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Biomedical science is rapidly developing in terms of more transparency, openness and reproducibility of scientific publications. This is even more important for all studies that are based on results from basic semen examination. Recently two concordant documents have been published: the 6th edition of the WHO Laboratory Manual for the Examination and Processing of Human Semen, and the International Standard ISO 23162:2021. With these tools, we propose that authors should be instructed to follow these laboratory methods in order to publish studies in peer-reviewed journals, preferable by using a checklist as suggested in an Appendix to this article.

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RESUME La conservation des ovocytes s’effectue par vitrification, en France depuis 2011, et seules y sont autorisees les conservations destinees a la preservation de la fertilite, donc les indications medicales. Cette technique s'adresse surtout aux femmes atteintes de pathologies malignes, dont un traitement gonadotoxique, par chimiotherapie ou radiotherapie risque de degrader la fonction ovarienne. Elle concerne pareillement les femmes menacees d'insuffisance ovarienne prematuree. A ces indications se sont ajoutees depuis quelques annees des demandes dites « non medicales », en fait de palliation de l'infertilite liee a l'avancee en âge. Sans perspective previsible de grossesse, surtout faute de stabilite de couple, des femmes sensibles a la baisse de la fertilite avec l'âge, souhaitent faire prelever et conserver leurs ovocytes afin d’y avoir eventuellement recours plus tard, par FIV, si elles eprouvaient alors des difficultes a concevoir. Les chances de grossesse dependent essentiellement de l'âge auquel les ovocytes ont ete recueillis, au mieux avant 35 ans. Les risques et les avantages possibles de ces demarches, sont discutes. Non autorisees en France elles sont effectuees par les femmes francaises dans les centres etrangers. Cependant la Loi de Bioethique du 07 juillet 2011 a etendu aux femmes majeures n'ayant jamais procree la possibilite de participer au don d’ovocytes. En contrepartie ces femmes peuvent conserver pour elles-memes, si leur nombre le permet, une partie des ovocytes recueillis. Ainsi est ouverte la voie legale de la conservation des ovocytes pour palliation de l'infertilite liee a l'âge.

The performance of the molecular tool using CRISPR-Cas9, which makes it possible to induce targeted modifications of the DNA, has found numerous applications in research and open promising prospects in human clinic. CRISPR-Cas9 has been widely used to generate transgenic animals after targeted modification of the genome at the zygotic stage. It was also tested on human embryos on an experimental basis. Although there are potential medical indications that may justify a targeted modification of the embryo or germ cell genome, the uncertainties regarding the efficacy and safety of the method do not allow us to consider implementing such germline gene therapy in the short-term. However, it is necessary to weigh the scientific and ethical issues involved in this approach.

RESUME Les interventions ayant pour but de modifier le genome de la descendance sont proscrites depuis 1994 en France mais le developpement de methodes comme CRISPR-Cas9 conduit a s’interroger sur leurs utilisations potentielles sur les cellules germinales et l’embryon humains. La seule indication medicale acceptable serait d’eviter la transmission d’une pathologie genique a l’enfant mais les conditions, notamment celles relatives a l’efficacite et a l’innocuite de ces methodes, ne sont pas actuellement reunies pour envisager leur utilisation clinique. De plus il existe d’autres moyens permettant aux couples concernes de realiser leur projet parental. Les questions ethiques suscitees par ces technologies incitent a recommander l’ouverture d’une reflexion pluridisciplinaire qui devrait etre menee dans le cadre d’un debat plus large portant sur l’ensemble des interventions medicales realisees lors de de l’assistance medicale a la procreation, pouvant avoir des consequences sur le genome des enfants a naitre et eventuellement sur celui des generations suivantes. En revanche, les recherches, y compris sur les cellules germinales et l’embryon humains, devraient pouvoir etre menees quand elles sont scientifique- ment et medicalement justifiees.

Study question Can a standardized assessment of abnormal human sperm morphology provide additional useful information by identifying men with more severe disturbances in different types of abnormalities? Summary answer Definition-based categorization of sperm head, midpiece and tail defects has shown how differently these abnormalities are distributed in fertile men and other groups of men, thus providing high and low thresholds, a starting point for diagnosis or research purposes. What is known already Several recent studies have reported indisputable genetic origins for various sperm defects. A few studies have also identified associations between environmental factors and low percentages of morphologically normal spermatozoa. Nevertheless, with the exception of rare situations in which the vast majority of spermatozoa have specific, easily characterized defects, such as 'globozoospermia', little attention has been paid to the description and precise quantification of human sperm abnormalities. The lack of standardization in the phenotyping of sperm morphological defects by conventional microscopy is a limiting factor for diagnosis and for intra- or inter-observer or centre consistency in studies investigating the causal factors and possible functional consequences of the abnormalities detected. There are currently no baseline data for abnormalities of sperm morphology based on a standardized classification, in the general population, among fertile or other groups of men. Study design, size, duration This study is based on detailed sperm abnormality datasets acquired by a standardized classification method, from several groups of men, over the same 5-year period. Participants/materials, setting, methods We studied cross-sectional data from fertile men (n = 926), male partners from infertile couples (n = 1747) and testicular cancer patients (n = 239). We used a standardized classification to analyse Shorr-stained slides, taking into account all the abnormalities encountered. Main results and the role of chance Most sperm defects were significantly more frequent in infertile than in fertile men, with 20-30% of infertile men having frequencies of abnormalities above the 95th percentile in fertile men for 9 out of the 15 categories of abnormalities. Interestingly, several head abnormalities were significantly more frequent in patients with testicular cancer than in infertile men, highlighting the particular impact of this condition on sperm morphogenesis. We used the 95th percentile in fertile men as the lower threshold and the 99th percentile in infertile men as an extreme upper threshold, for the classification of morphological abnormality frequencies into three levels: low, intermediate and high. The assessment of several semen samples, with or without a genetic background, for abnormal sperm morphology, based on the percentage of normal spermatozoa, a teratozoospermia index, and the detailed profile of abnormalities categorized according to the three levels proposed, has highlighted the value of detailed phenotyping for diagnosis and research purposes. Limitations, reasons for caution The thresholds proposed for the various categories of sperm abnormality should be considered relative rather than absolute, owing to the known sampling error related to the limited number of spermatozoa assessed per sample, or when studying the general population or populations from regions other than Western Europe. The standardized assessment of abnormal sperm morphology requires time and experience. We therefore suggest that this assessment is carried out during a first andrological check-up or for epidemiological or research studies, rather than in the routine management of infertile couples for assisted reproductive technologies. Wider implications of the findings The study design used for the fertile group of men was similar to that previously used for the WHO reference values, providing a rationale for considering the 95th percentile in fertile men as the level below which abnormalities may be considered to occur at a frequency representing random background variations of a normal spermiogenesis process. The crude frequencies obtained, and the three levels of abnormality frequency proposed for each standardized category of sperm defect, provide baseline data useful for diagnosis and a starting point for future studies aiming to identify associations with genetic or environmental factors. Study funding/competing interests Part of this study was supported by contract BMH4-CT96-0314 from the European Union. The authors have no competing interests to declare.

STUDY QUESTION Are antiretroviral therapies associated with semen alterations in HIV-infected men? SUMMARY ANSWER Antiretroviral regimens that included the non-nucleosidic reverse transcriptase inhibitor efavirenz were associated with a significant impairment of sperm motility, whereas regimens without efavirenz were not associated with significant semen changes. WHAT IS KNOWN ALREADY Semen alterations including decreased ejaculate volume and sperm motility have been reported in HIV-infected men. The hypothesis ascribing reduced sperm motility to damages induced in sperm mitochondria by nucleosidic (or nucleotidic) reverse transcriptase inhibitors (NRTIs) has not been confirmed in HIV-infected patients and the effects of antiretroviral treatments on semen parameters remain unclear. STUDY DESIGN, SIZE, DURATION This case-control study compared semen characteristics across 378 HIV-1 infected patients receiving different antiretroviral regimens or never treated by antiretroviral drugs, in whom an initial semen analysis was done between 2001 and 2007. PARTICIPANTS/MATERIALS, SETTING, METHODS The patients were partners from serodiscordant couples requesting medical assistance to procreate safely. Their status with regard to antiretroviral therapy at the time of semen analysis was categorized as follows: 1/ never treated patients (n = 66); 2/ patients receiving NRTIs only (n = 49); 3/ patients receiving a NRTIs + protease inhibitor (PI) regimen (n = 144); 4/ patients receiving a NRTIs + non-nucleosidic reverse transcriptase inhibitor (NNRTI) regimen (n = 119). Semen parameters were assessed through standard semen analysis. Additional analyses included measurement of sperm motion parameters using computer-assisted semen analysis, seminal bacteriological analysis, seminal biochemical markers and testosterone plasmatic levels. All analyses were performed in the Cochin academic hospital. The data were analyzed through multivariate analysis. MAIN RESULTS AND THE ROLE OF CHANCE Sperm motility was the only semen parameter which significantly varied according to treatment status. The median percentage of rapid spermatozoa was 5% in the group of patients receiving a regimen including efavirenz versus 20% in the other groups (P < 0.0001). Accordingly, sperm velocity was reduced by about 30% in this group (P < 0.0001). The role of chance was minimized by the strict definition and the size of the study population, which included a large enough group of never treated patients, the controlled conditions of semen collection and analysis, the multivariate analysis, the specificity and the high significance level of the observed differences. LIMITATIONS, REASONS FOR CAUTION The design of the study did not allow demonstrating a causal link between exposure to efavirenz and sperm motility. WIDER IMPLICATIONS OF THE FINDINGS As efavirenz is widely used in current antiretroviral therapy, these findings may concern many HIV-infected men wishing to have children. This justifies further assessment of the consequences on fertility of the exposure to efavirenz. Moreover, the possibility of common cellular impacts underlying adverse effects of efavirenz in sperm cells and neurons deserved investigation. STUDY FUNDING/COMPETING INTERESTS No external funding was used for this study. None of the authors has any conflict of interest to declare.

RESUME Apres avoir fait le recit des evenements qui ont precede la premiere transplantation uterine ayant abouti a la naissance d’un enfant vivant et bien portant et fait un etat des lieux, les auteurs exposent la legislation francaise actuelle sur la greffe d’organes. Puis ils abordent les aspects cliniques, insistant sur la complexite de l’acte chirurgical, le dilemme du choix entre donneuse en etat de mort cerebrale ou decedee et donneuse vivante et les indications chez la receveuse. Ils decrivent ensuite le traitement immunosuppresseur avant et pendant la grossesse, les complications plus ou moins graves qui peuvent en emailler le cours et la surveillance particulierement attentive qu’elles necessitent, mais aussi les doutes sur l’opportunite de l’allaitement maternel. Ils s’interrogent sur l’avenir de l’enfant a moyen et long terme, son developpement psychomoteur et celui de son systeme immunitaire. Ils retracent les nombreuses et delicates questions ethiques que pose la transplantation uterine, qu’il s’agisse des particularites de la greffe d’uterus, qui n’est pas vitale mais permet de donner la vie, du choix entre transplantation avec donneuse en etat de mort cerebrale ou donneuse vivante, de la penurie d’organes a greffer, des risques courus par la receveuse, du devenir physique et psychologique de l’enfant, enfin du choix entre transplantation uterine et gestation pour autrui et de l’eventualite de derives. En fait, la transplantation uterine est une chirurgie encore au stade experimental et seuls l’avenir et le recueil exhaustif de toutes les donnees la concernant permettront de s’assurer de son bien-fonde.

The performance of the molecular tool using CRISPR-Cas9, which makes it possible to induce targeted modifications of the DNA, has found numerous applications in research and open promising prospects in human clinic. CRISPR-Cas9 has been widely used to generate transgenic animals after targeted modification of the genome at the zygotic stage. It was also tested on human embryos on an experimental basis. Although there are potential medical indications that may justify a targeted modification of the embryo or germ cell genome, the uncertainties regarding the efficacy and safety of the method do not allow us to consider implementing such germline gene therapy in the short-term. However, it is necessary to weigh the scientific and ethical issues involved in this approach.

RESUME En France comme dans d’autres pays, de plus en plus de femmes vivant seules ou en couple homosexuel souhaitent devenir meres. Pour mener a bien leur projet parental, elles peuvent consulter un medecin en France et souvent se rendent a l’etranger pour beneficier d’une Assistance Medicale a la Procreation avec les spermatozoides d’un donneur (AMPD). L’AMPD devrait-elle leur etre accessible en France ? Il n’appartient pas au medecin de le decider. L’AMPD realisee dans ces circonstances n’a pas d’indication ni de contre-indication medicale. Ce mode de procreation suscite de nombreuses questions concernant le developpement et le bien-etre des enfants. Les resultats des etudes menees dans d’autres pays sont souvent rassurants mais les conditions de leur obtention ne permettent pas de conclure et de nombreuses zones d’ombre persistent. Il serait donc necessaire de poursuivre l’effort de recherche recemment initie en France dans le domaine. Si l’acces a l’AMPD des femmes sans partenaire masculin etait legalise, il devrait etre accompagne de mesures pour assurer sa mise en oeuvre selon les principes ethiques de benevolat et d’anonymat s’appliquant a tout don d’element du corps humain. Ces mesures devraient aussi permettre une prise en charge medicale efficace des actes et de leurs consequences pour favoriser au mieux le bien-etre des enfants et de leurs meres.

Le Cecos (Centre d’Etude et de Conservation des Œufs et du Sperme) de l’Hopital Cochin a ouvert en 1999 un programme pour l’insemination artificielle avec donneur (IAD) de la compagne d’un homme d’origine transsexuelle (transman). Ce projet avait suscite des reticences, discutees dans un comite d’ethique?: interrogation sur la sante mentale du transman, sa capacite a etre pere, sur la possibilite d’un developpement normal chez les enfants, en particulier sur la construction de leur identite sexuee. Un suivi des enfants fut propose aux parents pour savoir si les objections faites etaient pertinentes, auquel cas il faudrait interrompre le programme, ou non pertinentes, auquel cas on serait fonde a poursuivre le programme et en proposer la generalisation aux autres Cecos. Ce suivi a ete accepte des parents, qui ont ete revus avec leurs enfants a la fin de la premiere annee de l’enfant, puis tous les deux ans, dans le cadre d’une consultation. De 2000 a 2010, 42 enfants (28 garcons et 14 filles) sont nes de 29 couples et, a la fin de 2010, 24 enfants avaient ete suivis, les autres devant etre vus l’annee suivante. Ce suivi montre qu’il ne s’agit pas de couples homosexuels?; ces couples sont stables?; le pere se conduit en pere competent?; mais ce sont des peres sans penis qui s’identifient aux valeurs masculines et paternelles de leur culture. Quant aux enfants, ils vont bien, se developpent normalement, ont l’air d’etre heureux de vivre et se sentent des garcons ou des filles en accord avec leur sexe biologique. Il avait ete recommande aux parents de dire la verite sur l’IAD et le transsexualisme du pere. Pour l’IAD, il n’y a guere eu de problemes. Pour le transsexualisme, malgre leur desir d’en parler a l’enfant, les peres ne trouvaient pas les mots pour le dire?; un livret ecrit et dessine par l’un des peres les a aides, montrant qu’il s’agissait de parler simplement de leur enfance telle qu’ils l’avaient vecue, ce que les peres ont souvent du mal a faire spontanement. Il n’y a pas lieu de regretter d’avoir permis a des transmen d’acceder a la paternite. Il sera interessant de voir comment leurs enfants vivront leur adolescence.

In France as in other countries, more and more single women and lesbian couples wish to become mothers. To carry through their parenting project they may consult a physician in France and often go abroad in order to get Assisted Reproductive Technologies with donor sperm (ARTD). Should ARTD be available to those women in France? The physician has not to take the decision. In such situations ARTD has no medical indication or contraindication. This assisted procreation raises many questions on children development and well-being. The results of studies made in other countries are often reassuring but their methodologies do not allow any conclusion to be drawn and grey areas persist. Therefore it should be necessary to develop a research effort in the field as it recently started in France. Would ARTD access to women without a male partner be legalized, the law should respect the ethical principles of non-payment and anonymity associated with donation of all body components. In any case, it should also allow an efficient medical care to be performed to ensure under the best conditions the well-being of the children and their mothers.

RESUME En 2011, l’Academie nationale de medecine a ete saisie de la delicate question de l’assistance medicale a la procreation (AMP) en prison, question deja ancienne puisque remontant a 1977, mais recurrente en raison de sa complexite, de sa gravite et des interferences qu’elle implique avec les structures judiciaires et penitentiaires. Apres avoir rappele, d’une part les modifications de la loi penitentiaire francaise et le contenu de la recente loi bioethique, d’autre part le contexte carceral si particulier de l’AMP en France et dans d’autres pays, le rapport aborde les nombreuses questions suscitees par de telles demandes. Il s’interroge d’abord sur le sens donne actuellement a l’incarceration et sur les possibilites d’avoir des rapports sexuels et de procreer en prison. Il constate que la realisation d’une AMP dans ces conditions doit etre placee sur le meme plan que le traitement de toute autre pathologie et discute la compatibilite de l’AMP avec les deux termes de la loi bioethique : infertilite medicalement prouvee et persistance d’une communaute de vie. Le rapport envisage ensuite les interets en jeu et souligne l’importance d’apprecier a sa juste valeur le projet parental du couple intentionnel et de le confronter a l’interet de l’enfant, son bien-etre et son avenir. Puis le rapport envisage les difficultes de la mise en œuvre de l’AMP en prison, les multiples contraintes materielles qu’elle suscite ainsi que les aspects financiers. Il detaille les facteurs devant intervenir dans la decision : le parcours de l’incarceration, l’âge des protagonistes, la situation respective des parents intentionnels, les antecedents personnels et familiaux, la qualite du projet parental et les relations existant entre les futurs parents, leur etat de sante physique et psychique, leur environnement familial, enfin la cause de leur incarceration et leurs antecedents judiciaires.

RESUME Concernes par l’incidence croissante des cancers du sein, de la prostate et du testicule dans les pays industrialises, y compris en Europe, nous avons fait une revue de la litterature scientifique et des rapports francais sur les effets potentiellement carcinogenes des perturbateurs endocriniens (PEs) presents dans l’environnement et l’alimentation. Nous expliquons pourquoi il est tres difficile d’obtenir une preuve epidemiologique d’un effet cancerigene des PEs dans l’espece humaine, ce qui explique en partie les polemiques interminables sur cette question. Cependant les resultats d’une serie d’etudes independantes sont suffisants pour evoquer fortement un effet cancerigene de ces substances, notamment en cas de cancers hormono-dependants. Ces resultats proviennent d’etudes experimentales faites chez les rongeurs et des mesures de niveaux d’exposition dans le sang et les urines chez l’humain. La lecon de l’effet cancerigene transgenerationnel qui a ete observe apres traitement de femmes enceintes par le diethylstilbestrol et les resultats obtenus apres exposition experimentale de rongeurs au bisphenol A suggerent que les femmes enceintes et/ou allaitantes et les jeunes enfants doivent etre proteges en priorite. Alors qu’il a deja ete decide de reduire la presence de certains PEs comme les pesticides, les dioxines et les PCBs dans l’environnement, nous proposons de nouvelles mesures de precaution ciblees sur le bisphenol A et les phtalates qui sont notamment utilises dans le conditionnement des aliments. Cependant, avant d’interdire l’usage du bisphenol A dans les emballages alimentaires, il serait necessaire que les chercheurs industriels et academiques agissent de maniere concertee pour mettre au point des substituts au bisphenol A faisant preuve d’une meilleure innocuite.

BACKGROUND: Through oocyte donation (OD), women with Turner syndrome (TS) may achieve motherhood. However, this popu- lation has a high prevalence of cardiac malformations and carry a risk for aortic dissection that is increased by pregnancy. Until recently, the necessity for a specialized cardiac evaluation before pregnancy was underestimated as was the need for follow-up through adulthood. The aim of this study was to evaluate the follow-up (mainly cardiovascular) of women with TS requesting OD. METHODS: Disease monitoring since diagnosis and prior cardiac evaluations conducted out of our centre were assessed in 25 women with TS who requested OD. New cardiac evaluations using echocardiography and magnetic resonance imaging were performed by our specialized cardiologist in I8 of these patients. RESULTS: We observed that the medical follow-up of women with TS was often deficient throughout adulthood. Most of the prior cardiac evaluations performed by cardiologists not accustomed to women with TS, either before (n = 8) or when starting OD (n = 12), were con- sidered normal. However, when revaluated by a cardiologist who is familiar with TS, seven women were diagnosed with a bicuspid aortic valve and thus excluded from OD. In addition, when appropriate screening was conducted by our referent cardiologist before OD no cardiac complication was observed during pregnancy or delivery. CONCLUSIONS: Careful follow-up, including cardiac evaluation, should be recommended for women diagnosed with TS, before and after puberty. Moreover, assessment of cardiovascular parameters by a cardiologist familiar with TS should be routinely repeated before under- taking OD.

RESUME A la parution du rapport d’information de la mission parlementaire consacre a la revision des lois de bioethique, l’Academie nationale de medecine a juge utile d’entreprendre une etude critique de ce rapport et, en particulier des 95 propositions qui sont avancees, afin de formuler a leur egard les points d’accord, des remarques, des propositions de redaction plus appropriee sur certains points, ou des reserves. Chacun des chapitres de la Loi a ete examine: assistance medicale a la procreation, gestation pour le compte d’autrui, diagnostics ante natals, examen des caracteristiques genetiques, recherche sur les embryons humains et les cellules embryonnaires, brevetabilite des elements et produits du corps humain, don d’organes et de tissus, le respect et l’identite du corps de la personne decedee, neurosciences, nanotechnologies et convergence des technologies. Les 95 propositions formulees par la Mission Parlementaire sont ainsi passees en revue dans le souci d’apporter au legislateur, avec la plus large adhesion possible, la contribution de notre Institution.

Objective To collect follow-up data for infertile men with Y microdeletion. Design Retrospective, observational survey. Setting Multicenter IVF units associated with genetics laboratories. Patient(s) Sixty-three patients with Y microdeletion. Intervention(s) Karyotype analysis, Y microdeletion screening, and assisted reproductive technology. Main Outcome Measures Medical history, karyotype, nature of the AZF deletion, semen parameters, testis biopsy results, choice of assisted reproductive technology, and results of intracytoplasmic sperm injection (ICSI). Results Abnormal karyotypes were found in 8 men (12.7%), who were azoospermic except 1. Of these 8 men, 5 presented a combined AZFb+c deletion, and 3 had a deletion in AZFc only. Most men (39 of 63) were azoospermic, 3 were cryptoazoospermic, and 19 had extreme oligozoospermia (sperm concentration ≤1.10 6 /mL). Sperm concentration above 1.10 6 /mL was found for 2 men (3.1%). A testis biopsy was performed in 27 azoospermic men, resulting in positive sperm extraction in 6 cases. To date, 42 ICSI cycles with either testicular (n = 5) or ejaculated spermatozoa (n = 37) have been carried out in 23 couples with male partners with AZFc deletion. Eighteen clinical pregnancies were obtained, leading to the birth of 14 babies. Donor insemination had been chosen by 28 couples, leading to the birth of 9 children. Conclusion Karyotype analysis should be systematically performed in Y microdeleted men. Intracytoplasmic sperm injection can be offered to half of AZFc-deleted patients, providing real opportunities to have a child.

Children conceived through assisted reproductive technologies (ART) now account for a noteworthy proportion (-2.4%) of births in France. Considerable attention is being paid to the outcome of ART pregnancies. The vast majority of these children are apparently normal. However, they are at an increased risk of minor birth defects, low birth weight, and rare imprinting disorders such as Beckwith-Wiedemann syndrome (BWS), Angelman syndrome (AS) and Silver Russel syndrome (SRS). Animal models are important for investigating the possible role of each step of ART (ovarian stimulation, gamete manipulation, in vitro fertilization, embryo culture and embryo transfer) in epigenetic reprogramming This review discusses these issues in the context of epigenetic and developmental abnormalities observed in animals following ART More research is needed on ART-induced errors, focusing not only on genomic imprinting but also on non-imprinted loci, which may help explain some of the more subtle longer-term health effects emerging from studies with animal models.

L’esperance de vie accrue des patients seropositifs pour le virus humain de l’immunodeficience (VIH) depuis l’avenement des tritherapies en 1996 leur permet desormais d’envisager des projets de vie a long terme dont celui d’exprimer leur desir d’enfants naturels via, malgre leur fertilite, une assistance medicalisee pour reduire le risque d’une contamination de la femme et du fœtus. Les difficultes psychologiques eventuellement liees a cette situation n’avaient pas encore ete decrites. L’objectif de cette etude etait de decrire la psychologie et le projet d’enfant de ces couples et le developpement des bebes nes au cours de leurs premiers mois de vie. Les couples ont ete evalues immediatement apres l’inclusion dans le protocole a l’aide d’un entretien clinique et standardise (SCID for DSM-IV) et avec des autoquestionnaires (Questionnaire tridimensionnel de la personnalite et Echelle d’harmonie de la dyade). Un deuxieme entretien, clinique et standardise, associe a l’echelle Edinburg Postnatal Depression Scale pour les femmes a eu lieu au 6e mois de la grossesse. La 3e evaluation a eu lieu au cours des deux premieres annees du post-partum avec ces memes instruments. Le developpement des enfants a ete evalue au cours de leurs deux premieres annees a l’aide d’un examen clinique et par les tests de Bayley et de Brunet-Lezine, ainsi qu’avec l’echelle Alarme Detresse Bebe et le questionnaire de Denver complete par les deux parents. 176 couples ont ete inclus, 165 ont beneficie d’une AMP, 150 d’une evaluation psychologique. 52 % des couples ont obtenu un enfant avant la fin du protocole. Aucune seroconversion n’a ete constatee. Les couples ne presentaient pas davantage de troubles psychiatriques que la population generale. 102 bebes sont nes, 45 ont ete evalues. A l’exception de deux jumeaux, tous les bebes presentaient un developpement psychomoteur normal. Le succes de cette etude demontre l’importance de cette assistance pour les couples serodifferents dans leur projet d’enfant. Il s’agit de la premiere etude a demontrer le bon developpement des bebes ainsi concus.

Summary In this paper are presented the results obtained by the light scattering method of a systematic study of different parameters characterizing spermatozoa motility in a series of human sperms, one hour and five hours after ejaculation. These results are relative to the concentration of spermatozoa, the percentage and the number of motile spermatozoa, the characteristic velocity and the velocity distributions. All these parameters are discussed one by one, the advantages (objectivity, small quantity of not diluted sperm, short time needed for measurements) and the limits of the light scattering method being pointed out. A comparison is made with results obtained by conventional methods. The fineness of this study shows that this method can be used with success in fundamental research as well as in a routine functional sperm study, allowing an objective and precise analysis of motility in a given population as a complex factor. Zusammenfassung Eine systematische Studie der verschiedenen fur die Motilitat der Spermatozoen kennzeichnenden und durch die Lichtstreuungsmethode gerechneten Parameter wurde in dieser Arbeit zusammengefast. Die Untersuchungen umfassen eine Serie eine Stunde und funf Stunden nach der Ejakulation erhaltener menschlicher Spermatozoen. Die Spermatozoenkonzentration, der Prozentsatz an beweglichen Spermatozoen, die Anzahl der letzteren, die charakteristische Geschwindigkeit und die Geschwindigkeitsverteilung wurden jeweils angegeben. Jeder Parameter wurde einzeln im Zusammenhang mit den Vorteilen der Methode (d.h. ihrer Objektivitat, der Anwendung von relativ kleinen Mengen unverdunnten Spermas, den schnell gerechneten Werten) sowie deren Grenzen diskutiert. Anschliesend wurden die Befunde mit den durch konventionelle Methoden erhaltenen Ergebnissen verglichen. Wegen ihrer Prazision kann diese Methode erfolgreich im Gebiet der Grundlagen-Forschung sowie im Rahmen einer funktionellen Routinespermatozoenstudie angewendet werden, indem sie eine objektive und genaue Analyse der Motilitat der Spermatozoen als komplexen Faktors ermoglicht. Resumen Se presentan los resultados obtenidos por el Metodo de dispersion optica de un estudio sistematico de diferentes parametros que caracterizan la motilidad espermatica en una serie de espermas humanos, 1 y 5 horas despues de la eyaculacion. Estos resultados son relativos a la concentracion de espermatozoides moviles, caracteristica y distribucion de la velocidad. Se discuten todos estos parametros uno por uno, la ventaja (objetividad, pequena cantidad de esperma no diluido, rapidez de la medicion) y se apuntan los limites del metodo de dispersion optica. Se comparan los resultados con los obtenidos por metodos convencionales. La perfeccion de este estudio muestra que este metodo se puede utilizar con exito en investigacion fundamental aski como en el estudio funcional rutinario del espermatozoide, permitiendo un analisis preciso y objetivo de la movilidad en una poblacion dada.

When natural conception is impossible and the underlying problem cannot be treated, medical intervention can reproduce the steps necessary for fertilization and early embryo development. The first known medical action in the field of human reproduction took place at the end of the 18th century, in the form of artificial insemination with the husband's semen, thus dissociating sexual intercourse from procreation. A further upheaval occurred at the end of the 19th century, with the use of donor sperm, separating the notions of genetic descent and parenthood. In the second half of the 20th century, medically assisted procreation saw two major technological advances, namely gamete freezing and in vitro fertilization (IVF). The first child conceived with frozen-thawed sperm was born in 1953, and the first IVF baby in 1978. Fertilization by intracytoplasmic sperm injection (ICSI), first developed in 1992, can overcome many causes of male infertility. The convergence of reproductive biology and genetics has now opened up the possibility of screening for chromosome and gene defects in the embryo, prior to implantation. Thus, assisted reproductive technologies (ART) not only serve as a substitute for natural conception but can also avoid the birth of a disabled child While new technologies continue to extend the available options for infertile couples, they also have the potential to help single women and homosexual couples to have children. These practices are currently only accepted in certain countries. Overall, these new medical technologies have contributed to changing our conception of human reproduction, opening up new paradigms of parenthood and raising new challenges for society.

The aim of this study was to investigate whether a relationship exists between the presence of low numbers of leukocytes in normal ovulatory cervical mucus and sperm quality and lipid content after migration. The percentages of live, motile and morphologically normal spermatozoa, movement parameters assessed by computer-aided sperm analysis (CASA), and ionophore-induced acrosome reaction measured by flow cytometry were determined before and after migration. High-performance liquid chromatography with ultraviolet detection was used to measure the sperm lipid content, including the various diacyl subspecies. The number of leukocytes found in solubilized mucus samples was counted using a haemocytometric method. Overall, the presence of leukocytes in the cervical mucus samples did not significantly influence sperm motility and morphology, sperm kinematic parameters, or the sperm content in sphingomyelin or cholesterol. In contrast, after migration, the decrease in various sperm diacyls and the level of induced acrosome reaction was significantly less pronounced in mucus samples containingor=10(4) leukocytes than in mucus samples with no or rare leukocytes whereas the level of induced acrosome reaction was higher. The present data suggest that the low level of leukocytes found in normal ovulatory cervical mucus could influence the process of sperm lipid remodelling/capacitation.

Testicular germ cell tumours (TGCTs) are the most common malignancies in Caucasian young men and their incidence has increased over the past decades. However, a non-invasive test allowing an early diagnosis of TGCT often proves inaccurate. We have previously shown that two Cancer-Testis Antigens (CTA), namely MAGE-A4 and NY-ESO-1, were expressed by TGCT. As exfoliation of carcinoma in situ (CIS) cells or tumour germ cells from testis into seminal fluid can occur, here we studied the expression of the 2 CTA in semen smears of patients with testicular cancer in comparison with healthy men. Using semen smears from healthy controls (n = 65) and patients diagnosed for testicular tumour (n = 57) and immunological staining, we observed expression of MAGE-A4 and NY-ESO-1 proteins in seminal fluid exfoliated cells. We found a highly statistically significant difference in the ratios of stained cells to the total number of round cells between testicular cancer patients and healthy controls. Multivariable analysis, including sperm parameters and immunostaining on sperm smears, shows the improvement. This technique can provide towards testicular cancer diagnosis when it is included in the current testing regime. However, the fact that expression of these markers was not restricted to foetal germ cells led to detection in the semen of a number of healthy subjects. Although the detection of these CTA could be useful to characterize the sub-type of individual TGCTs better, we stress here that the false positive rate precludes the exclusive employment of these CTA for the early detection of testicular neoplasia.

Resume Des l’introduction des techniques d’Assistance medicale a la procreation (AMP), de nombreuses etudes ont ete menees pour evaluer leur impact sur la sante des enfants ainsi concus. La notion de risque epigenetique a ete evoquee et un lien entre AMP et pathologies associees a des anomalies de l’empreinte parentale a ete suggere : en particulier, dans le cas des syndromes de Beckwith-Wiedemann (BWS), d’Angelman (AS) et de Silver-Russell (SRS). Le cycle des marques epigenetiques (ecrasement dans les lignees germinales primordiales, etablissement de l’empreinte au cours des gametogeneses, lecture et maintien de l’empreinte dans les cellules somatiques) accompagne toutes les phases de la reproduction, la gametogenese, la maturation gametique, la fecondation et le developpement embryonnaire precoce. L’objectif de cette revue est de recenser toutes les informations issues de l’experimentation animale pouvant contribuer a une meilleure connaissance des perturbations de l’empreinte parentale, en liaison avec les differentes techniques utilisees en AMP et fournir des informations utiles dans la perspective d’ameliorer la prise en charge des couples engages dans un projet parental.

Points essentiels En France, l’arrete ministeriel du 10 mai 2001 autorise les personnes infectees par le virus de l’immunodeficience humaine (VIH) a recourir a l’assistance medicale a la procreation (AMP) dans le but de reduire le risque de transmission entre les partenaires ou de traiter une infertilite du couple. La personne infectee doit avoir un taux sanguin de lymphocytes CD4 > 200/mm 3 et une charge virale stable (absence d’augmentation de plus de 0,5 log 10 copies/mm 3 ) sur 2 prelevements dans les 6 mois precedant l’AMP. Les co-infections (VHC, VHB) doivent faire l’objet d’une evaluation par un specialiste. Lorsque l’homme est infecte, seul le recours a l’AMP permet de concevoir tout en maintenant des rapports proteges au sein du couple. L’AMP est realisee avec des spermatozoides prepares et valides par une recherche negative d’ARN du VIH. La methode d’AMP est determinee par le bilan de fertilite du couple et par la quantite de virus dans le liquide seminal. Le traitement antiretroviral n’est pas requis pour l’AMP mais peut s’averer necessaire lorsque la charge virale seminale est elevee. Lorsque la femme est infectee, le couple doit etre informe des risques de transmission du VIH a l’enfant et des risques de toxicite des traitements antiretroviraux pour la mere comme pour le fœtus. Ces risques doivent guider l’optimisation du traitement antiretroviral, qui n’est pas systematique lors de l’AMP mais le sera au dernier trimestre de la grossesse. Le projet d’enfant doit etre pris en charge en proposant des auto-inseminations et le recours a l’AMP propose rapidement car la fonction ovarienne semble precocement alteree chez les femmes infectees. Plusieurs milliers de couples en Europe ont beneficie de l’AMP sans contamination rapportee a ce jour. Environ la moitie de ces couples peut esperer avoir un enfant, mais pres d’un couple sur 3 ne s’engage pas dans l’AMP apres consultation.

The case of a successful pregnancy is reported here, following intracytoplasmic sperm injection which resulted in a cohort of embryos showing accelerated cleavage kinetics. Pregnancy was not achieved with fresh embryos transferred at the 4-cell stage but was achieved with cryopreserved accelerated embryos. After an ovarian stimulation cycle, 20 mature oocytes were observed. Early cleavage was observed in 19 of these, 26 h post-injection, and 17 were beyond the 4-cell stage 42 h after injection. No pregnancy was obtained after the transfer of two fresh embryos at the 4-cell stage without fragments. Following the transfer of two frozen accelerated embryos, a twin pregnancy was obtained, which resulted in the delivery of two healthy babies. This case report demonstrates that classical scoring systems may need to be adapted to reflect the cleavage kinetics of the whole embryo cohort.

RESUME Depuis quelques dizaines d’annees, la lutte contre l’infertilite a considerablement progresse grâce au developpement de techniques biomedicales maitrisant la maturation des gametes, la fecondation et le developpement de l’embryon avant l’implantation. Les nombreuses questions ethiques, sociales et medicales suscitees par l’assistance medicale a la procreation (AMP) ont conduit le legislateur a definir en 1994 les conditions d’acces aux actes d’AMP et les modalites de leur realisation. Le dispositif a ete complete recemment par la creation de l’Agence de la Biomedecine. En 2005, 123 000 cycles de traitement ont ete realises en France toutes techniques confondues. Ils ont conduit a la naissance de 19 026 enfants. Les taux de grossesse clinique ont varie de 11 a 24,4 % par tentative selon les techniques. Il s’agissait de grossesses multiples dans 11 a 21 % des cas. Aujourd’hui, l’objectif prioritaire est d’ameliorer la qualite des resultats, c’est a dire de maitriser autant que possible les risques et les contraintes de l’AMP tout en maintenant les meilleures chances d’obtenir une grossesse et une naissance. Une meilleure caracterisation des embryons jointe a une meilleure evaluation de la fertilite de la femme permet de reduire le nombre d’embryons transferes et donc le risque de grossesse multiple sans diminuer les chances de procreation. Les principaux developpements envisageables a court et moyen terme concernent l’utilisation de traitements hormonaux moins agressifs pour stimuler la fonction ovarienne, une meilleure selection des gametes fonctionnels, des conditions de fecondation et de culture embryonnaire plus adaptees. Par ailleurs la mise au point de micro-methodes capables d’evaluer les competences metaboliques des embryon, et leur aptitude a se developper normalement, devrait conduire a une amelioration significative des resultats.

Fertilization in mammals requires an initial interaction of sperm with the oocyte envelope, the zona pellucida (ZP), before it reaches the oocyte. ZP is a highly glycosylated structure, composed of three (mouse) or four (rabbit, boar, bovine, humans...) glycoproteins. The presence of ZP around the oocyte does not allow heterospecific fertilization. This barrier is principally due to the presence of species-specific glycosylations on ZP proteins. Sperm bind ZP by means of membrane receptors which recognize carbohydrate moieties on ZP glycoproteins according to a well-precised sequential process. Upon initial attachment, spermatozoa bind ZP3/ZP4 which induces the sperm acrosome exocytosis followed by a secondary binding of acrosome reacted spermatozoa to ZP2 and by ZP penetration. The sperm receptors are adhesive proteins or integral plasma membrane proteins linked to intraspermatic signalling pathways activating the acrosome reaction. Over the last twenty years, numerous studies have been carried out to identify sperm receptors to ZP in several species, but the data in humans are still incomplete. Work initiated in our research group has identified several proteins interacting with recombinant human ZP2, ZP3 and ZP4, among which are glycolytic enzymes. These enzymes are involved in the gamete interaction by means of their affinity to sugars and not by their catalytic properties. From a clinical point of view, an observed lack or weak expression of some sperm receptors to ZP3 in cases of idiopathic infertility associated with in vitro fertilization failure suggests that knowing the molecular mechanism driving the gamete recognition can be important at the diagnostic level. Furthermore, it has been shown that proteins that mediate gamete recognition diverge rapidly, as a result of positive darwinian selection. A sexual conflict can drive co-evolution of reproductive molecules in both sexes resulting in reproductive isolation and species emergence.

Testicular cancer (TC) risk factors remain largely unknown, except for personal history of cryptorchidism and familial history of TC. We conducted a hospital-based case-control study on familial, environmental and occupational conditions in which we compared 229 cases and 800 controls. TC was correlated with cryptorchidism (OR = 3.02; CI: 1.90-4.79), a history of cryptorchidism in relatives (OR = 2.85; CI: 1.70-4.79), and TC (OR = 9.58; CI: 4.01-22.88], prostate cancer (OR = 1.80; CI: 1.08-3.02) and breast cancer (OR = 1.77; CI: 1.20-2.60) in relatives. Living in a rural area or having regular gardening activity (growing fruit or vegetables) was associated with an increased risk of TC (OR = 1.63; CI: 1.16-2.29; OR = 1.84; CI: 1.23-2.75). Regarding occupation, we found a relationship with employment in metal trimming (OR = 1.96; CI: 1.00-3.86), chemical manufacture (OR = 1.88; CI: 1.14-3.10), industrial production of glue (OR = 2.21; CI: 1.15-4.25), and welding (OR = 2.84; CI: 1.51-5.35). In a multivariate model, only a history of cryptorchidism in the men, cryptorchidism in relatives, TC, and breast cancer remained significant. Our findings contribute further evidence to a pattern of TC risk factors, which include the significant weight of personal reproductive history and also of testicular and breast cancer in relatives. By including in a multivariate model variables linked to environmental and occupational exposure and related to familial cancer history, neither living in a rural area nor any occupational exposure appeared to be a potential environmental TC risk factor.

Study question Can a tool be developed for authors, reviewers and editors of the ESHRE Journals to improve the quality of published studies which rely on semen analysis data? Summary answer A basic checklist for authors, reviewers and editors has been developed and is presented. What is known already Laboratory work which includes semen analysis is burdened by a lack of standardization. This has significant negative effects on the quality of scientific and epidemiological studies, potential misclassification of patients and the potential to impair clinical treatments/diagnoses that rely on accurate semen quality information. Robust methods are available to reduce laboratory error in semen analysis, inducing adherence to World Health Organization techniques, participation in an external quality control scheme and appropriate training of laboratory personnel. However, journals have not had appropriate systems to assess if these methods have been used. Study design, size, duration After discussion at a series of Associate Editor Meetings of the ESHRE Journals the authors of the present text were asked to propose a tool for authors, reviewers and editors of the ESHRE Journals to ensure a high quality assessment of submitted manuscripts which rely on semen analysis data, including a detailed verification of the relevance and the quality of the methods used. Participants/materials, setting, methods N/A. Main results and the role of chance A basic checklist for authors, reviewers and editors is presented. The checklist contains key points which should be considered by authors when designing studies and which provides essential information for when the submitted manuscript is evaluated. For published articles the answers in the checklist are suitable to be available as supplementary data, which will also reduce the space necessary for technical details in the printed article. Limitations, reasons for caution Guidelines such as these should not be used uncritically. It is therefore important that submitting authors, in situations where their study does not comply with the basic requirements for semen analysis, not only explain all methodological deviations but also declare the level of uncertainty in their analyses and how it complies with, or might confound, the aims of the study. Wider implications of the findings The fundamental importance of appropriate and robust methodology to facilitate advances in scientific understanding and patient management and treatment, is now accepted as being paramount. Use of the semen analysis checklist should be part of this process, and when completed and signed by the corresponding author at the time of submitting a manuscript should result in greater transparency, and ultimately uniformity. It is hoped that this initiative will pave the way for wider adoption of the methodology/reporting by other biomedical, epidemiological and scientific journals, and ultimately become the standard of practice for papers reporting semen analysis results obtained in laboratory and clinical andrology. Systems to assist referees, authors and editors to present high quality findings should have a significant impact on the field of reproductive medicine. Study funding/competing interests No funding was obtained for this work. The authors have no competing interests in relation to the present publication and checklist. Trial registration number N/A.

Some transsexual persons wish to have their gametes frozen before gender transition, in order to preserve their fertility. This measure should be carried out, in strict compliance with the law, in case of orchidectomy, oophorectomy or hysterectomy However, as hormonal treatments do not irreversibly alter gonadal function, the reproductive capacity of trans-sexual persons can be maintained by avoiding surgical sterilization. There is therefore no obvious medical indication for cryopreserving gametes or germinal tissue in the absence of surgical sterilization. Moreover, the use of such cryopreserved gametes would, in principle, be considered mainly by a same-sex couple, something that French law currently prohibits. Regardless of these legal aspects, the issues surrounding the use of cryopreserved gametes, and its consequences, must not be ignored. If transsexual persons who are already parents may find ways of managing the change in both their personal and parental identity, the use of gametes stored prior to gender transition raises issues of identity whose consequences are difficult to assess, especially for the future child. Cryopreservation of gametes or germinal tissue cannot be undertaken without first considering whether their potential use is in keeping with what is, at present, medically and legally possible. In any case, it is up the physician to decide, on a case by case basis, whether or not to implement cryopreservation, taking into account the situation of the persons who request the procedure and their plans for parenthood.

Resume Outre les contraintes physiques importantes, les techniques d'assistance medicale a la procreation s'accompagnent d'une lourde charge emotionnelle et psychologique, a la fois pour la femme et pour l'homme. Les couples concernes sont generalement infertiles, mais il existe des situations particulieres ou les personnes sont fertiles, comme celles des couples dont l'homme est seropositif, et qui recourent a ces techniques pour eviter les risques de contamination lies aux rapports sexuels non proteges. Les couples sont generalement soudes et motives, mais les difficultes sont parfois importantes, notamment l'attente de la grossesse et la deception liee a l'echec d'un cycle. Les reactions les plus frequentes sont le stress, l'anxiete et les depressions, mais les difficultes sont parfois telles que certains couples renoncent apres une seule tentative. Globalement, les parents infertiles ne presentent pas davantage de difficultes a s'occuper de leurs enfants que les parents fertiles. Les enfants, malgre un risque accru d'anomalies genetiques, presentent dans l'immense majorite des cas un developpement normal. Cependant, certains enfants presentent des difficultes psychologiques particulieres qui peuvent justifier d'un suivi specialise.

Resume Les progres des traitements anti-retroviraux permettent maintenant aux personnes infectees par le VIH d’aspirer a fonder une famille. La reglementation en France les autorise depuis 2001 a recourir a l’assistance medicale a la procreation (AMP) dans le but de reduire le risque de transmission entre les partenaires ou de traiter une infertilite du couple. Le controle de l’infection a VIH et de co-infections a VHC ou VHB est un prealable indispensable, ainsi que l’accompagnement psychologique du couple. Lorsque l’homme est infecte, seul le recours a l’AMP permet de concevoir tout en maintenant des rapports proteges au sein du couple. Le sperme est concentre et le VIH recherche dans la fraction finale des spermatozoides. Les echantillons congeles valides sont alors utilisables en AMP. Lorsque la femme est infectee, le couple doit etre informe des risques de transmission du VIH a l’enfant et du traitement optimise pour les reduire. De simples auto-inseminations peuvent etre proposees au couple, apres un bilan d’infertilite si la femme a plus de 38 ans. Le projet d’enfant doit etre pris en charge rapidement et le recours a l’AMP propose en cas d’echec des methodes non medicalisees, car la fonction ovarienne semble precocement diminuee chez les femmes infectees. Si celle-ci est normale, les chances de succes sont celles de l’AMP habituellement. Les grossesses gemellaires doivent etre evitees car elles favorisent la transmission materno-fœtale du VIH. Plusieurs milliers de couples en Europe ont beneficie de l’AMP sans qu’aucune contamination n’ait ete rapportee. Environ la moitie des couples pris en charge peuvent esperer avoir un enfant.

Background: Genomic imprinting refers to an epigenetic marking resulting in monoallelic gene expression and has a critical role in fetal development. Various imprinting diseases have recently been reported in humans and animals born after the use of assisted reproductive technology (ART). All the epimutations implicated involve a loss of methylation of the maternal allele (demethylation of KvDMR1/ KCNQ1OT1 in Beckwith–Wiedemann syndrome (BWS), demethylation of SNRPN in Angelman syndrome and demethylation of DMR2/ IGF2R in large offspring syndrome), suggesting that ART impairs the acquisition or maintenance of methylation marks on maternal imprinted genes. However, it is unknown whether this epigenetic imprinting error is random or restricted to a specific imprinted domain. Aim: To analyse the methylation status of various imprinted genes ( IGF2R gene at 6q26, PEG1/MEST at 7q32, KCNQ1OT1 and H19 at 11p15.5, and SNRPN at 15q11–13) in 40 patients with BWS showing a loss of methylation at KCNQ1OT1 (11 patients with BWS born after the use of ART and 29 patients with BWS conceived naturally). Results: 3 of the 11 (27%) patients conceived using ART and 7 of the 29 (24%) patients conceived normally displayed an abnormal methylation at a locus other than KCNQ1OT1 . Conclusions: Some patients with BWS show abnormal methylation at loci other than the 11p15 region, and the involvement of other loci is not restricted to patients with BWS born after ART was used. Moreover, the mosaic distribution of epimutations suggests that imprinting is lost after fertilisation owing to a failure to maintain methylation marks during pre-implantation development.

BACKGROUND: Sterility in men with cystic fibrosis (CF) raises the question of the use of assisted reproductive techniques (ART). A multidisciplinary network including adult CF centre and reproductive medicine units was set up to answer requests for ART. METHODS: This retrospective study included 25 men with CF between 1994 and 2004. Clinical status, semen analysis, CF mutations analysis and artificial insemination by donor (AID) or ICSI after surgical retrieval of sperm were monitored. RESULTS: All CF men had azoospermia. Two chose AID first (one delivery); 23 chose ICSI. Sperm were surgically retrieved in 21 cases, and ICSI was performed in 19. Pregnancies occurred in 12 of these 19 couples (63%) (two ectopic pregnancies, two spontaneous abortions, one termination of pregnancy for polymalformed twins and 11 single deliveries in nine couples). Two couples tried AID after ICSI failed; one had twins. Another adopted a child, and two had a spontaneous pregnancy. After a follow-up of 4.1±2.3 years, two patients died, two underwent lung transplantation and 21 remained stable. CONCLUSION: ART can help men with CF to become a father, but their health status and short survival need careful counselling and multidisciplinary medical care.

La prise en charge de l’infertilite a debute il y a plus de deux siecles par la realisation d’inseminations artificielles, technique interferant peu sur le processus de reproduction puisque la fecondation se deroulein vivo. Les annees 80 ont vu l’emergence de la fecondationin vitro (FIV) necessitant une stimulation hormonale soutenue, une fecondation exvivo et une culture embryonnaire. Enfin, l’arrivee de l’ICSI (Intra Cytoplasmic Sperm Injection) a revolutionne la prise en charge des couples en permettant de creer un embryon par simple microinjection d’un spermatozoide, meme anormal ou immature, dans un ovocyte. Rapidement, la question de l’impact de ces techniques sur la sante des enfants ainsi concus a ete posee. Des etudes recentes ont suggere un lien entre aide medicale a la procreation et developpement de pathologies d’empreinte. En effet, l’empreinte parentale etablie specifiquement sur chacun des gametes et transmise a l’embryon peut etre sensible a des facteurs externes. La manipulation et la culturein vitro des gametes et embryons, inherentes a la realisation de la FIV/ICSI, pourraient avoir des consequences sur l’etablissement et/ou la conservation de l’empreinte. La connaissance des mecanismes d’acquisition et de maintien de l’empreinte et l’analyse de leur deregulation sont necessaires pour evaluer ce risque potentiel.

Objective: To compare the prevalence of hepatitis C (HCV) RNA in semen from men infected with HCV and those coinfected with HIV-1 /HCV and to study the origin of HCV shed in semen. Design: Two prospective studies (HC EP09 and BINECO) included 120 HCV-positive men, 82 coinfected with HIV-1; all had positive HCV RNA detection in blood. Methods: Paired blood and semen samples were collected for HCV RNA detection and quantification in seminal plasma and in blood serum; repeated semen samples were obtained for 45 men. HCV RNA was sought in spermatozoa and non-sperm cells. Phylogenetic analysis of the HVR-1 region of HCV compared the quasispecies in blood serum and seminal plasma of two men. Results: HCV RNA was more frequently found in the semen of men coinfected with HIV-1 (37.8%) than in those with only HCV infection (18.4%) (P = 0.033). HCV RNA detection in semen was intermittent and was positive in at least one semen sample of 42.8% of HIV-1/HCV-coinfected men who provided repeated samples. Men with HCV-positive semen had significantly higher HCV load in blood than men with HCV-negative semen (P = 0.038). Phylogenetic comparison of HCV quasispecies in blood and in semen showed no evidence of HCV replication in genital leukocytes; however, a phenetic structure was observed between compartments (P < 0.001). Conclusions: HCV particles in semen originate from passive passage from blood, with preferential transfer of some variants. Nearly half of HIV-1/HCV-coinfected men may intermittently harbour HCV in their semen. Recommendations of protected sex for HIV-infected individuals should be reinforced.