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1

Sivakumar, Kavitha. "The impact of maternal obesity and gestational diabetes mellitus on adipose tissue and placental derived adipocytokines." Thesis, University of Warwick, 2013. http://wrap.warwick.ac.uk/58490/.

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Pregnancy; a natural insulin resistant state; becomes exaggerated when complicated by obesity and gestational diabetes (GDM). Both obesity and GDM are associated with severe maternal and fetal complications as well as with increased risks of obesity in the offspring later in life. Little work has been performed on the levels of adipokines in lean, obese and diabetic pregnancy. This study aimed to explore the roles of three adipokines; namely, Adipsin, Acylation stimulating protein and Fibroblast Growth Factor-21, all of which are involved in insulin resistant and dysmetabolic states such as obesity and type 2 DM. We hypothesized that these adipokines might play a role in pregnancy. A cohort of Caucasian pregnant women undergoing elective caesarean section was studied. Clinical parameters were assayed as well as circulating maternal and fetal levels of adipsin, ASP and FGF21. Paired samples of fat and placental tissue were taken for explant studies to measure secreted Adipsin, ASP and FGF21 levels. Cord levels of adipsin and ASP were significantly elevated in the offspring of obese and diabetic mothers compared to their lean controls. Plasma FGF21 levels were significantly higher in GDM compared to lean controls. FGF21 levels in cerebrospinal fluid (CSF) were also measured and a CSF/Plasma ratio calculated. I have identified the human placenta as a source of adipsin, ASP and FGF21. More specifically, I have shown that placental Hofbauer cells (macrophages) produce adipsin and ASP. This is the first time secretion of adipsin and ASP by Hofbauer cells has been demonstrated. I conjecture a role of these macrophages in lipid metabolism at the materno-fetal interface. Also, I describe that GDM mothers have higher CSF FGF21 as compared to controls but the CSF:plasma ratio of FGF21 was lower in GDM mothers, potentially suggesting an alternative reason for and contributing to hyperglycaemia in GDM.
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2

Bawazeer, Nahla M. "Vitamin B12 and folate status during pregnancy among Saudi population." Thesis, University of Warwick, 2011. http://wrap.warwick.ac.uk/49940/.

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T2DM is a growing health problem worldwide. It is now increasingly being diagnosed earlier in life. The factors involved in such an epidemic are complex. The intrauterine environment has long been known as an important contributor to many diseases including metabolic disorders such as T2DM. Recently, there is emerging evidence for maternal micronutrients affecting vital developmental processes in utero which can adversely “programme” the offspring to develop metabolic disorders in later life. Thus, “gene-diet” interaction during foetal development is likely to be a significant contributor to the epidemic of T2DM. In particular, the intrauterine imbalance between the two related vitamins, vitamin B12 and folate, affect DNA methylation and in turn programme the foetus for the whole life. Evidence from mandatory folic acid fortification studies suggests that in the presence of adequate folate, neural tube defects due to vitamin B12 insufficiency have tripled. In India, children born to mothers with “high folate and low vitamin B12” had higher adiposity and insulin resistance. Therefore, micronutrient status during pregnancy is likely to have a significant impact on the metabolic risk of the offspring. This thesis examines whether vitamin B12 insufficiency is prevalent in pregnancy, especially in a non-vegetarian population across the world as well as the Saudi pregnant population. As estimated intake is an accepted measure for micronutrient levels, we also examined the relationship between estimated vitamin B12 and folate intake with actual levels in the blood. We have found that vitamin B12 insufficiency was not uncommon during pregnancy across the world even in the non-vegetarian population and is also common in the Saudi population. Surprisingly, vitamin B12 insufficiency was observed in 50% of the tested population even in the presence of adequate vitamin B12 intake. In addition, we have also shown for the first time in the Saudi population that maternal BMI is inversely related to vitamin B12 levels, particularly in pregnancy. Even though we have shown a similar (or worse) picture in mothers with gestational diabetes, this study needs to be replicated, as our numbers are too small. Prospective studies linking the role of vitamin B12 insufficiency especially in the presence of high folate on birth outcomes in the Saudi population as well as intervention studies investigating the role of vitamin B12 supplementation in women of childbearing age and in pregnancy are urgently needed.
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3

Chen, Li. "The potential use of cAMP and progesterone in the prevention of preterm labour." Thesis, University of Warwick, 2012. http://wrap.warwick.ac.uk/50049/.

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There is increasing evidence that cAMP promotes the relaxation of the myometrium, and other types of smooth muscle via the activation of cAMP-dependent protein kinase and other intracellular signalling pathways. The data from the first part of this study show that cAMP reduces the expression of most of the labour-associated gene, such as oxytocin receptor (OTR) (via PKA) and FP receptor. Interestingly, cAMP induces COX-2 expression via MAPK pathway. Therefore, with the exception of the COX-2 data, these findings suggest that cAMP agonists have the potential to be effective tocolytic agents. Most researchers also believe that progesterone plays an important role in maintaining uterine quiescence throughout pregnancy by inhibiting the expression of contraction-associated proteins in the myometrium. However, the exact mechanism responsible for the progesterone-induced inhibition of labour-associated genes expression remains unclear. Clinically, progesterone treatment has been shown to reduce the risk of preterm labour (PTL) in high-risk populations. Surprisingly, progesterone had no effect on the occurrence of preterm labor in multiple pregnancies. So the second part of this project focused on the hypothesis that cAMP could enhance the myometrial response to progesterone. The combined effects of cAMP and progesterone on the activation of different pathways in primary cultures of human myometrial cells was studied. It was found that forskolin enhanced the progesterone-induced expression of genes such as FKBP5 and 11βHSD1. The forskolin effects included the up-regulation of progesterone receptor (PR)-B levels and enhanced progesterone-driven activity of a progesterone response element (PRE) as well as increased PR-B binding to the PRE. The data also show that forskolin attenuate the association between PR and NCoR. Knockdown of NCoR blocks the ability of forskolin to enhance progesterone driven PRE activity. Furthermore, the ability of progesterone to repress IL-1β-induced COX-2 expression was enhanced by forskolin, which was associated with a delay in IL-1β-induced nuclear phospho-p65 entry, as well as an increase in IκB synthesis and a reduction in NF-κB binding to the COX-2 promoter. Overall, these data identify important interactions between cAMP and progesterone pathways that control myometrial expression of labour-associated genes and point towards a novel role for cAMP agonists to supplement progesterone-based clinical protocols in the treatment of PTL.
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4

Vasilopoulou, Elisavet. "The role of thyroid hormones in placental development and the importance of the thyroid hormone transporter MCT8." Thesis, University of Birmingham, 2010. http://etheses.bham.ac.uk//id/eprint/1146/.

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Thyroid hormones (THs) are important for fetoplacental development. Human intrauterine growth restriction (IUGR) is associated with malplacentation and reduced fetal circulating concentration of THs. The expression of the plasma membrane TH transporters MCT8, MCT10, LAT1, LAT2, OATP1A2 and OATP4A1 was characterised in human placental biopsies across gestation. The protein expression of MCT8 was increased in samples from severe IUGR compared with normal pregnancies. In vitro, triiodothyronine (T\(_3\)) decreased survival and increased apoptosis of IUGR compared with normal cytotrophoblasts, which was associated with increased MCT8 expression. In normal cytotrophoblasts, MCT8 upregulation decreased survival, whilst MCT8 silencing increased survival independently of T\(_3\). In the extravillous trophoblast-like cell line, SGHPL-4, T\(_3\) resulted in a significant increase in cell invasion when MCT8 was upregulated. Contrary to cytotrophoblasts, silencing MCT8 decreased apoptosis in SGHPL-4s independently of T\(_3\). In mice, fetal to placental weight ratio was decreased in male MCT8-null compared with wild-type embryos. These findings support the hypothesis that THs have an important role in fetoplacental development and that IUGR is associated with changes in TH transport and responsiveness of the placenta. Furthermore, they highlight the importance of MCT8, which impacts on placental cells via both T\(_3\)- dependent and independent mechanisms.
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5

Day, Priscilla Elly Liwawa. "The transfer and metabolism of glucose and amino acids by the human placenta." Thesis, University of Southampton, 2012. https://eprints.soton.ac.uk/377713/.

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6

Salter, Scarlett. "Embryo signals for successful implantation." Thesis, University of Warwick, 2016. http://wrap.warwick.ac.uk/80221/.

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Human pre-implantation embryos display a high prevalence of aneuploidy and chromosomal mosaicism, unique from any other species. The decreasing incidence of aneuploidy observed between the cleavage and blastocyst stages of preimplantation embryo development infers a degree of 'self-correction' following activation of the embryonic genome. However, contrary to the previous assumption that only euploid embryos should be considered 'normal', new evidence has confirmed that mosaic embryos can result in the birth of healthy babies. Thus, aneuploidy should be viewed as an intrinsic feature of human pre-implantation embryo development, which presents a novel challenge at implantation. The endometrium must implement both positive and negative selection, in order to limit maternal investment to only viable embryos. The ability of the endometrium to act as a 'biosensor' of embryo quality has been well documented yet there is little direct evidence for the key regulators of this process. For the first time, we demonstrate a biological context for embryo biosensoring. Firstly, we discover novel embryo-secreted proteases that are enhanced at the blastocyst stage and relate to implantation outcome upon embryo transfer. Secondly, we identify corresponding protease-sensitive receptors in the endometrium, heightened during the window of implantation. By demonstrating the cleavage and de-activation of endometrial toll-like receptor 4 (TLR4) by embryo conditioned medium, we link successful implantation to a diminished inflammatory response. Furthermore, we demonstrate that TLR4 levels in the endometrium constitute a selectivity checkpoint, which is supressed in women suffering from recurrent miscarriage (RM).
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7

Bhandari, Harish. "Obesity and peri-implantation endometrium." Thesis, University of Warwick, 2014. http://wrap.warwick.ac.uk/65254/.

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Obesity is a global health problem and the current available evidence from the literature suggests that obese women may suffer from a wide spectrum of reproductive complications. The current understanding of obesogenic effects on the peri-implantation endometrium is limited and has become an important research topic as the emerging clinical evidence from the published studies indicate the possible role of the endometrium. The first part of this thesis addresses the clinical question of whether an early pregnancy outcome is affected by the body mass index and whether there is any difference in time taken to achieve pregnancy in obese women with recurrent miscarriage when compared to normal weight women. The results are in chapters 3 and 4, where we demonstrated that obese women were more likely to have miscarriage of empty gestational sac or anembryonic pregnancies. In recurrent miscarriage context, the obese women were more ‘super-fertile’ suggesting the possible loss of an endometrial ability to select normal from abnormal pregnancies. The second part of this thesis provides an analysis of the peri-implantation endometrial stromal compartment in normal weight and obese women. In chapter 5, using immunohistochemical methods it was shown that there was no difference in the uterine natural killer cell and macrophage density in the peri-implantation endometrium of different weight groups. This suggests that the endometrial dysfunction in obese women with reproductive failure does not appear to be immune cell mediated. In chapter 6 it was shown that the clonogenecity of endometrial mesenchymal stem cells (W5C5+) was significantly negatively correlated with the BMI. The obese women had significantly reduced cloning efficiency of W5C5+ cells when compared to normal weight women, suggesting of a possible sub-optimal regenerative capacity of the endometrium in obese women. Finally, chapter 7 showed a potential association between obesogenic environment and impaired stromal cell decidualisation. Using an in-vitro model, it was shown that there was no significant difference in the expression of decidualisation markers (PRL and IGFBP1) in the decidualising endometrial stromal cells from normal weight women when compared to high BMI women. However, when the stromal cells were decidualised in an artificial obesogenic environment, the PRL expression was significantly inhibited in the presence of supernatant from adipose tissue explants of obese women when compared to normal weight women. In summary, the findings from my work have provided an understanding of the peri-implantation endometrium in obese women and evidence to suggest that the endometrial stromal function is possibly facilitated by metabolic influences.
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8

Cavilla, Jennifer Louise. "The effects of factors influencing human oocyte maturation upon fertilization and preimplantation embryo development." Thesis, University of Warwick, 2002. http://wrap.warwick.ac.uk/73509/.

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The competence of oocytes to mature and undergo fertilization and embryonic development is known to be influenced by the conditions under which their maturation occurs. Suboptimal maturation in vitro currently limits the use of immature oocytes for embryo creation. This project examines the relationship between the conditions of in vitro maturation of human oocytes and aspects of their subsequent developmental competence through the in vitro creation and analysis of research embryos. This work is essential in defining effective and safe conditions for the use of human immature oocytes in programmes of clinical treatment to alleviate infertility. Human immature oocytes were exposed in vitro to various concentrations of meiosis activating sterol (FF-MAS), an endogenous mediator of follicle and oocyte function, or epidermal growth factor (EOP), in the absence of other hormonal support. Their survival and further development relative to controls were measured by assessing the proportions maturing, fertilizing by sperm injection (ICSI), and cleaving in vitro. Image analysis was used to measure various dimensions of oocytes and embryos daily. A pilot study of chromosome and spindle configurations at meiotic metaphase was also conducted. The major findings of this project are that FF-MAS supplementation of maturation medium has different positive effects upon immature oocytes arising from patient groups having different endocrine profiles and yielding differing oocyte populations. FF-MAS at 30J.lg/ml promotes survival of oocytes from unstimulated patients with polycystic ovaries (p
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9

Venkatakrishnan, R. "Role of decidual corticosteroid production in reproductive failure." Thesis, University of Warwick, 2014. http://wrap.warwick.ac.uk/62785/.

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Glucocorticoids have been implicated in many processes including successful embryo implantation, placentation, and the growth and development of the fetus. Glucocorticoid treatment has been advocated as a treatment to improve reproductive outcome for a number of reasons. Prednisolone treatment has been associated with improvement in clinical outcome in women with recurrent miscarriage and improvement in outcome of In Vitro Fertilisation. Steroids have been found to reduce the high levels of uterine natural killer cells which have been associated with recurrent miscarriage and recurrent implantation failure. Glucocorticoids stimulate peri-implantation human chorionic gonadotrophin secretion from trophoblast of early human embryo and accelerate trophoblast growth and invasion. Elevated uterine NK cell levels during the implantation window are associated with reproductive failure and can be repressed by oral glucocorticoids. We have shown that decidualizing human endometrial stromal cells profoundly up regulate the expression and activity of 11beta-hydroxysteroid dehydrogenase type 1, the enzyme that converts inert cortisone to active cortisol; thus establishing a local cortisol gradient and activation of glucocorticoid and mineralocorticoid receptors. We also found that elevated levels of uterine natural killer cells in the stroma underlying the surface epithelium of endometrium are associated with defective decidualization of resident stromal cells, inadequate cortisol biosynthesis and suboptimal induction of corticosteroid-dependent enzymes involved in lipid droplet accumulation and retinoid transport pathway. Thus, impaired decidualization limits the induction of a local cortisol gradient in the stroma underlying the surface epithelium. This in turn accounts for possible inappropriate recruitment of uterine natural killer cells and suboptimal expression of metabolic genes involved in lipid biosynthesis and retinoid storage pathway. Based on the findings, we postulate that patients suffering recurrent miscarriage associated with high uterine NK cell density may benefit from corticosteroid treatment in early pregnancy; although this assumption will need to be tested in a larger clinical trial.
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10

Morgan, Hannah Louise. "Pregnancy in the stroke-prone spontaneously hypertensive rat : investigating impaired vascular remodelling and establishing a novel model of superimposed pre-eclampsia." Thesis, University of Glasgow, 2018. http://theses.gla.ac.uk/30989/.

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Hypertensive disorders of pregnancy are an increasingly common disorder in modern society. The increasing prevalence of women with pre-eclampsia superimposed on a background of chronic hypertension is a significant burden in modern society and is profoundly detrimental to both mother and child; during pregnancy and beyond. Little is understood about the development of these multifactorial hypertensive disorders, however there is increasing evidence that the manifestation of hypertensive disorders during pregnancy is not solely due to placental-derived dysfunctions and has important maternally-driven components. The stroke-prone spontaneously hypertensive (SHRSP) rat is a well-established model of human essential hypertension. SHRSP dams remain hypertensive throughout pregnancy and demonstrate an abnormal uterine artery structure and function at term. This project aimed to more fully characterise pregnancy in the SHRSP rat. The objectives were to assess maternal responses and determine how maternal hypertension impacts maternal and fetal well-being as well as placental development; to investigate the underlying genetic mechanisms behind the eventual abnormal pregnancy-dependent uterine artery remodelling; and, finally, to increase the maternal cardiovascular load in SHRSP pregnancy to establish a model of superimposed pre-eclampsia. In vivo and ex vivo techniques were used to characterise cardiovascular function in the hypertensive SHRSP and normotensive WKY rats during gestation, as well as assess pregnancy outcomes. The SHRSP dams were found to have similar cardiac function compared to WKY, yet there was evidence of impaired systemic vascular structure and function in late gestation and placental abnormalities. Nevertheless, the SHRSP maintained similar litter sizes to WKY and did not demonstrate any major impact on fetal growth. Further similarities between SHRSP and WKY pregnancy were revealed with the assessment of uterine artery function in early gestation. However, using RNA sequencing to elucidate the transcriptomic profiles of the uterine arteries, SHRSP were found to have strikingly different responses to pregnancy at the transcript expression level, compared to WKY. Finally, a model of superimposed pre-eclampsia was established by increasing the cardiovascular stress in the dam using angiotensin II infusion during pregnancy in SHRSP. This model had a significantly higher systolic and diastolic blood pressure than the already hypertensive SHRSP. The pregnancy-dependent increase in cardiac output, observed in SHRSP, was negated by AngII infusion and was reduced in the highest treatment group. These major cardiovascular impairments were observed alongside increased proteinuria and reduced fetal growth; all phenotypes found in severely pre-eclamptic women. This work has provided information on systemic and uterine specific vascular responses to pregnancy in SHRSP and WKY rats alongside detail of underlying transcriptional differences. This study was the first to examine uterine artery gene expression changes during pregnancy. The different transcriptomic profiles of early pregnancy changes in the two strains make this an intriguing model to study maternal-driven vascular remodelling in hypertensive pregnancy. Furthermore, this work demonstrated that increasing the cardiovascular load during pregnancy in SHRSP successfully mimics superimposed pre-eclamptic phenotypes and could be used in the assessment of novel therapeutic strategies. To conclude, the SHRSP has the potential to aid our understanding of human pre-eclamptic conditions, especially when endeavouring to determine the impact of maternally-driven components of hypertensive disorders of pregnancy.
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11

Spikings, Emma Catherine. "Mitochondrial DNA replication in pre-implantation embryonic development." Thesis, University of Birmingham, 2007. http://etheses.bham.ac.uk//id/eprint/45/.

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All eukaryotic cells possess mitochondrial DNA (mtDNA), which is maternally inherited through the oocyte, its replication being regulated by nuclear-encoded replication factors. It was hypothesised that mtDNA replication is highly regulated in oocytes, pre-implantation embryos and embryonic stem cells (ESCs) and that this may be disrupted following nuclear transfer (NT). MtDNA copy number decreased between 2-cell and 8-cell staged porcine embryos and increased between the morula and expanded blastocyst stages, coinciding with increased expression of mtDNA replication factors. Competent porcine oocytes replicated their mtDNA prior to and during in vitro maturation to produce and maintain the 100000 mtDNA copies required for fertilisation. Those oocytes in which mtDNA replication was delayed had reduced developmental ability. Expression of pluripotency-associated genes decreased as murine ESCs differentiated into embryoid bodies, although expression of mtDNA replication factors did not increase until the stage equivalent to organogenesis. Cross-species NT embryos in which the donor cell-derived mtDNA was replicated produced decreased developmental outcomes compared to those in which no mtDNA replication took place. Disruption of the strict regulation of mtDNA replication that occurs during early embryogenesis, as is likely following NT, may therefore contribute to the reduced developmental ability of embryos produced using such techniques.
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12

Sellayah, Dyan. "Mechanisms underlying developmental induction of energy homeostasis and cardiovascular function in a mouse model : impact of pre- and post-natal nutritional mismatch." Thesis, University of Southampton, 2009. https://eprints.soton.ac.uk/375583/.

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13

Vlachava, Maria. "Salmon In Pregnancy Study (SIPS): the effects of increased oily fish intake during pregnancy on maternal and cord blood fatty acid composition, cord blood immunity and atopy outcomes in infants at 6 months of age." Thesis, University of Southampton, 2010. https://eprints.soton.ac.uk/199377/.

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Parallel increases in many inflammatory diseases including atopy over the last 40 years suggest that common environmental changes may be promoting inflammatory immune responses. Modern diets have become increasingly rich in n-6 polyunsaturated fatty acids (PUFAs) and relatively deficient in n-3 PUFAs. These dietary changes are believed to promote a pro-sensitisation, pro-allergic and pro-inflammatory environment. Exposure to such an environment during pregnancy and in the very early life period is considered to influence subsequent patterns of the immature and developing neonatal immune system, and this may contribute to the increase in allergic disease in early life. As allergic diseases often first manifest in infancy, prevention strategies need to be targeted early, even in utero. Epidemiologic and experimental data provide a plausible link between dietary changes and increased incidence of childhood atopic disease. Although there have been studies examining the potential benefits of giving n-3 PUFA-rich fish oil supplements during pregnancy, there are no studies examining the effects of increased consumption of oily fish in pregnancy on neonatal immune responses and subsequent clinical outcomes. The Salmon in Pregnancy Study (SIPS) is the first randomised controlled trial of oily fish intervention during pregnancy. The hypotheses being investigated in SIPS is that increased intake of salmon, a source of long chain (LC) n-3 PUFAs (eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA)), in pregnancy will a) increase maternal LC n-3 PUFA intake, b) increase maternal and infant blood LC n-3 PUFA status, c) modulate fetal/neonatal immune responses and d) lower the risk of infant atopy determined at 6 months of age. The primary outcome measures of SIPS were the clinical signs of atopy in the offspring. Pregnant women (n=123) at high risk of having atopic offspring, and with low habitual intake of oily fish (≤ 2/month) were randomised at 20 weeks of pregnancy to either consuming 2 portions/week of farmed salmon (n=62) or continuing their habitual diet (n=61) until the end of pregnancy. The woman attended a clinic at 20 (n=123), 34 (n=110) and 38 (n=91) weeks of gestation at which fasting blood was collected and a food frequency questionnaire (FFQ) was administered (at 20 and 34 weeks). At delivery umbilical cord blood was collected (n=101) for fatty acid and immunological analysis. Infants attended a clinic at 6 months of age (n=86) for assessment of allergic sensitisation by skin prick testing (SPT) using various allergen extracts and of atopic dermatitis (SCORAD index). Maternal and cord plasma and cord blood mononuclear cell (CBMC) fatty acid compositions were determined by gas chromatography. Neonatal (cord) immune cell subsets were identified by flow cytometry. Ex-vivo cytokine production by CBMC in response to stimulants (allergen, mitogen, and toll-like receptor (TLR) ligands) was determined by cytometric bead array and flow cytometry. Ex-vivo prostaglandin E2 production by CBMC was determined by enzyme-linked immunosorbent assay. Immunoglobin E concentration was measured in cord blood plasma and in 6 month infant blood plasma. Eating oily fish twice a week during pregnancy resulted in a higher maternal intake of LC n-3 PUFAs (both EPA and DHA) and in higher maternal and cord blood plasma status of LC n-3 PUFAs (both EPA and DHA). LC n-3 PUFA content of CBMC was not significantly affected. CBMC production of interleukins-2, -4, -5, and -10 and tumour necrosis factor-α was lower in the salmon group. There was no effect of salmon on the atopic outcomes assessed at 6 months.
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14

Bradley, Josephine. "Label-free multiphoton microscopy of lipid droplets in oocytes, eggs, and early embryos." Thesis, Cardiff University, 2016. http://orca.cf.ac.uk/97984/.

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Successful development of mammalian oocytes, eggs and embryos relies on the production of ATP by their mitochondria, through metabolism of pyruvate and fatty acids. Imaging of lipid droplets in mammalian eggs has proven difficult due to the invasive, unspecific and unquantitative nature of fluorescent lipophilic stains. Here, we show that coherent anti-Stokes Raman scattering (CARS) microscopy can be used to image lipid droplets in live mouse oocytes and pre-implantation embryos, in a label-free, chemically-specific manner. CARS enables visualisation of lipid droplet distributions, and quantitation of droplet size, number and spatial distribution, notably whilst maintaining their developmental viability. CARS also allows examination of the type of lipids comprising lipid droplets, through means of hyperspectral imaging. It is shown that the chemical composition of these droplets differ in oocytes matured in media supplemented with saturated and unsaturated fatty acids. Correlation of CARS measurements with simultaneous two-photon fluorescence (TPF) microscopy of conventionally used lipid dyes demonstrates only partial correlation, and shows their lack of specificity and unpredictable staining patterns. Dynamic monitoring of lipid metabolism in eggs allows determination of the extent of fatty acid oxidation occurring in mouse oocytes and embryos. Investigation into how inhibition of fatty acid metabolism affects the mitochondrial redox state, membrane potential and ATP level allows further understanding of why fatty acid metabolism is significant for egg and pre-implantation embryo development. Starvation and fatty acid-feeding of oocytes shows that the lipid droplet distribution reflects their level of lipid metabolism, while the detrimental effects of palmitic acid are shown to involve its action at the endoplasmic reticulum SERCA pumps.
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15

Higgins, Claire Angela. "Parturition, oxytocin, inflammation, myocyte damage and obesity : a study of myometrium and haematological parameters in human pregnancy and labour at term." Thesis, University of Glasgow, 2013. http://theses.gla.ac.uk/3804/.

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The process of parturition resulting in the delivery of a newborn is a fundamental event ensuring survival of the species. In humans, the main clinical problems of parturition include activation of the process too early or too late resulting in the delivery of pre-term and post-term infants, both with their own implications for future health for the mother and baby. Additionally, where parturition systems are not activated correctly, dysfunctional labour with the resulting need for caesarean delivery (CS), in addition to atonic post-partum haemorrhage can also ensue. Overall, in the UK up to 40% of pregnancies are affected by one of these problems. However, the exact processes involved in the initiation and maintenance of parturition in the human are not fully understood. With such an important event, influences are most likely to be multi-factorial, with hormonal, mechanical, inflammatory, biochemical and maternal environmental factors playing a part. The aims of this thesis were to investigate influences on parturition in human pregnancy. Firstly, the myometrial transcriptional effects of long term exposure to the uterotonic oxytocin (OT) were examined. Further investigation of the myometrial and maternal peripheral response to uterine contractions in-vitro and in-vivo was also made with particular reference to the role of inflammation and myocyte damage. Additionally, the influence of maternal factors, particularly obesity, on the myometrial in-vitro contractile function and response to OT was studied. Initially, 150 gene arrays were produced using the Illumina platform. The samples were derived from myometrium taken at pre-labour CS which subsequently underwent functional contractility experiments in an organ bath. Five drug environments were studied, namely OT, acetic acid (OT vehicle), ML7(a tocolytic acting via inhibition of myosin light chain kinase), ML7 & OT and finally DMSO (ML7 vehicle). Additionally, five time-points of 0, 1, 2, 4, and 6 hours after drug addition were used, resulting in 5 samples for each drug and time combination. The results indicated that despite a clear enhancement of myometrial contractile activity by OT, this functional response does not appear to be mediated by cellular transcription. However, there was a clear contraction and time dependent transcriptional wave, with overrepresentation of genes associated with inflammation and cellular damage/apoptosis, and down-regulation of pathways concerning cellular metabolism. These findings were confirmed by QPCR on further myometrial samples undergoing additional in-vitro functional studies. In addition to the temporal and contractile association with the inflammatory response, our data suggest inflammation occurs in response to myocyte cellular damage regardless of mode of damage e.g. contractile or chemically induced. This was demonstrated by inflammatory upregulation in myometrium exposed to the tocolytic agents nifedipine and ritodrine, which is not seen in response to ML7. Additionally, the myometrial inflammatory response was enhanced by the infective agent LPS. However, contrary to other proposals, the enhanced inflammatory response of the myometrium did not alter or promote the in-vitro contractile ability of the myometrium or its response to OT. This myometrial transcriptional data therefore suggests that the inflammatory response of labour is associated with contraction, chemical or infection induced myometrial cellular damage, but would not be considered necessary for a contractile response. Our in-vivo study of peripheral changes in the maternal circulation again supported our in-vitro myometrial data. Data showed that the effect of pregnancy at term was limited to increased white cell count driven by a neutrophilia, with no suggestion of leukocyte priming prior to labour. Additionally, term pregnancy is associated with an increase in CRP, an increase in GCSF (corresponding with the neutrophilia) in addition to suppression of the chemokines CCL11 and CCL22. Subsequently, we found that repeated blood samples taken at 2 hourly intervals during term labour induced dramatic changes in inflammatory cells and inflammatory mediators in the maternal circulation. Importantly, these changes occur in a co-ordinated time and contraction dependent manner, with the degree of inflammation associated with the length of time in labour and the degree of myocyte damage as measured by circulating CK and Mb. Our study of the influence of maternal factors on myometrial contractile ability and response to OT examined in-vitro myometrial contractility of 609 myometrial strips from 85 women. We demonstrated that maternal obesity does not impair spontaneous or OT induced myometrial contractions in-vitro. Furthermore, maternal age, ethnicity, parity, previous caesarean delivery,gestation at delivery and birthweight do not influence in-vitro myometrial spontaneous or OT induced contractile activity. This therefore suggests that the observed implication of these maternal and infant factors on parturition in-vivo (high rates of induction of labour, high rates of intrapartum caesarean delivery and post partum haemorrhage) cannot be explained by an effect on myometrial contraction per se. This therefore merits further investigation as to alternative mechanisms to ultimately promote and effective, uncomplicated and safe labour and vaginal delivery for at risk mothers. In summary, this thesis provides evidence that the myometrial contractions of human labour, whether spontaneous or OT induced are capable of inducing a temporal wave of transcriptional changes associated with the processes of inflammation, cellular damage/apoptosis with inhibition of cellular metabolic processes. In addition, maternal peripheral circulating factors mirror the myometrial transcriptional changes. These changes are highly comparable with those seen in response to exercising skeletal muscle, and in this model have been shown to play an important role in muscle repair and remodelling after exercise. Therefore, we would suggest that the inflammatory reaction typically associated with human labour occurs as a non-specific response to contraction induced cellular damage and may play a role in postpartum repair and remodelling of the uterus.
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