Marco Calderisi

Contribution of a municipal solid waste incinerator to the trace metals in the surrounding soil

This study analyses the pedological environment of the area near a municipal waste incinerator that has been operating in the south-east district of Pisa for approximately 20xa0years. There are many other industrial activities in the area besides the incinerator, which represent possible sources of pollution, as well as heavy road traffic. The study area was defined by a 0–4-km zone around the site with a population of approximately 12,000 residents. The study included the physical and chemical characterisation of 100 samples of soil and an analysis of trace metals such as Cr, Ni, Pb, Zn, Hg, As and Cd. The samples were grouped into soil use categories. The results showed Zn, Pb and Hg correlated with their potentially mobile fractions, and suggested an anthropic contribution to their presence in the soil. Ni, Cr and As showed values attributable to a lithological origin. This was consistent with the PCA results. The aim was to define the environmental state of the soil of the area in order to create a reference for future research and to verify the possible presence of pollution from other sources (local industrial activities and traffic).

Vegetation at the Limits for Vegetation: Vascular Plants, Bryophytes and Lichens in a Geothermal Field

The effects of the chemical and physical factors associated with geothermal activity on plant community structure and composition were investigated in one of the largest geothermal fields of central Italy. The study site was located in the geothermal area of Sasso Pisano – Monte Rotondo Marittimo, Southern Tuscany. The percentage cover of all vascular plant, bryophyte and lichen species was estimated within 119 circular plots of 0.25xa0m2. For each plot the soil pH, soil temperature, slope, aspect, incident radiation, soil nitrogen and carbon contents were also quantified. Two vascular plants, Calluna vulgaris and Agrostis castellana, were found to be the most widespread species tolerating the harshest conditions in terms of low soil pH and high soil temperature. The most widespread cryptogam species was Hypnum cupressiforme. Spatially autoregressive models showed that a proportion of about 41–51% of the variance in species richness of one group of plants (vascular or cryptogamic plants) could be modelled by using three or four uncorrelated environmental factors respectively (soil temperature, soil nitrogen and soil C/N ratio and these three plus incident radiation). For the total number of species (vascular and cryptogamic plants), the variance explained by the same three uncorrelated variables was about 57%. This study evidenced a strong environmental control of community composition and species richness, in a site subjected to extreme soil values of soil pH and temperature. The dominance of vascular over cryptogamic vegetation in this geothermal site can be explained by the combined effects of geothermal stress (low soil pH and high soil temperature) with the summer drought typical of the Mediterranean climate.

Creatine transporter defect diagnosed by proton NMR spectroscopy in males with intellectual disability.

Creatine deficiency syndrome due to mutations in X‐linked SLC6A8 gene results in nonspecific intellectual disability (ID). Diagnosis cannot be established on clinical grounds and is often based on the assessment of brain creatine levels by magnetic resonance spectroscopy (MRS). Considering high costs of MRS and necessity of sedation, this technique cannot be used as a first level‐screening test. Likewise, gene test analysis is time consuming and not easily accessible to all laboratories. In this article feasibility of urine analysis (creatine/creatinine (Cr/Crn) ratio) performed by nuclear magnetic resonance (NMR) as a first level‐screening test is explored. Before running a systematic selection of cases a preliminary study for further molecular analysis is shown. NMR urine spectra (nu2009=u20091,347) of male patients with an ID without a clinically recognizable syndrome were measured. On the basis of abnormal Cr/Crn ratio, three patients with the highest values were selected for molecular analysis. A confirmatory second urine test was positive in two patients and diagnosis was further confirmed by a decreased brain creatine level and by SLC6A8 gene analysis. A de novo mutation was identified in one. Another patient inherited a novel mutation from the mother who also has a mild ID. A repeat urine test was negative in the third patient and accordingly creatine level in the brain and SLC6A8 gene analysis both gave a normal result. We conclude that Cr/Crn ratio measured by NMR for male patients represents a rapid and useful first level screening test preceding molecular analysis.

Using Metabolomics to Monitor Kidney Transplantation Patients by Means of Clustering to Spot Anomalous Patient Behavior

BACKGROUNDnNMR spectroscopy-based metabolomics is a system approach used to investigate the metabolic profile of biological fluids with multivariate data analysis tools. The aim of this study was to examine the kidney graft recovery process noninvasively through the examinations of urine samples using (1)H NMR spectroscopy combined with chemometric tools.nnnMETHODSnUrine samples were treated as the source of metabolites reflecting the pathological and clinical conditions of patients with transplanted kidneys. We observed 15 subjects (9 males and 6 females) during the graft recovery process and initial days thereafter. The patients provided at least 9 samples each, applying advanced statistical methods of analysis: Principal Component Analysis (PCA) and Partial Least Square Discriminant Analysis PLS-DA).nnnRESULTSnThe PCA model (for all subjects exp. var. PC1 13.96% and PC2 9.88%) allowed us to clearly designate 3 stages of recovery: initially the kidney is not working; in the second stage, it regains functions, and the third stage includes follow-up during hospitalization. PCA analysis of a single patient follows graft recovery based on biochemical (metabolites) information, assigning the appropriate recuperation stage.nnnCONCLUSIONSnNMR spectroscopy together with chemometric analysis allow monitoring of kidney graft recovery to identify patients who are not progressing within the normal range.

Predictive Role of Urinary Metabolic Profile for Abnormal MRI Score in Preterm Neonates

Background and Objective Early identification of neonates at risk for brain injury is important to start appropriate intervention. Urinary metabolomics is a source of potential, noninvasive biomarkers of brain disease. We studied the urinary metabolic profile at 2 and 10u2009days in preterm neonates with normal/mild and moderate/severe MRI abnormalities at term equivalent age. Methods Urine samples were collected at two and 10u2009days after birth in 30 extremely preterm infants and analyzed using proton magnetic resonance spectroscopy. A 3u2009T MRI was performed at term equivalent age, and images were scored for white matter (WM), cortical grey matter (cGM), deep GM, and cerebellar abnormalities. Infants were divided in two groups: normal/mild and moderately/severely abnormal MRI scores. Results No significant clustering was seen between normal/mild and moderate/severe MRI scores for all regions at both time points. The ROC curves distinguished neonates at 2 and 10u2009days who later developed a markedly less mature cGM score from the others (2u2009d: area under the curve (AUC)u2009=u20090.72, specificity (SP)u2009=u200965%, sensitivity (SE)u2009=u200975% and 10u2009d: AUCu2009=u20090.80, SPu2009=u200978%, SEu2009=u200980%) and a moderately to severely abnormal WM score (2u2009d: AUCu2009=u20090.71, specificity (SP)u2009=u200980%, sensitivity (SE)u2009=u200972% and 10u2009d: AUCu2009=u20090.69, SPu2009=u200964%, SEu2009=u200989%). Conclusions Early urinary spectra of preterm infants were able to discriminate metabolic profiles in patients with moderately/severely abnormal cGM and WM scores at term equivalent age. Urine spectra are promising for early identification of neonates at risk of brain damage and allow understanding of the pathogenesis of altered brain development.

Identification of a panel of cytokines in neonates with hypoxic ischemic encephalopathy treated with hypothermia

Purpose Inflammation is a crucial but understudied mechanism of neuronal injury after hypoxia‐ischemia. The aim was to identify a panel of cytokines involved in brain injury in neonates with hypoxic ischemic encephalopathy (HIE). Methods Ten newborns with HIE undergoing to therapeutic hypothermia (TH, HIE Group) and 8 healthy newborns (CTRL Group) were enrolled. For the HIE group, 5 samples were collected: between 0 and 6 h of life (time 1), 12 h (time 2), 24 h (time 3), 48 h (time 4) and 96 h of life (time 5). For the CTRL group, one sample was collected. A panel of 48 inflammatory cytokines was determined in all samples. Data were analyzed using multivariate statistical analysis (Principal component analysis, PCA) Results 17 cytokines, among 48 analyzed, were found to be significantly different, initially, between the CTRL and HIE groups: 12 with reported pro‐inflammatory effects and 5 with reported anti‐inflammatory effects. In the HIE group cytokines showed a decreasing trend during the TH and at the end of treatment comparable to the CTRL group. IL‐18 did demonstrate a slight increase at time 3 during HT but decreased steadily at sampling times, 4 and 5. Conclusions Our data demonstrates that many pathways of the inflammatory cascade are activated following hypoxic‐ischemic injury. This information will increase our understanding of changes in cytokines over time in neonates with HIE undergoing TH.

Predictive Role of F2-Isoprostanes as Biomarkers for Brain Damage after Neonatal Surgery

Objective Neonates have a high risk of oxidative stress during anesthetic procedures. The predictive role of oxidative stress biomarkers on the occurrence of brain injury in the perioperative period has not been reported before. Methods A prospective cohort study of patients requiring major surgery in the neonatal period was conducted. Biomarker levels of nonprotein-bound iron (NPBI) in plasma and F2-isoprostane in plasma and urine before and after surgical intervention were determined. Brain injury was assessed using postoperative MRI. Results In total, 61 neonates were included, median gestational age at 39 weeks (range 31–42) and weight at 3000 grams (1400–4400). Mild to moderate brain lesions were found in 66%. Logistic regression analysis showed a significant difference between plasma NPBI in patients with nonparenchymal injury versus no brain injury: 1.34u2009umol/L was identified as correlation threshold for nonparenchymal injury (sensitivity 67%, specificity 91%). In the multivariable analysis, correcting for GA, no other significant relation was found with the oxidative stress biomarkers and risk factors. Conclusion Oxidative stress seems to occur during anaesthesia in this cohort of neonates. Plasma nonprotein-bound iron showed to be associated with nonparenchymal injury after surgery, with values of 1.34u2009umol/L or higher. Risk factors should be elucidated in a more homogeneous patient group.