure. The study outcomes were live birth rate and cumulative live birth rate.ResultsAmong the 6034 couples available, 63 were selected to constitute the testicular sperm group and 63 were selected by propensity score matching to constitute the ejaculated sperm group. After matching, the DNA fragmentation index was higher in the testicular sperm group (13.43% ± 9.65% vs. 8.93% ± 4.47%, p = 0.013); no significant difference was observed for the fertilisation rate, the number of obtained embryos, blastulation rate and frozen embryo rate. In cycle_2, the live birth rate was higher in the testicular group (22.2% vs. 0.0%, p &lt; 0.001), as was the cumulative live birth rate (25.4% vs. 6.3%, p = 0.065). The prognostic factors identified for the proposal of intracytoplasmic sperm injection procedure with testicular spermatozoa after intracytoplasmic sperm injection failure with ejaculated spermatozoa were: teratozoospermia, cryptozoospermia and high DNA fragmentation index.DiscussionAccording to the present study and current knowledge, the use of testicular spermatozoa after failed intracytoplasmic sperm injection procedure in non‐azoospermic men could be proposed instead of sperm donation in case of high sperm DNA fragmentation index, cryptozoospermia and teratozoospermia. A good oocyte response to ovarian stimulation during the previous assisted reproductive technology attempt will increase the chance of success. Although the main limitation of the current study is its retrospective nature, the use of the propensity score matching to perform causal inference study increases its reliability.ConclusionThe present study supports that the use of testicular spermatozoa outside the classical indication of azoospermia is a good option when the indication is well established. However, before proposing a testicular biopsy, an improvement in sperm characteristics should be considered by treating the causes of sperm alteration.

Defects in phospholipase C zeta cause polyspermy and low fertilization after conventional IVF: not just ICSI failure.

Phospholipase C zeta (PLCζ) is a key sperm-borne oocyte-activating factor that triggers Ca2+ oscillations and the subsequent block to polyspermy following gamete fusion. Mutations in PLCZ1, the gene encoding PLCζ, cause male infertility and intracytoplasmic sperm injection (ICSI) fertilization failure; and PLCζ expression and localization patterns are significantly correlated with ICSI fertilization rate (FR). However, in conventional in vitro fertilization (cIVF), whether and how sperm PLCζ affects fertilization remain unclear. Herein, we identified one previously reported and two novel PLCZ1 mutations associated with polyspermy in vitro that are characterized by excessive sperm-zona binding and a delay in pronuclei (PN) formation. Immunofluorescence staining and oocyte activation testing revealed that virtually all spermatozoa from patients lacked functional PLCζ and were thus unable to evoke Ca2+ oscillations. ICSI with an artificial oocyte activation treatment successfully rescued the polyspermic phenotype and resulted in a live birth. Furthermore, we analyzed PLCζ in an additional 58 males after cIVF treatment in the Reproductive and Genetic Hospital of CITIC-Xiangya (Changsha, China) between February 2019 and January 2022. We found that the proportion of spermatozoa that expressed PLCζ was positively correlated with both 2PN rate and total FR. The optimal cutoff value below which males were likely to experience low FR (total FR ≤30%) after cIVF was 56.7% for the proportion of spermatozoa expressing PLCζ. Our study expands the mutation and the phenotypic spectrum of PLCZ1 and further suggests that PLCζ constitutes a promising biomarker for identifying low FRs cases in cIVF due to sperm-related oocyte activation deficiency and that sperm PLCζ analysis may benefit the wider male population and not only men with ICSI failure.

The role of testicular sperm extraction in repeated intracytoplasmic sperm injection failures using ejaculate sperm

The role of testicular sperm extraction in repeated intracytoplasmic sperm injection failures using ejaculate sperm

P-058 Defects in phospholipase C zeta cause polyspermy and low fertilization after conventional IVF: not just ICSI failure

Abstract Study question How does sperm phospholipase C zeta (PLCζ) affect fertilization in conventional in vitro fertilization (cIVF)? Summary answer Absence of PLCζ causes polyspermy, and a decline in the proportion of sperm expressing PLCζ is correlated with a low fertilization rate (FR) after cIVF. What is known already PLCζ is a key sperm-borne factor that triggers Ca2+ oscillations and the subsequent oocyte activation following gamete fusion. Mutations in PLCZ1, the gene encoding PLCζ, cause male infertility and intra-cytoplasmic sperm injection (ICSI) fertilization failure; and PLCζ expression and localization patterns are significantly correlated with ICSI FR. However, few studies have been published on the relationship between PLCζ and cIVF, an insemination procedure involving several key events that are bypassed in ICSI, e.g., sperm-zona binding and penetration, as well as polyspermy blocking. It is possible that sperm PLCζ may affect fertilization in a different manner from that in ICSI. Study design, size, duration Sixty couples who underwent cIVF treatments were recruited from the Reproductive and Genetic Hospital of CITIC-Xiangya between February 2019 and January 2022. Participants/materials, setting, methods We performed whole-exome sequencing in two unrelated males who exhibited infertility and polyspermy in cIVF. Immunofluorescence staining and oocyte-activation testing were employed to evaluate the effects of the candidate variants on PLCζ protein. Subsequent PLCζ analysis was performed in additional 58 males who underwent cIVF. The Pearson correlation coefficient was performed to evaluate the relationships between PLCζ and FRs. The receiver operating characteristic curve and Youden’s index analysis were used to indicate the cutoff value. Main results and the role of chance We identified one previously reported and two novel PLCZ1 variants in two males that were associated with male infertility and polyspermy characterized by excessive sperm–zona binding and a delay in pronuclear (PN) formation. In vitro functional analyses revealed that virtually all sperm from patients lacked functional PLCζ and were thus unable to evoke the physiologic pattern of Ca2+ oscillations. ICSI with an artificial oocyte-activation treatment successfully rescued the polyspermic phenotype and resulted in a live birth. Subsequent PLCζ analysis were performed in additional 58 males, who were allocated to three groups according to fertilization outcomes: a multiple pronuclear (MPN) group (no. of PN ≥ 3, MPN rate ≥ 50%, n = 9) and low fertilization (LF, total FR ≤ 30%, n = 18) and normal fertilization groups (NF, 2PN ≥ 50%, 1PN + MPN &amp;lt; 30%, n = 31). We found that the proportion of sperm that expressed PLCζ was positively correlated with both 2PN rate (P = 0.0005) and total FR (P = 0.0008) after cIVF, and that the optimal cutoff value below which males were likely to experience low FR (total FR ≤ 30%) after cIVF was 56.7% (area under the ROC curve, 0.705; P = 0.0176). Limitations, reasons for caution Precise mechanisms that underly the roles of PLCζ in cIVF outcomes are still uncovered. Follow-up studies with larger numbers of patients are required to validate the sensitivity, specificity, and accuracy of PLCζ as a diagnostic biomarker in cIVF outcomes. Wider implications of the findings Our study identified PLCZ1 as a novel causative gene of polyspermy in humans,highlighting the potential value of PLCζ as a fertility marker for cIVF outcomes. Trial registration number nor applicable

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.&#x0D; 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.&#x0D;

Outcome analysis of ICSI assisted pregnancy using testicular sperm versus ejaculated sperm in man with severe oligozoospermia in the same ART cycle: A case report.

Intracytoplasmic sperm injection (ICSI) has become the most common method for couples with male factor infertility, and source of sperm for the procedure have evolved over time. but few have examined testicular sperm extraction vs. ejaculated sperm use for severe oligozoospermia in the same assisted reproductive technology (ART) cycle. Here, we evaluated the clinical outcomes after ICSI with testicular sperm or ejaculated in man with severe oligozoospermia in the same ART cycle. A couple who had failed the first ART cycle with ejaculated sperm, using the freshly ejaculated sperm and testicular sperm for ICSI during the second ART cycle by lack of enough sperm to fertilize in an ICSI attempt. The patient was diagnosed with severe oligozoospermia, and routine semen analysis revealed sperm concentration is less than 2 million/mL. The patient using testicular sperm versus ejaculated sperm with ICSI assisted pregnancy in the same ART cycle. We found that superior cleavage rate, number of embryos transferred and blastocyst rate with the use of testicular rather than ejaculated sperm-ICSI in the couple. The results described here suggest that use of testicular sperm may improve biologic outcomes, especially for couples with male-partner oligozoospermia who previous ICSI failures. Our case report supported the efficacy of testicular sperm preference over ejaculated sperm for ICSI in men with severe male factor infertility. It is a paradigm shift concerning the use of ejaculated sperm as the preferable source of sperm for ICSI, add to the small amount of literature on testicular sperm extraction vs. ejaculated sperm use for severe oligozoospermia in the same ART cycle.

Association of paternal MTHFR polymorphisms (C677T) with clinical outcomes in ICSI treatment.

This study aims to investigate the association between paternal methylenetetrahydrofolate reductase (MTHFR) polymorphisms (C677T) and embryonic development, pregnancy, and neonatal outcomes in intracytoplasmic sperm injection (ICSI) treatment. A total of 191 infertile men undergoing ICSI treatment at the Reproductive and Genetic Hospital, The First Affiliated Hospital of USTC, were recruited between January 2020 and June 2021. The MTHFR C677T polymorphism genotyping was evaluated in these male patients, and they were stratified into three groups according to genotyping results: Control (CC), heterozygote mutated (CT), and mutated homozygote (TT). In addition, we conducted a comparative analysis of embryonic development, pregnancy, and neonatal outcomes among these three groups. The embryonic development (including normal fertilization rate (80.14% vs. 83.06% vs. 85.10%; p = 0.37), high-quality embryo rate (45.26% vs. 43.69% vs. 46.04%; p = 0.72), blastocyst formation rate (42.47% vs. 43.18% vs. 39.38%; p = 0.62), implantation rate (42.47% vs. 36.25% vs. 41.22%; p = 0.62), and clinical pregnancy rate (64.71% vs. 58.75% vs. 66.67%; p = 0.59) were not comparable among these three groups. Moreover, no significant difference was observed in terms of pregnancy outcomes (including miscarriage rate (24.24% vs. 12.77% vs. 22.5%; p = 0.35) and live birth rate (49.02% vs. 51.25% vs. 51.66%; p = 0.96)). Additionally, no marked difference was observed in terms of neonatal outcome (including, preterm delivery rate (24% vs. 14.63% vs. 9.67%; p = 0.35), birth height (p = 0.75), birth weight (p = 0.35), neonatal sex (p = 0.48), gestational age at delivery (p = 0.24), Apgar score (p = 0.34), and birth defects (0% vs. 2% vs. 9%; p = 0.23) among the study groups. The paternal MTHFR C677T polymorphism is not associated with embryo quality, pregnancy, or neonatal outcomes in ICSI treatment. Therefore, in our population, MTHFR polymorphisms do not provide helpful information in explaining ICSI failure.

Switching to testicular sperm after a previous ICSI failure with ejaculated sperm significantly improves blastocyst quality without increasing aneuploidy risk.

The use of testicular sperm is confined to patients with azoospermia, but there is evidence to support its use in males with poor semen parameters and/or previous intracytoplasmic sperm injection (ICSI) failures with ejaculated spermatozoa. We compared the aneuploidy rate and quality between embryos derived from ICSI cycles with ejaculated sperm (EJ-ICSI) and those from ICSI cycles using testicular spermatozoa (TT-ICSI) within the same couple. Retrospective study of 27 couples who first underwent an EJ-ICSI cycle that did not result in a livebirth and afterwards a TT-ICSI cycle. Only the two closer cycles of each couple were included. Preimplantation genetic test for aneuploidies (PGT-A) was performed in both ICSI cycles and classic parameters of embryo quality were assessed until blastocyst-stage. A total of 375 embryos from 54 ICSI cycles were evaluated. Aneuploidy rate was measured by two different parameters. Patients undergoing TT-ICSI presented a similar aneuploidy rate as EJ-ICSI group: 30.7% (23.4-38.0) vs 26.8% (18.1-35.5) per inseminated oocytes (P>0.05), and 76.2% (66.2-86.2) vs 72.1% (59.1-85.2) per the total number of biopsied embryos (P>0.05), respectively. Further, the good-quality blastocyst rate per correctly fertilized oocyte was significantly higher in TT-ICSI group (33.6% (30.4-36.9)) than EJ-ICSI group (24.2% (20.3-28.0)) (P<0.001). Switching to testicular sperm for ICSI yielded better-quality blastocysts without affecting the chromosomal load of the embryos in non-azoospermic couples with a previous unsuccessful ICSI using ejaculated sperm. This strategy is a good option for couples seeking a livebirth who do not want to use donor sperm.

A novel heterozygous variant in PANX1 is associated with oocyte death and female infertility.

Oocyte death is a severe clinical phenotype that causes female infertility and recurrent in vitro fertilization and intracytoplasmic sperm injection failure. We aimed to identify pathogenic variants in a female infertility patient with oocyte death phenotype. Sanger sequencing was performed to screen PANX1 variants in the affected patient. Western blot analysis was used to check the effect of the variant on PANX1 glycosylation pattern in vitro. We identified a novel PANX1 variant (NM_015368.4 c.86G > A, (p. Arg29Gln)) associated with the phenotype of oocyte death in a non-consanguineous family. This variant displayed an autosomal dominant inheritance pattern with reduced penetrance. Western blot analysis confirmed that the missense mutation of PANX1 (c.86G > A) altered the glycosylation pattern in HeLa cells. Moreover, the mutation effects on the function of PANX1 were weaker than recently reported variants. Our findings expand the inheritance pattern of PANX1 variants to an autosomal dominant mode with reduced penetrance and enrich the variational spectrum of PANX1. These results help us to better understand the genetic basis of female infertility with oocyte death.

Evaluation the expression of insulin and insulin receptor-beta (IR-β) in sperm of infertile male with failed intracytoplasmic sperm injection (ICSI)

Introduction: Most infertility treatment centers are currently using semen analysis to differentiate between fertile and infertile individuals. Recent studies have evaluated the expression and secretion of independent insulin and related beta receptor (IR-β) in mammals’ ejaculated spermatozoa such as humans and pigs, and its effect on motility parameters capacitation and acrosomal reaction. Objectives: This study aimed to investigate the expression of insulin, IR-β and the parameters of sperm and DNA fragmentation index (DFI) in men with intracytoplasmic sperm injections (ICSI) failed. Patients and Methods: This study was conducted on 15 men with unknown infertility with a history of unsuccessful ICSI and 10 fertile men. After collecting samples, sperm parameters (concentration and motility), sperm DNA fragmentation and expression of insulin and IR-β genes were investigated. To assess DFI from SDFA kit and to review the expression, the genes were analyzed via RT-PCR method. Data were analyzed statistically by t-test and covariance tests. Results: Based on the findings of this research, the concentration and percentage of motility in infertile men with a history of unsuccessful ICSI was significantly lower than fertile individuals, moreover, the amount of DNA fragmentation significantly increased in infertile men with a history of unsuccessful ICSI compared to fertile men (P &lt; 0.05). Insulin expression in the sperm of infertile men with a failed ICSI history was significantly lower than fertile men (P &lt; 0.05); however, the IR-β expression rate was not significantly different between the two groups. Conclusion: Evaluation of insulin gene expression can be helpful as a good marker for identifying infertility with an unknown cause.

TOTAL MOTILE SPERM COUNT IS ASSOCIATED WITH ICSI SUCCESS USING SPERM OBTAINED BY TESE

In vitro fertilization (IVF) used in combination with intracytoplasmic sperm injection (ICSI) is indicated in couples with male factor infertility. Testicular sperm extraction (TESE) may improve success rates of ICSI by isolating sperm that have been shielded from oxidative damage. To date, few studies have identified semen parameters that predict success for IVF/ICSI using TESE sperm, and the effect of total motile sperm count (TMSC) has not been described. We sought to determine the effect of TMSC on ICSI outcomes in couples with isolated male factor infertility that underwent TESE. Data was collected prospectively from couples undergoing ICSI for isolated non-azoospermic male factor infertility using TESE sperm retrieved from 1/2016-1/2019. Couples were compared according to TMSC ≤ or >10 million per ejaculate. Biochemical pregnancy rates were compared in the two groups using linear logistic regression, controlling for female partner age and BMI. Twenty three couples underwent ICSI using TESE sperm. Median male partner age was 36.9 years (IQR:31.1-51.6) and median TMSC per ejaculate was 3.9 million (IQR:1.0-12.1). Female partner median age was 32.0 years (IQR:29.1-34.9) and median BMI was 27 (IQRL24-37). Six of seven (85.7%) couples in which the male partner had TMSC > 10 million achieved pregnancy, versus 4/16 (25%) with TMSC ≤ 10 million (p=0.007). TMSC >10 million was predictive of biochemical pregnancy (OR 17.0, CI 1.48-196, p=0.023). TMSC ≤10 million per ejaculate is predictive of ICSI failure in couples with isolated non-azoospermic male infertility.

Hysteroscopic Endometrial Biopsy after Intracytoplasmic Sperm Injection Failure for women with Unexplained Infertility

Background: Hysteroscopy is one of the most significantexaminations in females with recurrent implantation failure (RIF). It permits dependable visual evaluation of the cervical canal and uterine space and it taken into consideration as a gold standard for diagnosing intra-uterine pathology and has slight intra-operative and postoperative morbidities. Objectives: This work aimed to clarify the roles of Hysteroscopy and endometrial biopsy in evaluation of endometrium after ICSI failure in females with un-explained Infertility. Patients and methods: This was a prospective cohort investigation carried out at outpatient infertility clinic of the Assisted Reproductive Unit, Obstetrics and Gynecology department, Qena University Hospital. Women attend to Assisted Reproductive unit planned for intracytoplasmic sperm injection (ICSI). Results: Based on histology as a reference standard, hysteroscopy identified the chronic endometritis pathology in 26 patients (true positives) that also found by histology. Our study revealed that Hysteroscopy had overall sensitivity, specificity, and diagnostic accuracy of 49.06%, 53.19% and 51% respectively in detecting the chronic endometritis in our patients with positive predictive value (PPV) was 54.17% while the negative predictive value (NPV) was 48.08%. Conclusion: Hysteroscopy is of a great value in the assessment and diagnosis of the endometrial pathology in patients with unexplained RIF. There is no consensus on the everyday use of hysteroscopy before ICSI; hence, the importance of hysteroscopy as a routine in the managing of non-fertile females still remains a subject of investigation.

Biallelic mutations in spermatogenesis and centriole-associated 1 like (SPATC1L) cause acephalic spermatozoa syndrome and male infertility.

Acephalic spermatozoa syndrome is a rare type of teratozoospermia that severely impairs the reproductive ability of male patients, and genetic defects have been recognized as the main cause of acephalic spermatozoa syndrome. Spermatogenesis and centriole-associated 1 like (SPATC1L) is indispensable for maintaining the integrity of sperm head-to-tail connections in mice, but its roles in human sperm and early embryonic development remain largely unknown. Herein, we conducted whole-exome sequencing (WES) of 22 infertile men with acephalic spermatozoa syndrome. An in silico analysis of the candidate variants was conducted, and WES data analysis was performed using another cohort consisting of 34 patients with acephalic spermatozoa syndrome and 25 control subjects with proven fertility. We identified biallelic mutations in SPATC1L (c.910C>T:p.Arg304Cys and c.994G>T:p.Glu332X) from a patient whose sperm displayed complete acephalia. Both SPATC1L variants are rare and deleterious. SPATC1L is mainly expressed at the head–tail junction of elongating spermatids. Plasmids containing pathogenic variants decreased the level of SPATC1L in vitro. Moreover, none of the patient's four attempts at intracytoplasmic sperm injection (ICSI) resulted in a transplantable embryo, which suggests that SPATC1L defects might affect early embryonic development. In conclusion, this study provides the first identification of SPATC1L as a novel gene for human acephalic spermatozoa syndrome. Furthermore, WES might be applied for patients with acephalic spermatozoa syndrome who exhibit reiterative ICSI failures.

Impact of thawing-loading interval of frozen embryo on pregnancy outcome: retrospective analysis

Embryo transfer is a critical final step in the in vitro fertilization (IVF) cycle. Much attention has been paid to standardizing the procedures of embryo transfer to improve pregnancy outcome. Objective. To identify the impact of embryo thawing-loading interval (the time interval passing from thawing to loading of frozen embryo into the catheter) on pregnancy outcome. Results. The study was divided into quartiles according to thawing-loading interval in minutes (G I: &lt;60; G II: 60–119; G III: 120–239; G IV: 240 and above). Numerical variables are presented as median and interquartile range, and inter-group differences are compared using the Mann-Whitney U test. Categorical variables are presented as numbers and percentages, and differences are compared using the chi-square test for trend. After adjustment for the age of women, time of previous failed ICSI (Intracytoplasmic sperm injection) trials, number and maturity of transferred embryos, a thawing-loading interval between 60–119 minutes (adjusted odds ratio = 2.222, 95% CI = 1.370 to 3.603, p-value = 0.001) or between 120–239 minutes (adjusted odds ratio = 1.924, 95% CI = 1.172 to 3.161, p-value = 0.010) was independently associated with increased probability of clinical pregnancy compared with a thawing-loading interval of &lt;60 minutes. On the other hand, a thawing-loading interval of ≥240 minutes significantly decreased the probability of clinical pregnancy (adjusted odds ratio = 0.378, 95% CI 0.215 to 0.664, p-value = 0.001). Conclusion. After adjusting for potential confounders, this analysis found that a thawing-loading interval between 60–239 minutes was independently associated with better pregnancy outcomes compared with a thawing-loading interval of &lt;60 minutes. On the other hand, a thawing-loading interval of ≥240 minutes was significantly associated with a worse outcome. Key words: embryo transfer, IVF (in vitro fertilization), frozen embryo, thawing-loading interval

Impact of changing the ovarian hyperstimulation protocol in the sequental cycle on ICSI outcomes in the same patient population

AimWhether to use the same ovarian stimulation protocol or change it after a failed in vitro fertilization cycle is still a controversial issue. In this study we aimed to investigate the effect of changing the stimulation protocol in the sequental cycle of the same patient population on intracytoplasmic sperm injection (ICSI) outcomes. MethodsThis retrospective study included two sequental fresh ICSI cycles of 941 infertile women attended in a period of one year. Group A was composed of patients who failed to have clinical pregnancy with a GnRH agonist and group B was composed of patients who failed to have clinical pregnancy with a GnRH antagonist protocol cycle. In both groups the study group was composed of patients whose stimulation protocol was changed in the sequential cycle and the control group was composed of patients who proceeded with the same stimulation protocol. The clinical pregnancy and live birth rates were primary outcomes. ResultsIn group A, the clinical pregnancy rates were comparable between the study and the control groups, but the live birth rate was higher in the study group (p=0.03). In group B, there was no difference in terms of clinical pregnancy and live birth rates between the study and control groups (p=0.740 and p=0.842 respectively). ConclusionsChanging the ovarian stimulation protocol after a failed ICSI cycle downregulated with a GnRH agonist increased the live birth rate. After a failed cycle stimulated with a GnRH antagonist protocol, protocol change did not have any impact on the clinical pregnancy and live birth rates.

Reproductive outcomes with donor sperm in couples with severe male-factor infertility after intracytoplasmic sperm injection failures.

To evaluate reproductive outcomes of artificial insemination and IVF with donor sperm (AID or IVF-D) for male-factor couples with a history of unsuccessful ICSI attempt. This retrospective cohort includes couples with severe male-factor infertility who failed ICSI treatment, and subsequently underwent semen donation treatment. We report the following outcomes: (1) live birth rates in AID and IVF-D treatment for couples with severe male infertility factors and prior ICSI failures; (2) paternal impact on embryo development of the same oocyte cohort; (3) prognostic factors in obtaining a live birth with donor semen. Of 92 women with failed ICSI cycles (26 with multiple attempts), 45 couples underwent AID treatment. Live birth rate per cycle of AID was 18.9%. Fifty-three patients underwent IVF-D including 6 couples who previously did not conceive with AID. Embryological outcomes including fertilization, viable cleavage embryos, and blastocyst formation rates were significantly lower in ICSI cycles with partner sperm compared with IVF-D (P < 0.01). Logistic regression analysis showed that female age and the severity of spermatogenetic disorder are prognostic factors in obtaining a live birth with donated sperm. Couples with severe male infertility factor (azoospermia or extreme oligoasthenospermia) and a history of unsuccessful ICSI cycles benefit from treating with donor sperm. ICSI fertilization, embryo viability, and progression of the embryo to the blastocyst stage are significantly deteriorated by semen parameters. The prognostic factors identified may help couples plan their treatment and prepare for their parenthood journey.

Sperm Genomic Integrity by TUNEL Varies throughout the Male Genital Tract.

We assessed sperm chromatin fragmentation at different levels of the male genital tract. Ejaculated specimens from consenting male partners were screened for sperm chromatin fragmentation by TUNEL (terminal deoxynucleotidyl deoxyuridine triphosphate nick end labeling). Men with intracytoplasmic sperm injection failure and high ejaculated sperm chromatin fragmentation underwent surgery to retrieve spermatozoa from different levels of the male genital tract, which were then reassessed for sperm chromatin fragmentation. Approximately 500 or more spermatozoa were assessed per patient with a 15% threshold. Intracytoplasmic sperm injection results of cycles using spermatozoa from different levels of the male genital tract were compared. Topographical assessment of the male genital tract showed a mean ± SD of 20.4% ± 10% sperm chromatin fragmentation in the vas deferens, 15.8% ± 8% in the epididymis and 11.4% ± 6% in the testis. All values were lower than in ejaculated controls (mean 32.9% ± 20%, p <0.05). A total of 25 couples who underwent intracytoplasmic sperm injection with surgically retrieved spermatozoa had lower sperm chromatin fragmentation (p <0.001), and higher implantation, clinical pregnancy and delivery rates (p <0.01). A total of 45 couples with a history of intracytoplasmic sperm injection failure with ejaculate performed elsewhere were treated solely with surgically retrieved spermatozoa at our center. Compared to historical cycles, surgically retrieved spermatozoa had a lower fertilization rate (65%, p <0.05) but enhanced rates of implantation (19.1%), clinical pregnancy (40.0%) and delivery (34.3%) (each p <0.01). To our knowledge we report for the first time that sperm chromatin fragmentation increases progressively from the testicle to the epididymis and the vas deferens, and is highest in the ejaculate. Men with high ejaculated sperm chromatin fragmentation can benefit from using surgically retrieved sperm for in vitro fertilization and/or intracytoplasmic sperm injection.