Nutlin-3 overcomes arsenic trioxide resistance and tumor metastasis mediated by mutant p53 in Hepatocellular Carcinoma

Background: Arsenic trioxide has been demonstrated as an effective anti-cancer drug against leukemia and solid tumors both in vitro and in vivo. However, recent phase II trials demonstrated that single agent arsenic trioxide was poorly effective against hepatocellular carcinoma (HCC), which might be due to drug resistance. Methods: Mutation detection of p53 gene in arsenic trioxide resistant HCC cell lines was performed. The therapeutic effects of arsenic trioxide and Nutlin-3 on HCC were evaluated both in vitro and in vivo. A series of experiments including MTT, apoptosis assays, co-Immunoprecipitation, siRNA transfection, lentiviral infection, cell migration, invasion, and epithelial-mesenchy-mal transition (EMT) assays were performed to investigate the underlying mechanisms. Results: The acquisition of p53 mutation contributed to arsenic trioxide resistance and enhanced metastatic potential of HCC cells. Mutant p53 (Mutp53) silence could re-sensitize HCC resistant cells to arsenic trioxide and inhibit the metastatic activities, while mutp53 overexpression showed the opposite effects. Neither arsenic trioxide nor Nutlin-3 could exhibit obvious effects against arsenic trioxide resistant HCC cells, while combination of them showed significant effects. Nutlin-3 can not only increase the intracellular arsenicals through inhibition of p-gp but also promote the p73 activation and mutp53 degradation mediated by arsenic trioxide. In vivo experiments indicated that Nutlin-3 can potentiate the antitumor activities of arsenic trioxide in an orthotopic hepatic tumor model and inhibit the metastasis to lung. Conclusions: Acquisitions of p53 mutations contributed to the resistance of HCC to arsenic trioxide. Nutlin-3 could overcome arsenic trioxide resistance and inhibit tumor metastasis through p73 activation and promoting mutant p53 degradation mediated by arsenic trioxide

The role of autophagy in microbial infection and immunity

The autophagy pathway represents an evolutionarily conserved cell recycling process that is activated in response to nutrient deprivation and other stress signals. Over the years, it has been linked to an array of cellular functions. Equally, a wide range of cell-intrinsic, as well as extracellular, factors have been implicated in the induction of the autophagy pathway. Microbial infections represent one such factor that can not only activate autophagy through specific mechanisms but also manipulate the response to the invading microbe’s advantage. Moreover, in many cases, particularly among viruses, the pathway has been shown to be intricately involved in the replication cycle of the pathogen. Conversely, autophagy also plays a role in combating the infection process, both through direct destruction of the pathogen and as one of the key mediating factors in the host defense mechanisms of innate and adaptive immunity. Further, the pathway also plays a role in controlling the pathogenesis of infectious diseases by regulating inflammation. In this review, we discuss various interactions between pathogens and the cellular autophagic response and summarize the immunological functions of the autophagy pathway

Bayesian Hierarchical Modeling Monthly Crash Counts on Freeway Segments with Temporal Correlation

As the basis of traffic safety management, crash prediction models have long been a prominent focus in the field of freeway safety research. Studies usually take years or seasons as the observed time units, which may result in heterogeneity in crash frequency. To eliminate that heterogeneity, this study analyzes monthly crash counts and develops Bayesian hierarchical models with random effects, lag-1 autoregression (AR-1), and both (REAR-1) to accommodate the multilevel structure and temporal correlation in crash data. The candidate models are estimated and evaluated in the freeware WinBUGS using a crash dataset obtained from the Kaiyang Freeway in Guangdong Province, China. Significant temporal effects are found in the three models, and Deviance Information Criteria (DIC) results show that taking temporal correlation into account considerably improves the model fit compared with the Poisson model. The hierarchical models also avoid any misidentification of the factors with significant safety effects, because their variances are greater than in the Poisson model. The DIC value of the AR-1 model is substantially lower than that of the random effect model and equivalent to that of the REAR-1 model, which indicates the superiority of the lag-1 autoregressive structure in accounting for the temporal effects in crash frequency

Lack of the IFN-γ signal leads to lethal Orientia tsutsugamushi infection in mice with skin eschar lesions.

Scrub typhus is an acute febrile disease due to Orientia tsutsugamushi (Ot) infection and can be life-threatening with organ failure, hemorrhage, and fatality. Yet, little is known as to how the host reacts to Ot bacteria at early stages of infection; no reports have addressed the functional roles of type I versus type II interferon (IFN) responses in scrub typhus. In this study, we used comprehensive intradermal (i.d.) inoculation models and two clinically predominant Ot strains (Karp and Gilliam) to uncover early immune events. Karp infection induced sequential expression of Ifnb and Ifng in inflamed skin and draining lymph nodes at days 1 and 3 post-infection. Using double Ifnar1-/-Ifngr1-/- and Stat1-/- mice, we found that deficiency in IFN/STAT1 signaling resulted in lethal infection with profound pathology and skin eschar lesions, which resembled to human scrub typhus. Further analyses demonstrated that deficiency in IFN-γ, but not IFN-I, resulted in impaired NK cell and macrophage activation and uncontrolled bacterial growth and dissemination, leading to metabolic dysregulation, excessive inflammatory cell infiltration, and exacerbated tissue damage. NK cells were found to be the major cellular source of innate IFN-γ, contributing to the initial Ot control in the draining lymph nodes. In vitro studies with dendritic cell cultures revealed a superior antibacterial effect offered by IFN-γ than IFN-β. Comparative in vivo studies with Karp- and Gilliam-infection revealed a crucial role of IFN-γ signaling in protection against progression of eschar lesions and Ot infection lethality. Additionally, our i.d. mouse models of lethal infection with eschar lesions are promising tools for immunological study and vaccine development for scrub typhus

The carbonaceous aerosol levels still remain a challenge in the Beijing-Tianjin-Hebei region of China: Insights from continuous high temporal resolution measurements in multiple cities

Carbonaceous aerosols in high emission areas attract worldwide attention of the scientific community and the public due to their adverse impacts on the environment, human health and climate. However, long-term continuous hourly measurements are scarce on the regional scale. In this study, a one-year hourly measurement (from December 1, 2016 to November 30, 2017) of organic carbon (OC) and elemental carbon (EC) in airborne fine particles was performed using semi-continuous OC/EC analyzers in Beijing, Tianjin, Shijiazhuang and Tangshan in the Beijing-Tianjin-Hebei (BTH) region in China, which is one of high emission areas in China, even in the world. Marked spatiotemporal variations were observed. The highest concentrations of OC (22.8 ± 30.6 μg/m 3 ) and EC (5.4 ± 6.5 μg/m 3 ) occurred in Shijiangzhuang while the lowest concentrations of OC (11.0 ± 10.7 μg/m 3 ) and EC (3.1 ± 3.6 μg/m 3 ) were obtained in Beijing and Tianjin, respectively. Pronounced monthly, seasonal and diurnal variations of OC and EC were recorded. Compared to published data from the past two decades for the BTH region, our OC and EC levels were lower, implying some effect of recent measures for improving the air quality. Significant correlations of OC versus EC (p < 0.001) were found throughout the study period with high slopes and correlation coefficients in winter, but low slopes and correlation coefficients in summer. The estimated secondary OC (SOC), based on the minimum R squared (MRS) method, represented 29%, 47%, 38% and 48% of the OC for Beijing, Tianjin, Shijiazhuang and Tangshan, respectively. These percentages are larger than previous ones obtained for the BTH region in the past decade. There were obvious differences in the potential source regions of OC and EC among the four cities. Obvious prominent potential source areas of OC and EC were observed for Beijing, which were mainly located in the central and western areas of Inner Mongolia and even extended to the Mongolian regions, which is different from the findings in previous studies. For all sites, adjacent areas of the main provinces in northern China were found to be important potential source areas. © 2019 The Author

Steatosis and liver cancer in transgenic mice expressing the structural and nonstructural proteins of hepatitis C virus

Background and Aims: The aim of this study was to determine whether expression of hepatitis C virus proteins alters hepatic morphology or function in the absence of inflammation. Methods: Transgenic C57BL/6 mice with liver-specific expression of RNA encoding the complete viral polyprotein (FL-N transgene) or viral structural proteins (S-N transgene) were compared with nontransgenic littermates for altered liver morphology and function. Results: FL-N transcripts were detectable only by reverse-transcription polymerase chain reaction, and S-N transcripts were identified in Northern blots. The abundance of viral proteins was sufficient for detection only in S-N transgenic animals. There was no inflammation in transgenic livers, but mice expressing either transgene developed age-related hepatic steatosis that was more severe in males. Apoptotic or proliferating hepatocytes were not significantly increased. Hepatocellular adenoma or carcinoma developed in older male animals expressing either transgene, but their incidence reached statistical significance only in FL-N animals. Neither was ever observed in age-matched nontransgenic mice. Conclusions: Constitutive expression of viral proteins leads to common pathologic features of hepatitis C in the absence of specific anti-viral immune responses. Expression of the structural proteins enhances a low background of steatosis in C57BL/6 mice, while additional low level expression of nonstructural proteins increases the risk of cancer

Efficient assembly and annotation of the transcriptome of catfish by RNA-Seq analysis of a doubled haploid homozygote

BACKGROUND: Upon the completion of whole genome sequencing, thorough genome annotation that associates genome sequences with biological meanings is essential. Genome annotation depends on the availability of transcript information as well as orthology information. In teleost fish, genome annotation is seriously hindered by genome duplication. Because of gene duplications, one cannot establish orthologies simply by homology comparisons. Rather intense phylogenetic analysis or structural analysis of orthologies is required for the identification of genes. To conduct phylogenetic analysis and orthology analysis, full-length transcripts are essential. Generation of large numbers of full-length transcripts using traditional transcript sequencing is very difficult and extremely costly. RESULTS: In this work, we took advantage of a doubled haploid catfish, which has two sets of identical chromosomes and in theory there should be no allelic variations. As such, transcript sequences generated from next-generation sequencing can be favorably assembled into full-length transcripts. Deep sequencing of the doubled haploid channel catfish transcriptome was performed using Illumina HiSeq 2000 platform, yielding over 300 million high-quality trimmed reads totaling 27 Gbp. Assembly of these reads generated 370,798 non-redundant transcript-derived contigs. Functional annotation of the assembly allowed identification of 25,144 unique protein-encoding genes. A total of 2,659 unique genes were identified as putative duplicated genes in the catfish genome because the assembly of the corresponding transcripts harbored PSVs or MSVs (in the form of pseudo-SNPs in the assembly). Of the 25,144 contigs with unique protein hits, around 20,000 contigs matched 50% length of reference proteins, and over 14,000 transcripts were identified as full-length with complete open reading frames. The characterization of consensus sequences surrounding start codon and the stop codon confirmed the correct assembly of the full-length transcripts. CONCLUSIONS: The large set of transcripts assembled in this study is the most comprehensive set of genome resources ever developed from catfish, which will provide the much needed resources for functional genome research in catfish, serving as a reference transcriptome for genome annotation, analysis of gene duplication, gene family structures, and digital gene expression analysis. The putative set of duplicated genes provide a starting point for genome scale analysis of gene duplication in the catfish genome, and should be a valuable resource for comparative genome analysis, genome evolution, and genome function studies