In vitro folliculogenesis in mammalian models: a computational biology study

In vitro folliculogenesis (ivF) has been proposed as an emerging technology to support follicle growth and oocyte development. It holds a great deal of attraction from preserving human fertility to improving animal reproductive biotechnology. Despite the mice model, where live offspring have been achieved,in medium-sized mammals, ivF has not been validated yet. Thus, the employment of a network theory approach has been proposed for interpreting the large amount of ivF information collected to date in different mammalian models in order to identify the controllers of the in vitro system. The WoS-derived data generated a scale-free network, easily navigable including 641 nodes and 2089 links. A limited number of controllers (7.2%) are responsible for network robustness by preserving it against random damage. The network nodes were stratified in a coherent biological manner on three layers: the input was composed of systemic hormones and somatic- oocyte paracrine factors; the intermediate one recognized mainly key signaling molecules such as PI3K, KL, JAK-STAT, SMAD4, and cAMP; and the output layer molecules were related to functional ivF endpoints such as the FSH receptor and steroidogenesis. Notably, the phenotypes of knock-out mice previously developed for hub.BN indirectly corroborate their biological relevance in early folliculogenesis. Finally, taking advantage of the STRING analysis approach, further controllers belonging to the metabolic axis backbone were identified, such as mTOR/FOXO, FOXO3/SIRT1, and VEGF, which have been poorly considered in ivF to date. Overall, this in silico study identifies new metabolic sensor molecules controlling ivF serving as a basis for designing innovative diagnostic and treatment methods to preserve female fertility

Equine chorionic gonadotropin as an effective fsh replacement for in vitro ovine follicle and oocyte development

The use of assisted reproductive technologies (ART) still requires strategies through which to maximize individual fertility chances. In vitro folliculogenesis (ivF) may represent a valid option to convey the large source of immature oocytes in ART. Several efforts have been made to set up ivF cultural protocols in medium-sized mammals, starting with the identification of the most suitable gonadotropic stimulus. In this study, Equine Chorionic Gonadotropin (eCG) is proposed as an alternative to Follicle Stimulating Hormone (FSH) based on its long superovulation use, trans-species validation, long half-life, and low costs. The use of 3D ivF on single-ovine preantral (PA) follicles allowed us to compare the hormonal effects and to validate their influence under two different cultural conditions. The use of eCG helped to stimulate the in vitro growth of ovine PA follicles by maximizing its influence under FBS-free medium. Higher performance of follicular growth, antrum formation, steroidogenic activity and gap junction marker expression were recorded. In addition, eCG, promoted a positive effect on the germinal compartment, leading to a higher incidence of meiotic competent oocytes. These findings should help to widen the use of eCG to ivF as a valid and largely available hormonal support enabling a synchronized in vitro follicle and oocyte development

Effect of thermal/pressure processing and simulated human digestion on the immunoreactivity of extractable peanut allergens

Peanut allergy is one of the most widespread types of food allergies especially affecting developed countries. To reduce the risk of triggering allergic reactions, several technological strategies have been devised to modify or remove allergens from foods. Herein we investigated the combination of high temperature and pressure on the modulation of peanuts immunoreactivity after simulated gastro-duodenal digestion. Extractable proteins of raw and autoclaved peanuts were separated on SDS-PAGE and immunogenicity was assessed by ELISA and Western Blot analyses. Proteins surviving the heat treatment and reacting towards allergic patients' sera were analysed and attributed to Ara h 3 and Ara h 1 proteins by untargeted LC-high resolution-MS/MS. A progressive reduction in the intensity of the major allergen proteins was also highlighted in the protein fraction extracted from autoclaved peanuts, with a total disappearance of the high molecular allergens when samples were preliminary exposed to 2 h hydration although the lower molecular weight fraction was not investigated in the present work. Furthermore, raw and processed peanuts underwent simulated digestion experiments and the IgE binding was assessed by using allergic patients' sera. The persistence of an immunoreactive band was displayed around 20 kDa. In conclusion, the synergistic effects of heat and pressure played a pivotal role in the disappearance of the major peanut allergens also contributing to the significant alteration of the final immunoreactivity. In addition, the surviving of allergenic determinants in peanuts after gastrointestinal breakdown provides more insights on the fate of allergenic proteins after autoclaving treatments

“Per un Futuro di tutto Rispetto”: Sviluppo sostenibile e abitudini alimentari degli studenti toscani

membri delle Nazioni Unite, è costituita da 17 Obiettivi per lo Sviluppo Sostenibile. Essi prendono in considerazione le tre dimensioni principali dello stesso: economica, sociale ed ecologica. Sono questi alcuni importanti temi, insieme ad aspetti riguardanti le abitudini alimentari e gli stili di vita, affrontati dal progetto “sCOOL FOOD- Per un Futuro di tutto Rispetto”, finanziato dalla Fondazione Monte dei Paschi di Siena e realizzato in alcune scuole della Toscana. Scopo del nostro studio è stato quello di valutare se tale intervento, basato su attività didattica frontale, laboratoriale ed esperienziale abbia determinato sia un aumento delle conoscenze relative alla sostenibilità, che un miglioramento delle abitudini alimentari. Metodi: è stato somministrato agli studenti di scuole primarie del primo ciclo (1°-2°-3°-4°-5° elementare e 1°-2°-3° media) delle province di Siena, Arezzo e Grosseto un questionario sia all’inizio dell’anno scolastico 2021-2022 che al suo termine. In tal modo sono state raccolte informazioni riguardanti aspetti sociodemografici, le abitudini alimentari, gli stili di vita e la conoscenza sulla sostenibilità degli studenti. È stata analizzata la differenza delle risposte tra i due momenti di rilievo. Per l’analisi è stato utilizzato STATA 17. Risultati: alla prima rilevazione hanno aderito 1.592 studenti; di questi solo 520 hanno partecipato anche al questionario della seconda rilevazione. Variazioni significative tra il pre e il post intervento, si sono avute nell’aumento del consumo di frutta sia a colazione (p<0,01) che a metà mattina (p<0,01), che nella riduzione del consumo di merendine (p<0,01), panini (p<0,05), cornetti (p<0,01) e biscotti (p<0,05) a metà mattina. Per quanto riguarda la sostenibilità si è assistito ad un andamento nel complesso positivo nelle risposte corrette con variazioni statisticamente significative tra la prima e la seconda rilevazione in sette di esse: “ascoltare bene” (p<0,01), “lavorare bene insieme” (p<0,01), “regole e convenzioni comuni” (p<0,05), “l’importanza di parlare di spreco alimentare” (p<0,01), “i sensi utilizzati per degustare” (p<0,05), “la presenza di alberi in città” (p<0,01) e “eventuale possibilità di giocare con il cibo” (p<0,01). Conclusioni: Il miglioramento nelle modalità di assunzione di alcuni cibi e l’aumento delle conoscenze in alcune domande nell’ambito della sostenibilità emersi da questo studio sono a sostegno dell’efficacia di questo progetto sia nel migliorare le abitudini alimentari, che nel promuovere nei giovani comportamenti orientati alla sostenibilità, sotto il profilo economico, sociale e ambientale

Rational Discovery of Antiviral Whey Protein-Derived Small Peptides Targeting the SARS-CoV-2 Main Protease

In the present work, and for the first time, three whey protein-derived peptides (IAEK, IPAVF, MHI), endowed with ACE inhibitory activity, were examined for their antiviral activity against the SARS-CoV-2 3C-like protease (3CLpro) and Human Rhinovirus 3C protease (3Cpro) by employing molecular docking. Computational studies showed reliable binding poses within 3CLpro for the three investigated small peptides, considering docking scores as well as the binding free energy values. Validation by in vitro experiments confirmed these results. In particular, IPAVF exhibited the highest inhibitory activity by returning an IC50 equal to 1.21 μM; it was followed by IAEK, which registered an IC50 of 154.40 μM, whereas MHI was less active with an IC50 equal to 2700.62 μM. On the other hand, none of the assayed peptides registered inhibitory activity against 3Cpro. Based on these results, the herein presented small peptides are introduced as promising molecules to be exploited in the development of “target-specific antiviral” agents against SARS-CoV-2

Extrapleural pneumonectomies for pleural mesothelioma

Introduction: Malignant pleural mesothelioma (MPM) is a fatal malignancy for which there is no definitive cure. The most effective multimodality treatment in prolonging survival is still matter of debate. Surgery remains one of the cornerstones in the multimodality therapy for MPM. Extra-pleural pneumonectomy and pleurectomy/decortication are the two main curative-intent procedures; however, the superiority of one technique over the other is still debated. This review aims to assess short- and long-term results of extrapleural pneumonectomy for MPM.Areas covered: This article focuses on the role of extrapleural pneumonectomy in MPM. A systematic review was performed by using electronic databases to identify studies that included patients treated by this procedure for MPM. Endpoints included overall survival, disease-free survival, recurrence rate, perioperative mortality, and morbidity.Expert commentary: This paper offers an overview of the results that are currently obtained in patients undergoing extrapleural pneumonectomy for MPM. The benefit of surgical treatments in MPM is still debated and its primary goal should be the achievement of a macroscopic complete resection. Several alternative multimodality protocols exist, with specific advantages and drawbacks; therefore, individualization of care for each patient is fundamental. The complexity of the surgical treatment mandates that patients be referred to specialized centers

Distribution of heavy metals and polycyclic aromatic hydrocarbons in holm oak plant-soil system evaluated along urbanization gradients

Heavy metals (HMs) and polycyclic aromatic hydrocarbons (PAHs) were analysed in topsoil and in Quercus ilex L. leaves from holm oak woodlands located along urbanization gradients (urban, periurban and extraurban sites) in two Italian regions (Campania and Tuscany). In each sampling site, the metal bioavailability factors (MBFs) and the pollutant bioaccumulation factors (BAFs) were calculated to estimate the fraction of each total metal concentration in soil potentially available to root uptake and to know the fate of both HMs and PAHs in the plant-soil system. In general, the results indicated a low atmospheric deposition of pollutants and, in some cases, leaves and soils accumulated higher HM and PAH concentrations in the most urbanized areas. Correlation analyses showed that in each sampling site topsoil and leaves were exposed to the same atmospheric inputs of HMs and PAHs, although to a different extent. Notwithstanding the MBFs and BAFs differed between the two regions for the most HMs, they did not show a clear pattern in relation to the urbanization gradient. However, some information could be deduced by these ratios: the high BAF calculated for Mn shows that the foliar uptake plays an important role in accumulation of this metal. Moreover, for PAHs the different values of BAFs among low and high molecular weight compounds suggest their different fate, the first accumulated in leaves by stomata and the latter preferentially deposited on topsoi