Post-translational deregulation of YAP1 is genetically controlled in rat liver cancer and determines the fate and stem-like behavior of the human disease

Previous studies showed that YAP1 is over-expressed in hepatocellular carcinoma (HCC). Here we observed higher expression of Yap1/Ctgf axis in dysplastic nodules and HCC chemically-induced in F344 rats, genetically susceptible to hepatocarcinogenesis, than in lesions induced in resistant BN rats. In BN rats, highest increase in Yap1-tyr357, p73 phosphorylation and Caspase 3 cleavage occurred. In human HCCs with poorer prognosis ( 3 years survival; HCCB). In the latter, higher levels of phosphorylated YAP1-ser127, YAP1-tyr357 and p73, YAP1 ubiquitination, and Caspase 3 cleavage occurred. Expression of stemness markers NANOG, OCT-3/4, and CD133 were highest in HCCP and correlated with YAP1 and YAP1-TEAD levels. In HepG2, Huh7, and Hep3B cells, forced YAP1 over-expression led to stem cell markers expression and increased cell viability, whereas inhibition of YAP1 expression by specific siRNA, or transfection of mutant YAP1 which does not bind to TEAD, induced opposite alterations. These changes were associated, in Huh7 cells transfected with YAP1 or YAP1 siRNA, with stimulation or inhibition of cell migration and invasivity, respectively. Furthermore, transcriptome analysis showed that YAP1 transfection in Huh7 cells induces over-expression of genes involved in tumor stemness. In conclusion, Yap1 post-translational modifications favoring its ubiquitination and apoptosis characterize HCC with better prognosis, whereas conditions favoring the formation of YAP1-TEAD complexes are associated with aggressiveness and acquisition of stemness features by HCC cells

Ebola virus disease: In vivo protection provided by the PAMP restricted TLR3 agonist rintatolimod and its mechanism of action

Ebola virus (EBOV) is a highly infectious and lethal pathogen responsible for sporadic self-limiting clusters of Ebola virus disease (EVD) in Central Africa capable of reaching epidemic status. 100% protection from lethal EBOV-Zaire in Balb/c mice was achieved by rintatolimod (Ampligen) at the well tolerated human clinical dose of 6 mg/kg. The data indicate that the mechanism of action is rintatolimod's dual ability to act as both a competitive decoy for the IID domain of VP35 blocking viral dsRNA sequestration and as a pathogen-associated molecular pattern (PAMP) restricted agonist for direct TLR3 activation but lacking RIG-1-like cytosolic helicase agonist properties. These data show promise for rintatolimod as a prophylactic therapy against human Ebola outbreaks

Trapping polarization of light in nonlinear optical fibers: An ideal Raman polarizer

The main subject of this contribution is the all-optical control over the state of polarization (SOP) of light, understood as the control over the SOP of a signal beam by the SOP of a pump beam. We will show how the possibility of such control arises naturally from a vectorial study of pump-probe Raman interactions in optical fibers. Most studies on the Raman effect in optical fibers assume a scalar model, which is only valid for high-PMD fibers (here, PMD stands for the polarization-mode dispersion). Modern technology enables manufacturing of low-PMD fibers, the description of which requires a full vectorial model. Within this model we gain full control over the SOP of the signal beam. In particular we show how the signal SOP is pulled towards and trapped by the pump SOP. The isotropic symmetry of the fiber is broken by the presence of the polarized pump. This trapping effect is used in experiments for the design of new nonlinear optical devices named Raman polarizers. Along with the property of improved signal amplification, these devices transform an arbitrary input SOP of the signal beam into one and the same SOP towards the output end. This output SOP is fully controlled by the SOP of the pump beam. We overview the sate-of-the-art of the subject and introduce the notion of an "ideal Raman polarizer"

Children and Families' mental health during the first COVID-19 lockdown in Italy

Background: This study aimed to screen a wide range of emotional and behavioural variables emerging during the first COVID-19 pandemic-lockdown in a sample of parents and children, residents in the southern part of Italy, and explore which variables could predict children's wellbeing. We hypothesised that difficulties in adapting routines to pandemic restrictions, parents' emotional wellbeing, and attitude towards the pandemic could influence the children's behavioural attitudes. Methods: 221 parents completed the survey and gave information about 246 children. Ad hoc questionnaires were created and then exploratory reduced in factors. Strengths and Difficulties Questionnaire (SDQ) for parents assessed positive and negative behavioural attitudes in children. Depression Anxiety Stress Scale (Italian DASS-21) scored depression, anxiety and stress in parents. Results: Children presented higher emotional distress (Mean difference (Mdiff)=0.6, 95% C.I. 0.2, 0.9, p=0.013) and better prosocial behaviour (Mdiff=0.5, 95% C.I. 0.1, 0.9, p=0.011) than the Italian normative sample. Parents were more depressed than expected in the general population (Mdiff=1.0, 95% C.I. 0.3, 1.6, p=0.005). Having developed a morbid attachment to an adult (B=0.37, 95% CI 0.05, 0.69, p=0.024), a higher parental depression (B=0.1, 95% CI 0.02, 0.18, p=0.014), and children's suffering from nightmares (B=0.35, 95% CI 0.03, 0.67, p=0.032) explained the 31.9% of the total variance in children's emotional distress. Children's anxiety was related to parents' fear of the pandemic effects (r=0.32, p=0.001) and avoiding communicative approach (r=0.24, p=0.011). Conclusion: The first lockdown determined emotional distress and regressive mechanisms in children in the contest of higher parental discomfort, fear of the infection and avoidant communication. Following parents' indications, it could be helpful to provide families with informative and age-appropriate support

The role of different game-genres in predicting internet gaming disorder (IGD)

Introduction: Internet gaming disorder (IGD) is a new diagnosis in DSM 5 worth of research. New potentially addictive features are emerging in pay- and free-to-play videogames, involving different at-risk populations of gamers. However, few studies have examined whether and how different game-genres can contribute to the risk of IGD. Objectives: This study aimed to investigate how game-genres can predict IGD, accounting for alexithymia scores, time-related play- ing habits, and other predictors. Methods: Participants were gamers joining online communities, surveyed about which games they played more than 20 hours in their lifetime, time-variables, other stressors and alexithymia scores. A six-steps linear regression with IGD scores and a post hoc logistic regression (outcome: IGD>=21) were performed. Results: 5,979 subjects (88.7% males, 14-18 years), playing at different games (Figure-1). The game-genre explained the 1% of variation only. WoW and similar MMORPGs confirmed their potentiality in promoting IGD, regardless of alexithymia features (B=0.50, p=0.005). However, time-variables completely absorbed the WoW effect (B=0.01, p=0.951). LoL resulted addictive, even if considering time-variables and alexithymia (B=0.88, p<0.001). Minecraft emerged when time-variables were inserted (B=0.359, p=0.041) and stayed significant if removing alexithymia scores (B=0.48, p=0.010). Playing at Diablo3 and similar RPG did not increase IGD (B=-0.99, p>0.001). None of the different game- genres was able to push the subject over the threshold of IGD, because other characteristics interacted as additive risk-factors. Conclusions: Alexithymia traits and time-related playing habits mostly moderated the effect of different games in increasing IGD risk. A videogame could engage people with specific characteristics that may, in turn, differentially predispose to IGD

Differences between female and male gamers and gender-specific risk-factors for internet gaming disorder (IGD)

Introduction: Videogames have become more popular across females, although their widespread diffusion among males. How- ever, few studies have examined differences between female and male gamers and gender-specific risk factors for Internet Gaming Disorder (IGD). Objectives: The study aimed to describe males and females’ differ- ences in a sample of gamers, and to identify gender-specific risk- factors for IGD, accounting for alexithymia, playing habits, and other perceived stressors. Methods: Participants were gamers joining online communities, tested by IGDS-SF9 and TAS-20 for alexithymia. To explore isk-factors for IGD (outcome: IGD>=21), we set a binary logistic regression stratified by gender. Results: 5,305 males and 674 females differed in most of the descrip- tive characteristics (Figure-1) and game-genres preferences (Figure- 2). Higher DIF scores increased the risk of IGD in both males (OR=1.8 95% C.I. 1.6, 2) and females (OR=1.3 95% C.I. 1.1, 1.7) while higher EOT in males only (OR=1.2 95% C.I. 1.1, 1.3). Having another hobby apart from gaming was protective for males (OR=0.5, 95% C.I. 0.4, 0.6). Having started playing before their ten-years was a risk factor for females (OR=2.3 95% C.I. 1.2, 4.6). Loneliness and boredom feelings predicted IGD in males (OR=1.7 95% C.I. 1.5, 2) and, even more, in females (OR=2.7 95% C.I. 1.8, 4.2). Playing more than six hours/per day increased IGD-risk up to seven times in males (OR=7.3 95% C.I. 5.1, 10.3) and of almost sixteen times in females (OR=15.9 95% C.I. 5.4, 46.7) (Figure-3). Conclusions: Female gamers presented specific characteristics and a greater vulnerability to the increased time spent playing as a risk- factor for IG

Mapping a Sex Hormone–Sensitive Gene Determining Female Resistance to Liver Carcinogenesis in a Congenic F344.BN-Hcs4 Rat

Hepatocellular carcinoma (HCC) is prevalent in human and rodent males. Hepatocarcinogenesis is controlled by various genes in susceptible F344 and resistant Brown Norway (BN) rats. B alleles at Hcs4 locus, on RNO16, control neoplastic nodule volume. We constructed the F344.BN-Hcs4 recombinant congenic strain (RCS) by introgressing a 4.41-cM portion of Hcs4 from BN strain in an isogenic F344 background. Preneoplastic and neoplastic lesions were induced by the ''resistant hepatocyte'' protocol. Eight weeks after initiation, lesion volume and positivity for proliferating cell nuclear antigen (PCNA) were much higher in lesions of F344 than BN rats of both sexes. These variables were lower in females than in males. Lesion volume and PCNA values of male RCS were similar to those of F344 rats, but in females corresponded to those of BN females. Carcinomatous nodules and HCC developed at 32 and 60 weeks, respectively, in male F344 and congenics and, rarely, in F344 females. BN and congenic females developed only eosinophilic/clear cells nodules. Gonadectomy of congenic males, followed by B-estradiol administration, caused a decrease in Ar expression, an increase in Er-a expression, and development of preneoplastic lesions comparable to those from BN females. Administration of testosterone to gonadectomized females led to Ar increase and development of preneoplastic lesions as in F344 males. This indicates a role of homozygous B alleles at Hcs4 in the determination of phenotypic patterns of female RCS and presence at Hcs4 locus of a high penetrance gene(s), activated by estrogens and inhibited/unaffected by testosterone, conferring resistance to females in which the B alleles provide higher resistance. (Cancer Res 2006; 66(21): 10384-90

Retargeted adenoviruses for radiation-guided gene delivery

The combination of radiation with radiosensitizing gene delivery or oncolytic viruses promises to provide an advantage that could improve the therapeutic results for glioblastoma. X-rays can induce significant molecular changes in cancer cells. We isolated the GIRLRG peptide that binds to radiation-inducible 78 kDa glucose-regulated protein (GRP78), which is overexpressed on the plasma membranes of irradiated cancer cells and tumor-associated microvascular endothelial cells. The goal of our study was to improve tumor-specific adenovirus-mediated gene delivery by selectively targeting the adenovirus binding to this radiation-inducible protein. We employed an adenoviral fiber replacement approach to conduct a study of the targeting utility of GRP78-binding peptide. We have developed fiber-modified adenoviruses encoding the GRP78-binding peptide inserted into the fiber-fibritin. We have evaluated the reporter gene expression of fiber-modified adenoviruses in vitro using a panel of glioma cells and a human D54MG tumor xenograft model. The obtained results demonstrated that employment of the GRP78-binding peptide resulted in increased gene expression in irradiated tumors following infection with fiber-modified adenoviruses, compared with untreated tumor cells. These studies demonstrate the feasibility of adenoviral retargeting using the GRP78-binding peptide that selectively recognizes tumor cells responding to radiation treatment

aberrant inos signaling is under genetic control in rodent liver cancer and potentially prognostic for the human disease

Mounting evidence underlines the role of inducible nitric oxidesynthase (iNOS) in hepatocellular carcinoma (HCC) develop-ment, but its functional interactions with pathways involved inHCC progression remain uninvestigated. Here, we analyzed inpreneoplastic and neoplastic livers from Fisher 344 and BrownNorway rats, possessing different genetic predisposition to HCC,in transforming growth factor-a (TGF-a) and c-Myc–TGF-atransgenic mice, characterized by different susceptibility toHCC, and in human HCC: (i) iNOS function and interactionswith nuclear factor-kB (NF-kB) and Ha-RAS/extracellularsignal-regulated kinase (ERK) during hepatocarcinogenesis;(ii) influence of genetic predisposition to liver cancer on thesepathways and role of these cascades in determining a susceptibleor resistant phenotype and (iii) iNOS prognostic value in humanHCC. We found progressive iNos induction in rat and mouse liverlesions, always at higher levels in the most aggressive models rep-resented by HCC of rats genetically susceptible to hepatocarcino-genesis and c-Myc–TGF-a transgenic mice. iNOS, inhibitor of kBkinase/NF-kB and RAS/ERK upregulation was significantly higherin HCC with poorer prognosis (as defined by patients' survivallength) and positively correlated with tumor proliferation, genomicinstability and microvascularization and negatively with apoptosis.Suppression of iNOS signaling by aminoguanidine led to decreasedHCC growth and NF-kB and RAS/ERK expression and increasedapoptosis both in vivo and in vitro. Conversely, block of NF-kBsignaling by sulfasalazine or short interfering RNA (siRNA) orERK signaling by UO126 caused iNOS downregulation in HCCcell lines. These findings indicate that iNOS cross talk with NF-kB and Ha-RAS/ERK cascades influences HCC growth and prog-nosis, suggesting that key component of iNOS signaling could rep-resent important therapeutic targets for human HCC.IntroductionHepatocellular carcinoma (HCC) is one of the most frequent anddeadliest human cancers worldwide. Current therapies do not improvesignificantly the prognosis of patients with unresectable HCC (1,2).This emphasizes the need to investigate the molecular mechanismsresponsible for HCC development to identify new targets for earlydiagnosis, chemoprevention and treatment.Numerous genes regulating susceptibility to HCC and controllinggrowth, progression and redifferentiation of preneoplastic and neo-plastic lesions have been mapped in rodents (3). Decrease in growthability and/or marked redifferentiation of preneoplastic lesion char-acterizes rodent strains resistant to hepatocarcinogenesis (3,4). Con-sequently, studies on the mechanisms underlying the acquisition ofa phenotype susceptible/resistantto hepatocarcinogenesis in rodentstrains, carrying preneoplastic lesions differently prone to progressto HCC, may lead to the discovery of prognostic markers and ther-apeutic targets for the human disease. Dysplastic nodules and HCCinduced in susceptible Fisher 344 (F344) rats show upregulation ofc-Myc, Cyclin D1, E and A and E2f1 genes, increased cyclinD1–Cdk4, cyclin E–Cdk2 and E2f1–Dp1 complexes and retinoblas-toma protein (pRb) hyperphosphorylation (4–6). These changes areabsent or less pronounced in liver lesions from resistant Brown Norway(BN) rats, where a block of