<b>Background</b>: Fetal growth restricted (FGR) infants have increased perinatal morbidity and mortality risks. Standard fetal biometry may identify some FGR babies; amniotic fluid measurement and Doppler assessment of blood vessels provide additional, functional assessments, but are often normal in babies with late-onset FGR who are difficult to diagnose. A marker reflecting nutritional status should help to identify FGR fetuses, enabling effective intervention: increased monitoring or delivery and neonatal management. Arm and/or thigh measurements have previously shown potential in 2D and 3D. Limb fat volume has never been measured and could provide an in utero marker of fetal nutritional status. <b>Aims</b>: 1. To develop an ultrasound scanning protocol to acquire 2D images and 3D volumes of fetal arms and thighs. 2. To develop method(s), suitable for use in clinical practice, to measure fat in these images and volumes. 3. To assess the reproducibility of these methods. 4. To assess the validity of these methods by comparing them with MRI images of fetal limb fat. 5. To use these methods in a healthy population to develop reference ranges. 6. To correlate these methods with validated neonatal measurements, to assess whether the antenatal methods reflect neonatal body composition. 7. To apply these methods to longitudinal prospective ultrasound images from multiple countries to assess SGA fetuses. <b>Method</b>: Ultrasound imaging protocols were developed to obtain accurate images and volumes of fetal arms and thighs. Segmentation tools were designed with biomedical engineers to measure fat, lean and limb compartments, and applied to 2D images and 3D volumes (n=500), with calculation of reference centiles in optimally healthy women (INTERGROWTH-21st study) and MRI validation of the ultrasound measurements. Additional methods were assessed: a two-ellipse method, and a three-thickness Fetal Fat Index (FFI). Reproducibility was assessed using Bland-Altman plots and ICCs. Fetal measurements were correlated with neonatal body composition data and anthropometry. Third trimester fetal thigh fat volumes were compared in sub-cohorts of AGA and SGA fetuses. <b>Results</b>: Reference centiles were calculated for novel fractional arm and leg volumes (fat and lean), from 16 to 41 weeks. 2D reference ranges were also calculated. The FFI technique - quick, simple, 2D - correlated well with fat area and fat volume. DXA analyses showed a strong correlation between neonatal limb and whole body fat. Correlation analyses showed that infants with above- and below-average arm circumferences have significantly different amounts of arm fat as early as 30-35 weeks. The strength of correlation between antenatal limb fat and neonatal PEA POD whole body fat increased with increasing gestational age. Scans at 30-34 weeks showed a significant difference in fractional thigh fat between those who would be born SGA compared with AGA. <b>Conclusion</b>: This thesis explores, in detail, the measurement of fetal arm and thigh fat using 2D and 3D ultrasound, and demonstrates that it is correlated to neonatal body composition thus allowing 'fetal body composition' to be established as a research tool; the ultimate aim is to be able to distinguish growth-restricted fetuses from those of normal nutritional status. Novel measurements have been developed, acquisition protocols described, reproducibility assessed, and reference centiles calculated in an optimally healthy population: 2D Fetal Fat Index, 3D fractional limb fat and lean volumes, and limb fat (2D and 3D) as a percentage.

Fetal growth restriction (FGR) is a common and potentially life-threatening complication that affects 5-10% of human pregnancies. Maternal genetic predisposition and fetal male sex are known risk factors, but the underlying mechanisms are unknown. To study a known maternal genetic risk factor and the impact of fetal sex, we employed a published transgenic (TG) mouse model, which was designed to mimic a common human angiotensinogen (AGT) promoter variant associated with a 20% increase in circulating AGT levels. We hypothesized that TG dams would deliver growth restricted pups and that the underlying mechanism would be related to differences in maternal uterine pregnancy-induced vascular remodeling, abnormal blood flow to the placenta, and placental damage. In addition, since growth restricted human males are at an increased risk of developing adult onset hypertension, which has been associated with reduced nephron development, we tested for developmental programming in our mouse model and the impact of fetal sex. Our results show that TG dams have reduced uterine and placental angiogenesis when their pups were males, but relatively normal angiogenesis in the female siblings compared with wild-type controls. The uterine placental bed in TG dams had abnormal pro-angiogenic/anti-angiogenic expression ratios that were related to differences in uterine natural killer cell activation and fetal sex. The abnormal phenotype could be rescued by delivering vascular endothelial growth factor (VEGF) to uterine endothelial cells. Male progeny from TG dams had abnormal kidney epigenetic changes, fewer nephrons as adults, and they developed stress-induced hypertension. We conclude that the combination of maternal genetic risk and fetal male sex affect uteroplacental angiogenesis leading to FGR and the programming of stress-induced hypertension.

Abstract not available

Intrauterine Growth Restriction (IUGR) is evident in infants born with a reduced weight or length, and/or increased thinness for gestational age. IUGR is associated with altered postnatal growth and regulation, due to unknown mechanisms. Much clinical IUGR results from the reduced delivery of essential substrates (oxygen and nutrients) to the fetus, due to either maternal or placental limitations. Catch-up growth (accelerated rate of growth in absolute or fractional terms) occurs in the majority of IUGR infants, and returns an infant to their predetermined growth curve. IUGR is associated with increased risks of morbidity and mortality in the perinatal period, and with a reduced final adult stature and increased risk of adult onset diseases, particularly diabetes and cardiovascular disease. Catch-up growth after IUGR predicts improved health in terms of reduced hospital visits in infants and children, and an increased final adult stature but also predicts an increased risk of developing obesity, as well as diabetes and cardiovascular disease. The underlying mechanisms for catch-up growth may contribute to this range of outcomes in later life, but are poorly understood. Studies in IUGR infants have demonstrated increased absolute and/or fractional growth rates following birth, termed catch-up growth, in the presence of reduced or normal plasma concentrations of the thyroid hormones and major anabolic hormones (insulin and/or IGF-I). This suggests that increased sensitivity to, rather than increased production of insulin, IGF-I and thyroid hormone, causes catch-up growth following IUGR. We therefore hypothesised that placental restriction of fetal growth would reduce size at birth and increase postnatal growth and adiposity in association with increased metabolic sensitivity to insulin, IGFs and thyroid hormones. This study has shown that the placentally restricted (PR) lamb has a reduced size at birth in terms of soft and skeletal tissues, has increased rates of growth postnatally, and has increased adiposity by six weeks of age. We have also shown that PR of fetal growth in the sheep did not alter gestational age at delivery, but reduced survival rate. PR lambs demonstrated catch-up growth in most parameters by 30 days of age and increased adiposity at six weeks of age compared to the control lambs. Placental restriction increased insulin and IGF sensitivity of circulating free fatty acids, which in turn, predicts increased adiposity. Neonatal catch-up growth after fetal growth restriction was substantially predicted by both abundance of, and metabolic sensitivity to insulin, suggesting increased insulin action as an underlying cause. Catch-up growth occurs in the neonate despite reduced concentrations of fasting plasma IGFs, along with increased IGF sensitivity of free fatty acid metabolism and adiposity. Plasma TH concentrations predicted growth of soft and skeletal tissue in lambs during early postnatal life, particularly in those undergoing catch-up growth following PR. Therefore neonatal catch-up growth after IUGR is associated with increased sensitivity to both insulin and IGFs, particularly of circulating free fatty acids, and appears to occur to the extent allowed by the prevailing abundance of these hormones and of thyroid hormones. If this altered endocrine state persists, increased adiposity and its subsequent amplification may contribute to the development of obesity, and related adverse metabolic and cardiovascular outcomes in adult life. / Thesis (Ph.D.)--School of Molecular and Biomedical Science, 2004.

Low birth weight is associated with an increased risk of impaired glucose tolerance and type 2 diabetes and with signs of increased hypothalamic pituitary adrenal axis activity in later life (1, 2). Low birth usually weight reflects a reduction in fetal growth, which largely depends on an adequate supply of nutrients and oxygen. Variations in supply modify the metabolic and neuroendocrine characteristics of the fetus, which in turn modulate the pattern of functional development as well as growth (3). An adverse fetal environment, evident as low birth weight, is therefore proposed to alter functional development with long term effects for the function and risk of disease in the individual later in life (4, 5). Increased HPAA impairs metabolic homeostasis and could therefore mediate effect of prenatal challenge on later metabolic control (6). It was therefore hypothesised that restriction of fetal growth, increases circulating cortisol and/or alters sensitivity to cortisol, which increases fasting blood glucose, and impairs glucose tolerance in the young adult. Large litter size in the guinea pig is characterised by reduced placental and fetal growth, reduced size at birth and insulin resistance in offspring in later life, providing a suitable model to test this hypothesis. Spontaneous restriction of fetal growth in the guinea pig, evident as small size at birth, was associated with increased salivary cortisol, in both sexes but at different stages of postnatal life. In males, salivary cortisol was increased with small size at birth in early and adult life, but reduced later with ageing. In females however, salivary cortisol was increased in juveniles and in aged adults, possibly reflecting the impact of the oestrus cycle on cortisol production in mature cycling females. Altered activity of the HPGA, which can influence that of the HPAA, has also been reported to be programmed by prenatal restriction. In the guinea pig, salivary testosterone in males increased with age and small size at birth in juveniles, young and aged adults. In females, salivary progesterone increased with age up to 300 days, and decreased with size at birth in the young guinea pig. Although testosterone inhibits HPAA activity, in males, mean salivary cortisol correlated positively with mean salivary testosterone at 100 and 300 days of age. In contrast, progesterone may enhance HPAA activity, and consistent with this, in females, mean salivary progesterone correlated with mean salivary cortisol at 400 days of age. Therefore, salivary testosterone in the male and salivary progesterone in the female guinea pig changes with maturation and has previously reported in this or other species, but small size at birth increases salivary testosterone in males with modest effects in early life in females. This together with the unexpected positive associations of salivary cortisol with testosterone in males, suggests that programming of the HPAA makes little contribution to that of the HPAA as indicated by salivary cortisol. Here we show that low birth weight is associated with increased fasting blood glucose and impaired glucose tolerance in both male and female young adult guinea pigs aged 100 days. Fasting and mean (during IVGTT) plasma cortisol was reduced in low birth weight female adult guinea pigs, and is not vary with size at birth at this age in males. This suggests that circulating cortisol does not contribute to the impaired glycaemia associated with small size at birth in the guinea pig. Glucose tolerance was increasingly impaired in males but not females, as mean plasma cortisol increased. This is consistent with cortisol impairing glycaemia in the guinea pig as in other species, in males at least. To assess the role of cortisol in prentally programmed impairment of glycaemia directly, metyrapone or vehicle containing 24% ethanol was administered to young adult guinea pigs for 3 days. Treatment with the latter impaired fasting blood glucose and glucose tolerance in females and the latter in males compared to a previous IVGTT and this was exacerbated in low birth weight females. Metyrapone prevented this impairment of fasting glycaemia and glucose tolerance in the low birth weight adult female guinea pig and in the male guinea pig regardless of birth weight class. Neither vehicle or metyrapone altered plasma cortisol, before or during a second IVGTT. Limited numbers of animals, particularly females, limited this study however and additional investigation is required. Nevertheless this shows for the first time that inhibition of glucocorticoid synthesis in the guinea pig improves glucose control. Furthermore this suggests that the low birth weight guinea pig may be more sensitive to cortisol, have increased cortisol synthesis or reduced inactivation of cortisol in peripheral tissues, leading to increased local cortisol action. In conclusion, alterations in peripheral HPAA activity in the guinea pig due to restricted fetal growth may contribute to their prenatally programmed development of impaired glucose tolerance as young adults, but the extent of that contribution may vary with age and gender. / Thesis (Ph.D.) -- University of Adelaide, School of Paediatrics and Reproductive Health, 2008

The purpose of this study was to look at the genotoxic and cytotoxic effects of benzo-á-pyrene (BáP), a chemical mutagen that is present in cigarette smoke, on the insulin-like growth factor-I (IGF-I) gene. Women who smoke during pregnancy are more likely to have a growth-restricted baby. We hypothesized that BáP exerts its effects through genotoxic and cytotoxic avenues. The cytotoxicity is manifested by chromosomal abnormalities and a decrease in the rate of cell division. The genotoxicity is manifested by changes in certain genes known to be important in mammalian fetal development such as IGF-I. IGF-I is implicated in intrauterine growth restriction (IUGR), a problem that greatly increases the risk of perinatal morbidity and mortality. To futher understand the mechanism by which BáP influences the normal growth and development of human placental cells, human placental trophoblast cells from an established immortalized cell line were utilized. Cells were cultured in appropriate media, starved (using starvation "Serum Free Medium"), and treated with two doses of BáP, 1µM (dose 1) and 5µM (dose 2). Chromosomes were prepared for cytogenetic analysis and visualized using light microscopy after Giemsa staining. Chromosomal aberrations were identified and the rate of cell division was determined through the analysis of the mitotic index for treated cells compared to a control group. To further understand the influence of BáP on the IGF-I gene expression level, RNA was extracted from control and treated cells, from which cDNA was synthesized and used for further analysis using polymerized chain reaction (PCR). The PCR results were used to better understand the genotoxicity of BáP, while chromosomal aberration analysis was used to determine the cytotoxic effects of BáP on human placental cells. Our results indicate that many chromosomal abnormalities were present in the treated groups compared to the control group. In addition, there was a significant decrease in the mitotic index of the BáP-treated cells (MI=0.3%) verses the control group (MI=0.93%), p value 0.0447. Through the PCR assay, we speculate that there is a dose-related response to BáP of the IGF-I RNA expression level, with low levels in the treated groups compared to the control group. We conclude from these results that BáP influences placental cells at both the gene and chromosome level. It also affects the cell cycle of human placental cells. It is known that smoking is deleterious for fetal development. We believe that the current study brings us closer to understanding the mechanism by which smoking can lead to fetal growth restriction.

This retrospective case-control study was designed to investigate the relationship between placental localization and intrauterine growth restriction (IUGR). Pregnant women with an anatomic survey from January 1, 2000, to December 31, 2005, and delivery of the pregnancy at Yale-New Haven Hospital (YNHH) were identified using clinical and billing records. Multiple gestation, fetal anomaly, and incomplete medical information were reasons for exclusion. Cases (N=69) were consecutive pregnancies with evidence of IUGR (estimated fetal weight <10th percentile for gestational age) at last follow-up ultrasound. Randomly selected controls (N=258) from the same time period had no evidence of IUGR. Maternal, ultrasound, delivery, and perinatal data were collected by retrospective medical record review, and IUGR cases and non-IUGR controls were compared using the Students t-test, Wilcoxon test, Chi-square analysis, Fishers exact test, and ANOVA. Placental location was determined from the anatomic survey record (obtained at 18.4 ± 1.2 weeks gestation in the IUGR group and 18.2 ± 1.0 weeks gestation in the control group; P=0.18). Multivariate logistic regression with adjustment for confounders was used to investigate the association between IUGR and placental localization. Consistent with known predictors of IUGR, the IUGR group had a higher proportion of black women (36.4% vs. 19.8%, P=0.03), chronic hypertension (26.0% vs. 3.5%, P<0.001), and hypertensive disorders of pregnancy (36.2% vs. 5.0%, P<0.001). Mean birth weights of IUGR and non-IUGR pregnancies differed by 2 kilograms (3244 ± 625 grams vs. 1277 ± 637 grams, P<0.001). IUGR infants were more likely to receive antenatal steroids, deliver preterm, deliver by cesarean section, and be admitted to neonatal intensive care. In both IUGR and non-IUGR pregnancies, the placenta was most commonly anterior or posterior. Unilateral placentas were three times more common in the IUGR group than in the non-IUGR group (17.4% vs. 5.0%, P=0.01). IUGR pregnancies were over four times as likely as control subjects to have unilaterally-located placentas compared to anterior placentas (OR 4.8, 95% confidence interval, 1.9-11.7). Adjusting for ethnicity, chronic hypertension, and hypertensive disorders of pregnancy did not affect this finding (OR 4.6, 95% confidence interval 1.6-13.5). In conclusion, we compared a group of 69 IUGR pregnancies to 258 non-IUGR controls and found intrauterine growth restriction to be associated with unilateral placentation.

Thesis (M.Sc.)--University of Alberta, 2010. / A thesis submitted to the Faculty of Graduate Studies and Research in partial fulfillment of the requirements for the degree of Master of Science, Centre for Neuroscience. Title from pdf file main screen (viewed on January 27, 2010). Includes bibliographical references.

Intrauterine growth restriction (IUGR) reduces bone mass by 10-30% and impairs arachidonic (AA) and docosahexaenoic (DHA) acid status in infants. Because AA and DHA enhance neonatal bone mass, the aim of this study was to determine the effects of dietary 0.5% AA and 0.2% DHA (w/w) prior to weaning on bone and growth. 40 guinea pigs were randomized to either a control (C) or low-protein diet (LP) during pregnancy and the C diet or the C diet with AA+DHA during lactation. Measurements included bone mass, metabolism, and strength, and erythrocyte lipid of sows and offspring from birth to 16 wk post-partum. The LP diet induced IUGR, while the AA+DHA increased bone mass by 5-20% in sows and offspring and corrected growth and bone mass in IUGR pups. Thus, AA+DHA provided in lactation rescues the growth trajectory in an IUGR state and is beneficial to maternal and neonatal bone mass.

Diss. (sammanfattning) Uppsala : Uppsala universitet, 2005. / Härtill 4 uppsatser.