Intracytoplasmic Sperm Injection Fertilization Research Articles

Microinjection manipulation decreases the expression of GABA-A receptor signaling pathway genes in mouse embryos derived using intracytoplasmic sperm injection.

BackgroundThe GABA‐A receptor signaling pathway regulates proliferation, differentiation, apoptosis, and responses to overt DNA damage during embryonic development.MethodsTo analyze the gene expression after intracytoplasmic sperm injection (ICSI) and in in vivo mouse embryos, the global pattern of gene expression dataset, GSE23009, was obtained from the Gene Expression Omnibus database. Genes with differential expression were identified using the R software package, and RT‐qPCR was performed to confirm the microarray results.ResultsMouse blastocysts derived from ICSI fertilization had decreased expression of GABA‐A receptor signaling pathway genes. However, the mechanisms underlying these changes were not elucidated. The gene expression of the GABA‐A pathway was not significantly different between blastocysts obtained from IVF and in vivo fertilization. However, microinjection after IVF significantly reduced the expression of the GABA‐A pathway gene to levels similar to those in the ICSI group.ConclusionBased on our results, decreased gene expression is a result of the microinjection manipulation performed during ICSI.

Comparison of aneuploidy rates between conventional in vitro fertilization and intracytoplasmic sperm injection in in vitro fertilization–intracytoplasmic sperm injection split insemination cycles

ObjectiveTo evaluate the influence of insemination methods on outcomes of preimplantation genetic testing for aneuploidy (PGT-A) by assessing PGT-A results in embryos that derived from conventional in vitro fertilization (IVF) versus intracytoplasmic sperm injection (ICSI) in sibling oocytes.DesignRetrospective cohort study.SettingSingle academic IVF center.Patient(s)A total of 118 couples who underwent 131 split insemination cycles from July 2016–July 2019.Intervention(s)In all cycles, sibling oocytes were allocated randomly to conventional IVF or ICSI prior to stripping. Preimplantation genetic testing for aneuploidy was performed via trophectoderm biopsy and next-generation sequencing with 24-chromosome screening.Main Outcome Measure(s)Rates of euploid, aneuploid, and mosaic embryos per biopsy.Result(s)A total of 2,129 oocytes were randomized to conventional IVF (n = 1,026) and ICSI (n = 1,103). No difference was observed in the aneuploidy rates (50.3% vs. 45.2%) and percentages of mosaic embryos (1.7% vs. 2.4%) per biopsy between conventional IVF and ICSI sibling oocytes. Percentages of different aneuploidy types and aneuploidies that involved sex chromosome abnormalities (6.2% vs. 7.2%) were similar between the two groups. In the end, the overall chance to have an euploid embryo per allocated oocyte in the two groups was similar (13.2% vs. 15.5%).Conclusion(s)Blastocysts created with conventional IVF and ICSI using sibling oocytes had similar rates of aneuploidy and mosaicism as examined using 24-chromosome screening. It is unlikely that conventional IVF caused significant contamination during PGT-A. We recommend conventional IVF as the preferred insemination method in PGT-A cycles, and ICSI should be indicated only in cases of male-factor infertility.

The impact of late follicular progesterone level on in vitro fertilization-intracytoplasmic sperm injection outcome: Case-control study.

BackgroundStudies have been conducted to improve the pregnancy rate through the in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) program. In recent years, researchers have been focusing on finding impact of high progesterone level on endometrial receptivity. However, data on whether progesterone level also affects the quality of the embryo is still limited.ObjectiveThe aim is to assess the effect of late follicular progesterone level on the outcome of in vitro fertilization-intracytoplasmic sperm injection (IVF-ICSI).Materials and MethodsThis was a case-control of 245 women who underwent in vitro fertilization cycle at Halim Fertility Center, Indonesia. The outcomes assessed were number of oocytes retrieved (OR), maturation rate (MR), fertilization rate (FR), number of good embryos (GE), number of fair embryos (FE), and number of poor embryos (PE). The progesterone (P4) and estradiol (E2) levels were analyzed on the day of human chorionic gonadotropin injection. Serum progesterone level was divided into three groups: 1. low progesterone ( 0.50 ng/ml), 2. normal progesterone (0.51-1.50 ng/ml), and 3. high progesterone ( 1.50 ng/ml). All outcomes were compared amongst the groups.ResultsSignificant differences occurred between progesterone level on the day of human chorionic gonadotropin administration. The number of OR in group 1, 2, and 3 were 8.41 5.88 vs. 12.99 8.51 vs. 17.58 9.52, respectively.ConclusionProgesterone level on the day of human chorionic gonadotropin injection may have an impact on the outcome of IVF-ICSI.

The vaginal microbiome as a tool to predict IVF success.

Herein, we present an overview of the recent microbiome research and findings within the field of reproductive medicine and its relation with the outcome of Assisted reproductive technology (ART). Analyses of the microbiome composition all throughout the female reproductive trace during the process of assisted reproductive techniques are discussed. Only the vaginal microbiome can be sampled without possible risks of contamination. Although this also seems to apply to the cervical microbiome, collection has to be performed with extreme caution. Because of the high risk of cross contamination, results of microbial composition of all other sites of the female reproductive tract have to be interpreted with caution. The vaginal composition prior to the start of hormonal treatment for ART seems to be predictive of in vitro fertilization/in vitro fertilization-intracytoplasmic sperm injection (IVF/IVF-ICSI) outcome, with mainly a highly negative predictive value. The local microbiota, especially the absence or presence of specific microbes, within parts of the female reproductive tract seem to be associated with the outcome of ART.

Endometrial thickness is associated with incidence of small-for-gestational-age infants in fresh in vitro fertilization–intracytoplasmic sperm injection and embryo transfer cycles

To investigate whether endometrial thickness (EMT) is associated with adverse obstetric and neonatal outcomes in fresh invitro fertilization/intracytoplasmic sperm injection-embryo transfer (IVF/ICSI-ET) cycles. Retrospective cohort study. University-based reproductive medical center. Women under the age of 42 years who underwent IVF/ICSI treatment and received fresh ET in our unit from January 2017 to December 2018, resulting in a live singleton birth. Controlled ovarian hyperstimulation and IVF/ICSI; fresh ET. Birth weight, gestational age, small for gestational age (SGA), large for gestational age (LGA), placenta previa, placental abruption, hypertensive disorders, and gestational diabetes mellitus. The risk of being born SGA was statistically significantly increased in the EMT ≤7.5 mm group compared with those from the EMT >12 mm group (adjusted odds ratio [aOR] 2.391; 95% confidence interval [CI], 1.155-4.950). Moreover, maternal body mass index, secondary infertility, preterm delivery, and hypertensive disorders were all independent predictors for SGA. The mean birth weights of singletons in women with EMT ≤7.5 mm were lower than in the groups with EMT >7.5-12 mm and EMT >12 mm (3.25 ± 0.56 kg vs. 3.38 ± 0.51 kg and 3.39 ± 0.53 kg, respectively). After fresh IVF/ICSI-ET, the risk of SGA was increased twofold in women with EMT ≤7.5 mm compared with women with EMT >12 mm. We suggest that women with a thin EMT after obtaining a pregnancy by IVF should receive improved prenatal care to reduce the risk of delivering a SGA infant.

Effect of vitrification of in vitro matured prepubertal goat oocytes on embryo development after parthenogenic activation and intracytoplasmic sperm injection

This work studies the effect of vitrification of in vitro matured (IVM) prepubertal goat oocytes on: 1) oocyte damage assessed by reactive oxygen species (ROS) level and apoptosis and 2) embryo development after Intracytoplasmic sperm injection (ICSI) and Parthenogenic Activation (PA). Oocytes were IVM in supplemented TCM-199 for 22–24 h. Control group oocytes matured during 24 h were directly used for the analysis after IVM. Vitrified/warmed IVM-oocytes were vitrified after 22 h of IVM in 15% ethylene glycol (EG), 15% dimethyl sulfoxide (Me2SO) and 0.5 M sucrose and after subjected to warming procedure. Oocyte ROS level was measured by staining denuded IVM-oocytes with 10 μM 2′7′ dichlorodihydrofluorescein diacetate. Apoptosis was analyzed by Annexin V (AV) Apoptosis Detection kit and Propidium iodide (PI) signal and oocytes were classified as: Live (AV− PI−), early apoptotic (AV+ PI−), dead non-apoptotic (AV− PI+) and necrotic (AV+ PI+). Developmental competence of vitrified/warmed oocytes was assessed by PA (5 min in 5 μM Ionomycin plus 4 h in 2 mM 6-Dimethylaminopurine), and by ICSI fertilization. Presumptive zygotes were in vitro cultured for 8 days in commercial media BO-IVC. Vitrified/warmed oocytes showed higher ROS levels (P < 0.0001), lower live oocytes (44 vs. 66%; P: 0.0025) and higher dead non-apoptotic oocytes (33 vs. 13% P: 0.023) compared to control. No differences were found on normal zygote formation (2 PN) (32 vs. 25%) or blastocyst development (0 vs. 4%) after ICSI fertilization. However, after PA, significant differences were found in cleavage rate (59 vs.78%; P < 0.0343) and blastocyst formation (1 vs. 25%; P < 0.0001). In conclusion, vitrification reduced oocyte competence by increasing dead oocytes and ROS levels.

Chromosomal polymorphisms are associated with blastomere multinucleation in IVF/ICSI cycles

To investigate the association of chromosomal polymorphisms with multinucleated embryos in infertile couples undergoing in vitro fertilization-intracytoplasmic sperm injection (IVF-ICSI). This retrospective case-control study was conducted among 1145 infertile couples undergoing their first IVF/ICSI cycles. According to their karyotype, the couples were divided into chromosomal polymorphism group and control group, and the former group was divided into 3 subgroups: inversion group, D and G genome polymorphic group and 1, 9, and 16 qh+group. The blastomere multinucleation rate, clinical pregnancy rate and live birth rate were compared between the groups. Of the total of 1145 couples, 139 (6.10%) had chromosomal polymorphisms at least in one partner. No significant differences were found in female age, BMI, basal FSH level, total gonadotropin dose, E2 level on day of HCG, number of oocytes retrieved, fertilization rate, top quality embryo rate, clinical pregnancy rate or live birth rate among the groups (P > 0.05). The multinuclear rate of the embryos in couples with pericentric inversion of chromosomes 1, 9, and Y chromosomes and those with D and G genome polymorphisms were 8.23% and 4.65%, respectively, significantly higher than that in the control group (2.69%; P < 0.05); the multinuclear rate of the embryos was 2.77% in 1, 9, and 16 qh+ group, similar with that in the control group (P > 0.05). Infertile couples with pericentric inversion of chromosomes 1, 9, and Y chromosomes and in those with D and G genome polymorphism are at higher risks of blastomere multinucleation in IVF- ICSI cycles; 1, 9, and 16 qh + polymorphisms do not increase the rate of blastomere multinucleation of the embryos.

Relevance of Split In vitro Fertilization-Intracytoplasmic Sperm Injection Method of Insemination in Normozoospermic and Mildly Oligospermic Men: A Retrospective Study.

Background:Use of intracytoplasmic sperm injection (ICSI) is generally considered redundant in cases of normozoospermia, or mild male factor cases of infertility and conventional method of insemination is advocated. However, there is a risk of low fertilization or total fertilization failure (TFF) and to avoid this, split in vitro fertilization (IVF)-ICSI method of insemination is advised. In our study, we have shown that not only TFF is avoided with split method of insemination, but also cancellation of embryo transfer (ET) can be avoided in a significant number of IVF cycles.Aims:This study aimed to assess whether the IVF-ICSI split insemination method was able to reduce the risk of ET cancellation in couples with normal or mild sperm characteristics.Settings and Design:It is a retrospective study including a total of 107 split insemination cycles done at our center.Materials and Methods:The female partner's age was under 37 years, and at least ten oocytes were retrieved in all cycles. Sibling oocytes were randomly allocated to IVF or ICSI. Statistical analysis was carried out using Graphpad Prism, Instat.Results and Conclusion:The fertilization rate in oocytes kept in conventional IVF was significantly higher (79.8%) compared to that of oocytes injected through ICSI (69.1%). Only one couple had TFF. In majority of the cycles, i.e., 97 out of 107 cycles, the mode of insemination did not affect the fertilization rate or embryo quality. Nearly 28% of the cycles were saved from ET cancellation by adopting the split insemination method. “Split IVF-ICSI” approach can save a significant number of ART cycles and is found to be cost-effective as it avoids incurring the cost of two ART cycles.

Predictors of blastocyst formation rate in elective day 5 transfer cycle

This study aimed to investigate the factors affecting blastocyst formation rate. One hundred and seven fresh in vitro fertilisation (IVF) and elective day 5 blastocyst transfer cycles were selected. Univariate and multivariate analyses revealed that intracytoplasmic sperm injection (ICSI) (r = −.236, p = .014 vs. p = .005) was advantageous for blastocyst formation. In addition, the number of mature oocytes (r = −.274, p = .004 vs. p = .002) was a significant factor associated with blastocyst and good-quality blastocyst formation rates (p = .021, r = −.389). Both blastocyst and good-quality blastocyst formation rates were significantly higher with ICSI than with conventional insemination (65.0 ± 24.5% vs. 50.0 ± 21.2%, p = .012; 43.1 ± 22.8% vs. 30.9 ± 19.8%, p = .038, respectively). The number of mature oocytes appears to be the most important predictor of blastocyst formation rate. Additionally, ICSI fertilisation is superior to conventional insemination in terms of blastocyst formation rate.IMPACT STATEMENTWhat is already known on this subject? There are many advantages of blastocyst transfer cycle over cleavage transfer cycle, but there are no known routine selection criteria for the timing of embryo transfer. To date, the number of blastomeres, number of retrieved oocytes, quality of embryos and fertilisation method have been suggested as the important factors involved in blastocyst formation. However, the number of studies on this issue is limited, and some studies have shown conflicting results.What do the results of this study add? This study showed that the number of mature oocytes and ICSI fertilisation are the significant factors associated with blastocyst formation rate in elective day 5 transfer cycle.What are the implications of these findings for clinical practice and/or further research? This paper demonstrated that the number of mature oocytes and the fertilisation method should be considered before embryo transfer. Consideration of these factors would be meaningful in selecting patients who will be suitable for extended culture up to day 5.

201 Correlation between metaphase II oocyte cytoplasmic granulation patterns and intracytoplasmic sperm injection fertilization outcome in older patients

Cytoplasmic granulation is frequently observed in human MII oocytes in fertility clinics from women requiring early egg retrieval (also called highly individualized egg retrieval, with hCG trigger at lead follicle size 16.0-18.0mm or even less) to avoid premature luteinization. Early retrieval is mostly done for older women (≥36 years old) or younger women with premature ovarian aging. However, only limited reports have focused on detailed analysis of the patterns of cytoplasmic granulation and their correlation with the quality of human oocytes. The aim of this report was to evaluate correlation between granulation patterns of human MII oocytes from early retrieval and the corresponding fertilization rates from intracytoplasmic sperm injection (ICSI). Each MII oocyte was imaged when ICSI was performed. The granulation pattern of each oocyte was reviewed, evaluated, and categorized by the same person as non-granulation (NG, homogeneous, sandy pattern, no granulation or granulation area &amp;lt;1/8 of total cytoplasm), central granulation (CG, ring pattern, a cluster of granules in centre with size &amp;gt;1/4 of oocyte diameter), uneven granulation (UG, sandy-rocky pattern, granulation area &amp;gt;1/8 but &amp;lt;3/4 of total cytoplasm), and dispersed granulation (DG, rocky pattern, granulation area &amp;gt;3/4 of total cytoplasm). Fertilization (pronucleus count, PN) results were then extracted from medical records and matched to the granulation categories. In total, 1759 oocytes done with ICSI in year 2018 were analysed. Fertilization (&amp;lt;2PN, 2PN or &amp;gt;2PN) rates of each group (n=381, n=1222, n=156, respectively) were calculated. Student's t-test was used for group-group comparison analysis. Compared with the NG group, the UG and CG groups had lower 2PN rates (67.2% vs. 84.6%; P&amp;lt;0.001; 73.0% vs. 84.6%; P&amp;lt;0.05; respectively) and the DG group had a much lower 2PN rate (43.3% vs. 84.6%; P&amp;lt;0.001). The UG and CG groups had higher &amp;lt;2PN rates (24.8% vs. 12.1%; P&amp;lt;0.001; 24.3% vs. 12.1%; P&amp;lt;0.001; respectively), and the DG group had a much higher &amp;lt;2PN rate (41.1% vs. 12.1%; P&amp;lt;0.001) than the NG group. For &amp;gt;2PN rates, both UG and DG groups were higher than the NG group (8.0% vs. 3.3%; P&amp;lt;0.05; 15.6% vs. 3.3%; P&amp;lt;0.001; respectively), whereas the DG group was higher than the UG group (15.6% vs. 8.0%). These data demonstrated that dispersed granulation was the worst pattern for fertilization. Interestingly, many of the uneven-granulation group oocytes could become homogeneous (non-granulation) after hours or overnight IVM culture, suggesting that uneven granulation might be a sign of incomplete cytoplasmic maturity. Technically, performing ICSI through the less-granulation side was easier because of less resistance to membrane breaking and to pushing sperm out. Discovering the molecular nature of the granules and related cytoskeleton structures would extend our understanding of their existence and improve treatments by adjusting stimulation and trigger accordingly.

Could fertility clinics offer a sizable improvement of live birth rates by maturing post-GVBD oocytes in vitro?

The objective of this study is to analyze the potential of immature, denuded, post-GVBD (germinal vesicle breakdown) oocytes (including prometaphase I, metaphase I, and prometaphase II stages) to result live birth after in vitro maturation. Furthermore, we compared two culture media to identify which of them provides better reproductive outcomes when used for in vitro maturation. We performed a retrospective cohort study including 4022 IVF-ICSI (in vitro fertilization-intracytoplasmic sperm injection) cycles between 2011 and 2015. A total of 4450 immature post-GVBD oocytes from 1442 cycles were cultured in vitro; of these, 2364 oocytes reached MII (metaphase II) stage (IVMC oocytes, in vitro meiotic completion) and were fertilized. Overall, 3933 embryo transfers were performed: 3579 were embryos derived from MII oocytes (ET-MII); 264 were embryos derived from MII + IVMC oocytes (ET-MIX), and 90 embryos from IVMC oocytes (ET-IVMC). In total, 399 IVMC embryos were transferred. Maturation rate for immature post-GVBD oocytes was 54.1%. G-2™PLUS (Vitrolife) medium provided significantly higher maturation rate (p < 0.001) than G-IVF™PLUS (Vitrolife) (65.7% vs. 42.5%, p < 0.001). Embryos in ET-IVMC in cleavage stage had an average morphological score of 6.8/10 (7.7 in ET-MII; p < 0.001). Regarding reproductive outcomes, ET-IVMC gave 11.1% biochemical pregnancy rate, 10.0% clinical pregnancy rate, 7.8% ongoing pregnancy rate, and 5.6% live birth rate. Embryos arising from IVMC oocytes resulted in a live birth rate of 5.6%. We suggest that in vitro maturation of denuded immature post-GVBD oocytes should be performed at the very least when few MII are collected, and likely in all patients, as they provide acceptable maturation and fertilization rates, and a sizeable increase in live birth.

Artificial oocyte activation improves cycles with prospects of ICSI fertilization failure: a sibling oocyte control study.

Does artificial oocyte activation improve clinical outcomes for patients at risk of intracytoplasmic sperm injection (ICSI) fertilization failure? In this study, sibling oocytes from patients with previous ICSI failure or severe teratozoospermia were divided equally into two groups, half for artificial oocyte activation (AOA) with ionomycin after conventional ICSI and the other half for conventional ICSI only (non-AOA). The fertilization rates, cleavage rates, transferable embryo rates and blastulation rates of the two groups were compared first; the clinical pregnancy and live birth rates were also compared to assess the efficiency and safety of AOA. The outcomes of the AOA group were significantly better than those of the conventional ICSI group in terms of the fertilization (50.38% versus 33.86%, respectively, P<0.001), cleavage (59.16% versus 39.04%, respectively, P<0.001) and transferable embryo rates (43.51% versus 26.69%, respectively, P<0.001). The blastulation (43.53% versus 36.11%, respectively), implantation (26.83% versus 15.79%, respectively), clinical pregnancy (38.46% versus 25%, respectively) and live birth rates (38.46% versus 16.67%, respectively) were not significantly different. This study showed that AOA improved some aspects of cycles at risk of ICSI failure by increasing the fertilization and transferable embryo rates. But blastulation, pregnancy and implantation rates were not improved. The study is limited by its small size and absence of data on cumulative outcomes.

Messenger RNA and protein expression of tumor necrosis factor α and its receptors in human follicular granulosa cells.

To evaluate the concentration of tumor necrosis factor α (TNF-α) and its soluble receptors (sTNFR I and II) in serum and follicular fluid (FF) at the time of oocyte retrieval and to detect expression of TNF-α and its receptors by luteinized granulosa cells (GCs). In a cross-sectional study and through an in vitro fertilization-intracytoplasmic sperm injection (IVF-ICSI) program, 81 women undergoing oocyte retrieval were recruited. Serum and FF were obtained from 81 women. GCs were pooled from 20 patients (from six different days of oocyte retrievals, 5-16 follicles per patient). TNF-α and its soluble receptors concentration were determined by enzyme-linked immunosorbent assay and also their expression by immune cytochemistry and reverse-transcription polymerase chain reaction analysis. The median TNF-α concentration in serum was 4.06 pg/ml (interquartile range [IQR], 3.71-6.14) and significantly higher than that in FF with 3.50 pg/ml (IQR, 3.05-5.01), p < 0.001. The sTNFR I and II levels in serum were lower and higher than FF, respectively. The TNF-α levels in serum and FF of good responders were higher than low responders (p = 0.017 and 0.021, respectively). TNF-α cut-off level for low responders versus good responders was 4.174 pg/ml in serum with a pregnancy rate of 25.8% and 40% for below and above of this level, respectively (p = 0.19). For FF, the cut-off value was 3.89 pg/ml. TNF-α and its receptors were expressed by GCs. The presence of TNF-α and its soluble receptors in serum and FF and their expression by GCs suggest an important role for this cytokine in ovarian function.

Zeta method: A noninvasive method based on membrane charge for selecting spermatozoa expressing high level of phospholipaseCζ.

Intracytoplasmic sperm injection (ICSI) is a technique developed to help attain successful fertilisation for couples with severe male factor. However, a small percentage of couples confront low or failed fertilisation, mainly due to failed oocyte activation. Several studies have introduced phospholipase Cζ (PLCζ) as the main sperm factor inducing oocyte activation and thereby has the potential to act as a biomarker for the prediction of ICSI fertilisation outcome. On the other hand, researchers have focused on novel sperm selection procedures based on cellular characteristics of spermatozoa such as surface electrical charge (Zeta potential) to isolate normal sperm subpopulation with intact chromatin. Therefore, we aimed to compare PLCζ between Zeta method and routine sperm preparation procedure: density gradient centrifugation (DGC). Our results showed that number of PLCζ-positive spermatozoa was significantly low in the Zeta method, but the intensity of PLCζ protein in such spermatozoa was significantly higher than DGC procedure. Therefore, the combination of DGC with Zeta procedure may allow selecting the population of spermatozoa with a high percentage of PLCζ which may also contain a high amount of PLCζ and with intact chromatin. This sperm selection procedure can open a new approach for infertile men with previously failed fertilisation.

A point of confusion for embryologists in the identification of viable spermatozoa by the eosin-nigrosin test.

A viable spermatozoon is a prerequisite for fertilization in intracytoplasmic sperm injection (ICSI). Thus, it is crucial to select viable but immotile spermatozoa on the day of ICSI. We report conflicting results in the identification of viable but immotile spermatozoa between the eosin-nigrosin staining and the laser test, which resulted in confusion for embryologists during assisted reproductive technology (ART). Three patients’ semen samples that showed no motile spermatozoa are described in this report. To identify viable spermatozoa, we used both the eosin-nigrosin test and the laser test for each sample, and repeated the semen analysis twice in each patient. Viable but immotile spermatozoa selected by the laser test were used for ICSI. Viable spermatozoa were detected by both the eosin-nigrosin and laser tests in two patients (case 1, 95.00% vs. 24.21% and 92.68% vs. 22.22%; case 2, 41.18% vs. 23.48% and 39.81% vs. 22.52%), indicating consistent results between the two methods. In the third patient, the eosin-nigrosin test yielded viability rates of 20.75% and 19.14%, while the result of the laser test was 0%. Thus, testicular aspiration was performed to collect viable sperm from this patient. Normal fertilization was achieved after the injection of viable but immotile spermatozoa selected from these patients by the laser test, resulting in the birth of two healthy babies. Our study documents a case where the eosin-nigrosin test showed a limitation in identifying viable but immotile spermatozoa for ART, while the laser test may overcome this limitation. Larger samples may be required to corroborate the clinical value of the laser test.

The Effect of Sperm DNA Fragmentation Index on the Outcome of Intrauterine Insemination and Intracytoplasmic Sperm Injection.

Context:Elevated sperm DNA fragmentation index (DFI) is found to affect normal embryonic development, implantation and fetal development after intrauterine insemination (IUI), in vitro fertilization and intracytoplasmic sperm injection (ICSI). Estimation of DFI by terminal deoxynucleotidyl transferase-mediated fluorescent deoxy uridine nucleotide nick end labeling (TUNEL) assay was found to have a high predictive value for pregnancy after fertility treatments.Aim:This study aims to find the effect of increased sperm DNA fragmentation by TUNEL assay on reproductive outcome after IUI and ICSI.Primary Objective:To find the association of DFI and pregnancy rate in IUI and ICSI.Secondary Objectives:To find the association of DFI with fertilization and implantation in ICSI. To find the association of DFI with miscarriage rate.Settings and Design:A prospective observational study performed at a tertiary care university teaching hospital.Subjects and Methods:105 male partners of infertile couple planned for IUI and ICSI underwent estimation of sperm-DFI by TUNEL assay. The treatment outcomes were compared between the DFI-positive (≥20%) and DFI-negative (<20%) groups.Statistical Analysis Used:Statistical analysis was performed by SPSS version 17, Software. P < 0.05 was considered statistically significant.Results:The men with abnormal semen analysis were significantly higher in the DFI-positive group (77.15% vs. 22.85%). There was no significant difference in the pregnancy rate in IUI cycles (17.6% vs. 11.8%); but in the ICSI, the pregnancy rate was significantly reduced in the DFI-positive group (16.7% vs. 47.4%).Conclusions:Elevated DFI significantly affects the pregnancy rate in ICSI cycles.

Anxiety and Stress at Different Stages of Treatment in Women Undergoing In vitro Fertilization-Intracytoplasmic Sperm Injection.

Aim:The aim of the present study is to evaluate the state anxiety (the present state of mind), trait anxiety (general anxiety), as well as perceived stress in women undergoing in vitro fertilization (IVF) treatment at three stages: T1 (on the day of start of stimulation), T2 (on the day of embryo transfer), and T3 (10 days after embryo transfer). The data at T3 level were collected telephonically.Methodology:The present study was carried out on 137 women undergoing IVF intracytoplasmic sperm injection cycle at four different clinics of four cities from October to April 2016. State-trait anxiety inventory (Spielberger) and perceived stress scale (Okun, et al.) were used as the tools.Statistical Analysis:The analysis was done at two levels; descriptive and inferential (analysis of variance [ANOVA], Student's t-test, Levene's test) using SPSS v16.Results:The state anxiety was higher at all the three levels than trait anxiety. The overt anxiety was highest at T3 level (mean = 45.77) followed by T1 level (mean = 44.23) and T2 level (mean = 43.04). Perceived stress was elevated at T1 level (mean = 17.93) followed by T3 level (mean = 17.28) and T2 level (mean = 16.72). The results of ANOVA showed a significant difference in anxiety among all the three levels (P = 0.036), but no significant difference was found for perceived stress (P = 0.169). t-test revealed that there was a significant difference between state and trait anxiety at T1, T2, and T3 levels (P = 0.01, P = 0.21, P = 0.00, respectively). A significant difference was only seen between the T1 and T2 levels in perceived stress (P = 0.052). In state anxiety, a significant difference was observed only between T2 and T3 levels (P = 0.23).Conclusion:It was observed that anxiety and stress are present in women throughout the treatment. The waiting period (T3) is the most anxious for them and their level of state anxiety is higher compared to their trait anxiety. Perceived stress is observed to be more on the day of start of stimulation followed by the waiting period.

Establishing an Anti-Müllerian Hormone Cutoff for Diagnosis of Polycystic Ovarian Syndrome in Women of Reproductive Age-Bearing Indian Ethnicity Using the Automated Anti-Müllerian Hormone Assay.

Context:Polycystic ovary syndrome diagnosed by Rotterdam criteria, is the most common cause of anovulatory infertility. The criteria of polycystic ovarian morphology (PCOM) are subject to operator variability and technological advances. Serum anti-Müllerian hormone (AMH) level has been proposed as a more reliable alternative to antral follicle count. There is a paucity of data on use of AMH for diagnosis of PCOS in Indian women.Aim and Objectives:The aim of this study is to determine a cutoff level for AMH that could facilitate diagnosis of PCOS and its phenotypes in women of Indian origin using the automated (Roche) assay and to compare the competence of oocytes in PCOS and non-PCOS women undergoing in vitro fertilization-intracytoplasmic sperm injection (IVF-ICSI).Materials and Methodology:A total of 367 women undergoing treatment at our fertility center between February 2017 and August 2017 were prospectively enrolled in this study. Of these, 133 were diagnosed with PCOS, 69 had isolated PCOM, and 165 (controls) had normal ovaries on ultrasound examination. Serum AMH levels were assessed using the fully automated Roche Elecsys® immunoassay. Gonadotropin-releasing hormone antagonist protocol was used for IVF-ICSI in all patients.Statistical Analysis Used:Quantitative variables were compared using the Mann–Whitney test. Qualitative variables were correlated using the Chi-square test. P < 0.05 was considered to be statistically significant.Results:Mean AMH concentrations in women with PCOS was higher (7.56 ± 4.36 ng/mL) in comparison to PCOM and controls. Serum AMH concentration >5.03 ng/mL could facilitate diagnosis of PCOS (area under the curve = 0.826); sensitivity –70.68%, specificity of 79.91%. There was no difference in the ratio of mature to total oocytes retrieved in the three groups (P > 0.05). Mean number of mature oocytes was lower in controls than PCOS and PCOM (P < 0.001).Conclusions:Serum AMH concentration >5.03 ng/mL could be used as cutoff value for the diagnosis of PCOS in women of Indian origin.

Analysis of two different luteal phase support regimes and evaluation of in vitro fertilization-intra cytoplasmic sperm injection outcomes.

Objective:To evaluate clinical pregnancy rates, miscarriage rates, ongoing pregnancy rates, and in vitro fertilization-intra cytoplasmic sperm injection outcomes of gonadotropin releasing hormone agonist (GnRHa) administration compared with human chorionic gonadotropin (hCG) application for luteal phase support.Materials and Methods:A total of 456 patients were included in the study. The patients were divided into two groups according to luteal phase support type: in group 1 (n=158), single-dose triptorelin acetate 0.1 mg was given on the sixth day after the oocyte pick-up (OPU). In group 2 (n=298), hCG 1500 IU was given on day 4, 7 and 10 after the OPU.Results:Both groups were homogeneous in relation with age and antral follicle count. The number of stimulation days and endometrial thickness on hCG day (mm) were found to be significantly higher in group 2 than in group 1 (p<0.001). The clinical pregnancy rate was slightly higher in the GnRHa group, but this difference was not statistically significant.Conclusion:Although there was no statistically significant difference between the two groups, luteal phase support with single-dose GnRHa might be as efficient as three doses of hCG. Large prospective, randomized-controlled studies are required comparing GnRHa and hCG for luteal phase support.

Effect of type 3 intramural fibroids on in vitro fertilization–intracytoplasmic sperm injection outcomes: a retrospective cohort study

To evaluate the influence of type 3 intramural fibroids on invitro fertilization-intracytoplasmic sperm injection (IVF-ICSI) outcomes. Retrospective cohort study. University-based reproductive medicine center. All women undergoing IVF-ICSI from January 1, 2009, to December 31, 2016, in our unit. Each woman was matched with three separate control subjects of the same age (±1year), number of cycles, type of infertility (primary or secondary), type of protocol used for controlled ovarian hyperstimulation (COH), and no uterine fibroids identified by transvaginal ultrasound. Implantation, clinical pregnancy, clinical miscarriage, and live birth rates. We included 151 patients with type 3 intramural fibroids and 453 matched control subjects who underwent IVF-ICSI. The rate of "other protocol" used in COH was significantly higher in women with type 3 fibroids than in the control subjects (P<.001). The experimental group had a significantly lower implantation rate. Type 3 fibroids also resulted in a lower frequency of live births and clinical pregnancy. There was no significant difference between the groups in the rate of clinical miscarriage. Compared with the corresponding control subjects, patients with type 3 fibroids with a single fibroid diameter (SD) or total reported fibroid diameter (TD) >2.0cm also had significantly lower rates of live birth, clinical pregnancy, and implantation. Type 3 fibroids with SD or TD ≤2.0cm had no significant difference in IVF-ICSI outcomes compared with corresponding control subjects. Our results suggest that type 3 fibroids exert a negative impact on the rates of implantation, clinical pregnancy, and live birth in patients undergoing IVF-ICSI, but do not significantly increase the clinical miscarriage rate. The deleterious impact of type 3 fibroids was remarkable in women with type 3 fibroids with TD or SD >2.0cm.