Intrauterine Growth Restricted Groups Research Articles

Effect of porcine reproductive and respiratory syndrome virus 2 on tight junction gene expression at the maternal-fetal interface

Understanding why intrauterine growth restricted (IUGR) fetuses are more resilient to transplacental porcine reproductive and respiratory syndrome virus-2 (PRRSV-2) infection compared to normal fetuses may lead to alternative approaches to control PRRS. Our objective was to compare gene expression of a subset of tight junction proteins in the endometrium (END) and placenta (PLC) of i) IUGR vs N-IUGR fetuses, and ii) across disease progression phenotypes following PRRSV-2 infection. In experiment 1, snap frozen END and PLC from fetuses of non-infected control dams (CTRL) and from high viral load viable (HVL-VIA) fetuses, with both groups further classified as either IUGR or non(N)-IUGR based on brain: liver weight ratio were strategically selected from a large challenge trial. In experiment 2, similar tissues were randomly selected from CTRL and from uninfected thymus (UNIF), (HVL-VIA) and HVL meconium-stained in the body (HVL-MEC-B) of PRRSV-infected dams. The expression of claudin (CLDN) 1, 3, 4, 5, 6, 7, 10, tight junction protein 1 (TJP1) and occludin (OCLN) genes were evaluated by PCR. There were no significant group differences between IUGR and N-IUGR groups, regardless of infection status, that explained the resilience of IUGR fetuses. Regarding disease progression, elevated CLDN3 was observed in END of UNIF, CLDN6 expression was lower in PLC when the fetus became infected (HVL-VIA), and CLDN10 elevated in PLC in fetuses showing evidence of compromise (HVL-MEC-B). Lastly, OCLN gene expression was higher in the END and PLC following maternal infection. In conclusion, differences in TJ integrity were mainly observed following PRRSV-2 infection with stepwise changes corresponding with disease progression.

A Two-Week Insulin Infusion in Intrauterine Growth Restricted Fetal Sheep at 75% Gestation Increases Skeletal Myoblast Replication but Did Not Restore Muscle Mass or Increase Fiber Number.

Intrauterine growth restricted (IUGR) fetuses are born with lower skeletal muscle mass, fewer proliferating myoblasts, and fewer myofibers compared to normally growing fetuses. Plasma concentrations of insulin, a myogenic growth factor, are lower in IUGR fetuses. We hypothesized that a two-week insulin infusion at 75% gestation would increase myoblast proliferation and fiber number in IUGR fetal sheep. Catheterized control fetuses received saline (CON-S, n=6), and the IUGR fetuses received either saline (IUGR-S, n=7) or insulin (IUGR-I, 0.014 ± 0.001 units/kg/hr, n=11) for 14 days. Fetal arterial blood gases and plasma amino acid levels were measured. Fetal skeletal muscles (biceps femoris, BF; and flexor digitorum superficialis, FDS) and pancreases were collected at necropsy (126 ± 2 dGA) for immunochemistry analysis, real-time qPCR, or flow cytometry. Insulin concentrations in IUGR-I and IUGR-S were lower vs. CON-S (P ≤ 0.05, group). Fetal arterial PaO2, O2 content, and glucose concentrations were lower in IUGR-I vs. CON-S (P ≤ 0.01) throughout the infusion period. IGF-1 concentrations tended to be higher in IUGR-I vs. IUGR-S (P=0.06), but both were lower vs. CON-S (P ≤ 0.0001, group). More myoblasts were in S/G2 cell cycle stage in IUGR-I vs. both IUGR-S and CON-S (145% and 113%, respectively, P ≤ 0.01). IUGR-I FDS muscle weighed 40% less and had 40% lower fiber number vs. CON-S (P ≤ 0.05) but were not different from IUGR-S. Myonuclear number per fiber and the mRNA expression levels of muscle regulatory factors were not different between groups. While the pancreatic β-cell mass was lower in both IUGR-I and IUGR-S compared to CON-S, the IUGR groups were not different from each other indicating that feedback inhibition by endogenous insulin did not reduce β-cell mass. A two-week insulin infusion at 75% gestation promoted myoblast proliferation in the IUGR fetus but did not increase fiber or myonuclear number. Myoblasts in the IUGR fetus retain the capacity to proliferate in response to mitogenic stimuli, but intrinsic defects in the fetal myoblast by 75% gestation may limit the capacity to restore fiber number.

Third trimester maternal blood and at birth cord blood lipid profile characteristics in pregnant woman with or without fetal growth restriction

Background: To study the lipid profile of 3rd trimester pregnant women and their cord blood with and without intrauterine growth restriction Design: Observational study, Setting: Department of Obstetrics and Neonatology, in a teaching hospital in North India during February 2013 to August 2014.Methods: Third trimester pregnant women and their neonates. Enrolled 250 women were divided in intrauterine growth restricted and control groups. Outcome Measures: Venous blood Lipid levels of 3rd trimester mothers and their neonate at birth.Results: Women of IUGR group had significantly lowered total cholesterol (TC), triglycerides (TG), high density lipoproteins (HDL) and low density lipoproteins (LDL)210.2 (19.8), 221.6 (36.2),65.4(11.7)and 130.7 (20.8)compared to 251(55.3), 234.7(35.6), 70.8(19.9) and 181.3(16.8) mg/dl control group. The TC, TG and LDL levels in cord blood of IUGR group were 93.6(16.5), 50.4 (6.5) and 51.8(12.6) as compared 106.4(17.7), 30.7(4.4) and 55.9(12.1) mg/dl to control group (p value <0.05). HDL levels were significantly lower 15.8(4.6) in IUGR group as compared to 26.5(5.4) in control group. The LDL: HDL and TC: HDL ratio was significantly higher in IUGR group. Maternal weight, gestational age, urban residence, primi parity, birth weight and APGAR score were lower, while maternal age, parity, smoking, blood pressure, cesarean sections (%) and male sex (%) of baby was higher in IUGR group.Conclusions: Lipid profile of mothers of IUGR fetuses had significantly lowered cholesterol levels and their cord blood had shown atherogenic phenotype

Altered molecular signatures during kidney development after intrauterine growth restriction of different origins

This study was performed to identify transcriptional alterations in male intrauterine growth restricted (IUGR) rats during and at the end of nephrogenesis in order to generate hypotheses which molecular mechanisms contribute to adverse kidney programming. IUGR was induced by low protein (LP) diet throughout pregnancy, bilateral uterine vessel ligation (LIG), or intrauterine stress (IUS) by sham operation. Offspring of unimpaired dams served as controls. Significant acute kidney damage was ruled out by negative results for proteins indicative of ER-stress, autophagy, apoptosis, or infiltration with macrophages. Renal gene expression was examined by transcriptome microarrays, demonstrating 53 (LP, n = 12; LIG, n = 32; IUS, n = 9) and 134 (LP, n = 10; LIG, n = 41; IUS, n = 83) differentially expressed transcripts on postnatal days (PND) 1 and 7, respectively. Reduced Pilra (all IUGR groups, PND 7), Nupr1 (LP and LIG, PND 7), and Kap (LIG, PND 1) as well as increased Ccl20, S100a8/a9 (LIG, PND 1), Ifna4, and Ltb4r2 (IUS, PND 7) indicated that inflammation-related molecular dysregulation could be a “common” feature after IUGR of different origins. Network analyses of transcripts and predicted upstream regulators hinted at proinflammatory adaptions mainly in LIG (arachidonic acid-binding, neutrophil aggregation, toll-like-receptor, NF-kappa B, and TNF signaling) and dysregulation of AMPK and PPAR signaling in LP pups. The latter may increase susceptibility towards obesity-associated kidney damage. Western blots of the most prominent predicted upstream regulators confirmed significant dysregulation of RICTOR in LP (PND 7) and LIG pups (PND 1), suggesting that mTOR-related processes could further modulate kidney programming in these groups of IUGR pups.Key messagesInflammation-related transcripts are dysregulated in neonatal IUGR rat kidneys.Upstream analyses indicate renal metabolic dysregulation after low protein diet.RICTOR is dysregulated after low protein diet and uterine vessel ligation.

Apparent diffusion coefficient of different areas of brain in foetuses with intrauterine growth restriction.

PurposeThis study aimed to compare the apparent diffusion coefficient (ADC) values of different brain areas between two groups of intrauterine growth restricted (IUGR) foetuses and control cases.Material and methodsA total of 38 foetuses with IUGR and 18 normal control foetuses with similar gestational age were compared using a 3T magnetic resonance scanner. IUGR cases included 23 foetuses with clinical severity signs (group A) and 15 foetuses without clinical severity signs (group B). ADC values were measured in different brain regions and compared among groups. Foetuses with structural brain abnormalities were excluded from the study.ResultsAll foetuses had normal foetal structural brain anatomy. Head circumference (HC) < 5% was more common in IUGR group A compared to IUGR group B (56.5% vs. 13.3%, p < 0.0001). In comparison to the normal group, the ADC values in IUGR foetuses were significantly lower in cerebellar hemispheres (CH) (1.239 vs. 1.280.5 × 10–3 mm2/s, p = 0.045), thalami (1.205 vs. 1.285 × 10–3 mm2/s, p = 0.031) and caudate nucleus (CN) (1.319 vs. 1.394 × 10–3 mm2/s, p = 0.04). However, there were no significant differences in ADC values between IUGR subtypes. Among all brain regions, pons had the lowest ADC values.ConclusionsADC values of thalami, CN, and CH were significantly lower in IUGR than control foetuses, while there was no significant difference among IUGR groups. Further studies are needed to evaluate the prognostic value of ADC changes in IUGR foetuses.

Cord blood fatty acid-binding protein-4 levels are upregulated at both ends of the birthweight spectrum.

Fatty acid-binding protein-4 (FABP4) is an adipokine associated with obesity and signs of the metabolic syndrome. We aimed to investigate at birth in term neonates with normal and abnormal intrauterine growth concentrations of FABP4 and associate them with various perinatal parameters. Serum cord blood FABP4 levels were prospectively determined by ELISA in 80 singleton term appropriate-for-gestational-age (AGA), intrauterine growth-restricted (IUGR) and large-for-gestational-age (LGA) neonates. Compared to the AGA group, cord blood FABP4 levels were increased in the IUGR and LGA groups. Additionally, they were higher in early-term than full-term neonates. A significant U-shaped correlation was recorded between serum FABP4 levels and birthweight. A significant negative correlation between cord blood FABP4 and gestational age in the whole study population was noted. Cord blood FABP4 levels were significantly higher at the extremes of foetal growth at term and negatively correlated with gestational age, being increased in early-term versus full-term neonates. Further longitudinal studies with larger sample sizes are required to elucidate FABP4 implication in foetal growth and its association with future adverse cardiometabolic outcomes in the offspring.

Prolonged amino acid infusion into intrauterine growth-restricted fetal sheep increases leucine oxidation rates.

Overcoming impaired growth in an intrauterine growth-restricted (IUGR) fetus has potential to improve neonatal morbidity, long-term growth, and metabolic health outcomes. The extent to which fetal anabolic capacity persists as the IUGR condition progresses is not known. We subjected fetal sheep to chronic placental insufficiency and tested whether prolonged amino acid infusion would increase protein accretion in these IUGR fetuses. IUGR fetal sheep were infused for 10 days with either mixed amino acids providing ~2 g·kg-1·day-1 (IUGR-AA) or saline (IUGR-Sal) during late gestation. At the end of the infusion, fetal plasma leucine, isoleucine, lysine, methionine, and arginine concentrations were higher in the IUGR-AA than IUGR-Sal group ( P < 0.05). Fetal plasma glucose, oxygen, insulin, IGF-1, cortisol, and norepinephrine concentrations were similar between IUGR groups, but glucagon concentrations were fourfold higher in the IUGR-AA group ( P < 0.05). Net umbilical amino acid uptake rate did not differ between IUGR groups; thus the total amino acid delivery rate (net umbilical amino acid uptake + infusion rate) was higher in the IUGR-AA than IUGR-Sal group (30 ± 4 vs. 19 ± 1 μmol·kg-1·min-1, P < 0.05). Net umbilical glucose, lactate, and oxygen uptake rates were similar between IUGR groups. Fetal leucine oxidation rate, measured using a leucine tracer, was higher in the IUGR-AA than IUGR-Sal group (2.5 ± 0.3 vs. 1.7 ± 0.3 μmol·kg-1·min-1, P < 0.05). Fetal protein accretion rate was not statistically different between the IUGR groups (1.6 ± 0.4 and 0.8 ± 0.3 μmol·kg-1·min-1 in IUGR-AA and IUGR-Sal, respectively) due to variability in response to amino acids. Prolonged amino acid infusion into IUGR fetal sheep increased leucine oxidation rates with variable anabolic response.

Associations between neural injury markers of intrauterine growth-restricted infants and neurodevelopment at 2 years of age

Objectives: The aim of this study was to evaluate the relationships between brain injury biomarkers in intrauterine growth-restricted (IUGR) infants (S100B and neuron-specific enolase (NSE)) and neurodevelopment at 2 years of age.Methods: This prospective case-control study was a cooperative effort among Spanish Maternal and Child Health Network (Retic SAMID) hospitals. At inclusion, biometry for estimated fetal weight and feto-placental Doppler variables were measured for each infant. Maternal venous blood and fetal umbilical arterial blood samples were collected at the time of delivery and neural injury markers S100B and NSE concentrations were measured. Neurodevelopment was evaluated at 2 years of age using the Bayley Scales of Infant and Toddler Development, third edition (Bayley-III).Results: Fifty six pregnancies were included. Thirty-one infants were classified as IUGR and 25 as non-IUGR. Neurodevelopmental evaluation at 2 years of age indicated that there were no between-group differences for any of the tests. For all patients in both groups, we found statistically significant inverse relationships between the concentrations of NSE in the cord blood and the results of the cognitive test (r = −271, p = .042), fine motor subtest (r = −280, p = .036), and social-emotional test (r = −349, p = .015). We also found statistically significant differences between the concentrations of S100B in the cord blood and the results of the cognitive test (r = −306, p = .022) and expressive communication subtest (r = −304, p = .023). For the IUGR group, we found a significant inverse relationship between the concentrations of S100B in the maternal serum and the results of adaptive behavior test (p < .05). In the non-IUGR group, we found statistically significant inverse relationships between the concentration of NSE in the cord blood and the results of the fine motor subtest (r = −446, p = .025) and social-emotional test (r = −489, p = .021). The difference between the concentration of S100B in the cord blood and the language composite score was also statistically significant (p = .038).Conclusions: At 2 years of age, the concentrations of NSE and S100B were higher in the non-IUGR and IUGR groups with the worst scores for some areas of neurodevelopmental evaluation. The value of these biomarkers for prognostic neurodevelopmental use requires further investigation for both non-IUGR and IUGR infants.

Prenatal Growth Patterns and Birthweight Are Associated With Differential DNA Methylation and Gene Expression of Cardiometabolic Risk Genes in Human Placentas: A Discovery-Based Approach.

Inherent genetic programming and environmental factors affect fetal growth in utero. Epidemiologic data in growth-altered fetuses, either intrauterine growth restricted (IUGR) or large for gestational age (LGA), demonstrate that these newborns are at increased risk of cardiometabolic disease in adulthood. There is growing evidence that the in utero environment leads to epigenetic modification, contributing to eventual risk of developing heart disease or diabetes. In this study, we used reduced representation bisulfite sequencing to examine genome-wide DNA methylation variation in placental samples from offspring born IUGR, LGA, and appropriate for gestational age (AGA) and to identify differential methylation of genes important for conferring risk of cardiometabolic disease. We found that there were distinct methylation signatures for IUGR, LGA, and AGA groups and identified over 500 differentially methylated genes (DMGs) among these group comparisons. Functional and gene network analyses revealed expected relationships of DMGs to placental physiology and transport, but also identified novel pathways with biologic plausibility and potential clinical importance to cardiometabolic disease. Specific loci for DMGs of interest had methylation patterns that were strongly associated with anthropometric presentations. We further validated altered gene expression of these specific DMGs contributing to vascular and metabolic diseases (SLC36A1, PTPRN2, CASZ1, IL10), thereby establishing transcriptional effects toward assigning functional significance. Our results suggest that the gene expression and methylation state of the human placenta are related and sensitive to the intrauterine environment, as it affects fetal growth patterns. We speculate that these observed changes may affect risk for offspring in developing adult cardiometabolic disease.

Medium chain fatty acids in intrauterine growth restricted and small for gestational age pregnancies

Low birth weight is associated with an increased risk of heart disease, high blood pressure and diabetes in adult life. Fetal growth is determined by nutrient availability, which is related to placenta nutrient transport. Medium chain fatty acids (MCFAs) are a particular class of nutrients, known to be a readily available energy source. Until now no data are reported on these MCFAs in low birth weight fetus. This is a prospective study conducted in a tertiary center of prenatal diagnosis to investigate the maternal and fetal MCFAs levels in appropriate for gestational age (AGA), intrauterine growth restricted (IUGR), and small for gestational age (SGA) pregnancies. The plasmatic levels of MCFAs in AGA, IUGR and SGA mother–infant pairs were quantified by gas chromatography–mass spectrometry. The analytical method had a linearity range of 0.1–50 mg/L and a limit of quantification of 0.03 mg/L. Reduced fetal growth was defined as an estimated fetal weight below the 3rd–10th percentile for gestational age, with (IUGR) or without (SGA) fetal Doppler abnormalities. Maternal and fetal MCFAs plasma levels were significantly different among SGA, IUGR and AGA groups. Additionally, the observed MCFAs fetal to maternal ratio is >1 for IUGR group, whilst for SGA and AGA the fetal to maternal ratio is less than one. Changes in MCFAs levels in fetal and maternal plasma are not related to placental functionality or nutrients availability, suggesting the presence of a de novo biosynthesis.

Effect of intrauterine growth pattern on serum visfatin concentrations in full-term infants at birth and at 6 months of life.

Visfatin is a hormone discovered in fat cells and is directly related to diabetes. We aimed to investigate the relationship between intrauterine growth pattern and serum visfatin concentrations in full-term infants at birth and at 6 months of life. Cord blood visfatin concentrations were assessed in 90 full-term neonates enrolled into; Group I: 30 appropriate for gestational age (AGA) neonates to healthy mothers, Group II: 30 intra-uterine growth restricted (IUGR) neonates, 19 were born to mothers with pre-eclampsia, Group III: 30 large for gestational age (LGA) neonates, 16 were infants of diabetic mothers (IDMs). Neonates were followed up at six months of age for visfatin concentrations. Cord blood visfatin concentrations were increased in IUGR compared to AGA group (p = 0.002). Cord blood visfatin concentrations were increased in LGA compared to AGA and IUGR groups (P < 0.001, P < 0.001). Cord blood visfatin concentrations were positively correlated to birth weight in AGA, LGA groups (r = 0.39, p = 0.045, r = 0.449, p = 0.013 respectively). Visfatin concentrations in neonates born to mothers with pre-eclampsia and IDMs were higher than in those born to mothers without pre-eclampsia and to non-diabetic mothers (p = 0.040, p = 0.002 respectively). At six months, serum visfatin concentrations decreased compared to cord blood visfatin concentrations in IUGR and LGA groups (p < 0.001). Levels in LGA were still higher than IUGR (p = 0.004). Serum visfatin concentrations were positively correlated to cord visfatin in IUGR neonates (r = 0.497, p = 0.005). Cord blood visfatin concentrations were increased in LGA and IUGR neonates. At six months, serum visfatin concentrations decreased compared to cord blood visfatin concentrations in LGA and IUGR groups, still higher in the former than the latter.

Implication of the myokine irisin in maternal energy homeostasis in pregnancies with abnormal fetal growth

Objective: To prospectively investigate maternal concentrations of the myokine irisin in large for gestational age (LGA) and intrauterine growth restricted (IUGR) versus appropriate for gestational age (AGA) normal pregnancies and associate them with various perinatal parameters.Methods: Plasma irisin and insulin concentrations were measured by enzyme-linked immunosorbent assay (ELISA) and immunoradiometric assay (IRMA), respectively, in a cohort of 80 mothers delivering LGA (n = 30), IUGR (n = 30) and AGA (n = 20) singleton full-term infants.Results: Maternal irisin concentrations were similar among LGA, IUGR and AGA groups and did not correlate with respective insulin ones or maternal body mass index. In a combined group, maternal irisin concentrations decreased with advancing gestational age (p < 0.001) and were lower in multi-, compared to nulliparous women (p = 0.004). In the IUGR group, maternal irisin concentrations were higher in cases of smoking (p = 0.006).Conclusions: Irisin may not be differentially regulated in insulin resistance-associated pregnancy disorders resulting in fetal macrosomia and IUGR. Maternal irisin down-regulation with advancing gestation could possibly contribute to the observed maternal fat accumulation and progressive insulin resistance towards term. Similarly, lower maternal irisin concentrations in multiparous women may reflect the documented positive association between parity and fat deposition. Irisin up-regulation in cases of smoking may indicate the need for enhanced oxygen consumption to maintain energy production under conditions of hypoxia.

Histomorphometry of umbilical cord and perinatal outcome in intrauterine growth restriction

Objective: To evaluate the histomorphometry of umbilical cord (UC) in intrauterine growth restricted (IUGR) newborns compared to appropriate for gestational age (AGA) newborns, and secondly to assess its impact on the intrapartum and perinatal outcome. Materials and Methods: A prospective observational study was conducted at Obstetrics and Gynecology unit of KLES Dr. Prabhakar Kore Hospital and Medical Research Centre, Belagavi. Study involved total 130 UCs of AGA and IUGR newborns. Tissues were fixed in formalin and paraffin embedded sections were examined. For each UC; UC cross sectional area (CSA), UC diameter and UC circumference parameters were measured under microscope with the help of digital image analyzer. Perinatal outcome like gestational age (GA) at delivery, birth weight, mode of delivery, sex of newborn, neonatal outcome and intrapartum complications were recorded. Independent t test was used to compare means and chi-square test for categorical variables. Results: UC of IUGR newborns had significantly reduced UC area, diameter and circumference as compared to AGA newborns. IUGR newborns were associated with significant difference in GA at delivery, birth weight, neonatal intensive care unit admissions (NICU) and fetal distress. No significant difference was found in mode of delivery, sex of newborn and meconium stained liquor among AGA and IUGR groups. Conclusion: Assessment of CSAs of UC and its components can be used as early screening tool for fetal growth and it may provide useful information about pregnancy at risk and help in averting poor perinatal outcome.

Cord blood galectin-1 and -3 concentrations in term pregnancies with normal restricted and increased fetal growth

To determine levels of galectins (gal)-1 and -3 (implicated in angiogenesis/immunologic mechanisms) in intrauterine growth restricted (IUGR), large (LGA) and appropriate for gestational age (AGA) pregnancies, as these groups differ in fat mass, angiogenic patterns and immune responses. Cord-blood (UC) gal-1 and -3 concentrations were measured in 30 IUGR, 30 LGA and 20 AGA singleton full-term infants and their mothers (MS). IUGR, LGA and AGA groups did not differ in gal-1 and -3 concentrations. UC gal-1 levels were lower when mothers were older [b=-0.651, CI 95% -1.186 (-0.116), P=0.018] and UC gal-3 levels were increased when mothers presented gestational diabetes [b=9.836, CI 95% 3.833- (15.839), P=0.002]. In IUGRs MS gal-3 and in LGAs UC gal-1 were decreased in multiparas [b=-5.372, CI 95% -9.584- (-1.161), P=0.014], and [b=-7.540, CI 95% -14.606- (-0.473), P=0.037], respectively. No correlations were found between MS or UC gal-1 and gal-3 concentrations. Lower UC gal-1 levels, when mothers were older, and increased UC gal-3 levels in cases of gestational diabetes, possibly reflect angiogenic activity. In multiparas, decreased MS gal-3 and UC gal-1 levels in IUGR/LGA, respectively, might imply inflammatory response against immunosuppression expected in subsequent pregnancies, as compared to the first one.

Prenatal Hypoxia Causes Long-Term Alterations in Vascular Endothelin-1 Function in Aged Male, but Not Female, Offspring

Prenatal hypoxia can alter the growth trajectory of the fetus and cause lasting health complications including vascular dysfunction. We hypothesized that offspring that were intrauterine growth restricted (IUGR) because of prenatal hypoxia would exhibit altered vascular endothelin-1 (ET-1) signaling in later life. Isolated mesenteric artery responses to big ET-1 (bET-1) and ET-1 were assessed by using wire myography. Male IUGR offspring had 3-fold greater bET-1-induced vasoconstriction compared with controls (n=7 per group; P<0.001); NO synthase inhibition with L-N(G)-nitro-arginine-methyl ester potentiated bET-1-induced vasoconstriction, albeit this effect was 2-fold greater (P<0.05) in male control compared with IUGR offspring. Vascular responses to bET-1 were similar between female IUGR and control offspring (n=9-11 per group). In the presence of L-N(G)-nitro-arginine-methyl ester, pretreatment with the chymase inhibitor chymostatin, the gelatinase inhibitor GM6001, or the neutral endopeptidase inhibitor thiorphan did not alter responses to bET-1; however, the ET-converting enzyme inhibitor CGS35066 almost completely abolished vascular responses to bET-1 in control and IUGR groups. Systolic blood pressure in IUGR male offspring was more responsive to ET-1 antagonism in vivo compared with controls (-9 versus -4 mm Hg; n=5 per group; P=0.02); no such differences were observed in female offspring (n=5-6 per group). These results demonstrate that vascular ET-1 function is programmed by prenatal hypoxia and provide further insights into the sex differences in the long-term vascular effects of developmental stressors.

Effect of a mixture of bovine milk oligosaccharides, Lactobacillus rhamnosus NCC4007 and long‐chain polyunsaturated fatty acids on catch‐up growth of intra‐uterine growth‐restricted rats

To investigate the effect of a nutritional mixture (bovine milk oligosaccharides, Lactobacillus rhamnosus NCC4007, arachidonic and docosahexaenoic acid) on growth of intrauterine growth-restricted (IUGR) rats. IUGR was induced by maternal food restriction. The offspring (males and females) were assigned to: REF (non-IUGR, no mixture), IUGRc (IUGR, no mixture), or IUGRmx (IUGR, mixture). The mixture was given from day 7 to day 58, when tissues and plasma from half of the animals were collected for hormones, metabolites and microarray analysis. The rest received a high-fat diet (HFD) until day 100. Glucose tolerance was measured at 56 and 98 days, and body fat content at 21, 52 and 97 days. IUGRmx had the greatest growth during lactation, but from day 22 to day 54, both IUGR groups gained less body weight than the REF (P < 0.05). In the short-term (58 days), IUGRmx tended to be longer (P = 0.06) and had less body fat (P = 0.03) than IUGRc. These differences were not seen after HFD. Microarray analysis of hepatic mRNA expression at 58 and 100 days revealed a gender-dependent treatment effect, and expression of genes related to lipid metabolism was the most affected. Twelve of these genes were selected for studying differences in DNA methylation in the promoter region, for some, we observed age- and gender-related differences but none because of treatment. The nutritional intervention promoted catch-up growth and normalized excessive adiposity in IUGR animals at short-term. The benefits did not extend after a period of HFD. IUGR and early diet had gender-dependent effects on hepatic gene expression.

396 Early Endothelium-Dependent Vasodilatory Dysfunction Innormotensive, Intra-Uterine Growth Restricted Rats is Restored by L-Arginine and Inhibition of Arginases

Increasing clinical and experimental evidence demonstrates that fetalprogrammed hypertension in adulthood is associated with altered endotheliumdependent vasodilation. However, it is debated whether decreased endothelium-dependent vasodilation precedes or is a consequence of elevated arterial pressure. We studied endotheliumdependent vasodilating capacity and the L-argininenitric oxide pathway in intra-uterine growthrestricted (IUGR), 5-week-old, male rats, before the development of elevated arterial pressure. We furthermore tested the hypothesis that alteration in endothelial vasodilation may be due to a reduction in L-arginine bioavailability. Methods- Pregnant rats were fed either a control or protein restricted diet during gestation. Endothelium-intact aortic rings of 5-week-old male offspring of the control and IUGR groups were isolated and preconstricted with phenylephrine (PE) to test vascularreactivity to acetylcholine (Ach) and sodium nitroprusside (SNP), respectively endothelium-dependent and -independent vasodilators. Other experiments were conducted with L-arginine supplementation or with S-(2-boronoethyl)-L-cysteine (BEC), a specific endothelial arginase inhibitor. Arginase enzymatic activity was measured in aorta. Results- Systolic blood pressure was not different between groups, and increased later in IUGR rats. While the vasodilatory response to SNP was similar in the two groups, a decreased vasodilatory response to Ach (- 20 %, p< 0.05) was observed in the IUGR group. Moreover, L-arginine supplementation and arginases inhibition restored the altered vasodilating capacity of aortic rings of IUGR animals. Arginase activity is enhanced in aorta of young IUGR rats. Conclusions- Reduced endothelium-dependent vascular relaxation precedes the developmentof fetal programmed hypertension in IUGR rats and involves the L-arginine-NO pathway. L-arginine supplementation and inhibition of arginases restore endothelium-dependent vasodilation in IUGR, prehypertensive rats.

Clara cell protein in full-term pregnancies: The influence of intrauterine growth restriction

Clara cell protein (CC16) is an immunomodulatory/anti-inflammatory broncho-alveolar-derived molecule and a biomarker of pulmonary epithelial cells maturity and alveolo-capillary membrane injury. Intrauterine growth-restricted (IUGR) neonates may present with structural lung immaturity, impaired immunocompetence and increased risk for respiratory infections and chronic obstructive lung disease in later life. To investigate circulating CC16 concentrations in maternal, fetal, and neonatal samples from IUGR and appropriate for gestational age (AGA) pregnancies. Serum CC16 concentrations were determined by EIA in 40 mothers and their 20 IUGR and 20 AGA singleton full-term fetuses-neonates on postnatal days 1 (N1) and 4 (N4). No significant differences in CC16 concentrations were observed between IUGR and AGA groups. In both groups, maternal CC16 concentrations were lower compared to N1 and N4 ones (P < 0.001 in each case). Fetal CC16 concentrations were significantly lower compared to N1 and N4 ones (P < 0.001 in each case). In the AGA group, N1 CC16 concentrations were significantly higher than N4 ones (P < 0.001). Combining groups, N1 CC16 concentrations positively correlated with gestational age (r = 0.364, P = 0.021). Finally, the effect of gender, parity, and maternal age on CC16 concentrations was not significant. The lack of differences in CC16 concentrations between IUGR and AGA groups possibly suggests that the lung immaturity and later respiratory diseases, associated with the former, may not be related to early CC16 deficiency. CC16 concentrations increase with advancing gestational age and peak on the first day of life, possibly indicating a vital role of the protein in fetal lung maturation and extrauterine pulmonary adaptation.

Neonatal Outcomes of Late‐Preterm Birth Associated or Not with Intrauterine Growth Restriction

Objective. To compare neonatal morbidity and mortality between late-preterm intrauterine growth-restricted (IUGR) and appropriate-for-gestational-age (AGA) infants of the comparable gestational ages (GAs). Methods. We retrospectively analyzed neonatal morbidity and mortality of 50 singleton pregnancies involving fetuses with IUGR delivered between 34 and 36 6/7 weeks of GA due to maternal and/or fetal indication. The control group consisted of 36 singleton pregnancies with spontaneous preterm delivery at the same GA, in which the infant was AGA. Categorical data were compared between IUGR and AGA pregnancies by X 2 analysis and Fisher's exact test. Ordinal measures were compared using the Kruskal-Wallis test. Results. The length of stay of newborns in the nursery, as well as the need for and duration of hospitalization in the neonatal intensive care unit, was longer in the group with IUGR. Transient tachypnea of the newborn or apnea rates did not differ significantly between the IUGR and AGA groups. IUGR infants were found to be at a higher risk of intraventricular hemorrhage. No respiratory distress syndrome, pulmonary hemorrhage or bronchopulmonary dysplasia was observed in either group. The frequency of sepsis, thrombocytopenia and hyperbilirubinemia was similar in the two groups. Hypoglycemia was more frequent in the IUGR group. No neonatal death was observed. Conclusion. Our study showed that late-preterm IUGR infants present a significantly higher risk of neonatal complications when compared to late-preterm AGA infants.

Perinatal Changes of Plasma Resistin Concentrations in Pregnancies with Normal and Restricted Fetal Growth

Background:The adipocytokine resistin inhibits adipogenesis and induces insulin resistance. Intrauterine growth-restricted (IUGR) neonates have reduced fat mass and changes of endocrine/metabolic mechanisms, predisposing to insulin resistance and metabolic syndrome in adult life. Objectives: To investigate plasma resistin concentrations in maternal, fetal and neonatal samples from IUGR and appropriate-for-gestational-age (AGA) pregnancies and correlate them with respective insulin concentrations. Methods: Plasma resistin and insulin concentrations were determined in 40 mothers and their 20 IUGR and 20 AGA singleton full-term fetuses and neonates on postnatal day 1 (N1) and day 4 (N4). Results: No significant differences in resistin concentrations were observed between AGA and IUGR groups. In the AGA group, maternal resistin concentrations were significantly lower compared to fetal, N1 and N4 ones (p = 0.003, p = 0.017 and p = 0.039, respectively). Maternal resistin concentrations positively correlated with fetal ones (r = 0.527, p = 0.02). In the IUGR group, maternal resistin concentrations were significantly lower compared to N1 (p < 0.001) and positively correlated with N4 concentrations (r = 0.626, p = 0.007). In both groups, the effect of gender, mode of delivery, parity and adjusted birth weight (customized centiles) on resistin concentrations was not significant. No correlation between resistin and insulin concentrations was documented. Conclusions: Lack of difference in resistin concentrations between IUGR and AGA groups, and lack of correlation between resistin and insulin concentrations as well as customized centiles, possibly suggests that resistin may not be directly involved in the regulation of insulin sensitivity and adipogenesis in the perinatal period. Mode of delivery and parity are not associated with circulating resistin concentrations.