CFTR Modulation Reduces SARS-CoV-2 Infection in Human Bronchial Epithelial Cells

People with cystic fibrosis should be considered at increased risk of developing severe symptoms of COVID-19. Strikingly, a broad array of evidence shows reduced spread of SARS-CoV-2 in these subjects, suggesting a potential role for CFTR in the regulation of SARS-CoV-2 infection/replication. Here, we analyzed SARS-CoV-2 replication in wild-type and CFTR-modified human bronchial epithelial cell lines and primary cells to investigate SARS-CoV-2 infection in people with cystic fibrosis. Both immortalized and primary human bronchial epithelial cells expressing wt or F508del-CFTR along with CRISPR/Cas9 CFTR-ablated clones were infected with SARS-CoV-2 and samples were harvested before and from 24 to 72 h post-infection. CFTR function was also inhibited in wt-CFTR cells with the CFTR-specific inhibitor IOWH-032 and partially restored in F508del-CFTR cells with a combination of CFTR modulators (VX-661+VX-445). Viral load was evaluated by real-time RT-PCR in both supernatant and cell extracts, and ACE-2 expression was analyzed by both western blotting and flow cytometry. SARS-CoV-2 replication was reduced in CFTR-modified bronchial cells compared with wild-type cell lines. No major difference in ACE-2 expression was detected before infection between wild-type and CFTR-modified cells, while a higher expression in wild-type compared to CFTR-modified cells was detectable at 72 h post-infection. Furthermore, inhibition of CFTR channel function elicited significant inhibition of viral replication in cells with wt-CFTR, and correction of CFTR function in F508del-CFTR cells increased the release of SARS-CoV-2 viral particles. Our study provides evidence that CFTR expression/function is involved in the regulation of SARS-CoV-2 replication, thus providing novel insights into the role of CFTR in SARS-CoV-2 infection and the development of therapeutic strategies for COVID-19

Studio del processo di breakup in reazioni nucleari indotte da Fasci Radioattivi Leggeri

In questo lavoro, sono state studiate le reazioni (_^7)Be+ (_^208)Pb e (_^8)B+ (_^208)Pb ad energie attorno alla barriera coulombiana. (_^7)Be e (_^8)B sono isotopi instabili appartenenti alla grande classe dei fasci radioattivi, che ha acquisito un interesse sempre maggiore negli ultimi anni, soprattutto nello studio di nuovi fenomeni di struttura nucleare, di nuovi meccanismi di reazione e di nuove modalità di decadimento, in regioni della carta dei nuclidi lontane dalla valle della stabilità. Lo scopo di questo lavoro di tesi è quello di effettuare un primo tentativo di investigazione su quale processo nucleare sia il principale responsabile dell'evidente aumento della sezione d'urto di reazione del (_^7)Be e (_^8)B misurato sperimentalmente. Inoltre, tenendo in considerazione il fatto che il nucleo di (_^8)B è composto da un core di (_^7)Be+p , per questo motivo è importante estrarre dai dati sperimentali la distribuzione angolare del (_^7)Be prodotto nella reazione(_^8)B+ (_^208)Pb , nonché studiare anche la reazione (_^7)Be+ (_^208)Pb stessa ed i suoi prodotti di reazione. Pertanto, lo studio del (_^7)Be è importante per approfondire la nostra comprensione delle proprietà e della struttura del (_^8)B . Il mio lavoro di tesi è stato quello di partire da questo file di dati e svolgere una serie di operazioni di analisi dati con lo scopo di ottenere i seguenti risultati: • La sezione d'urto differenziale degli isotopi (_^3)He+ (_^4)He prodotti nella reazione (_^7)Be+ (_^208)Pb a 40.5 MeV di energia; • La sezione d'urto differenziale del (_^7)Be prodotto nella reazione (_^8)B+ (_^208)Pb ad un'energia di 50 MeV. Nella prima parte di introduzione è stato trattato il tema dei nuclei esotici, in particolare ci siamo interrogati se le basse energie di legame dei nucleoni di valenza e le proprietà dell'alone di molti di questi nuclei possano aumentare o diminuire la probabilità di fusione. La domanda nel comprendere a fondo quale processo (scattering anelastico, reazioni di trasferimento o processo di breakup) sia il principale responsabile dell’aumento della sezione d'urto di reazione alle energie di barriera di Coulomb è stato il filo conduttore di questo lavoro. Nel secondo capitolo sono stati descritti i processi di produzione dei fasci di (_^7)Be e (_^8)B mediante le facility EXOTIC e CRIB. Nel terzo capitolo sono stati decritti i due diversi set up sperimentali usati nelle due reazioni nucleari, in particolare è stata descritta la configurazione dei moduli EXPADES. Nel quarto capitolo sono state trattate tutte le operazioni di Analisi Dati svolte per arrivare alla sezione d’urto differenziale finale per gli isotopi (_^3)He+ (_^4)He prodotti nella reazione (_^7)Be+ (_^208)Pb a 40.5 MeV di energia e il (_^7)Be prodotti nella reazione (_^8)B+ (_^208)Pb ad un'energia di 50 MeV. Nell’ultimo capitolo sono stati discussi i risultati e sono state formulate delle conclusioni

Experimental investigation of the 7Li cosmological problem by means of the Trojan Horse Method applied to the 7Be(n,a)He reaction

With the term “7Li Cosmological Problem” we refer to the discrepancy between the primordial abundance of 7Li, observed in metal-poor halo stars, and the abundance predicted by the Big Bang Nucleo-synthesis (BBN). 7Li is mainly produced from 7Be that undergoes the electron capture process. We thus expect the primordial 7Li abundance to be essentially determined by the 7Be production and destruction rates. In order to find a solution of this problem, in the present work, the case of the 7Be destruction induced by the (n,a) reaction is investigated at the energies typical of the primordial nucleosynthesis by means of the Trojan Horse Method applied to the 2H(7Be,aa)p QF reaction. In the last 5 years many works have been published regarding the 7Be destruction channels involving neutrons, i.e., the 7Be(n,p)7Li, and 7Be(n,α)4He reactions. In particular, the (n,a) reaction channel has been the subject of recent studies. In this work I am going to present a new cross section measurement for the 7Be(n,a)4He performed via the Trojan Horse Method (THM) applied to the Quasi-Free (QF) 2H +7Be reaction. The 7Be(n,a)He cross-section has been measured in a single experiment from 2 MeV down to cosmological energies. The present study represents an important step in the application of THM to reactions between RIBs and neutrons and opens up new windows on many possible future applications. Our result is in remarkable agreement with direct measurements currently available in literature and with the more recent 7Be(n,a)4He measurements discussed in Hayakawa et al. (2020). The values of the 7Be(n,a)4He cross section are in good agreement with those obtained by Lamia et al. (2019) analyzing a complementary set of data collected during the same experiment. Combining both data sets will help in the next future to reduce the uncertainties in the evaluation of the reaction rates in the energy range of interest for the BBN, thus allowing for an overall better characterization of this reaction

Ultrastructural Characterization of Human Bronchial Epithelial Cells during SARS-CoV-2 Infection: Morphological Comparison of Wild-Type and CFTR-Modified Cells

SARS-CoV-2 replicates in host cell cytoplasm. People with cystic fibrosis, considered at risk of developing severe symptoms of COVID-19, instead, tend to show mild symptoms. We, thus, analyzed at the ultrastructural level the morphological effects of SARS-CoV-2 infection on wild-type (WT) and F508del (ΔF) CFTR-expressing CFBE41o- cells at early and late time points post infection. We also investigated ACE2 expression through immune-electron microscopy. At early times of infection, WT cells exhibited double-membrane vesicles, representing typical replicative structures, with granular and vesicular content, while at late time points, they contained vesicles with viral particles. ∆F cells exhibited double-membrane vesicles with an irregular shape and degenerative changes and at late time of infection, showed vesicles containing viruses lacking a regular structure and a well-organized distribution. ACE2 was expressed at the plasma membrane and present in the cytoplasm only at early times in WT, while it persisted even at late times of infection in ΔF cells. The autophagosome content also differed between the cells: in WT cells, it comprised vesicles associated with virus-containing structures, while in ΔF cells, it comprised ingested material for lysosomal digestion. Our data suggest that CFTR-modified cells infected with SARS-CoV-2 have impaired organization of normo-conformed replicative structures

Loss of CFTR Reverses Senescence Hallmarks in SARS-CoV-2 Infected Bronchial Epithelial Cells

SARS-CoV-2 infection has been recently shown to induce cellular senescence in vivo. A senescence-like phenotype has been reported in cystic fibrosis (CF) cellular models. Since the previously published data highlighted a low impact of SARS-CoV-2 on CFTR-defective cells, here we aimed to investigate the senescence hallmarks in SARS-CoV-2 infection in the context of a loss of CFTR expression/function. We infected WT and CFTR KO 16HBE14o-cells with SARS-CoV-2 and analyzed both the p21 and Ki67 expression using immunohistochemistry and viral and p21 gene expression using real-time PCR. Prior to SARS-CoV-2 infection, CFTR KO cells displayed a higher p21 and lower Ki67 expression than WT cells. We detected lipid accumulation in CFTR KO cells, identified as lipolysosomes and residual bodies at the subcellular/ultrastructure level. After SARS-CoV-2 infection, the situation reversed, with low p21 and high Ki67 expression, as well as reduced viral gene expression in CFTR KO cells. Thus, the activation of cellular senescence pathways in CFTR-defective cells was reversed by SARS-CoV-2 infection while they were activated in CFTR WT cells. These data uncover a different response of CF and non-CF bronchial epithelial cell models to SARS-CoV-2 infection and contribute to uncovering the molecular mechanisms behind the reduced clinical impact of COVID-19 in CF patients

R3BRootGroup/R3BRoot: R3BRoot Release November 2023

Framework for Simulations and Data Analysis of R3B Experimen

A new Time-of-flight detector for the R 3 B setup

© 2022, The Author(s).We present the design, prototype developments and test results of the new time-of-flight detector (ToFD) which is part of the R3B experimental setup at GSI and FAIR, Darmstadt, Germany. The ToFD detector is able to detect heavy-ion residues of all charges at relativistic energies with a relative energy precision σΔE/ ΔE of up to 1% and a time precision of up to 14 ps (sigma). Together with an elaborate particle-tracking system, the full identification of relativistic ions from hydrogen up to uranium in mass and nuclear charge is possible.11Nsciescopu