ICSI diagnostic: a way to prevent total fertilization failure after 4 unsuccessful IUI

Background

The aim of this retrospective study is to investigate the relevance of dividing oocytes and using some for traditional in vitro fertilization (IVF) and others for intracytoplasmic sperm injection (ICSI) as of the first IVF cycle in patients with unexplained infertility who have undergone 4 intrauterine insemination (IUI) cycles which produced no pregnancies.

Methods

This retrospective study includes patients with unexplained infertility who have failed to become pregnant, after 4 IUI, despite normal semen parameters after sperm capacitation. These women were treated in our assisted fertilization program from 2008 until 2015. We analysed the first cycles of women in whom more than 4 oocyte cumulus complexes (OCC) were retrieved and single embryo transfer was performed.

Results

Dividing oocytes between two fertilization techniques reduce the rate of total fertilization failure during the first IVF cycle. No statistical difference were observed for 2 pronuclei (PN) rate between the two techniques. On the other hand, we observed a significantly lower rate of 3 PN, 1 PN, 0 PN with ICSI in comparison with conventional fertilization.

Conclusions

Splitting the oocytes between classical IVF and ICSI increases the chance of embryo transfer on a first IVF cycle after 4 unsuccessful IUI cycles. This half-and-half policy reduces the risk, for the infertile couple, of facing total failure of fertilization and also can provide useful information for the next attempts.

     

Total fertilization failure and idiopathic subfertility

Background  

To gain more insight in whether failure of intrauterine insemination (IUI) treatment in patients with idiopathic subfertility could be related to diminished fertilization, the aim of this study is to compare the fertilization of an initial IVF procedure after six cycles of IUI and the fertilization of an initial IVF procedure without preceding IUI cycles in couples with idiopathic subfertility.     

Genetische Risiken der assistierten Reproduktion

Infertility patients have to be counseled about the genetic risks before treatment. Chromosomal anomalies can be found in 5% of subfertile men and in cases of azoospermia the frequency even rises to 15%. Therefore a chromosome analysis should be performed in men with a sperm count <20 million/ml and also in their partners. Klinefelter’s syndrome as well as a Robertsonian translocation can cause male subfertility. Mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene and deletions in the Y chromosome have been described as being more frequent in these cases. Regarding pregnancy and neonatal outcome of children, patients have to be counseled about the increased risk of complications. The abortion rate is increased by 1.3-fold, although this is most likely caused by the subfertility of the couple and other risk factors and not caused by the therapy. Complications during pregnancy, such as preeclampsia, growth retardation, stillbirth and a low birth weight are more common after assisted reproduction. The incidence of congenital anomalies is increased by 1.3-fold after in vitro fertilization (IVF) as well as after intracytoplasmic sperm injection (ICSI) therapy.     

The Use of In Vitro Fertilization in the Management of Male Infertility: What the Urologist Needs to Know

Infertility affects approximately 10% to 20% of reproductive-age couples, many of whom may present initially to a urologist. Some couples may be treated medically to increase spontaneous conception rates; however, many will require more aggressive management with in vitro fertilization (IVF) and/or intracytoplasmic sperm injection (ICSI). IVF involves ovarian stimulation, oocyte retrieval, and fertilization outside of the body; ICSI involves injecting one sperm into the oocyte to promote fertilization. Here we provide a brief overview of IVF and ICSI along with a discussion of the risks involved to facilitate the counseling and care of the infertile couple.Key words: Intracytoplasmic sperm injection, Male infertilityInfertility, defined as the inability to conceive within 12 months of unprotected intercourse, affects approximately 10% to 20% of reproductive-age couples.¹ As couples defer childbearing until later ages and as the obesity epidemic grows, the incidence of infertility is likely to continue to rise.^2,3 Male factor infertility is estimated to contribute to two-thirds of all cases. Of men seeking care for infertility, 18.1% reported being diagnosed with male factor infertility and 13.7% with a sperm or semen problem.⁴The evaluation for male infertility includes a thorough history and physical examination, and the mainstay of diagnostic testing continues to be the semen analysis. If abnormalities are noted on semen analysis, further testing is warranted to evaluate for possible etiologies. Where applicable, treatment is initiated with the goal of improving semen quality and male fertility. Previously, in cases in which semen quality remained profoundly impaired, the successful treatment for male factor infertility was once limited to donor insemination.The development of in vitro fertilization (IVF) revolutionized the management of female infertility. As powerful a tool as this proved to be, however, IVF fertilization rates remained poor in the presence of compromised semen parameters. A significant breakthrough in the treatment of severe male infertility was the development of intracytoplasmic sperm injection (ICSI) in 1992.⁵ By allowing the injection of a single sperm into each oocyte, ICSI provides the possibility of genetic offspring to men who have very scant numbers of motile sperm on semen analysis or who require surgical harvesting.From its inception, assisted reproduction has involved a gynecologist and an embryologist. The urologist is a critical collaborator for the treatment of couples with male factor infertility. Sperm harvested by microsurgical epididymal sperm aspiration, testicular sperm aspiration, or biopsy can be used to fertilize harvested oocytes by ICSI. The urologist may be the first to evaluate a couple for infertility, and will certainly be involved if sperm harvesting is indicated. Therefore, this article reviews the process of assisted reproduction by IVF/ICSI for urologists who may be seeing patients with infertility issues.     

Tracking microdeletions of the AZF region in a patrilineal line of infertile men

Male infertility is considered to be a difficult-to-treat condition because it is not a single entity, but rather reflects a variety of different pathologic conditions, thus making it difficult to use a single treatment strategy. Structural alterations in the Y chromosome have been the principal factor responsible for male infertility. We examined 26 family members of 13 patients with male infertility who showed deletions in the AZF region. In family 1, the father and a brother did not show microdeletions. However, a son showed a microdeletion in AZFa (sY84) and an azoospermic sperm analysis, but another son had a microdeletion in AZFa (sY84) and AZFb (sY127) and a normal sperm analysis. The father of family 2, with severe oligozoospermia, had a microdeletion in the AZFa region (sY84) and his son, conceived by intracytoplasmic sperm injection, also showed the same microdeletion. In the other families, only the men with an altered sperm analysis had a microdeletion. It is possible that in family 1, the father and brother who did not show microdeletions in this study, could have microdeletions in regions upstream or downstream of the one analyzed. The treatment with intracytoplasmic sperm injection can result in vertical transmission of microdeletions of the AZF region and can also cause the expansion of a de novo mutation. This finding reinforces the necessity of an investigation of microdeletions of the Y chromosome in individuals who are candidates for assisted reproduction, as well as genetic counciling and follow-up.     

Fertilising capacity of human sperms: a simple predictive assay

Male subfertility is a growing reason for assisted reproduction. A limiting factor in male subfertility is asthenospermia. Motility is a cardinal indication of sperm vitality. Thus prognostic assays are aimed at quantitative determination of progression to assess the fertilising potential. However, a method permitting reliable prognosis of the fertilising capacity has yet to be developed. The assay presented here is the outcome of empirical data based on 590 IVF (in vitro fertilisation) trials. It is essentially a further exploitation of the Swim Up procedure, the selected sperms being maintained in culture under identical conditions employed in IVF. Semi-quantitative daily recordings of linear progression until complete extinction provided an index on vitality which is directly related to the fertilising potential. The findings indicated that a threshold of 50% linear motility after 24 hr culture was required to initiate fertilisation. The fertilising potential was guaranteed when at least 60% linear motility was observed at 24 hr, making the assay a predictive one. Its simplicity is an attractive feature.     

Affordable ART services in Africa: synthesis and adaptation of laboratory services

¹ Correspondence address. E-mail: carin.huyser{at}up.ac.za The aim of this paper is to provide information, opinions andsuggestions on affordable laboratory-orientated fertility screeningand treatment. Resource management to provide such servicesin developing countries, basic and advanced assisted reproductiveservices and assisted reproduction treatment (ART) of patientswith sexually transmitted infections are addressed. Alternativeviewpoints and parallel thinking should be encouraged to synthesizeand adapt first-world ART guidelines and recommendations intosafe and workable directives for developing regions. AffordableAfrican ART programmes, devoid of commercialism, can provideessential sexual health screening services en route to safefertility services for human immunodeficiency virus type-1 (HIV-1)serodiscordant couples (male HIV-positive), who wish to havetheir own biological child.     

Medical and socio-cultural aspects of infertility in the Middle East

³ Correspondence address: E-mail: giserour{at}thewayout.net The Middle East (ME), an area rich in history and traditionwith >300 million population, includes 18 heterogeneous countriesconcerning resources, income per capita, available healthcareservices, population density, growth rate, birth rate, totalfertility rate and life expectancy. There is a high prevalenceof infertility in the ME because of post-partum infection, unsafeabortion, iatrogenic tubal and pelvic infertility, tuberculosis,schistosomiasis and high incidence of male factor infertility.It is argued that in the ME, the solution to the problem ofinfertility is its prevention, and population control shouldtake precedence over infertility treatment. However, for a successfulfamily planning program and adoption of small family norms,couples should be reassured that they will be helped to achievepregnancy should they decide so. Prevention and treatment ofinfertility are of particular significance in ME because a womansocial status, her dignity and self-esteem are closely relatedto her ability to have children. Also there is gender sufferingof infertility in the ME. One of the stumbling blocks to acceptanceof assisted reproductive technology (ART) as a line of treatmentof infertility was the unacceptability to the main religiousgroups of the involvement of a third party in the act of procreation.Practices of ART in the ME have many common features and littledifferences. A mechanism had to be found to provide low-costART to the needy.     

Infertility-related reproductive health knowledge and help-seeking behaviour in African countries

¹Correspondence address. E-mail: silke.dyer{at}uct.ac.za Data from African countries indicate that men and women attributeinfertility to traditional beliefs about health and diseaseas well as to biomedical causes, although appropriate knowledgeof the latter is frequently lacking. Infertility is a dreadedcondition and as a result help-seeking is often intense andpersistent. Most of the help-seeking is undertaken by womenand both traditional and modern biomedical health services areaccessed. There are, however, many barriers to effective andaffordable biomedical infertility care, many of which are relatedto poor resources and lack of infrastructure, and as a resultthe need for infertility treatment is often unmet. Advancesin the quality of care require greater commitment to the problemof infertility in African countries, the provision of healtheducation as an integral part of infertility management, theintegration of infertility services into reproductive healthcare programmes and defining the role of assisted reproductivetechnologies in low resource settings. At the same time theimportance of traditional health services in infertility managementshould be recognized.     

Epigenetics, spermatogenesis and male infertility

Epigenetic modifications characterized by DNA methylation, histone modifications, and chromatin remodeling are important regulators in a number of biological processes, including spermatogenesis. Several genes in the testes are regulated through epigenetic mechanisms, indicating a direct influence of epigenetic mechanisms on the process of spermatogenesis. In the present article, we have provided a comprehensive review of the epigenetic processes in the testes, correlation of epigenetic aberrations with male infertility, impact of environmental factors on the epigenome and male fertility, and significance of epigenetic changes/aberrations in assisted reproduction. The literature review suggested a significant impact of epigenetic aberrations (epimutations) on spermatogenesis, and this could lead to male infertility. Epimutations (often hypermethylation) in several genes, namely MTHFR, PAX8, NTF3, SFN, HRAS, JHM2DA, IGF2, H19, RASGRF1, GTL2, PLAG1, D1RAS3, MEST, KCNQ1, LIT1, and SNRPN, have been reported in association with poor semen parameters or male infertility. Environmental toxins/drugs may affect fertility via epigenetic modifications. For example, 5-aza-2'-deoxycytidine, an anticancer agent, causes a decrease in global DNA methylation that leads to altered sperm morphology, decreased sperm motility, decreased fertilization capacity, and decreased embryo survival. Similarly, Endocrine disruptors, such as methoxychlor (an estrogenic pesticide) and vinclozolin (an anti-androgenic fungicide) have been found by experiments on animals to affect epigenetic modifications that may cause spermatogenic defects in subsequent generations. Assisted reproduction procedures that have been considered rather safe, are now being implicated in inducing epigenetic changes that could affect fertility in subsequent generations. Techniques such as intracytoplasmic sperm injection (ICSI) and round spermatid injection (ROSI) may increase the incidence of imprinting disorders and adversely affect embryonic development by using immature spermatozoa that may not have established proper imprints or global methylation. Epigenetic changes, in contrast to genetic aberrations, may be less deleterious because they are potentially reversible. Further research could identify certain drugs capable of reversing epigenetic changes.     

Sustained High Protein-tyrosine Phosphatase 1B Activity in the Sperm of Obese Males Impairs the Sperm Acrosome Reaction

Evidence of a causal link between male obesity and subfertility or infertility has been demonstrated previously. However, the mechanism underlying this link is incompletely understood. Here, we report that sustained high protein-tyrosine phosphatase 1B (PTP1B) activity in sperm of obese donors plays an essential role in coupling male obesity and subfertility or infertility. First, PTP1B level and activity were significantly higher in sperm from ob/ob mice than in wild-type littermates. High PTP1B level and activity in sperm was also observed in obese patients compared with non-obese donors. The enhanced sperm PTP1B level and activity in ob/ob mice and obese patients correlated with a defect of the sperm acrosome reaction (AR). Second, treating sperm from male ob/ob mice or obese men with a specific PTP1B inhibitor largely restored the sperm AR. Finally, blockade of sperm AR by enhanced PTP1B activity in male ob/ob mice or obese men was due to prolonged dephosphorylation of N-ethylmaleimide-sensitive factor by PTP1B, leading to the inability to reassemble the trans-SNARE complexes, which is a critical step in sperm acrosomal exocytosis. In summary, our study demonstrates for the first time that a sustained high PTP1B level or activity in the sperm of obese donors causes a defect of sperm AR and that PTP1B is a novel potential therapeutic target for male infertility treatment.     

Reproductive effort in invasive and non-invasive Rubus

We quantified the physiological costs and the total amount of resources allocated to reproduction in two closely related species of Rubus, one of which is invasive. These two species share several morphological and life-history characteristics and grow together in the Pacific Northwestern United States. Reproductive effort was manipulated in canes of both species by removing flower buds. The non-invasive species, R. ursinus, exhibited significantly greater water stress in the reproductive canes, as indicated by lower leaf water potential (O) and reduced stomatal conductance (g_s). This species also showed a reduction in leaf nitrogen concentration ([N]) associated with reproduction. Combined, these factors led to reduced photosynthesis (A) on a diurnal basis, lower water-use efficiency as inferred from '¹³C, and reduced photosynthetic capacity. All of these effects were more pronounced during the fruiting stage than in the flowering stage. The invasive species, R. discolor, showed no changes in water stress, [N], '¹³C, or A associated with reproduction. A model was used to estimate total gross photosynthesis (A_gross) for reproductive and non-reproductive canes of both species over cane lifetime. Reproduction was associated with a greater decline in A_gross for the non-invasive R. ursinus than for the invasive R. discolor. Although R. discolor allocated more resources directly to flowers and fruit than R. ursinus, the invasive species had significantly lower reproductive effort, or total amount of resources diverted from vegetative activity to reproduction, than the non-invasive species. By minimizing the reduction of photosynthesis associated with reproduction, this invasive species may be able to minimize the trade-offs commonly associated with reproduction.     

When should a mare go for assisted reproduction?

The use of assisted reproductive techniques (ART) has helped owners to produce offspring from valuable mares that were considered infertile using standard breeding techniques. Before referring a mare for an ART, the practitioner should be able to identify the underlying cause of subfertility of the mare. The objective of this review is to provide information regarding embryo transfer, oocyte transfer and intracytoplasmic sperm injection, the three most common ART used in equine practice. Knowing the complexity as well as the risks of these techniques, enables practitioners to refer a subfertile mare to the least complex and most appropriate and successful ART that can overcome specific causes of infertility.     

Autosomal mutations and human spermatogenic failure

Infertility, defined as the inability to conceive after 1 year of unprotected intercourse, is a healthcare problem that has a worldwide impact. Male factors are involved in at least half of these cases of infertility. Despite 33 years of assisted reproductive activities, a considerable number of cases (25–30%) remain idiopathic. This situation can be explained by a poor understanding of the basic mechanisms driving male and female gametogenesis. Compared to multi-organ pathologies, only a few non-syndromic genetic causes of human infertility have been described so far, despite the fact that it is estimated that some infertility cases could be explained by genetic causes and that over 200 infertile or subfertile genetic mouse models have been described. So far, very little has been discovered in the field of human male reproductive genetics. Consequently, genetic tests proposed to infertile couples are limited, although worldwide efforts devoted to the field of human genetics of infertility are expected to provide new genetic tests in the near future. We present the requirements for performing informative genetics studies in the field of infertility, the techniques used and the results obtained so far. This article is part of a Special Issue entitled: Molecular Genetics of Human Reproductive Failure.     

Sexual behavior and fertility of little mice

Male mice homozygous for the little gene mutation (lit) were originally reported to have a marked incidence of infertility. We have previously reported that these diminutive mice have normal spermatogenesis and testicular steroidogenesis. In the present study, one research goal was to determine if the reported infertility was due to a defect in male sexual behavior. Quantitative analyses of male sexual behavior of little mice and their normal siblings were completed. Natural breeding trials were also performed to reexamine the fertility of little male mice. The experimental results suggested that little mice require significantly longer times for the first mount, first intromission, and ejaculation. More importantly, the investigation provided evidence that the little mice were not infertile but that they were subfertile. The diminutive size of the little mice may be the primary etiological factor for the observed subfertility.     

Ethical issues of infertility treatment in developing countries

¹ Correspondence address. Tel/Fax: +32-16-620767; E-mail: guido.pennings{at}ugent.be The provision of infertility treatment in developing countriesis controversial. Reports over the last decades have inculcatedin people from Western countries the belief that overpopulationis the major problem of developing countries. This paper willanalyse the different arguments advanced for and against providinginfertility treatment to resource-poor countries. There aretwo arguments in favour: reproductive autonomy and the hugeburden of infertility in these countries. Pronatalism, whichreigns in almost all developing countries, is to a great extentresponsible for the devastating effects of infertility. Thefive arguments against the application of infertility treatmentare overpopulation, prioritization of limited resources, preventionrather than cure, justice and equal access and risk of abuse.The importance of a person's reproductive autonomy demands thatefforts should be made to enable people to determine how manychildren to have. This is equally true in developing countries.However, given the enormous difficulties of resource-poor countriesto provide even the most basic goods, the contribution by societyshould be directed mostly at prevention and should depend ona strong cost reduction for assisted reproductive technology.     

Increased frequency of CFTR gene mutations identified in Indian infertile men with non-CBAVD obstructive azoospermia and spermatogenic failure

High incidence of mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene is associated with congenital bilateral absence of the vas deferens (CBAVD) and is considered as the genital form of cystic fibrosis (CF). The CFTR gene may also be involved in the etiology of male infertility in cases other than CBAVD. The present study was conducted to identify the spectrum and frequency of CFTR gene mutations in infertile Indian males with non-CBAVD obstructive azoospermia (n = 60) and spermatogenic failure (n = 150). Conspicuously higher frequency of heterozygote F508del mutation was detected in infertile males with non-CBAVD obstructive azoospermia (11.6%) and spermatogenic failure (7.3%). Homozygous IVS(8)-5T allele frequency was also significantly higher in both groups in comparison to those in normal healthy individuals. Two mutations in exon 25 viz., R1358I and K1351R were identified as novel mutations in patients with non-CBAVD obstructive azoospermia. Mutation R1358I was predicted as probably damaging CFTR mutation. This is the first report from the Indian population, emphasizing increased frequency of CFTR gene mutations in male infertility other than CBAVD. Thus, it is suggested that screening of CFTR gene mutations may be required in infertile Indian males with other forms of infertility apart from CBAVD and willing for assisted reproduction technology.     

Causes of secondary sexual differences in plants — Evidence from extreme leaf dimorphism in Leucadendron (Proteaceae)

In extreme cases leaves in male plants of the dioecious genus Leucadendron (Proteaceae) are up to an order of magnitude smaller than female leaves. This secondary sexual dimorphism (SSD) in leaf size has previously been suggested to be due to intra-male sexual selection, leading to an increase in male allocation to reproduction in dimorphic species. After critically evaluating previous data provided to support this hypothesis, I suggest on both theoretical grounds and on re-analysis that this argument is unlikely and unsupported. Leaf size dimorphism could theoretically evolve directly due to disruptive ecological selection between genders, leading to niche dimorphism either within or between habitats. I test this ecological causation hypothesis by providing data on specific leaf area (sla) and water use efficiency (δ ¹³C) of leaves from males and females of several Leucadendron species. Results confirm the expectation of minimal gender differences. I argue that leaf dimorphism is a consequence of selection on flower size and architecture.     

Pathogens that cause infertility of bulls or transmission via semen

The purpose of this paper is to review scientific evidence regarding pathogens that cause infertility of bulls or that could be transmitted via bovine semen. Although several pathogens can cause male infertility and potentially be transmitted via semen, adhering to disease control recommendations provided by Certified Semen Services (CSS) and the World Organization for Animal Health (OIE) can prevent infectious male infertility and ensure that the risk of pathogen transmission via semen is negligible. Regarding bulls to be used for natural breeding, quarantine prior to herd introduction and appropriate diagnostic testing during quarantine will commonly prevent introduction of pathogens that adversely affect reproduction.