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ICSI
Selected AbstractsAssisted reproductive technologies and birth defectsCONGENITAL ANOMALIES, Issue 2 2005Kohei Shiota ABSTRACT In vitro fertilization (IVF) and other assisted reproductive technologies (ART) are effective treatments for infertility and are widely provided at infertility clinics. Although IVF and related ART procedures are generally considered safe, some studies have suggested an excess occurrence of major malformations, low birth-weight and other perinatal complications in babies conceived by ART. Further, it was recently reported that IVF and intracytoplasmic sperm injection (ICSI) are associated with imprinting disorders in the offspring such as Beckwith-Wiedemann syndrome and Angelman syndrome. Here we review the human and animal studies investigating the potential risks of ART, and discuss the need for further investigation. [source] Fertility treatment in male cancer survivorsINTERNATIONAL JOURNAL OF ANDROLOGY, Issue 4 2007Kirsten Louise Tryde Schmidt Summary The present study reviews the use of assisted reproductive technology in male cancer survivors and their partners. As antineoplastic treatment with chemotherapy or radiation therapy, has the potential of inducing impairment of spermatogenesis through damage of the germinal epithelium, many male cancer survivors experience difficulties in impregnating their partners after treatment. The impairment can be temporary or permanent. While many cancer survivors regain spermatogenesis months to years after treatment, some become infertile with a-, oligo- or azoospermia. An option to secure the fertility potential of young cancer patients is to cryopreserve semen before cancer treatment for later use. A desired pregnancy may be obtained in couples where the husband has a history of cancer, using assisted reproductive technology with either fresh or cryopreserved/thawed semen. Successful outcomes have been obtained with intrauterine insemination (IUI) as well as in vitro fertilization (IVF) with or without the use of intracytoplasmic sperm injection (ICSI). In conclusion, male cancer survivors and their partners who have failed to obtain a pregnancy naturally within a reasonable time frame after end of treatment should be referred to a fertility clinic. [source] Intracytoplasmic sperm injection as a complement to gonadotrophin treatment in infertile men with hypogonadotrophic hypogonadismINTERNATIONAL JOURNAL OF ANDROLOGY, Issue 4 2005BRANKO ZORN Summary In this study we sought to determine whether intracytoplasmic sperm injection (ICSI) could improve the efficacy of treatment with gonadotrophins in gonadotrophin-deficient men in terms of pregnancy. A series of six adult men (aged 26,47 years) with hypogonadotrophic hypogonadism (HH) is reported: four men with prepubertal isolated idiopathic HH (IIHH) and two adult-onset HH, as part of hypopituitarism secondary to surgical treatment of a pituitary tumour. All were azoospermic. To restore spermatogenesis, all received hormonal treatment with intramuscular human menopausal gonadotrophins (HMG) and human chorionic gonadotrophin (HCG) for 2 to 23 months. High basal serum inhibin B was predictive of rapid and complete recovery of spermatogenesis. In the two adult-onset HH, a natural pregnancy was achieved within 3 months. The four men with IIHH underwent ICSI because of poor sperm quality. ICSI using fresh or frozen-thawed ejaculated spermatozoa was performed after 6,23 months of gonadotrophin treatment. ICSI provided good clinical results in terms of fertilization and embryo quality, and resulted in three pregnancies that ended in three term deliveries. In men with oligozoospermia related to prepubertal IIHH, ICSI shortens the hormonal treatment and enhances the chances of pregnancy. [source] Comparison between computerized slow-stage and static liquid nitrogen vapour freezing methods with respect to the deleterious effect on chromatin and morphology of spermatozoa from fertile and subfertile menINTERNATIONAL JOURNAL OF ANDROLOGY, Issue 2 2001M. E. Hammadeh The purpose of this study was to determine the negative effects (cryodamage) on human spermatozoa after freeze-thawing and to determine whether freeze-thawing of spermatozoa with a programmed slow freezer is better than freezing with liquid nitrogen vapour (rapid freezing) with regard to alterations in sperm chromatin and morphology in semen from fertile (donor) and subfertile, IVF/ICSI, patients. Ninety-five semen samples were obtained either from patients attending our IVF unit for treatment (n=34) or from donors (n=25) with proven fertility and normal sperm quality according to WHO guidelines. Each semen sample was divided into two parts after liquefaction and addition of the cryoprotectant. The first part was frozen using a programmed biological freezer and the second part was frozen by means of liquid nitrogen vapour. Smears were made before the freezing and after the thawing procedure to assess morphology (strict criteria) and chromatin condensation (Acridine Orange test). The mean percentage of chromatin condensed spermatozoa in the samples from donors (control group) was 92.4 ± 8.4% before freezing and this decreased significantly (p < 0.0001) to 88.7 ± 11.2% after freeze-thawing with the computerized slow-stage freezer and to 87.2 ± 12.3% after using static liquid nitrogen vapour (p < 0.001). The corresponding values for semen obtained from patients was 78.9 ± 10.3% before freezing which decreased to 70.7 ± 10.8 and 68.5 ± 14.8%, respectively (p < 0.001). On the other hand, the mean percentage of normal sperm morphology in the control group decreased from 26.3 ± 7.5% before freezing to 22.1 ± 6.4% (p < 0.0001) after thawing with the computerized slow-stage freezer and to 22.2 ± 6.6% (p < 0.0001) after the use of static liquid nitrogen vapour. In the patient group, the mean percentage of normal morphology decreased from 11.7 ± 6.1% after freezing with the biological freezer to 9.3 ± 5.6% and to 8.0 ± 4.9% after freezing with static liquid nitrogen vapour. This study demonstrates that chromatin packaging and morphology of human spermatozoa decrease significantly after the freeze-thawing procedure, not only after the use of static liquid nitrogen vapour but also after the use of a computerized slow-stage freezer. However, the chromatin of semen samples with normal semen parameters (donor sperm) withstand the freeze-thaw injury better than those with low quality semen samples. Therefore, the computerized slow stage freezer could be recommended for freezing of human spermatozoa, especially for subnormal semen samples, for example, ICSI and ICSI/TESE candidates and from patients with testicular tumours or Hodgkin's disease, in order to avoid further damage to the sperm chromatin structure. [source] Bayesian statistics in medical research: an intuitive alternative to conventional data analysisJOURNAL OF EVALUATION IN CLINICAL PRACTICE, Issue 2 2000AStat, Lyle C. Gurrin BSc (Hons) Summary Statistical analysis of both experimental and observational data is central to medical research. Unfortunately, the process of conventional statistical analysis is poorly understood by many medical scientists. This is due, in part, to the counter-intuitive nature of the basic tools of traditional (frequency-based) statistical inference. For example, the proper definition of a conventional 95% confidence interval is quite confusing. It is based upon the imaginary results of a series of hypothetical repetitions of the data generation process and subsequent analysis. Not surprisingly, this formal definition is often ignored and a 95% confidence interval is widely taken to represent a range of values that is associated with a 95% probability of containing the true value of the parameter being estimated. Working within the traditional framework of frequency-based statistics, this interpretation is fundamentally incorrect. It is perfectly valid, however, if one works within the framework of Bayesian statistics and assumes a ,prior distribution' that is uniform on the scale of the main outcome variable. This reflects a limited equivalence between conventional and Bayesian statistics that can be used to facilitate a simple Bayesian interpretation based on the results of a standard analysis. Such inferences provide direct and understandable answers to many important types of question in medical research. For example, they can be used to assist decision making based upon studies with unavoidably low statistical power, where non-significant results are all too often, and wrongly, interpreted as implying ,no effect'. They can also be used to overcome the confusion that can result when statistically significant effects are too small to be clinically relevant. This paper describes the theoretical basis of the Bayesian-based approach and illustrates its application with a practical example that investigates the prevalence of major cardiac defects in a cohort of children born using the assisted reproduction technique known as ICSI (intracytoplasmic sperm injection). [source] Stimulation day-six serum estradiol: A predictive indicator for the probability of embryo cryopreservation in IVF/ICSI cyclesJOURNAL OF OBSTETRICS AND GYNAECOLOGY RESEARCH (ELECTRONIC), Issue 2 2009Hassan A. El Maghraby Abstract Objective:, To evaluate the predictive value of stimulation day six serum estradiol (E2) for the probability of embryo cryopreservation after fresh embryo transfer in intracytoplasmic sperm injection (ICSI) cycles. Subjects and Methods:, The study included 282 ICSI cycles for different causes of infertility, provided that the age of the female partner was <40 years and her basal follicle stimulating hormone <10 IU/L. Setting:, Alexandria IVF/ICSI center. Main Outcome Measures:, Primary outcome measures are stimulation day-six serum E2, and rate of embryo cryopreservation, after transfer of three good-quality embryos. Secondary outcome measures are pregnancy rate per fresh embryo transfer, and other intermediate variables of the ICSI cycle. Results:, Patients were stratified into three groups according to day-six serum E2 levels: Group I with values <400 pg/mL; Group II, between 400 and 900; and Group III with values >900. The mean number of oocytes retrieved was 6.3, 8.9, and 12.4; the mean number of obtained embryos was 3.3, 4.8, and 6.7; and pregnancy rates were 18.1, 36.2, and 44.7% in the three groups, respectively. Rate of embryo cryopreservation, after transfer of three good-quality embryos was 70.7% in Group III, and 26.5% in Group I. (P = 0.01). The negative predictive value of day-six E2 < 400 pg/mL for freezing was 83% while day-six serum E2 > 900 pg/mL has a sensitivity of 55%, specificity of 72% and positive predictive value of 50% for embryo freezing. Conclusion:, Higher stimulation day-six estradiol was associated with a higher probability of cryopreservation, and a higher pregnancy rate. [source] Relationship between fertilizing ability of ejaculated human spermatozoa and its chromatin heterogeneityJOURNAL OF OBSTETRICS AND GYNAECOLOGY RESEARCH (ELECTRONIC), Issue 2 2002Shinako Hashimoto Objective: The aim of this study was to examine the relationship between the fertilizing ability of ejaculated human sperm and its chromatin heterogeneity. Methods: We used the D-AO staining method (acridine orange epifluorescence accompanied by diamide, a thiol oxidizing agent) to analyze the sperm chromatin structure of infertile patients with IVF-ET treatment. SDS-PAGE was performed to analyze the sperm nuclear proteins collected from patients with immature sperm (stained red with D-AO staining) and proven fertile men. Results (1) It was suggested that D-AO staining allowed the immature sperm to be divided into two groups. One was immature sperm, which had disturbance of S-S formation in the epididymides, and the other was that which had an abnormal exchange process of nuclear proteins in the testes. (2) The stainability after D-AO staining showed no correlation with the findings of semen analysis. (3) The fertilization rate of IVF-ET was significantly correlated to the percentage of green sperm staining with D-AO staining. (4) In the pregnant group after IVF-ET, it was noticed that sperm of the green type with D-AO staining was increased in comparison with the non-pregnancy group. (5) The fertilization rate in the group of the sperm stained red with D-AO staining was increased to 73.5% by ICSI. (6) Definite differences were noticed between the protein components of the patients with immature sperm and those of the proven fertile men by analyzing with SDS-PAGE. Conclusion D-AO staining was an efficient method for evaluating the fertilizing ability of human ejaculated sperm, and to determine an appropriate ART tool such as ICSI. [source] Middle Eastern Masculinities in the Age of New Reproductive Technologies: Male Infertility and Stigma in Egypt and LebanonMEDICAL ANTHROPOLOGY QUARTERLY, Issue 2 2004MARCIA C. INHORNArticle first published online: 8 JAN 200 Worldwide, male infertility contributes to more than half of all cases of childlessness; yet, it is a reproductive health problem that is poorly studied and understood. This article examines the problem of male infertility in two Middle Eastern locales, Cairo, Egypt, and Beirut, Lebanon, where men may be at increased risk of male infertility because of environmental and behavioral factors. It is argued that male infertility may be particularly problematic for Middle Eastern men in their pronatalist societies; there, both virility and fertility are typically tied to manhood. Thus, male infertility is a potentially emasculating condition, surrounded by secrecy and stigma. Furthermore, the new reproductive technology called intracytoplasmic sperm injection (ICSI), designed specifically to overcome male infertility, may paradoxically create additional layers of stigma and secrecy, due to the complex moral and marital dilemmas associated with Islamic restrictions on third-party donation of gametes. [male infertility, masculinity, new reproductive technologies, stigma, Egypt, Lebanon] [source] Activation regimens for full-term development of rabbit oocytes injected with round spermatidsMOLECULAR REPRODUCTION & DEVELOPMENT, Issue 6 2009Masumi Hirabayashi The present study was designed to investigate the effect of activation regimens on full-term development of rabbit oocytes after round spermatid injection (ROSI). In the first series, rabbit oocytes were treated with 5 µM ionomycin before ROSI, after ROSI, or before and after ROSI. In addition, non-treated oocytes were subjected to intracytoplasmic sperm injection (ICSI) using ejaculated spermatozoa. Cleavage rate of ROSI oocytes activated before and after ROSI (55%) was comparable with that of ICSI oocytes (60%), and significantly higher than those of ROSI oocytes activated either before or after ROSI (29,39%; P,<,0.05). No offspring were produced by transfer of the cleaving ROSI oocytes, while 8% of the cleaving ICSI oocytes transferred gave birth to offspring. In the second series, oocytes were exposed to 5, 10, or 20 µM ionomycin, followed by ROSI, 5 µM ionomycin treatment, and incubation with 5 µg/ml cycloheximide (CHX),+,2 mM 6-dimethylaminopurine (DMAP). Significantly higher cleavage rates were derived from oocytes activated with 10 and 20 µM ionomycin before ROSI (91% and 82%, respectively; P,<,0.05) compared to those activated with 5 µM ionomycin before ROSI (53%). Live offspring were obtained when the cleaving ROSI oocytes with the initial ionomycin treatment at 5 and 10 µM were transferred (offspring rate 2% and 4%, respectively). These activation regimens, however, were not valid for the ROSI using cryopreserved round spermatids. In conclusion, rabbit ROSI oocytes were capable of developing into full-term when the oocytes were activated with a combined treatment of ionomycin and CHX/DMAP. Mol. Reprod. Dev. 76: 573,579, 2009. © 2008 Wiley-Liss, Inc. [source] Functional assessment of centrosomes of spermatozoa and spermatids microinjected into rabbit oocytes,MOLECULAR REPRODUCTION & DEVELOPMENT, Issue 3 2009Masahito Tachibana Abstract Although intracytoplasmic sperm injection (ICSI) is a widely used assisted reproductive technique, the fertilization rates and pregnancy rates of immature spermatids especially in round spermatid injection (ROSI) remain very low. During mammalian fertilization, the sperm typically introduces its own centrosome which then acts as a microtubule organizing center (MTOC) and is essential for the male and female genome union. In order to evaluate the function of immature germ cell centrosomes, we used the rabbit gamete model because rabbit fertilization follows paternal pattern of centrosome inheritance. First, rabbit spermatids and spermatozoa were injected into oocytes using a piezo-micromanipulator. Next, the centrosomal function to form a sperm aster was determined. Furthermore, two functional centrosome proteins (,-tubulin and centrin) of the rabbit spermatogenic cells were examined. Our results show that the oocyte activation rates by spermatozoa, elongated spermatids, and round spermatids were 86% (30/35), 30% (11/36), and 5% (1/22), respectively. Sperm aster formation rates after spermatozoa, elongated spermatids, and round spermatids injections were 47% (14/30), 27% (3/11), and 0% (0/1), respectively. The aster formation rate of the injected elongating/elongated spermatids was significantly lower than that of the mature spermatozoa (P,=,0.0242). Moreover, sperm asters were not observed in round spermatid injection even after artificial activation. These data suggest that poor centrosomal function, as measured by diminished aster formation rates, is related to the poor fertilization rates when immature spermatogenic cells are injected. Mol. Reprod. Dev. 76: 270,277, 2009. © 2008 Wiley-Liss, Inc. [source] Effect of maturation stage at cryopreservation on post-thaw cytoskeleton quality and fertilizability of equine oocytesMOLECULAR REPRODUCTION & DEVELOPMENT, Issue 5 2006T. Tharasanit Abstract Oocyte cryopreservation is a potentially valuable technique for salvaging the germ-line when a valuable mare dies, but facilities for in vitro embryo production or oocyte transfer are not immediately available. This study examined the influence of maturation stage and freezing technique on the cryopreservability of equine oocytes. Cumulus oocyte complexes were frozen at the immature stage (GV) or after maturation in vitro for 30 hr (MII), using either conventional slow freezing (CF) or open pulled straw vitrification (OPS); cryoprotectant-exposed and untreated nonfrozen oocytes served as controls. After thawing, GV oocytes were matured in vitro, and MII oocytes were incubated for 0 or 6 hr, before staining to examine meiotic spindle quality by confocal microscopy. To assess fertilizability, CF MII oocytes were subjected to intracytoplasmic sperm injection (ICSI) and cultured in vitro. At 12, 24, and 48 hr after ICSI, injected oocytes were fixed to examine their progression through fertilization. Both maturation stage and freezing technique affected oocyte survival. The meiosis resumption rate was higher for OPS than CF for GV oocytes (28% vs. 1.2%; P,<,0.05), but still much lower than for controls (66%). Cryopreserving oocytes at either stage induced meiotic spindle disruption (37%,67% normal spindles vs. 99% in controls; P,<,0.05). Among frozen oocytes, however, spindle quality was best for oocytes frozen by CF at the MII stage and incubated for 6 hr post-thaw (67% normal); since this combination of cryopreservation/IVM yielded the highest proportion of oocytes reaching MII with a normal spindle (35% compared to <20% for other groups), it was used when examining the effects of cryopreservation on fertilizability. In this respect, the rate of normal fertilization for CF MII oocytes after ICSI was much lower than for controls (total oocyte activation rate, 26% vs. 56%; cleavage rate at 48 hr, 8% vs. 42%: P,<,0.05). Thus, although IVM followed by CF yields a respectable percentage of normal-looking MII oocytes (35%), their ability to support fertilization is severely compromised. Mol. Reprod. Dev. © 2006 Wiley-Liss, Inc. [source] Influence of oocyte collection technique on initial chromatin configuration, meiotic competence, and male pronucleus formation after intracytoplasmic sperm injection (ICSI) of equine oocytes,MOLECULAR REPRODUCTION & DEVELOPMENT, Issue 1 2001Maria Elena Dell'Aquila Abstract There is a great variability in the success of horse oocyte maturation and fertilization among laboratories. This study was conducted to determine if the meiotic and developmental competence of horse oocytes could be dependent on the method of oocyte collection, i.e., aspiration of follicular fluid with a vacuum apparatus, or opening follicles and scraping the granulosa layer. Horse oocytes were recovered from abattoir ovaries by aspiration or scraping and classified as having compact (Cp), expanded (Ex), or partial (P) cumuli. In Experiment 1 (Part A in May and Part B in October), oocytes were fixed immediately after collection to assess whether the collection method influenced the initial chromatin configuration of oocytes. In Experiment 2, in vitro maturation rates of oocytes recovered by aspiration or scraping were compared. In Experiment 3, oocytes were matured in vitro and submitted to intracytoplasmic sperm injection (ICSI). Initial chromatin configuration differed according to collection method in that there was a significantly higher prevalence of diffuse chromatin within the germinal vesicle in oocytes recovered by scraping than in oocytes recovered by aspiration (29/87, 33% and 28/166, 17%, respectively; P,<,0.01). Maturation of oocytes to metaphase II did not significantly differ between scraped and aspirated oocytes (56/101, 55.4 % vs. 65/106, 61.4%, respectively). The overall pronucleus formation rate after ICSI of oocytes recovered by scraping was not significantly different than that of oocytes recovered by aspiration (50/99, 52.6% vs. 50/85, 68.5 %, respectively); however, the rate of abnormal fertilization was significantly higher for oocytes collected by aspiration (14/73, 19% vs. 6/94, 6%, respectively; P,<0.05). These results demonstrate that the collection method affects the population of recovered oocytes and may contribute to differences in results observed among laboratories working with horse oocytes. Mol. Reprod. Dev. 60: 79,88, 2001. © 2001 Wiley-Liss, Inc. [source] Infertility, infertility treatment and psychomotor development: the Danish National Birth CohortPAEDIATRIC & PERINATAL EPIDEMIOLOGY, Issue 2 2009Jin Liang Zhu Summary Babies born of infertile couples, regardless of treatment, have a higher risk of preterm birth and low birthweight, conditions associated with delayed development. We examined developmental milestones in singletons as a function of parental infertility [time to pregnancy (TTP) > 12 months] and infertility treatment. From the Danish National Birth Cohort (1997,2003), we identified 37 897 singletons born of fertile couples (TTP , 12 months), 4351 born of infertile couples conceiving naturally (TTP > 12 months), and 3309 born after infertility treatment. When the children were about 18 months old, mothers reported 12 developmental milestones by responding to structured questions. We defined a failure to achieve the assessed milestone or the minimal numbers of milestones in a summary (motor, or cognitive/language skills) as delay. Naturally conceived children born of infertile couples had a pattern of psychomotor development similar to that of children born of fertile couples, but increasing TTP correlated with a modest delay. When the analysis was restricted to infertile couples (treated and untreated), children born after treatment showed a slight delay in cognitive/language development (odds ratio 1.24, [95% confidence interval 1.01, 1.53]) for not meeting at least three out of six cognitive/language milestones); children born after intracytoplasmic sperm injection (ICSI) had the highest estimated relative risk of delay for most milestones, especially motor milestones. These results suggest that a long TTP may be associated with a modest developmental delay. Infertility treatment, especially ICSI, may be associated with a slight delay for some of these early milestones. [source] First-trimester serum marker distribution in singleton pregnancies conceived with assisted reproductionPRENATAL DIAGNOSIS, Issue 4 2010M. A. J. Engels Abstract Objective To evaluate marker distribution of free ,-human chorionic gonadotrophin (f,-hCG) and pregnancy-associated plasma protein-A (PAPP-A) in singleton pregnancies conceived by assisted reproduction techniques (ART). Methods In vitro fertilization (IVF) (n = 203) and intracytoplasmic sperm injection (ICSI) (n = 192) cases from a database of 14 645 first-trimester combined tests (overall study group) were selected and matched to 1164 controls for gestational age at sample date and maternal age. Results In the IVF group and ICSI group, lnPAPP-A was lower (IVF 6.74 vs 7.08; P = 0.0001; ICSI 6.59 vs 7.07; P = 0.0001) compared with the matched controls. Lnf,-hCG was lower in the IVF group (3.75 vs 3.90; P = 0.005) but not significantly different in the ICSI group (3.87 vs 3.93; P = 0.27). The computed correction factors for PAPP-A and f,-hCG were 1.42 and 1.17 for the IVF group and 1.56 and 1.05 for the ICSI group. The false-positive rate (FPR) in the IVF and ICSI group compared with the matched controls was higher (IVF 10.3% vs 8.6% and ICSI 10.9% vs 7.5%). In the overall age-biased [maternal age significantly lower compared with all ART and control groups] study group the FPR was 6.8%. Conclusion The increase in FPR in the ART groups can be explained by decreased PAPP-A values. Therefore, an adjustment in risk analysis for Down syndrome is suggested. Copyright © 2010 John Wiley & Sons, Ltd. [source] Full-Term Development of Rabbit Embryos Produced by ICSI with Sperm Frozen in Liquid Nitrogen without CryoprotectantsREPRODUCTION IN DOMESTIC ANIMALS, Issue 4 2010QY Li Content The aim of the present study was to establish the technology of intracytoplasmic sperm injection (ICSI) in rabbit by using the sperm frozen without cryoprotectants. Observation under an electron microscope revealed that the rabbit spermatozoa frozen without cryoprotectants had severe damage especially in the plasma membrane and junction between head and tail. However, after being injected into the oocytes, the sperm frozen without cryoprotectants retained the capability of supporting the cleavage and development of the ICSI oocytes, with no significant difference from that of fresh sperm, although the development of ICSI embryos derived from either frozen sperm or fresh sperm is much lower than that of in vivo -fertilized zygotes. When additional artificial activation was applied following ICSI, the rates of cleavage and blastocyst formation of ICSI oocytes were significantly increased when compared with the oocytes without additional activation. Yet, the cell numbers in blastocysts were not significantly different between the activation and non-activation group. After embryo transfer, four offspring were obtained from the oocytes microinjected with the sperm frozen without cryoprotectants. The technology established by this study may facilitate exploring the ICSI-based transgenic method in rabbit and broaden the application of ICSI technique in related field. [source] The Use of HOS Test to Evaluate Membrane Functionality of Boar Sperm Capacitated in vitroREPRODUCTION IN DOMESTIC ANIMALS, Issue 6 2002D Lechniak Contents The functional and structural integrity of sperm membrane are crucial for the viability of spermatozoa. The commonly used staining test (eosin + nigrosin) for assessing sperm membrane measures only its structural integrity. The hypoosmotic swelling test (HOS) originally developed for human sperm (Jeyendran et al. 1984) has been also applied to several species of domestic animals (bull, pig, horse, dog). The test enables to evaluate the functional status of the sperm membrane. The principle of HOS is based on water transport across the sperm tail membrane under hypoosmotic conditions. It has previously been used to assess the semen quality (Revell and Mrode 1994), to analyse fertilizing capacity (Rota et al. 2000; Perez-Llano et al. 2001) and also to detect viable, immotile cells for ICSI (Intra-cytoplasmic sperm injection) in human (Zeyneloglu et al. 2000). There are two procedures commonly used for sperm capacitation in the pig-sperm washing and incubation before insemination (Nagai 1994). Capacitation involves several changes like removing molecules coating the sperm head membrane, changes in membrane fluidity and intracellular ion concentration (Green and Watson 2001). Thus the membrane integrity as well as functionality may be affected as shown by Harrison (1996). The aim of the present study was to analyse changes in sperm membrane integrity after in vitro capacitation by use of the HOS test. [source] Robust computer-controlled system for intracytoplasmic sperm injection and subsequent cell electro-activationTHE INTERNATIONAL JOURNAL OF MEDICAL ROBOTICS AND COMPUTER ASSISTED SURGERY, Issue 1 2009K. K. Tan Abstract Introduction Intracytoplasmic sperm injection (ICSI) and the subsequent cell electro-activation process is a relatively new enhanced procedure to address male factor infertility. The current method involves the engagement of experienced embryologists for such a purpose. More advanced methodologies, which use high precision instrumentation tools, will speed up the whole procedure. Methods In this paper, the development of a computer-controlled system for ICSI and the subsequent cell electro-activation process is presented. The system is integrated to a microinjection workstation and piezo-actuator to perform the ICSI procedure, with vision capability to automatically position the components precisely. A micro-pump assembly is utilized for automatic medium refreshment and a heater plate assembly provides temperature control during the cell electro-activation process. The overall system is comprehensive, comprising modular functional components integrated within a hardware architecture. Results Experimental results on mice oocytes verified the effectiveness of the developed system over the current method. Conclusions Further improvements on the instrumentation tools will improve the robustness and overall performance of the developed system. Copyright © 2009 John Wiley & Sons, Ltd. [source] Practitioner Review: Outcomes for parents and children following non-traditional conception: what do clinicians need to know?THE JOURNAL OF CHILD PSYCHOLOGY AND PSYCHIATRY AND ALLIED DISCIPLINES, Issue 3 2003Susan Golombok Background: The birth of the first ,test-tube' baby in 1978 has led to the creation of family types that would not otherwise have existed. Methods: This paper examines research on the psychological outcomes for parents and children in assisted reproduction families with particular attention to the issues and concerns that have been raised by creating families in this way. Results: Parents of children conceived by assisted reproduction appear to have good relationships with their children, even in families where one parent lacks a genetic link with the child. With respect to the children themselves, there is no evidence of cognitive impairment in singleton children born at full term as a result of assisted reproduction procedures, although the findings regarding the cognitive development of intracytoplasmic sperm injection (ICSI) children are currently unclear. In relation to socioemotional development, assisted reproduction children appear to be functioning well. Conclusions: Few studies have included children at adolescence or beyond, and little is known about the consequences of conception by assisted reproduction from the perspective of the individuals concerned. In addition, there are some types of assisted reproduction family, such as families created through a surrogacy arrangement or through embryo donation, about whom little is known at all. Although existing knowledge about the impact of assisted reproduction for parenting and child development does not give undue cause for concern, there remain a number of unanswered questions in relation to children born in this way. [source] Fibroblast Growth Factor (FGF), Intracellular Adhesion Molecule (sICAM-1) Level in Serum and Follicular Fluid of Infertile Women with Polycystic Ovarian Syndrome, Endometriosis and Tubal Damage, and their Effect on ICSI OutcomeAMERICAN JOURNAL OF REPRODUCTIVE IMMUNOLOGY, Issue 2 2003M. E. Hammadeh PROBLEM: The objective of this study was to determine the concentration of fibroblast growth factor (FGF) and soluble intracellular adhesions molecule (sICAM-1) in serum and follicular fluid (FF) of polycystic ovary (PCO), endometriosis and tubal factor infertility and male factor infertility patients, and to investigate the relationship between these parameters and the outcome of intracytoplasmic sperm injection (ICSI). METHOD OF STUDY: The concentration of FGF and sICAM-1 in serum and FF were determined in patients undergoing controlled ovarian hyperstimulation (COH) for ICSI therapy for various etiology of infertility and the results of cytokines concentration and ICSI outcome were compared between the groups. Twenty patients with PCO (G.I), 17 with endometriosis (G.II), 19 with tubal damage (G.III) and 19 with male factor infertility (G.IV) were enrolled in this study. Quantitative determination of levels of FGF and sICAM-1 was performed using enzyme-linked immunosorbent assays (ELISAs). RESULTS: The FGF level in serum of PCO patients (G.I) were 4.8 ± 2.3 and in FF were 104.0 ± 39.0 pg/mL. The corresponding values in the endometriosis patients group (G.II) were 5.9 ± 3.1 and 125.4 ± 74.9 pg/mL. The concentration of FGF in tubal factor infertility group (G.III) in serum was significantly higher (P = 0.009) than those observed in the PCO group (G.I) 7.4 ± 4.5 pg/mL, whereas the concentration in FF was at the same level like the other groups investigated, 128.7 ± 75.9 pg/mL. Besides, the sICAM-1 (pg/ml) concentration in FF showed a significant difference between the groups investigated (G.I, 175.3 ± 52.8; G.II 194.4 ± 32.2; G.III 233.1 ± 54.3; and G.IV 215.1 ± 54.4 ng/mL; P = 0.003). The sICAM-1 levels in serum were not significantly different between the groups (217.0 ± 42.9; 216.3 ± 73.6; 254.8 ± 79.6; 237.56 ± 78.4 ng/ml; P = 0.267). The fertilization rate was significantly higher in G.III (66.0 ± 23.89%) in comparison to G.II (38.8 ± 33.9%; P = 0.014) or G.IV (38.7 ± 22.7%; P = 0.012). The pregnancy rates were similar in all groups (30, 35.3 and 35.0, 38.6%, respectively). CONCLUSION: Both, FGF and sICAM-1 are present in serum and FF of patients undergoing controlled ovarian hyperstimulation for ICSI therapy. The FGF concentration in serum differs significantly between the groups investigated, whereas, no significant difference could be observed in the FF concentration of FGF. On the other hand, the sICAM in serum showed no significant difference between the groups, whereas, sICAM in FF demonstrated a significant difference between the patient groups investigated. On the whole, the ICSI outcome was not related to serum or FF concentrations of FGF or sICAM-1. Therefore, the mean concentration of FGF and sICAM-1 in serum and in FF could not be used to predict the fertilization rate in an ICSI program. [source] One of These Things Is Not Like the Others: The Idea of Precedence in Health Technology Assessment and Coverage DecisionsTHE MILBANK QUARTERLY, Issue 2 2005MITA GIACOMINI Health plans often deliberate covering technologies with challenging purposes, effects, or costs. They must integrate quantitative evidence (e.g., how well a technology works) with qualitative, normative assessments (e.g., whether it works well enough for a worthwhile purpose). Arguments from analogy and precedent help integrate these criteria and establish standards for their policy application. Examples of arguments are described for three technologies (ICSI, genetic tests, and Viagra). Drawing lessons from law, ethics, philosophy, and the social sciences, a framework is developed for case-based evaluation of new technologies. The decision-making cycle includes (1) taking stock of past decisions and formulating precedents, (2) deciding new cases, and (3) assimilating decisions into the case history and evaluation framework. Each stage requires distinctive decision maker roles, information, and methods. [source] Evaluation of zeta and HA-binding methods for selection of spermatozoa with normal morphology, protamine content and DNA integrityANDROLOGIA, Issue 1 2010S. H. Razavi Summary Sperm selection parameters based on morphology and motility for ICSI might not be relevant to chromatin integrity. Thus sperm selection based on sperm characteristics has been suggested. Therefore, the aim of this study was to evaluate the efficiency of the zeta and hyaluronic acid (HA) sperm selection procedures with neat semen, for recovering spermatozoa with normal morphology, protamine content and DNA integrity in infertile men. Semen samples from 77 infertile couples were assessed during this study. Semen analysis was carried out according to World Health Organization criteria. Protamine content, DNA integrity and sperm morphology were assessed by chromomycin A3, sperm chromatin dispersion and Papanicolaou staining respectively. The results show that both HA and zeta methods were efficient to recover spermatozoa with normal morphology and protamine content. In terms of the latter parameters, there was no superiority between the two procedures. However, in terms of DNA integrity, the zeta method was more efficient compared with the control and HA procedure and no significant difference was observed between HA and the controls. Therefore, the zeta method appears to be a suitable procedure to recover spermatozoa with normal DNA integrity. [source] Fertilisation and pregnancy outcome after ICSI in globozoospermic patients without assisted oocyte activationANDROLOGIA, Issue 1 2009S. Bechoua Summary The successful outcome of intracytoplasmic sperm injection (ICSI) with globozoospermic sperm and non-activated oocytes is reported. Three couples underwent ICSI treatment and two of the patients were siblings. Forty-four non-activated oocytes were injected, 26 oocytes fertilised normally and 17 good quality embryos were obtained. Six embryo transfers were carried out, three with fresh embryos and three with frozen-thawed embryos. Three pregnancies resulted from the fresh embryo transfers and additionally two pregnancies were obtained after the transfer of frozen-thawed embryos. Two healthy babies were born. One twin pregnancy is ongoing. Our case reports demonstrate that in some ICSI attempts undertaken with globozoospermic sperm cells from two of our patients, high fertilisation rates, pregnancies and live births can be achieved, without artificially activated oocytes. Our data also suggest that in some cases, round-headed spermatozoa lack the capacity to activate the oocyte. Therefore, it cannot be excluded that artificial oocyte activation could be of help in globozoospermic patients with complete fertilisation failure. [source] Microdissection testicular sperm extraction and IVF-ICSI outcome in nonobstructive azoospermiaANDROLOGIA, Issue 4 2008P. Ravizzini Summary We evaluated the efficiency of microdissection testicular sperm extraction (MicroTESE) in patients with nonobstructive azoospermia (NOA) and their pregnancy outcomes in a programme based on in vitro fertilisation (IVF)/intracytoplasmic sperm injection (ICSI). Fifty-six MicroTESE procedures were performed in 53 patients with NOA. Pre-operative levels of luteinising hormone, follicle-stimulating hormone (FSH), testosterone and prolactin were obtained and a Doppler sonography examination was conducted. Sperm retrieval rate, mean age of female partner, mean ICSI and fertilisation rate, number and quality of embryos transferred, implantation, pregnancy and miscarriage rates were calculated. Samples for testicular histological analysis were taken trans-operatively in every case. Sperm retrieval rate, mean ICSI per case and fertilisation rate were 57.1%, 7.4% and 58.4% respectively. A significant difference in pre-operative testicular volume (P = 0.001), serum FSH (P = 0.008) and total testosterone levels (P = 0.021) was found in patients from whom sperm could be retrieved. Mean 1.9 type A embryos were transferred per cycle. Implantation, clinical pregnancy and miscarriage rates were 20%, 40% and 18.7% respectively. It is concluded that MicroTESE is a viable option for men with NOA, offering excellent results in couples undergoing IVF-ICSI. Pre-operative serum FSH, testicular volume and total testosterone levels may have a prognostic value, although more data are needed to determine their significance and whether or not patients should be excluded from an initial sperm retrieval attempt. [source] Primary male infertility in Kuwait: a cytogenetic and molecular study of 289 infertile Kuwaiti patientsANDROLOGIA, Issue 3 2007F. Mohammed Summary Infertility is one of the major public health problems, affecting 15% of couples who attempt pregnancy; in 50% of these, the male partner is responsible. Chromosomal abnormalities and Y microdeletions in the azoospermia factor (AZF) region are known to be associated with spermatogenetic failure. In the present study, 289 patients with primary male infertility because of spermatogenetic failure were studied in order to highlight the molecular background of male infertility in Kuwait, and to avoid the possibility of transmission of any microdeletions/chromosomal aberrations to offspring via intracytoplasmic sperm injection (ICSI). Of the 289 infertile men, 23 patients (8%) had chromosomal aberration in the form of Klinefelter syndrome/variant (16/23; 69.6%), XYY syndrome (3/23; 13%), XX male syndrome (2/23; 8.7%), 45,X/46X, i(Yp)(1/23; 4.4%) and 45,XY, t(9;22) (1/23;4.4%). Y-chromosome microdeletion in the AZFb and AZFc regions were detected in 7/266 cases (2.6%). Testicular biopsy was carried out in 31 azoospermic patients, of whom five men had Sertoli-cell only syndrome, while 26 patients had spermatogenic arrest. In conclusion, this study showed that the frequency of both chromosomal anomalies and Y microdeletions were found in 10.4% of the infertile men. The potential risk of transmitting these genetic disorders to offspring provides a rationale for screening infertile men prior to ICSI. [source] Influence of motility and vitality in intracytoplasmic sperm injection with ejaculated and testicular spermANDROLOGIA, Issue 4 2005T. Stalf Summary The vitality of spermatozoa used for intracytoplasmic sperm injection (ICSI) is a crucial factor for fertilization, establishment and outcome of a pregnancy in assisted reproductive technique cycles. The sperm origin may also be a limiting factor, although little is known about this issue. It is known that the motility of injected spermatozoa and their origin from ejaculate or testicular biopsies are important predictors in terms of fertilization, pregnancy and birth rates. Oocytes of patients in 2593 cycles were retrieved in our in vitro fertilization programme and inseminated via ICSI. We used motile (group 1, n = 2317) or immotile ejaculated spermatozoa (group 2, n = 79), motile sperm retrieved from testicular biopsies (group 3, n = 62) and immotile spermatozoa from testicular biopsies (group 4, n = 135). Female age and number of oocytes retrieved did not differ significantly among the groups. The fertilization rates were as follows: 67.1% in group 1, 49.8% in group 2, 68.3% in group 3 and 47.8% in group 4. The pregnancy rates in cases where three embryos had been transferred amounted to 35.7% in group 1, 17.3% in group 2, 38.3% in group 3 and 20.5% in group 4. The embryo quality showed no differences between groups 1 and 3 (14.5), and between groups 2 (11.8) and 4 (10.8). The abortion rate was similar in groups 1,3, but increased in group 4 (26.6%, 27.3%, 31.6% and 55.5%). Irrespective of their origin, the fertilization potential of injected spermatozoa was found to be influenced by motility. The resulting pregnancy and birth rates, i.e. the potential of the resulting embryos to implant and to achieve viable pregnancies, seem to be additionally dependent on the sperm origin. This was well shown by declining rates when spermatozoa in a relatively early stage of maturity had been used. We see increasing evidence that the degree of sperm maturity has an important impact on the outcome of ICSI. In obstructive azoospermia, spermatozoa retrieved from the epididymis should be used rather than testicular biopsy spermatozoa, or testicular sperm should be preincubated in culture medium before ICSI. [source] Effects of paternal cigarette smoking on testicular function, sperm fertilizing capacity, embryonic development, and blastocyst capacity for implantation in ratsANDROLOGIA, Issue 2 2004A. Kapawa Summary. We evaluated the effects of paternal smoking on testicular function, sperm fertilizing capacity, embryonic development, and blastocyst capacity for implantation. Rats of group A were exposed to cigarette smoke for 10 weeks. Rats of group B were exposed to the smoke of incense sticks for 10 weeks. Rats of group C served as a control group. Rats of group D were exposed to cigarette smoke for 7 weeks only. Experimental period was 10 weeks in all groups. At the end of the experimental period serum testosterone responses to human chorionic gonadotropin stimulation, andro-gen-binding protein activity in testicular cytosols, epididymal sperm motility, and oocyte fertilization rate, oocyte cleavage rate, and blastocyst development rate after in vitro fertilization (IVF) trials were significantly smaller in group A compared with groups B and C. In contrast, fertilization rate, cleavage rate, and blastocyst development rate after intracytoplasmic sperm injection (ICSI) procedures were not significantly different among groups A, B, C, and D. Both after IVF trials and ICSI techniques, the proportion of the alive offspring to the number of transferred oocytes was significantly smaller in group A than in groups B and C. Cigarette smoke-exposure results in a secretory deficiency of Leydig and Sertoli cells leading to an impaired epididymal sperm maturation process and diminished capacity of spermatozoa to penetrate oocytes. In addition paternal cigarette smoke exposure affects the embryonic ability for implantation. [source] Morphological and morphometric attributes of epididymal and testicular spermatozoa following surgical sperm retrieval for obstructive and nonobstructive azoospermiaANDROLOGIA, Issue 6 2003Dr. S. Wood Summary. Whilst the morphological (shape) and morphometric (sperm head size) attributes of ejaculated spermatozoa have been well studied, the morphological and morphometric qualities of testicular and epididymal spermatozoa retrieved from males with obstructive and nonobstructive azoospermia is much less documented. We wished to examine the effect of aetiology of azoospermia and site of retrieval on the attributes of retrieved spermatozoa. This was a prospective observational study of 30 consecutive successful sperm retrievals, six for nonobstructive azoospermia and 24 for obstructive, of which five were retrieved from the epididymis and the remainder from the testis. The proportion of morphologically normal testicular spermatozoa in patients with obstructive and nonobstructive azoospermia was not significantly different (7% versus 7.6%, P = 0.97). Testicular spermatozoa from males with obstructive azoospermia showed an increase in frequency of sperm with small heads [47/180 (26%) versus 97/909 (11%), P = 0.036] as well as small acrosome and increasing vacuole formation over nonobstructive spermatozoa. Similarly, there was a significant increase in tail deformities and decreases in tail lengths in sperm from males with nonobstructive azoospermia. Epididymal spermatozoa showed significantly greater proportion of morphologically normal spermatozoa than testicular (20% versus 13%, P = 0.001) as well as a significant increase in acrosome vacuoles. Furthermore, morphometrically epididymal spermatozoa displayed with smaller head length, width and area than testicular spermatozoa. Testicular spermatozoa from obstructive azoospermia displayed significantly less tail defects (35% versus 57%, P = 0.003) as well as significantly longer tail lengths (30.6 ,m versus 10.7 ,m). These morphological and morphometric differences between epididymal and testicular and obstructive and nonobstructive spermatozoa may represent part of the natural maturation process. There were no associations between any morphological or morphometric abnormality with any significant parameter in subsequent use in ICSI. [source] Possible predictive factors for ICSI?ANDROLOGIA, Issue 4 2003Molecular biology techniques in combination with therapeutic testicular biopsies Summary. Applying intracytoplasmic sperm injection (ICSI), the selection of an unsuccessful spermatozoon results in great emotional consequences for the couple. Therefore, there is a need for a prognostic parameter to estimate their chances for successful fertility treatment. This review summarizes both the main reasons for spermatogenic impairment, and possible predictive factors for successful sperm retrieval applying testicular sperm extraction and outcome of ICSI. While basic sperm parameters, aetiology and type of spermatozoa, and serum follicle-stimulating hormone and inhibin levels have been shown to be unrelated to the outcome of ICSI, Y-chromosome microdeletions are known to have a negative influence on the fertilizing capacity of spermatozoa. Recently, a significant correlation has been reported between the protamine-1 to protamine-2 mRNA ratio in haploid spermatids of testicular biopsies and the ability of spermatozoa for successful fertilization of an oocyte. In future, both the outstanding role of the haploid spermatids and the involvement of molecular biological techniques will improve the role of therapeutic testicular biopsies. [source] Better sperm selection for intracytoplasmic sperm injection with the side migration techniqueANDROLOGIA, Issue 6 2001A. Hinting Summary. The side migration technique (SMT) is a recent method for preparing very poor-quality semen samples to be used in intracytoplasmic sperm injection (ICSI). In most centres, the washing swim-up and Percoll gradient columns techniques have been routinely used. The present study is aimed at comparing the quality of oligozoospermic semen samples selected after these three methods. All three methods were found to select better percentage motility, normal morphology, viability, functional integrity of plasma membrane and nuclear chromatin integrity compared with the original semen samples. Among the three methods, however, SMT yielded better sperm quality, including morphology, viability, membrane integrity and nuclear chromatin integrity. The results of this study and our experience have confirmed that SMT is an effective and physiological method to prepare sperm for ICSI. [source] Embryo development of porcine oocytes after injection with miniature pig sperm and their extractsANIMAL SCIENCE JOURNAL, Issue 6 2009Daizou MATSUURA ABSTRACT This study examined embryo development of porcine oocytes after microinjection of sperm extracts (SE) in porcine intracytoplasmic sperm injection (ICSI). SE was prepared from miniature pig sperm by a nonionic surfactant, and various concentrations (0.02, 0.04 and 0.08 mg/mL) of SE were injected into the matured oocytes with a first polar body. In the pronuclear stage, the rate of oocytes with two pronuclei and a second polar body (21.4%) in the sperm and SE (0.04 mg/mL) injection group was significantly higher (P < 0.05) compared to other groups. The rate of 2,4-cell stage in sperm and SE (0.04 mg/mL) injection group was 38.1%, and it was significantly higher than that in the sperm injection group (22.9%). The rate of blastocyst stage in sperm and SE (0.04 mg/mL) injection group was 21.4%, the value was significantly higher than those in SE (0.08 mg/mL) injection group (0%), sperm injection group (5.7%), and sperm and SE (0.08 mg/mL) injection group (2.6%). These results suggest that SE induces activation of porcine oocytes and their further embryonic development, and that SE is effective for porcine ICSI. [source] | ||||||||||||||||||||||||||||||||||