Impaired hepatitis C virus (HCV)-specific effector CD8+ T cells undergo massive apoptosis in the peripheral blood during acute HCV infection and in the liver during the chronic phase of infection

A majority of patients infected with hepatitis C virus (HCV) do not sustain an effective T-cell response, and viremia persists. The mechanism leading to failure of the HCV-specific CD8⁺ T-cell response in patients developing chronic infection is unclear. We investigated apoptosis susceptibility of HCV-specific CD8⁺ T cells during the acute and chronic stages of infection. Although HCV-specific CD8⁺ T cells in the blood during the acute phase of infection and in the liver during the chronic phase were highly activated and expressed an effector phenotype, the majority was undergoing apoptosis. In contrast, peripheral blood HCV-specific CD8⁺ T cells during the chronic phase expressed a resting memory phenotype. Apoptosis susceptibility of HCV-specific CD8⁺ T cells was associated with very high levels of programmed death-1 (PD-1) and low CD127 expression and with significant functional T-cell deficits. Further evaluation of the “death phase” of HCV-specific CD8⁺ T cells during acute HCV infection showed that the majority of cells were dying by a process of cytokine withdrawal, mediated by activated caspase 9. Contraction during the acute phase occurred rapidly via this process despite the persistence of the virus. Remarkably, in the chronic phase of HCV infection, at the site of infection in the liver, a substantial frequency of caspase 9-mediated T-cell death was also present. This study highlights the importance of cytokine deprivation-mediated apoptosis with consequent down-modulation of the immune response to HCV during acute and chronic infections.     

Acetylcholinesterases--the structural similarities and differences

Acetylcholinesterase (AChE) is a widely spread enzyme playing a very important role in nerve signal transmission. As AChE controls key processes, its inhibition leads to the very fast death of an organism, including humans. However, when this feature is to be used for killing of unwanted organisms (i.e. mosquitoes), one is faced with the question - how much do AChEs differ between species and what are the differences? Here, a theoretical point of view was utilized to identify the structural basis for such differences. The various primary and tertiary alignments show that AChEs are very evolutionary conserved enzymes and this fact could lead to difficulties, for example, in the search for inhibitors specific for a particular species.     

Mapping the organization of axis of motion selective features in human area MT using high-field fMRI

Functional magnetic resonance imaging (fMRI) at high magnetic fields has made it possible to investigate the columnar organization of the human brain in vivo with high degrees of accuracy and sensitivity. Until now, these results have been limited to the organization principles of early visual cortex (V1). While the middle temporal area (MT) has been the first identified extra-striate visual area shown to exhibit a columnar organization in monkeys, evidence of MT's columnar response properties and topographic layout in humans has remained elusive. Research using various approaches suggests similar response properties as in monkeys but failed to provide direct evidence for direction or axis of motion selectivity in human area MT. By combining state of the art pulse sequence design, high spatial resolution in all three dimensions (0.8 mm isotropic), optimized coil design, ultrahigh field magnets (7 Tesla) and novel high resolution cortical grid sampling analysis tools, we provide the first direct evidence for large-scale axis of motion selective feature organization in human area MT closely matching predictions from topographic columnar-level simulations.     

Measurement of photosynthesis by the dry weight increment of samples composed of leaf segments

V práci se popisuje zp?sob pou?ití metody váhového stanovení intensity fotosynthesy vzork? listového pletiva i pro rostliny s úzkými listovými ?epelemi (nap?. Festuca, Stipa aj.). Vzorek listového pletiva tvo?í zde piocha ráme?ku, který je vyplněn těsně vedle sebe le?ícími listovými úseky. Listové úseky dlouhé 26 mm se vysekávají z list? speciálním razidlem a jejich po?et ve vzorku je nutno pro ka?dý rostlinný druh p?edem experimentálně stanovit. V souvislosti s touto otázkou auto?i popisují závislosti hodnot intensity fotosynthesy na mno?ství úsek? ve vzorku. P?esnost nep?ímého stanovení su?iny kontrolními vzorky je p?i deseti stanoveních a pětiprocentní pravdépodobnosti maximálné ± 1 %, co? dává mo?nost p?i p?edpokládaném p?ír?stku su?iny o 10% mě?it intensitu fotosynthesy s p? esností na ±10%.     

Control of autoimmunity by "epitope theft"

We all possess T cells with autoaggressive potential. Knowledge of their regulation is crucial for elucidating pathogenetic pathways and designing effective treatments for autoimmune diseases. A novel mechanism of T-cell silencing--in an autoimmune model--has recently been identified and is termed "epitope theft". The "thieves" are naive CD8+ T cells, which apparently "steal" MHC-class-I-antigen complexes from antigen-presenting cells (APCs). The deprived APCs can no longer activate other potentially pathogenic naive CD8+ T cells that are specific for the same epitope. This phenomenon is a previously unrecognized antigen-specific mode of protection against autoimmunity.     

Involvement of the p97-Ufd1-Npl4 complex in the regulated endoplasmic reticulum-associated degradation of inositol 1,4,5-trisphosphate receptors

Inositol 1,4,5-trisphosphate (IP(3)) receptors form tetrameric, IP(3)-gated channels in endoplasmic reticulum membranes that govern the release of Ca(2+) from this organelle. In response to activation of certain G protein-coupled receptors that persistently elevate IP(3) concentration, IP(3) receptors are ubiquitinated and degraded by the ubiquitin-proteasome pathway. IP(3) receptor ubiquitination is mediated by the ubiquitin-conjugating enzyme, (mam)Ubc7, a component of the endoplasmic reticulum-associated degradation pathway. However, the mechanism by which ubiquitinated IP(3) receptors are transferred to the proteasome is not known. Here, we examine this process and show in several mammalian cell types that the ATPase p97 associates with IP(3) receptors in response to hormonal stimuli that induce IP(3) receptor ubiquitination. To examine the functional relevance of the p97 interaction with IP(3) receptors, we stably and specifically reduced p97 protein levels by 62 +/- 3% in Rat-1 fibroblasts using RNA interference. In these cells, endothelin-1-induced IP(3) receptor degradation was markedly retarded and the accumulation of ubiquitinated IP(3) receptors was markedly enhanced. These effects were reversed by expression of exogenous p97. In addition, Ufd1 and Npl4, which complex with p97, also associated with IP(3) receptors upon hormonal stimulation. We conclude that the p97-Ufd1-Npl4 complex couples ubiquitinated IP(3) receptors to proteasomal degradation and, thus, plays a key role in IP(3) receptor processing. These data also establish that the p97-Ufd1-Npl4 complex mediates endoplasmic reticulum-associated degradation in mammalian cells.     

First principles calculations of thermodynamics and kinetic parameters and molecular dynamics simulations of acetylcholinesterase reactivators: can mouse data provide new insights into humans?

We have applied a theoretical methodology, previously developed to evaluate the association and kinetic reactivation constants of oximes, comparing theoretical data obtained for human acetylcholinesterase (HsAChE) with in vitro results from Mus musculus AChE (MmAChE) previously reported in the literature. Our results, further checked by additional molecular dynamics simulations steps, showed a good correlation between the theoretical and experimental data, supporting the methodology as appropriate for prediction of thermodynamic and kinetic parameters and corroborated MmAChE as a suitable model for studies with HsAChE.     

Mycorrhizal responsiveness trends in annual crop plants and their wild relatives—a meta-analysis on studies from 1981 to 2010

Background and aims

Year of release of a cultivar reflects the agricultural and breeding practices of its time; we hypothesize that there are differences in mycorrhizal responsiveness of new high yielding and old crop plants and landraces. We evaluated the importance of the year of release on mycorrhizal responsiveness, arbuscular mycorrhizal (AM) fungal root colonization and P efficiency. We also analyzed the effect of experimental treatments, P efficiency (P acquisition and P utilization efficiency) and AM fungal root colonization on a potential mycorrhizal responsiveness trend for year of release.

Methods

We conducted a meta-analysis on 39 publications working on 320 different crop plant genotypes.

Results

New cultivars were less intensely colonized but were more mycorrhiza-responsive (and possibly dependent) compared to ancestral genotypes. This trend was potentially influenced by the moderator variables density, pre-germination, plant, plant type and AMF species. AM root colonization was also important for the mycorrhizal responsiveness trend for year of release, but P efficiency was not.

Conclusions

With the data available we could find no evidence that new crop plant genotypes lost their ability to respond to mycorrhiza due to agricultural and breeding practices.