Fabrizio Proietti - Academia.edu (original) (raw)

Papers by Fabrizio Proietti

Research paper thumbnail of AO-26. Predictive power of oxidative stress markers for the early identification of newborns at high risk for free radicals related diseases

Early Human Development, 2010

P = 0.005], and higher resting beats/min vs. 144 (108-168) beats/min; P = 0.028], compared to con... more P = 0.005], and higher resting beats/min vs. 144 (108-168) beats/min; P = 0.028], compared to controls. Aortic IMT was significantly greater in the IUGR group [0.55 (0.39-0.78) mm vs. 0.36 (0.14-0.56) mm; P < 0.001], after adjustment for birthweight, as well [0.32 (0.17-0.47) mm/kg vs. 0.18 (0.09-0.36) mm/kg; P < 0.001]. IUGR neonates had, also, smaller IAoD [3.7 (2.9-4.4) mm vs. 4.2 (3.3-4.8) mm; P = 0.013], and higher serum BNP levels [218 (26-4870) pg/mL vs. 74 (15-168) pg/mL; P <0.001].

Research paper thumbnail of PP-41. Pain and oxidative stress in the newborn

Early Human Development, 2010

We have tried to determine the brain oxygen saturation normals and to compare cerebral oxymeter a... more We have tried to determine the brain oxygen saturation normals and to compare cerebral oxymeter and pulse oxymeter values in asphyxiated and non-asphyxiated newborns.

Research paper thumbnail of The use of melatonin in hypoxic-ischemic brain damage: an experimental study

Journal of Maternal-Fetal and Neonatal Medicine, 2012

Research paper thumbnail of Resuscitating preterm infants with 100% oxygen is associated with higher oxidative stress than room air

Acta Paediatrica, 2015

The starting fraction of inspired oxygen for preterm resuscitation is a matter of debate and the ... more The starting fraction of inspired oxygen for preterm resuscitation is a matter of debate and the use of room air in full-term asphyxiated infants reduces oxidative stress. This study compared oxidative stress in preterm infants randomised for resuscitation with either 100% oxygen or room air titrated to internationally recommended levels of preductal oxygen saturations.

Research paper thumbnail of Effects of epidural and systemic maternal analgesia in term infants: the NoPiL study

Frontiers in bioscience (Elite edition), 2010

The aim of the No Pain in Labour (NoPiL) study was to evaluate the stress and clinical outcome of... more The aim of the No Pain in Labour (NoPiL) study was to evaluate the stress and clinical outcome of infants vaginally born without maternal analgesia and after maternal epidural or systemic analgesia. We studied 120 healthy term infants, 41 in the no analgesia group, 38 in the epidural analgesia group, and 41 in the systemic analgesia group. Cortisol, beta-endorphin, oxidative stress markers (ie: total hydroperoxide (TH) and advanced oxidation protein products (AOPP)), interleukin-1beta (IL-1beta), and interleukin-8 (IL-8) cytokines were measured in arterial cord blood samples. Infants in the 3 groups had similar Apgar score, cord blood pH and occurrence of hypoglycaemia, hyperbilirubinemia, and respiratory depression. Cortisol and endorphin plasma levels did not differ in the groups, nor did TH and AOPP values. IL-1beta and IL-8 cytokine were higher in infants born after maternal epidural analgesia than in other groups. Short-term outcome and stress were similar in infants vaginally ...

Research paper thumbnail of Antioxidant strategies in genetic syndromes with high neoplastic risk in infant age

Tumori

Oxidative stress plays a key role in carcinogenesis. Oxidative damage to cell components can lead... more Oxidative stress plays a key role in carcinogenesis. Oxidative damage to cell components can lead to the initiation, promotion and progression of cancer. Oxidative stress is also a distinctive sign in several genetic disorders characterized by a cancer predisposition such as ataxia-telangiectasia, Fanconi anemia, Down syndrome, Beckwith-Wiedemann syndrome and Costello syndrome. Taking into account the link between oxidative stress and cancer, the capacity of antioxidant agents to prevent or delay neoplastic development has been tested in various studies, both in vitro and in vivo, with interesting and promising results. In recent years, research has been conducted into the molecular mechanisms linking oxidative stress to the pathogenesis of the genetic syndromes we consider in this review, with the resulting identification of possible new therapeutic targets. The aim of this review is to focus on the oxidative mechanisms intervening in carcinogenesis in cancer-prone genetic disorder...

Research paper thumbnail of A metabolomic study of preterm and term human and formula milk by proton MRS analysis: preliminary results

Journal of Maternal-Fetal and Neonatal Medicine, 2014

Objective: To investigate changes in global metabolic profile between: 1 -breast milk and formula... more Objective: To investigate changes in global metabolic profile between: 1 -breast milk and formula milk, 2 -breast milk from mothers delivering at different gestational age (GA) collected within one week from delivery, and then week by week until term equivalent age. Methods: Proton magnetic resonance spectroscopy (MRS) was used to analyze the watersoluble and lipid fractions extracted from 50 milk samples, 46 human milk at different GA, from 23 weeks of gestation until term equivalent age and four different formula milks. Results: The formula milk for premature infants was the most similar to breast milk of preterm babies. Breast milk showed higher lactose concentrations than formula milk, that conversely presented higher galactose 1-phosphate and maltose concentrations. Mother's milk of very preterm babies (23-25 wks of GA) showed a different metabolic profile from preterm infants !29 wks of GA with a subsequent trend to similarity around the 30th week of post-natal age. Breast milk from preterm infants of 29-34 wks, collected up to 40 wks of post-natal age showed a temporal change over the first three weeks of lactation, approaching to zero with the achievement of term age. Conclusions: Metabolome is a promising tool to study human and artificial milk global metabolic profile.

Research paper thumbnail of Beckwith-Wiedemann syndrome: potassium ascorbate with ribose therapy in a syndrome with high neoplastic risk

Anticancer research, 2011

Beckwith-Wiedemann Syndrome (BWS) is a genomic imprinting disorder characterized by overgrowth an... more Beckwith-Wiedemann Syndrome (BWS) is a genomic imprinting disorder characterized by overgrowth and increased risk of malignancy. We studied the oxidative stress (OS) pattern of our patients with BWS and administered, for the first time, potassium ascorbate with ribose (PAR) once a day as long-term therapy in order to correct the effects induced by free radicals. We describe the clinical features of three patients examined every three months in our clinic. OS was ascertained by measuring a panel of OS biomarkers: non-protein-binding iron, total hydroperoxides, advanced oxidation protein products, isoprostanes, carbonyl groups and thiols. After the presence of OS was established, treatment with PAR was started at the dosage of 300 mg of Potassium Bicarbonate and 150 mg of Ascorbic Acid in aqueous solution and changes occurring in OS biomarkers were followed dosing every three months. Our patients showed higher levels of OS biomarkers than controls at the time of diagnosis. There was a...

Research paper thumbnail of Melatonin reduces endoplasmic reticulum stress and preserves sirtuin 1 expression in neuronal cells of newborn rats after hypoxia-ischemia

Journal of Pineal Research, 2014

Conditions that interfere with the endoplasmic reticulum (ER) functions cause accumulation of unf... more Conditions that interfere with the endoplasmic reticulum (ER) functions cause accumulation of unfolded proteins in the ER lumen, referred to as ER stress, and activate a homeostatic signaling network known as unfolded protein response (UPR). We have previously shown that in neonatal rats subjected to hypoxia-ischemia (HI), melatonin administration significantly reduces brain damage. This study assessed whether attenuation of ER stress is involved in the neuroprotective effect of melatonin after neonatal HI. We found that the UPR was strongly activated after HI. Melatonin significantly reduced the neuron splicing of XBP-1 mRNA, the increased phosphorylation of eIF2a, and elevated expression of chaperone proteins GRP78 and Hsp70 observed after HI in the brain. CHOP, which plays a convergent role in the UPR, was reduced as well. Melatonin also completely prevented the depletion of SIRT-1 induced by HI, and this effect was observed in the same neurons that over-express CHOP. These results demonstrate that melatonin reduces ER stress induced by neonatal HI and preserves SIRT-1 expression, suggesting that SIRT-1, due to its action in the modulation of a wide variety of signaling pathways involved in neuroprotection, may play a key role in the reduction of ER stress and neuroprotection observed after melatonin.

Research paper thumbnail of Development of a fast and simple LC-MS/MS method for measuring the F2-isoprostanes in newborns

Journal of Maternal-Fetal and Neonatal Medicine, 2012

Quantification of F(2)-IsoPs isomer has been regarded as the &amp;amp;amp;amp;amp;amp;amp;amp... more Quantification of F(2)-IsoPs isomer has been regarded as the &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;quot;gold standard&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;quot; to assess oxidative stress status in various adult and neonatal human diseases. These methods require high amounts of plasma. To develop a fast and simple LC-MS/MS method for measuring F(2)-isoprostanes in newborns. A sample of heel blood (0.4 mL) was collected in a tube containing EDTA was collected from 20 term healthy newborns. For measurements, the tandem mass spectrometer has been run in multiple reaction monitoring (MRM) with the electrospray source operating in negative ion mode, and by exploiting the transitions m/z 353.3 &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;gt; 193.2 for F2-IsoPs and 357.3 &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;gt; 197.2 for the internal standard d4-8-iso PGF2α. The concentration of F(2)-IsoPs (in pg/mL) in the collected cord bloods was 60.50 ± 25.04 (mean ± S.D.). No statistical difference was found between male (57.09 ± 19.69) and female (64.67 ± 31.13) concentrations. The overall efficiency of the extraction has been over 80%, while the recovery on spiked samples has been around 94% for spikes of 100 pg/mL with a C.V. of 7.7%. We developed a suitable method for large-scale studies with a reduced sample requirement as it is mandatory in neonatal age. Small samples and quick answers are very useful in Neonatology allowing early diagnosis and preventive treatments&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39; strategies of free radical related diseases of the newborn.

Research paper thumbnail of May oxidative stress biomarkers in cord blood predict the occurrence of necrotizing enterocolitis in preterm infants?

Journal of Maternal-Fetal and Neonatal Medicine, 2012

Oxidative stress (OS) is strongly involved in the pathogenesis of many preterm newborn diseases; ... more Oxidative stress (OS) is strongly involved in the pathogenesis of many preterm newborn diseases; this is due to the low efficiency of neonatal antioxidant systems unable to counteract the harmful effects of free radicals (FRs). Hypoxic-ischemic events and inflammation, involved in necrotizing enterocolitis (NEC) pathogenesis, are responsible of the overproduction of FRs, generating OS. To test the hypotesis that OS markers levels in cord blood may early identify the newborns at high risk to develop NEC. 332 preterm newborns of gestational age (GA) between 24 and 33 week and birth weight (BW) between 460 and 2540 g were consecutively recruited in three european neonatal intensive care units. Markers of potential OS risk: non-protein bound iron (NPBI), and markers of FRs damage: advanced oxidation protein products (AOPP) and total hydroperoxides (TH), were measured in the cord blood. Associations between NEC and OS markers were checked through inferential analysis. Out of 332 preterm babies, 29 developed NEC. Babies with NEC had a BW and a GA significantly lower than healthy babies. AOPP, TH and NPBI cord blood levels were significantly higher in babies with NEC than in babies without (respectively mean AOPP = 28.05 ± 21 vs 15.80 ± 7.14; p &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.05; TH = 154.48 ± 84.67 vs 107.40 ± 61.01; p &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.05; NPBI = 2.21 ± 3.98 vs 0.95 ± 1.59; p &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.05). The determination of OS biomarkers in cord blood can be useful in identifying babies at high risk for NEC and in devising new strategies to ameliorate perinatal outcome.

Research paper thumbnail of Isoprostane levels in urine of preterm newborns treated with ibuprofen for patent ductus arteriosus closure

Pediatric Nephrology, 2011

Patent ductus arteriosus (PDA) is the most common cardiovascular abnormality of the preterm infan... more Patent ductus arteriosus (PDA) is the most common cardiovascular abnormality of the preterm infant usually treated with ibuprofen (IBU). PDA is strictly related to oxidative stress (OS) in neonates. This study tests the hypothesis that OS occurs in neonates with PDA and that IBU treatment reduces OS. Forty-three preterm babies with gestational age (GA) &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt;33 weeks were studied prospectively. Three urine samples were collected: at time 0 (before starting treatment), time 1 (after pharmacological PDA closure), and time 2 (7 days after the end of treatment) in all patients. OS was studied by measuring urinary isoprostane (IPs) levels. The results showed significant changes in urinary IP levels from time 0 to time 2 (Kruskal-Wallis, p=0.047). Time trend showed a significant decrease in IPs from time 0 to time 1 after IBU therapy (p=0.0067). This decrease was followed by an increase in IPs levels 7 days after treatment. IBU therapy for PDA closure reduced the risk of OS related to free-radical (FR) generation. This antioxidant effect of IBU may be beneficial in preterm babies with PDA who are at high risk for OS.

Research paper thumbnail of Inhibition of rapamycin-induced autophagy causes necrotic cell death associated with Bax/Bad mitochondrial translocation

Neuroscience, 2012

Rapamycin, a lipophilic macrolide antibiotic, has been found to reduce injury in different models... more Rapamycin, a lipophilic macrolide antibiotic, has been found to reduce injury in different models of neurodegenerative disorders. We have previously shown that in neonatal rats subjected to hypoxia-ischemia (HI) the neuroprotective effect of rapamycin was associated with increased autophagy and decreased caspase-3 activation. We show here that the strong reduction of caspase-3 activation after rapamycin was due, at least in part, to its effect on the intrinsic apoptotic mitochondrial pathway because after rapamycin treatment there was a marked reduction of Bax and Bad translocation to mitochondria, cytochrome c release, and caspase-3 activation. Poly (ADP-ribose) polymerase 1 (PARP-1) cleavage and the number of terminal dUDP nick-end labeling (TUNEL)-positive cells were also reduced. To assess how the antiapoptotic effect of rapamycin was linked to the strong autophagy signal induced by the drug, we blocked the formation of autophagosomes with 3-methyladenine (3MA). 3MA administered 10 min after rapamycin, elicited again Bax and Bad translocation to the mitochondria but did not cause cytochrome c release and caspase-3 activation. After 3MA treatment, cells underwent necrotic cell death. These data indicate that rapamycin administered before HI prevents the apoptotic signaling taking place through the mitochondrial pathway. We hypothesize that rapamycin confers a preconditioning-like protection and suggest that caution is necessary before using pharmacological agents targeting autophagy in neuroprotection because they could interfere with endogenous protective mechanisms.

Research paper thumbnail of Effects of Lutein on Oxidative Stress in the Term Newborn: A Pilot Study

Research paper thumbnail of Oxidative stress in children affected by epileptic encephalopathies

Journal of the Neurological Sciences, 2011

Oxidative stress may lead to abnormal peroxidation of membrane lipids, oxidation of sulfhydryl gr... more Oxidative stress may lead to abnormal peroxidation of membrane lipids, oxidation of sulfhydryl groups and disruption of nucleic acids. Experimental and clinical studies suggested that free radicals may be involved in the pathogenesis of epilepsy. Three groups of patients were considered in the study. Group 1 (N= 34) included patients affected by epileptic encephalopathy; Group 2 (N= 31) included those affected by idiopathic epilepsy syndromes and under valproic acid (VPA) monotherapy, and Group 3 (N = 22) represented by healthy controls. All patients and healthy children underwent blood withdrawals to evaluate redox status by measuring levels of F2-isoprostanes (F2-iso), advanced oxidative protein products (AOPP), non-protein binding iron (NPBI), thyols (-SH groups), and total hydroperoxydes (TH). In comparison to the controls, Group 1 patients showed significantly higher plasma levels of F2-iso, AOPP, and TH. By contrast, no differences there were in the plasma NPBI concentrations. Again, no statistical differences there were in the plasma levels of the oxidative stress markers between patients from Group 2 and normal subjects. Our study showed that patients with epileptic encephalopathy have increased levels of oxidative stress markers. By contrast, normal redox status was observed in patients with idiopathic epilepsy syndromes under long-term VPA monotherapy.

Research paper thumbnail of Melatonin protects from the long-term consequences of a neonatal hypoxic-ischemic brain injury in rats

Journal of Pineal Research, 2008

Research paper thumbnail of Oxidative damage induced by herbicides is mediated by thiol oxidation and hydroperoxides production

Free Radical Research, 2010

Research paper thumbnail of Resuscitation with supplementary oxygen induces oxidative injury in the cerebral cortex

Free Radical Biology and Medicine, 2012

Isoprostanes, neuroprostanes, isofurans, and neurofurans have all become attractive biomarkers of... more Isoprostanes, neuroprostanes, isofurans, and neurofurans have all become attractive biomarkers of oxidative damage and lipid peroxidation in brain tissue. Asphyxia and subsequent reoxygenation cause a burst of oxygen free radicals. Isoprostanes and isofurans are generated by free radical attacks of esterified arachidonic acid. Neuroprostanes and neurofurans are derived from the peroxidation of docosahexanoic acid, which is abundant in neurons and could therefore more selectively represent oxidative brain injury. Newborn piglets (age 12-36 h) underwent hypoxia until the base excess reached À 20 mmol/L or the mean arterial blood pressure dropped below 15 mm Hg. They were randomly assigned to receive resuscitation with 21, 40, or 100% oxygen for 30 min and then ventilation with air. The levels of isoprostanes, isofurans, neuroprostanes, and neurofurans were determined in brain tissue (ng/g) isolated from the prefrontal cortex using gas chromatography-mass spectrometry (GC/MS) with negative ion chemical ionization (NICI) techniques. A control group underwent the same procedures and observations but was not submitted to hypoxia or hyperoxia. Hypoxia and reoxygenation significantly increased the levels of isoprostanes, isofurans, neuroprostanes, and neurofurans in the cerebral cortex. Nine hours after resuscitation with 100% oxygen for 30 min, there was nearly a 4-fold increase in the levels of isoprostanes and isofurans compared to the control group (P ¼ 0.007 and P ¼ 0.001) and more than a 2-fold increase in neuroprostane levels (P ¼0.002). The levels of neuroprostanes and neurofurans were significantly higher in the piglets that were resuscitated with supplementary oxygen (40 and 100%) compared to the group treated with air (21%). The significance levels of the observed differences in neuroprostanes for the 21% vs 40% comparison and the 21% vs 100% comparison were P o 0.001 and P ¼ 0.001, respectively. For neurofurans, the P values of the 21% vs 40% comparison and the 21% vs 100% comparison were P ¼ 0.036 and P ¼ 0.025, respectively. Supplementary oxygen used for the resuscitation of newborns increases lipid peroxidation in brain cortical neurons, a result that is indicative of oxidative brain damage. These novel findings provide new knowledge regarding the relationships between oxidative brain injury and resuscitation with oxygen.

Research paper thumbnail of Increased autophagy reduces endoplasmic reticulum stress after neonatal hypoxia–ischemia: Role of protein synthesis and autophagic pathways

Experimental Neurology, 2014

The endoplasmic reticulum (ER) stress can result from several pathological conditions that pertur... more The endoplasmic reticulum (ER) stress can result from several pathological conditions that perturb ER homeostasis and is characterized by accumulation of unfolded proteins in the ER lumen. To cope with ER stress, cells activate the unfolded protein response (UPR), a protein quality control mechanism aimed at restoring homeostasis. The present study was undertaken to characterize the UPR after neonatal hypoxia/ischemia (HI) and its crosstalk with autophagy. After HI, there was a significant increase of GRP78 and Hsp70 expression, phosphorylation of eIF2α, Xbp-1 mRNA splicing and CHOP expression, revealing severe ER stress and UPR. Increasing autophagy with rapamycin (Rap) significantly reduced the UPR. Rap did not further increase the eIF2α phosphorylation and p70S6 kinase (p70S6K) inactivation induced by HI. After autophagy activation, however, there was a clear co-localization between monodansylcadaverine (MDC)-positive autophagosome-like structures and the ribosomal protein S6 (RPS6), indicating the presence of ribosomes in autophagosomes (ribophagy). We found that the autophagy inhibitor 3-methyladenine administered after Rap treatment completely reverted the increased phosphorylation of eIF2α and p70S6K inactivation, and blocked the formation of autophagosome-like structures restoring the UPR. These results demonstrate that the UPR is strongly activated after neonatal HI. Over-activation of autophagy significantly reduces this response, highlighting the relevance of the cross-talk between ER and the autophagy machinery in this important pathological condition. Furthermore, the presence of ribosome subunits in autophagosome-like structures suggests that increased ribosome turnover through autophagy (ribophagy) may represent an additional mechanism involved in the neuroprotective effect observed after autophagy overactivation.

Research paper thumbnail of PP-209. Beckwith Wiedemann Syndrome: Administration of potassium ascorbate and ribose in a syndrome with high neoplastic risk and elevated oxidative stress biomarkers

Early Human Development, 2010

after birth with generalized hypotonia. Familial history was unremarkable. Pregnancy was unmarked... more after birth with generalized hypotonia. Familial history was unremarkable. Pregnancy was unmarked but an increased need for assisted delivery after vaginal birth took place. Clinical findings included neonatal hypotonia, altered consciousness (lethargy), dolichocephaly, and bilateral cryptorchidism without any other signs of systemic illness. The newborn had decreased spontaneous movements and arousal, a weak cry, poor suck (need for tube feeding), and poor reflexes. Cranial ultrasound showed a temporal bilateral echogenic lesion. MRI of the head confirmed a diagnosis of extensive thrombosis of cerebral sinuses (CSVT) with a partial occlusion of superior sagittal sinus, torcular, transverse and sigmoid sinuses with no parenchymal brain lesion. A detailed hemostatic screening for hypercoagulable states was normal. Because of persistent and unexplained hypotonia Prader-Willi syndrome (PWS) was suspected. Methylation analysis confirmed PWS. FISH analysis excluded a deletion in 15q11-q13 and maternal uniparental disomy (UPD) for chromosome 15 was confirmed. Treatment consisted of rehydration and systemic anticoagulation with LMWH in a therapeutic anti-X level. The hypotonia gradually improved after the acute phase and the neonate was discharged at 28 days of life. On a follow up examination at 2 months of age, MR venography (MRV) revealed a remarkable reduction of thrombus size.

Research paper thumbnail of AO-26. Predictive power of oxidative stress markers for the early identification of newborns at high risk for free radicals related diseases

Early Human Development, 2010

P = 0.005], and higher resting beats/min vs. 144 (108-168) beats/min; P = 0.028], compared to con... more P = 0.005], and higher resting beats/min vs. 144 (108-168) beats/min; P = 0.028], compared to controls. Aortic IMT was significantly greater in the IUGR group [0.55 (0.39-0.78) mm vs. 0.36 (0.14-0.56) mm; P < 0.001], after adjustment for birthweight, as well [0.32 (0.17-0.47) mm/kg vs. 0.18 (0.09-0.36) mm/kg; P < 0.001]. IUGR neonates had, also, smaller IAoD [3.7 (2.9-4.4) mm vs. 4.2 (3.3-4.8) mm; P = 0.013], and higher serum BNP levels [218 (26-4870) pg/mL vs. 74 (15-168) pg/mL; P <0.001].

Research paper thumbnail of PP-41. Pain and oxidative stress in the newborn

Early Human Development, 2010

We have tried to determine the brain oxygen saturation normals and to compare cerebral oxymeter a... more We have tried to determine the brain oxygen saturation normals and to compare cerebral oxymeter and pulse oxymeter values in asphyxiated and non-asphyxiated newborns.

Research paper thumbnail of The use of melatonin in hypoxic-ischemic brain damage: an experimental study

Journal of Maternal-Fetal and Neonatal Medicine, 2012

Research paper thumbnail of Resuscitating preterm infants with 100% oxygen is associated with higher oxidative stress than room air

Acta Paediatrica, 2015

The starting fraction of inspired oxygen for preterm resuscitation is a matter of debate and the ... more The starting fraction of inspired oxygen for preterm resuscitation is a matter of debate and the use of room air in full-term asphyxiated infants reduces oxidative stress. This study compared oxidative stress in preterm infants randomised for resuscitation with either 100% oxygen or room air titrated to internationally recommended levels of preductal oxygen saturations.

Research paper thumbnail of Effects of epidural and systemic maternal analgesia in term infants: the NoPiL study

Frontiers in bioscience (Elite edition), 2010

The aim of the No Pain in Labour (NoPiL) study was to evaluate the stress and clinical outcome of... more The aim of the No Pain in Labour (NoPiL) study was to evaluate the stress and clinical outcome of infants vaginally born without maternal analgesia and after maternal epidural or systemic analgesia. We studied 120 healthy term infants, 41 in the no analgesia group, 38 in the epidural analgesia group, and 41 in the systemic analgesia group. Cortisol, beta-endorphin, oxidative stress markers (ie: total hydroperoxide (TH) and advanced oxidation protein products (AOPP)), interleukin-1beta (IL-1beta), and interleukin-8 (IL-8) cytokines were measured in arterial cord blood samples. Infants in the 3 groups had similar Apgar score, cord blood pH and occurrence of hypoglycaemia, hyperbilirubinemia, and respiratory depression. Cortisol and endorphin plasma levels did not differ in the groups, nor did TH and AOPP values. IL-1beta and IL-8 cytokine were higher in infants born after maternal epidural analgesia than in other groups. Short-term outcome and stress were similar in infants vaginally ...

Research paper thumbnail of Antioxidant strategies in genetic syndromes with high neoplastic risk in infant age

Tumori

Oxidative stress plays a key role in carcinogenesis. Oxidative damage to cell components can lead... more Oxidative stress plays a key role in carcinogenesis. Oxidative damage to cell components can lead to the initiation, promotion and progression of cancer. Oxidative stress is also a distinctive sign in several genetic disorders characterized by a cancer predisposition such as ataxia-telangiectasia, Fanconi anemia, Down syndrome, Beckwith-Wiedemann syndrome and Costello syndrome. Taking into account the link between oxidative stress and cancer, the capacity of antioxidant agents to prevent or delay neoplastic development has been tested in various studies, both in vitro and in vivo, with interesting and promising results. In recent years, research has been conducted into the molecular mechanisms linking oxidative stress to the pathogenesis of the genetic syndromes we consider in this review, with the resulting identification of possible new therapeutic targets. The aim of this review is to focus on the oxidative mechanisms intervening in carcinogenesis in cancer-prone genetic disorder...

Research paper thumbnail of A metabolomic study of preterm and term human and formula milk by proton MRS analysis: preliminary results

Journal of Maternal-Fetal and Neonatal Medicine, 2014

Objective: To investigate changes in global metabolic profile between: 1 -breast milk and formula... more Objective: To investigate changes in global metabolic profile between: 1 -breast milk and formula milk, 2 -breast milk from mothers delivering at different gestational age (GA) collected within one week from delivery, and then week by week until term equivalent age. Methods: Proton magnetic resonance spectroscopy (MRS) was used to analyze the watersoluble and lipid fractions extracted from 50 milk samples, 46 human milk at different GA, from 23 weeks of gestation until term equivalent age and four different formula milks. Results: The formula milk for premature infants was the most similar to breast milk of preterm babies. Breast milk showed higher lactose concentrations than formula milk, that conversely presented higher galactose 1-phosphate and maltose concentrations. Mother's milk of very preterm babies (23-25 wks of GA) showed a different metabolic profile from preterm infants !29 wks of GA with a subsequent trend to similarity around the 30th week of post-natal age. Breast milk from preterm infants of 29-34 wks, collected up to 40 wks of post-natal age showed a temporal change over the first three weeks of lactation, approaching to zero with the achievement of term age. Conclusions: Metabolome is a promising tool to study human and artificial milk global metabolic profile.

Research paper thumbnail of Beckwith-Wiedemann syndrome: potassium ascorbate with ribose therapy in a syndrome with high neoplastic risk

Anticancer research, 2011

Beckwith-Wiedemann Syndrome (BWS) is a genomic imprinting disorder characterized by overgrowth an... more Beckwith-Wiedemann Syndrome (BWS) is a genomic imprinting disorder characterized by overgrowth and increased risk of malignancy. We studied the oxidative stress (OS) pattern of our patients with BWS and administered, for the first time, potassium ascorbate with ribose (PAR) once a day as long-term therapy in order to correct the effects induced by free radicals. We describe the clinical features of three patients examined every three months in our clinic. OS was ascertained by measuring a panel of OS biomarkers: non-protein-binding iron, total hydroperoxides, advanced oxidation protein products, isoprostanes, carbonyl groups and thiols. After the presence of OS was established, treatment with PAR was started at the dosage of 300 mg of Potassium Bicarbonate and 150 mg of Ascorbic Acid in aqueous solution and changes occurring in OS biomarkers were followed dosing every three months. Our patients showed higher levels of OS biomarkers than controls at the time of diagnosis. There was a...

Research paper thumbnail of Melatonin reduces endoplasmic reticulum stress and preserves sirtuin 1 expression in neuronal cells of newborn rats after hypoxia-ischemia

Journal of Pineal Research, 2014

Conditions that interfere with the endoplasmic reticulum (ER) functions cause accumulation of unf... more Conditions that interfere with the endoplasmic reticulum (ER) functions cause accumulation of unfolded proteins in the ER lumen, referred to as ER stress, and activate a homeostatic signaling network known as unfolded protein response (UPR). We have previously shown that in neonatal rats subjected to hypoxia-ischemia (HI), melatonin administration significantly reduces brain damage. This study assessed whether attenuation of ER stress is involved in the neuroprotective effect of melatonin after neonatal HI. We found that the UPR was strongly activated after HI. Melatonin significantly reduced the neuron splicing of XBP-1 mRNA, the increased phosphorylation of eIF2a, and elevated expression of chaperone proteins GRP78 and Hsp70 observed after HI in the brain. CHOP, which plays a convergent role in the UPR, was reduced as well. Melatonin also completely prevented the depletion of SIRT-1 induced by HI, and this effect was observed in the same neurons that over-express CHOP. These results demonstrate that melatonin reduces ER stress induced by neonatal HI and preserves SIRT-1 expression, suggesting that SIRT-1, due to its action in the modulation of a wide variety of signaling pathways involved in neuroprotection, may play a key role in the reduction of ER stress and neuroprotection observed after melatonin.

Research paper thumbnail of Development of a fast and simple LC-MS/MS method for measuring the F2-isoprostanes in newborns

Journal of Maternal-Fetal and Neonatal Medicine, 2012

Quantification of F(2)-IsoPs isomer has been regarded as the &amp;amp;amp;amp;amp;amp;amp;amp... more Quantification of F(2)-IsoPs isomer has been regarded as the &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;quot;gold standard&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;quot; to assess oxidative stress status in various adult and neonatal human diseases. These methods require high amounts of plasma. To develop a fast and simple LC-MS/MS method for measuring F(2)-isoprostanes in newborns. A sample of heel blood (0.4 mL) was collected in a tube containing EDTA was collected from 20 term healthy newborns. For measurements, the tandem mass spectrometer has been run in multiple reaction monitoring (MRM) with the electrospray source operating in negative ion mode, and by exploiting the transitions m/z 353.3 &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;gt; 193.2 for F2-IsoPs and 357.3 &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;gt; 197.2 for the internal standard d4-8-iso PGF2α. The concentration of F(2)-IsoPs (in pg/mL) in the collected cord bloods was 60.50 ± 25.04 (mean ± S.D.). No statistical difference was found between male (57.09 ± 19.69) and female (64.67 ± 31.13) concentrations. The overall efficiency of the extraction has been over 80%, while the recovery on spiked samples has been around 94% for spikes of 100 pg/mL with a C.V. of 7.7%. We developed a suitable method for large-scale studies with a reduced sample requirement as it is mandatory in neonatal age. Small samples and quick answers are very useful in Neonatology allowing early diagnosis and preventive treatments&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39; strategies of free radical related diseases of the newborn.

Research paper thumbnail of May oxidative stress biomarkers in cord blood predict the occurrence of necrotizing enterocolitis in preterm infants?

Journal of Maternal-Fetal and Neonatal Medicine, 2012

Oxidative stress (OS) is strongly involved in the pathogenesis of many preterm newborn diseases; ... more Oxidative stress (OS) is strongly involved in the pathogenesis of many preterm newborn diseases; this is due to the low efficiency of neonatal antioxidant systems unable to counteract the harmful effects of free radicals (FRs). Hypoxic-ischemic events and inflammation, involved in necrotizing enterocolitis (NEC) pathogenesis, are responsible of the overproduction of FRs, generating OS. To test the hypotesis that OS markers levels in cord blood may early identify the newborns at high risk to develop NEC. 332 preterm newborns of gestational age (GA) between 24 and 33 week and birth weight (BW) between 460 and 2540 g were consecutively recruited in three european neonatal intensive care units. Markers of potential OS risk: non-protein bound iron (NPBI), and markers of FRs damage: advanced oxidation protein products (AOPP) and total hydroperoxides (TH), were measured in the cord blood. Associations between NEC and OS markers were checked through inferential analysis. Out of 332 preterm babies, 29 developed NEC. Babies with NEC had a BW and a GA significantly lower than healthy babies. AOPP, TH and NPBI cord blood levels were significantly higher in babies with NEC than in babies without (respectively mean AOPP = 28.05 ± 21 vs 15.80 ± 7.14; p &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.05; TH = 154.48 ± 84.67 vs 107.40 ± 61.01; p &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.05; NPBI = 2.21 ± 3.98 vs 0.95 ± 1.59; p &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.05). The determination of OS biomarkers in cord blood can be useful in identifying babies at high risk for NEC and in devising new strategies to ameliorate perinatal outcome.

Research paper thumbnail of Isoprostane levels in urine of preterm newborns treated with ibuprofen for patent ductus arteriosus closure

Pediatric Nephrology, 2011

Patent ductus arteriosus (PDA) is the most common cardiovascular abnormality of the preterm infan... more Patent ductus arteriosus (PDA) is the most common cardiovascular abnormality of the preterm infant usually treated with ibuprofen (IBU). PDA is strictly related to oxidative stress (OS) in neonates. This study tests the hypothesis that OS occurs in neonates with PDA and that IBU treatment reduces OS. Forty-three preterm babies with gestational age (GA) &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt;33 weeks were studied prospectively. Three urine samples were collected: at time 0 (before starting treatment), time 1 (after pharmacological PDA closure), and time 2 (7 days after the end of treatment) in all patients. OS was studied by measuring urinary isoprostane (IPs) levels. The results showed significant changes in urinary IP levels from time 0 to time 2 (Kruskal-Wallis, p=0.047). Time trend showed a significant decrease in IPs from time 0 to time 1 after IBU therapy (p=0.0067). This decrease was followed by an increase in IPs levels 7 days after treatment. IBU therapy for PDA closure reduced the risk of OS related to free-radical (FR) generation. This antioxidant effect of IBU may be beneficial in preterm babies with PDA who are at high risk for OS.

Research paper thumbnail of Inhibition of rapamycin-induced autophagy causes necrotic cell death associated with Bax/Bad mitochondrial translocation

Neuroscience, 2012

Rapamycin, a lipophilic macrolide antibiotic, has been found to reduce injury in different models... more Rapamycin, a lipophilic macrolide antibiotic, has been found to reduce injury in different models of neurodegenerative disorders. We have previously shown that in neonatal rats subjected to hypoxia-ischemia (HI) the neuroprotective effect of rapamycin was associated with increased autophagy and decreased caspase-3 activation. We show here that the strong reduction of caspase-3 activation after rapamycin was due, at least in part, to its effect on the intrinsic apoptotic mitochondrial pathway because after rapamycin treatment there was a marked reduction of Bax and Bad translocation to mitochondria, cytochrome c release, and caspase-3 activation. Poly (ADP-ribose) polymerase 1 (PARP-1) cleavage and the number of terminal dUDP nick-end labeling (TUNEL)-positive cells were also reduced. To assess how the antiapoptotic effect of rapamycin was linked to the strong autophagy signal induced by the drug, we blocked the formation of autophagosomes with 3-methyladenine (3MA). 3MA administered 10 min after rapamycin, elicited again Bax and Bad translocation to the mitochondria but did not cause cytochrome c release and caspase-3 activation. After 3MA treatment, cells underwent necrotic cell death. These data indicate that rapamycin administered before HI prevents the apoptotic signaling taking place through the mitochondrial pathway. We hypothesize that rapamycin confers a preconditioning-like protection and suggest that caution is necessary before using pharmacological agents targeting autophagy in neuroprotection because they could interfere with endogenous protective mechanisms.

Research paper thumbnail of Effects of Lutein on Oxidative Stress in the Term Newborn: A Pilot Study

Research paper thumbnail of Oxidative stress in children affected by epileptic encephalopathies

Journal of the Neurological Sciences, 2011

Oxidative stress may lead to abnormal peroxidation of membrane lipids, oxidation of sulfhydryl gr... more Oxidative stress may lead to abnormal peroxidation of membrane lipids, oxidation of sulfhydryl groups and disruption of nucleic acids. Experimental and clinical studies suggested that free radicals may be involved in the pathogenesis of epilepsy. Three groups of patients were considered in the study. Group 1 (N= 34) included patients affected by epileptic encephalopathy; Group 2 (N= 31) included those affected by idiopathic epilepsy syndromes and under valproic acid (VPA) monotherapy, and Group 3 (N = 22) represented by healthy controls. All patients and healthy children underwent blood withdrawals to evaluate redox status by measuring levels of F2-isoprostanes (F2-iso), advanced oxidative protein products (AOPP), non-protein binding iron (NPBI), thyols (-SH groups), and total hydroperoxydes (TH). In comparison to the controls, Group 1 patients showed significantly higher plasma levels of F2-iso, AOPP, and TH. By contrast, no differences there were in the plasma NPBI concentrations. Again, no statistical differences there were in the plasma levels of the oxidative stress markers between patients from Group 2 and normal subjects. Our study showed that patients with epileptic encephalopathy have increased levels of oxidative stress markers. By contrast, normal redox status was observed in patients with idiopathic epilepsy syndromes under long-term VPA monotherapy.

Research paper thumbnail of Melatonin protects from the long-term consequences of a neonatal hypoxic-ischemic brain injury in rats

Journal of Pineal Research, 2008

Research paper thumbnail of Oxidative damage induced by herbicides is mediated by thiol oxidation and hydroperoxides production

Free Radical Research, 2010

Research paper thumbnail of Resuscitation with supplementary oxygen induces oxidative injury in the cerebral cortex

Free Radical Biology and Medicine, 2012

Isoprostanes, neuroprostanes, isofurans, and neurofurans have all become attractive biomarkers of... more Isoprostanes, neuroprostanes, isofurans, and neurofurans have all become attractive biomarkers of oxidative damage and lipid peroxidation in brain tissue. Asphyxia and subsequent reoxygenation cause a burst of oxygen free radicals. Isoprostanes and isofurans are generated by free radical attacks of esterified arachidonic acid. Neuroprostanes and neurofurans are derived from the peroxidation of docosahexanoic acid, which is abundant in neurons and could therefore more selectively represent oxidative brain injury. Newborn piglets (age 12-36 h) underwent hypoxia until the base excess reached À 20 mmol/L or the mean arterial blood pressure dropped below 15 mm Hg. They were randomly assigned to receive resuscitation with 21, 40, or 100% oxygen for 30 min and then ventilation with air. The levels of isoprostanes, isofurans, neuroprostanes, and neurofurans were determined in brain tissue (ng/g) isolated from the prefrontal cortex using gas chromatography-mass spectrometry (GC/MS) with negative ion chemical ionization (NICI) techniques. A control group underwent the same procedures and observations but was not submitted to hypoxia or hyperoxia. Hypoxia and reoxygenation significantly increased the levels of isoprostanes, isofurans, neuroprostanes, and neurofurans in the cerebral cortex. Nine hours after resuscitation with 100% oxygen for 30 min, there was nearly a 4-fold increase in the levels of isoprostanes and isofurans compared to the control group (P ¼ 0.007 and P ¼ 0.001) and more than a 2-fold increase in neuroprostane levels (P ¼0.002). The levels of neuroprostanes and neurofurans were significantly higher in the piglets that were resuscitated with supplementary oxygen (40 and 100%) compared to the group treated with air (21%). The significance levels of the observed differences in neuroprostanes for the 21% vs 40% comparison and the 21% vs 100% comparison were P o 0.001 and P ¼ 0.001, respectively. For neurofurans, the P values of the 21% vs 40% comparison and the 21% vs 100% comparison were P ¼ 0.036 and P ¼ 0.025, respectively. Supplementary oxygen used for the resuscitation of newborns increases lipid peroxidation in brain cortical neurons, a result that is indicative of oxidative brain damage. These novel findings provide new knowledge regarding the relationships between oxidative brain injury and resuscitation with oxygen.

Research paper thumbnail of Increased autophagy reduces endoplasmic reticulum stress after neonatal hypoxia–ischemia: Role of protein synthesis and autophagic pathways

Experimental Neurology, 2014

The endoplasmic reticulum (ER) stress can result from several pathological conditions that pertur... more The endoplasmic reticulum (ER) stress can result from several pathological conditions that perturb ER homeostasis and is characterized by accumulation of unfolded proteins in the ER lumen. To cope with ER stress, cells activate the unfolded protein response (UPR), a protein quality control mechanism aimed at restoring homeostasis. The present study was undertaken to characterize the UPR after neonatal hypoxia/ischemia (HI) and its crosstalk with autophagy. After HI, there was a significant increase of GRP78 and Hsp70 expression, phosphorylation of eIF2α, Xbp-1 mRNA splicing and CHOP expression, revealing severe ER stress and UPR. Increasing autophagy with rapamycin (Rap) significantly reduced the UPR. Rap did not further increase the eIF2α phosphorylation and p70S6 kinase (p70S6K) inactivation induced by HI. After autophagy activation, however, there was a clear co-localization between monodansylcadaverine (MDC)-positive autophagosome-like structures and the ribosomal protein S6 (RPS6), indicating the presence of ribosomes in autophagosomes (ribophagy). We found that the autophagy inhibitor 3-methyladenine administered after Rap treatment completely reverted the increased phosphorylation of eIF2α and p70S6K inactivation, and blocked the formation of autophagosome-like structures restoring the UPR. These results demonstrate that the UPR is strongly activated after neonatal HI. Over-activation of autophagy significantly reduces this response, highlighting the relevance of the cross-talk between ER and the autophagy machinery in this important pathological condition. Furthermore, the presence of ribosome subunits in autophagosome-like structures suggests that increased ribosome turnover through autophagy (ribophagy) may represent an additional mechanism involved in the neuroprotective effect observed after autophagy overactivation.

Research paper thumbnail of PP-209. Beckwith Wiedemann Syndrome: Administration of potassium ascorbate and ribose in a syndrome with high neoplastic risk and elevated oxidative stress biomarkers

Early Human Development, 2010

after birth with generalized hypotonia. Familial history was unremarkable. Pregnancy was unmarked... more after birth with generalized hypotonia. Familial history was unremarkable. Pregnancy was unmarked but an increased need for assisted delivery after vaginal birth took place. Clinical findings included neonatal hypotonia, altered consciousness (lethargy), dolichocephaly, and bilateral cryptorchidism without any other signs of systemic illness. The newborn had decreased spontaneous movements and arousal, a weak cry, poor suck (need for tube feeding), and poor reflexes. Cranial ultrasound showed a temporal bilateral echogenic lesion. MRI of the head confirmed a diagnosis of extensive thrombosis of cerebral sinuses (CSVT) with a partial occlusion of superior sagittal sinus, torcular, transverse and sigmoid sinuses with no parenchymal brain lesion. A detailed hemostatic screening for hypercoagulable states was normal. Because of persistent and unexplained hypotonia Prader-Willi syndrome (PWS) was suspected. Methylation analysis confirmed PWS. FISH analysis excluded a deletion in 15q11-q13 and maternal uniparental disomy (UPD) for chromosome 15 was confirmed. Treatment consisted of rehydration and systemic anticoagulation with LMWH in a therapeutic anti-X level. The hypotonia gradually improved after the acute phase and the neonate was discharged at 28 days of life. On a follow up examination at 2 months of age, MR venography (MRV) revealed a remarkable reduction of thrombus size.