Objective: To determine whether serum phthalate concentrations differ in men and women with infertility compared to those without infertility in Saskatchewan Hypothesis: Serum phthalate concentrations will be greater in men and women with infertility compared to fertile men and women Setting: Patients undergoing assisted reproduction for the treatment of infertility; healthy volunteers recruited from the community Recruitment and sample collection: Infertile couples were recruited prior to in vitro fertilization (IVF) therapy for treatment of unexplained infertility (n=15), polycystic ovarian syndrome (PCOS, n=13), and male factor infertility (n=12); fertile men (n=15) and women (n=15) were recruited using poster advertisements. Blood samples were collected by venipuncture for phthalate analysis. Main outcome measures: Serum phthalates concentrations (ng/mL) Design: Prospective cohort pilot study Methods: In infertile couples, blood samples were collected on the following 3 days of the IVF cycle: early during ovarian stimulation, day of oocyte retrieval and day of embryo transfer. In healthy volunteers, 3 blood samples were collected over a 2 week period. Liquid chromatography and tandem mass spectrometry (LC-MS/MS) was conducted to quantify concentrations of four phthalates: di-n-butyl phthalate (DBP), diethyl phthalate (DEP), di-(2-ethylhexyl) phthalate (DEHP) and diisodecyl phthalate (DIDP). Phthalate concentrations were compared among the four study groups using non-parametric Kruskal-Wallis and Mann-Whitney U post hoc tests. Results: Serum DEHP and DEP concentrations did not differ among control, unexplained, PCOS, and male factor infertility groups in both men and women (p>0.05). DBP in women did not differ among study groups (p=0.205). In contrast, DBP was lesser in men with unexplained, PCOS, and male factor infertility compared to controls (p < 0.05). Similarly, DIDP was lesser in women of couples with unexplained, PCOS and male factor infertility groups compared to fertile women (p < 0.05). Less DIDP was detected in men with unexplained and male factor infertility compared to the control group (p < 0.05) Conclusion: Serum phthalate concentrations in serum were lesser or not different in infertility patients undergoing IVF compared to fertile volunteers. These findings do not support the notion that serum phthalate concentrations are associated with human infertility. Further research is needed to determine whether phthalate concentration in blood cells and adipose tissue differ in infertile versus fertile men and women.

Introduction: In recent years, the average age of first reproduction has risen significantly, the mean now standing at around 30 years in many countries. The adverse effects of maternal age on fertility and reproduction have been well documented. However, the influence of paternal age on fertility, reproduction and postnatal health is relatively poorly understood, and 50% of all male infertility cases are classed as idiopathic or unexplained infertility. Methods: The aim of this study was to investigate factors that contribute to male infertility, split into two main parts. The first part focused on analysing data collected from patients who had undergone fertility treatment to assess the influence of different factors on infertility, especially at the genome level. The second part attempted to deal with some of the technical challenges of screening and diagnostic methods to study the genome, with the aim of providing tools that would assist future studies in pinpointing genetic factors responsible for infertility, especially in cases of idiopathic infertility. Results: Based on data from the first part of the study, it was determined that advanced paternal age can affect sperm progressive motility, sperm DNA integrity and the fertilisation rate of in vitro fertilisation (IVF) cycles, as well as the development of embryos. Direct analysis of sperm DNA fragmentation (SDF) and degradation levels revealed an association between elevated SDF and impaired embryo development. Furthermore, a correlation was shown between chromosome aneuploidy and variance in SDF and sperm DNA degradation. Moreover, aneuploidy can influence abnormal sperm morphology and consequently also progressive motility. Also, embryo development rate of IVF cycles on day three, demonstrated a significant decline in cycles where the sperm used for fertilisation had a high aneuploidy rate, which can highlight the reduced developmental capacity of aneuploid embryos. From the lifestyle factors assessed, only alcohol consumption significantly correlated with the sperm DNA damage. Therefore, poor semen quality may highlight damage that has been incurred by the sperm DNA. When the semen quality is suboptimal, the intracytoplasmic sperm injection (ICSI) technique is suggested as a standard strategy to improve the prognosis of ART. However, when the progressive motility is poor, the ICSI approach is not as effective. Based on our findings and in line with other studies, the only sperm parameter that can be affected by paternal age is sperm motility, which could be an indicator of SDF. Therefore, the decline in ICSI fertilisation rate in patients with impaired sperm progressive motility could be due to sperm DNA damage, and even ICSI cannot improve the fertilisation rate considerably. Discussion: The aim of the second part of this project was to establish a robust workflow for whole- genome amplification (WGA) and whole-genome sequencing of single cells to improve the coverage rate and fidelity, with the aim of providing means of detecting any mutation in the genome that might be responsible for reduced embryonic developmental competence. Towards this end, the efficiencies of two different WGA protocols (REPLI-g and TruePrime) were compared. Multiple technical factors required optimisation in order to create a suitable protocol. Our results demonstrated the overall superiority of REPLI-g compared to TruePrime in almost all the assessed parameters. The amplification rate of REPLI-g was much faster than that of TruePrime, and prolonged incubation led to overamplification and an increased duplication rate. However, the TruePrime method has a slower amplification rate and therefore, by increasing the incubation time, it was possible to improve the quality of the data. The modified protocol with reduced volume also had the most promising outcome in terms of the data produced, and could fulfil our expectations by being fast, cost-effective and efficient. Conclusion: In conclusion, the results from the first part of this study confirmed the negative impact of male age on assisted reproductive treatments, which can result in decreased success rates of fertilisation. Other factors such as sperm DNA damage may also contribute to this age effect, suggesting that assessing this parameter prior to fertility treatment, and attempting to mitigate elevated levels of sperm DNA damage, may be of value to older patients. Additionally, overcoming the technical challenges in studying genetic contributory factors in infertility is a promising step toward better understanding of the mutations and variations that are involved in this phenomenon.

In couples diagnosed with unexplained infertility, the woman has normal ovulatory cycles and no organic pathology, and the man has normal semen. The period during which the implantation of an embryo can occur is defined as the window of implantation (WOI). For infertile women, disturbances in the endometrium maturation are hypothesized to impair the uterine receptivity for embryo implantaition. However, there are still no methods found to predict endometrial receptivity. Hemoglobin related-proteins hypothesized to be involved in embryo implantation have been found in the endometrium in both fertile and infertile women. The aim of this study was to determine whether there were differences in the expression of hemoglobin-related proteins, hemoglobin-α, cytoglobin, fetal hemoglobin and haptoglobin in the endometrium of fertile and infertile women. Immunohistchemical staining was used. Haptoglobin showed negative expression in glands, luminal epithelium and strong expression in stroma for both gruops. The expression of the hemoglobin-related proteins, hemoglobin-α, cytoglobin, fetal hemoglobin intensity were stronger in fertile than infertile women. More studies are required to find other factors that may have an effect on fertility.

Submitted by Renata Lopes (renatasil82@gmail.com) on 2016-01-12T17:34:43Z No. of bitstreams: 1 nataliamachadobrito.pdf: 1118656 bytes, checksum: 57ac8f9e9647b471a806d0e2ac6b603f (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2016-01-25T17:23:39Z (GMT) No. of bitstreams: 1 nataliamachadobrito.pdf: 1118656 bytes, checksum: 57ac8f9e9647b471a806d0e2ac6b603f (MD5) / Made available in DSpace on 2016-01-25T17:23:39Z (GMT). No. of bitstreams: 1 nataliamachadobrito.pdf: 1118656 bytes, checksum: 57ac8f9e9647b471a806d0e2ac6b603f (MD5) Previous issue date: 2015-02-27 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A teoria do Medo (Angst) de Freud postula que o ego, ao perceber a ameaça de desejos inconscientes, desencadeia o afeto de medo, que por sua vez aciona o recalque, podendo, então, desencadear a formação de sintomas neuróticos, como também os somáticos. Aqui, o medo neurótico, da mesma forma que uma grave ameaça real, mantém o sistema fisiológico em funcionamento alterado caracterizando o processo de estresse crônico, que desencadeia as doenças psicossomáticas. A partir dessa premissa, suspeita-se a ISCA possa ser outro sintoma. Esta pesquisa qualitativa tem como objetivo pesquisar uma possível relação entre medo e ISCA, analisando possíveis vivências estressantes e às manifestações clínicas do medo em mulheres que fazem tratamento. A coleta de dados se deu por meio de entrevistas semiestruturada e de um diário de campo. Para análise dos dados adotou-se a Análise de Conteúdo, em específico a análise temática. A análise dos resultados evidenciou eventos estressores, anterior à decisão de engravidar e na fase do tratamento. Tais eventos foram acompanhados de sintomas somáticos e neuróticos. Pode-se inferir que a ISCA talvez possa ser uma consequência do processo de defesa psíquica, embora não seja possível desvendar este enigma, pois teríamos que entrar numa discussão de causalidade, que na verdade não diz nada quando estamos diante do inconsciente. / The Unexplained Infertility (UI) is a medical diagnosis given when there is no identified changes signals related to reproduction. But what escape of scientific knowledge when we cannot identify the infertility`s cause? This research bet that there is something of the order of fear and psychic defense that deserves more attention, since the point of view of the unconscious productions, UI might be another symptom. This qualitative research aims to investigate the relationship between fear and UI, analyzing possible stressful experiences and clinical manifestations of fear in women under treatment. The data collection was carried out through interviews semi-structured interviews and a diary. For data analysis we adopted the Content Analysis, specifically thematic analysis. The results showed stressful events preceding the decision to get pregnant and at the stage of treatment. These events were followed by somatic and neurotic symptoms. It can be inferred that the UI might be a consequence of psychic defense process, although it is not possible to unravel this puzzle, because we would have to enter into a discussion of causality, which actually says nothing when we face the unconscious.

Thesis advisor: Barbara E. Wolfe / Studies suggest that eating disorder (ED) pathology may be linked to ovulatory and unexplained infertility in women who present to reproductive treatment centers. Specifically, studies have linked hypothalamic amenorrhea, oligomenorrhea, and anovulatory cycles to disordered eating. Advances in Assisted Reproductive Technology can lead to successful conception for women with ED; however, they have a higher risk for poor maternal and fetal outcomes. This descriptive, comparative, quantitative study examined disordered eating and psychological symptoms in women with ovulatory and unexplained infertility compared with women receiving routine health care from their primary care providers. Women ages 20 to 44 were recruited. After providing verbal consent, a study packet was mailed to the study participant's home. The Eating Disorder Inventory-3-Referral Form and Herman and Polivy Restraint Scale measured disordered eating symptoms. The Speilberger Anxiety Inventory and Beck Depression Inventory-II examined psychological symptoms. Provisional DSM IV TR diagnoses were ascertained using the Eating Disorder Inventory-3-Symptom Checklist, along with other scale items. Multivariate analysis of covariance (MANCOVA) confirmed that women with ovulatory and unexplained infertility had significantly higher Desire for Thinness (p = .001) and Bulimia (p = .007) subscale scores putting them at risk for Anorexia Nervosa or Bulimia. Women receiving routine care had significantly higher Body Dissatisfaction (p = .000) subscale scores consistent with their higher weight and tendency toward overeating. Women receiving routine care also had significantly higher Restraint (p = .000) scale scores, leaving them at risk for dietary disinhibition. The groups did not differ on psychological symptoms. Women with infertility had lifetime ED diagnoses many times the national ED prevalence rate, similar to the research findings of Freizinger et al. (2010). The study results support that women with ovulatory and unexplained infertility are at risk for having an occult ED. The critical import of integrating ED assessment into infertility evaluation, reproductive and primary care was implicated. Further study to isolate biobehavioral markers to better identify women at risk for ED and improve their maternal and fetal outcomes was recommended. / Thesis (PhD) — Boston College, 2010. / Submitted to: Boston College. Connell School of Nursing. / Discipline: Nursing.

Infertility is defined as not being able to conceive after 1 year of regular intercourse without use of contraception. Unexplained infertility is a diagnosis given to couples where the reason to infertility cannot be clarified even after the routine examination. Undefined infertility is a common and growing problem because most people are not aware of the fact that fertility decreases after the age of 35. Hyper- and hypothyroidism has been known to affect the menstrual cycle as well as increased risk of miscarriage. However, the specific effect of thyroid hormones on infertility has not yet been clarified. This study aims to compare the gene expression of two thyroid hormone receptors DIO2 and MOT8 in human endometrium and fallopian tube tissue from two phases of the menstruation cycle, follicular phase and lutheal phase. The methods used were RT-qPCR and immunohistochemistry, which showed a statistically significant difference in the expression of DIO2 and MOT8 between fallopian tube tissue and endometrium, but not between follicular and lutheal phase. However, MOT8 seemed to have a tendency to be down-regulated in the follicular phase but the results need to be validated with different endogenous controls and larger study groups.