Efficient, trans-complementing packaging systems for chimeric, pseudoinfectious dengue 2/yellow fever viruses

In our previous studies, we have stated to build a new strategy for developing defective, pseudoinfectious flaviviruses (PIVs) and applying them as a new type of vaccine candidates. PIVs combined the efficiency of live vaccines with the safety of inactivated or subunit vaccines. The results of the present work demonstrate further development of chimeric PIVs encoding dengue virus 2 (DEN2V) glycoproteins and yellow fever virus (YFV)-derived replicative machinery as potential vaccine candidates. The newly designed PIVs have synergistically functioning mutations in the prM and NS2A proteins, which abolish processing of the latter proteins and make the defective viruses capable of producing either only noninfectious, immature and/or subviral DEN2V particles. The PIV genomes can be packaged to high titers into infectious virions in vitro using the NS1-deficient YFV helper RNAs, and both PIVs and helpers can then be passaged as two-component genome viruses at an escalating scale.     

TP53 Gene Mutations Are Rare in Nondysplastic Barrett's Esophagus

In search of potential prognostic markers, we analyzed a large series of tissues of Barrett's esophagus and samples of adenocarcinomas arising in the terrain of Barrett's esophagus for TP53 gene mutations by direct sequencing of exons 5 to 9 of the TP53 gene. While 9 of 21 adenocarcinomas tested (42.9%) contained a TP53 mutation, none of 24 samples from Barrett's esophagus were mutated. This observation suggests that TP53 gene mutation may be a relatively late event in the progression from nondysplastic Barrett's esophagus to adenocarcinoma of esophagus. Therefore, TP53 gene mutations alone are not likely to represent a widely useful prognostic marker of the risk of progression to malignancy, at least not in Barrett's esophagus without dysplasia. This work was supported by grant ND/7009-3 from the Internal Grant Agency of the Ministry of Health of the Czech Republic.     

Albumin reduces thrombogenic potential of single-walled carbon nanotubes

Reduction of thrombogenicity of carbon nanotubes is an important prerequisite for their biomedical use. We assessed the thrombogenic activity of carboxylated single-walled carbon nanotubes (c-SWCNTs) and covalently PEGylated c-SWNCTs (PEG-SWCNTs) by testing the clotting time of platelet poor plasma and platelet aggregation in whole blood samples, and evaluated the impact of human serum albumin on thrombogenicity of carbon nanotubes. Both types of SWCNTs exhibited considerable thrombogenic activity. SWCNTs accelerated plasma clotting, with a lesser effect seen for PEG-SWCNTs. Treatment of SWCNTs with albumin did not affect the SWCNT-induced shortening of clotting time. In whole blood, no discernible differences in the effect of c-SWCNTs and PEG-SWCNTs on platelets were observed. Upon addition of SWCNTs to blood, dose- and time-dependent formation of agglomerates of nanotubes and platelets was demonstrated. Pretreatment of SWCNTs with albumin reduced the platelet aggregation: the number of single platelets increased, and the size of platelet-SWCNT agglomerates decreased dramatically. Hence, addition of albumin may serve to attenuate the adverse, thrombogenic effect of CNTs.     

Phosphoprotein associated with glycosphingolipid-enriched microdomains/Csk-binding protein: a protein that matters

Engagement of plasma membrane receptors is followed by an assembly of a multimolecular complex of signaling molecules organized by scaffolding or adaptor proteins. PAG is a recently characterized transmembrane adaptor protein associated with lipid rafts, which is involved in the regulation of Src-kinases, monomeric Ras protein, and interactions with the cytoskeleton. The review provides up-to-date information about the protein that attracts increasing attention in current biomedical research.     

Molecular and phenotypic characterization of Acinetobacter strains able to degrade diesel fuel

Characterization of bacterial communities in oil-contaminated soils and evaluation of their degradation capacities may serve as a guide for improving remediation of such environments. Using physiological and molecular methods, the aim of this work was to characterize 17 Acinetobacter strains (13 species) able to use diesel fuel oil as sole carbon and energy source. The strains were first tested for their ability to grow on different alkanes on minimal medium containing high NaCl concentrations. The envelope hydrophobicity of each strain was assessed by microbial adhesion to the hydrocarbon test (MATH) when grown in LB medium or minimal medium containing succinate or diesel fuel. Most strains were hydrophobic both in LB and minimal medium, except for strain Acinetobacter venetianus VE-C3 that was hydrophobic only in minimal medium. Furthermore, two A. venetianus strains, RAG-1(T) and LUH 7437, and strain ATCC 17905 (genomic species 13BJ) displayed biosurfactant activity. The alkM gene encoding alkane hydroxylase was detected in the chromosome of the 15 strains by PCR amplification, sequencing and Southern blot analysis. Phenotype microarray analysis performed on the five A. venetianus strains revealed that they differentially used purines as N-source and confirmed that they are unable to use carbohydrates.     

MiR-429 is linked to metastasis and poor prognosis in renal cell carcinoma by affecting epithelial-mesenchymal transition

Tumor Biology - MicroRNAs (miRNAs) have been proven to be important oncogenes and tumor suppressors in wide range of cancers, including renal cell carcinoma (RCC). In our study, we evaluated...     

Consistent blind protein structure generation from NMR chemical shift data

Protein NMR chemical shifts are highly sensitive to local structure. A robust protocol is described that exploits this relation for de novo protein structure generation, using as input experimental parameters the (13)C(alpha), (13)C(beta), (13)C', (15)N, (1)H(alpha) and (1)H(N) NMR chemical shifts. These shifts are generally available at the early stage of the traditional NMR structure determination process, before the collection and analysis of structural restraints. The chemical shift based structure determination protocol uses an empirically optimized procedure to select protein fragments from the Protein Data Bank, in conjunction with the standard ROSETTA Monte Carlo assembly and relaxation methods. Evaluation of 16 proteins, varying in size from 56 to 129 residues, yielded full-atom models that have 0.7-1.8 A root mean square deviations for the backbone atoms relative to the experimentally determined x-ray or NMR structures. The strategy also has been successfully applied in a blind manner to nine protein targets with molecular masses up to 15.4 kDa, whose conventional NMR structure determination was conducted in parallel by the Northeast Structural Genomics Consortium. This protocol potentially provides a new direction for high-throughput NMR structure determination.     

Comparison of Different Treatment Modalities Outcomes in Clinically Node-positive Bladder Cancer: Analysis of a Population-based Cancer Registry

IntroductionPatients with clinically node-positive bladder cancer were historically considered to have uniformly poor prognosis and were frequently treated with palliative chemotherapy (CHT) only. Although retrospective data show that long-term survival with combined treatment (surgery + CHT) is possible in one-third of these patients, consensus on a treatment algorithm is still lacking. The aim of the study is to compare the efficacy of different treatment modalities based on data from a population-based cancer registry.Patients and MethodsThe study comprises 661 patients identified from the Czech National Cancer Registry (1996-2015) with cTanyN1-3M0 bladder cancer; 195 were treated with CHT alone, 234 underwent radical cystectomy alone (RC), and 232 received a combination of RC and perioperative CHT (RC + CHT). Multivariate Cox proportional hazard regression analyses were used to evaluate the effectiveness of various treatments.ResultsThe 5-year OS for CHT alone, RC alone, and RC + CHT were 21.7% (95% confidence interval [CI], 15.4%-28.0%), 12.1% (95% CI, 7.4%-16.7%), and 25.4% (95% CI, 18.9%-31.9%), respectively (P < .001). The median survivals were 17, 10, and 23 months, respectively. In multivariate analysis, age > 60 years (hazard ratio, 1.29; 95% CI, 1.06-1.56; P = .011) and clinical stage cT3-4 (hazard ratio, 1.39; 95% CI, 1.12-1.71; P = .002) were negative predictors of survival. When compared with CHT, RC + CHT reduced the risk of overall mortality by 21% (P = .044).ConclusionApproximately one-quarter of clinically node-positive patients may achieve long-term survival with combined treatment integrating RC and perioperative CHT. The overall survival of patients is significantly improved with a multimodal approach in comparison to CHT alone.