Publications by authors named "Cristiano Salata"

45 Publications

Organoid modeling of Zika and herpes simplex virus 1 infections reveals virus-specific responses leading to microcephaly.

Cell Stem Cell 2021 Apr 7. Epub 2021 Apr 7.

Institute of Molecular Biotechnology (IMBA), Austrian Academy of Sciences, Vienna BioCenter (VBC), Vienna 1030, Austria; Medical University of Vienna, Vienna 1030, Austria. Electronic address:

Viral infection in early pregnancy is a major cause of microcephaly. However, how distinct viruses impair human brain development remains poorly understood. Here we use human brain organoids to study the mechanisms underlying microcephaly caused by Zika virus (ZIKV) and herpes simplex virus (HSV-1). We find that both viruses efficiently replicate in brain organoids and attenuate their growth by causing cell death. However, transcriptional profiling reveals that ZIKV and HSV-1 elicit distinct cellular responses and that HSV-1 uniquely impairs neuroepithelial identity. Furthermore, we demonstrate that, although both viruses fail to potently induce the type I interferon system, the organoid defects caused by their infection can be rescued by distinct type I interferons. These phenotypes are not seen in 2D cultures, highlighting the superiority of brain organoids in modeling viral infections. These results uncover virus-specific mechanisms and complex cellular immune defenses associated with virus-induced microcephaly.
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http://dx.doi.org/10.1016/j.stem.2021.03.004DOI Listing
April 2021

Hypothiocyanite and Hypothiocyanite/Lactoferrin Mixture Exhibit Virucidal Activity In Vitro against SARS-CoV-2.

Pathogens 2021 Feb 19;10(2). Epub 2021 Feb 19.

Department of Molecular Medicine, University of Padova, 35121 Padova, Italy.

SARS-CoV-2 replicates efficiently in the upper airways during the prodromal stage, resulting in environmental viral shedding from patients with active COVID-19 as well as from asymptomatic individuals. There is a need to find pharmacological interventions to mitigate the spread of COVID-19. Hypothiocyanite and lactoferrin are molecules of the innate immune system with a large spectrum cidal activity. The Food and Drug Administration and the European Medicines Agency designated the hypothiocyanite and lactoferrin combination as an orphan drug. We report an in vitro study showing that micromolar concentrations of hypothiocyanite exhibit dose- and time-dependent virucidal activity against SARS-CoV-2 and that the latter is slightly enhanced by the simultaneous presence of lactoferrin.
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http://dx.doi.org/10.3390/pathogens10020233DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7922920PMC
February 2021

Novel Broad-Spectrum Antiviral Inhibitors Targeting Host Factors Essential for Replication of Pathogenic RNA Viruses.

Viruses 2020 12 10;12(12). Epub 2020 Dec 10.

Science for Life Laboratory, Department of Oncology-Pathology, Karolinska Institutet, 171 65 Stockholm, Sweden.

Recent RNA virus outbreaks such as Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and Ebola virus (EBOV) have caused worldwide health emergencies highlighting the urgent need for new antiviral strategies. Targeting host cell pathways supporting viral replication is an attractive approach for development of antiviral compounds, especially with new, unexplored viruses where knowledge of virus biology is limited. Here, we present a strategy to identify host-targeted small molecule inhibitors using an image-based phenotypic antiviral screening assay followed by extensive target identification efforts revealing altered cellular pathways upon antiviral compound treatment. The newly discovered antiviral compounds showed broad-range antiviral activity against pathogenic RNA viruses such as SARS-CoV-2, EBOV and Crimean-Congo hemorrhagic fever virus (CCHFV). Target identification of the antiviral compounds by thermal protein profiling revealed major effects on proteostasis pathways and disturbance in interactions between cellular HSP70 complex and viral proteins, illustrating the supportive role of HSP70 on many RNA viruses across virus families. Collectively, this strategy identifies new small molecule inhibitors with broad antiviral activity against pathogenic RNA viruses, but also uncovers novel virus biology urgently needed for design of new antiviral therapies.
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http://dx.doi.org/10.3390/v12121423DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7762994PMC
December 2020

Hazara virus and Crimean-Congo Hemorrhagic Fever Virus show a different pattern of entry in fully-polarized Caco-2 cell line.

PLoS Negl Trop Dis 2020 11 24;14(11):e0008863. Epub 2020 Nov 24.

Department of Laboratory medicine, Karolinska Institutet, Stockholm, Sweden.

Crimean-Congo Hemorrhagic Fever Virus (CCHFV) and Hazara virus (HAZV) belong to the same viral serotype and family. HAZV has lately been used as a model system and surrogate to CCHFV. However, virus-host cell interaction and level of pathogenicity for these viruses are not well investigated nor compared. In this study, we compared HAZV and CCHFV infection of human polarized epithelial cells to shed light on similarities and differences in virus-host cell interaction between these two viruses. We investigated the pattern of infection of CCHFV and HAZV in fully polarized human cells, the Caco-2 cell line. Polarization of Caco-2 cells lead to difference in expression level and pattern of proteins between the apical and the basolateral membranes. We found that CCHFV virus, in contrast to HAZV, is more likely infecting polarized cells basolaterally. In addition, we found that cytokines/pro-inflammatory factors or other viral factors secreted from CCHFV infected moDC cells enhance the entry of CCHFV contrary to HAZV. We have shown that CCHFV and HAZV early in infection use different strategies for entry. The data presented in this study also highlight the important role of cytokines in CCHFV-host cell interaction.
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http://dx.doi.org/10.1371/journal.pntd.0008863DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7723249PMC
November 2020

Generation of Combinatorial Lentiviral Vectors Expressing Multiple Anti-Hepatitis C Virus shRNAs and Their Validation on a Novel HCV Replicon Double Reporter Cell Line.

Viruses 2020 09 18;12(9). Epub 2020 Sep 18.

Department of Molecular Medicine, University of Padua, 35121 Padua, Italy.

Despite the introduction of directly acting antivirals (DAAs), for the treatment of hepatitis C virus (HCV) infection, their cost, patient compliance, and viral resistance are still important issues to be considered. Here, we describe the generation of a novel JFH1-based HCV subgenomic replicon double reporter cell line suitable for testing different antiviral drugs and therapeutic interventions. This cells line allowed a rapid and accurate quantification of cell growth/viability and HCV RNA replication, thus discriminating specific from unspecific antiviral effects caused by DAAs or cytotoxic compounds, respectively. By correlating cell number and virus replication, we could confirm the inhibitory effect on the latter of cell over confluency and characterize an array of lentiviral vectors expressing single, double, or triple cassettes containing different combinations of short hairpin (sh)RNAs, targeting both highly conserved viral genome sequences and cellular factors crucial for HCV replication. While all vectors were effective in reducing HCV replication, the ones targeting viral sequences displayed a stronger antiviral effect, without significant cytopathic effects. Such combinatorial platforms as well as the developed double reporter cell line might find application both in setting-up anti-HCV gene therapy approaches and in studies aimed at further dissecting the viral biology/pathogenesis of infection.
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http://dx.doi.org/10.3390/v12091044DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7551853PMC
September 2020

Identification and validation of internal reference genes for real-time quantitative polymerase chain reaction-based studies in Hyalomma anatolicum ticks.

Ticks Tick Borne Dis 2020 07 19;11(4):101417. Epub 2020 Mar 19.

Department of Microbiology, The Public Health Agency of Sweden, Nobels Väg 18, SE-171 82 Solna, Sweden; Department for Laboratory Medicine, Karolinska University Hospital and KI, SE-14186 Huddinge, Sweden; National Veterinary Institute, SE-756 51 Uppsala, Sweden. Electronic address:

Crimean-Congo hemorrhagic fever (CCHF) is an emerging tick-borne viral disease caused by the orthonairovirus CCHF virus (CCHFV). Ticks of the genus Hyalomma are the viral reservoir and they represent the main vector transmitting the virus to their hosts during blood feeding. However, how CCHFV replicates in its natural arthropod host cells and the nature of virus/host interactions are still largely unknown. With the aim of developing tools for use in this field, we identified and validated expression of four candidate endogenous control tick genes in a Hyalomma anatolicum-derived cell line. These genes will be useful for normalization of viral/cellular transcripts in infection/expression studies or as internal controls in molecular epidemiology surveys of pathogens transmitted by Hyalomma ticks.
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http://dx.doi.org/10.1016/j.ttbdis.2020.101417DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7284302PMC
July 2020

Coronaviruses: a paradigm of new emerging zoonotic diseases.

Pathog Dis 2019 12;77(9)

Department of Molecular Medicine, University of Padova, Via Gabelli 63, 35121, Padova, Italy.

A novel type of coronavirus (2019-nCoV) infecting humans appeared in Wuhan, China, at the end of December 2019. Since the identification of the outbreak the infection quickly spread involving in one month more than 31,000 confirmed cases with 638 death. Molecular analysis suggest that 2019-nCoV could be originated from bats after passaging in intermediate hosts, highlighting the high zoonotic potential of coronaviruses.
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http://dx.doi.org/10.1093/femspd/ftaa006DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7108526PMC
December 2019

Antiviral treatment and virological monitoring of oseltamivir-resistant influenza virus A(H1N1)pdm09 in a patient with chronic B lymphocytic leukemia.

J Infect Chemother 2019 Jul 20;25(7):543-546. Epub 2019 Apr 20.

Department of Molecular Medicine, University of Padova, Padova, Italy; Microbiology and Virology Unit, Azienda Ospedaliera di Padova, Padova, Italy. Electronic address:

We report the virological monitoring and the antiviral therapy adopted for the treatment of a patient affected by chronic B lymphocytic leukemia, who experienced a severe pneumonia with long-term shedding of influenza virus A(H1N1)pdm09, characterized by an early development of oseltamivir resistance.
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http://dx.doi.org/10.1016/j.jiac.2018.11.008DOI Listing
July 2019

Ebola Virus Entry: From Molecular Characterization to Drug Discovery.

Viruses 2019 03 19;11(3). Epub 2019 Mar 19.

Department of Molecular Medicine, University of Padova, IT-35121 Padova, Italy.

Ebola Virus Disease (EVD) is one of the most lethal transmissible infections, characterized by a high fatality rate, and caused by a member of the family. The recent large outbreak of EVD in Western Africa (2013⁻2016) highlighted the worldwide threat represented by the disease and its impact on global public health and the economy. The development of highly needed anti-Ebola virus antivirals has been so far hampered by the shortage of tools to study their life cycle , allowing to screen for potential active compounds outside a biosafety level-4 (BSL-4) containment. Importantly, the development of surrogate models to study Ebola virus entry in a BSL-2 setting, such as viral pseudotypes and Ebola virus-like particles, tremendously boosted both our knowledge of the viral life cycle and the identification of promising antiviral compounds interfering with viral entry. In this context, the combination of such surrogate systems with large-scale small molecule compounds and haploid genetic screenings, as well as rational drug design and drug repurposing approaches will prove priceless in our quest for the development of a treatment for EVD.
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http://dx.doi.org/10.3390/v11030274DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6466262PMC
March 2019

Sixth European Seminar in Virology on Virus⁻Host Interaction at Single Cell and Organism Level.

Viruses 2018 Jul 28;10(8). Epub 2018 Jul 28.

Department of Molecular Medicine, University of Padova, 35121 Padova PD, Italy.

The 6th European Seminar in Virology (EuSeV) was held in Bertinoro, Italy, 22⁻24 June 2018, and brought together international scientists and young researchers working in the field of Virology. Sessions of the meeting included: virus⁻host-interactions at organism and cell level; virus evolution and dynamics; regulation; immunity/immune response; and disease and therapy. This report summarizes lectures by the invited speakers and highlights advances in the field.
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http://dx.doi.org/10.3390/v10080400DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6116093PMC
July 2018

A first molecular characterization of Listeria monocytogenes isolates circulating in humans from 2009 to 2014 in the Italian Veneto region.

New Microbiol 2018 07 20;41(3):232-234. Epub 2018 Jul 20.

Department of Molecular Medicine, University of Padova, Via Gabelli 63, 35121 - Padova, Italy.

Listeriosis is a disease usually associated with the consumption of low-processed ready-to-eat food products contaminated by Listeria monocytogenes. In Italy, listeriosis has an incidence of 0.19-0.27 cases per 100000 persons. Since detailed information concerning the molecular characterization of listeriosis in the Italian Veneto region is currently lacking, we analyzed 36 L. monocytogenes clinical isolates collected between 2009 and 2014. Results show that the serotype 1/2a was the most represented among the tested samples. No antimicrobial resistance was detected in selected isolates representing the main pulsotypes.
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July 2018

Amiodarone affects Ebola virus binding and entry into target cells.

New Microbiol 2018 Apr 2;41(2):162-164. Epub 2018 Mar 2.

Ebola Virus Disease is one of the most lethal transmissible infections characterized by a high fatality rate. Several research studies have aimed to identify effective antiviral agents. Amiodarone, a drug used for the treatment of arrhythmias, has been shown to inhibit filovirus infection in vitro by acting at the early step of the viral replication cycle. Here we demonstrate that amiodarone reduces virus binding to target cells and slows down the progression of the viral particles along the endocytic pathway. Overall our data support the notion that amiodarone interferes with Ebola virus infection by affecting cellular pathways/ targets involved in the viral entry process.
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April 2018

Antiviral activity of cationic amphiphilic drugs.

Expert Rev Anti Infect Ther 2017 05 20;15(5):483-492. Epub 2017 Mar 20.

a Department of Molecular Medicine , University of Padova , Padova , Italy.

Introduction: Emerging and reemerging viral infections represent a major concern for human and veterinary public health and there is an urgent need for the development of broad-spectrum antivirals. Areas covered: A recent strategy in antiviral research is based on the identification of molecules targeting host functions required for infection of multiple viruses. A number of FDA-approved drugs used to treat several human diseases are cationic amphiphilic drugs (CADs) that have the ability to accumulate inside cells affecting several structures/functions hijacked by viruses during infection. In this review we summarized the CADs' chemical properties and effects on the cells and reported the main FDA-approved CADs that have been identified so far as potential antivirals in drug repurposing studies. Expert commentary: Although there have been concerns regarding the efficacy and the possible side effects of the off-label use of CADs as antivirals, they seem to represent a promising starting point for the development of broad-spectrum antiviral strategies. Further knowledge about their mechanism of action is required to improve their antiviral activity and to reduce the risk of side effects.
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http://dx.doi.org/10.1080/14787210.2017.1305888DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7103695PMC
May 2017

Rapid Bedside Inactivation of Ebola Virus for Safe Nucleic Acid Tests.

J Clin Microbiol 2016 10 27;54(10):2521-9. Epub 2016 Jul 27.

Virus Research & Development Laboratory Statens Serum Institut, Copenhagen, Denmark Infectious Disease Research Unit, University of Southern Denmark, Odense, Denmark.

Rapid bedside inactivation of Ebola virus would be a solution for the safety of medical and technical staff, risk containment, sample transport, and high-throughput or rapid diagnostic testing during an outbreak. We show that the commercially available Magna Pure lysis/binding buffer used for nucleic acid extraction inactivates Ebola virus. A rapid bedside inactivation method for nucleic acid tests is obtained by simply adding Magna Pure lysis/binding buffer directly into vacuum blood collection EDTA tubes using a thin needle and syringe prior to sampling. The ready-to-use inactivation vacuum tubes are stable for more than 4 months, and Ebola virus RNA is preserved in the Magna Pure lysis/binding buffer for at least 5 weeks independent of the storage temperature. We also show that Ebola virus RNA can be manually extracted from Magna Pure lysis/binding buffer-inactivated samples using the QIAamp viral RNA minikit. We present an easy and convenient method for bedside inactivation using available blood collection vacuum tubes and reagents. We propose to use this simple method for fast, safe, and easy bedside inactivation of Ebola virus for safe transport and routine nucleic acid detection.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5035422PMC
http://dx.doi.org/10.1128/JCM.00346-16DOI Listing
October 2016

Full Genome Sequence-Based Comparative Study of Wild-Type and Vaccine Strains of Infectious Laryngotracheitis Virus from Italy.

PLoS One 2016 18;11(2):e0149529. Epub 2016 Feb 18.

Department of Molecular Medicine, University of Padua (DMM), Padua, Italy.

Infectious laryngotracheitis (ILT) is an acute and highly contagious respiratory disease of chickens caused by an alphaherpesvirus, infectious laryngotracheitis virus (ILTV). Recently, full genome sequences of wild-type and vaccine strains have been determined worldwide, but none was from Europe. The aim of this study was to determine and analyse the complete genome sequences of five ILTV strains. Sequences were also compared to reveal the similarity of strains across time and to discriminate between wild-type and vaccine strains. Genomes of three ILTV field isolates from outbreaks occurred in Italy in 1980, 2007 and 2011, and two commercial chicken embryo origin (CEO) vaccines were sequenced using the 454 Life Sciences technology. The comparison with the Serva genome showed that 35 open reading frames (ORFs) differed across the five genomes. Overall, 54 single nucleotide polymorphisms (SNPs) and 27 amino acid differences in 19 ORFs and two insertions in the UL52 and ORFC genes were identified. Similarity among the field strains and between the field and the vaccine strains ranged from 99.96% to 99.99%. Phylogenetic analysis revealed a close relationship among them, as well. This study generated data on genomic variation among Italian ILTV strains revealing that, even though the genetic variability of the genome is well conserved across time and between wild-type and vaccine strains, some mutations may help in differentiating among them and may be involved in ILTV virulence/attenuation. The results of this study can contribute to the understanding of the molecular bases of ILTV pathogenicity and provide genetic markers to differentiate between wild-type and vaccine strains.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0149529PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4758665PMC
July 2016

The Deoxynucleoside Triphosphate Triphosphohydrolase Activity of SAMHD1 Protein Contributes to the Mitochondrial DNA Depletion Associated with Genetic Deficiency of Deoxyguanosine Kinase.

J Biol Chem 2015 Oct 4;290(43):25986-96. Epub 2015 Sep 4.

From the Department of Biology, University of Padova, 35131 Padova, Italy and

The dNTP triphosphohydrolase SAMHD1 is a nuclear antiviral host restriction factor limiting HIV-1 infection in macrophages and a major regulator of dNTP concentrations in human cells. In normal human fibroblasts its expression increases during quiescence, contributing to the small dNTP pool sizes of these cells. Down-regulation of SAMHD1 by siRNA expands all four dNTP pools, with dGTP undergoing the largest relative increase. The deoxyguanosine released by SAMHD1 from dGTP can be phosphorylated inside mitochondria by deoxyguanosine kinase (dGK) or degraded in the cytosol by purine nucleoside phosphorylase. Genetic mutations of dGK cause mitochondrial (mt) DNA depletion in noncycling cells and hepato-cerebral mtDNA depletion syndrome in humans. We studied if SAMHD1 and dGK interact in the regulation of the dGTP pool during quiescence employing dGK-mutated skin fibroblasts derived from three unrelated patients. In the presence of SAMHD1 quiescent mutant fibroblasts manifested mt dNTP pool imbalance and mtDNA depletion. When SAMHD1 was silenced by siRNA transfection the composition of the mt dNTP pool approached that of the controls, and mtDNA copy number increased, compensating the depletion to various degrees in the different mutant fibroblasts. Chemical inhibition of purine nucleoside phosphorylase did not improve deoxyguanosine recycling by dGK in WT cells. We conclude that the activity of SAMHD1 contributes to the pathological phenotype of dGK deficiency. Our results prove the importance of SAMHD1 in the regulation of all dNTP pools and suggest that dGK inside mitochondria has the function of recycling the deoxyguanosine derived from endogenous dGTP degraded by SAMHD1 in the nucleus.
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http://dx.doi.org/10.1074/jbc.M115.675082DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4646252PMC
October 2015

Small RNAs targeting the 5' end of the viral polymerase gene segments specifically interfere with influenza type A virus replication.

J Biotechnol 2015 Sep 16;210:85-90. Epub 2015 Jun 16.

Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy. Electronic address:

Human and avian influenza A viruses, associated with seasonal epidemics and occasionally with pandemics, have a high impact on public health. The development of new antivirals to counteract the emergence of drug resistant influenza virus variants is a main concern. The aim of this study was to develop systems for the efficient and stable expression of small therapeutic RNAs into influenza virus infected cells in order to get further insights on the efficacy of nucleic acid-based antiviral strategies. To this end, lentiviral vectors expressing either microRNAs or antisense-RNAs targeting the 5' end of the PA, PB1 and PB2 influenza virus genomic sequences were generated. Derivative recombinant lentiviral particles were employed to transduce the influenza virus highly susceptible human alveolar basal epithelial A549 cells. The expression of both RNA molecules led to a reduction up to 3 logs of the viral titer when transduced A549 cells were challenged with different human and avian subtypes of influenza type A virus. Importantly, no inhibition of influenza type B virus was observed. Overall our data support the development of nucleic acid-based antiviral strategies to control human and avian influenza A virus infection.
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http://dx.doi.org/10.1016/j.jbiotec.2015.06.391DOI Listing
September 2015

Amiodarone and metabolite MDEA inhibit Ebola virus infection by interfering with the viral entry process.

Pathog Dis 2015 Jul 30;73(5). Epub 2015 Apr 30.

Department of Microbiology, The Public Health Agency of Sweden, Solna 171 82, Sweden Department for Laboratory Medicine, Karolinska Institute, Huddinge/Stockholm 141 83, Sweden National Veterinary Institute, Uppsala 751 89, Sweden.

Ebola virus disease (EVD) is one of the most lethal transmissible infections characterized by a high fatality rate, and a treatment has not been developed yet. Recently, it has been shown that cationic amphiphiles, among them the antiarrhythmic drug amiodarone, inhibit filovirus infection. In the present work, we investigated how amiodarone interferes with Ebola virus infection. Wild-type Sudan ebolavirus and recombinant vesicular stomatitis virus, pseudotyped with the Zaire ebolavirus glycoprotein, were used to gain further insight into the ability of amiodarone to affect Ebola virus infection. We show that amiodarone decreases Ebola virus infection at concentrations close to those found in the sera of patients treated for arrhythmias. The drug acts by interfering with the fusion of the viral envelope with the endosomal membrane. We also show that MDEA, the main amiodarone metabolite, contributes to the antiviral activity. Finally, studies with amiodarone analogues indicate that the antiviral activity is correlated with drug ability to accumulate into and interfere with the endocytic pathway. Considering that it is well tolerated, especially in the acute setting, amiodarone appears to deserve consideration for clinical use in EVD.
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http://dx.doi.org/10.1093/femspd/ftv032DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7108539PMC
July 2015

Multilocus sequence typing of Campylobacter jejuni and Campylobacter coli from humans and chickens in North-Eastern Italy.

New Microbiol 2014 Oct 1;37(4):557-62. Epub 2014 Oct 1.

Department of Comparative Biomedicine and Food Science, University of Padua, Legnaro (PD), Italy.

This paper reports the multilocus sequence typing (MLST) of 57 C. jejuni and C. coli isolates from humans and chickens in Italy and the identification of 17 new sequence types (STs). A high genetic diversity was detected among C. jejuni/C. coli and human/chicken isolates, with a predominance of clonal complexes CC21 and CC828. Although human STs were not the same as those found in chickens, 3 CCs overlapped between human and chicken isolates. Genotyping of Campylobacter strains by MLST should be encouraged in order to implement surveillance and con- trol of infection in humans and in animal reservoirs in Italy.
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October 2014

Antimicrobial resistance of Campylobacter jejuni and Campylobacter coli from poultry in Italy.

Microb Drug Resist 2014 Apr 9;20(2):181-8. Epub 2013 Dec 9.

1 Department of Comparative Biomedicine and Food Science, University of Padua , Legnaro, Italy .

This study was aimed at assessing the antimicrobial resistance (AMR) of Campylobacter isolates from broilers and turkeys reared in industrial farms in Northern Italy, given the public health concern represented by resistant campylobacters in food-producing animals and the paucity of data about this topic in our country. Thirty-six Campylobacter jejuni and 24 Campylobacter coli isolated from broilers and 68 C. jejuni and 32 C. coli from turkeys were tested by disk diffusion for their susceptibility to apramycin, gentamicin, streptomycin, cephalothin, cefotaxime, ceftiofur, cefuroxime, ampicillin, amoxicillin+clavulanic acid, nalidixic acid, flumequine, enrofloxacin, ciprofloxacin, erythromycin, tilmicosin, tylosin, tiamulin, clindamycin, tetracycline, sulfamethoxazole+trimethoprim, chloramphenicol. Depending on the drug, breakpoints provided by Comité de l'antibiogramme de la Société Française de Microbiologie, Clinical and Laboratory Standards Institute, and the manufacturer were followed. All broiler strains and 92% turkey strains were multidrug resistant. Very high resistance rates were detected for quinolones, tetracycline, and sulfamethoxazole+trimethoprim, ranging from 65% to 100% in broilers and from 74% to 96% in turkeys. Prevalence of resistance was observed also against ampicillin (97% in broilers, 88% in turkeys) and at least three cephalosporins (93-100% in broilers, 100% in turkeys). Conversely, no isolates showed resistance to chloramphenicol and tiamulin. Susceptibility prevailed for amoxicillin+clavulanic acid and aminoglycosides in both poultry species, and for macrolides and clindamycin among turkey strains and among C. jejuni from broilers, whereas most C. coli strains from broilers (87.5%) were resistant. Other differences between C. jejuni and C. coli were observed markedly in broiler isolates, with the overall predominance of resistance in C. coli compared to C. jejuni. This study provides updates and novel data on the AMR of broiler and turkey campylobacters in Italy, revealing the occurrence of high resistance to several antimicrobials, especially key drugs for the treatment of human campylobacteriosis, representing a potential risk for public health.
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http://dx.doi.org/10.1089/mdr.2013.0110DOI Listing
April 2014

New generation sequencing in pathogen discovery and microbial surveillance.

Expert Rev Anti Infect Ther 2013 Sep;11(9):877-9

Department of Molecular Medicine, University of Padova, Via Aristide Gabelli 63, 35121 Padova, Italy.

The annual congress of the European Society of Clinical Microbiology and Infectious Diseases (ESCMID) is recognized as the largest European congress for the presentation and discussion of the key priorities and more recent scientific developments in the fields of clinical microbiology and infection. This year, it attracted almost 10,000 participants from all over the world. Keynote lectures, symposia, meet-the-expert sessions, educational workshops, poster and oral sessions covered the diagnosis, treatment, epidemiology and prevention of infectious diseases, as well as related basic microbiology. Moreover, interactive sessions addressing specific subjects underlined the important educational aspect of the ECCMID's congress. The scientific program, abstracts, oral presentations are available at their website [101] . This meeting report is focused on one of the several challenging and one of the most transversal topics of the meeting: the application of the next-generation sequencing (NGS) to the microbial world.
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http://dx.doi.org/10.1586/14787210.2013.827883DOI Listing
September 2013

Absence of class 1 and class 2 integrons among Campylobacter jejuni and Campylobacter coli isolated from poultry in Italy.

J Antimicrob Chemother 2013 Nov 20;68(11):2683-5. Epub 2013 Jun 20.

Department of Comparative Biomedicine and Food Science, University of Padua, Viale dell'Università, 16-35020, Legnaro (Padova), Italy.

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http://dx.doi.org/10.1093/jac/dkt242DOI Listing
November 2013

Human endogenous retroviruses and cancer prevention: evidence and prospects.

BMC Cancer 2013 Jan 3;13. Epub 2013 Jan 3.

Department of Molecular Medicine, Padua University, Padua, Italy.

Background: Cancer is a significant and growing problem worldwide. While this increase may, in part, be attributed to increasing longevity, improved case notifications and risk-enhancing lifestyle (such as smoking, diet and obesity), hygiene-related factors resulting in immuno-regulatory failure may also play a major role and call for a revision of vaccination strategies to protect against a range of cancers in addition to infections.

Discussion: Human endogenous retroviruses (HERVs) are a significant component of a wider family of retroelements that constitutes part of the human genome. They were originated by the integration of exogenous retroviruses into the human genome millions of years ago. HERVs are estimated to comprise about 8% of human DNA and are ubiquitous in somatic and germinal tissues.Physiologic and pathologic processes are influenced by some biologically active HERV families. HERV antigens are only expressed at low levels by the host, but in circumstances of inappropriate control their genes may initiate or maintain pathological processes. Although the precise mechanism leading to abnormal HERVs gene expression has yet to be clearly elucidated, environmental factors seem to be involved by influencing the human immune system.HERV-K expression has been detected in different types of tumors.Among the various human endogenous retroviral families, the K series was the latest acquired by the human species. Probably because of its relatively recent origin, the HERV-K is the most complete and biologically active family.The abnormal expression of HERV-K seemingly triggers pathological processes leading to melanoma onset, but also contributes to the morphological and functional cellular modifications implicated in melanoma maintenance and progression.The HERV-K-MEL antigen is encoded by a pseudo-gene incorporated in the HERV-K env-gene. HERV-K-MEL is significantly expressed in the majority of dysplastic and normal naevi, as well as other tumors like sarcoma, lymphoma, bladder and breast cancer. An amino acid sequence similar to HERV-K-MEL, recognized to cause a significant protective effect against melanoma, is shared by the antigenic determinants expressed by some vaccines such as BCG, vaccinia virus and the yellow fever virus.HERV-K are also reactivated in the majority of human breast cancers. Monoclonal and single-chain antibodies against the HERV-K Env protein recently proved capable of blocking the proliferation of human breast cancer cells in vitro, inhibiting tumor growth in mice bearing xenograft tumors.

Summary: A recent epidemiological study provided provisional evidence of how melanoma risk could possibly be reduced if the yellow fever virus vaccine (YFV) were received at least 10 years before, possibly preventing tumor initiation rather than culling melanoma cells already compromised. Further research is recommended to confirm the temporal pattern of this protection and eliminate/attenuate the potential role of relevant confounders as socio-economic status and other vaccinations.It appears also appropriate to examine the potential protective effect of YFV against other malignancies expressing high levels of HERV-K antigens, namely breast cancer, sarcoma, lymphoma and bladder cancer.Tumor immune-therapy, as described for the monoclonal antibodies against breast cancer, is indeed considered more complex and less advantageous than immune-prevention. Cellular immunity possibly triggered by vaccines as for YFV might also be involved in anti-cancer response, in addition to humoral immunity.
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http://dx.doi.org/10.1186/1471-2407-13-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3557136PMC
January 2013

Tsg101 interacts with herpes simplex virus 1 VP1/2 and is a substrate of VP1/2 ubiquitin-specific protease domain activity.

J Virol 2013 Jan 17;87(1):692-6. Epub 2012 Oct 17.

Department of Molecular Medicine, University of Padova, Padova, Italy.

Ubiquitination/deubiquitination of key factors represent crucial steps in the biogenesis of multivesicular body (MVB) and sorting of transmembrane proteins. We and others previously demonstrated that MVB is involved in herpes simplex virus 1 (HSV-1) envelopment and budding. Here, we report that the HSV-1 large tegument protein, VP1/2, interacts with and regulates the ubiquitination of Tsg101, a cellular protein essential in MVB formation, thus identifying the first cellular substrate of a herpesviral deubiquitinating enzyme.
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http://dx.doi.org/10.1128/JVI.01969-12DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3536428PMC
January 2013

Feline tetherin is characterized by a short N-terminal region and is counteracted by the feline immunodeficiency virus envelope glycoprotein.

J Virol 2012 Jun 18;86(12):6688-700. Epub 2012 Apr 18.

Department of Molecular Medicine, University of Padua, Padua, Italy.

Tetherin (BST2) is the host cell factor that blocks the particle release of some enveloped viruses. Two putative feline tetherin proteins differing at the level of the N-terminal coding region have recently been described and tested for their antiviral activity. By cloning and comparing the two reported feline tetherins (called here cBST2(504) and cBST2*) and generating specific derivative mutants, this study provides evidence that feline tetherin has a shorter intracytoplasmic domain than those of other known homologues. The minimal tetherin promoter was identified and assayed for its ability to drive tetherin expression in an alpha interferon-inducible manner. We also demonstrated that cBST2(504) is able to dimerize, is localized at the cellular membrane, and impairs human immunodeficiency virus type 1 (HIV-1) particle release, regardless of the presence of the Vpu antagonist accessory protein. While cBST2(504) failed to restrict wild-type feline immunodeficiency virus (FIV) egress, FIV mutants, bearing a frameshift at the level of the envelope-encoding region, were potently blocked. The transient expression of the FIV envelope glycoprotein was able to rescue mutant particle release from feline tetherin-positive cells but did not antagonize human BST2 activity. Moreover, cBST2(504) was capable of specifically immunoprecipitating the FIV envelope glycoprotein. Finally, cBST2(504) also exerted its function on HIV-2 ROD10 and on the simian immunodeficiency virus SIVmac239. Taken together, these results show that feline tetherin does indeed have a short N-terminal region and that the FIV envelope glycoprotein is the predominant factor counteracting tetherin restriction.
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http://dx.doi.org/10.1128/JVI.07037-11DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3393548PMC
June 2012

Report of the 2011 annual meeting of the Italian Society for Virology.

J Cell Physiol 2012 Jul;227(7):2965-8

Division of Microbiology and Virology, Department of Molecular Medicine, University of Padova, Padova, Italy.

The 10th annual meeting of the Italian Society for Virology (SIV) comprised seven plenary sessions focused on: General virology and viral genetics; Virus-Host interaction and pathogenesis; Viral oncology; Emerging viruses and zoonotic, foodborne and environmental pathways of transmission; Viral immunology and vaccines; Medical virology and antiviral therapy; Viral biotechnologies and gene therapy. The meeting had an attendance of 143 virologists, about 60% were senior, and the other were young scientists. The submitted abstracts amounted to 88 and the abstracts selected for oral presentation were 41. Complete abstracts of oral and poster presentations are available at the web site www.siv-virologia.it. A summary of the plenary lectures and oral selected presentations is reported.
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http://dx.doi.org/10.1002/jcp.24055DOI Listing
July 2012

Report of two cases of influenza virus A/H1N1v and B co-infection during the 2010/2011 epidemics in the Italian Veneto Region.

Virol J 2011 Nov 3;8:502. Epub 2011 Nov 3.

Department of Histology, Microbiology and Medical Biotechnologies, Division of Microbiology and Virology, University of Padova, Via A, Gabelli 63, Padova 35121, Italy.

From October 2010 to April 2011, in the Italian Veneto Region, 1403 hospitalized patients were tested for influenza virus infection by specific real time RT-PCR. Overall, 327 samples were positive for either influenza A (75%) or B (25%) viruses. Among these positive patients two resulted co-infected by A/H1N1v and B viruses. Even though co-infection with both influenza A and B viruses appears to be a rare event, it occurs naturally and may play a role in epidemiology and pathogenicity. In the present study the two co-infected patients were a transplant recipient immunocompromised adult and a child displaying a severe respiratory illness. The co-infection was confirmed by inoculation of the nasopharyngeal swabs in MDCK.2 cells, followed by immunofluorescence and real time RT-PCR assays. Moreover, in the case of the adult patient, the immune system response against both viruses was assayed by hemoagglutination inhibition test against reference influenza virus strains. Both patients fully recovered from infection, without significant differences with mono-infected patients.
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http://dx.doi.org/10.1186/1743-422X-8-502DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3233533PMC
November 2011

Amiodarone impairs trafficking through late endosomes inducing a Niemann-Pick C-like phenotype.

Biochem Pharmacol 2011 Nov 23;82(9):1234-49. Epub 2011 Aug 23.

Department of Histology, Microbiology and Medical Biotechnologies, University of Padova, via A. Gabelli 63, 35121, Italy.

Patients treated with amiodarone accumulate lysobisphosphatidic acid (LBPA), also known as bis(monoacylglycero)phosphate, in airway secretions and develop in different tissues vacuoles and inclusion bodies thought to originate from endosomes. To clarify the origin of these changes, we studied in vitro the effects of amiodarone on endosomal activities like transferrin recycling, Shiga toxin processing, ESCRT-dependent lentivirus budding, fluid phase endocytosis, proteolysis and exosome secretion. Furthermore, since the accumulation of LBPA might point to a broader disturbance in lipid homeostasis, we studied the effect of amiodarone on the distribution of LBPA, unesterified cholesterol, sphingomyelin and glycosphyngolipids. Amiodarone analogues were also studied, including the recently developed derivative dronedarone. We found that amiodarone does not affect early endosomal activities, like transferrin recycling, Shiga toxin processing and lentivirus budding. Amiodarone, instead, interferes with late compartments of the endocytic pathway, blocking the progression of fluid phase endocytosis and causing fusion of organelles, collapse of lumenal structures, accumulation of undegraded substrates and amassing of different types of lipids. Not all late endocytic compartments are affected, since exosome secretion is spared. These changes recall the Niemann-Pick type-C phenotype (NPC), but originate by a different mechanism, since, differently from NPC, they are not alleviated by cholesterol removal. Studies with analogues indicate that basic pKa and high water-solubility at acidic pH are crucial requirements for the interference with late endosomes/lysosomes and that, in this respect, dronedarone is at least as potent as amiodarone. These findings may have relevance in fields unrelated to rhythm control.
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http://dx.doi.org/10.1016/j.bcp.2011.07.090DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7092840PMC
November 2011