Intrauterine Growth Retardation Group Research Articles

Erythrocyte Zinc Protoporphyrin Is Elevated With Prematurity and Fetal Hypoxemia

To examine the utility of red blood cell (RBC) zinc protoporphyrin/heme ratio (ZnPP/H) as an indicator of fetal iron status, because unfavorable neurodevelopmental outcomes have been associated with poor iron status at birth, as indicated by low serum ferritin, and because few reliable indicators of fetal and early neonatal iron status exist. Consecutively studied preterm and term fetuses at delivery included the following groups: (1) control nonhypoxic, (2) fetuses with intrauterine growth retardation (IUGR), and (3) fetuses of insulin-treated mothers (FDM). We hypothesized (1) that rapid growth velocity associated with an accelerated erythropoiesis among normal fetuses will lead to reduced iron delivery to a rapidly expanding RBC mass and higher umbilical cord blood RBC ZnPP/H and (2) that fetuses that are exposed to pathologic hypoxemia will experience an additional increase in erythropoiesis and higher cord ZnPP/H. ZnPP/H was determined on saline-washed cord blood erythrocytes by hematofluorometry and was examined for its relationship with clinical factors and cord blood laboratory measurements indicative of tissue oxygenation (plasma erythropoietin [EPO] and reticulocyte count) and iron status (plasma ferritin and erythrocyte indices). Statistical testing included 1-way analysis of variance, 2-way analysis of variance with covariates, simple linear regression, and multiple regression analysis. Among control group subjects, gestational age at birth was inversely correlated with RBC ZnPP/H and reticulocyte count and positively correlated with ferritin and EPO. Relative to control subjects, IUGR and FDM fetuses at specified gestational age groupings had higher ZnPP/H, lower plasma ferritin, and higher plasma EPO. Statistical modeling of the relationship between ZnPP/H and plasma ferritin among all study groups demonstrated significant impacts of gestational age, plasma EPO, maternal hypertension, and maternal smoking. The inverse association of fetal ZnPP/H with gestational age at birth among control subjects is attributable to erythropoietic stimulation likely as a result of increasing growth velocity at the earliest gestational ages. The relatively higher ZnPP/H observed among fetuses in the IUGR and FDM groups likely is attributable to increased erythropoietic activity secondary to pathologic hypoxemia. Decreased placental iron transfer may also have limited iron availability and contributed to elevated ZnPP/H in the IUGR group. These data support the concept that increased erythropoietic activity and/or limited iron transport may place infants of diabetic mothers and infants with growth retardation at risk for developing systemic iron deficiency later in infancy and in early childhood.

Deficits in spatial orientation of children with intrauterine growth retardation

The spatial orientation of intrauterine growth retarded (IUGR) children versus age-matched controls was examined using two spatial tests. The first test was the radial arm maze (RAM), a navigational test frequently used in animal models. The second test was a subtest from the Kaufman assessment battery for children (K-ABC). The IUGR group comprised 28 children aged 6 years. The control group comprised 29 appropriate-for-gestational age children. The performance of the IUGR children was significantly inferior to controls in both tests. In the RAM test, the ratio between the correct entrances to the total entrances was significantly lower in the IUGR group than in the control group ( P < 0.001). In the K-ABC, the IUGR group could not perform as well as control children ( P < 0.001). These results suggest that spatial orientation in IUGR children is inferior to their age-matched controls, possibly contributing to their potential learning difficulties. The present results also suggest that the RAM can be potentially used to test spatial orientation of children at-risk.

Endocrine Profile of Children with Intrauterine Growth Retardation

Intrauterine growth retardation (IUGR) is a major cause of short stature in childhood. Most but not all children experience catch-up growth by 2 years of age. We investigated the endocrine profile (thyroid function, prolactin, cortisol, C-peptide and insulin-like growth factor-I [IGF-IJ levels) of 57 children with IUGR, aged 2-10 years, and compared it with 30 controls whose birth weight was appropriate-for-gestational-age. The hormonal profile for both groups was similar for thyroid hormones, prolactin, C-peptide and IGF-I. Cortisol levels were significantly lower in the IUGR group compared to controls (p <0,05). When the IUGR group was divided into 'catch-up' growth and 'non-catch-up' subgroups, the latter had significantly lower IGF-I levels (p <0.001). Lower cortisol levels in children born with IUGR may reflect impaired function of the hypothalamic-pituitary-adrenal axis associated with this condition. The significantly lower IGF-I levels of the 'non-catch-up' subgroup may be involved in their failure to grow.

Effect of early nutrition on intestine development of intrauterine growth retardation in rats and its correlation to leptin

To investigate the intestine and body development of intrauterine growth retardation (IUGR) rats under early different protein diet and to analyze the correlation between leptin and intestine and body development. An IUGR rat model was established by food restriction of pregnant female rats. Fifty-six neonatal IUGR rats and 24 neonatal normal rats were randomly divided into normal control group (C group), IUGR model group (SC group), low protein diet IUGR group (SL group), and high protein diet IUGR group (SH group). Eight rats were killed per group at wk 0, 4, and 12. Serum leptin, body weight (BW), body length (BL), intestinal weight (IW), intestinal length (IL), and intestinal disaccharidase (including lactase, maltase, and saccharase) were detected. BW (4.50+/-0.41 g), BL (5.96+/-0.40 cm), IW (0.05+/-0.01 g), and IL (15.9+/-2.8 cm) in neonatal IUGR rats were much lower than those in C group (6.01+/-0.55 g, 6.26+/-0.44 cm, 0.10+/-0.02 g, 21.8+/-2.7 cm, P<0.05), while intestinal lactase and maltase activities were higher than those in C group. SH group showed the fastest catch up growth and their BW, BL, IW, and IL reached the C group level at wk 4. SC group showed relatively slower catch up growth than SH group, and their BW, BL, IW did not reach the C group level at wk 4. SL group did not show intestine and body catch up growth. Intestinal maltase (344+/-33 micromol/(min.g)) and saccharase activities (138+/-32 micromol/(min.g)) in SL group were both markedly lower than those in C group (751+/-102, 258+/-27 micromol/(min.g), P<0.05). There were no significant differences in lactase activities at wk 4 and disaccharidase activities at wk 12 among all groups (P>0.05). The leptin level in SL group (0.58+/-0.12 ng/mL) was the highest in all groups, and much lower in SH group (0.21+/-0.03 ng/mL) than that in any other IUGR groups at wk 4 (P<0.05). Leptin was negatively related to BW (r = -0.556, P = 0.001), IW (r = -0.692, P = 0.001) and IL (r = -0.738, P = 0.000) at wk 4, while no correlation was found at wk 12. High protein diet is a reasonable early nutritional mode to IUGR rats in promoting intestine and body catch up growth.

Importance of Ductus venosus Doppler Assessment for Fetal Outcome in Cases of Intrauterine Growth Restriction

Objective: The measurement and evaluation of ductus venosus (DV) blood flow velocity waveform in high-risk pregnancies has been studied intensively in recent years in order to find a more intermediate signal of fetal compromise. Our objective was to study the fetal outcome of pregnancies with intrauterine growth retardation (IUGR) and normal pulsatility of DV compared to an IUGR group with increased DV pulsatility. Methods: The outcome of 42 fetuses before 32 weeks of gestational age without chromosomal or structural aberrations was analyzed. All fetuses showed IUGR <5th percentile based on placental insufficiency diagnosed by pathologic RI >90th percentile of both maternal uterine arteries. One group (30 fetuses; mean weight 730 g/SD 190 g; mean gestational age 197 days/SD 12 days) showed normal, the other (12 fetuses, mean weight 675 g/SD 179 g; mean gestational age 198 days/SD 12 days) reduced, but neither absent nor reverse DV flow during atrial contraction. All 42 fetuses were delivered by cesarean section because of severe variable or prolonged decelerations. We measured blood flow velocities of the DV in every fetus on an average 3.7 days (range 1–5 days) before cesarean section. Fetal outcome was determined by Apgar scores after 5 and 10 min, arterial pH and base excess; neonatal morbidity was recorded by intensive follow-up. Results: There were no significant differences of pH, umbilical artery base excess, Apgar scores and severe neonatal complications between the two groups. Conclusions: Our own data show no correlation between increased pulsatility in the DV (without absent or reverse flow during atrial contraction) and fetal outcome before 32 gestational weeks, even in cases of severe growth restriction based on placental insufficiency. Therefore in these cases reduced DV flow during atrial contraction should cautiously be interpreted regarding obstetrical decisions.

Contractile Cardiac Activity in Fetuses with Intrauterine Growth Retardation: Correlations between Regional Nonuniformity, Relaxation, and Afterload

Fetuses of 24 women with normal pregnancies and 14 women with fetal intrauterine growth retardation (IUGR) were tested for shape and movement of the inner borders of the left ventricle (LV) by means of echocardiography. The LV contour was divided into 12 segments. The coefficient of variation (CV) of the movements of individual wall segments was calculated to assess regional nonuniformity of contractions of the LV wall. The ratio between the long and short LV axes was lower in the IUGR group (1.12 ± 0.12) than in the control group (1.75 ± 0.16). Nonuniformity of contractions was higher in the IUGR group (CV = 37.8 ± 17.6%) than in the control group (CV = 21.1 ± 9.9%, P < 0.01). The ratio of the early wave to the atrial wave of transmitral flow (E/A ratio) was lower in the IUGR group (0.69 ± 0.08) than in the control group (0.80 ± 0.08). A negative correlation between the CV and the global ejection fraction of the LV was observed in the fetuses of both groups. In addition, a negative correlation was demonstrated for the E/A ratio and the systolic-to-diastolic flow ratio (S/D ratio) in the umbilical artery. Changes in the shape, nonuniformity of contractions, and diastolic function of the heart in IUGR fetuses are considered to be an adaptation to increased afterload as a result of an elevated resistance of placental vessels.

Identification of Macrosomic, Normal and Intrauterine Growth Retarded Neonates Using the Modified Neonatal Growth Assessment Score

Objective: Separation of Macrosomic, normal and intrauterine growth retarded (IUGR) neonates using the modified Neonatal Growth Assessment Score (NGAS). Methods: A sample of 23 neonates with evidence of growth acceleration and 74 neonates classified as normal or IUGR in a previous study was used in this investigation. The prenatal growth of all neonates had been studied longitudinally with ultrasound. From the data collected, Rossavik models describing the growth trajectories of head circumference (HC), abdominal circumference (AC), thigh circumference (ThC), femur diaphysis length, head cube (A) and abdominal cube (B) were specified. These models were used to predict birth weight (WT), crown-heel length (CHL), HC, AC and ThC at birth. Actual birth measurements of WT, CHL, HC, AC and ThC were evaluated using age-specific normal size curves and compared to predicted measurements by means of the growth potential realization index (GPRI). GPRI values were evaluated by comparison to previously established normal ranges and used to calculate Neonatal Growth Assessment Scores (NGAS₅). NGAS₅ values, together with assessments of anatomical measurements and GPRI values, were used to establish macrosomic, normal and IUGR groups. Principal components analysis was applied to the sets of GPRI values available for each neonate to provide a principal component score for separating macrosomic and normal neonates (m₂NGAS₅₁) or one to separate macrosomic, normal and IUGR neonates (m₃NGAS₅₁) using linear discriminant analysis. The groups identified by these multivariate methods were compared to the original classification and their characteristics evaluated. Results: The m₂NGAS₅₁ and a boundary value of 207.5% separated macrosomic and normal neonates with an accuracy of 97.3%. The m₃NGAS₅₁ and boundary values of 210.0 and 182.5% separated macrosomic, normal and IUGR neonates with an accuracy of 96.9%. No single GPRI value or anatomical measurement could achieve these levels of accuracy. All normal infants were AGA but only 45.5% of the IUGR group were SGA and only 60.9% of the macrosomic group were LGA. Thirteen different types of IUGR and eleven different types of macrosomia were identified based on GPRI values. Conclusion: The modified NGAS accurately separates macrosomic, normal and IUGR neonates although growth abnormalities are expressed in different ways in different individuals.

Localization of neural cell adhesion molecule and pan-cadherin immunoreactivity in intrauterine growth-retarded newborn rat kidneys.

To examine the expression and distribution pattern of neural cell adhesion molecule (N-CAM) and pan-cadherin immunoreactivity in intrauterine growth retardation (IUGR) newborn kidneys, we used a rat model of maternal protein restriction throughout pregnancy. Weak or moderate immunoreactivity for N-CAM and pan-cadherin was seen in some proximal tubules, in the thick segments of Henle, and in the collecting tubules of the control sections. However, the number of tubules expressing N-CAM and pan-cadherin was increased in the IUGR group compared with the control group. Increased density of N-CAM and pan-cadherin immunoreactivity was observed mostly in the proximal tubules, in the thick segments of Henle, and in the collecting tubules of IUGR newborn rat kidneys. Furthermore, N-CAM and pan-cadherin immunoreactivity was present in the thin limb of Henle in the IUGR group, whereas it was absent in the control group. Glomeruli were negative in both groups except for some glomeruli that showed very weak N-CAM staining in the IUGR group. Thus it was demonstrated for the first time that IUGR newborn rat kidneys express N-CAM and cadherin adhesion molecules at specific sites of the nephron.

Persistent short stature, other potential outcomes, and the effect of growth hormone treatment in children who are born small for gestational age.

Persistent short stature, other potential outcomes, and the effect of growth hormone treatment in children who are born small for gestational age.

Placental isoferritin action in pathogenesis of pre-eclampsia and/or intrauterine growth retardation and its earlier predictive value

In order to investigate the role of placental isoferritin (PLF) in pathogenesis of preeclampsia and/or intrauterine growth retardation (IUGR) and its earlier predictive value, a prospective double-blinded study was performed. In 120 initial normal pregnant women at earlier third trimester (from 24 to 34 weeks), plasma placental isoferritin and nitric oxide (NO) metabolites (nitrite/nitrate) (NO2−/NO3−) were examined by using ELISA and Criess assay respectively. The out-come of pregnancies and birth weight of their infants were followed up. The receiver operating characteristic curves (ROC) and predictive values of PLF predicting the outcome of pregnancy with IUGR, pre-eclampsia were analyzed. Results showed that in 120 initial normal pregnant women, IUGR occurred in 15 pregnant women (IUGR group) and pre-eclampsia in 19 (pre-eclampsia group), and the remaining 86 had normal pregnancy (normal group). The levels of plasma placental isoferritin were significantly decreased in IUGR group (260.01±58.95) μg/ml and pre-eclampsia group (285.31±53.73) μg/ml as compared with those in normal group (775.62±89.32) μg/ml at earlier third trimester (bothP 0.05). The plasma placental isoferritin was negatively correlated with NO levels (r=0.329,P<0.01). The areas under ROC of PLF predicting IUGR and pre-eclampsia were 0.977 and 0.905 respectively. At the cut point of 400 μg/ml PLF level, the sensitivity, specificity, positive predictive value, negative predictive value and Kappa index of PLF levels predicting the outcome of pregnancy with pre-eclampsia were 100%, 85.15%, 55.88%, 100% and 0.645 respectively. At the cut point of 390 μg/ml PLF level, the sensitivity, specificity, positive predictive value, negative predictive value and Kappa index of PLF levels predicting the outcome of pregnancy with IUGR were 100%, 81.9%, 44.12%, 100% and 0.663 respectively. It was concluded that the decrease of plasma placental isoferritin levels at earlier third trimester was associated with IUGR and/or pre-eclampsia, and the endothelial cell damage may be one of its mechanisms. The plasma PLF level can be used as an earlier predictor for screening of IUGR and/or pre-eclampsia.

Expression of IGR-IR and VEGF and Trophoblastic Proliferative Activity in Placentas from Pregnancies Complicated by IUGR

Intrauterine growth retardation (IUGR) is recognized as an important cause of low birth weight and elective preterm delivery. IUGR is associated with multiple causative factors, including placental dysfunction. The aim of this prospective study was to investigate the role of trophoblastic proliferative activity and type I insuline-like growth factor receptor (IGF-IR) and vascular endothelial growth factor (VEGF) expressions in the pathogenesis of IUGR. Immunohistochemistry using VEGF, IGF-IR, and Ki-67 antibodies was performed on formalin-fixed placental tissues of third-trimester pregnancies complicated by IUGR (n = 19) and pregnancies with appropriately grown fetuses (n = 27). In addition, histopathological examination of the placentas was performed, and histological findings were categorized into three groups: utero-placental vascular pathologies (UPVP), coagulation-related pathologies, and chronic inflammation. Statistical analysis revealed that villous trophoblastic IGF-IR immunostaining was significantly weaker in placentas with IUGR (p < 0.001), whereas trophoblastic Ki-67 proliferative index and VEGF immunoscoring did not show any significant difference. Histologically, UPVP and chronic inflammation were significant findings in placentas with IUGR (p = 0.04 and p = 0.04, respectively). In addition, placentas were significantly smaller in the IUGR group (p < 0.001). We conclude that villous trophoblastic IGF-IR expression may play a significant role in the pathogenesis of IUGR, and histopathological examination of placentas in pregnancies complicated by IUGR may yield significant findings. In contrast, based on our findings, trophoblastic proliferation and VEGF expression are unlikely to be significant parameters in the pathogenesis of IUGR.

IGFs and Binding Proteins in Short Children with Intrauterine Growth Retardation

The aim of this study was to examine the relationship between the IGF-IGF binding protein (IGFBP) axis and insulin secretion in short intrauterine growth retardation (IUGR) children. Fifteen IUGR and 12 normal short prepubertal subjects had a 90-min frequently sampled iv glucose tolerance test performed to measure plasma glucose, insulin, IGF-I, IGF-II, IGFBP-3, and IGFBP-1. In addition, 29 nonobese prepubertal subjects of normal height had fasting plasma IGF-I and IGFBP-3 levels measured. In comparison to short normal subjects, IUGR subjects had higher plasma values for IGF-I (42 ± 10 vs. 77 ± 31 μg/liter; P < 0.0001), IGF-II (291 ± 76 vs. 370 ± 66 μg/liter; P < 0.008), IGFBP-3 (1.66 ± 0.28 vs. 2.07 ± 0.48 mg/liter; P < 0.0005), fasting insulin (2 ± 1 vs. 4 ± 2 mU/liter; P < 0.004), and acute insulin response (AIR; 215 ± 36 vs. 504 ± 90 mU/liter; P = 0.008). Nonobese subjects of normal height had higher plasma IGF-I (117 ± 9 μg/liter; P < 0.0001) and IGFBP-3 (2.34 ± 0.12 mg/liter) values than the IUGR group (P < 0.0005). During the frequently sampled iv glucose tolerance test, the magnitude of the AIR in short normal subjects was related to the fall in IGFBP-1 levels (P = 0.03); however, no relationship was seen between AIR and fall in IGFBP-1 in IUGR subjects (P = 0.24). In conclusion, short IUGR children have higher plasma IGF-I, IGF-II, and IGFBP-3, when compared with normal children matched for height, weight, and pubertal status. We speculate that hyperinsulinism secondary to insulin resistance may have led to these changes to the IGF-IGFBP axis in the IUGR group.

IGFs and binding proteins in short children with intrauterine growth retardation.

The aim of this study was to examine the relationship between the IGF-IGF binding protein (IGFBP) axis and insulin secretion in short intrauterine growth retardation (IUGR) children. Fifteen IUGR and 12 normal short prepubertal subjects had a 90-min frequently sampled iv glucose tolerance test performed to measure plasma glucose, insulin, IGF-I, IGF-II, IGFBP-3, and IGFBP-1. In addition, 29 nonobese prepubertal subjects of normal height had fasting plasma IGF-I and IGFBP-3 levels measured. In comparison to short normal subjects, IUGR subjects had higher plasma values for IGF-I (42 +/- 10 vs. 77 +/- 31 microg/liter; P < 0.0001), IGF-II (291 +/- 76 vs. 370 +/- 66 microg/liter; P < 0.008), IGFBP-3 (1.66 +/- 0.28 vs. 2.07 +/- 0.48 mg/liter; P < 0.0005), fasting insulin (2 +/- 1 vs. 4 +/- 2 mU/liter; P < 0.004), and acute insulin response (AIR; 215 +/- 36 vs. 504 +/- 90 mU/liter; P = 0.008). Nonobese subjects of normal height had higher plasma IGF-I (117 +/- 9 microg/liter; P < 0.0001) and IGFBP-3 (2.34 +/- 0.12 mg/liter) values than the IUGR group (P < 0.0005). During the frequently sampled iv glucose tolerance test, the magnitude of the AIR in short normal subjects was related to the fall in IGFBP-1 levels (P = 0.03); however, no relationship was seen between AIR and fall in IGFBP-1 in IUGR subjects (P = 0.24). In conclusion, short IUGR children have higher plasma IGF-I, IGF-II, and IGFBP-3, when compared with normal children matched for height, weight, and pubertal status. We speculate that hyperinsulinism secondary to insulin resistance may have led to these changes to the IGF-IGFBP axis in the IUGR group.

Insulin-like growth factor I, epidermal growth factor and transforming growth factor beta expression and their association with intrauterine fetal growth retardation, such as development during human pregnancy.

Fetal intrauterine growth retardation (IUGR) is one of the most common obstetric problems, with a frequency of 12% in Mexico. In the past, investigations have focused on extrinsic causes of IUGR. More recent studies have examined the intrinsic factors that cause fetal intrauterine growth. Maintenance of fetal growth has been attributed to insulin-like growth factor (IGF), epidermal growth factor (EGF) and transforming growth factor beta (TGF-beta). The objective of this study was to assess the levels of these growth factors during pregnancy and to determine whether or not low concentrations are associated with IUGR. Nine women whose pregnancies were complicated by IUGR and a group of nine women whose pregnancies exhibited normal fetal intrauterine growth were studied. IUGR was determined by sonography and confirmed by weight at birth. Venous blood samples were taken from both groups of pregnant women at the end of each trimester. Enzyme-linked immunosorbent assays, immunoradiometric assays and radioimmunoassays were used to process samples, and the results were analysed by anova. IGF-I levels increased in both groups during pregnancy, but the increase was lower (p < 0.001) in the IUGR group throughout pregnancy and at delivery. EGF did not show any significant changes during pregnancy. Blood TGF-beta levels varied only during the first trimester of pregnancy. The differences were not statistically significant. However, TGF-beta concentrations were higher in the pregnancies with IUGR. Women in the IUGR group were smaller than in the control group (p < 0.05), and, using the covariance test (p < 0.05), this was found to be correlated with IGF-I levels but not with EGF or TGF-beta levels. Changes in fetal weight might be explained by the different concentrations of IGF. The structural homology between IGF-1 and insulin could mean that the presence of higher levels of IGF would result in a increased energetic metabolism that could contribute to fetal growth. EGF levels were not related to IUGR, and TGF-beta levels increased only during the first 3 months in the IUGR group. This observation correlates with the in vitro action of TGF-beta as a negative factor of growth, but as a positive support for sustaining early pregnancy. Our data illustrates that low height represents an increased risk factor for IUGR. These data also correlate with the studies involving extrinsic factors.

Circadian rhythm of ANP, aldosterone and PRA in normotensive IUGR.

Atrial natriuretic peptide (ANP) increases are reported during normal pregnancy, but the relation to arterial pressure and the renin-angiotensin system is debatable. We assessed whether normotensive pregnancies with intrauterine growth retardation (IUGR) present an alteration of maternal ANP levels. A total of 11 pregnant women with IUGR, in the absence of any other maternal or fetal pathology, entered the study during the third trimester. They were compared with 12 healthy pregnant women of similar age and characteristics. We monitored all subjects for blood pressure (BP), ANP, aldosterone and plasma renin activity (PRA), under the same conditions for 24 h. All subjects were submitted to the same regimen of life; with homogeneous dark : light periods, salt intake and meal times. BP was monitored at 20 min intervals for 24 h and blood tests performed at six time points during the 24 h. EDTA plasma samples were immediately centrifuged. Hormone assays were performed by radioimmunoassay. Koch's nonparametric two-way analysis of variance (ANOVA) was used to compare the hormone time-dependent profiles in the two groups. Circadian rhythms were assessed by cosinor analysis. The IUGR group was characterized by higher ANP values compared to normal pregnancy, (205 +/- 24 versus 146 +/- 21 pg/ml: P < 0.05) but not significant differences were shown for PRA, aldosterone and BP circadian rhythms. This study shows higher ANP values in human pregnancy complicated by IUGR, with presence of normal BP, aldosterone and PRA profiles.

Decreased galanin mRNA levels in growth hormone-releasing hormone neurons after perinatally induced growth retardation.

Intrauterine growth retardation (IUGR) is associated with persistent postnatal growth retardation accompanied by dysfunction of the hypothalamic components of the growth hormone (GH) axis. At the adult stage, this is reflected by increased somatostatin (SS) and decreased neuropeptide Y (NPY) mRNA levels, whereas the GH-releasing hormone (GHRH) mRNA levels are normal and the output of GH remains unchanged. To extend our insight into the hypothalamic control of GH secretion in growth retarded rats, we determined galanin (GAL) mRNA levels at the adult stage of perinatally malnourished (i.e. IUGR and early postnatally food restricted) rats. Analyses included comparison of GAL mRNA levels in GHRH neurons in perinatally malnourished adult rats using a semi-quantitative double labeling in situ hybridization technique. We report that IUGR is accompanied by a 60% decrease in GAL mRNA levels in all GHRH neurons in the male IUGR group whereas a tendency towards a decrease was observed in the male early postnatally food restricted (FR) group. These effects became more pronounced when the analysis was restricted to GHRH neurons coexpressing GAL mRNA i.e. decreased GAL mRNA levels were seen in both male and female IUGR rats and in FR males. These data show that GAL mRNA levels in GHRH neurons are persistently decreased after perinatal malnutrition. Taking these results together with our previous data on SS, NPY and GHRH mRNA levels, we can conclude that IUGR leads to a reprogramming of the hypothalamic regulation of GH secretion.

Impaired regulation of glucose transporter 4 gene expression in insulin resistance associated with in utero undernutrition.

The aim of this study was to investigate whether insulin resistance-associated in utero undernutrition was related to changes in insulin action on gene expression of molecules involved in the insulin signaling pathway and peripheral glucose metabolism in muscle and adipose tissue. Thirteen insulin-resistant subjects born with intrauterine growth retardation (IUGR) were matched for age, gender, and body mass index to 13 controls. Gene expression of insulin receptor, insulin receptor substrate-1, p85alpha phosphatidylinositol 3-kinase, glucose transporter-4 (GLUT4), hexokinase II, and glycogen synthase was studied in skeletal muscle at baseline and after a 3-h euglycemic insulin stimulation. Target messenger ribonucleic acid (mRNA) levels were quantified using the RT-competitive PCR method. Insulin-stimulated glucose uptake was significantly lower in IUGR-born subjects than in controls (36.9 +/- 12, 7 vs. 53.9 +/- 12.7 micromol/kg.min; P = 0.007), affecting both the glucose oxidation rate and the nonoxidative glucose disposal rate. At baseline, the expression of the six genes in muscle did not significantly differ between the two groups. The insulin-induced changes over baseline were comparable in both groups for all mRNAs, except GLUT4. In contrast to what observed in the control group (mean increment, 49 +/- 23%; P = 0.0009), GLUT4 expression was not stimulated by insulin in the IUGR group (8 +/- 8%; P = 0.42). Moreover, the magnitude of the defect in GLUT4 mRNA regulation by insulin was correlated to the degree of insulin resistance (r = 0.73; P = 0.01). A similar lack of significant GLUT4 mRNA stimulation by insulin was observed in the adipose tissue of IUGR-born subjects. In conclusion, insulin resistance in IUGR-born subjects is associated with an impaired regulation of GLUT4 expression by insulin in muscle and adipose tissue. Our data provide additional information about the mechanism of insulin resistance associated with in utero undernutrition and strengthen the role of glucose transport in the control of insulin sensitivity.

Relatively low serum leptin levels in adults born with intra-uterine growth retardation

In-utero under-nutrition dramatically alters the development of adipose tissue, during the fetal and the neonatal period. To investigate whether adults born with intra-uterine growth retardation (IUGR) show evidence of impaired adipose tissue development and leptin regulation. Serum leptin concentrations were measured in 26 healthy adults born with IUGR and 25 controls aged 24 y who have been studied previously, 3 y ago. The IUGR group demonstrated a significant increase of body mass index (BMI) in comparison to controls between 21 and 24 y of age (4.8+/-7.7%, P=0.004 vs 0.8+/-6.7%, P=0.70). Percentage of total body fat mass was significantly higher in IUGR-born subjects than in controls (27.2+/-7.6 vs 22.0+/-7.3%, P=0.02). Fasting insulin was significantly higher in the IUGR group (7.5+/-3.8 vs 5.3+/-2.3 miccroU/ml, P=0.03). Surprisingly, crude serum leptin concentrations did not significantly differ between the two groups. Moreover, adjusted means of serum leptin levels were significantly lower in IUGR-born subjects than in controls when corrected for body fat mass, gender and fasting insulin (11.3 vs 13.8 ng/ml, P=0.02). Adults born with IUGR developed an excess of adipose tissue associated with relatively low serum leptin levels suggestive of an altered adipocyte function. Considering the close relationship between adipose tissue and insulin-sensitivity, these observations point to the potential implication of abnormal adipose tissue development in the long-term metabolic consequences associated with in-utero undernutrition.

No evidence for a major beta-cell dysfunction in young adults born with intra-uterine growth retardation.

The association between low birth weight and the later development of type 2 diabetes is well established. It has been hypothesized that in utero undernutrition may affect pancreatic beta-cell development, leading to an impaired beta-cell function in adulthood. We have previously demonstrated that intrauterine growth retardation (IUGR) is associated with insulin-resistance early in adulthood. The aim of this study was to test whether IUGR would affect beta-cell function in young adults. Twelve 25-yr-old insulin-resistant subjects with IUGR were matched for age gender and body mass index (BMI) to 13 controls. All of them had normal glucose tolerance. Mean fasting plasma glucose did not significantly differ between the group with IUGR and the control group (97 +/- 7 vs. 98 +/- 4 mg/dL; p = 0.83). Blood glucose was maintained at 124.8 +/- 6.5 vs. 126.2 +/- 7.5 mg/dL above the basal glycemia in the IUGR and control groups (p = 0.64) throughout the hyperglycemic clamp. Serum insulin concentrations did not significantly differ between the group with IUGR and the control group either during the first (0-10 min) phase (311 +/- 252 vs. 248 +/- 184 pmol/L, p = 0.85) or during the second (80-100 min) phase (575 +/- 284 vs. 559 +/- 413 pmol/L, p = 0.72). C-peptide concentrations were similar in both groups during the two phases (2.35 +/- 1.44 vs. 2.59 +/- 1.10 nmol/L, p = 0.39 and 4.86 +/- 1.36 vs. 4.96 +/- 1.41 nmol/L, p = 0.91). In conclusion, our data do not argue in favor of an impairment of beta-cell function at 25 yr of age as a consequence of in utero undernutrition, but rather suggest that insulin resistance might be the primary defect responsible for the development of metabolic disorders associated with IUGR in adulthood.

Telomerase Activity and Apoptosis as Indicators of Ageing in Placenta with and without Intrauterine Growth Retardation

Telomerase activity (TA) and apoptosis were analysed in placenta with and without intrauterine growth retardation (IUGR). Thirty-one specimens were taken from chorionic villi in the first trimester (Group A), 32 placenta specimens were obtained from cases without IUGR in second and third trimester (Group B) and 12 specimens of placenta tissue were obtained from cases of asymmetric IUGR between 26 and 39 weeks of gestation (Group C). TA was analysed by the Telomeric Repeat Amplification Protocol (TRAP) and in situ TRAP assay. Apoptotic changes were assessed by immunohistochemical staining using Bcl-2 monoclonal antibody and by terminal transferase-mediated in situ end-labelling (TUNEL). TA was detected in 29 of the 31 (93.5 per cent) chorionic villi (Group A) and in 20 of the 32 (62.5 per cent) placenta without IUGR (Group B), whereas weak TA was observed in the placenta of all 12 asymmetric IUGR cases (Group C). Significantly higher Bcl-2 immunoreactivity was observed in trophoblastic cells of Group A (85.9±4.2 per cent) and Group B (72.8±7.2 per cent) than Group C (54.84±4.83 per cent), while TUNEL positive cells were identified at a significantly higher frequency in the trophoblastic cells of Group C (9.7±7.4 per cent) than Group A (1.1±1.9 per cent) or Group B (2.9±3.7 per cent).