INTRODUCTION Phthalates, a chemical class of plasticizers, are ubiquitous in the environment and recognized as endocrine disrupting compounds (EDCs). Recent data suggest that oxidative stress is a potential mediator of poor male reproductive health associated with phthalate exposure. Mitochondria are implicated in the production of excess oxidative stress and sperm mitochondrial copy number (MtCopy) and deletions (MtDeletion) have been linked with male infertility. However, little is known about the relationship of these mitochondrial biomarkers in sperm with phthalate exposure and oxidative stress. OBJECTIVES To examine associations of urinary phthalate metabolites and isoprostane concentrations on sperm MtCopy and MtDeletions in male partners undergoing assisted reproductive technologies (ART). METHODS A total of (n=97) sperm samples were collected from male partners undergoing ART at Baystate Medical Center, in Springfield, MA from 2014 to 2016 as part of the Sperm Environmental Epigenetics and Development Study (SEEDS). Seventeen urinary phthalate metabolites (n=103) were analyzed by the Centers for Disease Control using tandem mass spectrometry. 15-F2t-Isoprostane (n=101) was measured using a competitive enzyme-linked immonsorbent assay in urine of male individuals. A triplex Taqman probe-based qPCR method was developed for relative quantification of genomic DNA, MtCopy and MtDeletions. Multivariable linear or logistic regression was employed to examine associations with age, BMI, batch and current smoking status with each outcome to determine confounders used for adjustment. RESULTS Quartiles of MtCopy and MtDeletion were positively associated with the odds of male infertility (p for trend < .0001 and 0.007, respectively). Urinary metabolite concentrations of MCNP displayed a positive association with MtCopy (β=1.56; p =0.03). Urinary MEHP concentrations were positively associated with MtDeletion in only infertile individuals (n=30) (β = 0.075; p = 0.006). Urinary isoprostane concentration was not associated with MtCopy or MtDeletion, but was associated with seven phthalate metabolite concentrations (MEOHP, MEHHP, MBzP, MHBP, MiBP, and MHiBP). CONCLUSIONS To our knowledge, this is the first study to investigate the relationship between sperm MtCopy and MtDeletion with oxidative stress and phthalates. These results suggest that certain phthalate metabolites may be associated with a known biomarker of systemic oxidative stress. Sperm mitochondrial function as measured by MtCopy and MtDeletion may be considered biomarkers of male infertility, although no relationship was shown between mitochondrial outcomes and oxidative stress. Future research is investigating these relationships with developmental outcomes including embryo quality.

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In the United States, 6.7 million or 10.9% of women, between the ages of 15 and 44 have difficulty becoming or staying pregnant and of these women 6% are infertile (Centers for Disease Control and Prevention [CDC], 2012). Infertility is defined as twelve months of regular, unprotected intercourse yielding no pregnancy (Tierney, McPhee, & Papadakis, 1999). For those struggling with infertility there can be numerous negative psychological problems, including depression and anxiety (Jordan & Revenson, 1999). Given the widespread and negative impact of infertility, it is important to further understand and explain the experiences of infertile women. The present work contributes to the literature by qualitatively examining women’s infertility as a stigmatizing experience. Goffman (1963) defines stigma as an attribute that society deems as deeply discrediting and reduces an “individual from a whole and usual person to a tainted, discounted one” (p. 3). Moreover, individuals who possess a stigmatizing condition are likely viewed as a less or inferior person. Because women struggling with infertility are unable to fulfill societal norms that mandate motherhood (e.g., Gonzalez, 2000; Whiteford & Gonzalez, 1995), women experiencing infertility may perceive stigma. The aim of the current study is to explore infertility as stigmatizing in order to provide a more complete framework for understanding negative psychological outcomes among women with infertility. To this end, we conducted qualitative interviews with nine women who had been trying to conceive without success for at least one year. Participants were recruited by newspaper advertisements, campus emails, flyers in local nurse practitioner clinics and local infertility clinics in Johnson City, TN. Interviews were conducted one-on-one and were guided by general questions about the most difficult aspects of infertility and how women’s lives, including social relationships and beliefs about themselves, had been impacted by the experience. Audio-recorded interviews were transcribed verbatim and coded for themes. Five reoccurring themes were identified: culture and norms, feelings of failure and loss of 2013 Appalachian Student Research Forum Page 115 womanhood, infertility as stigmatizing condition, change of world view, and social support. Each of these themes can be discussed in relation to the public and self aspects of stigma. For example, the expectation that females progressing into adulthood will eventually have children influenced infertile women’s interactions with the world around them causing them to feel inadequate and question their purpose in life. Moreover, women in our study reported being constantly reminded of their infertility struggles through everyday social interactions with strangers, family, and friends, some of which included differential treatment due to infertility. These continual reminders led women to internalize the negative beliefs regarding not living up to societal gender expectations or stereotypes. Furthermore, women’s reports of alienation and self-isolation, as well as anger and frustration map onto previous models of stigma-related processes (e.g., Hatzenbuehler, 2009; Richman & Leary, 2009. Thus, stigma theory may provide a framework with which to more fully understand the negative psychosocial outcomes commonly reported among women encountering infertility.

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No / Modern reproductive technologies are enabling the treatment of infertile men with severe disturbances of spermatogenesis. The possibility of elevated frequencies of genetically and chromosomally defective sperm has become an issue of concern with the increased usage of ICSI, which can enable men with severely impaired sperm production to father children. Several papers have been published reporting aneuploidy in oligozoospermic patients, but relatively little is known about chromosome structural aberrations in the sperm of these patients. METHODS: We examined sperm from infertile, oligozoospermic individuals for structural and numerical chromosomal abnormalities using a multicolour ACM fluorescence in situ hybridization (FISH) assay that utilizes DNA probes specific for three regions of chromosome 1 to detect human sperm that carry numerical chromosomal abnormalities plus two categories of structural aberrations: duplications and deletions of 1pter and 1cen, and chromosomal breaks within the 1cen¿1q12 region. RESULTS: There was a significant increase in the average frequencies of sperm with duplications and deletions in the infertility patients compared with the healthy concurrent controls. There was also a significantly elevated level of breaks within the 1cen¿1q12 region. There was no evidence for an increase in chromosome 1 disomy, or in diploidy. CONCLUSIONS: Our data reveal that oligozoospermia is associated with chromosomal structural abnormalities, suggesting that oligozoospermic men carry a higher burden of transmissible, chromosome damage. The findings raise the possibility of elevated levels of transmissible chromosomal defects following ICSI treatment.

No description available.

Currently infertility affects 10 to 15 percent of couples. In nearly 50% of all cases male factor contributes to infertility of the couple. Majority of causes of male infertility remains unexplained. For this reason, finding simple and clinically useful tools for improving male infertility diagnostics can be important at present. Based on the analysis of sperm transcriptome, in this diploma thesis we aimed to find genes that show differential expression between normal and pathological sperm, this could provide information about molecular basis of male infertility, moreover, expression profile of such genes in sperm could be employed for noninvasive diagnosis of male infertility. This study was conducted by using 67 sperm samples, including 16 control sperm samples from fertile men. The samples of infertile patients were divided into 3 groups according to morphology and motility using results of standard sperm evaluation according to WHO. First group included 20 patients diagnosed with astenoteratozoospermia (low sperm motility and abnormal morphology), second group was 15 patients diagnosed with asthenozoospermia (normal morphology and low sperm motility), the third group comprised 16 samples from infertile patients with normal spermiogram. 16 control samples were from fertile men (conception of...

Currently infertility affects 10 to 15 percent of couples. In nearly 50% of all cases male factor contributes to infertility of the couple. Majority of causes of male infertility remains unexplained. For this reason, finding simple and clinically usefµl tools for improving male infertility diagnostics can be important at present. Based on the analysis of sperm transcriptome, in this diploma thesis we aimed to find genes that show differential expression between normal and pathological sperm, this could provide information about molecular basis of male infertility, moreover, expression profile of such genes in sperm could be employed for noninvasive diagnosis of male infertility. This study was conducted by using 67 sperm samples, including 16 control sperm samples from fertile men. The samples of infertile patients were divided into 3 groups according to morphology and motility using results of standard sperm evaluation according to WHO. First group included 20 patients diagnosed with astenoteratozoospermia (low sperm motility and abnormal morphology), second group was 15 patients diagnosed with asthenozoospermia (normal morphology and low sperm motility), the third group comprised 16 samples from infertile patients with normal spermiogram. 16 control samples were from fertile men (conception of...

Androgen receptor (AR) signalling is essential for the development and function of the male reproductive system. Conditional gene ablation using the Cre-loxP system has previously assisted in the elucidation of the role of AR in different cell types. The aim of this study was to examine the effects of the ablation of AR in previously untargeted cell types, with the hypothesis that this will have significant and novel effects on reproductive development and function that have not been previously documented by current models of androgen disruption. In these studies, three Cre recombinase lines were empirically validated for action in the male reproductive system, before being used to ablate AR and the phenotypes of the resulting lines were characterised. Endothelial-specific receptor tyrosine kinase (Tie2)-Cre was shown to target the vascular and endothelial cells of the testis, and used to ablate AR in these cells. The testes of the resulting Tie2-ARKO line were morphologically similar to controls, with normal spermatogenesis and mature spermatozoa present in the cauda epididymis. Aquaporin 2 (Aqp2)-Cre was shown to target the post-meiotic germ cells of the testis, and was used to ablate AR in these cells. The testes of the resulting Aqp2-ARKO line were morphologically similar to controls, with normal spermatogenesis and mature spermatozoa present in the cauda epididymis. It was concluded that the Ar gene was dispensable in the endothelial cells and post-meiotic germ cells of the testis for normal spermatogenesis. Forkhead box protein G1 (FoxG1)-Cre was shown to target the caput epididymal epithelium and pituitary, and used to ablate AR in these cells. d100 FoxG1-ARKO mice had a severe testicular phenotype, with sloughing of the seminiferous epithelium, atrophy of some seminiferous tubules and distension of the rete testis with spermatozoa. Despite the severe testis phenotype, ablation in the testis was incomplete and restricted to a small percentage of Leydig cells, with no ablation in Sertoli cells. Ablation of AR in the embryonic pituitary did not cause adult serum testosterone or LH concentrations to change, nor did it cause changes in other pituitary hormone transcripts. Mosaic ablation of AR in the caput epididymal epithelium was shown to impair epididymal development, with failure of initial segment (segment I) development and a significant decrease in epithelial cell height and lumen diameter in the remaining proximal caput epididymis (segment II). Dysfunction of the caput epididymis resulted in the failure of spermatozoa to transit the efferent ducts into the epididymis correctly: instead they were found to stall in the efferent ducts and produce a block. The testicular phenotype could be explained as the result of fluid backpressure effects resulting from the efferent duct block. Consequently, low concentration of spermatozoa in the cauda epididymis resulted in infertility in the FoxG1-ARKO, which represents a new model of obstructive azoospermia.

published_or_final_version / Zoology / Doctoral / Doctor of Philosophy