Protein kinase C is required for the disappearance of MPF upon artificial activation in mouse eggs

The aim of the present study was to investigate the implication of protein kinase C (PKC) in the mouse egg activation process. We used OAG (1-oleoyl-2-acetyl-sn-glycerol) as a PKC activator, calphostin C as a specific PKC inhibitor, and the calcium ionophore A23187 as a standard parthenogenetic agent. The exposure of zona-free eggs to 150 microM or 50 microM OAG for 10 min resulted in meiosis II completion in approximately 80% of instances. By contrast, at a lower concentration (25 microM), the PKC stimulator was ineffective as parthenogenetic agent. Shortly after the application of 150 microM OAG, the cytosolic Ca2+ concentration ([Ca2+]i) increased transiently in all the eggs examined, whereas after the addition of 50 microM OAG, [Ca2+]i remained unchanged for at least 20 min. During this period, the activity of M-phase promoting factor (MPF) dramatically decreased and most of the eggs entered anaphase except when the PKC was inhibited by calphostin C. Similarly, MPF inactivation and meiosis resumption were prevented in calphostin C-loaded eggs following treatment with A23187, even though the ionophore-induced Ca2+ signalling was not affected. Taken together, our results indicate that stimulation of PKC is a sufficient and necessary event to induce meiosis resumption in mouse eggs and strongly suggest that, in this species, the mechanism by which a transient calcium burst triggers MPF inactivation involves a PKC-dependent pathway.     

The impacts of road and walking trails upon adjacent vegetation: effects of road building materials on species composition in a nutrient poor environment

Roads represent an important landscape element affecting both biotic and abiotic components. Alteration of soil properties along roads (addition of nutrients) is assumed to have a great impact on vegetation structure especially in nutrient poor ecosystems. Existing studies focus mainly on road dust. In our study we assessed the overall effects of roads upon adjacent alpine tundra vegetation and soils in Krkonoše Mts, Czech Republic. Our aims were to (1) reconstruct the road-related changes using aerial data and GPS mapping to study colonization of roadside plant species; (2) assess the road effects on physical-chemical soil properties and vegetation composition along transects; and (3) propose conservation measures to stop further damage. Changes were reconstructed from historical multispectral aerial photography (1986 to 1997), measured by GPS device (1997, 2004), and accompanied by detailed soil (1998, 2000 and 2001) and vegetation (2000 and 2004) surveys along transects. Along alkaline roads, fast and profound shifts in physical-chemical soil properties (pH increased from 3.9 up to 7.6, base saturation from 9-30% up to 100%), and species composition were recorded. The roadside vegetation doubled in area during the studied decade. Stress-tolerant tundra species were replaced by meso- to nitrophilous species and species preferring man-made habitats. The intensity of changes depended significantly on the type of road material and the position relative to the road (slope position, distance from the road). Our findings support the assumption that alkaline gravel is the main cause of changes along roads in the area, and indicate the leading role of water transport in the soil and consequent vegetation alteration. To prevent the further damage we recommended replacement of alkaline gravel by granite, even though expensive and technically complicated. Based on our recommendations, the National Park authorities started to reconstruct the trails, although recovery is expected to be slow.     

Energy policy scenarios to reach challenging climate protection targets in the German electricity sector until 2050

In this article we demonstrate how challenging greenhouse gas reduction targets of up to 95% until 2050 can be achieved in the German electricity sector. In the analysis, we focus on the main requirements to reach such challenging targets. To account for interdependencies between the electricity market and the rest of the economy, different models were used to account for feedback loops with all other sectors. We include scenarios with different runtimes and retrofit costs for existing nuclear plants to determine the effects of a prolongation of nuclear power plants in Germany. Key findings for the electricity sector include the importance of a European-wide coordinated electricity grid extension and the exploitation of regional comparative cost effects for renewable sites. Due to political restrictions, nuclear energy will not be available in Germany in 2050. However, the nuclear life-time extension has a positive impact on end consumer electricity prices as well as economic growth in the medium term, if retrofit costs do not exceed certain limits.     

Autogenous healing and reinforcement corrosion of water-penetrated separation cracks in reinforced concrete

Depending on the crack width, the thickness of the structure, the water pressure, and the degree of acid of the water, long-term investigations have been performed over a period of 2 years with respect to the autogenous healing and reinforcement corrosion of water-penetrated separation cracks in reinforced concrete by the University of Kaiserslautern, supported by the Deutsche Forschungsgemeinschaft (DFG). For the waters penetrating the cracks deionised water (neutral, pH=7.0), and boric acid treated deionised water with a pH-value of 6.1 and 5.2 (weakly acid waters) were used. A complete autogenous healing could not be observed. The water penetrating the cracks could hardly be measured with a pH-value of 7.0 at the end of the test. While naturally at the beginning of the test, no influence of the water-chemical degree of the acids could be determined, the existing flow-through quantities towards the end of the test period depended clearly on the crack width and the pH-value. With an increasing crack width and an increasing acid-degree larger flow-through quantities were measured. Depending on the pH-value and the crack width it was determined whether and to which extent corrosion developed at the reinforcing steel bars crossing the cracks. With a crack width of 0.1 mm, corrosion was not to be observed in any case. For the test specimens with a crack width of 0.2 mm a start of the corrosion was found depending on the pH-value. With an increasing width of the crack, an increasing corrosion development is to be expected for test specimens penetrated by acid water. For a crack width of 0.4 mm and a pH-value of 5.2, the highest corrosion development was to be observed, however, there were weakenings of the cross section not worth being mentioned even after a 2-year test period.     

Amyloid Beta-Mediated Changes in Synaptic Function and Spine Number of Neocortical Neurons Depend on NMDA Receptors

Onset and progression of Alzheimer’s disease (AD) pathophysiology differs between brain regions. The neocortex, for example, is a brain region that is affected very early during AD. NMDA receptors (NMDARs) are involved in mediating amyloid beta (Aβ) toxicity. NMDAR expression, on the other hand, can be affected by Aβ. We tested whether the high vulnerability of neocortical neurons for Aβ-toxicity may result from specific NMDAR expression profiles or from a particular regulation of NMDAR expression by Aβ. Electrophysiological analyses suggested that pyramidal cells of 6-months-old wildtype mice express mostly GluN1/GluN2A NMDARs. While synaptic NMDAR-mediated currents are unaltered in 5xFAD mice, extrasynaptic NMDARs seem to contain GluN1/GluN2A and GluN1/GluN2A/GluN2B. We used conditional GluN1 and GluN2B knockout mice to investigate whether NMDARs contribute to Aβ-toxicity. Spine number was decreased in pyramidal cells of 5xFAD mice and increased in neurons with 3-week virus-mediated Aβ-overexpression. NMDARs were required for both Aβ-mediated changes in spine number and functional synapses. Thus, our study gives novel insights into the Aβ-mediated regulation of NMDAR expression and the role of NMDARs in Aβ pathophysiology in the somatosensory cortex.     

Modelling of HLA-DQ2 and its interaction with gluten peptides to explain molecular recognition in celiac disease

Celiac disease (CD) is sustained by abnormal intestinal mucosal T-cell response to gluten and it is strongly associated with HLA class II molecules encoded by DQA1*0501/DQB1*02 (DQ2) or DQA1*03/DQB1*0302 (DQ8). The in vitro stimulatory activity of gliadin increases after treatment with tissue transglutaminase (tTG) which catalyses the deamidation of specific residues of glutamine to glutamate that can serve as anchors for binding to DQ2 as well as to DQ8 molecules. We modelled the three-dimensional structure of the DQ2 dimer protein, the most frequent in celiac patients, by using a homology modelling strategy, and deposited the model in the Protein Data Bank (PDB). Then, we simulated the interactions of DQ2 with different gluten peptides and the deamidation of specific peptide glutamines in the known p4, p6, p7 and p9 anchor positions, as well as in p1 and p5 positions, and other substitutions for which experimental effects on binding are available by previous experimental studies. By evaluating the energy of interaction and the H-bond interactions, we were able to distinguish what substitutions improve the interaction peptide-DQ2, in agreement with previously published experimental data. By analysing the peptide-DQ2 complex at the atom level, we observed that these glutamate side chains can interact with specific positively charged amino acids of DQ2, absent in other HLA alleles not related to celiac disease. The simulation was also extended to other peptides, related to celiac disease but for which no experimental data exists about the effects of glutamine deamidation. Our results give an interpretation at the molecular level of previously reported binding experimental data and open a new window to gain further insights about peptide recognition in celiac disease.