Bacterial contamination of the female genital tract after parturition is common and subsequent infections are associated with infertility. Infections of the uterus or mammary gland in cattle perturb ovarian function, with decreased dominant follicle growth rate and delayed ovulation. Granulosa cells that line ovarian follicles express the molecular machinery necessary to respond to common bacterial pathogen-associated molecular patterns (PAMPs), suggesting that granulosa cells may play a role in ovarian innate immunity. In the present thesis, the effect of bacterial PAMPs on ovarian follicle cells was examined during follicular growth, from emergence through dominance, and the early stages of embryogenesis. Granulosa cells from emerged or dominant bovine follicles increased production of inflammatory mediators, such as IL-6 and IL-8, through Toll-like receptor (TLR)-dependent pathways in response to the PAMPs lipopolysaccharide and the synthetic triacylated lipopeptide Pam3CSK4. The requirement for TLR2 and TLR4 was confirmed by using siRNA, showing activation of MAPK intracellular pathways and inhibiting key signals (p38, MEK or NFkB) in TLR pathways. The endocrine function of granulosa cells was perturbed by PAMPs, with decreased oestradiol and progesterone output. In addition, the endocrine environment affected granulosa cellular responses, with high EGF increasing the cellular response to PAMPs. Bacterial PAMPs also perturbed the oocyte, with increased cumulus expansion and parthenote cleavage rate, as well as changes in key genes involved in oocyte maturation. Finally, a human granulosa cell line was used to examine if PAMPs perturb human as well as bovine ovarian health. Indeed, human cells also expressed TLRs and mounted a cellular response to PAMPs at the mRNA level. This thesis provides a molecular mechanism for the perturbation of ovarian function by an infection at a site distal to the ovary.

High-grade serous ovarian carcinoma (HGSOC) is characterised by genomic instability, ubiquitous TP53 loss, widespread disease at diagnosis and the frequent emergence of platinum resistance. This thesis explores the use of high-throughput genomics technologies to understand if resistance could be explained by the model of tumour evolution. We performed SNP array analysis of a cell line model system of platinum resistance consisting of matched cell lines from three cases of HGSOC established before and after clinical resistance developed, the OVOl clinical study consisting of six matched pairs of tumours before and after three cycles of chemotherapy, and the OV03/0V04 study consisting of 18 cases sampled at multiple timepoints and from multiple metastatic sites. The results showed evidence of metastatic site dependent divergence. Moreover, mutually exclusive loss of heterozygosity patterns between presentation and relapse genomes, including all the cases in the cell line system and one of two OV03 cases for which relapse material was available, suggest that the relapse arises from a minor subclone of the presentation disease, while in the remaining case, the subclone with an NFJ homozygous deletion was enriched in the relapsed disease. I then asked which mutational process drives evolution. Using next-generation sequencing (NGS), I compared the structural variants between and within cases in the model system and in 6 cases of the OV03 cohort. From the genomic signatures in the cell lines, I demonstrated that a case with homologous recombination (HR) deficiency acquired numerous translocations and small deletions (median size of 13.4kb) , whereas another showed a novel tandem duplicator phenotype (median size of tandem duplications was 350kb). Mutator phenotypes in both cases arose early in progression and persisted, but the tumour with HR deficiency showed evidence of re-stabilising its ,"genome and lost platinum sensitivity after a revertant BRCA2 mutation restored its HR function. A subset of tumours from the Cancer Genome Atlas (TCGA) dataset suggested that these two phenotypes were mutually exclusive. Amongst the six OV03 cases, preliminary analysis suggests that one case showed an amplifier phenotype and three cases showed evidence of parallel evolution. Taken together, early onset of mutator phenotypes and parallel evolution may provide a mechanism by which resistance evolves. Further work should aim to identify the processes involved in tumour evolution in 'purified' populations such as cancer stem cells.

Ovarian changes, patterns of reproduction, and age-specific productivity of female Columbian black-tailed deer (Odocoileus hemionus columbianus) on Vancouver Island were elucidated largely by examination of serial, stained sections of ovaries from 444 females. Well-defined, 8 or 9-day, follicular cycles occur during the breeding season in November and early December. Many follicles rupturing at first ovulation are asynchronous and of extreme sizes. Relatively small (4-5 mm³), short-lived (8-day) corpora lutea develop after first ovulation; these rapidly regress after second ovulation, while the corpora lutea of pregnancy grow to 100 mm³, within 5-8 days. The changes in ovarian structures during the breeding season were ascertained in 12 females, whose follicles and corpora lutea were dated by the cleavage stage of fertilized ova. These provided criteria for estimating ovulation dates in other females. In each cycle, about 50% of the adult females ovulated within an 8-day period. The mean date of first ovulation in each of the 5 years was approximately November 16; the second ovulation followed after a remarkably short period of 8-9 days. About 96% of the females conceived at second ovulation and 4% on subsequent ovulations. A high proportion of 'silent heats' accompany first ovulation, as indicated by lack of sperm on four of six ova. Growth curves, based on fetuses conceived at second ovulation, provided the conception dates of two late conceivers—those containing more than one generation of regressed corpora lutea of non-pregnancy. Females may cycle at least five times if pregnancy does not occur. Accessory corpora lutea develop in large and small unruptured follicles, in small ruptured follicles, and in regressing corpora lutea. They occurred in 47% of females between first and second ovulation and in 36% of pregnant females. Minimum loss of ova was 8.3% in all females that produced corpora lutea and 4.3% in females that became pregnant, of which 3.1% was moribund fetuses. Corpora lutea of pregnancy, corresponding to the number of fetuses, develop into distinctive scars, which persist for the life of the doe. Pregnancy rates of the previous season, as well as the long term productivity of individuals and the population, were estimated from these. The average number of viable fetuses per doe increased progressively from 0.91 in yearlings to 1.81 in the 5-5 to 6.5 age-classes, and thereafter decreased. These changes in fertility with age were mirrored by changes in weight and girth. The relative contribution of fawns by each age-class in the population progressively decreased with age. One hundred females of reproductive age produced about 137 fawns. / Science, Faculty of / Zoology, Department of / Graduate

No description available.

LD2668 .T4 1962 L44

published_or_final_version / abstract / Clinical Oncology / Doctoral / Doctor of Philosophy

published_or_final_version / abstract / Pathology / Doctoral / Doctor of Philosophy

published_or_final_version / abstract / Biological Sciences / Doctoral / Doctor of Philosophy

published_or_final_version / abstract / Biological Sciences / Master / Master of Philosophy

published_or_final_version / Pathology / Master / Master of Philosophy