Research indicates a relationship between prenatal maternal stress (PNMS) and attention levels in children. Studies with non-human primates suggest that both exposure to PNMS and the timing of a stressor during pregnancy are associated with greater attention problems in offspring; however, this has not been sufficiently tested in humans because of methodological constraints. We explored the relationship between objective and subjective measures of PNMS for 85 women who were pregnant during the 1998 Quebec Ice Storm and attention levels in their 5.5 year-old children, while controlling for socioeconomic status, child's sex, and maternal state anxiety. As a group, children in the study displayed significantly less attentive behaviour compared to a standardized norm. Boys in this group were significantly less attentive than girls. Objective PNMS exposure during 1st trimester correlated positively with attention problems in the children, accounting for 15.2% of the variance in attention levels as rated by Kindergarten teachers. In a behavioural task to detect sustained attention difficulties and Attention Deficit-Hyperactivity Disorder (ADHD) symptoms, children exposed during the 3rd trimester obtained significantly worse scores compared to children exposed in other trimesters. However, in this case, objective and subjective PNMS appeared to act as protective factors, raising the possibility of the presence of another unknown mechanism that negatively affected the attention variables.

Pregestational and gestational diabetes (GDM) places the mother and her offspring at an increased risk for later development of insulin resistance and type 2 diabetes. Oxidative stress may mediate long-term disturbances in glucose homeostasis associated with type 2 diabetes and the metabolic syndrome. This thesis describes a cross-sectional study examining serum concentrations of free radical generated oxysterols as markers of oxidative stress in a cohort of teenage daughters from pregnancies with and without GDM. Daughters of GDM-pregnancies had a tendency of higher levels of serum oxysterols (7beta-hydroxycholesterol); however, this difference was not statistically significant after adjustment for total cholesterol. Serum oxysterols were significantly correlated with obesity measures such as waist circumference and BMI, which likely accounted for the tendency for higher measures of oxysterol concentrations in the GDM daughters. Oxysterols represent potentially important biomarkers for oxidative stress in adolescent girls as their levels track with the metabolic syndrome risk factors. / Le diabète pré-gestationnel et le diabète de gestation (DG) augmentent le risque dedéveloppement d'une future résistance à l'insuline et de diabète de type 2 autant pourla mère que pour l'enfant. Le stress oxydatif est un facteur potentiel impliqué dans ledéséquilibre du glucose sanguin associé au diabète de type 2 et au syndromemétabolique. La présente thèse est une étude sectionnelle croisée, ayant pour but demesurer des marqueurs du stress oxidatif, notamment la concentration des oxystérolsgénérés par les radicaux libres dans le sérum d'adolescentes, nées de mères ayantprésenté ou non un diabète de gestation. Nos résultats montrent des concentrationsd'oxystérols (7P-hydroxycholesterol) plus élevées dans le sérum de filles issues degestations diabétiques à comparer aux filles de mères n'ayant pas eu de DG.Cependant, la différence entre les deux groupes n'était pas statistiquementsignificative après un ajustement au cholestérol total. La concentration d'oxystérolsétait significativement corrélée aux marqueurs d'obésité, notamment la circonférencede la taille et l'index de masse corporelle, possiblement à l'origine de la tendance desoxystérols à être plus élevés dans le cas des adolescentes issues de gestationsdiabétiques.

Biomarkers measured in the blood can provide information about disease pathophysiology, diagnosis and prognosis. Pronounced proinflammatory responses are characteristic of severe malaria, and excessive activation of the immune system is central to the pathophysiology of both cerebral malaria and placental malaria. Severe malaria is characterized by cytoadherence of parasitized erythrocytes to the microvasculature; impaired tissue perfusion; dysregulated inflammatory responses; and activation of the complement system, mononuclear cells, and endothelium. Despite the availability of effective antimalarial drugs, the mortality rate in severe malaria remains unacceptably high. To glean further insight into malaria pathophysiology, we investigated host biomarkers of immune activation in the blood of subjects with different manifestations of severe disease. C5 has been identified as being necessary and sufficient for the development of experimental cerebral malaria. We hypothesized that C5a (a terminal component of the complement cascade with potent inflammatory properties) may mediate its action by inducing and exacerbating inflammatory processes in severe malaria, leading to endothelial activation and dysregulated angiogenesis. I tested this hypothesis in vitro, and found that C5a interacted with malaria toxin PfGPI to drive deleterious inflammatory and anti-angiogenic responses. As C5a and anti-angiogenic factor sFlt-1 have been implicated in models of pathologic pregnancies, we asked whether increased levels of C5a in subjects with placental malaria were associated with altered angiogenesis and poor birth outcomes. Our results suggest that C5a impairs angiogenic remodelling in placental malaria leading to vascular insufficiency and fetal growth restriction. Further, altered profiles of inflammatory and angiogenic biomarkers in the periphery may identify occult placental malaria infections. We extended these observations to cerebral malaria where similar pathogenic pathways contribute to disease pathophysiology. In adults and children with cerebral malaria, altered profiles of angiogenic proteins were associated with disease severity and mortality and represent putative diagnostic and prognostic biomarkers in severe malaria.

The incidence of preterm labor has risen over recent decades and preventative antibiotic treatment is ineffective. Associated with a 40% increased risk of preterm birth, bacterial vaginosis is characterized by a decrease in lactobacilli and increase in pathogenic bacteria in the vaginal flora. Ascent of bacterial products to the intrauterine environment stimulates cytokine and prostaglandin secretion from invading immune cells and gestational tissues. Probiotic lactobacilli modulate the immune responses in mouse macrophages and human placental trophoblast cells. The focus of this thesis was to determine the influence of Lactobacillus rhamnosus GR-1 (GR-1) on cytokines and prostaglandins which are part of the activated pathway in infection and/or inflammation mediated preterm labour. GR-1 increased amnion chemokine and reduced pro-inflammatory cytokine release. GR-1 elevated prostaglandin E2 release that was paralleled by an increase in mPGES2 expression. It is possible that t that GR-1 may enhance the host defense barriers of the amnion to pathogenic bacteria.

Biomarkers measured in the blood can provide information about disease pathophysiology, diagnosis and prognosis. Pronounced proinflammatory responses are characteristic of severe malaria, and excessive activation of the immune system is central to the pathophysiology of both cerebral malaria and placental malaria. Severe malaria is characterized by cytoadherence of parasitized erythrocytes to the microvasculature; impaired tissue perfusion; dysregulated inflammatory responses; and activation of the complement system, mononuclear cells, and endothelium. Despite the availability of effective antimalarial drugs, the mortality rate in severe malaria remains unacceptably high. To glean further insight into malaria pathophysiology, we investigated host biomarkers of immune activation in the blood of subjects with different manifestations of severe disease. C5 has been identified as being necessary and sufficient for the development of experimental cerebral malaria. We hypothesized that C5a (a terminal component of the complement cascade with potent inflammatory properties) may mediate its action by inducing and exacerbating inflammatory processes in severe malaria, leading to endothelial activation and dysregulated angiogenesis. I tested this hypothesis in vitro, and found that C5a interacted with malaria toxin PfGPI to drive deleterious inflammatory and anti-angiogenic responses. As C5a and anti-angiogenic factor sFlt-1 have been implicated in models of pathologic pregnancies, we asked whether increased levels of C5a in subjects with placental malaria were associated with altered angiogenesis and poor birth outcomes. Our results suggest that C5a impairs angiogenic remodelling in placental malaria leading to vascular insufficiency and fetal growth restriction. Further, altered profiles of inflammatory and angiogenic biomarkers in the periphery may identify occult placental malaria infections. We extended these observations to cerebral malaria where similar pathogenic pathways contribute to disease pathophysiology. In adults and children with cerebral malaria, altered profiles of angiogenic proteins were associated with disease severity and mortality and represent putative diagnostic and prognostic biomarkers in severe malaria.

The incidence of preterm labor has risen over recent decades and preventative antibiotic treatment is ineffective. Associated with a 40% increased risk of preterm birth, bacterial vaginosis is characterized by a decrease in lactobacilli and increase in pathogenic bacteria in the vaginal flora. Ascent of bacterial products to the intrauterine environment stimulates cytokine and prostaglandin secretion from invading immune cells and gestational tissues. Probiotic lactobacilli modulate the immune responses in mouse macrophages and human placental trophoblast cells. The focus of this thesis was to determine the influence of Lactobacillus rhamnosus GR-1 (GR-1) on cytokines and prostaglandins which are part of the activated pathway in infection and/or inflammation mediated preterm labour. GR-1 increased amnion chemokine and reduced pro-inflammatory cytokine release. GR-1 elevated prostaglandin E2 release that was paralleled by an increase in mPGES2 expression. It is possible that t that GR-1 may enhance the host defense barriers of the amnion to pathogenic bacteria.

The seasonal termination of ovarian cycles in mares initiated near the time of the autumnal equinox is a significant managerial issue for horse breeders world-wide. Studies presented herein had two over-arching aims. In Aim I, objectives were to develop the principals needed to apply gonadotropin-releasing hormone (GnRH) therapeutics for routinely establishing pregnancies in the winter anovulatory mare. We first tested the hypothesis that continuous administration of native GnRH, beginning in either early February or March, would induce ovulation without reversion to an anovulatory state following treatment withdrawal. Continuous 28-d treatments elevated circulating luteinizing hormone (LH) and stimulated spontaneous ovulation much earlier than controls. However, mares treated only in February ceased ovarian cycles at termination of treatment. In contrast, mares administered GnRH in March continued to exhibit estrous cycles. Thus, we concluded that GnRH treatment must continue through March to ensure continued escape from winter anovulation. We then tested the hypothesis that the Julian day of conception could be accelerated in winter anovulatory mares treated continuously with native GnRH for 56 d beginning on February 1. Indeed, GnRH treatment caused a marked increase in the frequency of pregnancy compared to controls. Data illustrated that continuous administration of native GnRH is a practical and highly efficient option for managing seasonal anovulation. In Aim II, we examined hypothalamic distribution, adenohyphyseal receptor gene expression, and ligand functionality of RFRP3 in the mare during the breeding and non-breeding seasons. Hypothalamic RFRP3 mRNA was detected in the mare; however, neither hypothalamic expression of RFRP3 nor its anterior pituitary receptor differed between reproductive states. We then used equine adenohypophyseal cell culture to test the hypothesis that RFRP3 reduces the responsiveness of the equine gonadotrope to GnRH. Addition of RFRP3 to cell culture failed to counter the effects of GnRH. Finally, the effects of a RFRP3 receptor-signaling antagonist (RF9) were examined in winter anovulatory mares. A robust increase in circulating concentrations of LH relative to controls was observed in response to RF9 treatments, but treatments had no effect on adenohypophyseal responsiveness to GnRH. Data provide indirect evidence that antagonism of the RFRP3 system by RF9 may be at the GnRH neuronal level.

Human Cytomegalovirus (HCMV) is the most common cause of congenital infection in newborns. One mechanism for this virus to reach the fetus is to cross the placenta through the syncytiotrophoblast layer. Accumulation and protection of pathogens in the syncytiotrophoblast could affect the systemic distribution of pathogens and prolong maternal infections leading to increased incidence of fetal infections. Primary infections, reactivation or reinfection with another strain during pregnancy are risk factors for intrauterine HCMV transmission to the fetus. All lead to an active infection; however, viral load in blood or urine does not correlate with intrauterine transmission. I have shown that HCMV reversibly binds to the syncytiotrophoblast in vitro, protecting it from degradation. Furthermore, I demonstrated in vivo that HCMV is present in the placenta, even when cleared from maternal blood and urine. This evidence suggests increased potential for fetal transmission by virtue of continued virus localized at the maternal-fetal interface.

Herbal and homeopathic compounds have been used to aid in childbearing and pregnancy for centuries. Much of this information is anecdotal and lacks scientific support, making it difficult to evaluate safety and efficacy. Increased public interest in alternative treatments leads to the need for a systematic review on the topic. Herbal remedies are most often used to treat the most common pregnancy-related problems like nausea, stretch marks and varicose veins. In contrast to this, concerns have also been raised about the adverse effects of these remedies. The primary objective of this research project was to conduct a systematic review to assess the maternal and neonatal outcomes of ingested herbal and homeopathic remedies using during pregnancy, birth and breastfeeding.

Ethanol consumption during pregnancy can result in wide range of negative outcomes, including pre-and post-natal mortality, growth retardation, physical abnormalities and brain deficits, manifested as behavioural impairments. These outcomes can result from “binge-drinking” (generally defined as >5 standard drinks on a single occasion) or chronic ethanol consumption. Ethanol-induced zinc (Zn) deficiency is one of the mechanisms proposed as a cause of ethanol teratogenicity. We have previously demonstrated in mice that ethanol exposure on gestational day (GD)8 (during organogenesis) can alter Zn homeostasis by inducing the Zn-binding protein metallothionein (MT) in the maternal liver. This causes plasma Zn concentrations to decrease as Zn redistributes into the liver, and consequently decreases the fetal Zn supply and increases the risk of teratogenicity. Subcutaneous Zn treatment with ethanol on GD8 can prevent the deleterious effects of ethanol on the fetus (i.e. physical abnormalities and spatial memory impairments). The main objective of this thesis was to investigate whether a less invasive approach of giving dietary Zn supplementation throughout pregnancy could provide similar protective benefits against a range of adverse outcomes caused by prenatal binge or chronic ethanol exposure. Binge ethanol exposure in early pregnancy (i.e. where mice are injected with 25% ethanol (0.015 ml/g) intraperitoneally at 0 and 4 hours on GD8) significantly increased the incidence of birth abnormalities measured on GD18. These included craniofacial abnormalities (microphthalmia, anophthalmia) and limb defects. Ethanol also increased postnatal mortality between birth and postnatal day (PD)60. In a separate study, offspring from dams given ethanol on GD8 were subjected to a physical and behavioural screening protocol (including tests for vision, olfactory, exploratory, anxiety and motor impairments) and subsequently a cohort of phenotypically-normal offspring were randomly selected for testing in a cross-maze escape task (for spatial learning and memory) and an object recognition test (for short-term non-spatial memory). While ethanol did not affect behaviour measured during screening, it resulted in spatial memory and object recognition memory impairments in adult offspring. The most important finding was that dietary Zn supplementation throughout pregnancy significantly increased plasma Zn concentrations at the time of ethanol exposure (avoiding the “typical” ethanol-induced decrease in plasma Zn) and prevented all negative outcomes resulting from early ethanol exposure (birth abnormalities, mortality, spatial and object recognition memory impairments). In the chronic ethanol mouse model (i.e. where mice were fed a liquid diet containing 27 % v/v ethanol-derived calories from GD6-18), ethanol did not affect offspring growth between birth and PD21 or spatial memory in adult offspring, thus, the influence of Zn supplementation could not be examined for these parameters. While ethanol decreased offspring weight at PD50 and increased mortality between birth and PD40, they were not prevented by Zn supplementation throughout pregnancy. The findings from this thesis emphasise that organogenesis is a particularly vulnerable period to ethanol exposure and even a binge of ethanol during this time can result in dysmorphology, mortality and spatial and object memory impairments in adulthood. In addition, dietary Zn supplementation is protective against the deleterious effects of binge ethanol exposure in early pregnancy. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1368113 / Thesis (Ph.D.) - University of Adelaide, School of Molecular and Biomedical Sciences, 2009