Glutathione-mediated antioxidant response and aerobic metabolism: two crucial factors involved in determining the multi-drug resistance of high-risk neuroblastoma

Neuroblastoma, a paediatric malignant tumor, is initially sensitive to etoposide, a drug to which many patients develop chemoresistance. In order to investigate the molecular mechanisms responsible for etoposide chemoresistance, HTLA-230, a human MYCN-amplified neuroblastoma cell line, was chronically treated with etoposide at a concentration that in vitro mimics the clinically-used dose. The selected cells (HTLA-Chr) acquire multi-drug resistance (MDR), becoming less sensitive than parental cells to high doses of etoposide or doxorubicin. MDR is due to several mechanisms that together contribute to maintaining non-toxic levels of H2O2. In fact, HTLA-Chr cells, while having an efficient aerobic metabolism, are also characterized by an up-regulation of catalase activity and higher levels of reduced glutathione (GSH), a thiol antioxidant compound. The combination of such mechanisms contributes to prevent membrane lipoperoxidation and cell death. Treatment of HTLA-Chr cells with L-Buthionine-sulfoximine, an inhibitor of GSH biosynthesis, markedly reduces their tumorigenic potential that is instead enhanced by the exposure to N-Acetylcysteine, able to promote GSH synthesis.Collectively, these results demonstrate that GSH and GSH-related responses play a crucial role in the acquisition of MDR and suggest that GSH level monitoring is an efficient strategy to early identify the onset of drug resistance and to control the patient's response to therapy

The impact of the two-year COVID-19 pandemic on hospital admission and readmissions of children and adolescents because of mental health problems

PurposeThis study aimed to investigate the specific risk factors and psycho-social and clinical features of hospitalized neuropsychiatric patients during the COVID pandemic and to analyze the hospital readmission phenomenon, which, according to recent studies, increased in frequency during the first pandemic period.Patients and methodsThis observational retrospective cohort study examined 375 patients aged between 0 and 17 years who were hospitalized between 1 February 2018 and 31 March 2022 due to neuropsychiatric issues. The majority of the patients were girls: there were 265 girls compared to 110 boys (M = 13.9 years; SD 2.30 years). The total sample was divided into two groups: the pre-COVID-19 group (160 inpatients hospitalized between February 2018 and February 2020) and the COVID-19 group (215 inpatients hospitalized between March 2020 and March 2022). To explore the readmission phenomenon (second aim), we selected from the two groups of patients with at least one hospital readmission within 365 days after the first discharge. Multiple variables (sociodemographic, clinical, psychological, and related to hospitalization) were collected for each patient by reviewing their medical records.ResultsThe risk factors for mental health disorders were similar between the two groups, except for the significantly increased use of electronic devices in the COVID-19 group, increasing from 8.8% in the pre-COVID-19 group to 29.2% in the COVID-19 group. Patients suffering from eating disorders increased from 11.3% in the pre-COVID-19 group to 23.8% in the COVID-19 group. Hospital readmissions nearly increased from 16.7% in the 2-year pre-COVID-19 period to 26.2% in the 2-year COVID-19 period. A total of 75% of patients hospitalized three or more times in the last 2 years and 85.7% of the so-called “revolving door” patients (with relapse within 3 months after discharge) were identified in the COVID-19 group. However, the comparison between the two groups of patients readmitted before and during the COVID-19 pandemic did not show any differences in terms of sociodemographic and clinical characteristics.ConclusionIn conclusion, there was a significant increase in hospital readmissions, but these results suggest the need for better coordination between hospital and territorial services in managing the complexity of mental health problems related to situations arising from the COVID-19 pandemic and the necessity to implement prevention strategies and services

Candidate biomarkers from the integration of methylation and gene expression in discordant autistic sibling pairs

While the genetics of autism spectrum disorders (ASD) has been intensively studied, resulting in the identification of over 100 putative risk genes, the epigenetics of ASD has received less attention, and results have been inconsistent across studies. We aimed to investigate the contribution of DNA methylation (DNAm) to the risk of ASD and identify candidate biomarkers arising from the interaction of epigenetic mechanisms with genotype, gene expression, and cellular proportions. We performed DNAm differential analysis using whole blood samples from 75 discordant sibling pairs of the Italian Autism Network collection and estimated their cellular composition. We studied the correlation between DNAm and gene expression accounting for the potential effects of different genotypes on DNAm. We showed that the proportion of NK cells was significantly reduced in ASD siblings suggesting an imbalance in their immune system. We identified differentially methylated regions (DMRs) involved in neurogenesis and synaptic organization. Among candidate loci for ASD, we detected a DMR mapping to CLEC11A (neighboring SHANK1) where DNAm and gene expression were significantly and negatively correlated, independently from genotype effects. As reported in previous studies, we confirmed the involvement of immune functions in the pathophysiology of ASD. Notwithstanding the complexity of the disorder, suitable biomarkers such as CLEC11A and its neighbor SHANK1 can be discovered using integrative analyses even with peripheral tissues

Brain-Derived Neurotrophic Factor (BDNF) and IL-1beta in Pediatric and Adolescent' Depressive Disorders: a look towards precision psychiatry

Introduction. Depression is one of the main and most debilitating mood disorders, as for personal functioning and social costs (Masi and Brovedani, 2011). It is estimated that depression affects more than 15% of the population lifetime (Kessler et al., 2003), with percentages in pediatric age that reach about 1-2% in prepuberal age and 4-6% in adolescence (Kessler et al., 2001). Although the early-onset depression usually tends to improve over time and resolve in a part of the cases, the functional impact of the disease is more difficult to recover, with consequences that tend to take a chronic trend and finally predispose to a high rate of recurrence of disease, a high risk of psychopathology in the future, as well as increase risk of suicide attempts and completed suicides (Masi and Brovedani, 2011). Studies on neuroanatomy, post-mortem brain observations and in vivo neuroimaging suggest that the impairment of neuroplasticity in specific neuronal networks may be involved in the pathogenesis of mood disorders (Biggio, 2011). In particular, recent studies have emphasized the role of the neurotrophic factor called Brain Derived Neurotrophic Factor (BDNF) whose depletion is associated with a reduction in neurogenesis and in the volume of important brain areas including the hippocampus, the prefrontal cortex and the amygdala (Biggio, 2011). Other researches found reduced BDNF concentrations in the blood of depressed patients, which increased after undergoing treatment with psychopharmacological therapy (Brunoni et al., 2008; Sen et al., 2008; Bocchio-Chiavetto et al., 2010; Molendijk et al. al., 2014). Scientific studies also found that the neuroinflammation process could contribute to the origin of some psychiatric conditions, especially depression (Dantzer et al. 2008; Miller et al. 2009; Kim et al. 2014; Rosenblat et al. 2014). Depressed patients often present an increase levels of proinflammatory cytokines (like IL-1beta) in blood and cerebrospinal fluid (Miller et al, 2009; Kim et al. 2014), so many authors have hypothesized their role as early clinical biomarker of pathology. Objective. The aim of the present study is to measure plasma levels of BDNF and IL-1beta in subjects affected by depression in the developmental age compared to healthy peers. Secondary aims are: a) the evaluation in the depressed subjects of the trend of plasma levels before and after psychopharmacological treatment (at 3, 6 and 12 months); b) the analysis of the relationship between plasma levels of BDNF and IL-1beta and clinical psychopatological features of depression. The last objective is to verify the usefulness of BDNF and IL-1beta dosages as peripheral biomarkers of disease or of unresponsiveness to treatment, finally to improve diagnostic characterization of depressed patients and to target the treatment in pediatric age. Methods. Thirteen subjects aged 11-17,9 years, diagnosed with depressive disorder (according to DSM-IV-TR; APA, 2002), as well as matched healthy subjects (n =12), were evaluated by trained raters and provided blood for BDNF and IL-1beta measurements. The diagnostic evaluation was conducted through free talks and the administration of a panel of structured (YSR-CBCL, CDI, MASC, CGI, C-GAS, SadPerson) and projective tests. The same panel was adminstrated to healthy subjects to explore subthereshold dimensions. Depressed subjects were treated with pharmacological therapy (Selective Serotonin Reuptake Inhibitors, mood stabilizers, SGAs) and psychotherapy. The plasma levels of BDNF and IL-1beta were measured according to definite timing (basal T0, before the start of therapy / psychotherapy, after 6 and 12 months). For depressed patients, an additional measure after 3 months of therapy was performed. Analysis of BDNF and IL-1beta levels were performed using specific ELISA kits (BDNF Emax ImmunoAssay System, Promega, Madison, U.S.A and Antigenix America, Huntington Station, NY, USA) Results. The depressed subjects, compared to the healthy peers, differ in a statistically significant way for a constant familiarity for psychiatric pathology, in particular in the maternal line, a history of events experienced as traumatic, a constant presence of conflicts within the family, scholastic and relational problems, sleep disorders and alcohol/drugs use. Compared to heathy subjects at intake, plasma BDNF concentrations are lower, while IL-1beta are higher in depressed patients, even if they do not reach a statistical significance. Regarding psychodiagnostic assessment, comparing test scores between depressed and non-depressed subjects at T0, there are significant differences in CBCL, YSR, C-GAS, CGI-S, CDI (p <0.05) and SadPerson (p <0.001) scores. No significant differences were found between the gropus in the MASC scores. Statistically significant correlations were found between BDNF levels at T0 of depressed subjects and problems of attention (r=-.695, p=.026) and school skills (r= .878, p= .004). At T6, BDNF inversely correlates to affective disorders (r=-.975, p=.005) reported at YSR T12. In depressed group, considering the presence of suicidal attempts, there are statistical significative differences for anxious-depressed (Z=-2.158, p=.031) and anxiety problems (Z=-2.669, p=.008) at YSR. The same analysis performed considering the personal history of trauma, highlighted statistical significative differences for rule break (Z=-2.384, p=.017) and conduct disorders (Z=-2.037, p=.042). Conclusions. Depressed pediatric patients present lower levels of BDNF and higher levels of IL-1beta compared to healthy peers at the moment of diagnosis, although without a statistical significant difference. At intake BDNF appears to correlate to academic competences, while after 6 months of therapy higher BDNF levels orient to a better prognosis as for depression symptomatology. The evolution from suicidal ideation to SA appear to be linked to the perception of limited social support (reduced resilience) in adolescents with reduced levels of BDNF. Depressed children with a history of trauma present more externalizing symptoms, while the dimension of anxiety appears to be somehow protective from suicidal attempts. The study supports the bio-psycho-social model of DD, highlighting the importance of a balance between neurogenesis, neuroinflammation and cerebral plasticity in the delicate period of developmental age. BDNF seems to be a symptoms or disease biomarker more than a severity index, so the integration of different instruments of evaluations, both clinical and biological (multimodal methods), as well as the integration of different voices (multi-informant approach) seems to represent the “best practice” for pediatric clinical assessment of mental health in line with precision psychiatry. Further studies are needed to explore the role of BDNF and cytokines in the response of pediatric population to SSRIs, to identify biomarkers for predicting early treatment response, to produce new drug targets, to find preventive strategies and individually target therapy.Introduzione. La depressione è uno tra i principali e maggiormente debilitanti disturbi dell'umore, sia in termini di funzionamento personale che di costi sociali (Masi and Brovedani, 2011). Si stima che la depressione colpisca almeno una volta nella vita più del 15% della popolazione (Kessler et al., 2003), con percentuali in età pediatrica che raggiungono circa l’1-2% in epoca prepuberale e il 4-6% nell’adolescenza (Kessler et al., 2001). Nonostante la depressione ad esordio precoce solitamente tenda a migliorare nel tempo e a risolversi in una parte dei casi, l’impatto funzionale della malattia risulta più difficile da recuperare, con conseguenze che tendono ad assumere un andamento cronico e che infine predispongono ad un alto tasso di ricorrenza di malattia, un alto rischio di psicopatologia futura, tentativi di suicidio e suicidi completati (Masi et Brovedani,2011). Studi di neuroanatomia, osservazioni condotte sul cervello post-mortem e ricerche di neuroimaging in vivo suggeriscono che la compromissione della neuroplasticità a livello di specifiche reti neuronali possa essere coinvolta nella patogenesi dei disturbi dell’umore (Biggio, 2011). In particolare, studi recenti hanno sottolineato il ruolo di un fattore neurotrofico denominato Brain Derived Neurotrophic Factor (BDNF) la cui deplezione è associata a riduzione della neurogenesi e del volume di importanti aree cerebrali tra cui l’ippocampo, la corteccia prefontale e l’amigdala (Biggio, 2011). Altre ricerche hanno riscontrato concentrazioni ridotte di BDNF nel sangue di pazienti depressi e un successivo incremento a seguito di trattamento con terapia psicofarmacologica (Brunoni et al., 2008; Sen et al., 2008; Bocchio-Chiavetto et al., 2010; Molendijk et al., 2014). Altri studi scientifici evidenziano che anche processi di neuroinfiammazione possono contribuire all'origine di alcune patologie psichiatriche ed in particolar modo della depressione (Dantzer et al. 2008; Miller et al.2009; Kim et al. 2014; Rosenblat et al. 2014). I pazienti depressi infatti presentano spesso aumenti dei livelli di citochine proinfiammatorie (come l'IL-1beta) nel sangue e nel liquor (Miller et al, 2009; Kim et al. 2014), pertanto molti autori ipotizzano un loro ruolo come biomarker precoci di malattia. Scopo dello studio. Lo scopo dello studio è la misurazione dei livelli plasmatici di BDNF e di IL-1beta in soggetti affetti da depressione in età evolutiva rispetto ad un gruppo di soggetti sani. Obiettivi secondari sono: a) la valutazione nei soggetti depressi dell'andamento dei livelli plasmatici prima e dopo trattamento psicofarmacologico (a 3, 6 e 12 mesi); b) la valutazione della relazione tra livelli plasmatici di BDNF e IL-1beta e le caratteristiche cliniche psicopatologiche della depressione. Obiettivo finale è verificare l'utilità del dosaggio del BDNF e dell'IL-1beta, quali possibili biomarkers periferici di malattia o di mancata risposta alla terapia, per migliorare la caratterizzazione dei pazienti depressi e finalizzare il trattamento in età pediatrica. Materiali e metodi. Tredici soggetti di età compresa tra 11 e 17,9 anni, con diagnosi di disturbo depressivo (secondo il DSM-IV-TR, APA, 2002), e un parallelo gruppo di soggetti sani appaiati per sesso ed età (n = 12), sono stati valutati da personale specializzato. Sono stati prelevati inoltre campioni ematici per la misurazione di BDNF e IL-1beta. La diagnosi è stata condotta attraverso colloqui liberi e la somministrazione di test strutturati (YSR-CBCL, CDI, MASC, CGI, C-GAS, SadPerson) e proiettivi. Gli stessi test sono stati somministrati anche ai soggetti sani per esplorare la presenza di eventuali dimensioni depressive sottosoglia. I soggetti depressi sono stati trattati con terapia farmacologica (inibitori selettivi della ricaptazione della serotonina, stabilizzatori dell'umore, antipsicotici atipici) e psicoterapia. I livelli plasmatici di BDNF e IL-1beta sono stati misurati secondo tempi definiti (T0 basale, prima dell'inizio della terapia/ psicoterapia, dopo 6 e 12 mesi). Per i pazienti depressi, è stata eseguito un ulteriore dosaggio dopo 3 mesi di terapia. L'analisi dei livelli di BDNF e IL-1beta è stata eseguita utilizzando specifici kit ELISA (BDNF Emax ImmunoAssay System, Promega, Madison, U.S.A and Antigenix America, Huntington Station, NY, USA) Risultati. I soggetti depressi, rispetto ai coetanei sani, differiscono in modo statisticamente significativo per una costante familiarità per la patologia psichiatrica, in particolare in linea materna, una anamnesi positiva per traumi, una costante presenza di conflitti all'interno della famiglia, problemi scolastici e relazionali, disturbi del sonno e uso di alcol o sostanze. Rispetto ai soggetti sani, al momento del reclutamento, le concentrazioni plasmatiche di BDNF nei pazienti depressi risultano più basse, mentre quelle di IL-1beta più alte, tuttavia i risultati non raggiungono una significatività statistica. Per quanto riguarda la valutazione psicodiagnostica, confrontando i punteggi dei test tra soggetti depressi e non depressi a T0, vi sono differenze significative nei punteggi delle CBCL, YSR, C-GAS, CGI-S, CDI (p <0,05) e SadPerson (p <0,001). Nessuna differenza significativa è stata trovata tra i gruppi nei punteggi MASC. Nei soggetti con depressione è presente una correlazione statisticamente significativa tra i livelli di BDNF al T0 e problemi di attenzione (r = -. 695, p = 0,026) e abilità scolastiche (r = .878, p = 0,004). A T6, i livelli di BDNF sono inversamente correlati a disturbi affettivi (r = -. 975, p = 0,005) riportati al test YSR a 12 mesi. Nel gruppo di soggetti depressi, considerando la presenza di tentativi di suicidio, esistono differenze statisticamente significative per problemi ansioso/depressivi (Z = -2,158, p = 0,031) e di ansia (Z = -2,669, p = 0,008) riportati al YSR. La stessa analisi eseguita considerando l'anamnesi positiva per eventi traumatici, ha evidenziato differenze statisticamente significative per lo scarso rispetto delle regole (Z = -2.384, p = .017) e i disturbi del comportamento (Z = -2.037, p = .042). Conclusioni. I pazienti pediatrici depressi presentano livelli più bassi di BDNF e livelli più alti di IL-1beta rispetto ai coetanei sani al momento della diagnosi, sebbene il dato non raggiunga una differenza statisticamente significativa. Al momento del reclutamento, il livello di BDNF appare correlare con le competenze accademiche, mentre dopo 6 mesi di terapia livelli di BDNF più elevati orientano verso una prognosi migliore relativamente alla sintomatologia depressiva. L'evoluzione dall'ideazione suicidaria al tentato suicidio sembra essere collegata alla percezione di un limitato supporto sociale (ridotta resilienza) negli adolescenti con livelli ridotti di BDNF. I bambini depressi con una storia di trauma presentano sintomi più esternalizzanti, mentre la dimensione dell'ansia sembra essere in qualche modo protettiva rispetto al tentativo di suicidio. Lo studio supporta il modello bio-psico-sociale dei DD, evidenziando l'importanza di un equilibrio tra neurogenesi, neuroinfiammazione e plasticità cerebrale nel delicato periodo dell’età evolutiva. Il BDNF sembra essere un biomarcatore di sintomi e di patologia, piuttosto che un indice di gravità, pertanto l'integrazione di diversi strumenti di valutazione, sia clinici che biologici (metodi multimodali), così come l'integrazione di diverse voci (approccio multi-informant) sembrano rappresentare la "migliore pratica" per la valutazione clinica pediatrica della salute mentale in linea con la psichiatria di precisione. Ulteriori studi si rendono necessari al fine di esplorare il ruolo del BDNF e delle citochine nella risposta terapeutica agli SSRI nella popolazione pediatrica, per identificare biomarkers predittori precoci di risposta al trattamento, per la produzione di nuovi bersagli farmacologici, per trovare strategie preventive e mirare ad una terapia individualizzata nella depressione

Innovative cargo unit for vehicles transport: technical and economic considerations

A new intermodal container specifically designed to carry cars by both land and sea has been recently presented. This unit is a collapsible, 40-foot, ISO container with two decks and space for six cars. Because of its standard sizes, the unit will allow cars to be carried door-to-door (from factory to showrooms) by the same transportation means which carry containers all over the world. Cars were traditionally delivered using specialised land vehicles and vessels; such modes were not integrated and this fact led to multiple lashing and handling operations as vehicles were transferred from one mode to another. This paper analyses the benefits of car containerisation, mainly in terms of lower transportation costs and lesser space requirements for land Storage

HOSPITALISATION IN CHILD NEUROPSYCHIATRY: A CASE STUDY ALONG A FIVE-YEAR EPIDEMIOLOGICAL-CLINICAL TREND

Objective: In Italy, the number of patients admitted to child and adolescent neuropsychiatry services has almost doubled in the last 10 years. Despite this significant increase in demand, there is still a paucity of literature on mental disorders in the paediatric population. Therefore, we investigated and described the clinical and socio-demographic characteristics of a sample of young Italian inpatients with psychiatric disorders. The aim was to contribute to the jet scarce literature on this topic, while also providing useful information for the clinical-care organisation of mental health services dedicated to children and adolescents. Method: In this retrospective cohort study, data were collected from 361 hospitalised patients aged̀ 1 to 18 who had been admitted to a Child Neuropsychiatry Unit in Northern Italy, from January 2016 to December 2020. Descriptive analyses, Univariate Analysis of Variance (ANOVA), and Chi-square tests were applied. Results: During a five-year timeline, a higher admission rate for females was recorded, and the average age of inpatients was 13.4 years (SD = 3.01). Most of the admissions occurred through the Paediatric Emergency Department, and suicidal behaviour was the most frequent reason for admission. At discharge, affective disorders were the main diagnoses, which were also found to be the most frequent in patients with self-injurious behaviours. Non-suicidal self-injury, which was mainly reported as occurring in order to obtain relief from suffering, regarded 40.8% of the total sample. Almost half of the subjects reported suicidal ideation, and 21.1% attempted suicide. The mean hospitalisation length significantly decreased from 2016–2018 to 2019–2020. In general, patients with psychotic disorders had the longest stays. Conclusions: Ad hoc diagnostic–therapeutic protocols should be developed for psychiatric emergencies, and health personnel should be adequately trained to manage acute psychiatric conditions in developmental age. Primary and secondary prevention programs should be implemented to promptly recognise and treat mental health issues in this age group

Diagnostic and Therapeutic Challenges of Catatonia in an Adolescent With High Functioning Autism Spectrum Disorder: A Case Report

Catatonia is a psychomotor syndrome with specific clusters of speech, behavioral and motor features. Although potentially life-threatening, especially in its malignant form accompanied with autonomic dysregulation and medical complications, it is a treatable condition, when promptly identified. For a long time catatonia was considered a marker of schizophrenia, thus limiting the possibility of diagnosis and treatment. Due to growing awareness and studies on the subject, it is now known that catatonia can occur in the context of a number of diseases, including psychotic, affective and neurodevelopmental disorders. In recent years, there's been a renewed interest in the recognition and definition of catatonia in neurodevelopmental disorders, such as Autism Spectrum Disorder (ASD), where the differential diagnosis poses great challenges, given the considerable overlapping of signs and symptoms between the conditions. We present the case of a 15 year old boy with High Functioning ASD with a sudden onset of severe catatonic symptoms and the co-existence of psychotic symptoms, whose complex clinical course raises many questions on the differentiation and relation of said disorders