Cell adhesion proteins osteopontin, CD44 and integrin alphaVbeta3 interact to form an adhesion complex between the embryo and endometrial surface forming an attachment that can lead to implantation. Whilst receptivity has been investigated extensively, the expression of this adhesion complex has yet to be studied simultaneously in the endometrium. This thesis establishes the expression of the adhesion complex in fertile and infertile endometrium. In addition the regulation of the adhesion complex components by distinct signalling pathways and the key regulators estrogen receptor, nuclear factor kappa B and signal transducer and activator of transcription 1 have been investigated in endometrial cell lines. Objectives: To establish the expression profile of adhesion complex components in samples obtained from fertile and infertile women. To model in vitro hormonal regulation of adhesion complex components to mimic estrogen and progesterone stimulus in the menstrual cycle. To determine if adverse environments common to poly cystic ovarian syndrome and endometriosis affect uterine expression of the adhesion complex via high glucose and pro-inflammatory cytokines. To investigate the direct regulation of Osteopontin and CD44 by estrogen and cytokine signalling through estrogen receptor ?, nuclear factor kappa B and signal transducer and activator of transcription 1. Methodology: Investigation of human biopsies and cell line models by immunohistochemistry, quantitative polymerase chain reaction, chromatin immunoprecipitation and enzyme linked immunosorbent assay. Conclusions: Adverse uterine environments including high glucose and pro-inflammatory cytokines may regulate the expression of the adhesion complex, and contribute to a lack of endometrial receptivity in endometriosis and poly cystic ovarian patients. CD44, ITGAV and ITGB3 levels may be used as markers for loss of receptivity in unexplained infertility.

Orientador: Adriana Orcesi Pedro / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-12T09:22:16Z (GMT). No. of bitstreams: 1 Silveira-Barbetti_CarolinaFernanda_D.pdf: 1494162 bytes, checksum: 743c1f97b1d23cbeab92921d0c8d7688 (MD5) Previous issue date: 2008 / Resumo: Introdução: A infertilidade masculina pode acometer entre 30% e 40% dos casais inférteis. Dentre esses fatores está a varicocele, que afeta 15% da população em geral e que nesses homens inférteis chega a 30%-40%, podendo estar relacionada com alterações na amostra seminal e conseqüente fertilidade. Objetivo: Comparar amostras seminais de homens férteis sem varicocele com homens inférteis com varicocele quanto a(os) parâmetros macroscópicos e microscópicos; recuperação espermática após swim-up; tipos de marcação dos receptores de D-manose em espermatozóides com acrossoma íntegro e ocorrência da reação acrossomal independente da ligação de D-manose em dois tempos. Sujeitos e Métodos: foi realizado um estudo corte transversal comparando o grupo de pacientes com queixa de infertilidade e varicocele (N=30) com o de homens férteis e sem varicocele (N=30) através do espermograma, processamento espermático e testes funcionais nas amostras recém-ejaculadas (zero hora) e após processamento (uma hora). Os dados foram analisados pelos testes exato de Fisher, Mann-Whitney, quiquadrado e análises repetidas de Friedman. Resultados: O grupo com varicocele apresentou alterações significativas em relação ao grupo-controle na análise seminal quanto à cor (p<0,01), viscosidade aumentada (p<0,01), menor concentração espermática (p<0,01), menor motilidade (A+B) (p<0,01), menor vitalidade (p<0,01), menor inchaço hiposmótico (p<0,01) e maior alteração na morfologia estrita de Kruger (p<0,01). Após capacitação espermática pela técnica de swim-up, o grupo com varicocele apresentou menor concentração espermática (p<0,01) e menor porcentagem de espermatozóides com expressões dos receptores de Dmanose e não reagidos acrossomicamente, independente do tempo (p<0,01). Não houve diferença significativa das amostras em cada grupo quando avaliadas separadamente, entre os dois momentos independentes do status acrossomal e da presença da expressão dos receptores para D-manose. Os quatro padrões de marcação com D-manose e acrossoma íntegro mais observados no grupo com varicocele foram cabeça manchada (21,1%), peça intermediária preenchida (13,1%), região equatorial preenchida (10,6%) e cabeça preenchida (9,8%). Conclusão: espermatozóides de homens com varicocele apresentaram alterações qualitativas, quantitativas e funcionais, podendo comprometer a fertilização espontânea. / Abstract: Introduction: Male infertility may affect 30% to 40% of infertile couples. Among this factors, varicocele affects 15% of the general population and may reach 30%-40% of infertile men. The varicocele to be able to exhibit disturbance in the semen specimens and fertility result.Objectives: To compare, at two time points, semen samples from fertile men and infertile men with varicocele with respect to macroscopic and microscopic parameters, sperm recovery after swim-up, patterns of labeling of D-mannose-ligand receptor in spermatozoids with intact acrosome and the occurrence of acrosomal reaction irrespective of the expression of D-mannose-binding sites. Subjects and methods: A crosssectional study was carried out to compare a group of patients with complaints of infertility and varicocele (n=30) and a control group of fertile men without varicocele (n=30) by performing semen analysis, sperm processing and functional tests carried out in recently ejaculated samples (time zero) and following processing (one hour). Statical analysis was performed through Fisher's exact test, chisquare test, Mann-Whitney test and Friedman repeated analysis of variance. Results: Significant alterations were found in the group of men with varicocele compared to the control group with respect to color (p<0.01), increased viscosity (p<0.01), lower sperm concentration (p<0.01), lower motility (A+B) (p<0.01), lower vitality (p<0.01), lower rates of hypoosmotic swelling (p<0.01), greater alterations in morphology according to Kruger's strict criteria (p<0.01). After performing swinup lower sperm concentration (p<0.01) and lower percentage of sperm cells expressing D-mannose binding sites, together with intact acrosomal status irrespective of time (p<0.01) in infertile men with varicocele. There was no statistically significant difference between the samples of each group when evaluated separately at the two time points, irrespective of acrosomal status and of D-mannose-ligand receptor expression. The four patterns of labeling of Dmannose binding sites and intact acrosome most observed in the varicocele group were: spotty pattern on the head (21.1%), uniform pattern on the mid-piece (13.1%), uniform pattern on the equatorial region (10.6%) and uniform pattern on the head (9.8%). Conclusions: Sperm from men with varicocele have qualitative, quantitative and functional abnormalities that may affect spontaneous fertilization. / Doutorado / Ciencias Biomedicas / Tocoginecologia

This dissertation reports on an investigation into relationships between human embryo, oocyte and follicular features and the developmental potential of the associated IVF embryos on transfer. An improved understanding could yield insights into biological factors influencing the development process. Besides, identifying the features associated with a good developmental potential could assist embryologists in an informed selection of the 'best' embryos for transfer. Previous studies with the same broad goal have suffered from a number of limitations. The present study attempts to overcome some of these by using the technique of 'decision tree' analysis developed in Machine Learning in Computer Science. The technique analyses data related to a sufficient number of specific examples in a group and discovers the general combinations of features characterising the group, presenting results in an easily comprehensible form. The feasibility of using the technique was first explored in a pilot study. The patterns of features characterising the 'good' embryos that resulted in triplet pregnancies on 3 embryo transfer or high hCG secretion in vitro and the 'bad' embryos that did not progress to the blastocyst stage on in vitro culture were identified. Given the promising nature of the results, a larger study, which forms the core of the dissertation, was conducted. Data included 53 features relating to the follicle, oocyte and embryo for batches of 3 embryos transferred that resulted in either a Take home baby (TH) outcome or a negative pregnancy test (NTH). The relationship between the 53 features and the 'TH' or 'NTH' developmental potential was investigated. The results identify, amongst others, two features related to the follicular stage of development as being important determinants of the TH potential, thereby confirming that developmental potential of an embryo is determined as early as the stage of the unfertilised oocyte. The embryo grading used by embryologists in the Oxford IVF Unit was identified as important, providing for the first time an unbiased corroboration of the selection process used in the Unit. The results refine the selection criteria and suggest two additional features, namely, follicular fluid volume and rate of cleavage as being worthy of further investigation, with the latter feature being indicated as the main component of the embryo grade characterising batches that resulted in a TH outcome. The relative unimportance of a number of features recorded in the patient notes but showing no relationship to development too is identified. The method of analysis used here could have wide-ranging applicability to a number of areas within IVF. A demonstration of this is provided in this dissertation in an area of growing importance, namely, the identification of blastocysts of different quality. The main study reported here can be extended to incorporate maternal and sperm features. Results obtained could form the basis for assisting embryologists in the selection of embryos with the 'best' developmental potential, with the possibility of an increase in IVF success rates.

M. Tech. / Infertility is defined as the inability to conceive after a minimum of one year of regular intercourse without contraception (Carlson et al, 2002). This may occur as primary infertility, where individuals have never had a biological child, or secondary infertility where individuals have had at least one previous documented conception (Greer et al, 2003). Infertility, in the African setting, is seen as a violation of the social norm. It contributes to psychological distress and marital instability as well as the loss of social security, social status and gender identity. Parenthood is considered culturally mandatory making childlessness unacceptable. Not only does Africa have the highest fertility rates in the world, Africa also has the highest number of infertility cases globally (Dyer et al, 2005; Ragone & Twine, 2000). Unexplained infertility is diagnosed when the routine investigation of semen analysis, tubal patency and assessment of ovulation show no abnormality and the couple have engaged in regular sexual intercourse. Unexplained infertility is thus a diagnosis of exclusion. When a previously identified cause of infertility is corrected, yet infertility persists a diagnosis of unexplained infertility is also given (Behrman et al, 1988). Before a diagnosis of unexplained infertility can be made four aspects are generally assessed. Each aspect encompasses a different aspect of the reproductive process: the number and quality of sperm, maturation and release of the ova, barriers to fertilization and barriers to implantation and maintenance of pregnancy (Cooper-Hilbert, 1999). The aim of this research was to evaluate the efficacy of homoeopathic similimum treatment in females pre-diagnosed with unexplained infertility using case studies. Cases were evaluated using fertility and parameters related to fertility which include basal body temperatures and midcycle cervical mucous changes. In addition, general parameters such as general well-being, premenstrual symptoms, dysmenorrhoea and sexual function were also evaluated. Concomitant symptoms were included in the analysis of each participant’s holistic case. The male partner was also required to be free of any structural and functional pathology.

Magister Psychologiae - MPsych / We live in a society that continually reinforces the connection between femininity and maternity and for the majority of women, attempts to experience motherhood are successful. However, for others the world of motherhood is not so easy to enter. To date, research regarding the needs and life satisfaction of women who are unsuccessful in becoming mothers, is fairly limited. The purpose of this study was to explore the transitional phase women endure from potential motherhood to non-motherhood and to highlight some of the complexities underpinning infertility and its impact on the lives of women in South Africa. The main objective was to gain deeper insight into how women incorporated this experience into their lives and relationships and how they began to create a future life without their own biological children. A secondary aim of this study was to investigate whether the women viewed aspects such as age and finances as having had an effect on their decision to discontinue treatment for infertility. Feminist standpoint theory served as a theoretical framework for the study that recognized that each individual voice be heard. This study was a qualitative exploration, utilizing a short demographic questionnaire and an in-depth semi-structured interview. Five interviews were conducted with women from diverse backgrounds. These interviews were recorded; transcribed verbatim and thematic analysis of the data was conducted. / South Africa

Approximately 1% of women will experience menopause before the age of 40, a condition known as premature ovarian failure (POF). The goal of this thesis was to identify genetic causes of POF by examining a number of candidate factors in POF patients and control women. Carriers of FMR1 premutations (55-200 CGG repeats) are known to be at increased risk of POF. A higher prevalence of alleles between 35-54 repeats was found among POF patients (p=0.01), suggesting that risk for POF may extend outside the classic premutation range. There was no evidence for any difference in FMRI promoter methylation or gene expression between cases and controls. Allele distributions of gene polymorphisms in the androgen receptor (AR), estrogen receptor (ß, sex hormone binding globulin, and FSH receptor genes did not differ between POF patients and controls. However, haplotype at the estrogen receptor a gene, ESRI, was found to be associated with POF in a simple dominant manner (RR=9.7; 95% CI=2.6-35.6). Although the functional effect of this haplotype could not be confirmed, it may confer a more active promoter that influences risk by increasing the rate of follicular atresia. X-chromosome inactivation (XCI) skewing can indicate an abnormal X chromosome and may therefore be increased in POF patients. There was no increase in skewed XCI >90% in patients with secondary amenorrhea, however, there was a significant increase in 4 POF patients with primary amenorrhea (p=0.001). No X-chromosome abnormalities were detectable by high resolution DNA microarray, and skewed XCI may be explained by a trisomic rescue event causing reduced follicular pool. Age-related chromosome factors were assessed to determine if POF patients demonstrate an increased rate of cellular aging. With age, XCI skewing and AR methylation increase and telomere length decreases. There was no difference in skewing or methylation between patients and controls. Surprisingly telomere length was increased in POF patients (p=0.04), a finding that may be explained by abnormal estrogen exposure. Genotype at the longevity-associated APOE gene was not associated with POF. In conclusion, these findings have illuminated several new areas of research in this field and provide background for future research into POF pathogenesis. / Medicine, Faculty of / Medical Genetics, Department of / Graduate

Heterochromatin can be described cytologically as those chromosomes or parts of chromosomes which remain heteropycnotic, or dark staining, through most of the cell cycle. Genetically and biochemically heterochromatic regions generally seem to be inert and it has been suggested that many heterochromatic loci are duplicated several times. In micro-organisms, genetic and biochemical analyses have been greatly facilitated by the use of conditional lethals which survive under "permissive" conditions but die under "restrictive" conditions. Temperature-sensitive ethyl methanesulfonate-induced lethal mutations (such mutants result in survival at 22°C but death at 29°C) have previously been used in Drosophila melanogaster for preliminary studies of development. In the present study 8 temperature-sensitive (ts) sterile mutations (males are fertile at 22°C but sterile at 29°C) were induced on the Y chromosome of D. melanogaster. The ts mutants were mapped genetically on the long arm of the Y chromosome and they were found to involve a minimum of 4 different loci. The Y chromosome of D. melanogaster is entirely heterochromatic and it is necessary for male fertility but the exact function of the Y chromosome is uncertain. The recovery of point mutations (ethyl methanesulfonate-induced temperature-sensitive mutations are presumed to be point mutations) on the Y chromosome indicates that there are loci on the Y represented by a single copy. A determination of the specific developmental effects of the ts sterile mutations, was also attempted. By exposing mutant males to a 48 hour period under the restrictive conditions (29°C) and observing their fertility for several days, the stage in the production of mature sperm during which the ts mutants were having an effect, was determined. / Science, Faculty of / Zoology, Department of / Graduate

Includes bibliographical references (leaves 286-297). / In the communities of the Cape Flats, it is expected that all women will bear children and become mothers. Motherhood serves as a social and cultural indicator of femininity and enables women to access social and economic networks that knit them into community. The social and cultural valorization of motherhood in these communities has informed the powerful stigmatization of infertility (or the involuntary nonconformance to motherhood). The stigma associated with infertility affects women in particular, because the inability to bear children is commonly perceived to be a woman's problem. This study explores the cultural constructions of infertility. It examines in particular, the diverse cultural meanings and the stigma associated with infertility. The examination of these cultural meanings challenges the notion that infertility should only be examined in the biomedical realm. My research was conducted over a seven month period with six infertile women and with women who have borne children from different areas on the Cape Flats. The infertile women were the primary informants. Other informants included the mothers with whom the focus group was conducted and specialist informants who were healthcare professionals. The participants were recruited through the primary health care clinic in Manenberg, the network of community newspapers, The Daily Voice and through my own social network. Qualitative research methods were used. The study also used participatory research methods involved because the participants played an active role in the construction of the research process and interview schedules. The primary information used was obtained from in-depth interviews and journals kept by the infertile women. For comparative purposes, a focus group was conducted with a group of mothers. The study illustrates that on the Cape Flats, infertility is constructed as a major cultural and social problem for women. The stigma attached to infertility draws its power from the social and cultural meanings associated with inability of infertile women to live up to the expectation that every adult woman will become a mother. The effects of the social stigma of infertility are especially profound. As I show, bio-medicine does offer some solution, but only to the few who can afford it.

Si la fécondation in vitro est une innovation biomédicale récente, datant seulement de 1978, le recours à l’Assistance Médicale à la Procréation (AMP) est rapidement entré dans les mœurs, au point que ses procédés participent aujourd’hui à plus de 3 % des naissances françaises. Les cliniciens et les biologistes de la reproduction jouent un rôle incontournable dans ce champ. Chefs d’orchestre de la rencontre des gamètes, ils sont également les principaux acteurs décisionnaires de l’accès et de la poursuite des traitements de l’infertilité. Pourtant, leur pouvoir décisionnel a moins été étudié que les conséquences de leur intervention en termes de vécu de l’infertilité, de parenté, de filiation et de genre. C’est précisément ce manque que cette recherche tente de pallier. Au croisement des sociologies de la santé, des professions, des organisations et de l’action, cette étude vise à éclairer la multiplicité des pratiques et des représentations des spécialistes de l’infertilité à travers le prisme des décisions médicales. Elle repose sur deux enquêtes, menées pendant plus de deux ans au sein de plusieurs centres d’AMP de la région parisienne, comprenant des observations de consultations et de réunions pluridisciplinaires, et des entretiens avec des patients et des spécialistes de l’infertilité. S’appuyant sur ce corpus de données, et guidée par un souci constant d’articuler les dimensions micro et macro sociologiques, cette investigation s’attache d’abord à resituer les contextes globaux et locaux dans lesquels s’inscrivent les choix des professionnels, puis à analyser les décisions relatives à l’accès à l’AMP et aux techniques médicales. In fine, cette recherche montre que la sélection des patients et les stratégies médicales, structurées dans des cultures propres à chaque centre, apparaissent comme des compromis trouvés par les acteurs qui composent avec un ensemble de contraintes pour adopter des pratiques en adéquation avec leurs valeurs et leurs représentations. / The in vitro fertilization is a recent biomedical innovation, dating from 1978. Nevertheless, the use of Assisted Reproductive Technologies (ARTs) become widely accepted, to the point where these technics contribute today to more than three per cent of French childbirth. Clinical practitioners and biologists play a key-role in this field. They are in charge of the gametes meeting and they are also the main decision makers on the access and the pursuit of infertility treatments. However, their decisional power has been less studied than the consequences of their intervention in terms of infertility experience, kinship, filiation and gender. This is precisely what this research aims to overcome. At the crossroads of health sociology, sociology of professions, of organizations and of action, the goal of this study is to highlight, through the prism of medical decisions, the multiplicity of practices and the diversity of representations of infertility specialists.This research is build on a two years of qualitative study lead in several ART centres in the Parisian region, including observations of consultations, multidisciplinary staff meetings, and interviews with patients and infertility specialists. Based on this data, and constantly trying to articulate micro and macro sociological dimensions, this investigation is firstly devoted to report the local and global contexts where the professionals’ choices are taken, and then to analyse decisions about ARTs access and medical technics.In fine, this research points out that the patients’ selection and the medical strategies, each different depending on the specific ART centers’ cultures, appear as a compromise made between the different actors, who have to compose with a set of constraints to, in the end, start off practices in adequacy with their own values and representations.

Antisperm antibodies (ASA) are thought to be a predominate cause of immune infertility by interfering with various aspects of sperm function in both the male and the female reproductive tracts. The precise mechanism by which these antibodies contribute to infertility, as well as their etiology, remains to be established. ASA are present in a variety of biological substrates, such as genital tract secretions, and the blood sera of both males and females. Although not all ASA underly infertility, a substantial body of research suggests that certain ASA, referred to as sperm immobilizing antibodies (SI-Abs) and sperm agglutinating antibodies, significantly impair sperm transportation in the female reproductive tract. High titers of sperm agglutinating or sperm immobilizing antibodies have been associated with reproductive failure. CD52g is a GPI anchored glycoprotein found on mature sperm and in seminal plasma (SP). Antibodies against a male reproductive tract-specific epitope of CD52g are known to readily agglutinate sperm. The current study sought to develop an ELISA to quantify the prevalence of CD52g antibodies in the sera of male and female patients with infertility, and to determine if there was a correlation between the prevalence of CD52g antibodies and the prevalence of sperm agglutinating antibodies in the sera of these patients. Ultimately, CD52g antibodies were only detected in the sera of patients (21%) with sperm agglutinating antibodies. While detecting CD52g antibodies in sera via an ELISA proved challenging, the results of this study corroborate research demonstrating that CD52g antibodies have a remarkable capacity to agglutinate sperm. Elucidation of the mechanisms underlying this immune response would advance our understanding of immune modulation in human reproductive tracts, further the diagnosis of immune infertility, and are currently providing the basis for the development of a potent dual purpose immunocontraceptive, that both prevents unintended pregnancy, and prevents the transmission of sexually transmitted infections (STIs).