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The anteroventral periventricular (AVPV) nucleus of the hypothalamus integrates estradiol (E2) and progesterone (P4) feedback signals from the ovaries to stimulate gonadotropin releasing hormone (GnRH) neurons and trigger an ovulatory surge in luteinizing hormone (LH). E2 maintains the daily cyclic LH surge and P4 quickly amplifies the surge and limits it to one day. P4 amplification of the surge and rapid signaling in the AVPV may occur through its non-classical progestin receptors. Previous in vitro studies using a microarray analysis with N42 mouse embryonic hypothalamic neurons suggest that progesterone membrane component 1 (Pgrmc1) regulates genes linked to the janus kinase (Jak)/signal transducer and activator of transcription (Stat) signaling pathway. I hypothesized that P4 alters Jak/Stats through Pgrmc1 regulation of one or more Jak or Stat molecules and then performed a set of in vitro and in vivo studies to test this. I transfected N42 cells with either scramble or Pgrmc1 siRNA followed by treatment with either ethanol vehicle control or 10 nM P4 and measured Jak1, Jak2, Stat3, Stat5a, Stat5b, and Stat6 mRNA levels via quantitative polymerase chain reaction (QPCR). Jak1 and Jak2 mRNAs increased with P4 treatments, and this upregulation required Pgrmc1. Silencing Pgrmc1 in the cells also produced an increase in Jak1 and Jak2 mRNA, suggesting that Pgrmc1 constitutively suppressed jak1 and jak2 in the absence of P4. None of the Stats were significantly regulated by P4 or Pgrmc1 silencing. To determine how Pgrmc1 regulates Jak/Stat in vivo, I took AVPV microdissections from Pgrmc1 and Pgrmc2 double conditional knockout (DCKO) mice and looked at gene expression of jak/stat. Transcript levels of Jak2, but not Jak1, were severely downregulated in the DCKO animals and Stat mRNAs were not significantly changed. Discrepancies from in vitro and in vivo data prompted me to analyze the role of the class II progestin and adipoQ (Paqr) receptors in Jak/Stat signaling. P4 treatments and siRNA experiments in N42 cells showed that Paqr8, but not Paqr7, was required for P4 upregulation of Jak1 and Jak2 mRNAs. Overall, these findings show that Pgrmc1 regulates Jak1 and Jak2 synthesis in a P4-dependent and -independent manner that requires interaction with Paqr8.

Heat stress (HS), a stress response in homeotherms mainly due to elevated ambient temperature and failure of effective heat dissipation, causes a substantial negative economic impact to livestock industry worldwide. Reduced feed intake, a typical phenomenon observed during HS, was thought to be the primary driver for the milk production loss. However, accumulating evidence indicates that HS influences animal metabolism and endocrine profiles independent of reduced feed intake. Previous studies comparing heat-stressed lactating cows with control group pair-fed (PF) to the intake of HS group but housed in thermoneutral conditions, in order to eliminate the confounding factors result from differentiated feed intakes, showed that HS increased circulating insulin and decreased plasma non-esterified fatty acid (NEFA) in lactating cow, the opposite responses typical of PF cohorts. Therefore, the present studies were performed in order to elucidate the mechanism(s) underlying these counterintuitive changes. In response to a glucose tolerance test (GTT), both HS and PF decreased whole body glucose disposal rate, a sign of insulin resistance. Only PF decreased skeletal muscle insulin sensitivity in terms of reduced protein kinase B (PKB/AKT) phosphorylation, a downstream protein of insulin receptor (IR), while HS group maintained similar intact insulin responsiveness in the liver and skeletal muscle as thermoneutral conditions. There was a global reduction in gene expression of the enzymes related to lipid metabolism in adipose tissue of heat-stressed cows. Similarly, β-adrenergic signaling, a major stimulator of lipid mobilization, was suppressed in terms of NEFA release response during a chronic epinephrine challenge in HS group. After the challenge, phosphorylations of cAMP-response element binding protein (CREB) and hormone sensitive lipase, both located downstream of β-adrenergic receptor, were decreased in HS, but not in thermoneutral conditions, another indicator of impaired adrenergic signaling. In contrast, IR and AKT phosphorylation were increased in HS conditions indicating insulin signaling may be elevated during HS in adipose. Collectively, HS reduces lipid mobilization and appears to favor glucose utilization via alterations of lipid metabolism and hormones signaling pathways. These unique alterations in HS might shed some light on developing counter-HS approaches in the future. / Ph. D.

The purpose of this experiment is to determine if immature female white mice will react to the gonadotropic hormone from the anterior pituitary of several species of fish. If they can be shown to do so, a second purpose is to establish an assay unit for this fish gonadotropin in order that it may be used with predictable results in inducing extra-seasonal spawning for fresh-water conservation practices.

The Anthropocene epoch in which we are currently living, also known as the Holocene, has brought about unprecedented losses in planet Earth’s biodiversity. Numerous extirpations of floral and faunal species have been influenced by human encroachment and more specifically, the exploitation of such species and the respective habitats in which they reside. It is this notion that has propelled many scientists to take up intellectual arms in an effort to protect these invaluable resources. The purpose of this research was to develop technologies to measure and evaluate various variables that influence animal physiology, specifically in amphibians who represent the most threatened class of all animal taxa. Species-specific knowledge including life history and an understanding of evolutionary traits are often needed to effectively guide the management decisions surrounding any given animal population. Specific objectives of this project were to develop non-invasive methods, such as hormone monitoring, machine learning-aided ultrasonography, and near-infrared spectroscopy (NIRS), to assess vital physiological traits, such as biological sex, reproductive status, and chytrid fungus pathogen detection in threatened amphibian species. The novel technologies developed and applied in amphibians here may provide insights for addressing conservation related questions in other animal as well as plant species. Additionally, automation of physiological monitoring techniques through the use of machine learning methods reduces barrier to entry and enables these technologies to be operated by a larger practitioner base. This research also serves to advance methods surrounding chemometric analyses as it pertains to the discipline of wildlife spectroscopy, where large multivariate datasets require data manipulation strategies to produce robust prediction models for the physiological trait of interest for qualitative or quantitative assessment. To that end, a multi-model framework is provided for optimizing predictive outcomes to address questions relating to wildlife management and conservation initiatives.

The purpose of the quality improvement (QI) project was to examine the relationship between amended nursing education concerning diabetes mellitus (DM) Type 2 self-care management incorporating Tune in, Explore, Assist, Communicate, and Honor (TEACH) and Motivational Interviewing (MI) strategies and techniques and the Glycosylated hemoglobin (HgbA1C) of veteran patients with uncontrolled diabetes. The target sample included the 2 licensed practical nurses and 2 registered nurses assigned to 2 primary care teams, and the 10 purposively sampled patients with uncontrolled DM Type 2 from each team. The nurses' competencies were measured through descriptive comparison before and after nursing education implementation using the instrument Patient Education: TEACH for Success Self-Assessment Questionnaire. The nurses' confidence and their perceived importance of the TEACH and MI skills application and skill assessment for promoting health behavior change were tested inferentially with a paired t test before and after nursing education implementation using the instrument Clinician Importance and Confidence Regarding Health Behavior Counseling Questionnaire. The primary care team developed their skills tailored to each patient's needs, considering the guiding principles and premises of the health belief model (HBM). Patients' self-care management knowledge, skills and confidence were improved. The project decreased the elevated HgbA1C of patients measured after the project initiative. The QI project leads to positive social change by decreasing the number of patients with uncontrolled diabetes among the veteran population. The patients and their providers can develop individualized plans of care for diabetes management by educating, redirecting, and evoking behavioral changes in the veteran patients by using a team approach.

<p>This thesis examined the biochemical role of skeletal muscle mitochondria and metabolic consequences of mitochondrial adaptations to exercise in individuals with poor glycemic control. Mitochondrial dysfunction and/or ectopic lipid accumulation has been implicated in the pathogenesis of metabolic-related diseases such as obesity and type 2 diabetes (T2D). However, whether mitochondrial dysfunction is the cause of insulin resistance and T2D or is a consequence of this disorder remains controversial. Alternatively, pro-inflammatory stress signals initiated through altered secretion of adipocytokines and oxidative stress may be a unifying mechanism underlying insulin resistance and T2D. Furthermore, the impact of exercise on muscle adaptation in insulin-resistant states is not well defined. At rest and prior to exercise training, no evidence of mitochondrial dysfunction or disproportionate intramyocellular lipid (IMCL) accretion was detected in obese, insulin-resistant skeletal muscle biopsy samples <em>vs.</em> healthy, lean age-, and fitness-matched men. In response to exercise training (12 weeks, consisting of 32 sessions of 30-60 min @ 50-70% maximal oxygen uptake [VO₂peak]), there was an increase in mitochondrial oxidative phosphorylation (OXPHOS) capacity, mitochondrial content, and IMCL deposition with sub-cellular specificity. Exercise training also reduced both skeletal muscle and systemic oxidative damage, already elevated in the obese. The improved adipocytokine profile associated with obesity after training also coincided with improvements in glycemic regulation. Patients with genetic mitochondrial mutations, resulting in skeletal muscle mitochondrial dysfunction have an increase prevalence of dysglycemia/T2D. However, when evaluated against age- and activity-matched normoglycemic myopathy controls, no differences in mitochondrial electron transport chain protein subunits, mitochondrial or IMCL density, or level of whole-body insulin resistance was detected. In fact, dysglycemic mitochondrial myopathy patients demonstrated <em>higher </em>skeletal muscle OXPHOS capacity and Akt activation, a key step in insulin-stimulated glucose transport activity as compared with normoglycemic mitochondrial myopathy patients. Interestingly, a significant impairment in β-cell function (defective insulin secretion), in the dysglycemic patients was observed coincident with elevated glucose levels during the oral glucose tolerance test (OGTT). These findings indicate that insulin resistance does not cause skeletal muscle mitochondrial dysfunction/IMCL accumulation or <em>vice versa</em> and provides evidence against a direct link between mitochondrial dysfunction and the development of insulin resistance/T2D. Perhaps, oxidative stress/inflammation and pancreatic β-cell erosion mediate the observed obesity-induced insulin resistance and mitochondrial myopathy-associated T2D, respectively? Twelve weeks of moderate endurance exercise is an effective strategy to improve mitochondrial capacity, oxidative damage, inflammation, and glycemic regulation in insulin-resistant, obese individuals, but an improvement in muscle insulin sensitivity did not appear to be required.</p> / Doctor of Philosophy (Medical Science)

Vitellogenin (Vtg) is a precursor to yolk-proteins produced in the liver of many invertebrates and non-mammalian vertebrates; its synthesis is stimulated by the hormone estradiol (E2). This study is the first to characterize vitellogenin synthesis in a placental viviparous elasmobranch, the yolk-sac placental bonnethead shark, Sphyrna tiburo. This study focused on determining where and when Vtg is produced, as well as what hormonal factors regulate Vtg production. The liver was confirmed as the site of Vtg production via immunohistochemistry. Immunoreactivity was also observed within granulosa cells of ovarian follicles; further experimentation is needed to determine if this is indicative of Vtg production by these cells. Using immunoblotting, the highest proportions of Vtg positive females were found in March, with Vtg production continuing into April and May. Putative Vtg production was found to begin in August for some individuals, with production continuing throughout the fall and winter months. In regards to hormonal regulation, immunohistochemical analysis identified receptors for E2 and progesterone (P4) within the liver. Comparison of the monthly E2 and Vtg cycles provides evidence that E2 stimulates Vtg production in S. tiburo, as high or increasing concentrations of E2 correlated with Vtg presence in the plasma. Preliminary results also suggest in vitro production of Vtg by liver tissue when exposed to E2. Comparison of the monthly P4 and Vtg cycles suggests P4 may inhibit Vtg synthesis, with higher levels of P4 found in the months when Vtg production declines. Additionally, the methods developed for this study were able to identify Vtg in the plasma of other elasmobranch species. Vtg detection in plasma may thus be an ideal new, nonlethal method for characterizing elasmobranch reproductive periodicity, which will aid in assessing population growth and allow for managers to possess more accurate information to make appropriate decisions for the populations.

Shark fisheries are a multimillion dollar industry in the United States and have significant contributions both recreationally and commercially. In order to maintain this industry, fisheries must be properly managed. An understanding of life history and reproduction is crucial in order to adequately manage shark fisheries. The finetooth shark, Carcharhinus isodon, is a member of the small coastal shark (SCS) fishery complex. It is found in Atlantic waters from South Carolina to Florida and throughout the Gulf of Mexico. This species has recently come under increased fishing pressure and has previously been overfished. New data is needed in order to properly assess the reproductive capacity of this species so that healthy populations can be sustained. This study evaluated the overall reproductive cycle of northwest Atlantic populations of C. isodon using composite variables of morphology, histology and endocrinology. Atlantic C. isodon were found to display seasonal reproduction with biennial reproductive periodicity with a 12 month gestation period and litters of 2-6 pups. E2 and T displayed similar patterns and were highest during gametogenesis. P4 did not show any discernible patterns. Estrogen receptors were found in developing follicles of gravid and non-gravid females, in the oviducal glands of non-gravid and early pregnant females and in the uterus of pre-ovulatory females. P4 was found in the oviducal glands in all reproductive stages and in the uterus of pre-ovulatory females. Androgen receptors were found in Sertoli cells and mature spermatozoa as well as epididymal epithelial cells.

Metabolic hormones and their axes, including the target tissues and receptors, regulate the tissue specific utilization of nutrients with in the body. The purpose of this research was to understand the hormonal control of complex nutrient partitioning mechanisms involved in young, growing animals. Specifically, this involved the investigation of metabolic hormones and the regulation of growth in two common species of phocids (true seals): harbor seal (Phoca vitulina) and Northern elephant seal (Mirounga angustirostris) pups. This longitudinal study examines young phocids from nutritional nadir through realimentation (realimentation) to investigate how metabolic hormones involved in both food intake and nutrient partitioning change with respect to nutritional state. To investigate the role of metabolic hormones during realimentation in a small phocid seal, chapter 2 focuses on the changes in the somatotropic axis and ghrelin during a 10-week period of realimentation following nutritional nadir. Chapter 3 focuses on the application of the results of previous research and the second chapter of this thesis to a specific experimental feeding project. Chapter 4 focuses on the response to changes in nutritional status in the fasting adapted NES. Given the changes in metabolism and priority of nutrient utilization associated with transitioning from a nursing neonate to a fasting adapted juvenile, NES provide a unique opportunity to assess the effect of age on the response to realimentation. Overall, this research will further expand the understanding of tissue specific demands and the effect on endocrine response to realimentation. By incorporating assessments of metabolic changes based on nutrition as well as age, this study will expound on how metabolic hormones are involved in regulating the trade-off between adipose and lean tissue development in this unique taxon.