Nitric Oxide Regulates Immune Cell Bioenergetic: A Mechanism to Understand Immunomodulatory Functions of Nitric Oxide-Releasing Anti-Inflammatory Drugs

Abstract The 2-(acetyloxy)benzoic acid 3-(nitrooxymethyl)phenyl ester (NCX-4016) is a NO-releasing derivative of aspirin. In this study, we provide evidence that NCX-4016 delivered to PMBC-derived T lymphocytes and monocytes causes a transitory inhibition of cell respiration and ≈50% reduction of cellular ATP, which translates in a time-reversible inhibition of cell proliferation and IL-2, IL-4, IL-5, and IFN-γ secretion. Exposure of lymphocytes and monocytes to aspirin, 2-(acetyloxy)benzoic acid 3-(hydroxymethyl)phenyl ester (NCX-4017), a non-NO-releasing analog of NCX-4016, and cyclooxygenase inhibitors, reduced PG formation, but has no effect on cytokine/chemokine release. In contrast, delivering NO with (z)-1-[2-(2-aminoethyl)-N-(2-ammonioethyl)amino] diazen-1-ium-1,2 diolate (DETA-NO) reproduced most of the metabolic and anti-cytokine activities of NCX-4016. Scavenging NO with hemoglobin or adding selective substrates of complex II, III, and IV of the mitochondrial respiratory chain reverses NCX-4016′ inhibitory activities. Exposure to DETA-NO and NCX-4016 enhances glucose uptake, glycolytic rate, and lactate generation in CD3/CD28-costimulated lymphocytes, while reduced citric acid cycle intermediates. These effects were not reproduced by selective and nonselective cyclooxygenase 2 inhibitors. In summary, we demonstrated that exposure of lymphocytes to NCX-4016 causes a metabolic hypoxia that inhibits lymphocyte reactivity to costimulatory molecules, providing a potential counteregulatory mechanism to control activated immune system

Regulatory T cells with multiple suppressive and potentially pro-tumor activities accumulate in human colorectal cancer

Tregs can contribute to tumor progression by suppressing antitumor immunity. Exceptionally, in human colorectal cancer (CRC), Tregs are thought to exert beneficial roles in controlling pro-tumor chronic inflammation. The goal of our study was to characterize CRC-infiltrating Tregs at multiple levels, by phenotypical, molecular and functional evaluation of Tregs from the tumor site, compared to non-tumoral mucosa and peripheral blood of CRC patients. The frequency of Tregs was higher in mucosa than in blood, and further significantly increased in tumor. Ex vivo, those Tregs suppressed the proliferation of tumor-infiltrating CD8+ and CD4+ T cells. A differential compartmentalization was detected between Helioshigh and Helioslow Treg subsets (thymus-derived versus peripherally induced): while Helioslow Tregs were enriched in both sites, only Helioshigh Tregs accumulated significantly and specifically in tumors, displayed a highly demethylated TSDR region and contained high proportions of cells expressing CD39 and OX40, markers of activation and suppression. Besides the suppression of T cells, Tregs may contribute to CRC progression also through releasing IL-17, or differentiating into Tfr cells that potentially antagonize a protective Tfh response, events that were both detected in tumor-associated Tregs. Overall, our data indicate that Treg accumulation may contribute through multiple mechanisms to CRC establishment and progression

Novel mechanisms of neuroprotective effects of Quercetin on human striatal neuroblasts

Human striatal precursor (HSP) primary cell cultures were isolated from ganglionic eminence of 9-12 week old human fetuses and extensively characterized in vitro (1). Our studies demonstrated that these cultures consists of a mixed population of neural stem cells, neuronal-restricted progenitors and striatal neurons that express and are responsive to many trophic factors, as BDNF and FGF2, and possess an adaptive response to stress conditions as nutrient deprivation and hypoxia through mechanisms involving different factors and neurotrophins (1,2). In the last decades, several in vitro and in vivo studies have provided evidence for neuroprotective effects by Quercetin, a polyphenol widely present in nature, passively absorbed in the small intestine and able to traverse the blood brain barrier (3). However, the mechanisms through which Quercetin exerts its neuroprotective effects are not fully delucidated. Our study was aimed at investigating the effects of Quercetin on HSP cells and its contribution to cell survival in nutrient deprivation condition, obtained replacing culture medium with Phosphate Buffer Saline (PBS). Quercetin treatment significantly promoted cell survival and strongly decreased apoptosis induced by nutrient deprivation condition, as evaluated by MTT assay, Trypan Blue staining and western blot analysis of cell death and proliferation markers. Moreover, since the adhesive capacities of cells are essential for cell survival, we next analysed the expression of some adhesion molecules such as Pancadherin and Focal Adhesion Kinase; our results interestingly showed that PBS exposure determined a strong decrease in all the analysed adhesion molecules, while in presence of Quercetin the expression was significantly increased. Our results add new mechanicistic insights into the comprehension of neuroprotective action of Quercetin treatment, thus suggesting possible implications in sustaining striatal neuron survival during neurodegenerative disorders, such as Huntington Disease

Cremona Beside Caregivers: una ricerca per assistere chi assiste

Stato dell’arte In Europa la fascia di popolazione over 65 rappresenta il 19% della popolazione, mentre In Italia ha già superato il 22%. Tuttavia, la distribuzione di queste persone non è omogenea sul territorio, ma presenta una maggiore presenza nelle aree rurali dove tale fascia di popolazione ha un alto rischio di esclusione sociale e fragilità (Burholt & Dobbs, 2012). Sebbene siano stati posti per molto tempo in secondo piano, i bisogni delle persone anziane nelle aree rurali sono di primario interesse, soprattutto per la diversità di richieste e di necessità che ci si aspetta di trovare in queste aree rispetto a zone con buona presenza di servizi. In questi contesti, infatti, un ruolo cruciale nell’assistenza agli anziani viene svolto dai caregiver, che si trovano largamente caricati del supporto all’invecchiamento (Sixsmith et al., 2014). Nel 2009 è stato infatti rilevato che in Italia due terzi delle cure necessarie alle persone anziane venivano assicurate da loro parenti (Troisi & Knodratowitz, 2013), anche per garantire c alle persone anziane di poter continuare a vivere nei contesti a loro cari, attraverso pratiche di “aging in place” - cioè di invecchiamento attivo nel proprio contesto comunitario di vita (Rodríguez-Rodríguez & Sánchez-González, 2016). In questo scenario, la provincia di Cremona rappresenta per sue caratteristiche territoriali e sociali un osservatorio privilegiato per studiare le necessità della popolazione anziana e di chi li assiste, anche nell’ottica di promuovere una rappresentazione propositiva e partecipe di questa fase della popolazione alla co-definizione dei servizi e dei prodotti ad essi dedicati. Date queste premesse, l’obiettivo di questo progetto di ricerca è di condurre una mappatura degli stakeholders sul territorio Cremonese, delle risorse presenti e dei bisogni scoperti, al fine di orientare attività di supporto dedicate prioritariamente al target dei caregiver. Approccio metodologico A copertura dei suddetti obiettivi si è definito un disegno di ricerca multi-fase e multi-metodo così composto: Fase 1 - analisi sistematica dei database statistici Istat, volta a descrivere le caratteristiche socio-demografiche della popolazione anziana cremonese, sia attuale che in trend storico, comparandola con quella nazionale; Fase 2 - Desk analysis per identificare i servizi sanitari, socio-sanitari e sociale attivi sul territorio a favore dell’anziano fragile che vive a casa, in termini di ruoli e i professionisti coinvolti, le collaborazioni formali e informali tra professionisti e tra Enti; regimi dietetici proposti agli anziani (i.e. in famiglia nelle RSA, nelle cooperative, nell’ADI…); Fase 3 - Interviste qualitative biografiche a stakeholders locali (i.e. rappresentanti Fondazioni, Cooperative, Comuni, RSA, associazioni di volontariato…) riconosciuti per la loro centralità nel rapporto con anziani e caregiver, al fine di approfondire le loro esperienze, l’analisi delle priorità e delle necessità presenti sul territorio. Principali risultati I dati demografici confermano la tendenza alla senilizzazione della provincia di Cremona: è la seconda provincia con la percentuale di over 65 della Regione Lombardia, dopo Pavia (Elaborazione Ires L. Morosini su dati Istat, 2019). Negli ultimi venti anni, tale percentuale è cresciuta dal 23,7% nel 2002 al 26,8% di oggi (Istat, 2021). La durata di vita attesa degli anziani cremonesi oltre i 65 anni resta inferiore a quella media lombarda: 22,1 anni nelle donne e 18,4 negli uomini, di cui la maggior parte trascorsi “senza limitazioni funzionali” (Elaborazione Ires L. Morosini su dati Istat, 2019). Tuttavia, nel 2016 il 10,8% degli ultrasessantacinquenni residenti nella Provincia di Cremona beneficiava di un’indennità di accompagnamento, il 13,8% tra le donne e il 6,9% tra gli uomini di quella fascia di età. Dalla mappatura degli stakeholders sono stati identificati 36 enti del settore sanitario, 35 del terzo settore e 7 istituzioni chiave per lo studio del fenomeno degli anziani e dei loro caregiver. Dalle prime interviste biografiche condotte emerge la necessità di dotarsi di strumenti di mappatura sistematica della fragilità senile; inoltre gli stakeholder sottolineano l’importanza di creare spazi di lavoro multidisciplinari che possano operare nelle diverse aree di ci la fragilità senile si compone (sanitaria, sociale, alimentare, economica, …). Conclusioni I primi risultati confermano il bisogno del territorio cremonese nell’ambito dell’assistenza agli anziani fragili: le attività e le risorse finora messe in campo necessitano di essere orchestrate per operare in maniera efficace e sostenibile per il territorio stesso e per le persone che lo vivono

The association between Major Depressive Disorder and premature death risk in hematologic and solid cancer: a longitudinal cohort study

Background: the aim was to verify the association between Major Depressive Disorders (MDD) and the risk of premature death in people with oncological diseases, and to collect evidence about the causality of a possible association from a longitudinal perspective.Design and Methods: it is a cohort study lasting 9 months, involving people with solid or hematologic cancers. The assessment was conducted by an ad hoc form to collect socio-demographic and clinical-oncological data, the PHQ-9 to screen MDD (cut-off ≥10) and the SF-12 to evaluate HRQoL. Relative Risk (RR) of early death between MDD exposed and not-exposed and Kaplan-Meier survival were carried out.Results: people exposed to MDD during the follow-up were 107/263 (40.7%). Among them, 36 deceased during the observation period. Overtime, having MDD and death’ occurrence showed a strong association (RR=2.15; 95% CI (1.10-4.20); χ²=5.224, p=0.0022), confirmed by Kaplan-Meier survival analysis (χ²=4.357, p=0.037). Among people who died, there was not any association between MDD, age, gender, HRQoL, cancer stage and site.Conclusions: the study confirms the association between MDD and early death in people with cancer. The absence of any association between the onset of MDD and advanced stage of cancer may suggest that it could be due to the consequences of MDD in worsening the clinical conditions related to cancer. The findings point out the relevance of MDD’ early detention among people with cancer

Place4Carers: a mixed-method study protocol for engaging family caregivers in meaningful actions for successful ageing in place

IntroductionEngaging family caregivers could be a critical asset to make the \u2018ageing-in-place\u2019 imperative a reality. This is particularly evident in rural and remote areas, where caregivers can fill the gaps that exist due to the fragmentation of the welfare system. However, there is little knowledge about the expectations that family caregivers have from healthcare services in rural and remote areas.Place4Carers (P4C) project aims to co-produce an innovative organisational model of social and healthcare services for family caregivers of older citizens living in Vallecamonica (Italy). The project is expected to facilitate ageing-in-place for older citizens, thus helping caregivers in their daily care activities.Methods and analysisP4C is a community-basedparticipatory research project featuring five work packages (WPs). WP1 consists of a survey of unmet needs of caregivers and older people receiving services in Vallecamonica. WP2 consists of a scoping literature review to map services that provide interventions of support to caregivers living in remote areas and promote engagement. WP3 organises co-creation workshops with caregivers to co-design, co-manage, and co-assess ideas and proposalsfor shaping caregiver-oriented services and organisational models. WP3 enriches the results of WP1 (survey) and WP2 (scoping literature review), and aims to co-create new ideas for intervention support with and for caregivers in relation to the objectives, features and characteristics of a new service able to address the caregivers\u2019 needs and expectations. WP4 tests the service ideas co-created in WP3 through piloting an intervention based on ideas co-created with caregivers. Finally, WP5 assesses the transferability of the intervention to other similar contexts. Ethics and dissemination. The study has been approved by the Ethics Committees of the Department of Psychology of Universit\ue0 Cattolica del Sacro Cuore and Politecnico of Milan. Results will be disseminated through peer-reviewed journals, scientific meetings and meetings with the general populatio

Ontogenetic Profile of the Expression of Thyroid Hormone Receptors in Rat and Human Corpora Cavernosa of the Penis

Introduction. In the last few years, various studies have underlined a correlation between thyroid function and male sexual function, hypothesizing a direct action of thyroid hormones on the penis. Aim. To study the spatiotemporal distribution of mRNA for the thyroid hormone nuclear receptors (TR) alpha 1, alpha 2 and beta in the penis and smooth muscle cells (SMCs) of the corpora cavernosa of rats and humans during development. Methods. We used several molecular biology techniques to study the TR expression in whole tissues or primary cultures from human and rodent penile tissues of different ages. Main Outcome Measure. We measured our data by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) amplification, Northern blot and immunohistochemistry. Results. We found that TR alpha 1 and TR alpha 2 are both expressed in the penis and in SMCs during ontogenesis without development-dependent changes. However, in the rodent model, TR beta shows an increase from 3 to 6 days post natum (dpn) to 20 dpn, remaining high in adulthood. The same expression profile was observed in humans. While the expression of TR beta is strictly regulated by development, TR alpha 1 is the principal isoform present in corpora cavernosa, suggesting its importance in SMC function. These results have been confirmed by immunohistochemistry localization in SMCs and endothelial cells of the corpora cavernosa. Conclusions. The presence of TRs in the penis provides the biological basis for the direct action of thyroid hormones on this organ. Given this evidence, physicians would be advised to investigate sexual function in men with thyroid disorders. Carosa E, Di Sante S, Rossi S, Castri A, D'Adamo F, Gravina GL, Ronchi P, Kostrouch Z, Dolci S, Lenzi A, and Jannini EA. Ontogenetic profile of the expression of thyroid hormone receptors in rat and human corpora cavernosa of the penis. J Sex Med 2010;7:1381-1390

Bulbar involvement and cognitive features in amyotrophic lateral sclerosis: a retrospective study on 347 patients

BackgroundThis study aimed at clarifying the role of bulbar involvement (BI) as a risk factor for cognitive impairment (CI) in non-demented amyotrophic lateral sclerosis (ALS) patients.MethodsData on N = 347 patients were retrospectively collected. Cognition was assessed via the Edinburgh Cognitive and Behavioral ALS Screen (ECAS). On the basis of clinical records and ALS Functional Rating Scale-Revised (ALSFRS-R) scores, BI was characterized as follows: (1) BI at onset—from medical history; (2) BI at testing (an ALSFRS-R-Bulbar score ≤11); (3) dysarthria (a score ≤3 on item 1 of the ALSFRS-R); (4) severity of BI (the total score on the ALSFRS-R-Bulbar); and (5) progression rate of BI (computed as 12-ALSFRS-R-Bulbar/disease duration in months). Logistic regressions were run to predict a below- vs. above-cutoff performance on each ECAS measure based on BI-related features while accounting for sex, disease duration, severity and progression rate of respiratory and spinal involvement and ECAS response modality.ResultsNo predictors yielded significance either on the ECAS-Total and -ALS-non-specific or on ECAS-Language/-Fluency or -Visuospatial subscales. BI at testing predicted a higher probability of an abnormal performance on the ECAS-ALS-specific (p = 0.035) and ECAS-Executive Functioning (p = 0.018). Lower ALSFRS-R-Bulbar scores were associated with a defective performance on the ECAS-Memory (p = 0.025). No other BI-related features affected other ECAS performances.DiscussionIn ALS, the occurrence of BI itself, while neither its specific features nor its presence at onset, might selectively represent a risk factor for executive impairment, whilst its severity might be associated with memory deficits

The impact of upper motor neuron involvement on clinical features, disease progression and prognosis in amyotrophic lateral sclerosis

ObjectivesIn amyotrophic lateral sclerosis (ALS) both upper (UMNs) and lower motor neurons (LMNs) are involved in the process of neurodegeneration, accounting for the great disease heterogeneity. We evaluated the associations of the burden of UMN impairment, assessed through the Penn Upper Motor Neuron Score (PUMNS), with demographic and clinical features of ALS patients to define the independent role of UMN involvement in generating disease heterogeneity, predicting disease progression and prognosis.MethodsWe collected the following clinical parameters on a cohort of 875 ALS patients: age and site of onset, survival, MRC scale, lower motor neuron score (LMNS), PUMNS, ALSFRS-R, change in ALSFRS-R over time (DFS), MITOS and King’s staging systems (KSS). Transcranial magnetic stimulation was performed on a subgroup of patients and central motor conduction time (CMCT) and cortical silent period (CSP) were calculated.ResultsWe observed that patients with an earlier age at onset and bulbar onset had higher PUMNS values. Higher values were also associated to lower ALSFRS-R and to higher DFS scores, as well as to higher MITOS and KSS, indicating that a greater UMN burden correlates with disease severity. Conversely, we did not appreciate any association between UMN involvement and survival or markers of LMN impairment. Moreover, PUMNS values showed a positive association with CMCT and a negative one with CSP values.InterpretationOur results suggest that the burden of UMN pathology, assessed through PUMNS, has an important independent role in defining clinical characteristics, functional disability, disease progression and prognosis in ALS patients. We also support the role of TMS in defining severity of UMN involvement