DESIGNING AND TESTING A MOBILE APP TO FIGHT CHILD, EARLY, AND FORCED MARRIAGE IN DEVELOPING COUNTRIES

Child marriage is still a severe issue in developing countries. Among the strategies that work most to fight it, is empowering girls with information combined with the education of parents and community. As smartphones are more accessible year after year in developing countries, we want to investigate how a mobile app could effectively fight child marriage and which characteristics such an app should have. The research was organized into three main phases. The first phase was focused on 4 expert semi-structured interviews to understand if an app could be a good solution to help in fighting child marriage. The second and third phases were based on a case study with young girls in the Democratic Republic of the Congo. The second phase had focus groups with 26 girls to test and improve the requi-sites of the app, and the third phase had a questionnaire that 14 girls had to fill out after trying the app. The study shows that girls in developing countries have access to smartphones, and most can use the developed app satisfactorily and that it is worth continuing to study this problem as an app could be a new instrument to use alongside traditional tools

Simple Detection of Unstained Live Senescent Cells with Imaging Flow Cytometry

Cellular senescence is a hallmark of aging and a promising target for therapeutic approaches. The identification of senescent cells requires multiple biomarkers and complex experimental procedures, resulting in increased variability and reduced sensitivity. Here, we propose a simple and broadly applicable imaging flow cytometry (IFC) method. This method is based on measuring autofluorescence and morphological parameters and on applying recent artificial intelligence (AI) and machine learning (ML) tools. We show that the results of this method are superior to those obtained measuring the classical senescence marker, senescence-associated beta-galactosidase (SA-β-Gal). We provide evidence that this method has the potential for diagnostic or prognostic applications as it was able to detect senescence in cardiac pericytes isolated from the hearts of patients affected by end-stage heart failure. We additionally demonstrate that it can be used to quantify senescence “in vivo” and can be used to evaluate the effects of senolytic compounds. We conclude that this method can be used as a simple and fast senescence assay independently of the origin of the cells and the procedure to induce senescence

The role of procalcitonin in the follow-up of medullary thyroid cancer

Objective: Calcitonin (Ct) represents the most important biochemical marker of medullary thyroid cancer (MTC), but has certain limits. We analyzed the performance of procalcitonin (ProCt) in follow-up MTC patients. Methods: In this monocentric and retrospective study, we consecutively obtained ProCt and Ct values from all MTC patients that we visited during the period from April 2021 to May 2022. Patients were defined as having structural evidence of disease (29/90, 32.2%) irrespective of Ct values or, in its absence, as not evident disease (NED) if Ct was ≤10 ng/L (47/90, 52.2%), or minimal residual disease if Ct was >10 ng/L (14/90, 15.6%). Results: Ct and ProCt values were highly correlated (r = 0.883, P 0.12 ng/mL (P < 0.01, area under the curve: 0.963), with the following sensitivity, specificity, positive predictive value, and negative predictive value (NPV): 100%, 83.61%, 74.4%, and 100.0%. Conclusions: ProCt and Ct have a high correlation in MTC follow-up. ProCt may be useful as an adjunct to Ct, especially for its NPV concerning the structural disease

Coinvolgimento del colesterolo e del mitocondrio nella patogenesi della Sindrome di Rett

La Sindrome di Rett (RTT) è un disordine del neurosviluppo che colpisce quasi esclusivamente le bambine, e nel 95% dei casi è associato a mutazioni del gene MECP2, codificante per la proteina methyl CpG-binding protein 2 (Mecp2) associata al cromosoma X. Sebbene la patologia sia stata associata alla mutazione genica diversi anni fa e siano stati fatti enormi progressi scientifici, non è ancora chiaro l’esatto meccanismo patologico che dalla mutazione di MECP2 conduce alla manifestazione della malattia. Fra i diversi aspetti clinici della RTT, evidenze sempre maggiori suggeriscono come le disfunzioni metaboliche ed una condizione cronica di stress ossidativo ed infiammazione, possano svolgere un ruolo chiave nella manifestazione della Sindrome di RTT. In un nostro studio precedente abbiamo dimostrato come i fibroblasti RTT siano caratterizzati da un metabolismo del colesterolo alterato, con una forte riduzione del recettore SR-B1 per le HDL, a causa di modificazioni ossidative post-traduzionali. L’abilità delle HDL di esercitare le sue funzioni nel trasporto del colesterolo come anche nel possedere attività antiossidante ed antiinfiammatoria, potrebbe essere connessa alla presenza di alcune proteine accessorie quali Paraossonasi 1 (PON-1) e Fosfolipasi A2 associata alle lipoproteine (Lp-PLA2). Dati pregressi ottenuti dal nostro gruppo di ricerca, dimostrano come i fibroblasti RTT presentino un’alterata omeostasi redox a fianco di disfunzioni mitocondriali e difese antiossidanti più basse. Pertanto, in questo lavoro abbiamo investigato un possibile coinvolgimento di alterazioni del colesterolo e mitocondriali nella patogenesi della Sindrome di RTT, in particolare indagando: i) il ruolo del recettore SR-B1, degli enzimi PON-1 e Lp-PLA2 nella disregolazione del colesterolo nella RTT, determinando se i due enzimi potessero essere utilizzati per discriminare i pazienti affetti dalla RTT da quelli con disordini dello spettro autistico (ASD). ii) lo stato funzionale dei mitocondri nella RTT, focalizzandoci sulla morfologia e sul processo mitofagico e analizzando la loro relazione con le dinamiche mitocondriali (e.g. fusione e fissione) come anche con l’apoptosi, un pathway di morte cellulare che coinvolge i mitocondri. I nostri risultati mostrano un potenziale coinvolgimento della disregolazione del colesterolo rappresentata da una marcata riduzione del recettore SR-B1 nel cervello di topi RTT, presentando anche Lp-PLA2 come possibile nuovo biomarcatore per la RTT. Inoltre, abbiamo scoperto alterazioni nella morfologia dei mitocondri RTT, caratterizzati da un difetto della via mitofagica PINK1/Parkin, possibilmente connesso ad uno sbilanciamento tra fusione e fissione mediata da Drp1/Fis1, che a sua volta potrebbe essere legato ad una alterata apoptosi. Ciò mette perciò in luce un possibile ruolo chiave per questo affascinante organello nella patogenesi della Sindrome di RTT.Rett Syndrome (RTT) is an X-linked neurodevelopmental disorder that primarily affects females, associated in 95% of cases with loss-of-function mutations in MECP2 gene, encoding for methyl-CpG-binding protein 2 (Mecp2). Even if the pathology was ascribed to the gene mutation many years ago and enormous scientific progress have been done, the precise pathologic mechanism that lead from MECP2 mutations to RTT manifestation is still a lack. Among several clinical aspects of RTT, growing evidence suggest a possible key role exerted by metabolic dysfunctions linked to a detrimental vicious cycle of chronic OxInflammation state. In our previous works we have demonstrated that RTT fibroblasts showed an altered cholesterol metabolism, characterized by a strong reduction of the scavenger HDL receptor SR-B1 due to oxidative post-translation modifications. HDL’s ability to perform its functions in cholesterol transport as well as in exerting antioxidant and anti-inflammatory activity, may be related to the presence of some accessory proteins like Paraoxonase-1 (PON-1) and Lipoprotein associated phospholipase A2 (Lp-PLA2). We found also an impaired redox homeostasis represented by mitochondrial dysfunctions that parallels with a lower antioxidant defence activity. Therefore, in this work we investigated a possible involvement of cholesterol and mitochondrial impairment in RTT pathogenesis addressing the following topics: i) The role of SR-B1 receptor, PON-1 and Lp-PLA2 enzymes in cholesterol dysregulation in RTT and the determination whether the two enzymes might be used to discriminate RTT from Autism Spectrum Disorder (ASD) patients. ii) The functional status of mitochondria in RTT, with specific focus on morphology and mitophagy, taking into account also their relationship with mitochondrial dynamics (e.g. fusion and fission) as well as apoptosis, a cell death pathway involving also mitochondria. Our findings showed a potential involvement of cholesterol dysregulation via SR-B1 loss in RTT brain mice, presenting also Lp-PLA2 as a new possible biomarker for RTT. Moreover, we found alterations in the morphology of RTT mitochondria, which were characterized by a defect in PINK1/Parkin dependent-mitophagy possibly due to unbalanced mitochondrial fusion and Drp1/Fis1-mediated fission, which in turn could be related to an aberrant apoptosis. Therefore, our results highlighted a possible role for this fascinating organelle in RTT pathogenesis

Fighting child marriage in developing countries: design and development of a mobile application

Child marriage is still a great issue in developing countries and even if the interventions to prevent it are having results, they are not enough to eliminate the problem. Among the strategies that seem to work most to fight child marriage, there is the empowerment of girls with information combined with education of parents and community. As smartphones are more accessible year after year in developing countries, this thesis wants to investigate if a mobile app could be effective in fighting child marriage and which characteristics such an app should have. The research was organized in four phases and used design and creation and case study methodologies. Firstly, the literature was analyzed and an initial design was proposed. Secondly, expert interviews were performed to gain feedback on the proposed design, and afterwards prototype was built. Thirdly, a case study in the Democratic Republic of Congo (DRC) was performed to test the prototype, gaining insights and improvements through group interviews with 26 girls aged 15-19. Finally, a first version of the app was developed and a second phase of the case study was run in the DRC to understand if the girls were able to use the app. This phase included 14 girls of which 6 had participated in the prototype testing and used questionnaires as a data generation method. The app was built following the Principles for Digital Development. Even if this app is built based on the case study in DRC is modular and easily adaptable to other contexts as it is not content-specific. It was shown that is worth continuing to study this topic and it was defined a conceptual framework for designing learning apps for developing countries, in particular, to fight child, early, and forced marriage

Lactonase Activity and Lipoprotein-Phospholipase A2 as Possible Novel Serum Biomarkers for the Differential Diagnosis of Autism Spectrum Disorders and Rett Syndrome: Results from a Pilot Study

Rett syndrome (RTT) and autism spectrum disorders (ASDs) are not merely expression of brain dysfunction but also reflect the perturbation of physiological/metabolic homeostasis. Accordingly, both disorders appear to be associated with increased vulnerability to toxicants produced by redox imbalance, inflammation, and pollution, and impairment of systemic-detoxifying agents could play a role in the exacerbation of these detrimental processes. To check this hypothesis, the activities of two mechanistically related blood-based enzymes, paraoxonase-1 (arylesterase, paraoxonase, and lactonase), and lipoprotein-associated phospholipase A(2) (Lp-PLA(2)) were measured in the serum of 79 ASD and 95 RTT patients, and 77 controls. Lactonase and Lp-PLA(2) showed a similar trend characterized by significantly lower levels of both activities in ASD compared to controls and RTT (p < 0 001 for all pairwise comparisons). Noteworthy, receiving operator curve (ROC) analysis revealed that lactonase and, mostly, Lp-PLA(2) were able to discriminate between ASD and controls (lactonase: area under curve, AUC = 0.660; Lp-PLA(2), AUC = 0.780), and, considering only females, between ASD and RTT (lactonase, AUC = 0.714; Lp-PLA(2), AUC = 0.881). These results suggest that lactonase and, especially, Lp-PLA(2) activities might represent novel candidate biomarkers for ASD

ES2 as a novel verbascoside-derived compound in the treatment of cutaneous wound healing

Several pathologies are characterized by chronic wounds and often resistant to many of the common therapies, leading to chronic infections that can become even life-threatening for patients. For this reason, the identification of new products able to ameliorate the healing process is still an on-going research. Natural compounds have been used to improve skin conditions due to their dermo-cosmetic and therapeutic activities including anti-inflammatory, antioxidant and cell-migratory properties. Among these compounds, it has been recently demonstrated that Verbascoside, a phenyl propanoid glycoside widely used in the cosmetic field, can improve keratinocytes proliferation. Because of its high hydrophilic character, Verbascoside has a limited range of possible topical applications and the synthesis of ES2, a semi-synthetic derivative of Verbascoside was performed to bypass some of the drawback aspects of this molecule. In the present study, the wound healing properties of Verbascoside and ES2 were compared in both keratinocytes "in vitro" wound scratch and in wounded SKH1 mice. The results showed that both compounds were not cytotoxic and ES2 showed an efficient ability to promote the proliferation of human keratinocytes compared to Verbascoside. The findings were also confirmed in vivo but only at early time points (2/3 days). Taken together, these data suggest that the Verbascoside-derivative ES2 could be considered a novel and promising candidate for the topical treatment of wounds