Study and development of a battery of tests for the neuropsychological evaluation of imputability according to the cognitive approach

openNel Codice di Procedure Penale Italiano (Codice Rocco del 1930 ed attualmente in vigore), l’Articolo 85 definisce chi, alla conclusione del processo, può essere ritenuto imputabile del reato ascritto qualora al momento del fatto fosse ritenuto capace di intendere e di volere; l’Articolo in questione recita: “Nessuno può essere punito per un fatto preveduto dalla legge come reato, se, al momento in cui lo ha commesso, non era imputabile. È imputabile chi ha la capacità di intendere e volere” (art. 85 c.p.p.). Questa definizione lascia intendere pertanto che non è sufficiente accertarsi che l’imputato abbia materialmente messo in atto il comportamento criminoso perché lo si possa ritenere responsabile di quanto commesso, ma serve capire se nel metterlo in opera abbia compreso gli elementi salienti della situazione, il significato del proprio agito e le conseguenze alla quali tale agito porta - definendo quindi la capacità di intendere - e che quanto fatto sia frutto di una pianificazione intenzionale, considerando la situazione, l’adeguamento delle proprie azioni alle intenzioni e che tali azioni siano cognitivamente controllate nel mentre si svolgono – riferendosi quindi alla capacità di volere (Gulotta, 2020). Sebbene il concetto di imputabilità sia a tutti gli effetti appartenete al mondo giuridico, esso trova comunque ampio spazio nella ricerca scientifica in ambito psicologico e psichiatrico, nello studio di quelle funzioni cognitive che sottostanno e determinano il funzionamento psicologico atipico dovuto, ad esempio alla presenza di una psicopatologia o patologia organica che, previo accertamento di una relazione sussistente tra questi due fenomeni, definito “nesso causale”, che collega appunto il comportamento oggetto di valutazione e la patologia stessa, può fornire un’indicazione circa l’imputabilità del soggetto. Essenzialmente ciò significa che non basta accertarsi della presenza di deficit mentale od organico affinché si possa ravvisare un difetto della capacità di intendere e di volere, ma tale mancanza deve incidere sulle funzioni cognitive relative alla capacità d’intendere e di volere, che sono alla base del concetto di imputabilità. Con questo elaborato s’intende porre l’attenzione sulle modalità di accertamento di tale capacità, ad oggi ravvisabile sostanzialmente tramite la metodologia d’indagine del colloquio clinico psichiatrico, con una scarsa considerazione di strumenti valutativi che adeguatamente svolgono un ruolo di supporto al professionista nell’aiutarlo a prendere una decisione circa i vari casi seguiti. Nel determinare quali sono le specifiche funzioni d’indagine, si andranno ad individuare le principali funzioni cognitive sottostanti i comportamenti e le manifestazioni che giustificano un determinato stato mentale, di specifico interesse forense, secondo dunque un approccio di tipo cognitivo. Come verrà approfondito più avanti, uno dei maggiori difetti del colloquio clinico come metodo d’indagine della psicopatologia in ambito forense consiste in una bassa concordanza tra diversi valutatori circa lo stesso oggetto d’indagine (Conti, 2008), ossia una bassa inter-rater reliability. Il punto della questione, dunque, consiste nel fornire uno strumento affidabile che permetta una maggiore concordanza nei giudizi tra gli psicologi, i quali andranno ad esprimere la loro valutazione dovendo considerare lo strumento utilizzato. Lo strumento in questione consiste in una batteria di test (o reattivi) di tipo neuropsicologico che valuta le funzioni cognitive sottostanti la capacità di intendere e volere, fornendone una valutazione oggettiva utilizzabile nel contesto forense; vedremo nei capitoli successivi i costrutti (funzioni cognitive) indagati, i motivi per cui si sono scelte, la metodologia utilizzata ed i risultati ottenuti

Correction of distortions in MR Echo Planar images using a super-resolution T2-Weighted volume

Magnetic resonance imaging (MRI) is a widely used technique to assess brain diseases without the use of ionizing radiations. Brain anatomy can be captured using T1-Weighted (T1W) and T2-Weighted (T2W) acquisitions. In addition to mapping brain anatomy, MRI can be also applied to study the brain functions through a process called the hemodynamic response. Blood releases oxygen to neurons at a greater rate than to inactive neurons: this causes a change of the relative levels of oxygenated and deoxygenated blood, i.e. a change of the contrast between the two level of blood oxygenation that can be detected on the basis of their differential magnetic susceptibility. This acquisition technique is called functional Magnetic Resonance Imaging (fMRI), and it represents an indirect measure of the neuron activity. Although BOLD-based techniques have been shown to work reliably for a huge range of applications, straight-forward BOLD imaging has some inherent problems (such as macroscopic field inhomogeneity effects that produce spatial distortions in the acquisitions). The aim of this thesis is to give an overview of the fMRI data analysis focusing on some aspects of the preprocessing pipeline. In chapter 1 we will introduce the problem of Echo Planar Imaging (EPI) spatial distortions and a new method to correct them, based on non-linear registrations to an intra-subject T2W volume. In chapter 2 we will show the procedure for the construction of a good reference to apply the EPI-distortions correction method. This method belongs to the super-resolution algorithms and it aims to produce a T2W high resolution reference. In chapter 3, the previous methods will be combined together to perform the EPI distortion correction method. Finally, in chapter 4 we will present a bunch of clinical fMRI studies where the correction method was performed. Our results provide a good evidence of the effectiveness of the combined approach, which gives the advantage of using only standard acquisition protocol to have alle the information required to perform the proposed EPI-distortion correction

Effects of airborne pollutants on mitochondrial DNA Methylation

Background: Mitochondria have small mitochondrial DNA (mtDNA) molecules independent from the nuclear DNA, a separate epigenetic machinery that generates mtDNA methylation, and are primary sources of oxidative-stress generation in response to exogenous environments. However, no study has yet investigated whether mitochondrial DNA methylation is sensitive to pro-oxidant environmental exposures. Methods: We sampled 40 male participants (20 high-, 20 low-exposure) from each of three studies on airborne pollutants, including investigations of steel workers exposed to metal-rich particulate matter (measured as PM1) in Brescia, Italy (Study 1); gas-station attendants exposed to air benzene in Milan, Italy (Study 2); and truck drivers exposed to traffic-derived Elemental Carbon (EC) in Beijing, China (Study 3). We have measured DNA methylation from buffy coats of the participants. We measured methylation by bisulfite-Pyrosequencing in three mtDNA regions, i.e., the transfer RNA phenylalanine (MT-TF), 12S ribosomal RNA (MT-RNR1) gene and “D-loop” control region. All analyses were adjusted for age and smoking. Results: In Study 1, participants with high metal-rich PM1 exposure showed higher MT-TF and MT-RNR1 methylation than low-exposed controls (difference = 1.41, P = 0.002); MT-TF and MT-RNR1 methylation was significantly associated with PM1 exposure (beta = 1.35, P = 0.025); and MT-RNR1 methylation was positively correlated with mtDNA copy number (r = 0.36; P = 0.02). D-loop methylation was not associated with PM1 exposure. We found no effects on mtDNA methylation from air benzene (Study 2) and traffic-derived EC exposure (Study 3). Conclusions: Mitochondrial MT-TF and MT-RNR1 DNA methylation was associated with metal-rich PM1 exposure and mtDNA copy number. Our results suggest that locus-specific mtDNA methylation is correlated to selected exposures and mtDNA damage. Larger studies are needed to validate our observations

Functional and Structural Brain Damage in Friedreich's Ataxia

Friedreich's ataxia (FRDA) is a rare hereditary neurodegenerative disorder caused by a GAA repeat expansion in the FXN gene. There is still no cure or quantitative biomarkers reliaby correlating with the progression rate and disease severity. Investigation of functional and structural alterations characterizing white (WM) and gray matter (GM) in FRDA are needed prerequisite to monitor progression and response to treatment. Here we report the results of a multimodal cross-sectional MRI study of FRDA including Voxel-Based Morphometry (VBM), diffusion-tensor imaging (DTI), functional MRI (fMRI), and a correlation analysis with clinical severity scores. Twenty-one early-onset FRDA patients and 18 age-matched healthy controls (HCs) were imaged at 3T. All patients underwent a complete cognitive and clinical assessment with ataxia scales. VBM analysis showed GM volume reduction in FRDA compared to HCs bilaterally in lobules V, VI, VIII (L>R), as well as in the crus of cerebellum, posterior lobe of the vermis, in the flocculi and in the left tonsil. Voxel-wise DTI analysis showed a diffuse fractional anisotropy reduction and mean, radial, axial (AD) diffusivity increase in both infratentorial and supratentorial WM. ROI-based analysis confirmed the results showing differences of the same DTI metrics in cortico-spinal-tracts, forceps major, corpus callosum, posterior thalamic radiations, cerebellar penduncles. Additionally, we observed increased AD in superior (SCP) and middle cerebellar peduncles. The WM findings correlated with age at onset (AAO), short-allelle GAA, and disease severity. The intragroup analysis of fMRI data from right-handed 14 FRDA and 15 HCs showed similar findings in both groups, including activation in M1, insula and superior cerebellar hemisphere (lobules V–VIII). Significant differences emerged only during the non-dominant hand movement, with HCs showing a stronger activation in the left superior cerebellar hemisphere compared to FRDA. Significant correlations were found between AAO and the fMRI activation in cerebellar anterior and posterior lobes, insula and temporal lobe. Our multimodal neuroimaging protocol suggests that MRI is a useful tool to document the extension of the neurological impairment in FRDA

Vascular endothelial growth factor receptor 3 signaling contributes to angioobliterative pulmonary hypertension

The mechanisms involved in the development of severe angioobliterative pulmonary arterial hypertension (PAH) are multicellular and complex. Many of the features of human severe PAH, including angioobliteration, lung perivascular inflammation, and right heart failure, are reproduced in the Sugen 5416/chronic hypoxia (SuHx) rat model. Here we address, at first glance, the confusing and paradoxical aspect of the model, namely, that treatment of rats with the antiangiogenic vascular endothelial growth factor (VEGF) receptor 1 and 2 kinase inhibitor, Sugen 5416, when combined with chronic hypoxia, causes angioproliferative pulmonary vascular disease. We postulated that signaling through the unblocked VEGF receptor VEGFR3 (or flt4) could account for some of the pulmonary arteriolar lumen-occluding cell growth. We also considered that Sugen 5416-induced VEGFR1 and VEGFR2 blockade could alter the expression pattern of VEGF isoform proteins. Indeed, in the lungs of SuHx rats we found increased expression of the ligand proteins VEGF-C and VEGF-D as well as enhanced expression of the VEGFR3 protein. In contrast, in the failing right ventricle of SuHx rats there was a profound decrease in the expression of VEGF-B and VEGF-D in addition to the previously described reduction in VEGF-A expression. MAZ51, an inhibitor of VEGFR3 phosphorylation and VEGFR3 signaling, largely prevented the development of angioobliteration in the SuHx model; however, obliterated vessels did not reopen when animals with established PAH were treated with the VEGFR3 inhibitor. Part of the mechanism of vasoobliteration in the SuHx model occurs via VEGFR3. VEGFR1/VEGFR2 inhibition can be initially antiangiogenic by inducing lung vessel endothelial cell apoptosis; however, it can be subsequently angiogenic via VEGF-C and VEGF-D signaling through VEGFR3

From Monoamine Oxidase Inhibition to Antiproliferative Activity: New Biological Perspectives for Polyamine Analogs

: Monoamine oxidases (MAOs) are well-known pharmacological targets in neurological and neurodegenerative diseases. However, recent studies have revealed a new role for MAOs in certain types of cancer such as glioblastoma and prostate cancer, in which they have been found overexpressed. This finding is opening new frontiers for MAO inhibitors as potential antiproliferative agents. In light of our previous studies demonstrating how a polyamine scaffold can act as MAO inhibitor, our aim was to search for novel analogs with greater inhibitory potency for human MAOs and possibly with antiproliferative activity. A small in-house library of polyamine analogs (2-7) was selected to investigate the effect of constrained linkers between the inner amine functions of a polyamine backbone on the inhibitory potency. Compounds 4 and 5, characterized by a dianiline (4) or dianilide (5) moiety, emerged as the most potent, reversible, and mainly competitive MAO inhibitors (Ki < 1 μM). Additionally, they exhibited a high antiproliferative activity in the LN-229 human glioblastoma cell line (GI50 < 1 μM). The scaffold of compound 5 could represent a potential starting point for future development of anticancer agents endowed with MAO inhibitory activity

Ambient PM exposure and DNA methylation in tumor suppressor genes: a cross-sectional study

Exposure to ambient air particles matter (PM) has been associated with increased risk of lung cancer. Aberrant tumor suppressor gene promoter methylation has emerged as a promising biomarker for cancers, including lung cancer. Whether exposure to PM is associated with peripheral blood leukocyte (PBL) DNA methylation in tumor suppressor genes has not been evaluated. In 63 male healthy steel workers with well-characterized exposure to metal-rich particles nearby Brescia, Italy, we evaluated whether exposure to PM and metal components was associated with PBL DNA methylation in 4 tumor suppressor genes (i.e., APC, p16, p53 and RASSF1A). Blood samples were obtained on the 1st (baseline) and 4th day (post-exposure) of the same work week and DNA methylation was measured using pyrosequencing. A linear mixed model was used to examine the correlations of the exposure with promoter methylation levels. Mean promoter DNA methylation levels of APC or p16 were significantly higher in post-exposure samples compared to that in baseline samples (p-values = 0.005 for APC, and p-value = 0.006 for p16). By contrast, the mean levels of p53 or RASSF1A promoter methylation was decreased in post-exposure samples compared to that in baseline samples (p-value = 0.015 for p53; and p-value < 0.001 for RASSF1A). In post-exposure samples, APC methylation was positively associated with PM10 (β = 0.27, 95% CI: 0.13-0.40), and PM1 (β = 0.23, 95% CI: 0.09-0.38). In summary, ambient PM exposure was associated with PBL DNA methylation levels of tumor suppressor genes of APC, p16, p53 and RASSF1A, suggesting that such methylation alterations may reflect processes related to PM-induced lung carcinogenesis

Exposure to Metal-rich Particulate Matter Modifies the Expression of Candidate Micrornas in Peripheral Blood Leukocytes

Background: Altered patterns of gene expression mediate the effects of particulate matter (PM) on human health, but mechanisms through which PM modifies gene expression are largely undetermined. MicroRNAs (miRNAs) are highly conserved, noncoding small RNAs that regulate the expression of broad gene networks at the posttranscriptional level. Objectives: We evaluated the effects of exposure to PM and PM metal components on candidate miRNAs (miR-222, miR-21, and miR-146a) related with oxidative stress and inflammatory processes in 63 workers at an electric-furnace steel plant. Methods: We measured miR-222, miR-21, and miR-146a expression in blood leukocyte RNA on the first day of a workweek (baseline) and after 3 days of work (postexposure). Relative expression of miRNAs was measured by real-time polymerase chain reaction. We measured blood oxidative stress (8-hydroxyguanine) and estimated individual exposures to PM$_{1}$PM$_{1}$ (< 1 μm in aerodynamic diameter), PM$_{10}$ (< 10 μm in aerodynamic diameter), coarse PM (PM$_{10}$ minus PM$_{1}$), and PM metal components (chromium, lead, cadmium, arsenic, nickel, manganese) between the baseline and postexposure measurements. Results: Expression of miR-222 and miR-21 (using the 2$^{−ΔΔCT}$ method) was significantly increased in postexposure samples (miR-222: baseline = 0.68 ± 3.41, postexposure = 2.16 ± 2.25, p = 0.002; miR-21: baseline = 4.10 ± 3.04, postexposure = 4.66 ± 2.63, p = 0.05). In postexposure samples, miR-222 expression was positively correlated with lead exposure (β = 0.41, p = 0.02), whereas miR-21 expression was associated with blood 8-hydroxyguanine (β = 0.11, p = 0.03) but not with individual PM size fractions or metal components. Postexposure expression of miR-146a was not significantly different from baseline (baseline = 0.61 ± 2.42, postexposure = 1.90 ± 3.94, p = 0.19) but was negatively correlated with exposure to lead (β = −0.51, p = 0.011) and cadmium (β = −0.42, p = 0.04). Conclusions: Changes in miRNA expression may represent a novel mechanism mediating responses to PM and its metal components

Inhalable Metal-Rich Air Particles and Histone H3K4 Dimethylation and H3K9 Acetylation in a Cross-sectional Study of Steel Workers

Background: Epidemiology investigations have linked exposure to ambient and occupational air particulate matter (PM) with increased risk of lung cancer. PM contains carcinogenic and toxic metals, including arsenic and nickel, which have been shown in in vitro studies to induce histone modifications that activate gene expression by inducing open-chromatin states. Whether inhalation of metal components of PM induces histone modifications in human subjects is undetermined

Effects of Short-Term Exposure to Inhalable Particulate Matter on Telomere Length, Telomerase Expression, and Telomerase Methylation in Steel Workers

Shortened leukocyte telomere length (LTL) is a marker of cardiovascular risk that has been recently associated with long-term exposure to ambient particulate matter (PM). However, LTL is increased during acute inflammation and allows for rapid proliferation of inflammatory cells. Whether short-term exposure to proinflammatory exposures such as PM increases LTL has never been evaluated.We investigated the effects of acute exposure to metal-rich PM on blood LTL, as well as molecular mechanisms contributing to LTL regulation in a group of steel workers with high PM exposure.We measured LTL, as well as mRNA expression and promoter DNA methylation of the telomerase catalytic enzyme gene [human telomerase reverse transcriptase (hTERT)] in blood samples obtained from 63 steel workers on the first day of a workweek (baseline) and after 3 days of work (postexposure).LTL was significantly increased in postexposure (mean \ub1 SD, 1.43 \ub1 0.51) compared with baseline samples (1.23 \ub1 0.28, p-value < 0.001). Postexposure LTL was positively associated with PM\u2081\u2080 (\u3b2 = 0.30, p-value = 0.002 for 90th vs. 10th percentile exposure) and PM\u2081 (\u3b2 = 0.29, p-value = 0.042) exposure levels in regression models adjusting for multiple covariates. hTERT expression was lower in postexposure samples (1.31 \ub1 0.75) than at baseline (1.68 \ub1 0.86, p-value < 0.001), but the decrease in hTERT expression did not show a dose-response relationship with PM. We found no exposure-related differences in the methylation of any of the CpG sites investigated in the hTERT promoter.Short-term exposure to PM caused a rapid increase in blood LTL. The LTL increase did not appear to be mediated by PM-related changes in hTERT expression and methylation