Intracytoplasmic Sperm Injection Therapy Research Articles

P-524 Bi-allelic CFAP61 variants cause male infertility in humans and mice with severe oligoasthenoteratozoospermia

Abstract Study question Are mutations in cilia and flagella-associated protein 61 (CFAP61) associated with human male infertility? Summary answer Bi-allelic variants ([NM_015585.4: c.1654C>T (p.R552C) and c.2911G>A (p.D971N), c.144-2A>G and c.1666G>A (p.G556R)] in CFAP61 were identified as contributory genetics factor in severe oligoasthenoteratozoospermia (OAT). What is known already Cfap61 knockout mice were infertile due to multiple morphological abnormalities of the sperm flagella (MMAF). However, so far there is no direct evidence that mutations of CFAP61 cause OAT and male infertility. Study design, size, duration Variant screening was performed by whole-exome sequencing (WES) from 325 infertile patients with OAT and 392 fertile individuals. A knockout mouse model was generate to confirm the candidate disease-causing gene, intracytoplasmic sperm injection (ICSI) was used to evaluate the efficiency of clinical treatment. Participants/materials, setting, methods A total 325 OAT-affected patients and 392 men with normal fertility were recruited from China. WES was performed, followed by Sanger sequencing validation. In silico bioinformatics predictions and in vitro functional analyses were performed to evaluate the impacts of candidate disease-causing variants. Hematoxylin and eosin (H&E) staining, electron microscopy, and immunofluorescence assays were performed to evaluate the sperm morphology. Two OAT-affected men with CFAP61 variants were treated by ICSI, and pregnancy outcomes were followed. Main results and the role of chance We identified bi-allelic CFAP61 variants [NM_015585.4: c.1654C>T (p.R552C) and c.2911G>A (p.D971N), c.144-2A>G and c.1666G>A (p.G556R)] in two (0.62%) of the 325 OAT-affected men. In silico bioinformatics analysis predicted that all four variants were deleterious, and in vitro functional analysis confirmed the deleterious effects of the mutants. Notably, H&E staining and electron microscopy analyses of the spermatozoa revealed multiple morphological abnormalities of sperm flagella, the absence of central pair microtubules, and mitochondrial sheath malformation in sperm flagella from man with CFAP61 variants. Further immunofluorescence assays revealed markedly reduced CFAP61 staining in the sperm flagella. In addition, Cfap61-deficient mice showed the OAT phenotype, suggesting that loss of function of CFAP61 was the cause of OAT. Two individuals accepted ICSI therapy using their own ejaculated sperm, and one of them succeeded in fathering a healthy baby. Limitations, reasons for caution Limitations include the lack of in vivo data from the one of patients, and the exact molecular mechanism should be further investigated. Wider implications of the findings Our findings indicate that CFAP61 is essential for spermatogenesis and that bi-allelic CFAP61 variants lead to OAT and male infertility in humans and mice. In addition, our results show that ICSI treatment can be recommended for CFAP61-related OAT. Trial registration number not applicable

Patient with multiple morphological abnormalities of sperm flagella caused by a novel ARMC2 mutation has a favorable pregnancy outcome from intracytoplasmic sperm injection.

To investigate the potential genetic cause in a primary infertility patient with multiple morphological abnormalities of sperm flagella (MMAF). The patient's sperm was observed by light and electron microscopy. Whole-exome sequencing (WES) was carried out to identify candidate genes. Then, the mutation found by WES was verified by Sanger sequencing. The proteins interacting with ARMC2 were revealed by co-immunoprecipitation (co-IP) and mass spectrometry. Intracytoplasmic sperm injection (ICSI) was carried out to achieve successful pregnancy. Typical MMAF phenotype (absent, short, coiled, bent irregular flagella) was shown in the patient's sperm. A novel homozygous mutation in ARMC2 (c.1264C > T) was identified. The proteins interacting with ARMC2 we found were CEP78, PGAM5, RHOA, FXR1, and SKIV2L2. The ICSI therapy was successful, and boy-girl twins were given birth. We found a novel mutation in ARMC2 which led to MMAF and male infertility. This is the first report of ICSI outcome of patient harboring ARMC2 mutation. The interacting proteins indicated that ARMC2 might be involved in multiple processes of spermatogenesis.

Biallelic CFAP61 variants cause male infertility in humans and mice with severe oligoasthenoteratozoospermia

BackgroundThe genetic causes for most male infertility due to severe oligoasthenoteratozoospermia (OAT) remain unclear.ObjectiveTo identify the genetic cause of male infertility characterised by OAT.MethodsVariant screening was performed by whole-exome sequencing...

Ginkgo biloba extract 761 reduces vascular permeability of the ovary and improves the symptom of ovarian hyperstimulation syndrome in a rat model

Aims Ginkgo biloba extract (EGb) has been widely applied in the treatment of cerebrovascular and neurological diseases. However, the effect of EGb761 on ovarian hyperstimulation syndrome (OHSS), a vascular disorder and life-threatening complication of in vitro fertilization and intracytoplasmic sperm injection therapy (IVF/ICSI), has not been evaluated. Materials and methods Forty female Wistar rats aged 22-days old (D22) were divided into eight groups: Control rats received intraperitoneal injection of saline for five consecutive days (D22–D26); OHSS model group received 10 IU equine chorionic gonadotropin (eCG) for four consecutive days (D22–D25) and 30 IU of human chorionic gonadotropin (hCG) on the 5th day (D26); Prophylactic treatment group received three doses of EGb761 (50, 100, and 200 mg/kg/day) 1 h before injection of eCG (hCG) for seven consecutive days; Therapeutic treatment group received three doses of EGb761 (50, 100, and 200 mg/kg/day) 48 h after injection of eCG (hCG) for seven consecutive days. Results All three doses of EGb761 therapeutic medication significantly reduced ovarian mass index of OHSS model rats (p ≤ .01). Furthermore, therapeutic treatment group exhibited improved vascular permeability, decreased estradiol and progesterone levels, lower corpus luteum, and higher follicle numbers compared with the OHSS model. Elevated protein expression of vascular endothelial growth factor (VEGF) and VEGF receptor (VEGFR) in both ovary and kidney of the OHSS model was restrained by EGb761 therapeutic treatment. Conclusions EGb761 therapeutic medication decreases vascular permeability in OHSS rat model by inhibiting VEGF and VEGFR expression, which may contribute to the treatment of OHSS.

Prospective randomized controlled study of a microfluidic chip technology for sperm selection in male infertility patients.

The purpose of this study is to evaluate the impact of a microfluidic approach for spermatozoon selection in male infertility patients undergoing intracytoplasmic sperm injection (ICSI). This research enrolled 128 individuals who had ICSI for male-factor infertility. The patients were separated into two groups according to the method used to pick the spermatozoa: group I (n=64), which used traditional swim-up procedures, and group II (n=64), which used the Fertile Chip for spermatozoon selection during ICSI therapy. Fertilization rates and embryo quality were the major outcomes. The rates of pregnancy, clinical pregnancy and live birth were used as secondary outcomes. As a result, there was no statistically significant difference between the two groups in terms of fertilization rate, total grade 1 and 2 embryos. Implantation rate was significantly higher in the Fertile Chip group than in the control group (50% vs. 31%, p=0.02). The Fertile Chip group had considerably greater pregnancy rates, clinical pregnancy rates (CPR) and live birth rates than the control group (62.5% vs. 45.3%, p=0.038; 59.4% vs. 35.9%, p=0.006 and 46.8% vs. 25%, p=0.009). Fertile Chip had no effect on fertilization rates or embryo quality in male-factor infertility couples. However, the Fertile Chip group had a statistically higher pregnancy rate, CPR and live birth rate.

Ginkgo biloba extract 761 reduces vascular permeability of the ovary and improves the symptom of ovarian hyperstimulation syndrome in a rat model

Aims Ginkgo biloba extract (EGb) has been widely applied in the treatment of cerebrovascular and neurological diseases. However, the effect of EGb761 on ovarian hyperstimulation syndrome (OHSS), a vascular disorder and life-threatening complication of In Vitro Fertilization and Intracytoplasmic Sperm Injection therapy (IVF/ICSI), has not been evaluated. Materials and methods Forty female Wistar rats aged 22-days old (D22) were divided into eight groups: Control rats received intraperitoneal injection of saline for 5 consecutive days (D22-D26); OHSS model group received 10 IU equine chorionic gonadotropin (eCG) for 4 consecutive days (D22-D25) and 30 IU of human chorionic gonadotropin (hCG) on the 5th day (D26); Prophylactic treatment group received three doses of EGb761 (50, 100, and 200 mg/kg/d) one hour before injection of eCG (hCG) for 7 consecutive days; Therapeutic treatment group received three doses of EGb761 (50, 100, and 200 mg/kg/d) 48 h after injection of eCG (hCG) for 7 consecutive days. Results All three doses of EGb761 therapeutic medication significantly reduced ovarian mass index in the OHSS model (p ≤ .01). Further, the therapeutic treatment group exhibited improved vascular permeability, decreased estradiol and progesterone levels, lower corpus luteum, and higher follicle numbers compared with the OHSS model. Elevated protein expression of vascular endothelial growth factor (VEGF) and VEGF receptor (VEGFR) in both ovary and kidney of the OHSS model was restrained by EGb761 therapeutic treatment. Conclusions EGb761 therapeutic medication decreases vascular permeability in OHSS rat model by inhibiting VEGF and VEGFR expression, which may contribute to the treatment of OHSS.

Monitoring of stimulated cycles in assisted reproduction (IVF and ICSI).

Monitoring of in vitro fertilisation (IVF) and intra-cytoplasmic sperm injection (ICSI) is necessary to detect as well as reducethe incidence and severity of ovarian hyperstimulation syndrome (OHSS) whilst achieving the optimal ovarian response needed for assisted reproduction treatment. Traditional monitoring of ovarian hyperstimulation during in vitro fertilisation IVFand ICSItreatment has included transvaginal ultrasonography (TVUS) plus serum estradiol levels. The need for combined monitoring (using TVUS and serum estradiol) during ovarian stimulation in assisted reproduction is controversial. It has been suggested that combined monitoring is time consuming, expensive and inconvenient for women and that simplification of IVF and ICSI therapy by using TVUS only should be considered. OBJECTIVES: To assess the effect of monitoring controlled ovarian hyperstimulation (COH) in IVF and ICSI cycles in subfertile couples with TVUS only versus TVUS plus serum estradiol concentration, with respect to rates of live birth, pregnancy and OHSS. In this update conducted in March 2020, two review authorssearched the Cochrane Gynaecology and Fertility Group's Specialised Register, CENTRAL, MEDLINE, Embase, CINAHL, PsycINFO, the National Research Register, and web-based trial registers. There was no language restriction applied. All references in the identified trials and background papers were checked and authors were contacted to identify relevant published and unpublished data. Only randomised controlled trials that compared monitoring with TVUS only versus TVUS plus serum estradiol concentrations in women undergoing COH for IVF and ICSI treatment were included. Two review authors (IK, AW) independently selected the studies, extracted data and assessed risk of bias. We resolved disagreements by discussion. Outcomes data were pooled and summary statistics were presented when appropriate. The quality of the evidence was rated using the GRADE methods. We did not identify any new eligible studies in this update in 2020. The evidence based on the six trials identified in 2014 remained unchanged. They included 781 women undergoing monitoring of COH with either TVUS alone or a combination of TVUS and serum estradiol concentration during IVF or ICSI treatment. None of the six studies reported our primary outcome of live birth rate. Two studies presented pregnancy rate per initiated cycle and per embryo transfer, respectively. Four studies reported pregnancy rate per woman with pooled data; we are uncertain of the effect ofmonitoring with TVUS only versus combined monitoring on clinical pregnancy rate per woman(odds ratio (OR) 1.10; 95% confidence interval (CI) 0.79 to 1.54; four studies; N = 617; I² = 5%; low quality evidence). This suggests in women with a 36% chance of clinical pregnancy using monitoring with TVUS plus serum estradiol, the clinical pregnancy rate using TVUS only would be between 31% and 46%. We are uncertain of any effectin the mean number of oocytes retrieved per woman (mean difference (MD) 0.32; 95% CI -0.60 to 1.24; five studies; N = 596; I² = 17%; low quality evidence). We are uncertain whethermonitoring with TVUS only versus combined monitoring affectedthe incidence of OHSS (OR 1.03; 95% CI 0.48 to 2.20; six studies; N = 781; I² = 0%; low quality evidence), suggesting that in women with a 4% chance of OHSS using monitoring with TVUS plus serum estradiol, the OHSS rate monitored by TVUS only would be between 2% and 8%. The cycle cancellation rate was similar in both arms of two studies (0/34 versus 1/31, 1/25 versus 1/25; OR 0.57; 95% CI 0.07 to 4.39; N = 115; I² = 0%; low quality evidence). The evidence was low quality for all comparisons. Limitations included imprecision and potential bias due to unclear randomisation methods, allocation concealment and blinding, as well as differences in treatment protocols. Quality assessment was hampered by the lack of methodological descriptions in several studies. This review update found no new randomised trials. Evidence from the six studies previously identified did not suggest that combined monitoring by TVUS and serum estradiol is more efficacious than monitoring by TVUS alone with regard to clinical pregnancy rates and the incidence of OHSS. The number of oocytes retrieved appeared similar for both monitoring protocols. The data suggest that both these monitoring methods are safe and reliable. However, these results should be interpreted with caution because the overall quality of the evidence was low. Results were compromised by imprecision and poor reporting of study methodology.The choice of one or the other method may depend upon the convenience of its use, and the associated costs.An economic evaluation of the costs involved with the two methods and the views of the women undergoing cycle monitoring would be welcome.

Effects of different polyvinylpyrrolidone concentrations on intracytoplasmic sperm injection.

To explore whether different polyvinylpyrrolidone (PVP) concentrations affect the results of intracytoplasmic sperm injection (ICSI), a prospective study was conducted for 194 couples undergoing 210 ICSI therapy cycles. These cycles were divided into three groups (10, 7 and 5% groups) using the corresponding concentration of PVP for sperm immobilization. The main outcome measures were analyzed. Results indicated that, with a decrease in PVP concentrations, all of the main outcome measures increased. In particular, the high-quality cleavage embryo rate in the 7% group was significantly lower than in the 5% group (P < 0.01), and the cleavage, high-quality cleavage embryo, and high-quality blastocyst rates in the 5% group were significantly higher than those in the 10% group (all P < 0.001). For high-/intermediate-quality semen, all of the main outcome measures were significantly increased with 5% PVP. For the poor-quality semen, only the high-quality cleavage embryo and high-quality blastocyst rates were significantly higher in the 5% group. Therefore, lowering PVP concentrations greatly promoted the development of embryos in ICSI cycles, with an optimal concentration of 5% for ICSI.

Paternal Smoking in Relation to Sperm Quality and intracytoplasmic Sperm Injection Outcomes

Objectives: The present study focused on tobacco smoke and its effect on semen parameters, sperm DNA quality (compaction and fragmentation) and clinical outcomes after intracytoplasmic sperm injection (ICSI) therapy Materials and Methods: The semen samples were divided according to smoking status into the following 2 groups, 98 heavy-smokers (G1) and 43 non-smokers (G2). Semen was prepared and purified using the PureSperm gradients according to the WHO guidelines 2010. Protamine deficiency (CMA3 positivity) was assessed by chromomycin CMA3 staining and sperm DNA fragmentation (sDF) by TUNEL assay. Results: The mean concentration and the total motility were significantly higher in G2 in comparison to G1 (P=0.014, and P=0.026 respectively) and the results were similar for the mean percent of the progressive motility and normal morphology (P=0.0001). CMA3+ and sDF in G2 were significantly lower in comparison to G1 (20.35 ± 13.34% vs. 33.30 ± 22.33%, P=0.001; 14.23 ± 13.07% vs. 26.68 ± 19.77%, P=0.0001). Meanwhile, there were no significant differences in the ICSI outcomes, except for the pregnancy rate, which was significantly higher in G2 than in G1 (0.60 ± 0.49% vs. 0.38 ± 0.48%; P=0.013). In G1, CMA3+ correlated negatively with sperm concentration (r=-0.233, P=0.021) but positively with sDF (r=0.484, P=0.0001). In G2, sDF correlated negatively with progressive motility and morphologically normal spermatozoa (r=-0.304, p=0.047; r=-0.361, P=0.017 respectively). Conclusions: The findings of this study revealed that tobacco smoking altered sperm parameters and later affected the pregnancy results in ICSI therapy. CMA3 and TUNEL tests are therefore useful as a supplementary test before any ART treatment to ensure a good prognosis.

The need to improve patient care through discriminate use of intracytoplasmic sperm injection (ICSI) and improved understanding of spermatozoa, oocyte and embryo biology

D. T. Carrell, A. Nyboe Andersen and D. J. Lamb Departments of Surgery (Urology), Obstetrics and Gynecology, and Human Genetics, University of Utah School of Medicine, Salt Lake City, UT, USA, The Fertility Clinic, Copenhagen University Hospital, Rigshospitalet, Copenhagen, Denmark, and The Center for Reproductive Medicine, Scott Department of Urology and the Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX, USA

Monitoring of stimulated cycles in assisted reproduction (IVF and ICSI).

Traditional monitoring of ovarian hyperstimulation during in vitro fertilisation (IVF) and intra-cytoplasmic sperm injection (ICSI) treatment has included transvaginal ultrasonography (TVUS) plus serum estradiol levels to ensure safe practice by reducing the incidence and severity of ovarian hyperstimulation syndrome (OHSS) whilst achieving the good ovarian response needed for assisted reproduction treatment. The need for combined monitoring (using TVUS and serum estradiol) during ovarian stimulation in assisted reproduction is controversial. It has been suggested that combined monitoring is time consuming, expensive and inconvenient for women and that simplification of IVF and ICSI therapy by using TVUS only should be considered. To assess the effect of monitoring controlled ovarian hyperstimulation (COH) in IVF and ICSI cycles in subfertile couples with TVUS only versus TVUS plus serum estradiol concentration, with respect to rates of live birth, pregnancy and OHSS. We searched the Menstrual Disorders and Subfertility Group Specialised Register of controlled trials, the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, EMBASE, CINAHL, PsycINFO, the National Research Register, and web-based trial registers such as Current Controlled Trials. The last search was conducted in May 2014. There was no language restriction applied. All references in the identified trials and background papers were checked and authors were contacted to identify relevant published and unpublished data. Only randomised controlled trials that compared monitoring with TVUS only versus TVUS plus serum estradiol concentrations in women undergoing COH for IVF and ICSI treatment were included. Three review authors independently selected the studies, extracted data and assessed risk of bias. They resolved disagreements by discussion with the rest of the authors. Outcomes data were pooled and summary statistics were presented when appropriate. The quality of the evidence was rated using the GRADE methods. With this update, four new studies were identified resulting in a total of six trials including 781 women undergoing monitoring of COH with either TVUS alone or a combination of TVUS and serum estradiol concentration during IVF or ICSI treatment.None of the six studies reported our primary outcome of live birth rate. Pooled data showed no evidence of a difference in clinical pregnancy rate per woman between monitoring with TVUS only and combined monitoring (odds ratio (OR) 1.10; 95% confidence interval (CI) 0.79 to 1.54; four studies; N = 617; I² = 5%; low quality evidence). This suggests that compared with women with a 34% chance of clinical pregnancy using monitoring with TVUS plus serum estradiol, the clinical pregnancy rate in women using TVUS only was between 29% and 44%.There was no evidence of a difference between the groups in the reported cases of OHSS (OR 1.03; 95% CI 0.48 to 2.20; six studies; N = 781; I² = 0%; low quality evidence), suggesting that compared with women with a 4% chance of OHSS using monitoring with TVUS plus serum estradiol, the OHSS rate in women monitored by TVUSS only was between 2% and 8%.There was no evidence of a difference between the groups in the mean number of oocytes retrieved pre woman (mean difference (MD) 0.32; 95% CI -0.60 to 1.24; five studies; N = 596; I² = 17%; low quality evidence).The evidence was low quality for all comparisons. Limitations included imprecision and potential bias due to unclear randomisation methods, allocation concealment and blinding, as well as differences in treatment protocols. Quality assessment was hampered by the lack of methodological descriptions in several studies. This review update found no evidence from randomised trials to suggest that combined monitoring by TVUS and serum estradiol is more efficacious than monitoring by TVUS alone with regard to clinical pregnancy rates and the incidence of OHSS. The number of oocytes retrieved appeared similar for both monitoring protocols. The data suggest that both these monitoring methods are safe and reliable. However, these results should be interpreted with caution because the overall quality of the evidence was low. Results were compromised by imprecision and poor reporting of study methodology. A combined monitoring protocol including both TVUS and serum estradiol may need to be retained as precautionary good clinical practice and as a confirmatory test in a subset of women to identify those at high risk of OHSS. An economic evaluation of the costs involved with the two methods and the views of the women undergoing cycle monitoring would be welcome.

Study on effects of sperm morphology to some parameter of ICSI

Objective To explore the influence of sperm morphology to some consequence parameter of intracytoplasmic sperm injection (ICSI), by sperm morphology analysis and statistical analysis to infertility patients in ICSI therapy. Methods We accounted the rate of morphologically normal sperm by Kruger' s strict criteria , and analyzed the normal sperm morphology rate and the rates of fertilization, cleavage, embryo quality, embryo transfer and clinical pregnancy by correlation analysis,rank-sum test and chi square test. Reaults The normal sperm morphology rate did not show any correlation with the rates of fertilization, cleavage, embryo quality and embryo transfer;, no statistic differences were found in the rates of fertilization, cleavage, embryo quality, embryo transfer and clinical pregnancy between the normal sperm morphology rate 0.05). Conclusion Sperm morphology does not influence some consequence parameter of ICSI, such as the rates of fertilization,cleavage, embryo quality, embryo transfer and clinical pregnancy. Key words: Sperm morphology; Normal sperm morphology rate; ICSI; Fertilization rate; Clinical pregnancy rate

A sperm viability test using SYBR-14/propidium iodide flow cytometry as a tool for rapid screening of primary ciliary dyskinesia patients and for choosing sperm sources for intracytoplasmic sperm injection

Spermatozoa viability tests based on dual-color flow cytometry after staining with Sybr-14/propidium iodide were performed on 44 men with complete asthenospermia for primary ciliary dyskinesia (PCD) screening, and seven were identified with PCD by electron microscopy of ultrastructural ciliary defects. Six PCD patients underwent eight intracytoplasmic sperm injection therapy cycles using ejaculated sperm or testicular sperm, obtaining a mean fertilization rate of 46.6%, with three healthy babies born and one in utero at the time of writing.

The role of paternal chromosomes and sperm morphology on the outcome of intracytoplasmic sperm injection

The study investigated the possible relationship between the X/Y chromosomes bearing spermatozoa and the outcome of intracytoplasmic sperm injection (ICSI) therapy and morphological appearance/shape (Tygerberg criteria) of the sperm cell injected into the oocyte during ICSI therapy and fertilisation. Thirty-nine patients were recruited for the study from an assisted reproductive programme. Semen samples were prepared by using gradient centrifugation techniques. Prior to injection sperm images were captured using high-quality video graphic equipment. Sperm selection was based on the concept of 'best-looking' spermatozoa i.e. spermatozoa lacking gross and obvious malformations such as broken necks, cytoplasmic droplets, amorphous or elongated heads. Photomicrographs of each sperm cell were produced from video footage. The photographical material was used to determine the basic shape and the actual length-to-width ratio of the injected sperm heads. Embryo biopsies and fluorescent in situ hybridisation (FISH) was performed on 12 randomly selected couples from a set of 39. Embryos were evaluated on day 3 for development and embryo transfer. Embryo biopsies and FISH analyses were performed on those embryos that showed no developmental potential following injection. It was found that 70% of the embryos that showed no developmental potential were Y chromosome-bearing embryos. The sperm selection process for ICSI based on the approach of choosing the 'best-looking' spermatozoon in the ejaculate seems to provide cells that can be classified as normal based on the length-to-width ratio set by the World Health Organization for normal cells.

Centrosomal function assessment in human sperm using heterologous ICSI with rabbit eggs: a new male factor infertility assay.

Sperm centrosomal function was assessed by immunocytochemical analysis after the injection of human sperm into mature rabbit eggs. Three hours after intracytoplasmic sperm injection (ICSI), an astral microtubule array from the base of the human sperm was observed in the rabbit eggs. This sperm aster expanded in the egg cytoplasm, concomitant with pronuclear formation, and a dense microtubule array was organized at the time of pronuclear centration. Using fertile donor sperm, the sperm aster formation rate at 3 hr after ICSI was 35.0 +/- 1.5%. Using sperm from infertile patients, the average aster formation rate was lower (25.4 +/- 14.8%, P<0.05). Among infertile cases, there was no correlation between sperm aster formation rates and conventional parameters of semen analysis. However, the sperm aster formation rate correlated with the embryonic cleavage rate following human in vitro fertilization (IVF). These data suggest that this assay reflects sperm function during embryonic development after sperm entry and that reproductive success during the first cell cycle requires a functional sperm centrosome. Furthermore, sperm centrosomal function cannot be predicted from conventional parameters of semen analysis. We propose that insufficient centrosomal function could be the cause of certain cases of idiopathic infertility. These assays may lead to the discovery of new types of infertility, which have previously been treated as "unexplained infertility," and may also lead to the treatment of infertility incurable even by ICSI. Consequently, an accurate and relevant assay to help assure couples of the success of fertilization is warranted, perhaps prior to ICSI therapy.

Relationship between ovarian stimulation regimen and interleukin level in pre-ovulatory follicular fluid and their effect on ICSI outcome.

The aims of this study were (i) to identify the presence and concentration of interleukin-6 (IL-6), IL-8 and granulocyte-macrophage colony-stimulating factor (GM-CSF) in pre-ovulatory follicular fluid (FF) in patients undergoing controlled ovarian hyperstimulation regimen for intracytoplasmic sperm injection (ICSI) with recombinant human follicle stimulating hormone (rFSH) (G.1), hMG (G.2) or rFSH/ hMG (G.3) treatment and (ii) to determine whether controlled ovarian hyperstimulation stimulation regimen could affect the concentration of these investigated interleukins and as a consequences the ICSI outcome. (iii) Moreover, it was intended to clarify the association between these investigated parameters and the peripheral blood level of estradiol 17-beta, progesterone, leuteinizing hormone (LH) at the day of oocyte retrieval and (iv) finally to find out whether these interleukin concentrations in FF could be considered as an additional parameter for predicting fertilization and pregnancy outcome in ICSI patients. Interleukin-6, IL-8 and GM-CSF concentrations were determined in the FF of 85 patients. The levels of these interleukins were measured by the enzyme-linked immunosorbent assay (ELISA) technique, using commercially available kits. Interleukin-6 (6.5 +/- 4.6 pg/mL), IL-8 (222.9 +/- 163.4 pg/mL) and GM-CSF (1.6 +/- 2.7 pg/mL) were present in pre-ovulatory FF in ICSI patients. No significant difference in the mean concentration of IL-6, IL-8 and GM-CSF was found between the groups. Moreover, there was no correlation between these interleukin levels in FF and steroid hormone concentration in the peripheral blood. Interleukin-6, IL-8 and GM-CSF were found in FF of patients undergoing ovarian hyperstimulation for ICSI treatment. The stimulation protocols showed no influence on these interleukin levels in FF. Moreover, no correlation was found between these investigated interleukins and steroid hormone concentrations in peripheral blood or ICSI outcome. Therefore, the determination of interleukin level in FF of patients undergoing ovarian hyperstimulation for ICSI therapy could not be recommended as a prognostic factor for ICSI outcome.

Relationship between cytokine concentration in serum, pre‐ovulatory follicular fluid and icsi outcome

This study was undertaken to determine the concentrations of fibroblast growth factor (FGF), stem cell factor (SCF) and soluble intracellular adhesion molecule (sICAM) in serum and follicular fluid (FF) of patients who became pregnant (G.I) and those who did not (G.II) after controlled ovarian hyperstimulation (COH) for intracytoplasmic sperm injection (ICSI) therapy and to find out whether any relationship could exist between cytokine concentration in serum, FF on the day of oocytes retrieval and ICSI outcome.Serum and FF was collected from 75 consecutive patients undergoing COH for ICSI therapy after pituitary down regulation with GnRHa. Three ampules (225 IU) of (hMG) were administered daily from day 3–7 of the cycle. 10.000 IU HCG was given i.m. Follicular aspiration was performed 35–36 hr after HCG injection. These cytokines were measured with ELISA assay kits. The cytokine levels in the serum of (G.I) were 8.5 + 10.5 pg/mL, 235.8 + 81.10 ng/mL and 597.7 + 139.9 pg/mL and the corresponding concentrations of (G.II) were (6.4 + 3.6, 230.6 + 66.5 and 569.6 + 91.4, respectively). There was no significant difference between the cytokines concentration in serum and FF in the groups. However, in both groups, the concentration of FGF and SCF in FF was significantly higher (P = 0.0001 and P = 0.05) than the value observed in serum. Whereas, the sICAM concentration was significantly higher of serum in comparison to FF value. The two groups did not differ significantly for the total number of gonadotropin ampoules administered, oestradiol concentration on the day of HCG injection, number of retrieved oocytes, and fertilization rate. In conclusion: Group I (the 25 women who did become pregnant) and group II (the 50 women who did not become pregnant) did not differ significantly for any of these cytokines concentrations. Besides, the ICSI outcome was not related to serum or follicular fluid concentrations of FGF, sICAM and SCF concentrations.

Low-level sex chromosome mosaicism in female partners of couples undergoing ICSI therapy does not significantly affect treatment outcome.

There is an increased rate of chromosomal anomalies, in particular low-level sex chromosome mosaicism, in the female partners of couples undergoing intracytoplasmic sperm injection (ICSI). Among 811 consecutive couples presenting for pre-ICSI chromosome analysis, chromosomal abnormalities were detected in 54 individuals, of which 26 were low-level sex chromosome mosaicism in the females. Attention was focused on the treatment course and outcome of ICSI in 20 couples with low-level sex chromosome mosaicism in the females actually embarking on ICSI treatment (group I, n = 38 ICSI treatment cycles). Applying a case-control design, each of the 20 couples was matched according to female age and source of spermatozoa to couples without a chromosomal abnormality in either of the partners (group II, n = 38 ICSI treatment cycles). No significant differences were found between the groups in ovarian response, fertilization rate and number of embryos transferred. Pregnancy rates, as well as implantation and abortion rates did not differ significantly between the groups. The data suggest that low-level sex chromosome mosaicism in females has no major effect on the course and outcome of ICSI.

Association between interleukin concentration in follicular fluid and intracytoplasmic sperm injection (ICSI) outcome.

The aim of this study was to determine the presence and concentration of interleukin IL-6, IL-8, and granulocyte-macrophage-colony-stimulating factor (GM-CSF) in pre-ovulatory ovarian follicular fluid (FF) of patients undergoing controlled ovarian hyperstimulation for intracytoplasmic sperm injection (ICSI) therapy on one hand, and to find out whether these cytotokine concentrations could be used as a predictive parameter for ICSI outcome. The levels of IL-6, IL-8, and GM-CSF were measured from women that underwent ICSI therapy and the results were compared between the patients who became pregnant after IC

Clinical importance of zona pellucida-induced acrosome reaction and its predictive value for IVF.

The study aimed to establish zona pellucida induced acrosome reaction response (ZIAR) among 35 couples with normal and G-pattern sperm morphology and repeated poor fertilization results during assisted reproduction treatment. ZIAR tests were performed using 0.25 zona pellucida/microliter co-incubated with spermatozoa for 60 min. Acrosome reactions were measured with FITC-PSA staining, and expressed as the difference between stimulated and unstimulated (spontaneous) sperm populations. Results were compared with IVF rates of metaphase II oocytes. Interactive dot diagrams divided the patients into two groups, i.e. ZIAR <15% and ZIAR > 15%, with mean fertilization rates of 49 and 79% respectively. The sensitivity and specificity for ZIAR results versus fertilization were 93 and 100% respectively. The area under the curve was 99% and the 95% confidence interval did not include 0.5 which implies that the ZIAR test is able to predict fertilization failure among IVF patients. In conclusion, the ZIAR test has diagnostic potential since it can assist the clinician to identify couples that will benefit from intracytoplasmic sperm injection therapy.