Evidence for a suppression of apoptosis in early postnatal life.

BACKGROUND: In fetal life the rates of proliferation and apoptosis are high. We studied if the rate of apoptosis decreases after birth, by measuring the soluble (s) forms of Fas-Fas ligand and Tumor necrosis factor receptor 1 (TNFR1)-TNF-alpha which attenuate apoptosis. SUBJECTS AND METHODS: Blood was drawn from the umbilical cord and from a peripheral vein on day 1 and 4 of life in 35 infants and sFas, sFas ligand, sTNFR1 and sTNF-alpha were determined by enzyme immunoassays. Data were analyzed by paired t-test and Pearson's correlation analysis. RESULTS: sFas and sTNF increased significantly from birth to day 4 of life (p=0.033 and p<0.0001 respectively), while sTNFR1 increased significantly from birth to day 1 of life (p<0.0001) followed by a significant decrease on day 4 of life (p<0.0001). The levels of sFas and sTNF-alpha on day 1 correlated with those on day 4 (r=0.677, p<0.0002 and r=0.525, p<0.007 respectively). sFAS ligand was not detectable in any specimen. CONCLUSION: The increasing concentrations of the soluble molecules, sFas, sTNF-alpha and sTNFR1 might indicate that apoptosis may be downregulated in early postnatal life.

Risk factors predisposing to fetal loss following a second trimester amniocentesis.

OBJECTIVE: To examine the influence of possible risk factors on fetal loss rate following amniocentesis. DESIGN: Retrospective analysis of case records between 1993 and 1998. SETTING: Fetal medicine unit of a large teaching hospital. POPULATION: One thousand and six women with singleton pregnancies formed the study group. Seven hundred and eight of them had bleeding during the current pregnancy before the procedure. while 298 had a history of three or more first trimester abortions and/or a second trimester miscarriage or termination of pregnancy. Four thousand and twenty-four women who had amniocentesis and had no risk factors served as controls. Both groups were also classified according to maternal age. Group 1:1,610 women aged 20-34 years; Group 2: 2850 women aged 35-39 years; Group 3; 570 women > 40 years. METHODS: Women of both groups underwent a second trimester amniocentesis between 16 and 21 weeks of gestation. Fetal losses following amniocentesis were examined in three time intervals: 1. in the first two weeks after the procedure; 2. up to the 28th week; 3. from the 28th week to term. RESULTS: There was a statistically significant difference in the fetal loss rate between women aged 20-34 years (2.54%) and those > 40 years (5.1%). Women with a history of vaginal bleeding during the current pregnancy had a higher fetal loss rate compared with controls (6.5% vs 2.8%), which corresponds to an odds ratio of 2.4 (95% CI 1.69-3.42). A similar difference was found between the group of women with a history of previous abortions/terminations and the controls (8% vs 2.8%): OR 3.03 (95% Cl 1.92-4.79). CONCLUSIONS: There is a higher risk of fetal loss following amniocentesis in women > 40 years of age compared with those aged 20-34 years. Bleeding in the current pregnancy, a history of three or more first trimester abortions, a second trimester miscarriage or termination of pregnancy seem to be significant predisposing factors for fetal loss after an amniocentesis.

Gene analysis of the N-terminal region of the estrogen receptor alpha in preeclampsia.

Alterations in the NH(2)-terminal region of the estrogen receptor alpha (ERalpha) gene expressed in placental bed tissue may be implicated in the development of preeclampsia, the pathogenesis of which involves the spiral arteries. Therefore, mutations and polymorphisms on exons 1 and 2 of the gene encoding ERalpha were studied. Placental bed biopsies were taken from 20 healthy, normotensive pregnant women and 16 preeclamptic patients. DNA was extracted from the tissue and exon 1 and exon 2 were amplified by PCR prior to denaturing gradient gel electrophoresis analysis or to single stranded conformational polymorphism analysis. In exon 1, a codon 10 polymorphism, either homozygous for the wild type gene, homozygous for the mutant type gene, or heterozygous, was revealed in both patients and healthy individuals. A codon 87 polymorphism, homozygous for the wild type gene, was detected in both groups. No mutations or polymorphisms were found in exon 2. The allele distribution for either codon 10 or 87 between patients and healthy individuals showed no significant differences. In conclusion, genetic alterations in the NH(2)-terminal region of the ERalpha molecule are not correlated with preeclampsia.

Concentrations of angiogenic factors in follicular fluid and oocyte-cumulus complex culture medium from women undergoing in vitro fertilization: association with oocyte maturity and fertilization.

OBJECTIVE: To determine the concentration of angiogenic factors (vascular endothelial growth factor [VEGF], basic fibroblast growth factor [bFGF], and angiogenin) in the follicular fluid (FF) and oocyte-cumulus complex culture medium (CM) of women undergoing IVF and to investigate the association of the concentrations with the maturity and fertilization of the oocyte. DESIGN: Prospective study. SETTING: Academic tertiary-care institution. PATIENT(S): IVF patients with unexplained or tubal factor infertility. INTERVENTION(S): Analysis of VEGF, bFGF, and angiogenin FF and CM concentrations. MAIN OUTCOME MEASURE(S): Oocyte maturity and fertilization and FF and CM angiogenic factor concentrations. RESULT(S): VEGF, bFGF, and angiogenin were determined in FF and CM. FF angiogenin concentrations were significantly higher when the oocyte was mature versus immature. CM VEGF concentrations were significantly higher when the oocyte was nonfertilized versus fertilized. Positive correlations were observed between angiogenic factors in CM. CONCLUSION(S): VEGF, bFGF, and angiogenin (determined for the first time) are secreted in the FF and CM. Elevated CM VEGF concentrations, probably implying oocyte-cumulus complex hypoxia, are negatively associated with oocyte fertilization. Elevated FF angiogenin concentrations are positively associated with oocyte maturity, possibly indicating angiogenin's biological role beyond neovascularization.

Changes in serum levels of vascular endothelial growth factor in males and females throughout life.

OBJECTIVES: To study serum levels of vascular endothelial growth factor (VEGF), a potent angiogenic factor, during distinct periods of the female life span and compare them with corresponding levels of age-matched males. It is hypothesized that VEGF might be increased at periods of enhanced angiogenesis. METHODS: Venous blood was drawn from healthy females (n = 59) and males (n = 53) divided into six groups: fetuses (cord blood), neonates, children, adults (same females in the proliferative and secretory phases of their menstrual cycle), pregnant, and elderly (postmenopausal). Serum VEGF levels were measured by an enzyme immunoassay. RESULTS: Females showed 49% higher serum VEGF levels than males (t = 2.74, P = .01). Cord and neonatal blood levels were significantly increased compared with those of adults (t = 2.41, P = .02, and t = 5.81, P = .0001, respectively). All female age groups presented higher serum VEGF levels than the group of women in the proliferative phase of the cycle; nevertheless, VEGF levels in the secretory phase did not differ (t = 1.85, P = .07). CONCLUSIONS: Serum VEGF levels are higher in females than in males and during life periods characterized by enhanced growth and development, implying increased rates of angiogenesis.

Adhesion molecules in early neonatal life.

This study investigated whether serum levels of the adhesion molecules VCAM-1, PECAM-1 and L-selectin, all of which have been implicated in normal immune function, change soon after birth. Moreover, the dependence of serum levels of the above-mentioned adhesion molecules on sex and mode of delivery was studied. In healthy neonates, serum levels of L-selectin, PECAM-1, and VCAM-1 do not change during early neonatal life. Thus, significant differences in their serum concentrations soon after birth might imply inflammatory processes or deranged endothelial homeostasis, serving as possible diagnostic markers.

Angiogenic factors in the perinatal period: diversity in biological functions reflected in their serum concentrations soon after birth.

These studies investigated whether serum levels of the angiogenic factors angiogenin, basic fibroblast growth factor (bFGF), and vascular endothelial growth factor (VEGF) change soon after birth due to the elimination of the placenta and to diminished angiogenic but increased adaptational demands in extrauterine life. Also investigated was whether serum levels correlate with sex, birth weight, or mode of delivery. Serum from healthy mothers and their healthy full-term infants at birth (umbilical cord, UC), day 1 (N1) and day 4 (N4) postpartum was analyzed by enzyme immunoassays. Angiogenin levels were higher in maternal serum and rose significantly from UC to N1 and N4, possibly because of the elimination of the placenta, which produces an angiogenin inhibitor. bFGF and VEGF maternal levels were lower than fetal and neonatal ones. Although neonatal bFGF levels did not differ from fetal levels, possibly reflecting diminished angiogenesis ex utero, VEGF levels increased in neonatal serum, possibly signifying adaptational demands. Neither factor depended on sex, mode of delivery, or birth weight. Thus, significant differences from normal reference values of the studied factors might reflect ill-defined situations of the placenta and fetus/newborn serving as early diagnostic markers.

Basic fibroblast growth factor: serum levels in the female.

This study investigated serum levels of basic fibroblast growth factor (b FGF), a potent angiogenic factor, during distinct periods of the female life and compared them with corresponding levels in age-matched males. Healthy females (n = 59) and males (n = 53) were included in the study, divided into six groups: fetuses (cord blood), neonates, children, adults (females in proliferative and secretory phase), pregnant and "elderly" men and women. Serum b FGF levels were measured by an enzyme immunoassay. No statistically significant difference was found between both genders. Blood levels in fetuses and neonates were significantly increased as compared to adults (p = 0.01, p = 0.02, respectively). Restricting the analysis to females, all age groups, but fetuses (p = 0.05), demonstrated no difference when compared to proliferative phase adults. In conclusion, b FGF serum levels do not differ between males and females and are elevated in fetal and neonatal life, when growth and development are enhanced.

Serum angiogenin levels in the female from birth to postmenopause.

This study investigated serum angiogenin levels of the potent angiogenic factor angiogenin, during fetal and neonatal life, childhood, adulthood, pregnancy and postmenopause and compared them with respective levels in age-matched males. Serum angiogenin levels were measured by an enzyme immunoassay in 139 healthy male and female subjects, allocated in the above six groups. Multiple linear regression applied (a) for both genders and (b) only for females showed serum angiogenin levels in adults to differ statistically highly significantly from levels in cord blood (P = 0.0001), neonates (P = 0.0001), children (P = 0.0001), and pregnant women (P = 0.01), but not from "elderly" subjects (P = 0.80). A significant difference existed between levels in the proliferative and secretory phase of the menstrual cycle (P = 0.006). Furthermore, a significant trend for serum angiogenin levels with advancing age groups was noted (P = 0.0001). In conclusion, serum angiogenin levels increase significantly from fetal life to adulthood, possibly implying additional biological functions to that of angiogenesis.

Heparin-binding angiogenic factors (basic fibroblast growth factor and vascular endothelial growth factor) in early neonatal life.

This study investigated whether serum levels of the potent angiogenic factors basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF), which are abundantly produced in utero by the placenta and fetal tissues, change after birth at term, consequent to diminished angiogenic but increased adaptational demands in extrauterine life. Moreover, whether serum levels of the above factors correlate with sex, birth weight, or mode of delivery was also evaluated. One milliliter of blood was drawn from 30 healthy, appropriate for gestational age, full-term infants on d 1 (N1) and 4 (N4) postnatally. In 10 of the above cases maternal and umbilical cord blood samples were also drawn. Serum was analyzed by enzyme immunoassays, using commercial kits. Levels of bFGF and VEGF were significantly lower in maternal serum than in umbilical cord (p = 0.02 and 0.036, respectively) or N1 (p = 0.009 and 0.006, respectively) and N4 serum (p = 0.009 and 0.006, respectively). Levels of bFGF in umbilical cord serum did not differ significantly from those in N1 and N4. In contrast, levels of VEGF rose in N1, differing significantly from levels in umbilical cord serum (p = 0.008). Both factors did not change from N1 to N4. Neither bFGF nor VEGF serum levels depended on sex, mode of delivery, or birth weight. In conclusion, bFGF levels in neonates do not differ from levels in fetuses, possibly reflecting diminished angiogenesis in extrauterine life, which already has started in utero. On the contrary, neonatal levels of VEGF rise significantly after birth, possibly signifying adaptation demands, in addition to angiogenesis, as VEGF is also considered a regulator of normal function.

Serum levels of basic fibroblast growth factor and vascular endothelial growth factor in children and adolescents with type 1 diabetes mellitus.

Diabetes mellitus is characterized by microangiopathy and increased angiogenic response in various organs. Basic fibroblast growth factor (bFGF) as well as vascular endothelial growth factor (VEGF) are both angiogenic and are involved in vascular endothelial cell growth. The purpose of this study was to determine serum levels of bFGF and VEGF, in children and adolescents (youngsters) with type 1 diabetes mellitus, and correlate them with parameters reflecting the severity of the disease. Forty diabetic youngsters without clinical evidence of complications were compared with 30 healthy control subjects (mean age +/- SD, 14.3 +/- 3.6 and 13.8 +/- 3.6 y, respectively). Diabetes duration and metabolic control (expressed by glycosylated Hb) were (mean +/- SD) 6.2 +/- 3.8 y and 9.6 +/- 1.8%, respectively. bFGF and VEGF (pg/mL) were measured in serum samples by enzyme immunoassays, and both were not significantly different between the type 1 diabetes mellitus and the control group (p = 0.952 and p = 0.559, respectively). Restricting the analysis to the type 1 diabetes mellitus group, neither the duration nor the metabolic control of the disease showed any correlation with bFGF and VEGF serum levels, whereas a significantly positive correlation was found between the two examined angiogenic factors both in the diabetic (r = 0.3464, p = 0.025) and the control group (r = 0.4619, p = 0.0013). In conclusion, serum levels of bFGF and VEGF were not found to vary significantly in diabetic youngsters in relation to controls and had no correlation with the duration and metabolic control of the disease. Nevertheless, a positive correlation was found between these two angiogenic factors both in the type 1 diabetes mellitus and the control group.

Serum angiogenin levels in children and adolescents with insulin-dependent diabetes mellitus.

Microangiopathy, one of the most important complications of diabetes mellitus in humans, is associated with increased angiogenic response and proliferative lesions in various organs. Angiogenin, a polypeptide with a molecular size of 14 kD, is a potent inducer of vascular growth. This study aimed at investigating whether serum angiogenin levels are elevated in children and adolescents (youngsters) with insulin-dependent diabetes mellitus and whether angiogenin levels are affected by duration and metabolic control of the disease. It is assumed that angiogenin levels reflect the increased angiogenesis associated with microangiopathy, whether clinically evident or not. Forty diabetic youngsters were compared with 30 healthy control subjects (mean age +/- SD, 14.3 +/- 3.6 y and 13.8 +/- 3.6 y, respectively). The patients' disease duration and glycosylated Hb were (mean +/- SD) 6.2 +/- 3.8 y and 9.6 +/- 1.8%, respectively. Angiogenin (ng/mL) was measured in serum samples by an enzyme immunoassay and was found to be significantly higher (mean +/- SE) in patients (353.3 +/- 20.0) than in control subjects (244.7 +/- 9.6) (p = 0.0002). Levels did not vary with age, but were significantly higher in females compared with male subjects (p = 0.01). In the diabetic youngsters no significant differences were noticed with respect to duration or metabolic control of the disease. In conclusion, serum angiogenin levels were found to be increased among diabetic youngsters, irrespective of the duration and metabolic control of the disease, as well as in female subjects, with or without diabetes.