High Data Output and Automated 3D Correlative Light–Electron Microscopy Method

Correlative light/electron microscopy (CLEM) allows the simultaneous observation of a given subcellular structure by fluorescence light microscopy (FLM) and electron microscopy. The use of this approach is becoming increasingly frequent in cell biology. In this study, we report on a new high data output CLEM method based on the use of cryosections. We successfully applied the method to analyze the structure of rough and smooth Russell bodies used as model systems. The major advantages of our method are (i) the possibility to correlate several hundreds of events at the same time, (ii) the possibility to perform three-dimensional (3D) correlation, (iii) the possibility to immunolabel both endogenous and recombinantly expressed proteins at the same time and (iv) the possibility to combine the high data analysis capability of FLM with the high precision–accuracy of transmission electron microscopy in a CLEM hybrid morphometry analysis. We have identified and optimized critical steps in sample preparation, defined routines for sample analysis and retracing of regions of interest, developed software for semi/fully automatic 3D reconstruction and defined preliminary conditions for an hybrid light/electron microscopy morphometry approach

Characterization of a repertoire of tomato fruit genetic variants in the San Marzano genetic background

[EN] San Marzano (SM) is a worldwide famous tomato Italian traditional landrace characterized by elongated fruits with a dual-purpose use in the fresh and processing market. A repertoire of mutations affecting the fruit and of interest for commercial breeding were introduced into the SM genetic background following backcross schemes. The lines generated included 13 genotypes each carrying a single mutation in genes controlling a) the content of all pigments (hp-1, hp-2, pd), b) of carotenoids (r, t, at, B, B_mo(B)), c) of chlorophyll (gf), d) of flavonoids (y) or e) the ripening process (Nr, rin, Gr). Five lines carrying a combination of two mutations were also included. Analysis of SNP polymorphisms showed that the genetic distance of the lines from the recurrent parent was very variable and not well predicted by the number of backcrosses because it was also a function of the dissimilarity of the donor parent. All the genotypes, together with an SM control, were grown in two consecutive years and characterized for vegetative, reproductive and fruit quality traits. Overall, the studied lines reproduced the SM typical phenotypes, but several differences also emerged as both possible negative or advantageous pleiotropic traits for fresh or processing uses and peeling. High pigment mutations confirmed the negative pleiotropic effects on plant fertility and fruit development described earlier and also negatively affected fruit post-harvest life. These latter defects were also reported in the carotenoid mutant tangerine. In contrast, absence of peel pigmentation in the y mutant was associated with positive postharvest properties as those fruit presented higher resistance to wrinkling and dehydration. Delayed ripening mutants showed positive post-harvest phenotypes, as expected. In conclusion, the study of the present repertoire of fruit variations in an elongated tomato genotype represents a contribution to expand the study of fruit physiology to unusual fruit types and to breed innovative tomato lines with valuable nutritional and technological properties.This work was supported by the Latium Region FILAS project "MIGLIORA", by the Italian Ministry of Agriculture (MiPAAF) under the AGROENER project (D.D. n. 26329, 1 april 2016) - http://agroener.crea.gov.it/and by the European Commission through-H2020 SFS-7a-2014 TRADITOM (634561).Dono, G.; Picarella, ME.; Pons Puig, C.; Santangelo, E.; Monforte Gilabert, AJ.; Granell Richart, A.; Mazzucato, A. (2020). Characterization of a repertoire of tomato fruit genetic variants in the San Marzano genetic background. Scientia horticulturae (Online). 261:1-10. https://doi.org/10.1016/j.scienta.2019.108927S11026

A machine-learning based bio-psycho-social model for the prediction of non-obstructive and obstructive coronary artery disease

Background: Mechanisms of myocardial ischemia in obstructive and non-obstructive coronary artery disease (CAD), and the interplay between clinical, functional, biological and psycho-social features, are still far to be fully elucidated. Objectives: To develop a machine-learning (ML) model for the supervised prediction of obstructive versus non-obstructive CAD. Methods: From the EVA study, we analysed adults hospitalized for IHD undergoing conventional coronary angiography (CCA). Non-obstructive CAD was defined by a stenosis < 50% in one or more vessels. Baseline clinical and psycho-socio-cultural characteristics were used for computing a Rockwood and Mitnitski frailty index, and a gender score according to GENESIS-PRAXY methodology. Serum concentration of inflammatory cytokines was measured with a multiplex flow cytometry assay. Through an XGBoost classifier combined with an explainable artificial intelligence tool (SHAP), we identified the most influential features in discriminating obstructive versus non-obstructive CAD. Results: Among the overall EVA cohort (n = 509), 311 individuals (mean age 67 ± 11 years, 38% females; 67% obstructive CAD) with complete data were analysed. The ML-based model (83% accuracy and 87% precision) showed that while obstructive CAD was associated with higher frailty index, older age and a cytokine signature characterized by IL-1β, IL-12p70 and IL-33, non-obstructive CAD was associated with a higher gender score (i.e., social characteristics traditionally ascribed to women) and with a cytokine signature characterized by IL-18, IL-8, IL-23. Conclusions: Integrating clinical, biological, and psycho-social features, we have optimized a sex- and gender-unbiased model that discriminates obstructive and non-obstructive CAD. Further mechanistic studies will shed light on the biological plausibility of these associations. Clinical trial registration: NCT02737982

Multi-messenger observations of a binary neutron star merger

On 2017 August 17 a binary neutron star coalescence candidate (later designated GW170817) with merger time 12:41:04 UTC was observed through gravitational waves by the Advanced LIGO and Advanced Virgo detectors. The Fermi Gamma-ray Burst Monitor independently detected a gamma-ray burst (GRB 170817A) with a time delay of ~1.7 s with respect to the merger time. From the gravitational-wave signal, the source was initially localized to a sky region of 31 deg2 at a luminosity distance of 40+8-8 Mpc and with component masses consistent with neutron stars. The component masses were later measured to be in the range 0.86 to 2.26 Mo. An extensive observing campaign was launched across the electromagnetic spectrum leading to the discovery of a bright optical transient (SSS17a, now with the IAU identification of AT 2017gfo) in NGC 4993 (at ~40 Mpc) less than 11 hours after the merger by the One- Meter, Two Hemisphere (1M2H) team using the 1 m Swope Telescope. The optical transient was independently detected by multiple teams within an hour. Subsequent observations targeted the object and its environment. Early ultraviolet observations revealed a blue transient that faded within 48 hours. Optical and infrared observations showed a redward evolution over ~10 days. Following early non-detections, X-ray and radio emission were discovered at the transient’s position ~9 and ~16 days, respectively, after the merger. Both the X-ray and radio emission likely arise from a physical process that is distinct from the one that generates the UV/optical/near-infrared emission. No ultra-high-energy gamma-rays and no neutrino candidates consistent with the source were found in follow-up searches. These observations support the hypothesis that GW170817 was produced by the merger of two neutron stars in NGC4993 followed by a short gamma-ray burst (GRB 170817A) and a kilonova/macronova powered by the radioactive decay of r-process nuclei synthesized in the ejecta

The Sex-Specific Detrimental Effect of Diabetes and Gender-Related Factors on Pre-admission Medication Adherence Among Patients Hospitalized for Ischemic Heart Disease: Insights From EVA Study

Background: Sex and gender-related factors have been under-investigated as relevant determinants of health outcomes across non-communicable chronic diseases. Poor medication adherence results in adverse clinical outcomes and sex differences have been reported among patients at high cardiovascular risk, such as diabetics. The effect of diabetes and gender-related factors on medication adherence among women and men at high risk for ischemic heart disease (IHD) has not yet been fully investigated.Aim: To explore the role of sex, gender-related factors, and diabetes in pre-admission medication adherence among patients hospitalized for IHD.Materials and Methods: Data were obtained from the Endocrine Vascular disease Approach (EVA) (ClinicalTrials.gov Identifier: NCT02737982), a prospective cohort of patients admitted for IHD. We selected patients with baseline information regarding the presence of diabetes, cardiovascular risk factors, and gender-related variables (i.e., gender identity, gender role, gender relations, institutionalized gender). Our primary outcome was the proportion of pre-admission medication adherence defined through a self-reported questionnaire. We performed a sex-stratified analysis of clinical and gender-related factors associated with pre-admission medication adherence.Results: Two-hundred eighty patients admitted for IHD (35% women, mean age 70), were included. Around one-fourth of the patients were low-adherent to therapy before hospitalization, regardless of sex. Low-adherent patients were more likely diabetic (40%) and employed (40%). Sex-stratified analysis showed that low-adherent men were more likely to be employed (58 vs. 33%) and not primary earners (73 vs. 54%), with more masculine traits of personality, as compared with medium-high adherent men. Interestingly, women reporting medication low-adherence were similar for clinical and gender-related factors to those with medium-high adherence, except for diabetes (42 vs. 20%, p = 0.004). In a multivariate adjusted model only employed status was associated with poor medication adherence (OR 0.55, 95%CI 0.31–0.97). However, in the sex-stratified analysis, diabetes was independently associated with medication adherence only in women (OR 0.36; 95%CI 0.13–0.96), whereas a higher masculine BSRI was the only factor associated with medication adherence in men (OR 0.59, 95%CI 0.35–0.99).Conclusion: Pre-admission medication adherence is common in patients hospitalized for IHD, regardless of sex. However, patient-related factors such as diabetes, employment, and personality traits are associated with adherence in a sex-specific manner

Global urban environmental change drives adaptation in white clover

Urbanization transforms environments in ways that alter biological evolution. We examined whether urban environmental change drives parallel evolution by sampling 110,019 white clover plants from 6169 populations in 160 cities globally. Plants were assayed for a Mendelian antiherbivore defense that also affects tolerance to abiotic stressors. Urban-rural gradients were associated with the evolution of clines in defense in 47% of cities throughout the world. Variation in the strength of clines was explained by environmental changes in drought stress and vegetation cover that varied among cities. Sequencing 2074 genomes from 26 cities revealed that the evolution of urban-rural clines was best explained by adaptive evolution, but the degree of parallel adaptation varied among cities. Our results demonstrate that urbanization leads to adaptation at a global scale

Petrolio: risorsa affidabile o detrattore territoriale diffuso? Accezione e gestione del rischio nel governo del territorio della Basilicata

Nel 2013 le royalty derivanti alla regione Basilicata dall’estrazione di petrolio nella provincia di Potenza raggiungono il valore più elevato, oltre 400 mln di Euro. Intanto, con un calo di 39.119 residenti negli ultimi 25 anni (1991-2016), pari a - 6,4%, prosegue il declino demografico della regione, in atto ormai dal decennio ’61-’71. Una regione che è ricca di petrolio, risorsa da cui a livello globale dipendono, nel bene e nel male, le sorti di molte economie, ma che, ciò nonostante, rimane in una grave crisi sociale ed economica, come la vicenda demografica sottolinea. In particolare nella provincia di Potenza, nella quale l’occupazione diretta e indotta avrebbero potuto risentire positivamente del ciclo di estrazione, il calo demografico è stato nello stesso periodo perfino più marcato (- 7,5%, pari a 29.998 residenti) rispetto al dato medio regionale. Naturalmente, non può bastare il petrolio ad impedire il declino, o almeno così non dovrebbe essere; a meno di non ritenere che sul petrolio – bene economico globale per eccellenza – possano reggersi un’intera economia e un’intera cultura sociale e del lavoro. Ciò nonostante le dinamiche demografiche suggeriscono alcuni interrogativi: che tipo di risorsa è il petrolio? Come e cosa induce in termini di sviluppo sui territori da cui viene estratto? E che ruolo gioca rispetto agli altri fattori di sviluppo? Nel caso della Basilicata e in particolare della provincia di Potenza gli altri fattori, già oggi e ancor più in futuro, potrebbero essere quelle risorse tipicamente non fungibili come i beni naturali, ambientali e paesaggistici, le risorse storico-culturali, agroturistiche, agroalimentari di qualità, nonché turistiche e per il tempo libero, che sono espressione locale e propria di questi territori. Rispetto ad essi il ciclo di estrazione del petrolio può rappresentare una risorsa compatibile oppure un detrattore territoriale. Una delle questioni cruciali, dunque, è come far convivere e come gestire i rischi (ambientali ma non solo) potenzialmente derivanti dal petrolio con le opportunità che esso stesso favorisce e, ancor più, con le prospettive di sviluppo sostenibile di più lungo periodo che, per definizione, solo le risorse locali sopra indicate dovrebbero poter costruire, e che il petrolio, invece, rischia di inibire. I redditi da occupazione (pochi), e le royalty (oggi consistenti, ma nel tempo variabili) che ne derivano, infatti, possono alimentare la dipendenza economico-finanziaria e imprenditoriale del sistema produttivo locale dal petrolio; così come tutto il ciclo produttivo e la stessa attrezzatura industriale e funzionale che ne permette il funzionamento possono agire come un potenziale detrattore diffuso nella valorizzazione delle altre opportunità territoriali e socio-economiche della regione. Questo è il tema di lavoro del contributo che si propone: verificare in che termini si pone l’alternativa tra risorsa affidabile o detrattore territoriale diffuso in ordine alla “risorsa petrolio”

Il petrolio in Basilicata, affidabilità e rischi di una risorsa globale nel governo del territorio

ABSTRACT Nel 2013 le royalty derivanti alla regione Basilicata dall’estrazione di petrolio nella provincia di Potenza raggiungono il valore più elevato, oltre 400 mln di Euro. Intanto, con un calo di 39.119 residenti negli ultimi 25 anni (1991-2016), pari a - 6,4%, prosegue il declino demografico della regione, in atto ormai dal decennio ’61-’71. Una regione che è ricca di petrolio, risorsa da cui a livello globale dipendono, nel bene e nel male, le sorti di molte economie, ma che, ciò nonostante, rimane in una grave crisi sociale ed economica, come la vicenda demografica sottolinea. In particolare nella provincia di Potenza, nella quale l’occupazione diretta e indotta avrebbero potuto risentire positivamente del ciclo di estrazione, il calo demografico è stato nello stesso periodo perfino più marcato (- 7,5%, pari a 29.998 residenti) rispetto al dato medio regionale. Naturalmente, non può bastare il petrolio ad impedire il declino, o almeno così non dovrebbe essere; a meno di non ritenere che sul petrolio – bene economico globale per eccellenza – possano reggersi un’intera economia e un’intera cultura sociale e del lavoro. Ciò nonostante le dinamiche demografiche suggeriscono alcuni interrogativi: che tipo di risorsa è il petrolio? Come e cosa induce in termini di sviluppo sui territori da cui viene estratto? E che ruolo gioca rispetto agli altri fattori di sviluppo? Nel caso della Basilicata e in particolare della provincia di Potenza gli altri fattori, già oggi e ancor più in futuro, potrebbero essere quelle risorse tipicamente non fungibili come i beni naturali, ambientali e paesaggistici, le risorse storico-culturali, agroturistiche, agroalimentari di qualità, nonché turistiche e per il tempo libero, che sono espressione locale e propria di questi territori. Rispetto ad essi il ciclo di estrazione del petrolio può rappresentare una risorsa compatibile oppure un detrattore territoriale. Una delle questioni cruciali, dunque, è come far convivere e come gestire i rischi (ambientali ma non solo) potenzialmente derivanti dal petrolio con le opportunità che esso stesso favorisce e, ancor più, con le prospettive di sviluppo sostenibile di più lungo periodo che, per definizione, solo le risorse locali sopra indicate dovrebbero poter costruire, e che il petrolio, invece, rischia di inibire. I redditi da occupazione (pochi), e le royalty (oggi consistenti, ma nel tempo variabili) che ne derivano, infatti, possono alimentare la dipendenza economico-finanziaria e imprenditoriale del sistema produttivo locale dal petrolio; così come tutto il ciclo produttivo e la stessa attrezzatura industriale e funzionale che ne permette il funzionamento possono agire come un potenziale detrattore diffuso nella valorizzazione delle altre opportunità territoriali e socio-economiche della regione. Questo è il tema di lavoro del contributo che si propone: verificare in che termini si pone l’alternativa tra risorsa affidabile o detrattore territoriale diffuso in ordine alla “risorsa petrolio”

Competing land uses and sustainable development: regional planning and natural resources uses in some vulnerable areas of South of Italy

The paper focuses on the competitive land uses generated, on one side, by the advantages related to the exploitation of endogenous energy resources, and, on the other side, by the aim of protection of natural and anthropic ecosystems – dealing with the increasing awareness of the opportunities it offers in terms of economic profit and “long lasting development” - according to two points of view: - the wide range of conflicts, impacts and risks related to both oil and renewable energy production processes; - the capacity of the public decision-making system and of the spatial planning to trigger “autonomous” and sustainable development paths. In particular, the paper focuses on the conflicts caused by the growing spread of offshore wind power plants (because of the hard management of their impacts on coastal land zones) and on the impacts of oil exploitation activities on the wide “inland” and “fragile” areas (where oil production process starts) considering its effects on coastal areas (where oil is delivered and refined). The areas under investigation are the inland areas of Basilicata included in the extraction basin of the “Viggiano Oli” Center, in the Province of Potenza, from which originates an oil production cycle that reaches the coastal areas of Puglia (in particular that of the Province of Taranto). In addition, the coastal areas of Puglia will be considered as regards to the impacts of the offshore wind farms. Some final considerations concern the role of the land use and maritime spatial planning in steering a “resilience” strategy for the areas involved in energy supply activities, considering the analysis of the “multidimensional” risks (environmental, economic and social) triggered by the hard economy of oil, or by the exploitation of renewable energy resources, such as the wind power