In our environment there are many compounds which can negatively influence humans and wildlife. Every day, a vast number of environmental pollutants are released into our environment and there is no way to avoid their exposure. Some of these compounds can even mimic endogenous hormones and interfere with our endocrine system (so called endocrine disruptors), which is the key regulatory system controlling almost all physiological processes in human and animal bodies. Also the reproductive system is largely regulated by various hormones, and their proper function is crucial for gamete formation, fertilization and embryo development. Environmental pollutants are therefore considered as one of the possible causes of increased infertility in human population. This prompted us to study the effect of two endocrine disruptors (tetrabromobisphenol A - TBBPA, and zearalenone - ZEA) on the male mouse reproductive system in vivo. According to our results, TBBPA is able to induce apoptosis as well as changes in the expression of selected testicular genes and sperm protamination. Our results also suggest that permanent exposure to TBBPA slightly enhances its effect in the next generation, depending on whether the parents have been affected or not. We hypothesized that differential protamination of the sperm DNA...

Assisted reproductive technologies are the phenomenon of the nowadays. With respect to the growing possibilities of the medical science and its impact on lives of individuals it is very important to consider the legal regulation of assisted reproduction. The aim of the thesis is to analyse the law of assisted reproduction in the Czech Republic in wider context in three chapters. The first chapter provides the reasons why it is necessary to regulate assisted reproduction, even though the law does not and should not interfere with the natural reproduction, reflecting the power of the modern reproductive technologies and new concerns it raises. Moreover, it focuses on the issues which need to be remembered when setting the legal rules in general, including the ethical aspects and access to treatment, considering the concept of reproductive autonomy of individuals. The law of assisted reproduction in the Czech Republic is described in the second chapter, which includes the throughout analysis of the Czech law. The thesis includes the explanation of essential legal definitions as assisted reproduction and artificial insemination and summarises the legal conditions, under which the treatment by assisted reproduction technologies can be accessed including the financial issues. Moreover, surrogacy and its...

M.Cur. / With this research it was endeavoured to gain more knowledge about the experiences of women during the treatment of infertility. The midwife and other medical staff involved with the treatment of infertility can use this knowledge to have better insight into the total problem of these women and also care for and support them better. A study of the literature was done to determine what the experts have already ascertained of this phenomenon. An in-depth study of the phenomenon was undertaken in which the autobiographical method was mostly used. During April 1989 unstructured interviews were conducted with six selected infertile women living in the witwatersrand area. From the findings of this research it appears that infertile women have unique experiences of their infertility and the treatment of it. Universal experiences, ego pain and disappointment were identified, although the causes of these experiences sometimes differed. Recommendations were made on how the midwife can contribute by filling the gap (established during the research) in the care in this field. Suggestions were also made with regard to further research on this phenomenon.

Philosophiae Doctor - PhD / The metabolic syndrome (MetS) is a collection of various metabolic, hormonal and immunological risk factors that cluster together, closely related to poorly understood phenomena such a hyperinsulinaemia (insulin resistance), hyperleptinaemia (leptin resistance), a low grade, systemic and chronic inflammation and, in males, hypogonadism. Infertility is increasing globally, and male factor infertility accounts for a large percentage of couples who are not able to conceive. The relationship between components of MetS and male reproductive health is not clear, and requires further investigation, as does the impact of MetS on male reproductive health in a case controlled study. The impact of hyperinsulinaemia, hyperleptinaemia and inflammatory cytokines on the male reproductive tract also requires investigation. Furthermore, it is hypothesised that these phenomena negatively impact steroidogenesis cascades. In order to investigate this, a case controlled study and TM3 Leydig cell culture experiments were designed.Participants were recruited from public advertisements, and screened for strict exclusion criteria, including acute or chronic inflammation, hormonal treatments, vasectomy and leukocytospermia (> 106/ml). Following clinical diagnostics, 78 males were either placed into a control group (CG) or the MetS group, with numerous parameters compared between them. Serum was assayed for routine risk markers including HDL cholesterol, triglycerides, glucose and C-reactive protein (CRP). Saliva was assayed for free testosterone and progesterone. Semen samples underwent semen analysis for ejaculation volume, sperm concentration and motility, vitality, morphology and leukocyte concentration, in addition to mitochondrial membrane potential (MMP) and DNA fragmentation (DF). Both serum and seminal fluid were further assayed for insulin, leptin, tumour necrosis factor-alpha (TNF ) and interleukins 1-beta (IL1 ), 6 (IL6) and 8 (IL8). Glucose was also assayed in seminal fluid. Separately, hCG stimulated TM3 Leydig cells were exposed to varying concentrations of insulin (0.01, 0.1, 1 & 10 pg/ml), TNF , IL1 , IL6 and IL8 (0.1, 1, 10 & 100 pg/ml) for 48 hours at optimal cell culture conditions. TM3 cell viability, protein concentration and testosterone and progesterone concentrations were assessed.XXII Results indicated that males in the MetS group (n=34) had significantly increased body mass index, waist circumference, blood pressure, triglycerides, glucose, and Creactive protein (CRP) with decreased HDL cholesterol, as compared to the CG. Furthermore, ejaculation volume, sperm concentration, total sperm count, progressive and total motility were significantly decreased in the MetS group, and sperm with abnormal MMP and DF were increased in this group. No difference was found for morphology. Serum and seminal insulin, leptin, TNF , IL1 , IL6 and IL8 were all significantly increased in the MetS group. Both testosterone and progesterone were also significantly decreased in the MetS group. Insulin increased testosterone and decreased progesterone in the TM3 cells. TNF , IL1 and IL6 all decreased testosterone and progesterone concentrations and TM3 cell viability. IL8 increased TM3 cell viability and decreased progesterone, will no effect on testosterone. These results suggest MetS is associated with decreased fertility potential in males. Furthermore, a significant increase in seminal insulin, leptin, TNF , IL1 , IL6 and IL8 suggests local reproductive tract inflammation in the absence of leukocytospermia. Strong correlations between serum and seminal insulin, leptin, TNF , IL1 , IL6 and IL8, as well as serum CRP, imply that these systemic phenomenons are related to the reproductive tract changes observed. Therefore, the underlying pathophysiology of MetS negatively affects male reproduction, in addition to general health and wellbeing. A decrease in progesterone and testosterone suggests a collapse in steroidogenesis cascades. Additionally, inflammation, increased leptin and insulin resistance likely contribute to this collapse in steroidogenesis based on TM3 cell culture experiments. These results provide novel avenues for further investigations.

Magister Scientiae (Medical Bioscience) - MSc(MBS) / There is increasing concern worldwide by researchers with regards to the assessing of safety and therapeutic consumption of the plants used in traditional medicine. Date palm pollen (DPP) and flax seed have been used traditionally to improve fertility in Libya. DPP extracts have shown several reproductive beneficial effects. In vivo, studies have revealed the ability of DPP to increase sperm concentrations, ameliorate the testicular toxicity induced by cadmium and lead, raise testosterone, as well as LH and FSH hormone levels. Flax seed phytochemical analysis showed lots of valuable components such as lignans and α linolenic acid to which were attributed its positive health effects like antitumor, antioxidant and protective effects against coronary heart diseases. Moreover, flax lignans have both estrogenic and antiestrogenic potential. This study was aimed at testing the effects of Libyan DPP and flax seed on the Sertoli (TM4) cell line and human breast adenocarcinoma (MCF - 7) cell line. Different concentrations (0.01, 0.1, 1, 10, 100 and 1000 μg/ml) of ethanolic extracts of DPP and flax seed, respectively, were used to assess the morphology of TM4 and MCF - 7 cells after 24 and 72 hours exposure. Mitochondrial dehydrogenase activity as a marker of cell viability was measured by MTT assay after 24 and 72 hours exposure. Apoptotic effects were assessed by flow cytometeric APO percentage assay. TM4 cell production of Inhibin - B hormone and GGT enzyme activity under the effects of DPP or flax seed was determined by use of ELISA kits. Transepithelial electrical resistance (TEER) assay were used to detect the effect of DPP or flax seed on TM4 cell monolayer integrity. Finally the plants potential phytoestrogenic activity was determined by use of E - SCREEN assay in MCF – 7 breast cancer cells. Higher concentrations of DPP significantly increased the activity of mitochondrial dehydrogenase enzyme of TM4 cells after 24 hours associated with increasing cell number as detected in a microphotograph. Flax seed concentrations less than 100 μg/ml reduced TM4 cell viability but there were no morphological changes visible after 24 hours. MCF - 7 cells viability was reduced after 24 and 72 hours treatment with DPP and flax seed. DPP concentrations beyond 1 μg/ml significantly raised the TEER of TM4 monolayer over 72 hours while flax seed treatments caused a significant increase only after 72 hours of exposure. TM4 cells GGT activity increased significantly after exposure to higher concentrations of DPP and all flax seed concentrations. Significant stimulatory effects of all the concentrations of DPP or flax seed on TM4 inhibin - B hormone productions have been detected. Apoptotic studies showed no significant changes. E - SCREEN assay resulted in significant reduction in MCF - 7 proliferation rate under the effect of low concentrations of DPP or flax seed. Higher concentrations of the plant extracts, however, stated to increase MCF – 7 cell proliferation, this exerts weak estrogenic activities. In conclusion, the main finding of this study is that DPP and flax seed showed stimulatory effects on TM4 cells proliferation. The resistivity of TM4 cells monolayer which reflect the integrity of blood – testis barrier (BTB) was also significantly increased as well as inhibin - B production and GGT enzyme activity. In addition DPP and flax seed respectively showed inhibitory effects on MCF - 7 cells viability. This study indicated that DPP or flax seed may enhance spermatogenesis through their stimulatory action on Sertoli cells. Moreover, both plants could reduce breast cancer cells viability. However, further investigations are required to elucidate the exact mechanisms behind these obtained findings.

Fertility lies at the heart of Life: it re-invigorates and regenerates. It is one of the most intimate areas of human existence. Worldwide, infertility is on the increase. However, advances in biomedical technologies, such as in vitro fertilisation (IVF) give hope to those who are suffering from infertility. At the same time it brings the question of moral responsibility into focus. The availability of donor sperm and eggs, coupled with greater recognition of the rights of lesbians and gays to become parents either by adoption, utilising donor material, or openly exercising their rights as already existing parents, have complicated previous more conservative understandings of what ‘family’ means. The epistemological point of departure of this research is described in Chapter 3 and is positioned within postfoundational practical theology, with an emphasis on critical emancipative feminism. The underlying research methodology is the narrative approach, embedded in social constructionism. The research explores the narratives of four couples. These co-researchers, affected by infertility and childlessness, share their stories of surrogacy, secondary infertility, gay parenting and miscarriage that are presented in Chapters 4 to 7. In Chapter 2 the researcher includes her story as someone living with infertility. Particular attention was paid to understand and develop insights concerning preferred lives of fruitfulness in spite of infertility and childlessness. The explored narratives revealed varying discourses that are introduced through out the thesis, but are specifically integrated in Chapter 8. In the final chapter the author reflects critically on the research and writing process as a whole. / Thesis (PhD)--University of Pretoria, 2009. / Practical Theology / unrestricted

This study explores the experiences of infertile married African women in South Africa. The study explores the women’s emotional experiences, their experiences of their relationships and the possible ways in which their culture may have influenced their experiences. The literature review gives an overview of female infertility, motherhood and feminism- from its conception to its current status in South Africa. The study is carried out in a clinical setting where the participants are already attending an infertility clinic for treatment. Six black women participate in this study and they are all married and experiencing primary infertility. This study does not include women experiencing secondary infertility because although they may presently be infertile, they have previously experienced one or more successful pregnancies. Therefore, primary infertility is chosen as the focus of this study because unlike secondary infertile women, primary infertile women have never experienced pregnancy and childbirth and thus their infertility is more noticeable and severe. The study is qualitative in nature and it is conducted within a feminist social constructionist research paradigm. This research paradigm is deemed to be more relevant in researching an issue pertaining to women, because feminist research enables social scientists to explore women’s social history, women’s perception of their own situation, their own subordination and their own resistance (Mies, 1993). Through social constructionism, the women are each able to make meaning out of their experiences and to include the influences of their culture, gender and social context in the construction of their experiences. Furthermore, since feminist research perspectives have supported the process of telling sensitive human stories as research, a narrative approach is employed in gathering information regarding the women’s experiences of being infertile. Thus, through telling their own self-narratives, the women are able to relate their experiences as well as the events that took place in their lives regarding their infertility. Structural narrative analysis as well as content narrative analysis are employed in the analysis of the women’s narratives. What emerges from their stories is the hope and the subsequent disappointment that follows from their countless visits and consultations with different health professionals, both western and traditional, as they search for pregnancy. It also emerges that in addition to their personal experiences, these women’s marital and familial relationships, as well as their extended social relationships, also contribute to the emotional distress that they experience as a result of their infertility. Although most of the women mainly report negative experiences, there are also some who report positive experiences and some form of support in their lives. It is hoped that the results obtained from this study will enable psychologists to intervene effectively and to work collaboratively with other health professionals towards delivering services aimed at assisting infertile African women medically, socially and psychologically. / Dissertation (MA (Clinical Psychology))--University of Pretoria, 2008. / Psychology / unrestricted

La spermatogenèse est un processus complexe qui dépend de la coopération de nombreux gènes. Son produit final le spermatozoïde, est un sujet d’étude idéal car il renferme à la fois des indices d’événements passés ainsi que des informations qui seront transmises à l'ovocyte lors de la fécondation. L'identification de nouveaux acteurs de la spermatogenèse, des modifications spécifiques de l'ADN du sperme ou la présence de transcrits spécifiques pourraient servir comme biomarqueurs dans le diagnostic de l’infertilité. Cette thèse avait pour but d’analyser le génome, le transcriptome et l’épigénome de spermatozoïdes dans le contexte de l'infertilité masculine. Nous avons identifié de nouvelles causes génétiques et confirmé la présence d'anomalies de méthylation dans le sperme d'hommes infertiles. Nous avons découvert 20 mutations dans le gène SOX8, chez des patients atteints de trouble du développement sexuel ou d'infertilité masculine ou féminine, qui apparaît comme un régulateur du développement et de la fonction gonadique. Par séquençage d’exome, une mutation dans le gène ATAD2 modeleur de la chromatine spécifique de la lignée germinale mâle fut également identifiée. Par RNA-seq et MeDIP-chIP du sperme d’hommes fertiles et infertiles, nous avons caractérisé la signature transcriptionnelle du sperme. La majorité des ARNs spermatiques humain est remarquablement conservée chez les mammifères placentaires suggérant des fonctions ancestrales importantes. Enfin, nos données transcriptomiques et épigénétiques tendent à indiquer qu’une expression et une régulation adéquates des gènes impliqués dans le remodelage de la chromatine constituent un facteur clé pour la fertilité masculine. / Spermatogenesis is a complex process which depends on the cooperation of many genes. The end-product, the spermatozoon, is an ideal subject for study since it carries both clues of the past events and information which will be transmitted to the oocyte at fertilization. The identification of main actors of spermatogenesis, specific modifications of sperm DNAs or sperm specific isoforms could improve our understanding of a such complex mechanism and could serve as a determination of biomarkers or diagnostic tools for fertility. The aim of the project was to go further three omes: genome, epigenome and transcriptome of mature human sperm in the context of male infertility. We identified new genetic causes of male infertility and confirmed the presence of methylation abnormalities in sperm cells of infertile men. Firstly, SOX8 gene was found mutated in a cohort of 20 patients with disorder of sex development and male or female infertility. Similarly, to NR5A1, SOX8 appears to be a novel regulator of gonadal development and function. Then by exome-sequencing, we identified a homozygous nonsense mutation in the male germline-specific chromatin modeler ATAD2. Furthermore, RNA-seq and MeDIP-chIP of sperm from fertile and infertile men along with bioinformatics analyzes of the generated data, enabled us to characterize more deeply the normal sperm transcriptional signature. We also found that the majority of human sperm RNAs are remarkably preserved in placental mammals suggesting crucial ancestral functions. Finally, proper expression and regulation of chromatin remodelers seem to be critical for male fertility, as revealed by both the transcriptomic and the epigenetic data.

Les translocations chromosomiques et les autres types de réarrangements peuvent, bien qu'associées à un phénotype normal, mener à la transmission d'un contenu génétique déséquilibré à la descendance. Il n'est pas possible de distinguer les chromosomes équilibrés des déséquilibrés, ce qui empêche toute sélection dans le cadre d'une fécondation in vitro (FIV). Nous avons ainsi mené une série de projets de recherche dont le but a été de mettre en évidence des caractéristiques spécifiques de ces spermatozoïdes déséquilibrés, afin de pouvoir les distinguer au cours d'une FIV. Nous avons montré que les spermatozoïdes déséquilibrés présentaient des taux de fragmentation de l'ADN plus élevés, étaient moins denses, et avaient un volume nucléaire supérieur. Ces constatations ont mené au développement d'une procédure de sélection des spermatozoïdes équilibrés chez les porteurs de réarrangements chromosomiques. En utilisant le hypo-osmotic swelling test (HOST), nous avons montré qu'une morphologie flagellaire spécifique était associée à un contenu chromosomique équilibré. Nous proposons d'utiliser cette procédure de sélection dans le cadre d'une ICSI, afin d'améliorer le pronostic reproductif chez les couples concernés. Nous proposons également d'évaluer la proportion de spermatozoïdes déséquilibrés chez chaque patient porteur de réarrangement chromosomique. En effet, bien que ce taux varie d'un patient à l'autre, il est stable dans le temps pour un patient donné. Il est de plus un élément déterminant dans le choix d'une des options de prise en charge : reproduction naturelle, insémination artificielle avec test de migration survie (TMS), ICSI avec sélection par HOST, et diagnostic préimplantatoire (DPI). / Chromosomal translocations and other balanced rearrangements, although usually associated with a normal phenotype, can lead to the transmission of an abnormal unbalanced genetic content to the offspring. Balanced and unbalanced spermatozoa are indistinguishable, making it impossible to select them prior to in vitro fertilization. We conducted a series of research projects aimed at evidencing specific characterics for unbalanced spermatozoa, in a way to ultimately distinguish them from balanced ones during in vitro fertilization. We showed that unbalanced spermatozoa had higher DNA fragmentation rates, were less dense, and that their nuclear volume was higher. This led to developing a selection procedure for balanced spermatozoa in rearrangement carriers. Using the hypo-osmotic swelling test (HOST), we showed that a specific flagellar morphology was associated with balanced chromosomal status. We suggest using this procedure prior to ICSI in order to improve the reproductive prognosis in those patients. We also suggest evaluating the proportion of unbalanced spermatozoa in every patient with a chromosomal rearrangement. Although this proportion varies among patients, it is stable over time for a given patient. We believe this is a decisive element in choosing between the different available options : natural conception, artificial insemination with discontinuous gradient centrifugation, ICSI with HOST-based selection, and pre-implantation genetic diagnosis (PGD).

The subject of my thesis is to analyze the evolution of the reproductive behavior of the Czech population to assess changes over time, especially after 1989 and to analyze the success of assisted reproduction in the Czech Republic in the period 2007 to 2013. It is divided into several parts. The first theoretical part describes the change in reproductive behavior after 1989, risk factors and causes of female and male infertility, possible tratments of diagnosed infertility and various methods of assisted reproduction. Another analytical part is dedicated to the evolution of fertility and the phenomenon of childlessness of young women. Some indicators are compared with selected European countries for an overall assessment. Subsequently are analyzed in detail the most widely used methods of assisted reproduction IVF, ICSI and kryoembryotransfer and of using indicators Pregnancy rate and Take-home baby rate is assessed their effectiveness. The thesis is supplemented by own research reproductive and family behavior on a selected sample of the Czech population.