27 research outputs found
Characterization of HCV NS5A cellular interactors: role of MOB1B and LRPPRC in viral replication and assembly
No full textHepatitis C virus (HCV) is a hepatotropic positive-strand RNA virus which belongs to the Flaviviridae family. With almost 170 million people infected worldwide, HCV is a leading cause of chronic hepatitis, liver fibrosis, cirrhosis and hepatocellular carcinoma (HCC) (Poynard, T., et al., 2003). Major improvements in patient treatment were achieved by the approval of the first direct acting antivirals (DAAs) in 2011 (Welsch, C., et al., 2012). Nonetheless, HCV variants resistant to DAAs may arise during treatment as well as a heavy burden of side effects, thus affecting therapy outcome. Cellular models of HCV replication and infection have recently allowed the characterization of molecular details of HCV life cycle. HCV RNA replication occurs in association with ER-like cellular membranes and requires several viral non-structural (NS) proteins including; NS3, NS4A, NS4B, NS5A, and NS5B, as well as host cell factors (Moriishi, K., and Matsuura, Y., 2003; Gosert R., et al., 2003). The site of virus assembly is yet unknown, but recent data haves proposed the recruitment of HCV RNA from the replicase complex to lipid droplets mediated by the HCV core and the non-structural proteins as an early event in virion assembly (Miyanari, Y., et al., 2007). Since HCV is a relative small virus, in order to accomplish genome replication and formation of new viral particle, it needs to interact with and subvert cellular machineries for its own purpose. A large number proteinprotein interaction has been described between HCV and host cells. However to date, most of them are only descriptive and their functions in HCV life cycle remain to be characterized (Tellinghuisen and Rice, 2002). NS5A is an important component of the viral replication complex and also participates to HCV particle assembly (Kim, S., et al., 2010). While no known enzymatic function has been ascribed to NS5A, it is known to interact with host cell proteins to affect a variety of processes, including innate immunity, host cell growth and proliferation (He, Y., et al., 2006). The molecular mechanism by which NS5A contributes to HCV life cycles remains largely uncharacterized. To get insights on NS5A function, we performed a screening to search for cellular proteins interacting with NS5A in HCV replicon cells, by means of a tandem affinity purification approach coupled to mass spectrometry (TAPMS/MS). NS5A protein complexes were isolated from HCV replicon cells, resolved by monodimensional gel (SDS-PAGE) and revealed by SYPRORuby staining. Protein bands were submitted to trypsin digestion and identified by MALDI-TOF/TOF mass spectrometry. 24 proteins were identified as NS5A binding proteins, including 14 factors not previously reported. Among them, we functionally characterized the interaction of NS5A with the kinase protein Mob1b, an important regulator of the Hippo pathway, and the mitochondrial protein LRPPRC. Here we show that individually knockdown of Mob1b and LRPPRC significantly reduces new HCV particle production and, to lesser extent, intracellular HCV RNA replication. We also found that Mob1b relocalizes to NS5A-positive structures and Mob1b downregulation results in a decrease in NS5A phosphorylation. Finally, we show that the Hippo signaling pathway is perturbed in HCV replicon cells, as indicated by the increased nuclear localization of the Mob1b target transcription factor Yap. Furthermore we observed that NS5A interaction with LRPPRC occurs at mitochondria-ER synapses (MAM) and that LRPPRC downregulation causes a significant increase in different the inflammation-related genes: IFNβ, Mx1, IL 6 and TNFα. Altogether, our results indicate that, through NS5A interactions, HCV hijacks the cellular functions of Mob1b and LRPPRC to ensure its life cycle
The concept of the mask and the doctor-patient relationship: insights from a (re)reading of Luigi Pirandello’s Six Characters in Search of an Author
No full textThe concept of the mask and the doctor-patient relationship: insights from a (re)reading of Luigi Pirandello's Six Characters in Search of an Author. The aim of this article is to explore the concept of the "mask" and its relevance within the doctor-patient relationship, as well as within the broader medical-scientific context. The primary reference for this analysis is Luigi Pirandello's work Six Characters in Search of an Author (1921), which embodies universal themes, supplemented by examples from figurative art. What emerges is the patient's necessity, utilizing artistic expression, to narrate their own experiences through a mask that serves as the vessel for their innermost emotions
Autophagy in HCV infection: keeping fat and inflammation at bay
No full textHepatitis C virus (HCV) infection is one of the main causes of chronic liver disease. Viral persistence and pathogenesis rely mainly on the ability of HCV to deregulate specific host processes, including lipid metabolism and innate immunity. Recently, autophagy has emerged as a cellular pathway, playing a role in several aspects of HCV infection. This review summarizes current knowledge on the molecular mechanisms that link the HCV life cycle with autophagy machinery. In particular, we discuss the role of HCV/autophagy interaction in dysregulating inflammation and lipid homeostasis and its potential for translational applications in the treatment of HCV-infected patients
The New Organization of Ethics Committees in Italy: What is the Future of Clinical Ethics?
No full textBackground: In Italy, clinical ethics is not well institutionalized. On February 7, 2023, the Italian Ministry of Health published four long-awaited decrees regarding the reorganization of ethics committees. Aim: The aim of this article is twofold: firstly, we aim to briefly summarize the development of clinical ethics in Italy from a legislative point of view; secondly, we aim to examine how Italian regions are implementing the part of the new decrees on the organization of ethics committees that concerns clinical ethics. Methods: As for the first aim, we conducted a critical interpretive review (CIR). The search was restricted to the opinions offered by the Italian National Bioethics Committee (CNB) and to the major Italian legislative decrees on the topic. Regarding the second aim, we conducted an online search through Regional Official Bulletins of each Italian region. Results: Our analysis showed that despite the recommendations from the CNB to differentiate Research Ethics Committees (RECs) and Clinical Ethics Committees (CECs), over the years legislative attention has mainly focused on RECs and pharmacological matters. The new decrees allow regions to be flexible in organizing their activities. However, it emerged that only four regions (Veneto, Friuli-Venezia Giulia, Puglia, Emilia-Romagna) have split the roles, while all the other regions have entrusted both roles to a single committee. Conclusion: The risk for Italy is to take a step backward in the development of clinical ethics. Possible solutions could be either making Local Ethics Committees (CELs) mandatory or institutionalizing Ethics Consultation services (ECSs)
The impact of mevastatin on HCV replication and autophagy of non-transformed HCV replicon hepatocytes is influenced by the extracellular lipid uptake
No full textStatins efficiently inhibit cholesterol synthesis by blocking 3-hydroxy-3-methylglutaryl (HMG)-CoA reductase in the mevalonate pathway. However, the effect of statins on intracellular cholesterol is partially counterbalanced by a consequent increased uptake of extracellular lipid sources. Hepatitis C virus (HCV) infection induces intracellular accumulation of cholesterol by promoting both new synthesis and uptake of circulating lipoproteins, which is required for HCV replication and release. Hepatocytes respond to the increase in intracellular cholesterol levels by inducing lipophagy, a selective type of autophagy mediating the degradation of lipid deposits within lysosomes. In a cellular system of HCV replication based on HuH7 hepatoma cells, statin treatment was shown to be sufficient to decrease intracellular cholesterol, which is accompanied by reduced HCV replication and decreased lipophagy, and has no apparent impact on endocytosis-mediated cholesterol uptake. To understand whether these results were influenced by an altered response of cholesterol influx in hepatoma cells, we analyzed the effect of statins in non-transformed murine hepatocytes (MMHD3) harboring subgenomic HCV replicons. Notably, we found that total amount of cholesterol is increased in MMHD3 cells upon mevastatin treatment, which is associated with increased HCV replication and lipophagy. Conversely, mevastatin is able to reduce cholesterol amounts only when cells are grown in the presence of delipidated serum to prevent extracellular lipid uptake. Under this condition, HCV replication is reduced and autophagy flux is severely impaired. Altogether, these results indicate that both de novo synthesis and extracellular uptake have to be targeted in non-transformed hepatocytes in order to decrease intracellular cholesterol levels and consequently limit HCV replication. Copyright © 2019 Vescovo, Refolo, Manuelli, Tisone, Piacentini and Fimia. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms
Should Artificial Intelligence-Based Patient Preference Predictors Be Used for Incapacitated Patients? A Scoping Review of Reasons to Facilitate Medico-Legal Considerations
No full textBackground: Research indicates that surrogate decision-makers often struggle to accurately interpret and reflect the preferences of incapacitated patients they represent. This discrepancy raises important concerns about the reliability of such practice. Artificial intelligence (AI)-based Patient Preference Predictors (PPPs) are emerging tools proposed to guide healthcare decisions for patients who lack decision-making capacity. Objectives: This scoping review aims to provide a thorough analysis of the arguments, both for and against their use, presented in the academic literature. Methods: A search was conducted in PubMed, Web of Science, and Scopus to identify relevant publications. After screening titles and abstracts based on predefined inclusion and exclusion criteria, 16 publications were selected for full-text analysis. Results: The arguments in favor are fewer in number compared to those against. Proponents of AI-PPPs highlight their potential to improve the accuracy of predictions regarding patients' preferences, reduce the emotional burden on surrogates and family members, and optimize healthcare resource allocation. Conversely, critics point to risks including reinforcing existing biases in medical data, undermining patient autonomy, raising critical concerns about privacy, data security, and explainability, and contributing to the depersonalization of decision-making processes. Conclusions: Further empirical studies are needed to assess the acceptability and feasibility of these tools among key stakeholders, such as patients, surrogates, and clinicians. Moreover, robust interdisciplinary research is needed to explore the legal and medico-legal implications associated with their implementation, ensuring that these tools align with ethical principles and support patient-centered and equitable healthcare practices
From the hands of the robot to the courtroom
No full textAims: The aim of this study was to evaluate the implications of robotic surgery on clinical risk and litigation at the Fondazione Policlinico "A. Gemelli" (FPG) to identify potential strategies to reduce future cases of controversies. Materials and methods: We examined claims related to robotic surgeries performed at FPG from 2015 to 2024. The complication rate for each procedure was evaluated and compared to that of laparoscopic procedures. Subsequently, trends in litigation related to robotic-assisted surgery were analysed, taking into account findings from international literature on the topic. Results: Our data demonstrate a 27% increase in claims, mainly involving Gynecology, Urology, and General Surgery. The primary causes of legal disputes were related to surgical errors and the need for additional procedures. The complication rate was higher in laparoscopic procedures compared to those performed with robotic assistance. Conclusions: The number of claims analysed shows a growing trend, although robotic surgery represents a small proportion of the total number of procedures performed. This finding should encourage healthcare facilities to adopt appropriate tools for assessing and monitoring risks associated with robot-assisted procedures. These tools will help strengthen the skills and training of healthcare workers, as well as improve doctor-patient communication, thus promoting a reduction in claims and an overall improvement in the healthcare experience
Ethical issues in the use of genetic predictions of aggressive behavior in the criminal justice system: a systematic review
No full textBackground: The use of genetic predictions of aggressive behavior in the criminal justice system remains a subject of ongoing debate. Since behavioral genetic evidence is often used in criminal defense arguments, it is crucial to critically examine the ethical challenges associated with its application. Objective: This article seeks to identify and analyze these ethical concerns to ensure the responsible and equitable integration of genetic testing, when deemed necessary, into the judiciary system. Methods: A systematic review was conducted using PubMed, Web of Science, and Scopus, supplemented by manual searches of reference lists to identify additional relevant studies. Results: The search yielded 1,023 publications, 12 of which met the inclusion criteria. Seven key ethical concerns were identified: the risks of discrimination, stigmatization, eugenic reasoning, deterministic interpretations, overestimation of dangerousness, privacy violations, and medicalization, along with the risks posed by limited scientific literacy among legal professionals. Conclusion: The ethical challenges associated with genetic predictions of aggressive behavior underscore the need for a critical and multidisciplinary approach to their use in the criminal justice system. Collaboration among bioethicists, legal scholars, scientists, and communication experts is crucial to prevent misuse and reduce potential biases. Such an approach will help ensure that genetic insights are ethically applied, accurately interpreted, and used to promote justice rather than exacerbate systemic inequalities
The New Italian Guidelines for Assisted Reproduction Technologies (ART): Ethical and Medico-Legal Issues
No full textBackground: On 20 March 2024, the Italian Minister of Health, in collaboration with the Italian National Institute of Health (Istituto Superiore di Sanità) and the National Health Council (Consiglio Superiore di Sanità), issued updated guidelines for assisted reproduction technologies (ART). They introduced two key changes: (1) permitting post-mortem embryo transfers, allowing a woman to proceed with the procedure after her male partner’s death, and (2) permitting embryo transfer even if the male partner is alive but the relationship has ended. Objectives: This study explores the ethical and medico-legal challenges posed by the updated Italian ART Guidelines, with a specific focus on the complexities of informed consent and the ethical dilemmas introduced by these provisions. Methods: This study employs a comprehensive review of legislation, case law, and a comparative analysis of national and international norms. Primary and secondary sources include laws, regulations, court decisions, and key ethical and medico-legal perspectives. Results were organized into two thematic areas: the medico-legal dimension, focusing on informed consent, and the ethical dimension, addressing post-mortem and ex-partner embryo transfers. Results: The new guidelines exacerbate the already intricate ART landscape in Italy. Post-mortem and post-relationship embryo transfers raise significant ethical concerns and amplify legal uncertainties. Conclusions: These updates highlight the pressing need for legislative reform to address unresolved ethical and legal issues in ART
‐19
No full textCoronavirus disease 2019 (COVID-19) is an infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its clinical spectrum ranges from mild to moderate or severe illness. A 78-year-old male was presented at emergency department with dyspnoea, dry cough and severe asthenia. The nasopharyngeal swab by real-time polymerase chain reaction confirmed a SARS-CoV-2 infection. The x-ray and the thoracic ultrasound revealed right pleural effusion. A diagnostic-therapeutic thoracentesis drained fluid identified as chylothorax. Subsequently, the patient underwent a chest computed tomography which showed the radiological hallmarks of COVID-19 and in the following weeks he underwent a chest magnetic resonance imaging to obtain a better view of mediastinal and lymphatic structures, which showed a partial thrombosis affecting the origin of superior vena cava and the distal tract of the right subclavian vein. For this reason, anticoagulant therapy was optimized and in the following weeks the patient was discharged for clinical and radiological improvement. This case demonstrates chylothorax as a possible and uncommon complication of COVID-19
