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Abstract We present a new seismotomography investigation providing a 3-D overall model of Vp, Vs and Vp/Vs for Mt. Etna, the largest and most active volcano in Europe. We estimated and jointly evaluated P- and S-wave velocity patterns together with the Vp/Vs ratio, particularly useful to discriminate the presence of groundwater, gas, and melts and thus very precious for volcano investigations. We applied the LOTOS software to ~ 4600 crustal earthquakes that occurred in the Etnean area during the last 26 years, the longest time-interval ever analysed for Mt. Etna. This wide dataset has allowed us to characterize the volcano velocity structure getting over possible singularities due to specific eruptive phases. Our results further refined the high velocity body widely recognized in the south-eastern sector of Mt. Etna by furnishing new clues on the possible former magma pathways. Moreover, the obtained 3D seismic velocity model depicted new anomalies revealing the presence of: (i) two shallow underground aquifers in the northern Etnean sector; (ii) a volume of strongly fractured rocks filled of fluids along the eastern flank; (iii) a quite deep region of probable fluid accumulation apparently not linked to the volcanic activity in the western sector.

Abstract Geophysical data provide the chance to investigate a volcano’s dynamics; considerable information can especially be gleaned on the stress and strain patterns accompanying the internal processes and the effect of magma ascent on the main structures triggering earthquakes. Here, we analysed in detail the seismicity recorded over the last two decades on Etna volcano (southern Italy), focusing on earthquakes distribution and focal mechanism clustering; the ground deformation pattern affecting the volcanic edifice with the inflation and deflation phases was also examined. Analysed data were compared in order to shed light on possible relationships with the volcanic activity and to better understand the internal dynamics of the volcano over time. Significant steps during or shortly before major eruptions in the seismic strain release and ground deformation temporal series highlight a straightforward relationship between seismicity occurring at shallow level, inflation/deflation and volcanism. Furthermore, at depths greater than 5–7 km, down to about 20 km, the orientation of the P- and T-axes clearly indicate the existence of a pressure source in the central part of the volcano. All the results underline that the stress field related to the volcano plumbing system interferes with the regional field, partly overriding it.

Abstract Seismic tomography is a very powerful and effective approach to look at depths beneath volcanic systems thus helping to better understand their behaviour. The P-wave and S-wave velocity ratio, in particular, is a key parameter useful to discriminate the presence of gas, fluids and melts. We computed the first 3-D overall model of Vp, Vs and Vp/Vs for the Lipari–Vulcano complex, central sector of the Aeolian volcanic archipelago (southern Italy). The investigated area has been characterized in recent times by fumaroles, hydrothermal activity and active degassing. In particular, in the Vulcano Island, several episodes of anomalous increases of fumarole temperature and strong degassing have been recorded in the past decades and the last “crisis”, started in September 2021, is still ongoing. For tomographic inversion we collected ~ 4400 crustal earthquakes that occurred in the last thirty years and we used the LOcal TOmography Software LOTOS. The results clearly depicted two low Vp and Vp/Vs anomalies located up to ~ 8 km depths below Vulcano and the western offshore of Lipari, respectively. These anomalies can be associated to the large presence of gas and they furnish a first picture of the gas-filled volumes feeding the main degassing activity of the area.

Due to the prominent topography of Mt. Etna, the use of satellite geodetic techniques may significantly suffer from atmospheric heterogeneities. This problem mainly affects the DInSAR technique. To overcome these drawbacks the present study attempts to make headway in measuring and interpreting atmospheric anomalies. We used the GAMIT software to obtain the ZTD (Zenith Total Delay) values for the GPS sessions performed on 1996-97, during ERS-2 passes at Mt. Etna. GAMIT software also allows to characterize the statistical behaviour of the tropospheric effects, by using residuals for each station-satellite pair, and to locate the atmospheric anomalies, present mostly at low altitudes. The attempt at using these results to produce a tomography of radio waves velocity of the troposphere suggests that the number of GPS stations used to investigate atmosphere is a critical point in such a study. The three stations are too few to invert anomalies eventually existing in the lower atmosphere. This result is a good starting point for better direct future study to verify the applicability of this tomographic technique to a geodetic network with a higher number of stations, with the aim of characterizing the lower atmosphere of Mt. Etna for a more accurate monitoring of ground deformations.

This paper analyses the ground deformations occurring on the eastern part of the Pernicana Fault from 1997 to 2005. This segment of the fault was monitored with three local networks based on GPS and EDM techniques. More than seventy GPS and EDM surveys were carried out during the considered period, in order to achieve a higher temporal detail of ground deformation affecting the structure. We report the comparisons among GPS and EDM surveys in terms of absolute horizontal displacements of each GPS benchmark and in terms of strain parameters for each GPS and EDM network. Ground deformation measurements detected a continuous left-lateral movement of the Pernicana Fault. We conclude that, on the easternmost part of the Pernicana Fault, where it branches out into two segments, the deformation is transferred entirely SE-wards by a splay fault.

Disorders of consciousness (DoC) are characterized by alteration in arousal and/or awareness commonly caused by severe brain injury. There exists a consensus on adopting advanced neuroimaging and electrophysiological procedures to improve diagnosis/prognosis of DoC patients. Currently, these procedures are prevalently applied in a research-oriented context and their translation into clinical practice is yet to come. The aim of the study consisted in the identification of measures derived from routinary electroencephalography (EEG) able to support clinicians in the prediction of DoC patients' outcome. In the present study, a routine EEG was recorded during rest from a sample of 58 DoC patients clinically diagnosed as Unresponsive Wakefulness State (UWS) and Minimally Conscious State (MCS) and followed-up for 3 months. EEG-based features characterizing brain activity in terms of spectral content and resting state networks organization were used in a predictive machine learning model to i) identify which were the most promising features in predicting patients' exit from the DoC, regardless of the clinical diagnosis and ii) verify whether such features would have been the same best discriminating UWS from MCS or specific of the outcome prediction. A predictive machine learning model was built on EEG features related to spectral content and resting state networks which returned up to 85% of performance accuracy in outcome prediction and 76% in DoC state recognition (UWS vs MCS). We provided preliminary evidence for the exploitation of a routine EEG to improve the clinical management of non-communicative patients to be confirmed in a larger DoC population.

Background/Objectives: Persons with disorders of consciousness (DoCs) may perceive pain without being able to communicate their discomfort. The Nociception Coma Scale (NCS) and its revised form (NCS-R) have been proposed to assess nociception in persons with DoCs. The main aim of this international multicenter study was to confirm (or not) our preliminary results and compare the NCS-R scores of standard stimulus (NCS-R-SS) to scores of personalized painful stimuli (NCS-R-PS). A secondary aim of the study was to verify possible correlations between the NCS-R-PS and Coma Recovery Scale-Revised (CRS-R) and to estimate convergent validity. Methods: Sixty-one patients with prolonged DoCs (pDoCs) were enrolled from seven European post-acute rehabilitation centers. Responsiveness and pain perception were assessed by CRS-R and NCS-R with standard stimulus (NCS-R-SS) and personalized stimulation (NCS-R-PS). ClinicalTrials.gov Identifier: NCT06012357. Results: our results support our prior findings on the superiority and the validity of the personalized painful stimulus approach in assessment of pain in persons with DoCs in comparison with the standardized pain assessment methodology. Conclusions : A more in-depth and tailored assessment of pain perception in persons with a DoC may lead to better acknowledgment of its presence and by extension an objective foundation for more aggressive and appropriate pain management.

Today, plastic pollution is one of the biggest threats to the environment and public health. In the tissues of exposed species, micro- and nano-fragments accumulate, leading to genotoxicity, altered metabolism, and decreased lifespan. A model to investigate the genotoxic and tumor-promoting potential of nanoplastics (NPs) is Drosophila melanogaster . Here we tested polystyrene, which is commonly used in food packaging, is not well recycled, and makes up at least 30% of landfills. In order to investigate the biological effects and carcinogenic potential of 100 µm polystyrene nanoparticles (PSNPs), we raised Oregon [R] wild-type flies on contaminated food. After prolonged exposure, fluorescent PSNPs accumulated in the gut and fat bodies. Furthermore, PSNP-fed flies showed considerable alterations in weight, developmental time, and lifespan, as well as a compromised ability to recover from starvation. Additionally, we noticed a decrease in motor activity in DNA lig4 mutants fed with PSNPs, which are known to be susceptible to dietary stressors. A qPCR molecular investigation of the larval intestines revealed a markedly elevated expression of the genes drice and p53 , suggesting a response to cell damage. Lastly, we used warts -defective mutants to assess the carcinogenic potential of PSNPs and discovered that exposed flies had more aberrant masses than untreated ones. In summary, our findings support the notion that ingested nanopolystyrene triggers metabolic and genetic modifications in the exposed organisms, eventually delaying development and accelerating death and disease.

Dysfunctions of lipid metabolism are associated with tumor progression and treatment resistance of cutaneous melanoma. BRAF/MEK inhibitor resistance is linked to alterations of melanoma lipid pathways. We evaluated whether a specific lipid pattern characterizes plasma from melanoma patients and their response to therapy. Plasma samples from patients and controls were analyzed for FASN and DHCR24 levels and lipidomic profiles. FASN and DHCR24 expression resulted in association with disease condition and related to plasma cholesterol and triglycerides in patients at different disease stages ( n = 144) as compared to controls ( n = 115). Untargeted lipidomics in plasma ( n = 40) from advanced disease patients and controls revealed altered levels of different lipids, including fatty acid derivatives and sphingolipids. Targeted lipidomics identified higher levels of dihydroceramides, ceramides, sphingomyelins, ganglioside GM3, sphingosine, sphingosine-1-phosphate, and dihydrosphingosine, saturated and unsaturated fatty acids. When melanoma patients were stratified based on a long/short-term clinical response to kinase inhibitors, differences in plasma levels were shown for saturated fatty acids (FA 16:0, FA18:0) and oleic acid (FA18:1). Our results associated altered levels of selected lipid species in plasma of melanoma patients with a more favorable prognosis. Although obtained in a small cohort, these results pave the way to lipidomic profiling for melanoma patient stratification., Competing Interests: The authors declare no conflicts of interest.

Background: Following a severe acquired brain injury, neuro-orthopaedic disorders are commonplace. While these disorders can impact patients' functional recovery and quality of life, little is known regarding the assessment, management and treatment of neuro-orthopaedic disorders in patients with disorders of consciousness (DoC)., Objective: To describe neuro-orthopaedic disorders in the context of DoC and provide insights on their management and treatment., Methods: A review of the literature was conducted focusing on neuro-orthopaedic disorders in patients with prolonged DoC., Results: Few studies have investigated the prevalence of spastic paresis in patients with prolonged DoC, which is extremely high, as well as its correlation with pain. Pilot studies exploring the effects of pharmacological treatments and physical therapy show encouraging results yet have limited efficacy. Other neuro-orthopaedic disorders, such as heterotopic ossification, are still poorly investigated., Conclusion: The literature of neuro-orthopaedic disorders in patients with prolonged DoC remains scarce, mainly focusing on spastic paresis. We recommend treating neuro-orthopaedic disorders in their early phases to prevent complications such as pain and improve patients' recovery. Additionally, this approach could enhance patients' ability to behaviourally demonstrate signs of consciousness, especially in the context of covert awareness.

The evaluation of the integrity and quantity of DNA extracted from archaeological human remains is a fundamental step before using the latest generation sequencing techniques in the study of evolutionary processes. Ancient DNA is highly fragmented and chemically modified; therefore, the present study aims to identify indices that can allow the identification of potentially amplifiable and sequenceable DNA samples, reducing failures and research costs. Ancient DNA was extracted from five human bone remains from the archaeological site of Amiternum L'Aquila, Italy dating back to the 9th-12th century and was compared with standard DNA fragmented by sonication. Given the different degradation kinetics of mitochondrial DNA compared to nuclear DNA, the mitochondrially encoded 12s RNA and 18s ribosomal RNA genes were taken into consideration; fragments of various sizes were amplified in qPCR and the size distribution was thoroughly investigated. DNA damage degree was evaluated by calculating damage frequency (λ) and the ratio between the amount of the different fragments and that of the smallest fragment (Q). The results demonstrate that both indices were found to be suitable for identifying, among the samples tested, those less damaged and suitable for post-extraction analysis; mitochondrial DNA is more damaged than nuclear, in fact, amplicons up to 152 bp and 253 bp, respectively are obtained.

The crested porcupine is a frequent host species in wildlife rescue centers and no guidelines for its management, liberation and post-liberation monitoring are yet available. Here, captive-grown porcupines' behavior and survival in the wild after liberation were investigated and described for the first time. Management strategies adopted at the centers could affect porcupine adaptation to the natural environment. The detention of porcupettes in single cages and fed only human-supplied food may not ensure ethological welfare, nor a suitable behavioral development compatible with wildlife. The liberation of captive-grown porcupines should also be carefully planned to promote and increase the possibility of post-liberation success.

Air pollution in the urban environment is a topical subject. Aero-suspended particles can cause respiratory diseases in humans, ranging from inflammation to asthma and cancer. One of the components that is most prevalent in particulate matter (PM) in urban areas is the set of tire microparticles (1-20 μm) and nanoparticles (<1 μm) that are formed due to the friction of wheels with asphalt and are increased in slow-moving areas that involve a lot of braking actions. In this work, we studied the effect that microparticles generated from passenger tires (PTWP, passenger tire wear particles) have in vitro on murine macrophages cells RAW 264.7 at two concentrations of 25 and 100 μg/mL, for 24 and 48 h. In addition to the chemical characterization of the material and morphological characterization of the treated cells by transmission electron microscopy, gene expression analysis with RT-PCR and active protein analysis with Western blotting were performed. Growth curves were obtained, and the genotoxic effect was evaluated with a comet assay. The results indicate that initially, an induction of the apoptotic process is observable, but this is subsequently reversed by Bcl2. No genotoxic damage is present, but mild cellular abnormalities were observed in the treated cells.

Humans have always been exposed to tiny particles via dust storms, volcanic ash, and other natural processes, and our bodily systems are well adapted to protect us from these potentially harmful external agents. However, technological advancement has dramatically increased the production of nanometer-sized particles or nanoparticles (NPs), and many epidemiological studies have confirmed a correlation between NP exposure and the onset of cardiovascular diseases and various cancers. Among the adverse effects on human health, in recent years, potential hazards of nanomaterials on female reproductive organs have received increasing concern. Several animal and human studies have shown that NPs can translocate to the ovary, uterus, and placenta, thus negatively impacting female reproductive potential and fetal health. However, NPs are increasingly being used for therapeutic purposes as tools capable of modifying the natural history of degenerative diseases. Here we briefly summarize the toxic effects of few but widely diffused NPs on female fertility and also the use of nanotechnologies as a new molecular approach for either specific pathological conditions, such as ovarian cancer and infertility, or the cryopreservation of gametes and embryos.

Falls are common in patients with neurological diseases and can be very problematic. Recently, there has been an increase in fall prevention research in people with neurological diseases; however, these studies are usually condition-specific (e.g., only MS, PD or stroke). Here, our aim was to evaluate and compare the efficacy of an advanced and innovative dual-task, motor-cognitive rehabilitation program in individuals with different neurological diseases who are at risk of falling. We recruited 95 consecutive adults with neurological diseases who are at risk of falling and divided them into four groups: 31 with cerebrovascular disease (CVD), 20 with Parkinson's disease (PD), 23 with traumatic brain injury (TBI) and 21 with other neurological diseases (OND). Each patient completed a dual-task, motor-cognitive training program and underwent two test evaluations to assess balance, gait, fear of falling and walking performance at the pre-and post-intervention. We found that our experimental motor-cognitive, dual-task rehabilitation program was an effective method for improving walking balance, gait, walking endurance and speed, and fear of falling, and that it reduced the risk of falls in patients with different neurological diseases. This study presents an alternative approach for people with chronic neurological diseases and provides innovative data for managing this population.

Drug resistance limits the achievement of persistent cures for the treatment of melanoma, in spite of the efficacy of the available drugs. The aim of the present study was to explore the involvement of lipid metabolism in melanoma resistance and assess the effects of its targeting in cellular models of melanoma with acquired resistance to the BRAF-inhibitor PLX4032/Vemurafenib. Since transcriptional profiles pointed to decreased cholesterol and fatty acids synthesis in resistant cells as compared to their parental counterparts, we examined lipid composition profiles of resistant cells, studied cell growth dependence on extracellular lipids, assessed the modulation of enzymes controlling the main nodes in lipid biosynthesis, and evaluated the effects of targeting Acetyl-CoA Acetyltransferase 2 (ACAT2), the first enzyme in the cholesterol synthesis pathway, and Acyl-CoA Cholesterol Acyl Transferase (ACAT/SOAT), which catalyzes the intracellular esterification of cholesterol and the formation of cholesteryl esters. We found a different lipid composition in the resistant cells, which displayed reduced saturated fatty acids (SFA), increased monounsaturated (MUFA) and polyunsaturated (PUFA), and reduced cholesteryl esters (CE) and triglycerides (TG), along with modulated expression of enzymes regulating biosynthetic nodes of the lipid metabolism. The effect of tackling lipid metabolism pathways in resistant cells was evidenced by lipid starvation, which reduced cell growth, increased sensitivity to the BRAF-inhibitor PLX4032, and induced the expression of enzymes involved in fatty acid and cholesterol metabolism. Molecular targeting of ACAT2 or pharmacological inhibition of SOAT by avasimibe showed antiproliferative effects in melanoma cell lines and a synergistic drug interaction with PLX4032, an effect associated to increased ferroptosis. Overall, our findings reveal that lipid metabolism affects melanoma sensitivity to BRAF inhibitors and that extracellular lipid availability may influence tumor cell response to treatment, a relevant finding in the frame of personalized therapy. In addition, our results indicate new candidate targets for drug combination treatments., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Vergani, Beretta, Aloisi, Costantino, Corno, Frigerio, Tinelli, Dugo, Accattatis, Granata, Arnaboldi, Rodolfo, Perego and Gatti.)

Background: Severe acquired brain injury (sABI) frequently causes impairment in self-awareness (ISA), leading to reduced patients' compliance to treatment, worse functional outcome, and high caregiver distress. Self-awareness (SA) is a multilevel and complex function that, as such, requires a specific and effective assessment. To date, many tools are available to evaluate the declarative, but not emergent and anticipatory levels of awareness, therefore the Self-Awareness Multilevel Assessment Scale (SAMAS) was recently proposed. The new tool proved to be useful to assess SA at different levels across all domains of functioning (motor, cognitive, psycho-behavioural, etc.) because it measures not only the declarative SA, but also emergent and anticipatory levels of SA, thus overcoming some important limits of other current assessment methods., Aim: This study evaluated the inter-rater reliability (IRR) of the SAMAS., Methods: Four professionals blind to each other evaluated 12 patients with sABI. Each patient was rated by two professionals., Results: Inter-rater reliability was moderate-to-excellent, adding evidence in support of the use of SAMAS to specifically diagnose ISA after sABI., Conclusions: The SAMAS can help to better address neurorehabilitation, as it allows assessing ISA as early as possible, at all possible levels of awareness and functional domains.

Objective: Social cognition can be impaired after a severe acquired brain injury (sABI), but mechanisms potentially underlying these difficulties remain to be clarified. This study aimed at investigating perspective taking ability in individuals with sABI., Method: Twenty individuals with sABI and 20 healthy controls (HCs) have been enrolled in this case-control study. All participants were submitted to an experimental visual-spatial priming memory procedure and a self-report assessment of perspective taking (i.e., the Interpersonal Reactivity Index [IRI]). Individuals with sABI were submitted to neuropsychological tests to assess executive subcomponents, working memory, and visual attention., Results: The analysis on self-report scales data documents a significant between groups difference in the IRI-Fantasy subscale, with HCs showing a higher tendency to imaginatively transpose oneself into fictional situations than individuals with sABI. Analysis of performance on the experimental procedure revealed the priming effect in HCs but not in sABI individuals. Moreover, individuals with sABI performed significantly poorer than HCs on the indices of the experimental procedure., Conclusions: Our data preliminarily demonstrated that visual-spatial perspective taking is reduced after sABI. Findings above could give some clues for the rehabilitative intervention in sABI and suggest the possible application of the procedure here used in assessing perspective taking after sABI., (© The Author(s) 2021. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Pain and suffering in persons with disorders of consciousness (DoC) remain poorly understood, frequently unaddressed or inadequately addressed, and controversial on numerous levels. This narrative literature review will address a number of critical issues germane to pain and suffering in this challenging group of patients, providing an introductory overview of the topic, perspectives on current knowledge regarding pain pathoanatomy and pathophysiology, and a review of common pain generators and factors that can lead to the chronifcation of pain. Caveats on bedside pain assessment challenges, as well as electrophysiologic and neuroimaging findings in these patients, will also be explored. Pain management techniques, including non-pharmacological and pharmacological, will be reviewed. Ethical considerations in the context of pain and suffering in persons with disorders of consciousness will round out the review prior to our concluding comments.

Indoor air pollutants (IAP), which can pose a serious risk to human health, include biological pollutants, nitric oxide (NO), nitrogen dioxide (NO 2 ), volatile organic compounds (VOC), sulfur dioxide (SO 2 ), carbon monoxide (CO), carbon dioxide (CO 2 ), silica, metals, radon, and particulate matter (PM). The aim of our work is to conduct a multidisciplinary study of fine silica particles (<2.5 μm) in the presence or absence of ozone (O 3 ), and evaluate their potential cytotoxicity using MTS, micronucleus, and the comet test in two cell lines. We analyzed A549 (human basal alveolar epithelial cell adenocarcinoma) and Hs27 (human normal fibroblasts) exposed to dynamic conditions by an IRC simulator under ozone flow (120 ppb) and in the presence of silica particles (40 μg/h). The viability of A549 and Hs27 cells at 48 and 72 h of exposure to silica or silica/ozone decreases, except at 72 h in Hs27 treated with silica/ozone. The micronucleus and comet tests showed a significant increase in the number of micronuclei and the % of DNA in the queue, compared to the control, in both lines in all treatments, even if in different cell times/types. We found that silica alone or with more O 3 causes more pronounced genotoxic effects in A549 tumor cells than in normal Hs27 fibroblasts.

Animal-vehicle collisions are the major cause of rescue and need for hospitalization in wildlife referral centers. Clinicians need easy-to-use tools to make rapid decisions about whether to euthanize or treat injured animals. The aim of the study was to evaluate the data (reasons for rescue, diagnosed lesions, and outcome) from a hospitalized population of wildlife ungulates and investigate the benefits of using radiography as a diagnostic tool. Data were collected from three wildlife referral centers in Tuscany (Italy). The following information was collected for each animal: reason for hospitalization, clinical examination, radiographic examination, definitive diagnosis, and outcome. A chi-squared test was used to assess the benefits of radiography in detecting different traumatic lesions. Prevalence was reported according to the reason for hospitalization, definitive diagnosis, radiographic diagnosis, and outcome. The main reason for hospitalization was traumatic lesions due to vehicle collisions and 71.1% of the animals did not survive. Radiography was more useful in patients with traumatic axial skeletal lesions and/or multiple traumas with respect to traumatic appendicular skeleton lesions. Our results show that radiography is a useful diagnostic technique for assessing wildlife emergencies and it could help the clinician in making medical decisions.

The potential risks of environmental nanoparticles (NPs), in particular Polystyrene Nanoparticles (PNPs), is an emerging problem; specifically, the interaction of PNPs with intestinal cells has not been characterized so far. The mechanism by which polystyrene particles are transferred to humans has not yet been clarified, whether directly through ingestion from contaminated food. We evaluated the interaction between PNPs and colorectal adenocarcinoma cells (HCT116). Cells were exposed to different concentrations of PNPs, metabolic activity and the consequent cytotoxic potential were assessed through viability test; we evaluated the PNP genotoxic potential through the Cytokinesis-Block Micronucleus cytome (CBMN cyt) assay. Finally, we detected Reactive Oxygen Species (ROS) production after NPs exposure and performed Western Blot analysis to analyze the enzymes (SOD1, SOD2, Catalase, Glutathione Peroxidase) involved in the cell detoxification process that comes into play during the cell-PNPs interaction. This work analyzes the cyto and genotoxicity of PNPs in the colorectal HCT116 cell line, in particular the potential damage from oxidative stress produced by PNPs inside the cells related to the consequent nuclear damage. Our results show moderate toxicity of PNPs both in terms of ROS production and DNA damage. Further studies will be needed on different cell lines to have a more complete picture of the impact of environmental pollution on human health in terms of PNPs cytotoxicity and genotoxicity., Competing Interests: The authors have declared that no competing interests exist.

Introduction: Persons with disorders of consciousness (DoC) may perceive pain without being able to communicate their discomfort. Nociception Coma Scale (NCS) and its revised form (NCS-R) have been proposed to assess nociception in coma survivors with DoC., Objective: Aim of the present study was to compare, in non-communicative patients with DoC, NCS-R scores obtained with the standard pressure on fingernail bed (standard stimulus, SS) versus other personalized painful stimuli (PS), to verify possible correlations between NCS-R and Coma Recovery Scale-Revised (CRS-R)., Materials and Methods: Twenty-one patients with DoC were included in the study. Responsiveness and pain perception were assessed by CRS-R and NCS-R with standard stimulus (NCS-R-SS) and personalized stimulation (NCS-R-PS). Statistical analysis was performed with the nonparametric Wilcoxon test for comparison of both total NCS-R-SS and NCS-R-PS scores., Results: NCS-R at admission showed that 9 of 21 patients (42.8%) had higher scores in response to personalized stimulus compared to standard stimulus. Significant correlation with CRS-R were found for both NCS-R-SS ( R  = 0.701, p  = .008) and NCS-R-PS ( R  = 0.564, p  = .045). Discussion: The preliminary results obtained in the present study suggest that NCS-R-PS may disclose pain perception in a larger number of non-communicative patients with DoC, compared to NCS-R-SS.

The crested porcupine is a widespread naturalized Italian rodent of African origin; nevertheless, very little information on the population abundance and its health status is available. In this study, the hematological and serum chemistry profile of 10 free-ranging captured crested porcupines was established for the first time. The mean hematological values resulted: 5.7 SD 0.4 M/μL for red blood cells; 13.6 SD 0.8 g/dL for hemoglobin; 77.3 SD 5.7 fL for mean corpuscular volume and 30.1 SD 4.7 g/dL for mean corpuscular hemoglobin concentration; 14.4 SD 7.2 K/μL for white blood cells; and 557.0 SD 469.9 K/μL for platelets. The mean urea and creatinine values resulted with 19.8 SD 8.3 mg/dL and 1.6 SD 3.0 mg/dL, respectively. The mean value of total protein was 6.7 SD 1.0 g/dL, with values of albumin higher than globulins. The mean activity of creatine kinase, aspartate transaminase, gamma-glutamyl transpeptidase, and alkaline phosphatase was 927.3 SD 607.6 U/L, 199.2 SD 70.8 U/L, 16.9 SD 13.7 U/L, and 256 SD 75.8 U/L, respectively. Highest values of alkaline phosphatase were recorded in two porcupines presenting severe injuries with clear signs of infection. These preliminary results may be a helpful tool in order to assess porcupine health status.

Several studies have provided information on environmental nanoplastic particles/debris, but the in vitro cyto-genotoxicity is still insufficiently characterized. The aim of this study is to analyze the effects of polystyrene nanoparticles (PNPs) in the Hs27 cell line. The viability of Hs27 cells was determined following exposure at different time windows and PNP concentrations. The genotoxic effects of the PNPs were evaluated by the cytokinesis-block micronucleus (CBMN) assay after exposure to PNPs. We performed ROS analysis on HS27 cells to detect reactive oxygen species at different times and treatments in the presence of PNPs alone and PNPs added to the Crocus sativus L. extract. The different parameters of the CBMN test showed DNA damage, resulting in the increased formation of micronuclei and nuclear buds. We noted a greater increase in ROS production in the short treatment times, in contrast, PNPs added to Crocus sativus showed the ability to extract, thus reducing ROS production. Finally, the SEM-EDX analysis showed a three-dimensional structure of the PNPs with an elemental composition given by C and O. This work defines PNP toxicity resulting in DNA damage and underlines the emerging problem of polystyrene nanoparticles, which extends transversely from the environment to humans; further studies are needed to clarify the internalization process.

The Coma Recovery Scale-Revised (CRS-R) is the gold standard of responsiveness assessment in patients with disorder of consciousness. The purpose of this study is to search for the efficacy of the caregivers' involvement in the evaluation of responsiveness in these patients. Responsiveness assessment was performed in 15 patients with CRS-R. The CRS-R was administered with and without the emotional stimulation of the primary caregiver at different times. Our preliminary findings seem to suggest that, including also the caregivers during CRS-R assessment, may obtain better responsiveness scoring than that obtained by professionals and might reduce the misdiagnosis rate.

Patients with a prolonged disorder of consciousness (DoC) may present with severe spasticity and diffuse pain, which might impair motor output, thus preventing any possible behavioral responsiveness. A 26-year-old man affected by frontoparietal hemorrhage was operated by hematoma evacuation and decompressive craniectomy; coma persisted for 1 month; cranioplasty and ventriculo-peritoneal shunting was performed after 4 months. At admission in rehabilitation, he was diagnosed as vegetative state/unresponsive wakefulness syndrome (VS/UWS). The implantation of intrathecal baclofen (ITB) pump (Medtronic SynchroMed™ II), 14 months after, (60 μg/daily), dramatically improved behavioral responsiveness according to Coma Recovery Scale-Revised (CRS-R) from 6 to 12 (1 month after ITB). Nociception Coma Scale-Revised (NCS-R) also changed from 4 to 8 at the same time points. This case report may be an example of covert cognition that should have been diagnosed as a functional locked-in syndrome or motor-cognitive dissociation, rather than as VS/UWS.

Background. Language disorders may occur in patients with disorders of consciousness (DoCs), and they could interfere with the behavioral assessment of consciousness and responsiveness. Objective. In this study, we retrospectively explored whether ERP N400 was eventually associated with the presence of aphasia diagnosed in those patients who had evolved into Exit-Minimally Conscious State (E-MCS) at the clinical follow-up. Methods. In this retrospective cohort study, the ERPs elicited by an auditory sentences task were retrospectively examined in a sample of 15 DoC patients diagnosed according to the Coma Recovery Scale-Revised (CRS-R). All these 15 DoC patients underwent a (at least) 1-year clinical follow-up, which included a neuropsychological evaluation to assess language function among other cognitive functions. Ten healthy individuals also underwent the same paradigm to investigate the variability of ERPs characteristics. Results. The N400 ERP component with centroparietal topography was found in 9 of 10 healthy controls in response to the ill-formed sentences. Analysis of patients' data revealed that (1) a significant N400 component could be detected in 64% (9 of 14 patients) of the DoC patients regardless of the type of DoC; (2) no significant N400 ERP component was retrospectively detected in those E-MCS patients who showed aphasia at the follow-up; and (3) the presence/absence of the N400-ERP component was consistent with the brain lesion side and significantly predict the recovery. Conclusion. These preliminary findings indicate that the absence of N400 was significantly associated with the presence of aphasia diagnosed at the clinical follow-up in E-MCS patients.

There has been a significant evolution of nomenclature with regards to classification of persons with disorders of consciousness (DoC) over the last 100 years. This paper provides a review of the evolution of this terminology with discussion of the advantages and disadvantages of historical and current terms. Recommendations for how this evolution should continue moving forward in the best interest of patients, their families, society, clinical care, and research will also be addressed. The taxonomy we choose, hopefully by international consensus, has multifaceted implications that go well beyond just a debate on nomenclature.

To date, no international guidelines or recommendations for diagnosis or prognosis of patients with disorders of consciousness (DoC) have been established. The International Brain Injury Association's (IBIA) Special Interest Group on Disorders of Consciousness (DoC-SIG) launched an international multicenter survey to compare diagnostic and prognostic procedures across countries and clinical settings. Objectives : To explore which specific diagnostic protocols and prognostic indices were utilized in the care for persons with DoC in different countries and to determine the usage, if any, of national guidelines in the care of such patients. Methods : The questionnaire included 17 questions in two distinct sections (I - clinical and instrumental tools and involvement of caregivers and II - clinical, anamnestic and instrumental markers). Results : Physicians composed 50% of the survey respondents (120) and were all involved in post-acute rehabilitation care. In the majority of countries, respondents reported that there were no national guidelines or recommendations for DoC care. The Glasgow Coma Scale (GCS) and the Coma Recovery Scale-Revised (CRS-R) were the most frequently used clinical scales for diagnostic purposes. The majority of respondents reported the involvement of caregivers in the evaluation of behavioral responsiveness of patient with DoC. The survey indicated that only a few centers performed neurophysiological investigations routinely as diagnostic instrumental procedures. Our results suggest that international guidelines and recommendations for the care of persons with DoC still need to be formulated and ideally agreed to by consensus.

Primary Objective : The primary aim of this study was to determine the frequency of severe impaired self-awareness (ISA) in patients with severe traumatic brain injury (TBI) and the correlates of selected clinical, neuropsychiatric and cognitive variables. The secondary aim of the study was to assess depression and apathy on the basis of their level of self-awareness. Methods : Thirty patients with severe TBI and 30 demographically matched healthy control subjects (HCs) were compared on measures of ISA, depression, anxiety, alexithymia, neuropsychiatric symptoms and cognitive flexibility. Results : Twenty percent of the patients demonstrated severe ISA. Severe post-acute ISA was associated with more severe cognitive inflexibility, despite the absence of differences in TBI severity, as evidenced by a Glasgow Coma Scale (GCS) score lower than 9 in all cases in the acute phase. Patients with severe ISA showed lower levels of depression and anxiety but tended to show more apathy and to have greater difficulty describing their emotional state than patients with severe TBI who showed minimal or no disturbance in self-awareness. Conclusion : These findings support the general hypothesis that severe ISA following severe TBI is typically not associated with depression and anxiety, but rather with apathy and cognitive inflexibility.

Introduction: The Brief Post-Coma Scale (BPCS) is an easy diagnostic tool for individuals with disorders of consciousness (DoC), in a reduced version from a previously Post-Coma Scale, that could distinguish patients in the minimally conscious state (MCS) from those in unresponsive wakefulness syndrome (UWS), formerly defined as vegetative state (VS)., Objective: Aim of the study was to assess the diagnostic validity of the BPCS in comparison with the Coma Recovery Scale-Revised (CRS-R), in its Italian validated version, the Disability Rating Scale (DRS), the Level of Cognitive Functioning (LCF), and the Glasgow Outcome Scale (GOS)., Methods: In an Italian multicenter study on 545 patients with DoC, 36 post-acute rehabilitation wards, 32 long-term care centers, and 2 family associations participated to data collection., Results: Statistically significant correlations were found between the BPCS and the other clinical scales: R = 0.586 (p < 0.001) with LCF, R = - 0.566 (p < 0.001) with DRS, R = 0.622 (p < 0.001) with CRS-R. The BPCS scores resulted significantly correlated with the time from acute event (R = 0.117, p = 0.006). Patients with GOS score 2 had mean BPCS of 1.84 ± 1.19, whereas those with GOS 3 had significantly higher scores 3.88 ± 1.71 (p < 0.001). Similarly, in patients with vegetative state/UWS (VS/UWS), the mean BPSC score was 1.71 ± 1.09, significantly lower (p < 0.001) than that of patients with minimally conscious state (BPCS = 3.83 ± 1.29). Finally, the agreement of the BPCS and clinical diagnosis was of 84.4%, with an odds ratio OR = 3.781 (95% CI = 3.026-4.725, p < 0.001)., Conclusions: The BPCS has demonstrated statistically significant correlations with the most commonly used scales in persons with DoC.

Introduction: Severe acquired brain injury (sABI) is considered the most common cause of death and disability worldwide. sABI patients are supported by their caregivers who often exhibit high rates of psychological distress, mood disorders, and changes in relationship dynamics and family roles., Objectives: To explore lifestyle changes of caregivers of sABI patients during the postacute rehabilitation, by investigating possible differences between primary and secondary caregivers. Primary caregivers spend most of the time with the patient, providing daily care and taking most responsibility for the day-to-day decisions, while secondary caregivers are those who provide additional support., Methods: Three hundred forty-seven caregivers of sABI patients were asked to fill in an unpublished self-report questionnaire to explore their possible lifestyles changes., Results: A statistically significant difference was found between primary and secondary caregivers in time spent in informal caregiving (p<0.001). The primary caregivers reduced all leisure activities compared to secondary carers (p<0.05)., Conclusions: By comparing the percentage of leisure activities performed by caregivers before and after the patient's sABI onset, all caregivers showed high percentages of changes in lifestyle and habits, even though primary caregivers reported more negative lifestyle changes than secondary caregivers. Further studies are needed to investigate needs and burden experienced by caregivers of sABI patients during the postacute rehabilitation phase, also in relation to the patients' outcome, to address support interventions for them and improve their quality of life.

Patients with severe acquired brain injury (SABI) may evolve towards different outcomes. The primary aim was to evaluate the clinical evolution of a large population of patients with SABI admitted to post-acute rehabilitation from 2001 to 2016, diagnosed with severe brain injury (GCS ≤ 8) in the acute phase and a coma duration of at least 24 h. The possible changes between the admission time to a post-acute rehabilitation hospital and the discharge time were measured by means of Glasgow Outcome Scale (GOS), Level of Cognitive Functioning (LCF), and Disability Rating Scale (DRS). We also correlated the improvement rate with some sociodemographic and clinical features of the individuals with SABI enrolled. Data of 890 patients were analyzed (54% TBI, length of stay = 162 ± 186 days, GCS = 7.46 ± 1.28); time interval from the SABI (OR = 0.246, CI 95% = 0.181 - 0.333), scores at admission of LCF (OR = 2.243, CI 95% = 1.492 - 3.73), GOS (OR = 0.138, CI 95% = 0.071 - 0.266), DRS (OR = 0.457, CI 95% = 0.330 - 0.632), and etiology (OR = 2.273, CI 95% = 1.676 - 3.084) played a significant role (p < 0.001, explained variance 69.9%) for improving GOS score. Time interval from the SABI to admission in our post-acute rehabilitation ward (OR = 0.300, CI 95% = 0.179 - 0.501, p < 0.001), length of rehabilitation stay (OR = 2.808, CI 95% = 1.694 - 4.653, p < 0.001), and etiology (OR = 1.769, CI 95% = 1.095 - 2.857, p = 0.020) led to a statistically significant improvement in DRS (explained variance 91%). The most significant predictive factors for the outcome of patients with SABI were etiology, time interval from SABI to admission in rehabilitation, and length of rehabilitation stay.

In this paper we present a new analytical GC/MS method for the analysis of mixtures of free fatty acids and metal soaps in paint samples. This approach is based on the use of two different silylating agents: N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) and 1,1,1,3,3,3-hexamethyldisilazane (HMDS). Our experimentation demonstrated that HMDS does not silylate fatty acid carboxylates, so it can be used for the selective derivatization and GC/MS quantitative analysis of free fatty acids. On the other hand BSTFA is able to silylate both free fatty acids and fatty acids carboxylates. The reaction conditions for the derivatization of carboxylates with BSTFA were thus optimized with a full factorial 3 2 experimental design using lead stearate and lead palmitate as model systems. The analytical method was validated following the ICH guidelines. The method allows the qualitative and quantitative analysis of fatty acid carboxylates of sodium, calcium, magnesium, aluminium, manganese, cobalt, copper, zinc, cadmium, and lead and of lead azelate. In order to exploit the performances of the new analytical method, samples collected from two reference paint layers, from a gilded 16th century marble sculpture, and from a paint tube belonging to the atelier of Edvard Munch, used in the last period of his life (1916-1944), were characterized., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Retrograde amnesia (RA), which includes loss of memory for past personal events (autobiographical RA) and for acquired knowledge (semantic RA), has been largely documented in patients with amnestic mild cognitive impairment (aMCI). However, previous studies have produced controversial results particularly concerning the temporal extent of memory impairment. Here we investigated whether, with the onset of hippocampal pathology, age of memory acquisition and retrieval frequency play different roles in modulating the progressive loss of semantic and episodic contents of retrograde memory respectively. For this purpose, aMCI patients and healthy controls were tested for the ability to recall semantic and autobiographical information related to famous public events as a function of both age of acquisition and retrieval frequency. In aMCI patients, we found that the impairment in recollecting past personal incidents was modulated by the combined action of memory age and retrieval frequency, because older and more frequently retrieved episodes are less susceptible to loss than more recent and less frequently retrieved ones. On the other side, we found that the loss of semantic information depended only on memory age, because the remoteness of the trace allows for better preservation of the memory. Our results provide evidence that the loss of the two components of retrograde memory is regulated by different mechanisms. This supports the view that diverse neural mechanisms are involved in episodic and semantic memory trace storage and retrieval, as postulated by the Multiple Trace Theory.

Background: The Encephalomyocarditis virus (EMCV) is a small, non enveloped, positive sense single-stranded RNA virus in the genus Cardiovirus, family Picornaviridae, with two known serotypes. It is spread worldwide and infects a huge range of vertebrate hosts with zoonotic potential for humans. The pig is the mammal most likely to be impacted on with the disease, but EMCV occurrence has also been reported in non-human primates and in a variety of domestic, captive and wild animals. Until now, human cases have been very rare and the risk appears to be almost negligible in spite of human susceptibility to the infection., Case Presentation: Between September and November 2012 a fatal Encephalomyocarditis virus outbreak involving four Barbary macaques and 24 crested porcupines occurred at a rescue centre for wild and exotic animals in Central Italy. In this open-field zoo park located near Grosseto, Tuscany about 1000 animals belonging to different species, including various non-human primates were hosted at that time. Sudden deaths were generally observed without any evident symptoms or only with mild nonspecific clinical signs. The major gross change was characterised by grey-white necrotic foci in the myocardium and the same EMCV strain was isolated both in macaques and crested porcupines. Phylogenetic analysis has confirmed that only one EMCV strain is circulating in Italy, capable of infecting different animal species., Conclusions: This report confirms the susceptibility of non-human primates to the EMCV infection and describes the disease in porcupine, a common wild Italian and African species. No human cases were observed, but given the zoonotic potential of EMCV these findings are of importance in the context of animal-human interface.

We developed a functional semi-structured scale to observe Hemineglect symptoms in Activities of Daily Living (H-ADL). The scale could assist clinicians in assessing rehabilitation priorities aimed at correcting any persisting errors or omissions. In addition, the scale could also be used by caregivers to observe patients' progress and improve their participation. Two groups of right brain-damaged patients (25 with hemineglect; 27 without hemineglect) were tested twice: at admission and before discharge from hospital. A control group of healthy individuals matched to patients for age and education and patients' caregivers also participated. Two raters (A; B), experts in neuropsychology, observed patients and healthy individuals using the H-ADL. We found that the H-ADL final scores correlated with the standard hemineglect tests. The three groups differed in performance and differences also emerged between the first and the second assessment, suggesting an improvement due to the remission of hemineglect as a consequence of the treatment. Raters A and B did not differ in their observations, but there were some discrepancies with caregivers' observations. Therefore, although caregivers could help clinicians in detecting persistent hemineglect behaviour, the assessment should be performed by experts in neuropsychology.