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[Assisted fertilization--where are we now and where are we going?].

The authors review the development of assisted reproduction, and describe the present status. The efficacy of in vitro fertilization is compared with the results of tubal surgery. A comparison is also made of the efficacy of different assisted reproductive techniques. Future aspects of assisted reproduction are discussed, with particular emphasis on the need for better understanding of the implantation process. This may reduce the need for transfer of multiple embryos, thereby decreasing risk of multiple pregnancies.

Female↗

Negative effects of increased sperm DNA damage in relation to seminal oxidative stress in men with idiopathic and male factor infertility.

OBJECTIVE: To examine the effects of increased sperm DNA damage in relation to seminal oxidative stress in men with idiopathic and male factor infertility. Prospective study. SETTINGS: Infertility clinic at a tertiary care academic institution. PATIENT(S): Ninety-two infertile men with normal female partners. Sixteen fertile donors served as the control group. INTERVENTION(S): Standard semen analysis and assessment of levels of seminal oxidative stress. Assisted reproductive techniques in 33 of the 92 patients (IUI [n = 19], IVF [n = 10], and intracytoplasmic sperm injection [n = 4]). MAIN OUTCOME MEASURE(S): Sperm DNA damage by sperm chromatin structure assay. Results were expressed as DNA fragmentation index. RESULT(S): Patients were classified as having either idiopathic (n = 23) or male factor infertility (n = 69). Patients with idiopathic and male factor infertility had significantly higher DNA fragmentation index and oxidative stress compared with the case of fertile donors. A clinical pregnancy was achieved in 9 (27%) of 33 patients with assisted reproductive techniques. Significantly higher DNA fragmentation index and oxidative stress were found in men who failed to initiate a pregnancy after assisted reproductive techniques (n = 24), compared with the cases of those who succeeded and of the fertile donors. DNA fragmentation index was correlated positively with oxidative stress (r = 0.27), and negatively with fertilization (r = -0.70) and embryo quality (r = -0.70). CONCLUSION(S): Sperm DNA damage is significantly increased in men with idiopathic and male factor infertility and in men who failed to initiate a pregnancy after assisted reproductive techniques. Such an increase may be related to high levels of seminal oxidative stress.

Adult↗

Prospects for human reproductive medicine--applications of experimental techniques used in farm animal breeding.

Can assisted reproduction techniques performed in farm animal breeding be considered as future methods in human sterility treatment? The answer is no because, at the moment, the goals in human and animal reproduction are totally different. The methods that scientists are using or developing for farm animals are either without any therapeutic value for humans or they are forbidden by the German Embryo Protection Law. However, there are a few aspects which might be of value in the future of human assisted reproduction. The technique of in vitro maturation of immature follicles and oocytes, which is prerequisite in animal in vitro fertilization the female gamete pool after slaughter, might be used to obtain mature human oocytes from unstimulated cycles in women who do not respond to hormonal stimulation. Further more, it can be used in cancer patients to save their oocytes shortly before cancer treatment. Although still forbidden at the moment by the Embryo Protection Law, it is possible that the pre-implantation diagnosis of early embryos by the polymerase chain reaction technique after blastomere biopsy will become a valuable tool in preventing genome aberration in future.

Animals↗

Is there a possible correlation between chromosomal variants and spermatogenesis?

AIM: Heterochromatin polymorphism is considered a variant of a normal karyotype but is more frequent in infertile men. The aim of this study was to evaluate the correlation between heterochromatic variants and male infertility and to discuss the possible mechanisms of how heterochromatic polymorphism might affect spermatogenesis. METHODS: Cytogenetic analysis was undertaken in 210 infertile males who had been taken into assisted reproductive techniques and in 183 men with proven fertility. Additionally, C-banding was performed in men with heterochromatin polymorphism. Sperm fluorescence in situ hybridization (FISH) was applied in 54 men with normal karyotype presenting either normal or abnormal sperm parameters as well as in 8 men with heterochromatin polymorphism. The outcomes of assisted reproductive techniques were compared between infertile men with normal karyotype and men with heterochromatin polymorphism. RESULTS: The incidence of heterochromatin polymorphism was higher in infertile men. The most frequent chromosome involved in heterochromatin polymorphism was chromosome 9. Sperm FISH analysis revealed an increased rate of aneuploidy in men with heterochromatin polymorphism. Laboratory and clinical outcomes in assisted reproductive techniques were compromised in men with heterochromatin polymorphism. CONCLUSIONS: An increased rate of heterochromatin polymorphism in infertile males seems to be more than an incidental finding, and must not be considered as a normal variant. Polymorphic heterochromatin may have deleterious effects on the genetic constitution of spermatozoa. More attention must be directed to infertile men with heterochromatin polymorphism.

Adult↗

Ethical issues in modern biological technologies.

Today's biology-based technologies have emerged from a historical imperative and as an inevitable consequence of developments in modern biology beginning in the last half-century. They can be classified into almost 30 different areas, ranging from the use of gene therapy for human beings, enzyme engineering, stem cells and cloning, to marine biotechnology, bioinformatics, nanotechnology and biological warfare among many others. Many of them have major sociopolitico-economic, moral, ethical and legal implications. They include genetic engineering, gene therapy, tissue culture, stem cell work, the new DNA technologies, commercialization of traditional plant-based drug formulations, assisted reproduction techniques, cloning technologies, organ transplantation, bioinformatics, and biological weapons. Examples of the ethical implications of several of these items will be considered. They will be assessed with special reference to ethical implications in respect of assisted reproduction techniques, of worldwide importance today, particularly for a country such as India.

Biological Warfare↗

Investigation of fertilizing capacity of cryopreserved spermatozoa from patients with cancer.

PURPOSE: There are few published reports concerning fertilization and pregnancy outcomes achieved with cryopreserved spermatozoa from cancer patients. Controversy exists regarding the value of sperm banking for these patients before therapy, whether the spermatozoa are viable after long-term storage and whether they can fertilize the ovum. We assess fertilization and pregnancy outcomes achieved with cryopreserved spermatozoa from cancer patients using assisted reproductive techniques. MATERIALS AND METHODS: We studied 10 cancer patients who transferred cryopreserved semen specimens from our sperm bank to outside in vitro fertilization programs for assisted reproductive technique. Of these patients 5 had Hodgkin's disease, 2 testicular cancer, 1 leukemia and 2 prostate cancer. The length of specimen storage ranged from 14 to 135 months (median 49, interquartile range 24 and 82). RESULTS: The median pre-freeze motility was 44% (interquartile range 36 and 55%) and the median total sperm count was 31.1 x 10(6) (interquartile range 6.3 and 53.9 x 10(6)). At 24 hours after banking the median post-thaw motility was 11% (interquartile range 6 and 35%) and the median total sperm count was 6.6 x 10(6) (1.2 and 17.1 x 10(6)). A total of 18 cycles of assisted reproductive technique were performed among 10 couples with an overall pregnancy rate of 50% per couple, with 2 deliveries, 1 ongoing pregnancy and 2 miscarriages. The pregnancy rate per cycle of in vitro fertilization and intracytoplasmic sperm injection was 36.4% with an implantation rate of 13%. CONCLUSIONS: These results indicate that poor quality cryopreserved spermatozoa from cancer patients, irrespective of the length of storage, may provide successful results with the latest micromanipulative techniques such as intracytoplasmic sperm injection.

Adolescent↗

Experience with vasal sperm aspiration.

PURPOSE: We describe our experience with vasal aspiration of sperm in patients with azoospermia secondary to neurogenic anejaculation or obstruction of the vas deferens. MATERIALS AND METHODS: We performed 15 vasal sperm aspirations on 11 patients with neurogenic anejaculation or vasal obstruction. RESULTS: Adequate sperm was obtained from all 15 aspirations with an average sperm count of 42 x 10[6] (range 0.5 to 252 x 10[6]) and an average motility of 73.4% (range 16 to 100%). Aspirations were performed for use with assisted reproductive techniques in 12 cases and for cryopreservation alone in 3. Following assisted reproductive techniques 6 of the 12 aspirations (50%) resulted in pregnancy. CONCLUSIONS: Vasal aspiration of sperm should be considered when electroejaculation fails, is not available or is contraindicated. Although the overall sperm quality and pregnancy rate are higher with vasal aspiration and in vitro fertilization than electroejaculation and intrauterine insemination, vasal aspiration is a more technically demanding and costly approach to conception.

Adult↗

Filiation is assisted reproduction: potential conflicts and legal implications.

Filiation, or a person's parentage, is one of the major components of someone's identity, often with important implications for that person's welfare in social and personal terms. Fertility specialists are enjoined to consider the welfare of the child, and this is a legal requirement in assisted reproduction techniques in the UK. This paper is in three parts, discussing some of the potential problems regarding filiation of the prospective child born of assisted reproduction techniques, in relation to the use of gametes and embryos. The first two parts concentrate on the status of children born as a result of treatment given posthumously, and on filiation problems associated with donation of gametes or embryos, and surrogacy. The third part compares British and French legislation with special regard to the welfare of the child, as well as underlining the common trends and principles enunciated in both legislations. This is particularly appropriate at a time when the European Society for Human Reproduction and Embryology is about to issue ethical guidelines in assisted reproduction, and when both the Council of Europe and the European Commission have commissioned studies on the implications of the assisted techniques, including those concerning status of the potential children.

Child Welfare↗

Assisted reproductive research: laser assisted hatching and spindle detection (spindle view technique).

Animal experiments are very important for the development of new assisted reproductive techniques (ART) for use in human and animal reproductive medicine. Most technical aspects of reproductive manipulation of humans and animals are very similar, and many components of successful human ART used nowadays have been derived from animal studies. In this study we examined (1) the use of 'non-contact' laser for assisted hatching, (2) whether spindles in living mouse oocytes could safely be imaged/examined by polarisation microscope (polscope) and (3) the influence of environment (e.g. temperature, in vitro culture, etc.) on spindle detection/visualisation. The data of the study presented here show that (1) laser assisted hatching (AH) is a fast, very accurate and safe procedure without any harmful effect on embryo development and it can support very effectively the implantation of embryos, (2) the use of polscope facilitates the evaluation of oocyte quality and the selection of oocytes with spindle, (3) by monitoring the spindle position during intracytoplasmic sperm injection (ICSI), we can reduce spindle damage and increase the chance of fertilisation. Further studies are underway to test the hypothesised connection between spindle birefringence and developmental capacity of oocytes/embryos.

Animals↗

The European study of assisted reproduction families: a comparison of family functioning and child development between Eastern and Western Europe.

Developments in reproductive technology over the past 20 years have given infertile couples new opportunities to have children. Although assisted reproduction does not appear to lead to negative consequences with respect to parent-child relationships or the socioemotional development of the child, the outcome for children and their parents may well differ according to the social and cultural environment. The aim of the present study was to compare family functioning and the social and emotional development of children in families created as a result of assisted reproductive techniques in Western Europe with assisted reproduction families in an Eastern European country (Bulgaria) where there is a history of specific pronatalist interventions. The findings indicate greater difficulties in parental adjustment and child behavior in assisted conception families in Eastern Europe. In addition, Eastern European parents were more uncertain than Western European parents about whether they would tell the child, and Eastern European parents were much more secretive with respect to telling others. Thus the outcomes of assisted reproduction for family functioning and child development appear to be independent, to some extent at least, on the social context in which these techniques are carried out.

Adaptation, Psychological↗

Collection, evaluation, and use of oocytes in equine assisted reproduction.

Assisted reproductive techniques have been developed to obtain pregnancies from subfertile mares and stallions and to salvage gametes after death. In recent years, these procedures have been used for clinical cases with repeated success. Although new developments occur, the basis for the success and future development of assisted reproductive techniques is our ability to collect and handle the equine oocyte successfully. This article focuses on important clinical aspects of oocyte collection and evaluation and briefly discusses the clinical use of assisted reproductive procedures in the horse.

Animals↗

Possibilities and limitations of techniques of assisted reproduction for the treatment of male infertility.

Since relatively few spermatozoa are needed for oocyte fertilization during gamete intra-Fallopian transfer (GIFT) or in-vitro fertilization (IVF), these methods have been applied in couples with infertility due to male causes. Forty-six couples with male factor infertility were enrolled in this study and results were compared with those attained in 48 couples treated with the same techniques for other than male causes. Overall, GIFT resulted in 26% ongoing pregnancies. GIFT seems to be particularly successful when the sperm concentration is 20 x 10(6)/ml or more, but sperm motility and/or morphology are poor. Nine pregnancies occurred out of 26 GIFT cycles in 18 cases selected on this basis. The ongoing pregnancy rate after IVF was 16% per patient. The latter treatment should be attempted in male immune infertility and in cases with a low sperm concentration, with or without abnormal sperm motility and/or morphology. In these circumstances, five pregnancies were attained out of 28 cycles in 14 cases. For similar sperm concentrations, the conception rate per cycle attained with techniques of assisted reproduction was more than twice that attained with conventional treatment of male infertility.

Female↗

Serum hormones for predicting pregnancy outcome after assisted reproductive technology.

Serum human chorionic gonadotrophin (HCG) in the second and third week after embryo transfer has been used for prediction of pregnancy outcome after assisted reproduction. There are few data on the clinical utility of HCG, progesterone and oestradiol, measured by contemporary immunoassay, in the fourth week after embryo transfer and later. Moreover, large inter-method differences have been described between automated immunoassays, making method-specific cut-off values mandatory. The main aim of this study was to determine assay-specific optimal cut-off values for serum HCG, progesterone and oestradiol for prediction of clinical pregnancy outcome in singleton pregnancies after assisted reproductive techniques, at days 11, 18 and 25 and at week 6 after embryo transfer. A retrospective study was performed on frozen serum samples of 67 singleton pregnancies after assisted reproduction techniques. HCG, oestradiol and progesterone were determined with the automated (random access) VIDAS immunoanalyser. Receiver operating characteristic curve analysis was performed to determine optimal cut-off values. Predictive values were calculated based on the prevalence of non-viable pregnancy after assisted reproduction. It was concluded that measurement of HCG by VIDAS at days 18 and 25, and at week 6 after embryo transfer yields high positive (70.5-100%) and negative (87.2-94.4%) predictive values for clinical pregnancy outcome.

Chorionic Gonadotropin↗

[Management of twin births].

INTRODUCTION: The rate of multiple births has increased all over the world in the past decade. This is mainly due to the widespread use of infertility treatments and assisted reproductive techniques. Despite of improvements in obstetric and neonatal care, twin pregnancy still mean high risk both for the mothers and their babies. The management of twin delivery remains controversial. PATIENTS AND METHOD: Authors analyzed the data of all twin pregnancies at the Department of Obstetrics and Gynecology, University of Debrecen between January 2002 and December 2003. The data of 113 twin pregnancies were analyzed. Details of gestational age, presentation, birth weight discordance, mode of delivery were reviewed. RESULTS: From the 113 analyzed twin births, 27 pregnancies have been conceived after successful assisted reproductive technique. In 18 cases more than 500 grams of body weight difference was detected between the two babies at the time of delivery. The rate of preterm birth was 69%, and cesarean section was performed in 64% of preterm births compared to 68.6% in the matured delivery group. CONCLUSIONS: On the basis of recent data a large prospective, randomized trial is mandatory to determine the optimal management of twin deliveries.

Cesarean Section↗

[Assisted reproduction technologies and the risk of fetal, chromosomal and genetic malformations].

BACKGROUND: Assisted reproduction techniques allowed thousands of otherwise infertile couples to attain pregnancy. As this technology moves into the mainstream of infertility treatment, it has become more critical to reassess its safety. OBJECTIVE: To review the birth outcome of patients undergoing conventional in-vitro fertilization and intracyto- plasmic sperm injection regarding fetal malformations, chromosomal and genetic abnormalities. METHODS: Selective review of the literature. RESULTS: Most of the published data is from observational studies and is not randomized or blinded. Unfortunately, most articles are inherently biased. Chromosomal and genetic abnormalities are increased probably only as a direct corollary to the underlying parental risk and not due to the technology itself. There is a slight increase in the congenital malformations rate, but inspection of these malformations reveal no clustering of any specific abnormality. CONCLUSIONS: Children born after assisted reproduction technologies have an increased risk of a major congenital malformation and chromosomal abnormalities compared with those born after natural conception. The risk is mainly due to paternal and maternal risk factors, which are more prevalent in couples who use assisted reproduction techniques for reproduction. Infertility-linked risk is highly probable for the observed findings. A technique-related risk, however, cannot be ruled out. Intracytoplasmic sperm injection appears to be a safe alternative for couples who otherwise would be unable to achieve pregnancy. The inherent risks associated with these genetically "at risk" couples mandate thorough evaluation and counseling before undertaking ICSI.

Congenital Abnormalities↗

Serum insulin-like growth factor-I (IGF-I) and growth in children born after assisted reproduction.

CONTEXT: Concern has been raised about the safety of assisted reproduction techniques for the offspring. OBJECTIVES: The objective of the study was to investigate postnatal growth and growth factors in children born after intra-cytoplasmatic sperm injection (ICSI) and in vitro fertilization (IVF). DESIGN: The study had two cohorts: a population-based longitudinal infant cohort 0-36 months [236 ICSI, 173 IVF, 1530 naturally conceived (NC)], and a cross-sectional child cohort at 5 yr (68 ICSI, 67 IVF, 70 NC). INTERVENTION: Anthropometrical measurements were made at birth, 3, 18, 36 (infant cohort), and 60 months (child cohort), and blood samples were collected at 3 or 60 months. MAIN OUTCOME MEASURES: Serum IGF-I, IGFBP-3, height, weight, head and abdominal circumference, body mass index, and fat folds were the main outcome measures. RESULTS: Anthropometrical measurements showed no significant differences between ICSI and IVF children and controls in either cohort. However, singleton ICSI girls [3.4 (0.6) kg, P = 0.008] had a slightly lower birth weight than IVF [3.5 (0.5) kg] and NC girls [3.5 (0.5) kg]. Birth weights of singleton boys [3.6 (0.5) kg], twin boys [2.6 (0.6) kg], and twin girls [2.4 (0.5) kg] did not differ between types of conception. In the infant cohort in 3-month-old singletons, serum IGF-I was lower in ICSI [78 (26) ng/ml] than NC boys [94 (27) ng/ml, P < 0.001] and IVF [74 (34) ng/ml], compared with NC girls [93 (43) ng/ml, P = 0.011]. ICSI children were also smaller than their target height (sd score) at 3 yr of age [mean -0.91 (1.2)], compared with NC children [-0.61 (0.9), P = 0.033]. In the child cohort, target height attainment (sd score) and growth factors did not differ among the three groups. CONCLUSIONS: The overall growth pattern of ICSI and IVF children in both cohorts was normal. Our findings of subtle differences in target height attainment and serum IGF-I levels between infants born after assisted reproduction techniques and controls may not be clinically significant. However, these observations indicate that further systematic follow-up of growth and puberty in these children is needed.

Body Height↗

Successful pregnancy following zygote intrafallopian transfer for congenital cervical hypoplasia.

BACKGROUND: Congenital cervical atresia and hypoplasia are rare abnormalities that generally require reconstructive or extirpative procedures to relieve outflow tract obstruction. Infertility is a common sequel, and only four previous pregnancies have been reported. In selected cases, zygote intrafallopian transfer (ZIFT) or other assisted reproductive techniques may offer alternatives for conception. CASE: A 21-year-old amenorrheic woman experienced a spontaneous gush of vaginal bleeding following an 11-year history of cyclic lower abdominal pain. Regular but prolonged and painful menses ensued. After another 8 years of primary infertility, transcervical and transfundal hysteroscopy demonstrated congenital cervical hypoplasia and a normal endometrial cavity. Conception was achieved during her third cycle of ZIFT. Delivery occurred by elective cesarean at 39 weeks for a persistent oblique fetal lie. CONCLUSION: A successful pregnancy was established following ZIFT in a woman with congenital cervical hypoplasia. The endometrial cavity was evaluated by a previously unreported technique, transfundal hysteroscopy. The use of appropriate surgical or assisted reproductive techniques in conjunction with individualized post-conception management may permit successful pregnancy and delivery in selected women with congenital cervical hypoplasia and atresia.

Adult↗

Luteal phase support.

Assisted reproductive techniques have become a routine treatment for infertility. The extended use of gonadotrophin-releasing hormone analogues in assisted reproductive techniques has made luteal phase support mandatory, as it has been clearly demonstrated that they alter luteal LH pulsatility. For luteal support, HCG administration, though effective, has a high risk of ovarian hyperstimulation syndrome. Progesterone continues to be the gold standard for supplementation. Vaginal progesterone represents a highly effective alternative to painful intramuscular injections. The vaginal route is mainly characterized by direct delivery of the progesterone to the endometrium, thus producing high levels at the target tissue and a very low incidence of side effects.

Journal Article↗