Human Reproductive Medicine Research Articles

The mutual benefits of research in wild animal species and human-assisted reproduction.

Studying the reproductive biology of wild animal species produces knowledge beneficial to their management and conservation. However, wild species also share intriguing similarities in reproductive biology with humans, thereby offering alternative models for better understanding the etiology of infertility and developing innovative treatments. The purpose of this review is to raise awareness in different scientific communities about intriguing connections between wild animals and humans regarding infertility syndromes or improvement of fertility preservation. The objectives are to (1) highlight commonalities between wild species and human fertility, (2) demonstrate that research in wild species-assisted reproductive technologies can greatly enhance success in human reproductive medicine, and (3) recognize that human fertility preservation is highly inspiring and relevant to wild species conservation. In addition to having similar biological traits in some wild species and humans, the fact of sharing the same natural environment and the common needs for more options in fertility preservation are strong incentives to build more bridges that will eventually benefit both animal conservation and human reproductive medicine.

Scrambled eggs: Proteomic portraits and novel biomarkers of egg quality in zebrafish (Danio rerio).

Egg quality is a complex biological trait and a major determinant of reproductive fitness in all animals. This study delivered the first proteomic portraits of egg quality in zebrafish, a leading biomedical model for early development. Egg batches of good and poor quality, evidenced by embryo survival for 24 h, were sampled immediately after spawning and used to create pooled or replicated sample sets whose protein extracts were subjected to different levels of fractionation before liquid chromatography and tandem mass spectrometry. Obtained spectra were searched against a zebrafish proteome database and detected proteins were annotated, categorized and quantified based on normalized spectral counts. Manually curated and automated enrichment analyses revealed poor quality eggs to be deficient of proteins involved in protein synthesis and energy and lipid metabolism, and of some vitellogenin products and lectins, and to have a surfeit of proteins involved in endo-lysosomal activities, autophagy, and apoptosis, and of some oncogene products, lectins and egg envelope proteins. Results of pathway and network analyses suggest that this aberrant proteomic profile results from failure of oocytes giving rise to poor quality eggs to properly transit through final maturation, and implicated Wnt signaling in the etiology of this defect. Quantitative comparisons of abundant proteins in good versus poor quality eggs revealed 17 candidate egg quality markers. Thus, the zebrafish egg proteome is clearly linked to embryo developmental potential, a phenomenon that begs further investigation to elucidate the root causes of poor egg quality, presently a serious and intractable problem in livestock and human reproductive medicine.

Activation of Nrf2 might reduce oxidative stress in human granulosa cells

Nuclear factor-E2-related factor 2 (Nrf2)/Kelch-like ECH-associated protein 1 (Keap1)-antioxidant response element (ARE) signaling pathway is one of the most important defense mechanisms against oxidative stress (OS). It is well documented that equilibration status of OS plays fundamental roles in human reproductive medicine, and the physiological role of Nrf2 in ovarian granulosa cells (GCs) has not been determined yet. Herein we aimed to study the function of Nrf2 in GCs. Human ovarian tissues were subjected to immunohistochemistry to localize Nrf2 and Keap1 and we detected the expression of Nrf2 and Keap1 in the human GCs. Human luteinized GCs were isolated and cultured, and hydrogen peroxide (H2O2) or Dimethylfumarates (DMF), an activator of Nrf2, were added to GCs to analyze the relationship between Nrf2 and antioxidants by quantitative RT-PCR. The mRNA levels of Nrf2, catalase, superoxide dismutase 1 (SOD1), and 8-Oxoguanine DNA glycosylase (OGG1) were elevated by H2O2, and DMF treatment showed similar but pronounced effects through activation of Nrf2. To determine the relationship of Nrf2 and the generation of antioxidants, siRNAs were used and quantitative RT-PCR were conducted. Decreased expression of Nrf2 resulted in a decreased level of these antioxidant mRNA. Intracellular levels of ROS were investigated by fluorescence of 8-hydroxy-2'-deoxyguanosine and fluorescent dye, 2',7'-dichlorodihydrofluorescein diacetate after H2O2 and/or DMF treatment, and DMF treatment quenched intracellular ROS generation by H2O2. These results show that activation of Nrf2 might lead to alleviate OS in human GCs, and this could provide novel insight to conquer the age-related fertility decline that is mainly attributed to the accumulation of aberrant OS.

The effect of dehydroepiandrosterone supplementation on ovarian response is associated with androgen receptor in diminished ovarian reserve women

BackgroundDiminished ovarian reserve(DOR) is associated with female infertility and poor response to ovarian stimulation. Our objective was to assess the effect of dehydroepiandrosterone(DHEA) on DOR women and to explore whether the improvement of ovarian response after DHEA supplementation was dependent on the expression levels of androgen receptor(AR).MethodsA prospective cohort study was performed in the Department of Human Reproductive Medicine, Beijing Obstetrics and Gynecology Hospital during August 2014 to August 2016. 103 DOR women who completed the study were divided into the DHEA group (n = 53), which received DHEA supplementation (25 mg three times a day) for 8 weeks, and the control group (n = 50), which did not receive DHEA, before the IVF cycles. Serum hormone levels(FSH, LH, E2, T, DHEAs, AMH, INHB), antral follicle count(AFC) and the expression of AR and FSH receptor(FSHR) in granulosa cells(GCs) were measured, meanwhile ovarian response parameters and IVF outcomes were compared. The GCs from another 36 DOR women were cultured with different concentrations of DHEA in vitro. Then, we compared the expression of AR and FSHR in GCs according to the different numbers of oocytes retrieved both in DHEA and control group.ResultsIn the present study, DHEA supplementation resulted in significantly higher levels of serum T(P = 0.047), DHEAs(P = 0.019) and AR mRNA expression in GCs(P = 0.049). In vitro experiment, the protein and mRNA expression of AR and FSHR in the preovulatory GCs were significantly increased in response to DHEA supplementation(P <0.05). No significant differences were found in ovarian reserve, ovarian response, or IVF outcomes between the two groups. Subgroup analyses showed the levels of AR and FSHR mRNA in GCs were significantly increased in DHEA group with ≥5 oocytes retrieved(P <0.05).ConclusionDHEA supplementation can increase the expression of AR in preovulatory GCs both in vivo and in vitro. The selective beneficial effects of DHEA supplementation on ovarian response in DOR women may depend on the increasing expression of AR and FSHR in GCs.Trial registrationThe Chinese Clinical Trial Registry (ChiCTR-IPR-15006126). Retrospectively Registered 19 March 2015.

Self-organized amniogenesis by human pluripotent stem cells in a biomimetic implantation-like niche.

Amniogenesis-the development of amnion-is a critical developmental milestone for early human embryogenesis and successful pregnancy. However, human amniogenesis is poorly understood due to limited accessibility to peri-implantation embryos and a lack of invitro models. Here we report an efficient biomaterial system to generate human amnion-like tissue invitro through self-organized development of human pluripotent stem cells (hPSCs) in a bioengineered niche mimicking the invivo implantation environment. We show that biophysical niche factors act as a switch to toggle hPSC self-renewal versus amniogenesis under self-renewal-permissive biochemical conditions. We identify a unique molecular signature of hPSC-derived amnion-like cells and show that endogenously activated BMP-SMAD signalling is required for the amnion-like tissue development by hPSCs. This study unveils the self-organizing and mechanosensitive nature of human amniogenesis and establishes the first hPSC-based model for investigating peri-implantation human amnion development, thereby helping advance human embryology and reproductive medicine.

FEATURES OF FOLLICULOGENESIS, OOGENESIS AND EMBRYOGENESIS OF WOMEN IN ART CYCLES ON THE BACKGROUND PATHOLOGY OF THE THYROID GLAND

Introduction. In recent years, the relationship becomes actuality problem of violations of reproductive and thyroid glands (thyroid). On the one hand, this is due to the fact that the prevalence of infertility in marriage is 13-15% and in spite of the advancements in the field of human reproductive medicine, there is no downward trend. On the other hand, thyroid diseases occupy first place in the structure of endocrine pathology among women of reproductive age. Purpose - improving the efficiency restore reproductive function in women with infertility in thyroid pathology background. Results and discussion. The features of folliculogenesis, oogenesis and embryogenesis of women in art cycles on the background of thyroid disease. The detection rate of low level AMH in patients with pathology of the thyroid was 2.5 times above, than in women without thyroid pathology. In women young and middle age with disorders of the thyroid was dominated by low and medium levels of AMH, while the high level of AMH was only every third women aged 20-27 years (36,4%) and every 6 patients in the age 28-35 years, indicating a decrease in ovarian reserve in women with thyroid pathology. Conclusions. The studies indicate a reduction in the basic parameters ovarian reserve in patients with thyroid disorders compared with a group of women without thyroid pathology, namely, the significant reduction in average concentrations of AMH and FSH levels rise in average decreased ovarian volume (<5 cm 3) was diagnosed in 3 7 times more frequent in the group of patients with thyroid disorders. The normal ovarian volume (5 -10 cm 3) in the first group experienced 1.5 times less than in the second group. The average number of antral follicles in the ovaries of patients with thyroid disorders was significantly lower than in the control group. Thus, women with thyroid disorders in assisted reproduction cycles found significantly lower rates folliculogenesis and embryogenesis compared with women without thyroid disease.

From Ethical Exceptionalism to Ethical Exceptions: The Rule and exception Model and the Changing Meaning of Ethics In German Bioregulation.

Germany is an interesting case with respect to the governance of reprogenetics. It has a strong profile in the technosciences and high aims regarding the global bioeconomy, yet her regulation of human genetics, reproductive medicine and embryo research has for a long time been rather restrictive. German biopolitical exceptionalism has often been explained by reference to Catholicism and the legacy of the Nazi past. The Germans, so goes the common story, have learnt the lessons of history and translated them into unconditional respect for human dignity, which in turn translates into unconditional protection of human life, including the human embryo, and the firm repudiation of any eugenic distinction between 'life worth to live' and 'life not worth to live'. This, however, is not the whole story. Alongside deontological strictness we find another strand of governing body politics and reprogenetics in Germany, the rule-and-exception model, running from the mid-1970s abortion law via the 2002 Stem Cell Act to the 2011 regulation of pre-implantation genetic diagnosis. In contrast to the former, that strongly draws on Kant and his concept of human dignity, the latter bears resemblances to Carl Schmitt's concept of state of exception. The article will show that the rule-and-exception model builds the exception into the rule and transforms the meaning and mandate of ethics, namely from safeguarding ethical standards to deciding about the exception. Given that the exception has now tended to become the rule, the question is whether the lessons of history will govern German reprogenetics for much longer.

Cell death is involved in sexual dimorphism during preimplantation development

In bovine preimplantation development, female embryos progress at lower rates and originate smaller blastocysts than male counterparts. Although sex-specific gene expression patterns are reported, when and how sex dimorphism is established is not clear. Differences among female and male early development can be useful for human assisted reproductive medicine, when X-linked disorders risk is detected, and for genetic breeding programs, especially in dairy cattle, which requires female animals for milk production. The aim of this study was to characterize the development of female and male embryos, attempting to identify sex effects during preimplantation development and the role of cell death in this process. Using sex-sorted semen from three different bulls for fertilization, we compared kinetics of bovine sex-specific embryos in six time points, and cell death was assessed in viable embryos. For kinetics analysis, we detected an increased population of female embryos arrested at 48 and 120h.p.i., suggesting this time points as delicate stages of development for female embryos that should be considered for testing improvement strategies for assisted reproductive technologies. Assessing viable embryos quality, we found 144h.p.i. is the first time point when viable embryos are phenotypically distinct: cell number is decreased, and apoptosis and cell fragmentation are increased in female embryos at this stage. These new results lead us to propose that sex dimorphism in viable embryos is established during morula-blastocyst transition, and cell death is involved in this process.

Comparative intrauterine development and placental function of ART concepti: implications for human reproductive medicine and animal breeding.

The number of children conceived using assisted reproductive technologies (ART) has reached >5 million worldwide and continues to increase. Although the great majority of ART children are healthy, many reports suggest a forthcoming risk of metabolic complications, which is further supported by the Developmental Origins of Health and Disease hypothesis of suboptimal embryo/fetal conditions predisposing adult cardiometabolic pathologies. Accumulating evidence suggests that fetal and placental growth kinetics are important features predicting post-natal health, but the relationship between ART and intrauterine growth has not been systematically reviewed. Relevant studies describing fetoplacental intrauterine phenotypes of concepti generated by in vitro fertilization (IVF), intracytoplasmic sperm injection (ICSI) and somatic cell nuclear transfer (SCNT) in the mouse, bovine and human were comprehensively researched using PubMed and Google Scholar. Intrauterine growth plots were created from tabular formatted data available in selected reports. ART pregnancies display minor but noticeable alterations in fetal and placental growth curves across mammalian species. In all species, there is evidence of fetal growth restriction in the earlier stages of pregnancy, followed by significant increases in placental size and accelerated fetal growth toward the end of gestation. However, there is a species-specific effect of ART on birthweights, that additionally vary in a culture condition-, strain-, and/or stage at transfer-specific manner. We discuss the potential mechanisms that underlie these changes, and how they are affected by specific components of ART procedures. ART may promote measurable alterations to intrauterine growth trajectory and placental function. Key findings include evidence that birthweight is not a reliable marker of fetal stress, and that increases in embryo manipulation result in more deviant fetal growth curves. Because growth kinetics in early life are particularly relevant to adult metabolic physiology, we advise more rigorous assessment of fetal growth and placental function in human ART pregnancies, as well as continued follow-up of ART offspring throughout post-natal life. Finally, strategies to minimize embryo manipulations should be adopted whenever possible.

SIRT3 positively regulates the expression of folliculogenesis- and luteinization-related genes and progesterone secretion by manipulating oxidative stress in human luteinized granulosa cells.

SIRT3 is a member of the sirtuin family and has recently emerged as a vital molecule in controlling the generation of reactive oxygen species (ROS) in oocytes. Appropriate levels of ROS play pivotal roles in human reproductive medicine. The aim of the present study was to investigate SIRT3 expression and analyze the SIRT3-mediated oxidative response in human luteinized granulosa cells (GCs). Human ovarian tissues were subjected to immunohistochemical analysis to localize SIRT3 expression. Hydrogen peroxide and human chorionic gonadotropin were used to analyze the relationship between ROS and SIRT3 by quantitative RT-PCR and Western blotting. Intracellular levels of ROS were investigated by fluorescence after small interfering RNA-mediated knockdown of SIRT3 in human GCs. To uncover the role of SIRT3 in folliculogenesis and luteinization, mRNA levels of related genes and the progesterone concentration were analyzed by quantitative RT-PCR and immunoassays, respectively. We detected the expression of SIRT3 in the GCs of the human ovary. The mRNA levels of SIRT3, catalase, and superoxide dismutase 1 were up-regulated by hydrogen peroxide in both COV434 cells and human GCs and down-regulated by human chorionic gonadotropin. Knockdown of SIRT3 markedly elevated ROS generation in human GCs. In addition, SIRT3 depletion resulted in decreased mRNA expression of aromatase, 17β-hydroxysteroid dehydrogenase 1, steroidogenic acute regulatory protein, cholesterol side-chain cleavage enzyme, and 3β-hydroxysteroid dehydrogenase in GCs and thus resulted in decreased progesterone secretion. These results have the important clinical implication that SIRT3 might play a positive role in the folliculogenesis and luteinization processes in GCs, possibly by sensing and regulating the generation of ROS. Activation of SIRT3 function might help to sustain human reproduction by maintaining GCs as well as oocytes.

High Fertility: Two outbred mouse models exhibit different molecular and physiological strategies warranting improved fertility

Animal models are valuable tools in fertility research. Worldwide > 400 transgenic or knockout mouse models are available showing a reproductive phenotype; almost all of them exhibit an infertile or at least subfertile phenotype. In contrast, animal models revealing an improved fertility phenotype are barely described. Here we show data on two outbred mouse models showing a 'high fertility' phenotype. These mouse lines have been generated via selection over a time period of > 40 years and 161 generations. During this selection period the number of offspring per litter and the total birth weight of the entire litter nearly doubled. Concomitantly with the increased fertility phenotype several endocrine parameters (e.g. serum testosterone concentrations in male animals), physiological parameters (e.g. body weight, accelerated puberty, life expectancy) and behavioural parameters (e.g. behaviour in an open field, endurance fitness on a treadmill) have been altered. We elucidate that the two independently bred high fertility mouse lines warranted their improved fertility phenotype using different molecular and physiological strategies. The fertility lines display male- as well as female-specific characteristics. These genetically heterogeneous mouse models provide new insights into molecular and cellular mechanisms that enhance fertility. In view of decreasing fertility in men it will therefore be a precious information source for human reproductive medicine.

Abstracts

Reproduction in Domestic AnimalsVolume 49, Issue s1 p. 1-52 AbstractFree Access Abstracts First published: 06 February 2014 https://doi.org/10.1111/rda.12292Citations: 1AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Citing Literature Volume49, Issues1Special Issue: 47th Annual Conference of Physiology and Pathology of Reproduction, 39th Joint Conference on Veterinary and Human Reproductive Medicine, Giessen, 27-28 February 2014March 2014Pages 1-52 RelatedInformation

Prooxidant Effects of Verbascoside, a Bioactive Compound from Olive Oil Mill Wastewater, onIn VitroDevelopmental Potential of Ovine Prepubertal Oocytes and Bioenergetic/Oxidative Stress Parameters of Fresh and Vitrified Oocytes

Verbascoside (VB) is a bioactive polyphenol from olive oil mill wastewater with known antioxidant activity. Oxidative stress is an emerging problem in assisted reproductive technology (ART). Juvenile ART is a promising topic because, in farm animals, it reduces the generation gap and, in human reproductive medicine, it helps to overcome premature ovarian failure. The aim of this study was to test the effects of VB on the developmental competence of ovine prepubertal oocytes and the bioenergetic/oxidative stress status of fresh and vitrified oocytes. In fresh oocytes, VB exerted prooxidant short-term effects, that is, catalase activity increase and uncoupled increases of mitochondria and reactive oxygen species (ROS) fluorescence signals, and long-term effects, that is, reduced blastocyst formation rate. In vitrified oocytes, VB increased ROS levels. Prooxidant VB effects in ovine prepubertal oocytes could be related to higher VB accumulation, which was found as almost one thousand times higher than that reported in other cell systems in previous studies. Also, long exposure times of oocytes to VB, throughout the duration of in vitro maturation culture, may have contributed to significant increase of oocyte oxidation. Further studies are needed to identify lower concentrations and/or shorter exposure times to figure out VB antioxidant effects in juvenile ARTs.

Endurance Exercise and Conjugated Linoleic Acid (CLA) Supplementation Up-Regulate CYP17A1 and Stimulate Testosterone Biosynthesis

A new role for fat supplements, in particular conjugated linoleic acid (CLA), has been delineated in steroidogenesis, although the underlying molecular mechanisms have not yet been elucidated. The aims of the present study were to identify the pathway stimulated by CLA supplementation using a cell culture model and to determine whether this same pathway is also stimulated in vivo by CLA supplementation associated with exercise. In vitro, Leydig tumour rat cells (R2C) supplemented with different concentrations of CLA exhibited increasing testosterone biosynthesis accompanied by increasing levels of CYP17A1 mRNA and protein. In vivo, trained mice showed an increase in free plasma testosterone and an up-regulation of CYP17A1 mRNA and protein. The effect of training on CYP17A1 expression and testosterone biosynthesis was significantly higher in the trained mice supplemented with CLA compared to the placebo. The results of the present study demonstrated that CLA stimulates testosterone biosynthesis via CYP17A1, and endurance training led to the synthesis of testosterone in vivo by inducing the overexpression of CYP17A1 mRNA and protein in the Leydig cells of the testis. This effect was enhanced by CLA supplementation. Therefore, CLA-associated physical activity may be used for its steroidogenic property in different fields, such as alimentary industry, human reproductive medicine, sport science, and anti-muscle wasting.

MHR welcomes high-quality basic reproductive research around pregnancy

The eight publications in the November issue of the journal (one review and seven original articles) are all about pregnancy. To some of our readers this will be a surprise—there is a popular myth that Molecular Human Reproduction (MHR) loses interest in reproduction after implantation. The papers we publish this month demonstrate that this just is not true. We consider scientific reports on topics straddling all of pregnancy. Thus, our scope is wider than our sister journal, Human Reproduction. Indeed, a review of Molecular Human Reproduction publications over the last 5 years demonstrate that 33% are identified by the search term ‘pregnancy’—a situation that we very much hope will continue. So what are the criteria for publication in Molecular Human Reproduction? Simply put, it is excellence in basic reproductive science relevant to human reproductive medicine. We favour work describing robust mechanistic studies, preferably done using primary human tissues (either exclusively, or to confirm key findings made using cell lines). We are also interested in work that reports novel findings, utilizes novel techniques and—if relevant—describes in vivo studies. So how do the collection of articles published this month qualify, and how do they advance the field of basic reproductive research around pregnancy?

Certified Angus, Certified Patriot: Breeding, Bodies, and Pedigree Practices

The pedigree chart is a cornerstone technology for producing bodies and value in livestock pure breeding. It organizes a cluster of processes, technologies, and discourses gathered under the rubric ‘pedigree practices’. Angus breeders commonly use artificial insemination to import performance ‘genetics’ into their herds, using the ‘expected progeny differences’ predicted by massive pedigree databases that now also contain phenotype data reported by cattle growers. Discourses of biological inheritance, good breeding, and pedigrees arose in the eighteenth century, concomitant with a fascination with races, species, and other biological kinds. A case study from Angus cattle breeding illustrates pedigree practices and the bodies made through them, showing how information and computing technologies, assisted reproductive technologies, and discourses of good breeding, purity, health, and disease leveraged a single bull and the two genetic diseases he carried into the pedigrees of up to 10% of the Angus herd. Technologies now widely used in human reproductive medicine were developed for use in livestock animals, especially cattle, extending a long relationship between cows and humans. While the development of these pedigree practices represents increased control over animal reproduction and bodies, it has also been instrumental in rendering all animal bodies, including human bodies, not only more technically but also more rhetorically available for reproductive interventions. These interventions may amplify anxieties about health, species, breed, and kin while also providing opportunities for contesting the boundaries of these nature–cultural categories.

Il caso Italia: medicina riproduttiva e obiezione di coscienza

The subject of this article is the Italian law in European context, regulating abortion and the conscientious objection by medical staff. The methodology is one developed by Jasanoff in Designs of Nature. After describing the historical, cultural and national policy, the article proceeds with the analysis of art. 9 of Law May 22, 1978 194, which regulates conscientious objection on the part of medical personnel to the voluntary interruption of pregnancy. The issue of conscientious objection is framed also in the European context. In this regard, the analysis focuses on the document that the Social Health and Family Issues Committee presented to the European Parliamentary Assembly, entitled Women's access to lawful medical care: the problem of unregulated use of conscientious objection. In this document , Italy is seen as an emblematic case of the problematic legal institution of conscientious objection, as confirmed by Table 28 of the Report 2012 of the Ministry of Health on the implementation of Law 194, according to which the percentage of objection exceeds 69%. There is a big focus on the comparison between the levels of involvement and action of different political, institutional, social and cultural entities: the role of the Italian State in producing standards and knowledge in terms of human reproductive medicine, as well as the role of ethics committees and epistemic national communities. The analysis focuses on the development of European and international law, on the contributions of the categories of professionals, on the opinions of bioethicists about the issue of conscientious objection to medical techniques relevant for and moral philosophy, such as the interruption of pregnancy. For this purpose, the article analysed the different perspectives expressed by professional associations, such as the League of not-objector gynecologists called LAIGA, and the committee of bioethicists, such as the National Bioethics Committee, which, in the document of 30 July 2012, entitled Conscientious Objection and bioethics declared to be favour of it.

Il caso Italia: medicina riproduttiva e obiezione di coscienza

The subject of this article is the Italian law in European context, regulating abortion and the conscientious objection by medical staff. The methodology is one developed by Jasanoff in Designs of Nature. After describing the historical, cultural and national policy, the article proceeds with the analysis of art. 9 of Law May 22, 1978 194, which regulates conscientious objection on the part of medical personnel to the voluntary interruption of pregnancy. The issue of conscientious objection is framed also in the European context. In this regard, the analysis focuses on the document that the Social Health and Family Issues Committee presented to the European Parliamentary Assembly, entitled Women's access to lawful medical care: the problem of unregulated use of conscientious objection. In this document , Italy is seen as an emblematic case of the problematic legal institution of conscientious objection, as confirmed by Table 28 of the Report 2012 of the Ministry of Health on the implementation of Law 194, according to which the percentage of objection exceeds 69%. There is a big focus on the comparison between the levels of involvement and action of different political, institutional, social and cultural entities: the role of the Italian State in producing standards and knowledge in terms of human reproductive medicine, as well as the role of ethics committees and epistemic national communities. The analysis focuses on the development of European and international law, on the contributions of the categories of professionals, on the opinions of bioethicists about the issue of conscientious objection to medical techniques relevant for and moral philosophy, such as the interruption of pregnancy. For this purpose, the article analysed the different perspectives expressed by professional associations, such as the League of not-objector gynecologists called LAIGA, and the committee of bioethicists, such as the National Bioethics Committee, which, in the document of 30 July 2012, entitled Conscientious Objection and bioethics declared to be favour of it.

Initial characterization of an outbreed mouse model for male factor (in)fertility

We analysed an outbreed mouse line which was selected for the phenotype 'high fertility' for 158 generations. During this selection period the mouse strain increased the number of offspring per litter from 10.4 to 17.1 and the total litter weight up to ~160%. In this study, we initially characterize the reproductive phenotype of high fertility males. Surprisingly, male bucks of the fertility line (FL1) show reduced percentage of motile and progressive motile spermatozoa; however, other sperm motility characteristics (e.g. velocity parameters) are improved compared with an unselected control line. Cytometrical investigation of the testicular cell-type composition indicated a significant increased concentration of diploid cells by a concomitant reduction in haploid cells in the testicular parenchyma of FL1. Furthermore, total testosterone concentrations in blood are dramatically increased in FL1 (>20ng/mL). In line with increased testosterone levels, we observed increased expression rates of steroidogenic key enzymes Cyp11 and Cyp17 from FL1 testis samples. These data indicate that FL1 males have a manifest 'high fertility phenotype'. Diallelic crosses imply that male-only contribution largely determines the reproductive outcome in cross-breeding experiments. FL1 therefore is a promising model for future investigations on male factor (in)fertility. Our observation might also offer valuable cues for human reproductive medicine.

Acute alcohol exposure, acidemia or glutamine administration impacts amino acid homeostasis in ovine maternal and fetal plasma

Fetal alcohol syndrome (FAS) is a significant problem in human reproductive medicine. Maternal alcohol administration alters maternal amino acid homeostasis and results in acidemia in both mother and fetus, causing fetal growth restriction. We hypothesized that administration of glutamine, which increases renal ammoniagenesis to regulate acid-base balance, may provide an intervention strategy. This hypothesis was tested using sheep as an animal model. On day 115 of gestation, ewes were anesthetized and aseptic surgery was performed to insert catheters into the fetal abdominal aorta as well as the maternal abdominal aorta and vena cava. On day 128 of gestation, ewes received intravenous administration of saline, alcohol [1.75 g/kg body weight (BW)/h], a bolus of 30 mg glutamine/kg BW, alcohol + a bolus of 30 mg glutamine/kg BW, a bolus of 100 mg glutamine/kg BW, alcohol + a bolus of 100 mg glutamine/kg BW, or received CO2 administration to induce acidemia independent of alcohol. Blood samples were obtained simultaneously from the mother and the fetus at times 0 and 60 min (the time of peak blood alcohol concentration) of the study. Administration of alcohol to pregnant ewes led to a reduction in concentrations of glutamine and related amino acids in plasma by 21-30%. An acute administration of glutamine to ewes, concurrent with alcohol administration, improved the profile of most amino acids (including citrulline and arginine) in maternal and fetal plasma. We suggest that glutamine may have a protective effect against alcohol-induced metabolic disorders and FAS in the ovine model.