Tests de Selection des Spermatozoïdes avant Assistance Médicale à la Procréation

There are many procedures to select spermatozoa but those procedures are often specific of each andrology centre. This review that is not an exhaustive review of the numerous methods published is rather devoided to focus on the main techniques, the sperm washing, the filtration methods («SpermPrep»), the swim-up, and the discontinuous density gradients with colloïdal solutions («Percoll», PureSperm” and «Isolate») including iodinated organic molecule as ‘OptiPrep». The first part presents a synthesis of literature in order to bring out standardized procedures, and a summary of the advantages and disadvantages of each step of these procedures. Some specific anomalies of sperm need particular treatments. There is a consensus on the validated techniques to prepare semen samples in the presence of antisperm antibodies, leukocytes, bacterial contaminants, and also semen samples exhibiting hyperviscosity or in case of retrograde ejaculation. Cryopreserved sperm preparation is also described with references to experimental results in freezing-thawing human spermatozoa. The second part proposes guidelines for the therapeutic choice of the appropriate assisted reproductive technologies (i.e.: intra uterine artificial insemination, in vitro fertilization or intracytoplasmic sperm injection) according to semen parameters, and to the outcome of sperm selection and survival.     

Cryoconservation de spermatozo?des avant vasectomie: utilité et paradoxes à travers l’activité des CECOS

Sperm banking when consulting for vasectomy can appear as a paradox. Around 12% of the men who decide to have a vasectomy consult in a CECOS for sperm cryopreservation before surgery. Among these men, less than 5% will use their frozen sperm. In about 10 years, artificial insemination has been quite abandoned and in vitro fertilization is now widely used by these patients when they desire a child in a new couple. It is thus time to ask wether cryopreservation of sperm is still to be proposed before vasectomy.     

Perspectives de l’analyse de la mobilité des spermatozoïdes

Several studies have shown a good correlation between sperm motility and fertility though the microscopic evaluation of the percentage of motile sperm is highly subjective by nature. Therefore in the last decade, various objectives methods have been proposed to overcome this problem. Two types of methods were developed: The methods based on the analysis of images obtained by microphotography, microcinematography and microvideography and the global, undirect methods based on physical principles. Several systems based on video and image analysis (Computer Aided Sperm Analysis, CASA) have been developed and are used in numerous laboratories of reproductive biology. CASA technology offers the possibility to analyse some characteristics of sperm motion which are related to the fertilization potential and to develop new parameters related to some important aspects of sperm behavior such as hyperactivation. However, there is a large amount of interactions between the operator and the CASA machine. CASA instruments are not “ready-to-use” robots: the reliability of CASA depends largely on the expertise and training of the user and the application of standardized procedures and quality control schemes. By contrast, there is only minimal interaction between the operator and the Sperm Quality Anlyser which is a new device measuring and index of sperm motility highly correlated to the concentration of progressively motile sperm. The device uses light passed through a small sample of semen introduced in a capillary tube to detect variations in optical density that result from moving particles. The reproducibility of the measurements is excellent, the device is easy to use and this is a potentially useful tool for field-work studies. Further investigations of this device in the managment of male infertility is warranted. Finally, both types of objectives approaches are complementary to the conventional analysis of sperm motility and they will not replace it. Standardized procedures have been proposed by the World Health Organization for the subjective evaluation of sperm motility. Such procedures are very useful to reduce significantly the intra- and interlaboratory variations but internal and external quality controls schemes indicate that they are not sufficient to achieve acceptable levels of variation and regular quality controls followed by the definition and the application of corrective procedures are required.     

Traçabilité en assistance médicale à la procréation

An ART traceability procedure provides a written record of the controls performed to ensure precise allocation of gametes or embryos during insemination or IVF techniques. This procedure requires analysis of the various steps associated with a risk of error and optimization of safety systems. The author provides a basis for further thought on this subject.     

Evaluation de l’apport de la méthode d’observation des spermatozoïdes à fort grossissement en ICSI

Due to the growing interest in the method of high-magnification sperm observation and selection proposed for the specific indication of ICSI failure, the authors evaluated the technique in unselected ICSI. The aim of this study was to evaluate the relevance of Motile Sperm Organelle Morphology Examination (MSOME) compared with usual selection performed in ICSI. In a series of conventionally selected sperm for ICSI, the number with an abnormal appearance on high magnification was determined and the predictive value of this parameter on ICSI outcome was assessed. The study included 25 successive unselected ICSI attempts in the IVF Laboratory of Poissy Hospital (France). ICSI were performed according to usual protocols used in the laboratory. Twenty five motile spermatozoa of the migrated fraction, still available after ICSI, and “injectable— according to conventional morphology assessment in ICSI (“normal” or “as normal as possible” with magnification of ×200–400) were assessed by MSOME (higher than ×4500) and classified according to criteria adapted from Bartoov’s work and taking into account David’s sperm morphology classification. We compared the results of MSOME and ICSI results. In this small series of ICSI with diverse indications, we found very high frequencies of abnormalities (more than 70%), particularly nuclear vacuoles. No predictive value of the morphology of sperm assessed with high magnification (including vacuoles) was found for fertilization rate, embryo quality and ICSI outcome. In contrast with previous reports, pregnancies were obtained with very abnormal sperms. In this series of unselected ICSI, nuclear vacuoles do not seem to have a pejorative impact on pregnancy outcome. This study raises several perspectives. It would be interesting to understand the “anatomical” basis for vacuoles observed with MSOME and their meaning. The question of the phenotype-genotype relation, i.e. the possible correlation between sperm morphology and genetic content could be investigated. Finally, a prospective analysis should be performed in clearly defined indications to validate the potential applications of the method for high-magnification sperm observation and selection.     

Stratégies biologiques à l’heure de l’Assistance Médicale à la Procréation chez le blessé médullaire

Most men with spinal cord injury (SCI) men have fertility problems caused by anejaculation and decreased fertility of the ejaculate. There are two main causes for the impaired reproductive potential in SCI men: ejaculatory dysfunction and poor quality semen. However, current treatment techniques allow a large number of SCI males to achieve ejaculation (rectal electro-stimulation, penile vibrator stimulation). Firstly, masturbation and/or penile vibrator are used at home allowing the couple to perform insemination themselves. The semen of men with spinal cord injuries has commonly been characterised by small volume, abnormal count (low or high), decreased sperm mobility, increased formation of reactive oxygen species, sperm autoimmunity, necrospermia. This impairment is thought to be due to insufficient drainage, genitourinary infections and raised scrotal temperature. Testicular biopsy reveals varying degrees of tubule degeneration and decreased spermatogenetic activity. Semen could be used for various assisted reproductive technologies such as intrauterine insemination,in vitro fertilisation (IVF) and microinsemination (ICSI). The literature reports pregnancy rates by intrauterine insemination of about 15 to 20% per couple. Clinical pregnancy rates after IVF and ICSI techniques are 30% per cycle and these results are comparable to the clinical pregnancy rates when these techniques are performed for female infertility. However, semen must be frozen as soon as possible after the injury and the patient must be informed about the various available assisted reproductive technologies.     

Facteurs influençant la détectabilité de l’activité myocardique en caméra à semi-conducteurs

Body weight was already shown to be a major factor for the detectability of myocardial activity with conventional cameras and thus, for the optimization of injected activities and or recording times. This study was aimed to identify the factors affecting this detectability on a new solid-state camera showing a high sensitivity of detection (D.SPECT camera Cyclopharma^®).Patients and methodsThe study population involved 101 patients, who underwent myocardial SPECT with Sestamibi on a one-day stress/rest protocol. Each conventional acquisition (DST-XL General Electric Healthcare^® USA) was immediately followed by a D-SPECT^® one. Myocardial activity was determined on each acquisition in counts per seconds and expressed in fraction of the injected activity for assessing the detectability of myocardial activity.ResultsMyocardial activity was on average 12 ± 3 fold higher when recorded with the D-SPECT^® camera than with the DST-XL^® one. Body weight and especially body mass index (BMI) were the most significant correlates of the detectability of myocardial activity for both cameras. According to these correlations, an increase in the BMI from 25 to 35 kg/m was associated with a 47% decrease in the detectability of myocardial activity at stress with the DST-XL^® and of only 30% with the D-SPECT.ConclusionThe detectability of myocardial activity provided by the D.SPECT camera is dramatically higher than that documented for conventional cameras of the DST-XL^® type and is affected by the increase in body weight and in body mass index but at a lower rate than for the Anger camera.     

Intérêt de l’analyse de la morphologie fine et de la qualité nucléaire des spermatozoïdes dans le cadre des techniques d’AMP

Routine semen examination does not identify minor malformations of the sperm nucleus and chromatin architectural defects, which may be associated with ART outcome and cannot be detected by the embryologist even at 1000x magnification. Recent publications have demonstrated the advantages, compared to routine analysis, of a new method of real-time detailed morphological evaluation of motile spermatozoa: motile sperm organellar morphology examination (MSOME). MSOME is performed with an inverted light microscope equipped with high-power differential interference contrast optics enhanced by digital imaging to achieve a magnification of 10000x. To be considered morphologically normal, a sperm nucleus must have both a normal shape and a normal chromatin content. The aim of the present study was to combine MSOME and sperm DNA fragmentation characteristics to assess reproductive outcome. The study population consisted of the male partners of 52 couples referred for conventional IVF or split cycles (half IVF-half ICSI cycles) and exhibiting normal routine sperm parameters. Spermatozoa were analysed by examining the fine nuclear morphology and DNA integrity using the sperm chromatin dispersion test (SCD test), based on the principle that the deproteinized nuclei of spermatozoa with nonfragmented DNA show extended halos of DNA dispersion that are either absent or only minimally present in sperm nuclei with fragmented DNA. Fertilization rates were significantly lower in the group showing less than 8% of normal spermatozoa according to MSOME criteria, but early embryo development was not affected. Fine sperm morphology correlated with DNA fragmentation rate. These results demonstrate that the assessment of sperm nuclear normality by MSOME analysis and SCD test improves characterization of the semen sample and should be evaluated as a tool for allocating patients to specific assisted reproduction treatments.     

De l’origine des anthropoïdes à l’émergence de la famille humaine

The human lineage has a very ancient origin, as most of the mammals. Its oldest representatives, anthropoid primates, have been described from Asia some 45 million years ago. During this long evolutionary story, two critical stages have appeared as especially important, their beginning in Asia and the emergence of hominids in Africa, some seven million years ago. These two stages are discussed hereby with new data relative to their Asian origins and their dispersal into Africa between 45 and 40 million years ago. Following this dispersal event, these primates evolved in Africa and gave rise to the early hominids. These appeared around seven million years ago and have three distinct representatives. Among them, Toumaï appears as the oldest and the closest to our ancestry, a point that is evidenced here.     

ComorbiditéS associées au trouble déficit de l’attention avec hyperactivité : Données épidémiologiques et implications thérapeutiques

ADHD has become one of the most frequent cause of referrals for children’ behaviour disorders. ADHD is a prevalent psychiatric condition affecting 5% to 9% of school-age children with regards to DSM-IV R diagnostic criteria. In addition, according to the results of different epidemiological studies, patients with ADHD very often experience comorbid conditions in 50% to 90% of the cases. The most frequent comorbidities include externalised disorders, (oppositional defiant disorder, conduct disorders), learning disorders, internalised disorders (anxiety, depression) and tics (chronic motor tics, Tourette’s syndrome). Given their negative impact on the outcome of ADHD in terms of affective and social functioning, and of social and school adaptation, these comorbid conditions should be carefully and systematically searched, even without any actual complaint. Although management of comorbid psychiatric conditions is never simple nor straightforward, therapeutic option should be considered taking into consideration both management of ADHD and specificity of these comorbidities.     

Méthode simple d’évaluation de l’activité volumique d’effluents radioactifs au moyen d’une caméra à scintillations

The aim of this technical note is to evaluate an easy method to measure ^99mTc samples with an activity of 1000, 100 and 10 Bq/L. This study is performed with a gamma-camera detector in two departments of Nuclear medicine in Avignon and in Nîmes. We develop a procedure to measure ^99mTc radioactive waste at the two hospitals output in accordance with the DGS/DHOS no. 2001/323 circular requests of the Ministry for Employment and Solidarity.     

La procréation médicalement assistée avec spermatozoïdes immatures: Impératifs cliniques et techniques chirurgicales

The authors report their experience with the use of spermatids in TESE programs where mature spermatozoa could not be isolated from testicular biopsies. The details of the indications for spermatid insemination, the technicity of the procedure and the results are exposed.     

Groupe de prise en charge clinico-biologique en Assitance Médicale à la Procréation (AMP) des Patients à Risque Viral

La prise en charge en Assitance Médicale à la Procréation concerne de plus en plus de couples à risque viral. Cette prise en charge est aujourd’hui le seul moyen efficace de réduire une transmission viral mère-enfant ou entre partenaires séro-différents. Elle met en jeu une importante activité pluridisciplinaire où travail en commun et concertation entre les différentes équipes médicales clinico-biologiques sont les deux points clefs.     

Intérêt de l’étude de l’oxydation de l’ADN des spermatozoïdes par marquage de la 8-oxo-guanine en cytométrie en flux chez l’homme infertile

Oxidative stress in semen is essentially due to excessive production of oxygen-reactive species (ORS) essentially derived from leukocytes. DNA oxidation is due to the direct action of ORS which produce several adducts the most extensively studied of which is 8-oxo-guanine. Integrity of DNA is essential for the fertility of sperm and is an important subject of research for scientists and clinicians all over the world. Although evaluation of the global integrity of sperm DNA has considerably developed over recent years, few tests are available to document oxidative DNA damage. This study was designed to review the various tests of sperm DNA integrity commonly used in the literature and to present the results of our study on DNA oxidation with 8-oxo-guanine labelling by flow cytometry in infertile men. This study was based on 15 semen samples that were submitted to sperm analysis according to WHO guidelines, with determination of the leukocyte concentration by a cytochemical method revealing peroxidase in cytoplasmic granulations. The DNA oxidation study was performed with 8-oxo-guanine labelling by flow cytometry. Linear regression analysis showed a strong correlation between DNA oxidation and leukocyte count in the semen (p = 0.006, r = 0.7). A leukocyte cut-off of 250,000/ml of semen was associated with a significant increase of DNA oxidation (p = 0.03). 8-oxo-guanine can therefore be considered to be a biological marker of the direct action of oxidative stress on sperm DNA which appears to be susceptible to relatively low levels of ORS produced by leukocytes present at concentrations well below the limit of leukospermia defined by WHO.