Source: Masters Abstracts International, Volume: 46-05, page: 2669. Adviser: Margaret Caughy. Includes bibliographical references.

Thesis (Ph.D.) - University of Queensland, 2003. / Includes bibliography.

Thesis (Ph.D. in Molecular Biology) -- University of Colorado at Denver and Health Sciences Center, 2005. / Typescript. Includes bibliographical references (leaves 165-185). Free to UCDHSC affiliates. Online version available via ProQuest Digital Dissertations;

Thesis (M.S.)--Indiana University, 2004. / Includes bibliographical references (leaves 67-88).

Increased HIV susceptibility has been associated with the progesterone-dominant luteal phase of the menstrual cycle and the use of progesterone-only contraceptives, yet the mechanism is poorly understood. Here, we performed mass spectrometry-based analyses of cervicovaginal fluids collected from women with differing ovarian hormone levels as demonstrated by menstrual cycle phase or exogenous progesterone-only contraceptive use. We found that proteins associated with maintaining the integrity of epithelial barrier were enriched during times of high estradiol, whereas during times of high progesterone, there was a loss of barrier integrity proteins and an enrichment of proteins with known roles in inflammatory processes including leukocyte infiltration. Progesterone-based proteomic profiles were also strongly associated with neutrophil signatures with some evidence of CD4+ T cell signatures. This study generates new hypotheses about the potential mechanisms of hormone-associated HIV susceptibility including a weakened epithelial barrier and increased HIV target cell recruitment during times of increased progesterone. / February 2016

Orientador: Cezinande de Meira / Banca: João Carlos Pinheiro Ferreira / Banca: Césa Ferraz Jacob / Resumo: O presente estudo teve como objetivo verificar o efeito da aspiração de folículos 25mm sobre a concentração de progesterona plasmática em éguas. O crescimento folicular foi acompanhado diariamente e um único folículo ovariano foi aspirado, exceto quando uma co-dominância foi observada. Neste caso, ambos os folículos foram aspirados no mesmo momento quando atingiram o diâmetro esperado nos respectivos grupos: F ³25mm (n = 6), F ³30mm (n = 6), F ³35mm (n = 6), F pré-ovulatório (n = 6). O Grupo controle (n = 5) não foi submetido à aspiração, acompanhando-se as ovulações espontâneas. A avaliação ultrassonográfica dos ovários foi realizada a cada 24 horas iniciando-se 48 horas antes da aspiração folicular (D0 = dia da aspiração), continuando até a ovulação subsequente. Para a análise estatística, foi utilizada a análise de variância de perfil seguida do método de Tukey com nível de significância de 5% para todas as variáveis, exceto: (1) animais que responderam à aspiração folicular atingindo concentrações de 2ng/mL de progesterona; (2) presença ou ausência de estrutura lútea visualizada pela ultrassonografia, onde foi utilizado o teste do qui-quadrado. A concentração de progesterona foi >2ng/mL, entre os dias 4,0±0,4 e 7,3±0,5 após a aspiração. O intervalo entre a aspiração e a luteólise foi de 16,0±0,5 a 19,0±1,4 dias e entre a aspiração e a ovulação foi de 17,2±2,8 a 23±0,5 dias. A concentração máxima de progesterona alcançada variou entre 6,4±2,6 e 10,9±1,8ng/mL e ocorreu entre os dias 8,7±3,4 e 11,5±1,2. A aspiração folicular em todos os grupos permitiu que as células foliculares se transformassem em estrutura hormonalmente ativa, produtora de progesterona em níveis compatíveis com o diestro. / Abstract: The present study aims to verify the follicles 25mm ablation effect on plasmatic progesterone concentration in mares. Follicular growth was daily accompanied and only one follicle was ablated, excepted when codominance was observed. At this case, both follicles were ablated at the same moment when achived the expected diameter for the following groups: F ³25mm (n = 6), F ³30mm (n = 6), F ³35mm (n = 6), F pre-ovulatory (n = 6). Control group (n = 5) was not submitted to ablation and the spontaneous ovulations were followed. Ultrasound evaluations of the ovaries were done every 24h beginning 48h after follicular ablations (D0 = day of ablation), continuing until subsequent ovulation. For statistical analysis, profile analysis followed by Tuckey method was used with a significance level of 5% for all variables excepted for: (1) animals that responded to follicular ablation achieving progesterone concentrations of 2ng/mL; (2) presence or absence of a luteum structure observed by ultrasound were analysed by Qui-square test. Interval between aspiration and luteolysis varied from 16,0±0,5 to 19,0±1,4 days and between ablation and next ovulation varied from 17,2±2,8 to 23±0,5 days. Maximum concentration of progesterone varied from 6,4±2,6 and 10,9±1,8ng/mL and occurred between days 8,7±3,4 and 11,5±1,2. In all groups, follicular ablation allowed follicular cells to become an active hormonal structure, which produced progesterone in concentrations similar to diestrous. / Mestre

Mammalian ovulation is a complex, hormone-dependent developmental program in which several events must take place in an ordered progression to ensure that the oocyte is competent for fertilization. This process requires the coordinated expression of many genes which must be turned on and off in the right place at the right time for proper development of the follicle. While the hormone signals from the brain that initiate ovulation are known, the master control genes which regulate this process are not well known. Homeobox proteins are potential candidates to perform as master regulators. Homeobox proteins are DNA-binding proteins that regulate the transcription of downstream genes and thereby control biological events. We recently identified a new homeobox gene cluster on the mouse X chromosome that are only expressed in reproductive tissues. These reproductive homeobox (Rhox) homeobox genes are expressed in the ovary, placenta, testis, and epididymis, and thus are good candidates to regulate both male and female reproductive tissue development and physiology. Rhox gene expression fell into three categories: Class I exhibited peak expression prior to ovulation (0-8 hours after hCG), Class II were predominantly expressed during ovulation (8-16 hours after hCG), and Class III peaked after ovulation. The slightly overlapping windows of peak Rhox gene expression suggest that these genes may regulate specific events during the ovulatory cycle. The founding member of the cluster, Rhox5, is highly expressed in granulosa cells of pre-ovulatory follicles. We previously reported that Rhox5-null female mice are viable and fertile, suggesting that RHOX5 is either not essential for ovulation, or that one of the other RHOX factors may compensate functionally in granulosa cells. In order to identify potentially redundant RHOX factors, we examined the expression patterns of all 32 Rhox genes using an eCG primed, hCG induced superovulation model, in wild-type, Rhox5-null, and heterozygous littermate mice. Expression levels of Rhox1, exhibited peak expression prior to being hormonal primed, was reduced in the Rhox5-null animal. However, Rhox8 mRNA and protein were reduced at 2h and 4h post hCG, but recovered once the follicles passed the antral stage of development. Conversely, in progesterone receptor knockout mice (PRKO), Rhox8 exhibited normal stimulation by eCG, but failed to reach its peak mRNA level at 8h post-hCG found in WT mice. This suggests a model in which Rhox8 transcription is dependent upon RHOX5 during early folliculogenesis and progesterone during the periovulatory window when RHOX5 normally wanes. Subsequent promoter analysis in granulosa cells revealed essential homeobox binding and progesterone response elements within Rhox8's 5'-flanking region. Transfection of RHOX5 and PGR expression plasmids stimulated, whereas dominant negative and mutant constructs inhibited, Rhox8 promoter activity. At present, the specific impact of misregulation of Rhox5 and Rhox8 during early folliculogenesis is not known. However, follicle counts from serially sectioned ovaries, extirpated from normal cycling animals, indicated that Rhox5-null mice possess ~50% fewer follicles than heterozygous littermates. Loss of RHOX5 in Sertoli cells results in male subfertility characterized by poor germ cell survival due in part to the misregulation of metabolism promoting genes. One of these genes, Ins2, is also stimulated by RHOX5 and RHOX8 in granulosa cells, suggesting impaired insulin signaling may contribute reduced follicle development in Rhox5-null ovaries.

Traumatic brain injury (TBI) is a major health problem affecting the adult and pediatric population. Scientists and clinicians are working diligently to discover possible therapeutics for the treatment of TBI. Two possible treatments to deal with TBI include sleep and the administration of progesterone. Yet, there are conflicting results from studies regarding the efficacy of either treatment. Sleep appears to reduce neuroinflammation and reduce axonal damage in the brain following TBI. Sleep deprivation, however, may have neuroprotective effects after TBI. Progesterone has also been shown to have neuroprotective effects following TBI. But, there are no sufficient data from animal studies to determine if progesterone is an effective therapeutic. More research studies will have to be conducted to further understand the role of sleep and progesterone in alleviating TBI.

Traumatic Brain Injury (TBI) is a leading cause of death and disability in the United States for which there are no federally approved pharmacological treatments. Preclinical trials with nicotinamide (NAM) and progesterone (Prog) treatment demonstrate beneficial neuroprotection and recovery of function following TBI. The primary goal of this study was to assess both neuroprotection and recovery of function in an animal model of TBI after combination treatment of both NAM and Prog. Animals received a cortical contusion injury over the sensorimotor cortex and were treated with either nicotinamide (75 mg/kg, i.p. NAM loading dose, 12 mg/kg/hr NAM, s.c. over 72 hrs), Prog (10 mg/kg Prog, i.p. over 72 hrs), NAM and Prog(75 mg/kg, i.p. NAM loading dose, followed by continuous infusion of 12 mg/kg/hr NAM, s.c. over 72 hrs; 10 mg/kg Prog, i.p. over 72 hrs) or Vehicle (75 mg/kg, i.p. sterile saline loading dose, followed by continuous infusion 12 mg/kg/hr sterile saline, s.c. over 72 hrs; 10 mg/kg peanut oil, i.p. over 72 hrs), and compared to a craniotomy only (Sham) group. Following this regimen they were assessed in a battery of behavioral (fine and gross motor, sensory, and cognitive) tasks or a histological assessment at 24 hrs post-injury assessing lesion cavity size, degenerating neurons, and reactive astrocytes. Our results replicate the beneficial effects of treatment with either NAM or Prog demonstrating significant improvements in recovery of function, and a reduction in lesion cavitation, degenerating neurons and reactive astrocytes 24 hours post-injury. The combination treatment of NAM and Prog led to a significant improvement in both neuroprotection at 24 hrs post-injury and recovery of function in sensorimotor related tasks when compared to each individual treatment (NAM or Prog). It is suggested here that further preclinical trials using NAM and Prog as a combination treatment should be done to identify any drug interactions, pharmacokinetics, and a new window of opportunity and proper dosing of this combination treatment.

Introduction: The human menstrual cycle is regulated by sex-steroid hormones, including oestrogen (E), progesterone (P4) and androgens which act by ligand binding to their cognate receptors. Perturbation of the complex series of events governing the menstrual cycle may lead to heavy menstrual bleeding (HMB). This is a common debilitating condition and often associated with uterine fibroids. There remains an unmet need for effective, long-term medical treatment so women avoid surgery and preserve their fertility. Selective progesterone receptor modulators (SPRMs, e.g. ulipristal acetate, UPA) are synthetic ligands that bind the progesterone receptor (PR). Many SPRMs have been developed but only mifepristone (for the management of unwanted pregnancy) and UPA are in current clinical use. UPA is licensed for the intermittent treatment of symptomatic fibroids. SPRMs have potential utility for treatment of HMB as administration rapidly induces amenorrhoea but the mechanisms by which this is achieved are unknown. SPRM administration results in unique endometrial morphological changes (progesterone receptor modulator-associated endometrial changes; PAEC). Despite endometrial unopposed estradiol exposure these morphological changes do not appear to be associated with malignancy or pre-malignancy risk. Indeed endometrial cell proliferation appears reduced despite relative progesterone-antagonism. Based upon findings with other SPRMs it was hypothesised that: (i) administration of UPA would have an endometrial specific effect upon the reproductive tract, with regard to alteration in morphology, localisation of sex steroid receptors (SSR) and cell proliferation.; (ii) administration of UPA would impact upon progesterone-regulated (Pregulated) genes in the endometrium. Methods: The data presented within this thesis are derived from biopsies obtained at hysterectomy from the endometrium, fallopian tubes and cervices of women with symptomatic fibroids administered UPA for 8-15 weeks. Samples were obtained for histological assessment, immunohistochemistry and RNA extraction for subsequent quantitative RT-qPCR of sex-steroid receptors (SSR) and proliferation markers. In addition key P-regulated genes within the endometrium were investigated by RT-qPCR and selected protein expression. To further interrogate the anti-proliferative effect, RNA was extracted from “paired” endometrial biopsies from the same woman in the proliferative phase of the menstrual cycle and following subsequent treatment with UPA for at least eight weeks and microarray gene analyses undertaken. Results: Morphological alteration of the endometrium with UPA administration was consistent with previously published data, but with a higher prevalence than previously described. There was a striking alteration in expression and localization of SSRs, particularly PR and androgen receptor (AR), and alteration of many P-regulated genes, consistent with UPA acting with low progesteroneagonism within the endometrium. There was no alteration of SSR expression within the cervix and proliferation was unchanged. Fallopian tube morphology and SSR expression was consistent with proliferative phase but cell proliferation was reduced following UPA administration, consistent with secretory phase levels. Microarray analyses identified multiple transcripts altered relative to proliferative phase, with GREM2 the most significantly down-regulated gene and MUC1 one of the most significantly upregulated genes. Consistent with low levels of mitotic figures and cell proliferation, the most down regulated KEGG pathway was the cell cycle. Multiple elements within this were subsequently validated (RT-qPCR) and included key regulators of all elements of the mitotic cell cycle, many of which were novel to those previously described following administration of another SPRM, mifepristone. In summary the novel data presented in this thesis considerably extend the data available to date concerning the actions of the SPRM, UPA, on the female reproductive tract, and increases knowledge regarding a compound with promising utility for the management of the debilitating complaint of HMB.