Search

Hepatitis B virus (HBV) may integrate into the genome of an infected cell and contribute to hepatocarcinogenesis. In this study, we applied a new high-throughput HBV integration sequencing approach that allows sensitive identification of HBV integration sites and enumeration of integration clones. We identified 3,339 HBV integration sites in paired tumour and non-tumour tissue samples from 7 patients with hepatocellular carcinoma (HCC). A total of 2,107 clonally expanded integrations were detected: 1,817 in tumour and 290 in non-tumour tissues. Moreover, we detected significant enrichment of clonal integrations in mitochondrial DNA (mtDNA), and HBV integration in mtDNA preferentially involved OXPHOS genes in tumours and the regulatory D-loop region in non-tumour tissues. We also found that HBV RNA sequences may be imported into the mitochondria of hepatoma cells, polynucleotide phosphorylase (PNPASE) is involved in this import, and HBV RNA might be involved in the process of HBV integration into mtDNA. Our data suggest a novel mechanism by which HBV insertion might contribute to HCC development.

Background & Aims: In the next 20 years, non-alcoholic fatty liver disease (NAFLD) is expected to become the leading cause of liver transplantation. Fibrosis is the most important prognostic factor for liver-related outcomes and mortality, but the need for liver biopsy limits diagnosis. We assessed the performance of plasma ficolin-2 (FCN-2) as a biomarker of fibrosis. FCN-2 candidate was selected by an in silico discovery strategy. Methods: We enrolled 235 morbidly obese (MO) subjects with biopsy-proven NAFLD stratified by fibrosis stage (F0, n=44; F1, n=134; F2, n=46; F3/F4, n=11) and 40 cirrhotic patients as positive controls. The cohort was subdivided into discovery (n=76) and validation groups (n=159). The plasma level of FCN-2 and other biomarkers was determined by enzyme-linked immunoabsorbent assay. Results: Plasma level of FCN-2 correlated inversely with the stage of liver fibrosis (ρ = -0.49, pConclusion: FCN-2 plasma level can accurately discriminate liver fibrosis status (minimal vs . moderate/advanced). Its inclusion in the diagnostic workup significantly improves the fibrosis diagnostic algorithms.

Fibrosis is the strongest predictor for disease-specific mortality in non-alcoholic fatty liver diseases (NAFLD), but the need for liver biopsy limits its diagnosis. We assessed the performance of plasma ficolin-2 (FCN-2) as a biomarker of fibrosis identified by an in silico discovery strategy. Two hundred and thirty-five morbidly obese (MO) subjects with biopsy-proven NAFLD stratified by fibrosis stage (F0, n = 44; F1, n = 134; F2, n = 46; F3/F4, n = 11) and 40 cirrhotic patients were enrolled. The cohort was subdivided into discovery (n = 76) and validation groups (n = 159). The plasma level of FCN-2 and other candidate markers was determined. FCN-2 was inversely correlated with the stage of liver fibrosis (ρ = −0.49, p < 0.001) independently of steatosis (p = 0.90), inflammation (p = 0.57), and ballooning (p = 0.59). In the global cohort, FCN-2 level decreased significantly in a stepwise fashion from F0/F1 (median 4753 ng/mL) to F2–F3–F4 (2760 ng/mL) and in cirrhotic subjects (1418 ng/mL). The diagnostic performance of FCN-2 in detecting F ≥ 2 was higher than other indexes (APRI, FIB-4) (AUROC 0.82, 0.68, and 0.6, respectively). The accuracy improved when combined with APRI score and HDL values (FCNscore, AUROC 0.85). Overall, the FCN-2 plasma level can accurately discriminate liver fibrosis status (minimal vs. moderate/advanced) significantly improving the fibrosis diagnostic algorithms.

Introduction: Measles is a contagious disease that re-emerged among young adults as a consequence of suboptimal vaccination coverage. Since in the pre-vaccination era measles affected mainly children, little is known about measles-associated respiratory complications in adults. The aim of this study was to describe clinical and radiological findings in adults affected by measles who developed respiratory complications during a recent measles outbreak. Material and methods: In this retrospective chart review-based study we analyzed data from patients admitted for measles from January to June 2018 to a large tertiary care hospital, in one of the main cities in the south of Italy. This city has been the country’s heart of the epidemic with a high morbidity and mortality rate. Results: Among 177 patients (mean age 26 ± 9 years), only 2 were vaccinated. Thirty patients (16.9%) had signs of pneumonia on chest radiography. Computed tomography scan showed the following abnormalities: centrilobular nodules (63%), ground-glass attenuation (63%), air-space consolidation (36%), pleural effusion (16%) and pneumothorax (10%). Five patients developed severe lung injury and hypoxemia requiring admission to Intensive Care Unit. Two young unvaccinated women with no past medical history died from acute respiratory failure. The death was sudden and unpredictable. Conclusions: Measles-associated pneumonia in unvaccinated young adults can cause severe respiratory impairment and death. Our findings support the need for a mandatory vaccination policy.

Background & aims Obesity is associated with non-alcoholic fatty liver (NAFL), which may progress towards non-alcoholic steatohepatitis (NASH), cirrhosis and hepatocellular carcinoma (HCC). Occult hepatitis B virus infection (OBI) may contribute to hepatic damage in patients with chronic liver disease of different aetiologies (eg HCV, alcohol). However, information on the prevalence and clinical impact of OBI in obese individuals is lacking. The aims of this study were to investigate NASH prevalence and risk factors in obese people who underwent bariatric surgery. Methods Two-hundred and twenty-six subjects (160 females; mean age 42.9 years ±10.8 SD) without evidence of any further cause of liver disease consecutively underwent bariatric surgery in two Italian liver centers. During surgery, all patients underwent liver biopsy for histological evaluation and molecular studies. Liver DNA extracts were tested for PNPLA3, TM6SF2, MBOAT7, IRGM polymorphisms and for OBI. Univariate and multivariate analyses were used to identify predictors of NASH. Results Histology showed NASH in 115 (50.9%) and NAFL in 111 cases (49.1%). Twenty-nine/226 (12.8%) cases had OBI, 24 (82.8%) of whom had NASH and 5 (17.2%) NAFL, whereas among the 197 OBI-negative cases, 91 (46.2%) had NASH and 106 (53.8%) NAFL (P = .0002). Multivariate analysis showed that older age (P = .03, OR 1.034), alanine aminotransferase values (P = .005, OR 1.023), insulin resistance/diabetes (P = .02, OR 2.257), TM6SF2 polymorphism (P = .04, OR 3.168) and OBI (P = .004, OR 5.503) were independent predictors of NASH. Conclusion NASH is highly prevalent in obese individuals undergoing bariatric surgery. OBI is one of the strongest risk factors of NASH in these patients.

Negative thoughts are experienced by as many as 80–99% of the population. These thoughts are associated with a variety of negative consequences, including negative mood, decreased task performance and the development of psychopathology. One technique employed in contextual behavioral therapies to help cope with negative thoughts is cognitive defusion. Cognitive defusion techniques undermine potential negative effects of thinking by teaching clients to get some distance from their thoughts. Virtual reality (VR) is the computer-generated simulation of a three-dimensional environment that users can interact in. VR is of increasing interest to applied psychologists due to its potential for exposure learning. One area where VR may be effective is helping people to cope with negative thoughts. The current study examined the impact of a VR task as a cognitive defusion technique on participants’ relationship with a negative self-referential thought (e.g., “I am a failure”). Thirty participants were randomly assigned to one of two conditions (i.e., defusion VR and control VR). Participants were tested pre- and post-VR task on a state measure of cognitive defusion and ratings of their self-referential negative thought. The results indicated that a defusion VR task facilitates the management of negative thoughts and leads to an increase in state defusion. The findings are discussed in terms of their implications for the use of VR techniques in dealing with negative thoughts.

The objective of our research was to compare the initial efficacy and acceptability, in this special population, of different motivational stimuli to improve motivation to quit smoking. We enrolled 40 young adult smokers, between their twenties and their thirties, not motivated to quit. Our study adopted a visual analog scale to assess the motivation to quit at first visit (baseline) and after three different types of stimuli: a packet of cigarettes containing shocking images; a brief film showing pulmonary effects of smoking; a virtual reality period based on the progression of smoking-related illnesses. We compared the motivation to quit smoking between the series of stimuli and between the stimuli and the baseline assessment. Compared with the first assessment, all stimuli were significant in improving the motivation for smoking cessation. There was a statistically significant modification between the packet of cigarettes and the movie. The difference between the packet of cigarettes containing shocking images and the virtual reality was significant. The difference between the film and the virtual reality was also highly significant. The application of virtual reality would appear to greatly increase the motivation to smoking cessation in young adult smokers not motivated to quit.

Background There is controversial data on possible occult HBV reactivation in HCV patients successfully treated with direct-acting antivirals (DAA). However, diagnosis of occult HBV infection (OBI) was not performed by gold standard procedures in any study. Methods By using several highly sensitive assays, we examined serially collected serum samples from 40 HBV-surface-negative DAA-treated HCV patients with OBI identified by testing liver biopsy specimens through nested-PCR technique. Serum samples were obtained at four time points from each patient (at baseline, at 4 weeks after starting, at the end and 12 weeks after stopping therapy) and tested for HBV DNA by nested-PCR and real-time PCR techniques. Results All tested serum samples were negative by both quantitative HBV surface antigen (HBsAg) and HBV core-related antigen assays. 26/40 patients were anti-HBs-positive and in all of them, the amount of this antibody was stable at the four time points evaluated. Serum HBV DNA was detected in 10 samples at baseline, in 6 samples 4 weeks after starting therapy, in 11 samples at the end of therapy and in 21 samples 12 weeks after stopping treatment ( P=0.001). Aminotransferase values dropped within the normal levels at week 4 of therapy and persisted normal over time in all cases. Conclusions A slight increase in the amount of HBV DNA 3 months after stopping DAA therapy was the only parameter showing a possible reappearance of HBV activity in OBI patients cured for a concomitant HCV infection, but it was insufficient to lead toward a virological reactivation capable of inducing liver injury.

Flavocoxid is a proprietary blend of two flavonoids, baicalin and catechin, and recent evidence has shown that bioflavonoids may exert antiviral activities. The potential antiviral activity of Flavocoxid against hepatitis B virus (HBV) was evaluated. Additionally, it was investigated if Flavocoxid used in combination with Entecavir could potentiate its anti-HBV activity. Hepatoma cells replicating HBV were treated with Flavocoxid, or Entecavir alone or in combination for up to 5 days. Viral replicative intermediates, transcripts, and cccDNA levels were evaluated in HBV-replicating cells by real-time PCR, Southern and Northern blotting. Expression profiling was performed using TaqMan low-density arrays. Flavocoxid treatment induced a reduction of HBV replicative intermediates, the amount of transcripts, and HBsAg levels. Flavocoxid and Entecavir combination therapy further decreased the amount of HBV replicative intermediates, compared to Flavocoxid alone. Importantly, Flavocoxid alone or in combination with Entecavir also induced a reduction of cccDNA. Gene-expression analysis showed that Flavocoxid activates type I IFNs-signaling and dampens the HBV-induced inflammatory response. Flavocoxid inhibits HBV replication by targeting multiple steps of viral life cycle. These results indicate that the antiviral activity of Entecavir is potentiated by Flavocoxid, suggesting that this medical food might be considered as an adjuvant for anti-HBV therapy.

Protein induced by vitamin K absence-II (PIVKA-II) is a potential screening marker for hepatocellular carcinoma (HCC). Limited data are available about its utility in discriminating neoplastic from regenerative nodules at ultrasonography (US) evaluation in cirrhotic patients. Aim of this study was to investigate the diagnostic utility of PIVKA-II in cases showing liver nodules of uncertain diagnosis at US. Ninety cirrhotics with US evidence of liver nodule(s) were enrolled. All patients underwent blood sampling within 1 week of US and were thereafter followed up. HCC was confirmed in 40/90 cases, and in all cases it was in a very early/early stage. All sera were tested for PIVKA-II and alpha-fetoprotein (AFP) at the end of follow-up. PIVKA-II at a cut off of 60 mAU/mL was significantly associated with HCC at both univariate and multivariate analysis (P = .016 and P = .032, respectively). AFP at a cut off of 6.5 ng/mL was not associated with HCC at univariate analysis (P = .246). ROC curves showed that PIVKA-II had 60% sensitivity, 88% specificity, 80% positive predictive value (PPV), and 73% negative predictive value (NPV), whereas AFP had 67% sensitivity, 68% specificity, 63% PPV, and 72% NPV. AUROC curves showed that the combination of both biomarkers increased the diagnostic accuracy for HCC (AUC 0.76; sensitivity 70%, specificity 94%, PPV 91%, and NPV 79%). In conclusion, PIVKA-II is a useful tool for the diagnostic definition of US-detected liver nodules in cirrhotic patients, and it provides high diagnostic accuracy for HCC when combined with AFP.

Autophagy is a cellular degradation pathway that exerts numerous functions in vital biological processes. Among these, it contributes to both innate and adaptive immunity. On the other hand, pathogens have evolved strategies to manipulate autophagy for their own advantage. By monitoring autophagic markers, we showed that HSV-1 transiently induced autophagosome formation during early times of the infection of monocytic THP-1 cells and human monocytes. Autophagy is induced in THP-1 cells by a mechanism independent of viral gene expression or viral DNA accumulation. We found that the MyD88 signaling pathway is required for HSV-1-mediated autophagy and it is linked to the toll-like receptor 2 (TLR2). Interestingly, autophagy inhibition by pharmacological modulators or siRNA knockdown impaired viral replication in both THP-1 cells and human monocytes, suggest that the virus exploits the autophagic machinery to its own benefit in these cells. Taken together, these findings indicate that the early autophagic response induced by HSV-1 exerts a proviral role, improving viral production in a semi-permissive model such as THP-1 cells and human monocytes.

Background: Perioperative atrial tachyarrhythmias (POAT) are the commonest arrhythmia after major lung resections. To date the relationship between POAT and CPET parameters were never tested after major thoracic surgery. Aim: To assess the association between CPET data and postoperative complications with particular focus on POAT. Methods: 48 consecutive patients undergoing major lung resection from Jan to Dec 2013 were assessed by pre-operative spirometry, arterial blood gas analysis, DLCO, CPET on cycle ergometer, ECG and echocardiogram. Cardiologic and respiratory complications were recorded and matched with functional parameters and to identify any potential risk factor. Results: Overall morbidity, POAT, atrial fibrillation (AF) and respiratory failure incidences were 18 (37.5%), 10 (20.8%), 5 (12.8%) and 9 (18%) respectively. The followed associations were found: older age (p=0.048), steeper Ve/VCO2 slope (p=0.002) and lower Maximum Heart Rate (MRH) (p=0.011) with POAT. Regression analysis indicated higher Ve/VCO2 (p=0.011; HR=1.39[CI95%:1.07-1.69],) and lower FC max (p=0.022; HR=1.06[CI95%:1.01-1.11],) as independent risk factors for POAT. AF was associated with age (p=0.037), lower VO2 max ml/Kg/min %pred (p=0.02), Ve/VCO2 slope (p=0.01), MHR (p=0.001); Pulmonary artery pressure (p=0.016); VO2 max ml/Kg/min %pred (p=0.011; HR=1.09[CI95%:1.02-1.16],) and lower MHR (p=0.048;HR=1.07[CI95%:1.00-1.15]) predict the risk of AF. Conclusions: VE/VCO2, MHR, VO2 max ml/Kg/min %pred are objective good predictor of POAT and AF and should be helpful to select patients for prophylactic therapy.

Background: Mount Etna, located in the eastern part of Sicily (Italy), is the highest and most active volcano in Europe. During the sustained eruption that occurred in October-November 2002 huge amounts of volcanic ash fell on a densely populated area south-east of Mount Etna in Catania province. The volcanic ash fall caused extensive damage to infrastructure utilities and distress in the exposed population. This retrospective study evaluates whether or not there was an association between ash fall and acute health effects in exposed local communities. Methods: We collected the number and type of visits to the emergency department (ED) for diseases that could be related to volcanic ash exposure in public hospitals of the Province of Catania between October 20 and November 7, 2002. We compared the magnitude of differences in ED visits between the ash exposure period in 2002 and the same period of the previous year 2001. Results: We observed a significant increase of ED visits for acute respiratory and cardiovascular diseases, and ocular disturbances during the ash exposure time period. Conclusions: There was a positive association between exposure to volcanic ash from the 2002 eruption of Mount Etna and acute health effects in the Catania residents. This study documents the need for public health preparedness and response initiatives to protect nearby populations from exposure to ash fall from future eruptions of Mount Etna.