Use of Non-Invasive Parameters of Non-Alcoholic Steatohepatitis and Liver Fibrosis in Daily Practice - An Exploratory Case-Control Study

Background

Non-alcoholic fatty liver disease (NAFLD) is the hepatic manifestation of a metabolic syndrome. To date, liver biopsy has been the gold standard used to differentiate between simple steatosis and steatohepatitis/fibrosis. Our aim was to compare the relevance of serum non-invasive parameters and scoring systems in the staging of liver fibrosis and non-alcoholic steatohepatitis (NASH) in patients with NAFLD.

Methods and Findings

A total of 112 consecutive patients diagnosed with NAFLD were included. A liver biopsy was performed on 56 patients. The Kleiner score was used for the staging and grading of the histology. Non-invasive parameters for fibrosis (hyaluronic acid; AST/ALT; fibrosis scoring indexes OELF, ELF, BARD score, APRI, NAFLD fibrosis score); and inflammation (M30 and M65 cytokeratin-18 fragments) were measured and calculated. The same analyses were performed in 56 patients diagnosed with NAFLD, who were not indicated for liver biopsy. Based on the liver histology, NASH was diagnosed in 38 patients; simple steatosis in 18 patients. A cut-off value of 750 U/L of serum M65 discriminated patients with and without NASH with a 80% sensitivity and 82% specificity (95% CI:57–95). Fibrosis stage F0–F2 was present in 39 patients; F3–F4 in 17 patients. Serum concentrations of hyaluronic acid were higher in patients with advanced fibrosis (p<0.01); a cut-off value of 25 µg/l discriminated patients with F3–F4 with a 90% sensitivity and 84% specificity from those with F0–F2 (95% CI:59–99). When applying the non-invasive criteria to those patients without a liver biopsy, NASH could only be diagnosed in 16%; however, advanced fibrosis could be diagnosed in 35% of them.

Conclusions

In patients with NAFLD, non-invasive serum parameters with a high accuracy can differentiate those patients with NASH and/or advanced fibrosis from those with simple steatosis. A substantial portion of those patients not indicated for liver biopsy might have undiagnosed advanced fibrosis.     

Evolution of Tertiary Structure of Viral RNA Dependent Polymerases

Viral RNA dependent polymerases (vRdPs) are present in all RNA viruses; unfortunately, their sequence similarity is too low for phylogenetic studies. Nevertheless, vRdP protein structures are remarkably conserved. In this study, we used the structural similarity of vRdPs to reconstruct their evolutionary history. The major strength of this work is in unifying sequence and structural data into a single quantitative phylogenetic analysis, using powerful a Bayesian approach.The resulting phylogram of vRdPs demonstrates that RNA-dependent DNA polymerases (RdDPs) of viruses within Retroviridae family cluster in a clearly separated group of vRdPs, while RNA-dependent RNA polymerases (RdRPs) of dsRNA and +ssRNA viruses are mixed together. This evidence supports the hypothesis that RdRPs replicating +ssRNA viruses evolved multiple times from RdRPs replicating +dsRNA viruses, and vice versa. Moreover, our phylogram may be presented as a scheme for RNA virus evolution. The results are in concordance with the actual concept of RNA virus evolution. Finally, the methods used in our work provide a new direction for studying ancient virus evolution.     

Removable nanocoatings for siRNA polyplexes

To assist in overcoming the inherent instability of nucleic acid-containing polyplexes in physiological solutions, we have here set out to develop removable nanocoatings for modifying the surface of siRNA-based nanoparticles. Here, N-(2-hydroxypropyl)methacrylamide (HPMA) based copolymers containing carbonylthiazolidine-2-thione (TT) reactive groups in their side chains bound via disulfide spacers to the polymeric backbone were synthesized, and these copolymers were used to coat the surface of polyplexes formed by the self-assembly of anti-Luciferase siRNA with the polycations polyethylene imine (PEI) and poly(HPMA)-grafted poly(l-lysine) (GPL). The coating process was monitored by analyzing changes in the weight-average molecular weight (M(w)), the hydrodynamic radius (R(h)), and the zeta-potential (ζ) of the polyplexes, using both static (SLS) and dynamic (DLS) light scattering methods. The outlined methods resulted in the attachment of, on average, 28 polymer molecules to the surface of the polyplexes, forming a ～5-nm-thick hydrophilic stealth coating. Initial efforts to develop RGD-targeted coated polyplexes are also described. Atomic force microscopy (AFM) showed an angular polyplex structure and confirmed the narrow size distribution of the coated nanoparticles. The stability of the polymer-coated and uncoated polyplexes was evaluated by gel electrophoresis and by turbidity measurements, and it was found that modifying the surface of the siRNA-containing polyplexes substantially improved their stability in physiological solutions. Using polymer-coated GPL-based polyplexes containing anti-Luciferase siRNA, we finally also obtained some initial proof-of-principle for time- and concentration-dependent target-specific gene silencing, suggesting that these systems hold significant potential for further in vitro and in vivo evaluation.     

A novel application of capnography during controlled human exposure to air pollution

Background  

The objective was to determine the repeatability and stability of capnography interfaced with human exposure facility.     

Morphological and molecular characterization of the Armillaria cepistipes – A. gallica complex in the Czech Republic and Slovakia

Armillaria cepistipes and A. gallica (Basidiomycota, Physalacriaceae) are morphologically similar species, and they are often nearly indistinguishable using DNA-based methods targeting the ITS region of ribosomal DNA. The aim of this study was to examine morphological and ecological features of A. cepistipes and A. gallica, and to test other DNA-based methods to distinguish the two species. Our results revealed discriminative macro- and micromorphological features between these two species, especially the presence of a distinct central pileus ocella, the shape of the annulus, the character of the velar stipe remnants and the length of the terminal cells of the pileus scales. Ecologically, A. gallica generally prefers warmer areas in lowlands (oak and alluvial forests), while A. cepistipes is more common in hilly and lower montane beech forests in Central Europe. Nevertheless, despite differences in ecological preferences, certain locations between 300 and 500 m a.s.l. are known to sympatrically support both species. The sequences of the translation elongation factor 1-alpha showed high interspecific variability, and this gene is a more appropriate candidate for distinguishing A. gallica from A. cepistipes.     

Exposure to fenitrothion causes malfunctions of Spodoptera exigua Hübn. (Lep., Noctuidae) eggs

A long-term exposition of Spodoptera exigua (Hübner) larvae to fenitrothion caused abnormalities in egg structure. When examined under a scanning electron microscope, the eggs revealed diminutions and cracks around the micropylar and aeropylar region. The damage was proportional to the concentration of the insecticide. The observed changes may be one of the reasons for the decreased survival of populations exposed to fenitrothion.     

Holocene dynamics of the alpine timberline in the High Sudetes

The study focuses on the High Sudetes that represent the most distinctive islands of alpine forest-free area among hercynian mid-mountains of Central Europe. Based on data from newly taken cores and previously published pollen profiles, comparison of the development of the alpine timberline position is carried out. The first of the analysed pollen profiles — the Labsky důl core in the Krkonoše Mts spans the whole period of the Holocene, the Keprník and Mezikotlí profiles in the Hruby Jeseník Mts bring information from the Subboreal/older Subatlanticum turn to the present. An exceptional position of the Krkonoše Mts in terms of permanent presence of the alpine belt throughout the Holocene was confirmed. Three oscillations of the alpine timberline during the Lower Holocene were detected in the profile from the Labsky důl site. In the Hruby Jeseník Mts a temperature dependent forest-free area existed at least since Subboreal to the present.