Neurological abnormalities in full-term asphyxiated newborns and salivary S100B testing: the "Cooperative Multitask against Brain Injury of Neonates" (CoMBINe) international study.

BACKGROUND: Perinatal asphyxia (PA) is a leading cause of mortality and morbidity in newborns: its prognosis depends both on the severity of the asphyxia and on the immediate resuscitation to restore oxygen supply and blood circulation. Therefore, we investigated whether measurement of S100B, a consolidated marker of brain injury, in salivary fluid of PA newborns may constitute a useful tool for the early detection of asphyxia-related brain injury. METHODS: We conducted a cross-sectional study in 292 full-term newborns admitted to our NICUs, of whom 48 suffered PA and 244 healthy controls admitted at our NICUs. Saliva S100B levels measurement longitudinally after birth; routine laboratory variables, neurological patterns, cerebral ultrasound and, magnetic resonance imaging were performed. The primary end-point was the presence of neurological abnormalities at 12-months after birth. RESULTS: S100B salivary levels were significantly (P<0.001) higher in newborns with PA than in normal infants. When asphyxiated infants were subdivided according to a good (Group A; n = 15) or poor (Group B; n = 33) neurological outcome at 12-months, S100B was significantly higher at all monitoring time-points in Group B than in Group A or controls (P<0.001, for all). A cut-off >3.25 MoM S100B achieved a sensitivity of 100% (CI5-95%: 89.3%-100%) and a specificity of 100% (CI5-95%: 98.6%-100%) as a single marker for predicting the occurrence of abnormal neurological outcome (area under the ROC curve: 1.000; CI5-95%: 0.987-1.0). CONCLUSIONS: S100B protein measurement in saliva, soon after birth, is a useful tool to identify which asphyxiated infants are at risk of neurological sequelae.

Regulation by hypoxia of adrenomedullin output and expression in human trophoblast cells.

OBJECTIVES: Plasma adrenomedullin concentrations are increased in the fetal circulation in acute and chronic hypoxic conditions. The effect of hypoxia in regulating adrenomedullin synthesis and secretion was investigated in human placental trophoblast cells. STUDY DESIGN: Human trophoblast cells obtained from term placentas (n = 7) were cultured in hypoxic condition (3% oxygen). Cytotrophoblast cells were cultured for up to 48 h and syncytiotrophoblasts for 2, 8 and 24 h. Changes in adrenomedullin output compared to normoxic conditions were measured by radioimmunoassay. Protein expression was evaluated with Western blot and immunocytochemistry. RESULTS: Hypoxia induced a time-dependent increase in adrenomedullin output and protein expression by placental trophoblast cells. CONCLUSIONS: Hypoxia regulates adrenomedullin secretion and expression by human placenta, thereby promoting increased adrenomedullin concentration in the fetal circulation in clinical circumstances characterized by reduced oxygen levels.

Intrafollicular concentration of adrenomedullin is associated with IVF outcome.

OBJECTIVE: To test the hypothesis that serum or intrafollicular concentrations of adrenomedullin (AM) would correlate with reproductive outcomes in in vitro fertilisation (IVF) cycles. DESIGN: Serum and follicular fluid samples were collected during transvaginal oocyte retrieval. The follicular fluid was individually aspirated, and the presence of oocyte was recorded. AM concentrations were measured using an enzyme-linked immunosorbent assay. SETTING: Department of Gynaecology, Perinatology and Child Health, 'Sapienza' University of Rome, Italy. PATIENTS: Eighty women undergoing IVF for primary infertility aged 18-45 years. MAIN OUTCOME MEASURES: AM concentrations in plasma and follicular fluid were correlated to follicular fluid volume, presence of oocyte, oocyte maturation, embryo grading, fertilisation and pregnancy rates, live-birth rate and plasma estrogen concentration. RESULTS: Monofollicular fluid AM concentrations did not differ between follicles containing oocyte and those without oocyte; however, AM concentrations were lower in follicles that resulted in pregnancy than in those that failed. Serum but not follicular fluid AM concentrations correlated with serum estrogen levels. Follicular fluid AM correlated with plasma AM levels. CONCLUSION: We conclude that higher level of AM in the follicular fluid appears to be associated with a negative outcome in IVF treatment.

Effect of corticosteroids and progesterone on adrenomedullin output and expression in human fetal membranes and placenta trophoblasts.

OBJECTIVE: Plasma adrenomedullin (AM) concentrations are increased in fetal and maternal circulation in response to exogenous glucocorticoids administration. The role of corticosteroids and progesterone in regulating AM synthesis and secretion was investigated in amnion and chorion trophoblast cells of the fetal membranes and in placental trophoblast cells. STUDY DESIGN: Cells were treated with betamethasone, hydrocortisone, and progesterone. Changes in AM output were measured with radioimmunoassay. Protein expression was evaluated with Western blot and immunohistochemistry. RESULTS: Betamethasone stimulated AM secretion and protein expression in placental trophoblast cells and in amnion cells of the fetal membranes. Hydrocortisone and progesterone did not induce any effect either on secretion or protein expression in placenta and fetal membranes cells. CONCLUSION: Glucocorticoids regulate AM secretion and expression by human placenta thereby promoting increased AM concentration in maternal and fetal circulation in circumstances characterized by increased cortisol levels.

S100 beta protein is increased in fetuses with neural tube defect.

We investigated the levels of S100 beta protein (S100B) in the serum of fetuses with neural tube defects (NTD), and their mother. Samples from 20 fetuses with NTD and 30 controls at the same gestational age, and their mothers, were studied. S100B protein levels were determined using Lia-mat Sangtec. kit. S100B concentrations were significantly higher in NTD fetuses (median 2.71 microg/L) than in control subjects (median 0.98 microg/L). Increased S100B levels were also found in mothers carrying fetuses with NTD compared to control and uncomplicated pregnancies. This study indicates that NTD is associated with increased serum concentration of S100B in fetuses and mothers. Moreover, it gives information on S100B levels in the fetal circulation in early-mid gestation.

Changes in the interrelationship between leptin, resistin and adiponectin in early neonatal life.

The aim of this study was to investigate the interrelationship between leptin,adiponectin and resistin in the fetal and early postnatal period and the association of these hormones with anthropometric and metabolic indexes. Serum concentrations of leptin, adiponectin and resistin were measured in maternal and neonatal circulation at delivery and on the 3rd day after birth in 40 healthy newborns and their mothers Serum leptin levels were significantly higher in fetuses that in newborn infants on 3rd day after delivery, whereas concentration of adiponectin and resistin were maintained in either maternal and neonatal circulation after delivery. Leptin serum concentrations correlated with those of adiponectin in the fetal circulation, but not in neonatal life. On the other hand no correlation was found between leptin and resistin levels in cord blood, whereas a positive correlation between leptin and resistin concentrations was present in the neonatal circulation on 3rd day. Fetal leptin, adiponectin and resistin levels are largely independent of maternal influences and immediately after birth, important changes in the relation among adipokines occurred compared to intrauterine life.

Biochemical markers of perinatal brain damage.

Hypoxia-ischemia constitutes a risk in infants by altering cerebral blood flow regulatory mechanisms and causing loss of cerebral vascular auto-regulation. Hypotension, cerebral ischemia, and reperfusion are the main events involved in vascular auto-regulation leading to cell death and tissue damage. These dramatic phenomena represent a common repertoire in infants complicated by perinatal acute or chronic hypoxia. To date, despite accurate perinatal and intra-operative monitoring, the post-insult period is crucial, since clinical symptoms and monitoring parameters may be of no avail and therapeutic window for pharmacological intervention (6-12 hours) may be limited, at a time when brain damage is already occurring. Therefore, the measurement of circulating biochemical markers of brain damage, such as vasoactive agents and nervous tissue peptides is eagerly awaited in clinical practice to detect high risk infants. The present review is aimed at investigating the role as circulating biochemical markers such as adrenomedullin, S100B, activin A, neuronal specific enolase (NSE), glial fibrillary acid protein (GFAP), in the cascade of events leading to ischemia reperfusion injury in infants complicated by perinatal asphyxia.

New markers of neonatal neurology.

Hypoxia-ischemia (H-I) constitutes the main phenomenon responsible for brain-blood barrier permeability modifications leading to cerebral vascular auto-regulation loss in newborns. Hypotension, cerebral ischemia, and reperfusion are the main events involved in vascular auto-regulation loss leading to cell death and tissue damage. Reperfusion could be critical since organ damage, particularly of the brain, may be amplified during this period. An exaggerated activation of vasoactive agents, of calcium mediated effects could be responsible for reperfusion injury (R-I), which, in turns, leads to cerebral hemorrhage and damage. These phenomena represent a common repertoire in newborns complicated by perinatal acute or chronic hypoxia treated by risky procedures such as mechanical ventilation, nitric oxide supplementation, brain cooling, and extracorporeal membrane oxygenation (ECMO). Despite accurate monitoring, the post-insult period is crucial, as clinical symptoms and standard monitoring parameters may be silent at a time when brain damage is already occurring and the therapeutic window for pharmacological intervention is limited. Therefore, the measurement of circulating biochemical markers of brain damage, such as vasoactive agents and nervous tissue peptides is eagerly awaited in clinical practice to detect high risk newborns. The present review is aimed at investigating the role of biochemical markers such as adrenomedullin, a vasoactive peptide; S100B, a calcium binding protein, activin A, a glycoprotein, in the cascade of events leading to I-R injury in newborns complicated by perinatal asphyxia.

Circulating biochemical markers of brain damage in infants complicated by ischemia reperfusion injury.

Hypoxia-ischemia constitutes a risk in infants by altering cerebral blood flow regulatory mechanisms and causing loss of cerebral vascular auto-regulation. Hypotension, cerebral ischemia, and reperfusion are the main events involved in vascular auto-regulation leading to cell death and tissue damage. Reperfusion could be critical since organ damage, particularly of the brain, may be amplified during this period. An exaggerated activation of vasoactive agents of calcium mediated effects could be responsible for reperfusion injury, which, in turns, leads to cerebral hemorrhage and damage. These dramatic phenomena represent a common repertoire in infants complicated by perinatal acute or chronic hypoxia or cardiovascular disorders treated by risky procedures such as open heart surgery and cardiopulmonary by-pass (CPB). To date, despite accurate perinatal and intra-operative monitoring, the post-insult period is crucial, since clinical symptoms and monitoring parameters may be of no avail and therapeutic window for pharmacological intervention (6-12 hours) may be limited, at a time when brain damage is already occurring. Therefore, the measurement of circulating biochemical markers of brain damage, such as vasoactive agents and nervous tissue peptides is eagerly awaited in clinical practice to detect high risk infants. The present review is aimed at investigating the role as circulating biochemical markers such as adrenomedullin, a vasoactive peptide; S100B, a calcium binding protein, activin A, a glycoprotein; neuronal specific enolase (NSE), a dimeric isoenzyme; glial fibrillary acid protein (GFAP), a astroglial protein, in the cascade of events leading to ischemia reperfusion injury in infants complicated by perinatal asphyxia or cardiovascular disorders requiring risky therapeutic strategies such as CPB and/or extracorporeal membrane oxygenation.

Effect of calcium ionophore A23187 on prostaglandin synthase type 2 and 15-hydroxy-prostaglandin dehydrogenase expression in human chorion trophoblast cells.

OBJECTIVE: Prostaglandins induce parturition in humans. Prostaglandin output is regulated by the synthetic and metabolic enzymes, prostaglandin synthase type 2 (PTGS2) and 15-hydroxyprostaglandin dehydrogenase (PGDH). The role of calcium in regulating PTGS2 and PGDH expression was investigated in chorion trophoblasts. STUDY DESIGN: Cells were treated with calcium ionophore A23187 in the presence or absence of calcium chelators; changes in messenger ribonucleic acid expression were measured with real-time polymerase chain reaction and analyzed with analysis of variance. Protein expression was evaluated with Western blot and dual immunofluorescence. RESULTS: A23187 stimulated PTGS2 and suppressed PGDH expression. Effects of A23187 were reversed by calcium chelators. PTGS2 had perinuclear and cytosolic distribution, whereas PGDH was cytosolic. Some cells expressed both enzymes, some neither enzyme, and some either PTGS2 or PGDH. CONCLUSION: Chorion cells showed heterogeneity in the expression of PTGS2 and PGDH. Calcium influx regulates PTGS2 and PGDH expression, thereby promoting coordinated increased prostaglandin output in circumstances such as term and preterm labor.

Effects of prenatal betamethasone administration on leptin and adiponectin concentrations in maternal and fetal circulation.

OBJECTIVE: The aim of this study was to determine the effects of in vivo administration of prenatal betamethasone on leptin and adiponectin concentration in maternal and fetal circulation. STUDY DESIGN: Blood samples were collected from 35 pregnant women receiving betamethasone for threatened preterm delivery before and at different time points after drug administration. Cord blood was collected at delivery in infants born from mothers treated with betamethasone and in 15 infants who delivered at the same gestational age not receiving betamethasone. RESULTS: Betamethasone caused an approximately 170% increase in maternal leptin at 24 hours after betamethasone, whereas it had no effects on adiponectin concentration. Betamethasone affects neonatal leptin and adiponectin levels in a time-dependent manner. The glucocorticoid-induced changes in the relationship between these adipokines in maternal and fetal circulation was long lasting. CONCLUSION: These results provide the first evidence for in vivo effects of glucocorticoids on maternal and fetal adipokines relationship in human pregnancy.

Opposite effect of phorbol ester PMA on PTGS2 and PGDH mRNA expression in human chorion trophoblast cells.

Prostaglandins (PGs) induce the mechanism of labor in humans. The enzymes responsible for PG synthesis and metabolism are prostaglandin-endoperoxide synthase 2 (PTGS2) and 15-hydroxyprostaglandin dehydrogenase (PGDH). In human chorion trophoblast cells, calcium ionophore A23187 upregulates PTGS2 and downregulates PGDH protein and mRNA. The authors hypothesize that this regulation requires activation of protein kinase C (PKC) and mitogen-activated protein kinases (MAPKs). Human chorion trophoblasts were incubated with A23187 or phorbol 12-myristate 13-acetate (PMA) in the absence or presence of inhibitors of PKC, c-Jun N-terminal kinase, p38, and MEK1/2. PTGS2 and PGDH mRNA were measured by real-time reverse-transcription polymerase chain reaction. PMA upregulated PTGS2 and downregulated PGDH. The PMA effect was reversed by the inhibition of PKC. The p38 inhibitor reduced the stimulatory effect of PMA and A23187 on PTGS2. MEK1/2 inhibitor reduced the effect of PMA on PTGS2. All MAPK inhibitors failed to reverse the effect of either A23187 or PMA on PGDH. The authors conclude that upon stimulation with the same upstream signals, different downstream intracellular pathways regulate PTGS2 and PGDH mRNA expression.

Adrenomedullin blood concentrations in infants subjected to cardiopulmonary bypass: correlation with monitoring parameters and prediction of poor neurological outcome.

BACKGROUND: Brain injury is a major adverse event after cardiac surgery, especially when extracorporeal circuits are used. We evaluated whether cardiopulmonary bypass (CPB) affects cerebrovascular resistance and plasma concentrations of adrenomedullin (AM), a vasoactive peptide regulating cerebral blood flow. METHODS: We evaluated 50 infants (age <1 year) with congenital heart defects, matched according to a 2-year follow-up; 40 infants had no overt neurological injury, and 10 had brain damage. Blood samples were taken before surgery, during surgery before CPB, at the end of CPB, at the end of surgery, and at 12 h after surgery. Neurological outcome was evaluated before surgery, on postoperative day 7, and 2 years after surgery. We measured AM concentrations and used Doppler velocimetry to measure middle cerebral artery (MCA) pulsatility index (PI). RESULTS: The highest MCA PI values and lowest AM concentrations occurred at the end of CPB and of the surgical procedure. Infants who developed abnormal neurologic sequelae had significantly (P <0.001 for both) higher MCA PI values and lower AM concentrations than patients with normal neurologic outcome at the end of CPB and after surgery. As single markers for predicting neurological abnormalities, AM (cutoff: 17.4 ng/L) achieved a sensitivity of 100% and a specificity of 73.0% and MCA PI (cutoff value: 1.8) a sensitivity of 100% and a specificity of 56.8%. CONCLUSIONS: AM concentrations and MCA PI patterns change during CPB, mainly in infants with brain damage, and may be useful for early identification of infants at risk for brain damage.

Localization and distribution of adrenomedullin receptor in the human placenta: changes with gestational age.

OBJECTIVE: To examine the distribution and localization of adrenomedullin (AM) receptor (AM-R) in human placenta and fetal membranes to assess any change during pregnancy or with labor. STUDY DESIGN: Immunohistochemistry was performed by the avidin/biotin immunoperoxidase method using an antibody specific to AM-R on intrauterine tissues collected from 7-41 weeks of gestation (n=73). RESULTS: AM-R was localized in the placenta and fetal membranes in all 3 trimesters. The distribution of AM-R in the villous and extravillous trophoblast cells of the placenta and in chorion and decidua cells of the fetal membranes changed with gestational age but not with labor. CONCLUSION: AM is secreted by decidua and trophoblast cells that also possess AM-R, suggesting that placental tissues function in both the synthesis and action of AM. Changes in AM-R in the placenta during pregnancy may reflect changes in AM function throughout gestation.

The level of adrenomedullin immunoreactivity in seminal fluid is higher in oligozoospermic subjects and correlates with semen biochemical parameters.

OBJECTIVE: The newly discovered vasoactive peptide, adrenomedullin, and its receptors are widely distributed in various non-vascular tissues. Recent studies have suggested the possible regulatory role of adrenomedullin (AM) at several levels of the pituitary-gonadal axis. We determined the level of adrenomedullin-like immunoreactivity in the seminal fluid and examined its possible correlation with routine semen parameters, semen biochemical levels or plasma levels of FSH, LH, testosterone or prolactin. MATERIALS AND METHODS: A total of 51 males were divided into three groups according to semen analysis: (i) normospermic (n=19); (ii) oligozoospermic (n=17); (iii) azoospermic (n=15). All the subjects were submitted to hormone analysis (LH, FSH, testosterone, prolactin), routine semen parameters and semen biochemical levels (fructosio, citric acid, L-carnitine, nitric oxide) evaluation. AM was determined in plasma and seminal fluid using a specific radioimmunoassay. RESULTS: Mean AM concentration in seminal plasma was higher in oligozoospermic subjects than in normospermic males. In patients with non-obstructive azoospermia AM in semen was significantly lower than in patients with obstructive azoospermia. Semen AM levels correlated negatively with citric acid concentrations in oligozoospermic subjects. In patients with obstructive azoospermia AM in seminal fluid was correlated with citric acid levels. There was a relationship between plasma AM and prolactin. CONCLUSIONS: We conclude that in human seminal fluid AM concentration is increased in infertile oligozoospermic patients and derives very likely from the prostate. Its role in the regulation of male fertility, however has to be understood.

Effect of betamethasone in vivo on placental adrenomedullin in human pregnancy.

OBJECTIVE: The aim of the current study was to determine the effects of in vivo administration of prenatal betamethasone in patients at risk for preterm delivery on adrenomedullin (AM) concentrations in maternal and fetal plasma and on AM localization in placenta and fetal membranes. METHODS: A total of 62 pregnant women between 25 and 35 weeks' gestation were studied. Forty-seven pregnant women received betamethasone (2 x 12 mg intramuscularly given 24 hours apart) for stimulation of fetal lung maturity. Blood samples were collected before betamethasone administration and at different time points after the first and the second dose. Further samples were collected at delivery and, in women who did not deliver, after 1 week and 30 days from betamethasone administration. At delivery, placenta and membranes were collected. Fifteen patients who delivered at the same gestational age not receiving betamethasone represented the control group. AM concentration was determined by radioimmunoassay. Localization of AM in placental tissues was assessed by immunohistochemistry. RESULTS: Betamethasone caused approximately 50% increase in maternal plasma AM at 1 week after administration, whereas in fetal plasma AM levels increased by about 90% at 48 hours after betamethasone administration. There was increased immunohistochemical staining for AM in fetoplacental tissues collected after betamethasone administration. CONCLUSION: These results provide the first evidence for in vivo stimulation of AM, likely of placental origin, by glucocorticoids in the third trimester human pregnancy.

Urinary S100B protein concentrations are increased in intrauterine growth-retarded newborns.

BACKGROUND: Intrauterine growth retardation is one of the major causes of perinatal mortality and morbidity. To date, there are no reliable methods to detect brain damage in these patients. METHODS: We conducted a case-control study in tertiary NICUs from December 2001 to December 2003 with 42 intrauterine growth retardation infants and 84 controls. Routine laboratory variables, neurologic outcome at 7-day follow-up, ultrasound imaging, and urine concentrations of S100B protein were determined at 5 time points. Urine S100B levels were measured by an immunoluminometric assay at first urination, 24, 48, and 72 hours, and 7 days after birth. Routine laboratory parameters and neurologic patterns were assessed at the same time as urine sampling. RESULTS: S100B protein was significantly higher at all of the monitoring time points in urine taken from intrauterine growth retardation newborns than in control infants. When intrauterine growth retardation infants were corrected for the presence of abnormal (group A) or normal (group B) neurologic examination 7 days after birth, S100B was significantly higher at all of the predetermined monitoring time points in group A infants than in group B or controls. At a cutoff of 7.37 multiples of median at first urination, S100B achieved a sensitivity of 95% and a specificity of 99.1% as a single marker for predicting an adverse neurologic outcome. Twenty of 126 patients had neurologic abnormalities, making an overall prevalence of the disease in our population of 15.9% (pretest probability). With respect to the performance of S100B in predicting brain damage, its positive and negative predictive values were 91.0% and 99.0%, respectively. CONCLUSIONS: Increased urine S100B protein levels in intrauterine growth retardation newborns in the first week after birth suggest the presence of brain damage reasonably because of intrauterine hypoxia. Longitudinal S100B protein measurements soon after birth are a useful tool to identify which intrauterine growth retardation infants are at risk of possible neurologic sequelae.

Metastatic pancreatic cancer in late pregnancy: a case report and review of the literature.

The occurrence of pancreatic carcinoma in a young patient is rare and even more so in pregnancy. In this case report, we discuss the presentation and management of pancreatic adenocarcinoma, with lung and liver metastases, diagnosed in a woman in her third trimester of pregnancy (28 weeks). Ultrasound and magnetic resonance imaging scans were carried out and pancreatic mass biopsy during endoscopic retrograde cholangiopancreatography was performed. Severe preeclampsia and fetal growth restriction occurred. A female infant was delivered by cesarean section at 30 weeks of gestation for worsening of maternal clinical conditions and hepatic and pancreatic tests. The patient died 50 days after delivery. Although pancreatic cancer is a very rare event in pregnancy, it should be suspected when epigastric abdominal pain and laboratory parameters suggestive of biliary tract obstruction occur in pregnancy to ensure, at the least, a better pregnancy outcome.

High maternal blood S100B concentrations in pregnancies complicated by intrauterine growth restriction and intraventricular hemorrhage.

BACKGROUND: Intrauterine growth restriction (IUGR) is associated with perinatal mortality and with neurologic damage from intraventricular hemorrhage (IVH). We investigated whether S100B, a neural protein found in high concentrations after cell injury in the nervous system, is increased in serum of women whose pregnancies are complicated by IUGR and whose newborns develop IVH. We also explored the prognostic accuracy of maternal serum S100B for IVH in the newborn. METHODS: We conducted a case-control study of 106 pregnancies complicated by IUGR, including a subgroup (n = 26) who developed IVH after birth, and 212 unaffected pregnancies matched for gestational age. Ultrasound examination, Doppler velocimetry patterns (in the utero-placental vessels and middle cerebral artery), and maternal blood collection were performed before birth; cerebral ultrasound and neurologic examinations were performed after birth. RESULTS: S100B was higher (P <0.001) in IUGR pregnancies complicated by IVH than in those that were not and in controls. At a cutoff of 0.72 microg/L, sensitivity was 100% [95% confidence interval (95% CI), 87%-100%] and specificity was 99.3% (97.5%-99.9%) for prediction of IVH (area under the ROC curve, 0.999). The prevalence of IVH was 8.2% in the whole study population, 93% (95% CI, 83.6%-100%) in those with maternal S100B >0.72 microg/L, and 0% (0%-2.5%) in those with maternal S100B <0.72 microg/L. CONCLUSION: For prediction of IVH, measurements of maternal S100B may be useful at times before clinical, laboratory, and ultrasound patterns can identify risk of IVH.

S100B protein in urine of preterm newborns with ominous outcome.

Prematurity is an important cause of perinatal death, and no reliable biochemical/biophysical markers exist to identify newborns with an increased mortality risk. We aimed to use S100B concentrations in urine as an early indicator of risk of neonatal death. We did a cross-sectional study using urine obtained from 165 preterm newborns, of whom 11 suffered neonatal death within the first week, 121 displayed no overt neurologic syndrome, and 33 suffered neonatal hypoxia and intraventricular hemorrhage (IVH) but not ominous outcome. Urine S100B concentrations were determined at four time-points and corrected for gestational age by conversion to multiples of median (MoM) of healthy controls of the same gestational age. Ultrasound imaging was assessed within the first 72 h from birth. In infants that died within the first week, S100B levels in urine were already higher than controls at first urination and increased progressively between the 24 and 96-h time-points. Multiple logistic regression analysis showed a significant correlation between urine S100B protein concentrations and the occurrence of neonatal death. An S100B concentration cut-off of 12.93 MoM at first urination had a sensitivity of 100% and a specificity of 97.8% for predicting an ominous outcome. The positive predictive value was 78.6%, the negative predictive value was 100%. Measurement of urine S100B protein levels in preterm newborns could be useful to identify newborns at higher risk of neonatal death.