Ettore Capoluongo - Academia.edu (original) (raw)

Papers by Ettore Capoluongo

Investigative Ophthalmology & Visual Science, 2010

To evaluate the functional effect of short-term supplementation of saffron, a spice containing th... more To evaluate the functional effect of short-term supplementation of saffron, a spice containing the antioxidant carotenoids crocin and crocetin, in early age-related macular degeneration (AMD). METHODS. Twenty-five patients with AMD were randomly assigned to oral saffron 20 mg/d or placebo supplementation over a 3-month period and then reverted to placebo or saffron for a further 3 months. Focal electroretinograms (fERGs) and clinical findings were recorded at baseline and after 3 months of saffron or placebo supplementation. fERGs were recorded in response to a sinusoidally modulated (41 Hz), uniform field presented to the macular region (18°) at different modulations between 16.5% and 93.5%. Main outcome measures were fERG amplitude (in microvolts), phase (in degrees), and modulation thresholds. RESULTS. After saffron, patients' fERGs were increased in amplitude, compared with either baseline or values found after placebo supplementation (mean change after saffron, 0.25 log V; mean change after placebo, Ϫ0.003 log V; P Ͻ 0.01). fERG thresholds were decreased after saffron supplementation but not placebo, compared with baseline (mean change after saffron, Ϫ0.26 log units; mean change after placebo, 0.0003 log units).

Case Reports in Genetics, 2015

Cystic fibrosis (CF; OMIM number 219700) is an autosomal recessive disease caused by mutations in... more Cystic fibrosis (CF; OMIM number 219700) is an autosomal recessive disease caused by mutations in the CFTR (cystic fibrosis transmembrane conductance regulator) gene, which results in abnormal viscous mucoid secretions in multiple organs and whose main clinical features are pancreatic insufficiency, chronic endobronchial infection, and male infertility. We report the case of a 47-year-old apparently normal male resulting in homozygosity for the mutation p.M348K from nonconsanguineous parents. The proband was screened using a standard panel of 70 different tested on NanoChip 400 platform. The massive parallel pyrosequencing on 454 JS machine allowed the second level analysis. The patient was firstly screened with two different platforms available in our laboratory, obtaining an ambiguous signal for the p.R347P mutation. For this reason we decided to clarify the discordant result of CFTR status by Next Generation Sequencing (NGS) using 454 Junior instrument. The patient is resulted no carrier of the p.R347P mutation, but NGS highlighted a homozygous substitution from T>A at position 1043 in the coding region, causing an amino acid substitution from methionine to lysine (p.M348K). Casual finding of p.M348K homozygote mutation in an individual, without any feature of classical or nonclassical CF form, allowed us to confirm that p.M348K is a benign rare polymorphism.

Journal of laboratory automation, Jan 13, 2015

Bronchoalveolar lavage (BAL) partially recovers both the instilled saline and the alveolar fluid,... more Bronchoalveolar lavage (BAL) partially recovers both the instilled saline and the alveolar fluid, so-called epithelial lining fluid (ELF), but a correction for the dilution due to the BAL technique itself is needed to know the amount of recovered ELF. In this regard, urea nitrogen may be useful and has been proposed to calculate ELF. The aim of the present study was to develop and validate a new method to measure urea nitrogen in BAL fluid (BALF). We used 19 BALF samples obtained from neonates and infants with different respiratory conditions. The urea nitrogen assay was carried out on Cobas c311 analyzer (Roche Diagnostics). A validation study shows that the method is perfectly linear (R(2) = 0.999), sensitive (limit of detection = 0.055 mg/dL; limit of quantification = 0.16 mg/dL), repeatable (low = 0.15 ± 0.02, 13.3%; high = 1.80 ± 0.02, 1.1%), reproducible (low = 0.14 ± 0.02, 14.2 %; high = 1.76 ± 0.04, 2.2 %) with accuracy ranging between 93-96%. Our results support the robustn...

The European respiratory journal, 2014

Hypothermia has become an evidence-based treatment for neonates with hypoxic-ischaemic encephalop... more Hypothermia has become an evidence-based treatment for neonates with hypoxic-ischaemic encephalopathy (HIE) . The usefulness of hypothermia is due to several mechanisms and among these the reduction in inflammation seems to play a relevant role . When applied as whole body hypothermia (WBH), cooling may affect other organs as well. Recent data showed better respiratory outcomes and trends towards lower inflammation in WBH-treated preterm lambs [4], suggesting its possible usefulness to reduce lung injury through the modulation of the inflammatory pathway. By contrast, experiments with hibernating animals have shown that temperature induces significant adaptive changes to the surfactant composition and structure . Nevertheless, no data are currently available in humans. Two case reports have recently described an infant [6] and an adult [7] with severe lung injury, whose ventilation had been facilitated by concurrent hypothermia. Since hypothermia is an accepted therapy only for HIE, we designed a preliminary translational study to investigate the effect of WBH on inflammation and surfactant status in neonates with HIE unaffected by any pulmonary injury. LETTER IN PRESS | CORRECTED PROOF 1 .

Pediatric Blood & Cancer, 2009

Pediatric Research, 2005

Objective: Immun response against pertussis can be induced by infection and/or vaccination and va... more Objective: Immun response against pertussis can be induced by infection and/or vaccination and vaccine induced immunity is known to wane within the following decade. Our aim was to assess the pertussis immun response among adolescent girls in Edirne province in Turkey and to determine its relationship with some parameters.

Journal of Hepatology, 2011

Thyroid, 2011

Resistance to thyroid hormone (RTH) is a genetic disease characterized by a reduced responsivenes... more Resistance to thyroid hormone (RTH) is a genetic disease characterized by a reduced responsiveness of the pituitary and peripheral target tissues to TH. We describe two patients with RTH in whom differentiated thyroid cancer was diagnosed. In both patients, RTH was unequivocally diagnosed and both underwent thyroidectomy for multinodular goiter. In Patient 1, histology showed a papillary thyroid carcinoma pT2. Because serum thyrotropin (TSH) levels were elevated even while the patient was taking 150 μg daily of levothyroxine (LT(4)), the patient was treated with (131)I 100 mCi for ablation of the thyroid remnant without discontinuing his LT(4) therapy. We obtained a clinically adequate response by administering LT(4) 175 μg/day (2.18 μg/kg), but the serum TSH was persistently elevated on this dose. The patient was considered "free of disease" after 8 years of follow-up. In Patient 2, histology revealed a papillary microcarcinoma (0.6 cm). Diagnostic whole-body scan was performed while the patient was taking 100 μg/day LT(4), a time that his serum TSH was 38 μU/mL. Only a small remnant was revealed, so (131)I remnant ablation was not performed. While taking LT(4) at a dose of 175 μg/day (3 μg/kg), the serum TSH was persistently high, serum TH levels were in the normal-high range, and he appeared to be clinically euthyroid. There has been no evidence of persistent or recurrent thyroid carcinoma in ultrasonography and thyroglobulin measurements that have been performed on a yearly basis for 3 years. Patients with thyroid carcinoma and RTH are a unique model of thyroid cancer in which follow-up likely occurs in the setting of constantly elevated serum TSH concentrations. The concern in these patients is that their persistent elevation of serum TSH may have an adverse effect on their thyroid cancer, and management choices in terms of the dose of LT(4) that provides the optimum lowering of serum TSH without toxicity are difficult, particularly in the situation wherein, as was the case with one of our patients, there is cardiac disease.

Thyroid, 2013

Postpartum thyroiditis (PPT) is characterized by the development of postpartum thyroid dysfunctio... more Postpartum thyroiditis (PPT) is characterized by the development of postpartum thyroid dysfunction, which may occur up to 12 months after delivery. The syndrome usually presents with transient thyrotoxicosis, followed by transient hypothyroidism. The association of this condition with resistance to thyroid hormones (RTH) has never been described. In this report, we describe a 30-year-old patient affected by RTH due to a novel p.V283A thyroid hormone receptor-β (THRB) heterozygous mutation in exon 8, which affects the ligand-binding domain, never before described in literature. A simple polymorphism was excluded through screening of 100 healthy controls. The patient became pregnant twice (in 2008 and in 2009) and developed PPT after both deliveries. Two months after her first pregnancy and one month after her second pregnancy, she presented with severe endogenous thyrotoxicosis and concomitant suppressed thyrotropin (TSH) levels, which represents an unusual finding in patients affected by RTH. Other causes of hyperthyroidism were excluded. After the hyperthyroid phase, she became hypothyroid (TSH >75 mU/L and low free-thyroxine and free-tri-iodothyronine levels), and eventually returned to her usual euthyroid status. During the course of PPT, no specific treatment was required, except for β-blockers used to treat tachycardia during the hyperthyroid phase. We report a unique case of a woman affected by RTH, due to a novel mutation V283A in THRB, who experienced PPT with a severe thyrotoxic phase after both her pregnancies. The association between RTH and PPT has never been reported in the literature. In particular, the marked suppression of TSH occurring when levels of TH are particularly elevated is not a frequent condition during RTH.

Rheumatology International, 2008

The aberrant induction of salivary/lacrimal proteins is considered to be crucial in the pathogene... more The aberrant induction of salivary/lacrimal proteins is considered to be crucial in the pathogenesis of sicca-symptoms related to primary Sjögren syndrome (SS). We report the case of an 11-year-old boy who was admitted to hospital due to recurrent bilateral parotid gland enlargement and keratoconjunctivitis, which were diagnosed as primary SS upon a combination of laboratory and instrumental tests. The proteomic analysis of the salivary peptide complex in the patient's salivary fluid near diagnosis and after 6 months of pharmacological therapy revealed quantitative and mostly qualitative differences. This observation reveals that clinical and functional changes of the salivary glands driven by non-steroidal antinflammatory drugs might be reflected in different proteomic patterns of the salivary fluid.

Platelets, 2005

Plasma homocysteine levels depend in part on the molecular nature of the methylenetetrahydrofolat... more Plasma homocysteine levels depend in part on the molecular nature of the methylenetetrahydrofolate reductase (MTHFR) and on blood folate intake. Little has been reported on platelet counts in the presence of hyperhomocysteinemia and MTHFR polymorphisms, with the exception of delayed platelet recovery in homozygous MTHFR C677T subjects after treatment with methotrexate for ovarian cancer. The aim of this investigation was to evaluate the possibility of a link between the platelet count and plasma homocysteine levels in different MTHFR variants in 165 female patients. Determinations of plasma homocysteine levels were by ELISA and of MTHFR polymorphisms (A1298C and C677T) were by inverse hybridization. Serum P- and E-selectin concentrations were obtained by ELISA. An inverse correlation (R=-0.88, P<0.001) was observed between blood platelet counts and plasma homocysteine levels in the women homozygous for MTHFR C677T. This correlation did not depend on pregnancy or other variables reported. Serum concentrations of sE- and sP-selectin, markers of endothelial and platelet activation, were significantly and positively correlated with homocysteine levels. These findings suggest that homocysteine affects platelet numbers in women with MTHFR C677T possibly consequent to endothelial and platelet activation.

Pediatric Pulmonology, 2006

ABSTRACT

Pediatric Critical Care Medicine, 2011

To verify if secretory phospholipase A2 (sPLA2) is increased in pediatric acute respiratory distr... more To verify if secretory phospholipase A2 (sPLA2) is increased in pediatric acute respiratory distress syndrome (ARDS) triggered or not by respiratory syncytial virus infection and to clarify how the enzyme may influence the disease severity and the degree of ventilatory support. Prospective pilot study. Two academic pediatric intensive care units. All infants < 6 months old hospitalized for severe respiratory syncytial virus bronchiolitis, who developed ARDS (respiratory syncytial virus-ARDS group); all infants < 6 months old diagnosed with ARDS secondary to other causes (ARDS group); and infants < 6 months old who needed ventilation for reasons other than any lung disease (control group). None. We enrolled six respiratory syncytial virus -ARDS babies, five ARDS babies, and six control infants. The sPLA2 activity and tumor necrosis factor (TNF)-α were significantly higher in the bronchoalveolar lavage of ARDS infants. Worst oxygenation, ventilation, and longer pediatric intensive care unit stay and ventilation time were present in ARDS babies. No differences were found in Clara cell secretory protein and in serum cytokines levels. Because there is no correlation between bronchoalveolar lavage protein content (a marker of permeability) and sPLA2, the enzyme seems mainly produced in the alveoli. TNF-α, the main inductor of sPLA2 expression, significantly correlates with the enzyme level in the bronchoalveolar lavage. Significant positive correlations exist between sPLA2, TNF-α and oxygen need, mean airway pressure, ventilatory index, and the Murray's lung injury score. Negative correlations were also found between sPLA2, TNF-α, and Pao2/Fio2 ratio. The sPLA2 and TNF-α are increased in ARDS and seem correlated with clinical severity, higher oxygen requirement, and more aggressive ventilation. This correlation confirms findings from adult experience and should guide further investigations on pediatric ARDS pathophysiology.

Pediatric Anesthesia, 2011

Journal of Separation Science, 2009

Listeria monocytogenes is frequently found as a contaminant in raw and ready-to-eat foods. The ab... more Listeria monocytogenes is frequently found as a contaminant in raw and ready-to-eat foods. The ability of L. monocytogenes to multiply at refrigeration temperatures and to grow in a wide range of pH values is of particular concern for food safety. According to the European Union regulation on microbiological criteria for foodstuffs, L. monocytogenes must be absent in some categories of ready-to-eat foods. The standard microbiological method for L. monocytogenes detection in foods (ISO 11290-1: 1996 (ISO, International Organization for Standardization)) is cost and time consuming. Developments of rapid, cost-effective and automated diagnostic methods to detect food-borne pathogens in foods continue to be a major concern for the industry and public health. The aim of this study was the development of a rapid, sensitive and specific molecular detection method for L. monocytogenes. To this purpose, we have applied a capillary electrophoresis method to a PCR protocol (PCR-EES (EES, experion automated electrophoresis system)) for detecting L. monocytogenes in food. In particular, a microfluidic chip-based automated electrophoresis system (experion automated electrophoresis system, Bio-Rad Laboratories, USA) was used for the rapid and automatic analysis of the amplicons. Fifty naturally contaminated samples were analysed with this method and the results were compared with those obtained with ISO method. Moreover, the microfluidic chip-based automated electrophoresis system was compared with classical gel electrophoresis (PCR-CGE). The results showed that after 24 h of culture enrichment, the PCR-EES showed a relative accuracy of 100% with ISO, while using PCR-CGE decreased it down to 96%. After 48 h of enrichment, both PCR-EES and PCR-CGE showed an accuracy of 100% with ISO.

Journal of Separation Science, 2008

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is one of the most wellknown human genetic de... more Glucose-6-phosphate dehydrogenase (G6PD) deficiency is one of the most wellknown human genetic defects, identified in more than 400 million individuals in the world. To date, no commercial kits are available for the mutation screening of this disease. Seventy G6PD-deficient Italian individuals admitted to the Laboratory of Clinical Molecular Biology of Hospital "Agostino Gemelli" of Rome were screened for the most frequent Italian mutations, by means of allele-specific PCR, followed by restriction fragment length electrophoresis. The present study compares two techniques for the identification of restriction patterns: agarose gel electrophoresis versus Experion system. When the first screening was negative, the entire G6PD gene was sequenced using the ABI 3100 Avant Instrumentation. The G6PD variants identified and their frequencies were the following: G6PD Mediterranean (75.7%), G6PD Seattle (7.1%), G6PD A -202 + 376 (7.1%), and G6PD Cassano (2.8%). In addition, we identified by direct sequencing two new mutations, namely Buenos Aires and Rignano. With the Experion method, the size band determination was more accurate than that obtained by gel electrophoresis. The Experion system resulted as a valid, easy, and reproducible diagnostic method for the screening of G6PD mutation as compared with the agarose electrophoretic analysis.

Journal of Proteome Research, 2007

A 1 year follow-up investigation of salivary acidic proline-rich proteins (aPRPs) in preterm and ... more A 1 year follow-up investigation of salivary acidic proline-rich proteins (aPRPs) in preterm and at-term newborns using HPLC-ESI-IT-MS showed that (i) this class of proteins is constitutive rather than inducible, as it is still found in the oral cavity of preterm newborns from 180 days of postconception age (PCA); (ii) the expression of PRH-2 locus anticipates that of PRH-1, since Db isoforms are expressed some months after the PRP-1 and PRP-2 isoforms. The evaluation of the relative abundances of the different aPRPs isoforms and derivatives (differently phosphorylated and cleaved) as a function of PCA showed that (iii) the proteolytic enzymes generating truncated isoforms are also constitutive because they are fully active since 180 days of PCA; (iv) the kinase involved in aPRP phosphorylation is not fully mature in preterm newborns, but its activity increases with PCA, synchronizing with that of at-term newborns and reaching the adult levels at about 500-600 days of PCA, in concomitance with the beginning of deciduous dentition.

IUBMB Life, 2009

Steroid 21-hydroxylase deficiency is present in more than 90% of patients with congenital adrenal... more Steroid 21-hydroxylase deficiency is present in more than 90% of patients with congenital adrenal hyperplasia (CAH), an inherited metabolic disorder of adrenal steroidogenesis. Impaired enzymatic activity leads to the accumulation of metabolic intermediates (progesterone and 17-hydroxyprogesterone), which results in excessive androgen production and varied signs of virilisation. CYP21A2 is an active gene and encodes for the steroid 21-hydroxylase enzyme, whereas CYP21A1P is an inactive pseudogene that contains a series of deleterious mutations. The major part of disease-causing mutations in CYP21A2 alleles are CYP21A1P-derived sequence transferred to the active gene by macro or microconversion events. Only around 5% of all disease-causing CYP21A2 alleles harbour rare mutations that do not originate from the pseudogene. In this report, we report the characterisation of two novel CYP21A2 missense mutations (p.H119R and p.I194N) found in two unrelated Italian patients with nonclassic (NC) CAH clinical diagnosis. Functional in vitro assays for mutagenized CYP21 enzymes were performed in transiently transfected mammalian cells to test the residual enzyme activity and the apparent kinetic values. The residual activities obtained for p.H119R and p.I194N mutants allowed to classify them as NC-CAH associated mutations. These results correlate with the rate of severity of the patients' disease. Finally, the new p.H119R and p.I194N mutations should be included in the panel of those already listed for association with the NC form of 21-hydroxylase deficiency. 2009 IUBMB IUBMB Life, 61(3): 229-235, 2009

Intensive Care Medicine, 2007

Mannose-binding lectin (MBL2) is a collectin molecule able to activate the complement system and ... more Mannose-binding lectin (MBL2) is a collectin molecule able to activate the complement system and the subsequent inflammatory mechanisms. Several MBL2 genetic variants have been described, including the six variants studied in this report, which are those analyzed in most detail in the medical literature. The present study analyzes the prevalence of MBL2 gene variants in preterm newborns and associates individual genotypes with pulmonary outcome variables. All polymorphisms were analyzed by means of a commercially available reverse dot-blot kit. Tertiary neonatal intensive care unit. Seventy-five consecutive preterm newborns. Two variants were particularly analyzed: -550G > C and R52C. The first one is known to be associated with lower protein synthesis when included in specific haplotypes. The homozygous and heterozygous -550G > C mutations were significantly associated with protective effects regarding different lung outcome variables, including shorter duration of mechanical ventilation, hours of continuous positive airway pressure and lower number of hemotransfusions. In contrast, the heterozygous R52C mutation was associated with unfavorable outcome, including higher bronchopulmonary dysplasia prevalence. Multivariate logistic regression analysis showed that these associations were independent of gestational age and birth weight. In addition, four groups of patients were defined on the basis of haplotype combinations. Those known to be associated with low serum MBL2 levels were linked to a better outcome in terms of factors such as hours of mechanical ventilation, continuous positive airway pressure, number of hemotransfusions and bronchopulmonary disease development. The four haplotype combination groups may have a potential diagnostic use as opposite risk factors for lung disease of prematurity.

Intensive Care Medicine, 2006

In addition to the previous classification of chronic lung disease (CLD) O2 dependency at 36 week... more In addition to the previous classification of chronic lung disease (CLD) O2 dependency at 36 weeks of postmenstrual age, a new definition of CLD has recently been proposed: new bronchopulmonary-dysplasia (BPD). This uses total duration of O2 supplementation and positive pressure requirements to delineate three degrees of severity (mild, moderate, and severe) according to the respiratory status at 36 weeks postmenstrual age. We analyzed the balance of serum proinflammatory and profibrotic/angiogenic cytokine concentrations in relation to CLD and the new BPD definition. Descriptive study in a third-level neonatal ICU. Thirty-one preterm neonates with a gestational age of 24-29 weeks were studied to evaluate their serum cytokine concentration; they were previously enrolled in a randomized clinical trial to compare the effects of high-frequency oscillatory ventilation vs. intermittent mandatory ventilation in terms of pulmonary mechanics and lung cytokines. Serum samples were collected on days 1, 3, and 5 after birth until extubation to detect the levels of three proinflammatory cytokines plus four profibrotic/angiogenic cytokines, and correlations were examined to old CLD and new BPD. Ventilation treatments were distributed homogeneously between the groups and did not interfere with the results presented here. Old CLD development, mainly corresponding to the moderate/severe forms of new BPD, was associated with increased proinflammatory and profibrotic/angiogenic cytokines, while mild forms of new BPD were characterized only by increases in profibrotic/angiogenic cytokines, suggesting a different balance of two pathogenic mechanisms in different phases of the disease.

Investigative Ophthalmology & Visual Science, 2010

To evaluate the functional effect of short-term supplementation of saffron, a spice containing th... more To evaluate the functional effect of short-term supplementation of saffron, a spice containing the antioxidant carotenoids crocin and crocetin, in early age-related macular degeneration (AMD). METHODS. Twenty-five patients with AMD were randomly assigned to oral saffron 20 mg/d or placebo supplementation over a 3-month period and then reverted to placebo or saffron for a further 3 months. Focal electroretinograms (fERGs) and clinical findings were recorded at baseline and after 3 months of saffron or placebo supplementation. fERGs were recorded in response to a sinusoidally modulated (41 Hz), uniform field presented to the macular region (18°) at different modulations between 16.5% and 93.5%. Main outcome measures were fERG amplitude (in microvolts), phase (in degrees), and modulation thresholds. RESULTS. After saffron, patients' fERGs were increased in amplitude, compared with either baseline or values found after placebo supplementation (mean change after saffron, 0.25 log V; mean change after placebo, Ϫ0.003 log V; P Ͻ 0.01). fERG thresholds were decreased after saffron supplementation but not placebo, compared with baseline (mean change after saffron, Ϫ0.26 log units; mean change after placebo, 0.0003 log units).

Case Reports in Genetics, 2015

Cystic fibrosis (CF; OMIM number 219700) is an autosomal recessive disease caused by mutations in... more Cystic fibrosis (CF; OMIM number 219700) is an autosomal recessive disease caused by mutations in the CFTR (cystic fibrosis transmembrane conductance regulator) gene, which results in abnormal viscous mucoid secretions in multiple organs and whose main clinical features are pancreatic insufficiency, chronic endobronchial infection, and male infertility. We report the case of a 47-year-old apparently normal male resulting in homozygosity for the mutation p.M348K from nonconsanguineous parents. The proband was screened using a standard panel of 70 different tested on NanoChip 400 platform. The massive parallel pyrosequencing on 454 JS machine allowed the second level analysis. The patient was firstly screened with two different platforms available in our laboratory, obtaining an ambiguous signal for the p.R347P mutation. For this reason we decided to clarify the discordant result of CFTR status by Next Generation Sequencing (NGS) using 454 Junior instrument. The patient is resulted no carrier of the p.R347P mutation, but NGS highlighted a homozygous substitution from T>A at position 1043 in the coding region, causing an amino acid substitution from methionine to lysine (p.M348K). Casual finding of p.M348K homozygote mutation in an individual, without any feature of classical or nonclassical CF form, allowed us to confirm that p.M348K is a benign rare polymorphism.

Journal of laboratory automation, Jan 13, 2015

Bronchoalveolar lavage (BAL) partially recovers both the instilled saline and the alveolar fluid,... more Bronchoalveolar lavage (BAL) partially recovers both the instilled saline and the alveolar fluid, so-called epithelial lining fluid (ELF), but a correction for the dilution due to the BAL technique itself is needed to know the amount of recovered ELF. In this regard, urea nitrogen may be useful and has been proposed to calculate ELF. The aim of the present study was to develop and validate a new method to measure urea nitrogen in BAL fluid (BALF). We used 19 BALF samples obtained from neonates and infants with different respiratory conditions. The urea nitrogen assay was carried out on Cobas c311 analyzer (Roche Diagnostics). A validation study shows that the method is perfectly linear (R(2) = 0.999), sensitive (limit of detection = 0.055 mg/dL; limit of quantification = 0.16 mg/dL), repeatable (low = 0.15 ± 0.02, 13.3%; high = 1.80 ± 0.02, 1.1%), reproducible (low = 0.14 ± 0.02, 14.2 %; high = 1.76 ± 0.04, 2.2 %) with accuracy ranging between 93-96%. Our results support the robustn...

The European respiratory journal, 2014

Hypothermia has become an evidence-based treatment for neonates with hypoxic-ischaemic encephalop... more Hypothermia has become an evidence-based treatment for neonates with hypoxic-ischaemic encephalopathy (HIE) . The usefulness of hypothermia is due to several mechanisms and among these the reduction in inflammation seems to play a relevant role . When applied as whole body hypothermia (WBH), cooling may affect other organs as well. Recent data showed better respiratory outcomes and trends towards lower inflammation in WBH-treated preterm lambs [4], suggesting its possible usefulness to reduce lung injury through the modulation of the inflammatory pathway. By contrast, experiments with hibernating animals have shown that temperature induces significant adaptive changes to the surfactant composition and structure . Nevertheless, no data are currently available in humans. Two case reports have recently described an infant [6] and an adult [7] with severe lung injury, whose ventilation had been facilitated by concurrent hypothermia. Since hypothermia is an accepted therapy only for HIE, we designed a preliminary translational study to investigate the effect of WBH on inflammation and surfactant status in neonates with HIE unaffected by any pulmonary injury. LETTER IN PRESS | CORRECTED PROOF 1 .

Pediatric Blood & Cancer, 2009

Pediatric Research, 2005

Objective: Immun response against pertussis can be induced by infection and/or vaccination and va... more Objective: Immun response against pertussis can be induced by infection and/or vaccination and vaccine induced immunity is known to wane within the following decade. Our aim was to assess the pertussis immun response among adolescent girls in Edirne province in Turkey and to determine its relationship with some parameters.

Journal of Hepatology, 2011

Thyroid, 2011

Resistance to thyroid hormone (RTH) is a genetic disease characterized by a reduced responsivenes... more Resistance to thyroid hormone (RTH) is a genetic disease characterized by a reduced responsiveness of the pituitary and peripheral target tissues to TH. We describe two patients with RTH in whom differentiated thyroid cancer was diagnosed. In both patients, RTH was unequivocally diagnosed and both underwent thyroidectomy for multinodular goiter. In Patient 1, histology showed a papillary thyroid carcinoma pT2. Because serum thyrotropin (TSH) levels were elevated even while the patient was taking 150 μg daily of levothyroxine (LT(4)), the patient was treated with (131)I 100 mCi for ablation of the thyroid remnant without discontinuing his LT(4) therapy. We obtained a clinically adequate response by administering LT(4) 175 μg/day (2.18 μg/kg), but the serum TSH was persistently elevated on this dose. The patient was considered "free of disease" after 8 years of follow-up. In Patient 2, histology revealed a papillary microcarcinoma (0.6 cm). Diagnostic whole-body scan was performed while the patient was taking 100 μg/day LT(4), a time that his serum TSH was 38 μU/mL. Only a small remnant was revealed, so (131)I remnant ablation was not performed. While taking LT(4) at a dose of 175 μg/day (3 μg/kg), the serum TSH was persistently high, serum TH levels were in the normal-high range, and he appeared to be clinically euthyroid. There has been no evidence of persistent or recurrent thyroid carcinoma in ultrasonography and thyroglobulin measurements that have been performed on a yearly basis for 3 years. Patients with thyroid carcinoma and RTH are a unique model of thyroid cancer in which follow-up likely occurs in the setting of constantly elevated serum TSH concentrations. The concern in these patients is that their persistent elevation of serum TSH may have an adverse effect on their thyroid cancer, and management choices in terms of the dose of LT(4) that provides the optimum lowering of serum TSH without toxicity are difficult, particularly in the situation wherein, as was the case with one of our patients, there is cardiac disease.

Thyroid, 2013

Postpartum thyroiditis (PPT) is characterized by the development of postpartum thyroid dysfunctio... more Postpartum thyroiditis (PPT) is characterized by the development of postpartum thyroid dysfunction, which may occur up to 12 months after delivery. The syndrome usually presents with transient thyrotoxicosis, followed by transient hypothyroidism. The association of this condition with resistance to thyroid hormones (RTH) has never been described. In this report, we describe a 30-year-old patient affected by RTH due to a novel p.V283A thyroid hormone receptor-β (THRB) heterozygous mutation in exon 8, which affects the ligand-binding domain, never before described in literature. A simple polymorphism was excluded through screening of 100 healthy controls. The patient became pregnant twice (in 2008 and in 2009) and developed PPT after both deliveries. Two months after her first pregnancy and one month after her second pregnancy, she presented with severe endogenous thyrotoxicosis and concomitant suppressed thyrotropin (TSH) levels, which represents an unusual finding in patients affected by RTH. Other causes of hyperthyroidism were excluded. After the hyperthyroid phase, she became hypothyroid (TSH >75 mU/L and low free-thyroxine and free-tri-iodothyronine levels), and eventually returned to her usual euthyroid status. During the course of PPT, no specific treatment was required, except for β-blockers used to treat tachycardia during the hyperthyroid phase. We report a unique case of a woman affected by RTH, due to a novel mutation V283A in THRB, who experienced PPT with a severe thyrotoxic phase after both her pregnancies. The association between RTH and PPT has never been reported in the literature. In particular, the marked suppression of TSH occurring when levels of TH are particularly elevated is not a frequent condition during RTH.

Rheumatology International, 2008

The aberrant induction of salivary/lacrimal proteins is considered to be crucial in the pathogene... more The aberrant induction of salivary/lacrimal proteins is considered to be crucial in the pathogenesis of sicca-symptoms related to primary Sjögren syndrome (SS). We report the case of an 11-year-old boy who was admitted to hospital due to recurrent bilateral parotid gland enlargement and keratoconjunctivitis, which were diagnosed as primary SS upon a combination of laboratory and instrumental tests. The proteomic analysis of the salivary peptide complex in the patient's salivary fluid near diagnosis and after 6 months of pharmacological therapy revealed quantitative and mostly qualitative differences. This observation reveals that clinical and functional changes of the salivary glands driven by non-steroidal antinflammatory drugs might be reflected in different proteomic patterns of the salivary fluid.

Platelets, 2005

Plasma homocysteine levels depend in part on the molecular nature of the methylenetetrahydrofolat... more Plasma homocysteine levels depend in part on the molecular nature of the methylenetetrahydrofolate reductase (MTHFR) and on blood folate intake. Little has been reported on platelet counts in the presence of hyperhomocysteinemia and MTHFR polymorphisms, with the exception of delayed platelet recovery in homozygous MTHFR C677T subjects after treatment with methotrexate for ovarian cancer. The aim of this investigation was to evaluate the possibility of a link between the platelet count and plasma homocysteine levels in different MTHFR variants in 165 female patients. Determinations of plasma homocysteine levels were by ELISA and of MTHFR polymorphisms (A1298C and C677T) were by inverse hybridization. Serum P- and E-selectin concentrations were obtained by ELISA. An inverse correlation (R=-0.88, P<0.001) was observed between blood platelet counts and plasma homocysteine levels in the women homozygous for MTHFR C677T. This correlation did not depend on pregnancy or other variables reported. Serum concentrations of sE- and sP-selectin, markers of endothelial and platelet activation, were significantly and positively correlated with homocysteine levels. These findings suggest that homocysteine affects platelet numbers in women with MTHFR C677T possibly consequent to endothelial and platelet activation.

Pediatric Pulmonology, 2006

ABSTRACT

Pediatric Critical Care Medicine, 2011

To verify if secretory phospholipase A2 (sPLA2) is increased in pediatric acute respiratory distr... more To verify if secretory phospholipase A2 (sPLA2) is increased in pediatric acute respiratory distress syndrome (ARDS) triggered or not by respiratory syncytial virus infection and to clarify how the enzyme may influence the disease severity and the degree of ventilatory support. Prospective pilot study. Two academic pediatric intensive care units. All infants < 6 months old hospitalized for severe respiratory syncytial virus bronchiolitis, who developed ARDS (respiratory syncytial virus-ARDS group); all infants < 6 months old diagnosed with ARDS secondary to other causes (ARDS group); and infants < 6 months old who needed ventilation for reasons other than any lung disease (control group). None. We enrolled six respiratory syncytial virus -ARDS babies, five ARDS babies, and six control infants. The sPLA2 activity and tumor necrosis factor (TNF)-α were significantly higher in the bronchoalveolar lavage of ARDS infants. Worst oxygenation, ventilation, and longer pediatric intensive care unit stay and ventilation time were present in ARDS babies. No differences were found in Clara cell secretory protein and in serum cytokines levels. Because there is no correlation between bronchoalveolar lavage protein content (a marker of permeability) and sPLA2, the enzyme seems mainly produced in the alveoli. TNF-α, the main inductor of sPLA2 expression, significantly correlates with the enzyme level in the bronchoalveolar lavage. Significant positive correlations exist between sPLA2, TNF-α and oxygen need, mean airway pressure, ventilatory index, and the Murray's lung injury score. Negative correlations were also found between sPLA2, TNF-α, and Pao2/Fio2 ratio. The sPLA2 and TNF-α are increased in ARDS and seem correlated with clinical severity, higher oxygen requirement, and more aggressive ventilation. This correlation confirms findings from adult experience and should guide further investigations on pediatric ARDS pathophysiology.

Pediatric Anesthesia, 2011

Journal of Separation Science, 2009

Listeria monocytogenes is frequently found as a contaminant in raw and ready-to-eat foods. The ab... more Listeria monocytogenes is frequently found as a contaminant in raw and ready-to-eat foods. The ability of L. monocytogenes to multiply at refrigeration temperatures and to grow in a wide range of pH values is of particular concern for food safety. According to the European Union regulation on microbiological criteria for foodstuffs, L. monocytogenes must be absent in some categories of ready-to-eat foods. The standard microbiological method for L. monocytogenes detection in foods (ISO 11290-1: 1996 (ISO, International Organization for Standardization)) is cost and time consuming. Developments of rapid, cost-effective and automated diagnostic methods to detect food-borne pathogens in foods continue to be a major concern for the industry and public health. The aim of this study was the development of a rapid, sensitive and specific molecular detection method for L. monocytogenes. To this purpose, we have applied a capillary electrophoresis method to a PCR protocol (PCR-EES (EES, experion automated electrophoresis system)) for detecting L. monocytogenes in food. In particular, a microfluidic chip-based automated electrophoresis system (experion automated electrophoresis system, Bio-Rad Laboratories, USA) was used for the rapid and automatic analysis of the amplicons. Fifty naturally contaminated samples were analysed with this method and the results were compared with those obtained with ISO method. Moreover, the microfluidic chip-based automated electrophoresis system was compared with classical gel electrophoresis (PCR-CGE). The results showed that after 24 h of culture enrichment, the PCR-EES showed a relative accuracy of 100% with ISO, while using PCR-CGE decreased it down to 96%. After 48 h of enrichment, both PCR-EES and PCR-CGE showed an accuracy of 100% with ISO.

Journal of Separation Science, 2008

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is one of the most wellknown human genetic de... more Glucose-6-phosphate dehydrogenase (G6PD) deficiency is one of the most wellknown human genetic defects, identified in more than 400 million individuals in the world. To date, no commercial kits are available for the mutation screening of this disease. Seventy G6PD-deficient Italian individuals admitted to the Laboratory of Clinical Molecular Biology of Hospital "Agostino Gemelli" of Rome were screened for the most frequent Italian mutations, by means of allele-specific PCR, followed by restriction fragment length electrophoresis. The present study compares two techniques for the identification of restriction patterns: agarose gel electrophoresis versus Experion system. When the first screening was negative, the entire G6PD gene was sequenced using the ABI 3100 Avant Instrumentation. The G6PD variants identified and their frequencies were the following: G6PD Mediterranean (75.7%), G6PD Seattle (7.1%), G6PD A -202 + 376 (7.1%), and G6PD Cassano (2.8%). In addition, we identified by direct sequencing two new mutations, namely Buenos Aires and Rignano. With the Experion method, the size band determination was more accurate than that obtained by gel electrophoresis. The Experion system resulted as a valid, easy, and reproducible diagnostic method for the screening of G6PD mutation as compared with the agarose electrophoretic analysis.

Journal of Proteome Research, 2007

A 1 year follow-up investigation of salivary acidic proline-rich proteins (aPRPs) in preterm and ... more A 1 year follow-up investigation of salivary acidic proline-rich proteins (aPRPs) in preterm and at-term newborns using HPLC-ESI-IT-MS showed that (i) this class of proteins is constitutive rather than inducible, as it is still found in the oral cavity of preterm newborns from 180 days of postconception age (PCA); (ii) the expression of PRH-2 locus anticipates that of PRH-1, since Db isoforms are expressed some months after the PRP-1 and PRP-2 isoforms. The evaluation of the relative abundances of the different aPRPs isoforms and derivatives (differently phosphorylated and cleaved) as a function of PCA showed that (iii) the proteolytic enzymes generating truncated isoforms are also constitutive because they are fully active since 180 days of PCA; (iv) the kinase involved in aPRP phosphorylation is not fully mature in preterm newborns, but its activity increases with PCA, synchronizing with that of at-term newborns and reaching the adult levels at about 500-600 days of PCA, in concomitance with the beginning of deciduous dentition.

IUBMB Life, 2009

Steroid 21-hydroxylase deficiency is present in more than 90% of patients with congenital adrenal... more Steroid 21-hydroxylase deficiency is present in more than 90% of patients with congenital adrenal hyperplasia (CAH), an inherited metabolic disorder of adrenal steroidogenesis. Impaired enzymatic activity leads to the accumulation of metabolic intermediates (progesterone and 17-hydroxyprogesterone), which results in excessive androgen production and varied signs of virilisation. CYP21A2 is an active gene and encodes for the steroid 21-hydroxylase enzyme, whereas CYP21A1P is an inactive pseudogene that contains a series of deleterious mutations. The major part of disease-causing mutations in CYP21A2 alleles are CYP21A1P-derived sequence transferred to the active gene by macro or microconversion events. Only around 5% of all disease-causing CYP21A2 alleles harbour rare mutations that do not originate from the pseudogene. In this report, we report the characterisation of two novel CYP21A2 missense mutations (p.H119R and p.I194N) found in two unrelated Italian patients with nonclassic (NC) CAH clinical diagnosis. Functional in vitro assays for mutagenized CYP21 enzymes were performed in transiently transfected mammalian cells to test the residual enzyme activity and the apparent kinetic values. The residual activities obtained for p.H119R and p.I194N mutants allowed to classify them as NC-CAH associated mutations. These results correlate with the rate of severity of the patients' disease. Finally, the new p.H119R and p.I194N mutations should be included in the panel of those already listed for association with the NC form of 21-hydroxylase deficiency. 2009 IUBMB IUBMB Life, 61(3): 229-235, 2009

Intensive Care Medicine, 2007

Mannose-binding lectin (MBL2) is a collectin molecule able to activate the complement system and ... more Mannose-binding lectin (MBL2) is a collectin molecule able to activate the complement system and the subsequent inflammatory mechanisms. Several MBL2 genetic variants have been described, including the six variants studied in this report, which are those analyzed in most detail in the medical literature. The present study analyzes the prevalence of MBL2 gene variants in preterm newborns and associates individual genotypes with pulmonary outcome variables. All polymorphisms were analyzed by means of a commercially available reverse dot-blot kit. Tertiary neonatal intensive care unit. Seventy-five consecutive preterm newborns. Two variants were particularly analyzed: -550G > C and R52C. The first one is known to be associated with lower protein synthesis when included in specific haplotypes. The homozygous and heterozygous -550G > C mutations were significantly associated with protective effects regarding different lung outcome variables, including shorter duration of mechanical ventilation, hours of continuous positive airway pressure and lower number of hemotransfusions. In contrast, the heterozygous R52C mutation was associated with unfavorable outcome, including higher bronchopulmonary dysplasia prevalence. Multivariate logistic regression analysis showed that these associations were independent of gestational age and birth weight. In addition, four groups of patients were defined on the basis of haplotype combinations. Those known to be associated with low serum MBL2 levels were linked to a better outcome in terms of factors such as hours of mechanical ventilation, continuous positive airway pressure, number of hemotransfusions and bronchopulmonary disease development. The four haplotype combination groups may have a potential diagnostic use as opposite risk factors for lung disease of prematurity.

Intensive Care Medicine, 2006

In addition to the previous classification of chronic lung disease (CLD) O2 dependency at 36 week... more In addition to the previous classification of chronic lung disease (CLD) O2 dependency at 36 weeks of postmenstrual age, a new definition of CLD has recently been proposed: new bronchopulmonary-dysplasia (BPD). This uses total duration of O2 supplementation and positive pressure requirements to delineate three degrees of severity (mild, moderate, and severe) according to the respiratory status at 36 weeks postmenstrual age. We analyzed the balance of serum proinflammatory and profibrotic/angiogenic cytokine concentrations in relation to CLD and the new BPD definition. Descriptive study in a third-level neonatal ICU. Thirty-one preterm neonates with a gestational age of 24-29 weeks were studied to evaluate their serum cytokine concentration; they were previously enrolled in a randomized clinical trial to compare the effects of high-frequency oscillatory ventilation vs. intermittent mandatory ventilation in terms of pulmonary mechanics and lung cytokines. Serum samples were collected on days 1, 3, and 5 after birth until extubation to detect the levels of three proinflammatory cytokines plus four profibrotic/angiogenic cytokines, and correlations were examined to old CLD and new BPD. Ventilation treatments were distributed homogeneously between the groups and did not interfere with the results presented here. Old CLD development, mainly corresponding to the moderate/severe forms of new BPD, was associated with increased proinflammatory and profibrotic/angiogenic cytokines, while mild forms of new BPD were characterized only by increases in profibrotic/angiogenic cytokines, suggesting a different balance of two pathogenic mechanisms in different phases of the disease.