Search

Background. The viral load of asymptomatic SAR-COV-2 positive (ASAP) persons have been equal to that of symptomatic patients, suggesting a similar risk for endothelial dysfunction and increased coagulation in asymptomatic and symptomatic patients. To date, there are no reports of ST-elevation myocardial infarction (STEMI) outcomes in ASAP patients. We evaluated thrombus burden and thrombus viral load and their impact on microvascular bed perfusion in the infarct area (myocardial lush grade, MBG) in ASAP compared to SARS-COV-2 negative (SANE) STEMI patients. Methods. This was an observational study of 46 ASAP, and 130 SANE patients admitted with confirmed STEMI treated with primary percutaneous coronary intervention and thrombus aspiration. The primary endpoints were thrombus dimension + thrombus viral load effects on MBG after PPCI. The secondary endpoints during hospitalization were major adverse cardiovascular events (MACEs). MACEs are defined as a composite of cardiovascular death, nonfatal acute AMI, and heart failure during hospitalization.Results. Thrombus dimensions were significantly higher in ASAP patients as compared to SANE patients. Interestingly, 39 (84.9%) of ASAP patients also had thrombus specimens positive for SARS-COV-2. In ASAP STEMI patients (n=46), thrombus viral load was a significant determinant of thrombus dimension independently of risk factors (pConclusions. In ASAP patients presenting with STEMI, there is strong evidence towards higher thrombus viral load, dimension, and poorer MBG. These data support the need to reconsider ASAP status as a risk factor that may worsen STEMI outcomes.

Conformational changes of viral glycoproteins govern the fusion of viral and cellular membranes in the entry of enveloped viruses. Peptides mimicking domains of viral glycoproteins are apt to interfere with the fusion event, likely hampering the conformational rearrangements from the pre- to the post-fusion structures. We previously developed a peptide sequence with a high potential to inhibit the entry of herpes simplex type 1, which was able to trap glycoprotein B at an intermediate stage, arresting fusion. We propose that similarly to other viruses, membrane targeting through cholesterol conjugation may potently block fusion. The peptide conjugated to polyethylenglycol and cholesterol interacts with viral and cell membranes thanks to the presence of cholesterol and blocks the conformational rearrangements of the glycoprotein B. Here, we also probed the effect of the linker (polyethylenglycol) length on the activity. By targeting the peptide gBh1m to the membranes where fusion occurs and by engineering sequences with increased binding affinity for gB we have enhanced the antiviral potency of our prototype inhibitors. Our results provide proof of concept for the application of cholesterol tagging to develop inhibitors of HSV-1.

// Luca Rinaldi 1 , Veronica Folliero 2 , Luciana Palomba 2 , Carla Zannella 2 , Rachele Isticato 3 , Raffaele Di Francia 4 , Massimiliano Berretta 5 , Ilario de Sio 6 , Luigi E. Adinolfi 1 , Giancarlo Morelli 7 , Secondo Lastoria 8 , Lucia Altucci 6 , Carlo Pedone 7 , Massimiliano Galdiero 2 and Gianluigi Franci 2 1 Department of Medical, Surgical, Neurological, Metabolic and Aging Science, University of Campania “Luigi Vanvitelli”, Naples, Italy 2 Department of Experimental Medicine, University of Campania “Luigi Vanvitelli”, Naples, Italy 3 Department of Biology, Federico II University, Naples, Italy 4 Department of Hematology, National Cancer Institute, Foundation G. Pascale IRCCS, Naples, Italy 5 Department of Medical Oncology, National Cancer Institute IRCCS, Aviano, Italy 6 Department of Precision Medicine, University of Campania “Luigi Vanvitelli”, Naples, Italy 7 Department of Pharmacology, Federico II University, Naples, Italy 8 Department of Diagnostic Imaging, Radiation and Metabolic Therapy, National Cancer Institute, Foundation G. Pascale IRCCS, Naples, Italy Correspondence to: Luca Rinaldi, email: luca.rinaldi@unicampania.it Keywords: microbubbles; sonoporation; ultrasound; liver cancer; gene therapy Received: June 05, 2018 Accepted: July 13, 2018 Published: August 14, 2018 ABSTRACT Introduction: An innovative method, known as sonoporation, was used to induce the expression of silenced genes, such as (but not restricted to) TRAIL and p53, in liver cancer cells (HepG2). The principal aim of the present study was the re-activation of silenced apoptotic pathways in liver cancer models, by using diagnostic synovial microbubble as plasmid gene delivery tools in combination with epigenetic treatments. Material and methods: HepG2 cells were used as a liver cancer model. Microbubbles (Sonovue®) were chosen as gene deliver system in combination with the sonoporation approach. Plasmid pEGFP-TRAIL and pEGFP-p53 were selected and propagated in Escherichia coli grown in LB broth, in order to obtain the necessary amount. Results: Sonoporation was induced by using transducer (Sonitron 2000) and, among the several conditions tested, 3 MHz, 51% Duty Cycle, and 5 W/cm2, 30 s resulted as the best parameters. Data collected showed a dose dependent effect in terms of output energy. A transfection efficacy of 30 – 50% was achieved and recombinant gene expression induced apoptotic effects. In order to increase efficacy, we used the histone deacetylase inhibitor (HDACi, entinostat) MS-275, able to activate TRAIL and thus inducing a stronger pro-apoptotic effect in combination with TRAIL-gene re-expression. Conclusion: For the first time, it was shown the possibility to induce the exogenous expression of the pro-apoptotic gene TRAIL and p53 in a liver cancer HepG2 cells via a sonoporation procedure. The epigenetic treatment using HDACi was able to increase the pro-apoptotic effects of the gene therapy.

The emergence of multidrug resistant bacteria is a global health threat and the discovery of new antimicrobial agents is an absolute priority. In this context endogenous peptides are emerging as novel potential candidates. In this work, we assessed the antimicrobial effects of orexins and ghrelin neuropeptides against gram-negative (Escherichia coli, Salmonella typhimurium, Klebsiella pneumoniae) and gram-positive (Staphylococcus aureus) bacteria. Orexin-B and ghrelin showed a potent bactericidal effect at concentration equal to or greater than 25 μg/ml. No antimicrobial activity has been observed for orexin-A. Furthermore, we investigated the antiviral proprieties of the three peptides against herpes simplex virus 1 (HSV-1). We found that orexin-B, but not orexin-A is effective for HSV-1 infectivity inhibition.

Introduction An innovative method, known as sonoporation, was used to induce the expression of silenced genes, such as (but not restricted to) TRAIL and p53, in liver cancer cells (HepG2). The principal aim of the present study was the re-activation of silenced apoptotic pathways in liver cancer models, by using diagnostic synovial microbubble as plasmid gene delivery tools in combination with epigenetic treatments. Material and methods HepG2 cells were used as a liver cancer model. Microbubbles (Sonovue®) were chosen as gene deliver system in combination with the sonoporation approach. Plasmid pEGFP-TRAIL and pEGFP-p53 were selected and propagated in Escherichia coli grown in LB broth, in order to obtain the necessary amount. Results Sonoporation was induced by using transducer (Sonitron 2000) and, among the several conditions tested, 3 MHz, 51% Duty Cycle, and 5 W/cm2, 30 s resulted as the best parameters. Data collected showed a dose dependent effect in terms of output energy. A transfection efficacy of 30 – 50% was achieved and recombinant gene expression induced apoptotic effects. In order to increase efficacy, we used the histone deacetylase inhibitor (HDACi, entinostat) MS-275, able to activate TRAIL and thus inducing a stronger pro-apoptotic effect in combination with TRAIL-gene re-expression. Conclusion For the first time, it was shown the possibility to induce the exogenous expression of the pro-apoptotic gene TRAIL and p53 in a liver cancer HepG2 cells via a sonoporation procedure. The epigenetic treatment using HDACi was able to increase the pro-apoptotic effects of the gene therapy.

Introduction: An innovative method, known as sonoporation, was used to induce the expression of silenced genes, such as (but not restricted to) TRAIL and p53, in liver cancer cells (HepG2). The principal aim of the present study was the reactivation of silenced apoptotic pathways in liver cancer models, by using diagnostic synovial microbubble as plasmid gene delivery tools in combination with epigenetic treatments. Material and methods: HepG2 cells were used as a liver cancer model. Microbubbles (Sonovue®) were chosen as gene deliver system in combination with the sonoporation approach. Plasmid pEGFP-TRAIL and pEGFP-p53 were selected and propagated in Escherichia coli grown in LB broth, in order to obtain the necessary amount. Results: Sonoporation was induced by using transducer (Sonitron 2000) and, among the several conditions tested, 3 MHz, 51% Duty Cycle, and 5 W/cm2, 30 s resulted as the best parameters. Data collected showed a dose dependent effect in terms of output energy. A transfection efficacy of 30 – 50% was achieved and recombinant gene expression induced apoptotic effects. In order to increase efficacy, we used the histone deacetylase inhibitor (HDACi, entinostat) MS-275, able to activate TRAIL and thus inducing a stronger pro-apoptotic effect in combination with TRAILgene re-expression. Conclusion: For the first time, it was shown the possibility to induce the exogenous expression of the pro-apoptotic gene TRAIL and p53 in a liver cancer HepG2 cells via a sonoporation procedure. The epigenetic treatment using HDACi was able to increase the pro-apoptotic effects of the gene therapy.

In this study we designed and synthesized a new library of antimicrobial peptides correlated to [Pro

In this study we designed and synthesized a new library of antimicrobial peptides correlated to [Pro3,DLeu9]TL 1, a temporin L derivative devoid of cytolytic effects in vitro, and investigated the correlation between the α-helical content of the compounds and their antibacterial, cytotoxic and hemolytic activities. We systematically replaced Gly in position 10 of reference peptide with several amino acids. Structure-activity relationship studies of these analogues were performed by means of antimicrobial and cytotoxicity assays along with CD spectroscopy analyses. NMR analysis was also accomplished for compound 10. As well, the most promising peptides were additionally evaluated for their activity against some clinical strains isolated from human skin and for their mechanism of action by studying the kinetics of membrane perturbation of some representative microbial strains. We identified novel analogues with interesting properties that make them attractive lead compounds for potential topical applications.

HOST DEFENSE TEMPORIN 1 PEPTIDES AND THEIR POTENTIAL AS ANTIVIRAL AGENTS antimicrobial activity on a large number of invading microorganisms. They represent the most ancient and fast-acting elements of the host’s innate defence system against microbial pathogens. Methods: In this scenario, AMPs have emerged as good candidates for the generation of new anti-infective agents. While the antibacterial activity has been deeply investigated, the notions available for antiviral activities are still underexplored. Furthermore, due to their chemical and biological instability only few AMPs are currently in clinical trials as antimicrobial agents. The amphibian temporins represent one of the largest families (more than 100 members) and are among the smallest-sized AMPs (10-16 amino acids) found in nature to date. Temporins are known to be active particularly against Gram-positive bacteria, with minimal inhibitory concentrations ranging from 2.5 μM to 20 μM. In details, we focused on Temporin 1 (TL1) analogues. Results: Those peptides have been modified in order to increase the peptide stability and antimicrobial activity. We then performed a structure-activity relationship study of these peptides with the aim to improve their antimicrobial efficacy without altering their nonhemolytic character. Conclusion: Among the peptides generated, several of them have shown higher in solution stability, low toxicity and a significant decrease of Herpes Simplex virus type 1 infectivity. Our results show novel possible applications of Temporin 1 peptides in the field of antivirals.

Today almost 67% of individuals with age up to 49 years are infected with HSV-1 and 50% of them, with age between 15-49, are infected with genital HSV-1. Naphthoquinones derivatives synthetized by Mannich base derived possess an efficient ability to control in vitro herpesvirus replication. Liposomes are drug delivery systems able to lower the concentrations in order to reduce the toxicity.

Influenza is an ancient and deadly disease which has sickened and killed millions of people in local epidemics and global pandemics. Nowadays, it is common knowledge that influenza is a highly infectious viral illness, but before the discovery of viruses the etiological factor of influenza was not known and, therefore, we had to relay solely on the clinical picture characterized by a sudden onset of high fever, cough, headache, muscle and joint pain, unwell feeling, sore throat, and runny nose. These symptoms were clearly described by Hippocrates roughly 2400 years ago, but historical data on influenza were of difficult interpretation, since these symptoms can be similar to those of other respiratory diseases, therefore not distinctive enough. The word Influenza originated in the 15th century from the Italian language, meaning “influence” since the disease was ascribed to unfavorable astrological influences. A different origin could be the word “influsso” for describing the sweating characteristic of the illness or meaning “influence of the cold.” It was not until 1703 when J. Hugger’s thesis submitted at the University of Edinburgh and named “De Catarrho epidemio, vel Influenza, prout in India occidentali sese ostendit” that the Englishspoken world directly associated “influenza” with the disease and its symptoms. After that the name influenza and its shorthand “flu” came into more general use.1 The influenza virus was first isolated from pigs in 1930 by Shope and Lewis.2 This seminal discovery was followed by the isolation in ferrets of influenza A virus by Smith, Andrewes, and Laidlaw.3 In 1936, Burnet demonstrated that influenza virus could be grown in chicken embryonated eggs,4 opening the path for the study of the characteristics of the virus. It is estimated that influenza virus infects every year 5–10% of the adult population worldwide and 20–30% of the children. Even though most patients recover from flu symptoms within a short periodand without serious sequelae, the estimates indicate from 3–5 million cases of serious illness and over 250,000 deaths per year. Therefore, due to its medical importance, influenza viruses have been the focus of extensive research to decipher the molecular mechanisms that dominate cell invasion and pathogenesis.

Results: Cell cycle analysis was achieved on HepG2 cells transfected with TRAIL-GFP and pEGFP-p53 recombinant protein. Results were analysed with Cell-Quest and ModIFit software. Data shown the re-expression of selected recombinant proteins in over than 30% cells post 24 h from transfection. The transfected cells were treated post 24 h with MS-275 for other 8 h and the cells were collected. The total protein extract was analysed by Western blot and the apoptosis pathways were evaluated via caspase activation proteins. In details we detect the relative bands for caspase 8 and caspase 9 full lengths and activated form in transfected cells and post MS-275 treatment. Conclusion: Results showed the possibility to restore the expression of pro-apoptotic gene TRAIL and p53 in a liver cancer model HepG2. Moreover, the treatment with epigenetic modulators MS-275 enhanced the pro-apoptotic effect mediated by the re-expression of those silenced genes.