Effects of oxidative and nitrative challenges on alpha-synuclein fibrillogenesis involve distinct mechanisms of protein modifications

Filamentous inclusions of alpha-synuclein protein are hallmarks of neurodegenerative diseases collectively known as synucleinopathies. Previous studies have shown that exposure to oxidative and nitrative species stabilizes alpha-synuclein filaments in vitro, and this stabilization may be due to dityrosine cross-linking. To test this hypothesis, we mutated tyrosine residues to phenylalanine and generated recombinant wild type and mutant alpha-synuclein proteins. alpha-Synuclein proteins lacking some or all tyrosine residues form fibrils to the same extent as the wild type protein. Tyrosine residues are not required for protein cross-linking or filament stabilization resulting from transition metal-mediated oxidation, because higher Mr SDS-resistant oligomers and filaments stable to chaotropic agents are detected using all Tyr --> Phe alpha-synuclein mutants. By contrast, cross-linking resulting from exposure to nitrating agents required the presence of one or more tyrosine residues. Furthermore, tyrosine cross-linking is involved in filament stabilization, because nitrating agent-exposed assembled wild type, but not mutant alpha-synuclein lacking all tyrosine residues, was stable to chaotropic treatment. In addition, the formation of stable alpha-synuclein inclusions in intact cells after exposure to oxidizing and nitrating species requires tyrosine residues. These findings demonstrate that nitrative and/or oxidative stress results in distinct mechanisms of alpha-synuclein protein modifications that can influence the formation of stable alpha-synuclein fibrils.     

Cdc45 Is a Critical Effector of Myc-Dependent DNA Replication Stress

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The human placental derived BeWo cell line: a useful model for selecting Plasmodium falciparum CSA-binding parasites

Chondroitin sulfate A (CSA) present in the placental intervillous blood spaces has been described as the main receptor involved in the massive sequestration of Plasmodium falciparum parasitized erythrocytes to the placenta. Placental parasite isolates are functionally distinct from isolates that sequester in other organs, because they do not cytoadhere to CD36 but instead bind to CSA. To investigate for the parasites molecules associated with the CSA adhesion phenotype, different methodologies have been developed to select for CSA-binding lines in vitro mainly using non-placental sources of CSA that differ in their sulfation pattern. In this study, we show that the human trophoblastic BeWo cell line is a very efficient alternative to select for the CSA-binding phenotype in parasitized erythrocytes.     

Are biodiversity patterns of saproxylic beetles shaped by habitat limitation or dispersal limitation? A case study in unfragmented montane forests

Understanding the processes that shape biodiversity patterns is essential for ecosystem management and conservation. Local environmental conditions are often good predictors of species distribution and variations in habitat quality usually positively correlate to species richness. However, beside habitat limitation, species presence-absence may be constrained by dispersal limitation. We tested the relative importance of both limitations on saproxylic beetle diversity, using forest continuity as a surrogate for dispersal limitation and stand maturity as a surrogate for habitat limitation. Forest continuity relies on the maintenance of a forest cover over time, while stand maturity results in the presence of old-growth habitat features. Forty montane beech-fir forests in the French pre-Alps were sampled, under a balanced sampling design in which forest continuity and stand maturity were crossed. A total of 307 saproxylic beetle species were captured using flight-interception traps and Winkler–Berlese extractors. We explored the response of low- versus high-dispersal species groups to forest continuity and stand maturity. Saproxylic beetle diversity increased significantly with stand maturity and was mostly influenced by variables related to deadwood diversity at the stand scale and suitable habitat availability at the landscape scale. Surprisingly, no evidence of dispersal limitation was found, as diversity patterns were not influenced by forest continuity and associated variables, even for low-dispersal species. Our study demonstrates that in an unfragmented forest landscape, saproxylic beetles are able to colonize recent forests, as long as local deadwood resources are sufficiently diversified (e.g. tree species, position, diameter and/or decay stage).     

Species differences in the generation of the chiral sulfoxide metabolite of albendazole in sheep and rats

The prochiral anthelmintic drug albendazole was administered orally to sheep and rats. Blood samples were taken at standardized intervals during the time course of the plasma kinetics: 18 h in rats and 48 h in sheep. The enantiomeric ratio of the sulfoxide metabolite was determined by means of HPLC on a chiral stationary phase, the chiral selector of which was a N-3,5-dinitrobenzoyl derivative of (S)-tyrosine. Two enantiomers were detected in both animal species but their ratios were inverted in rat vs. sheep. The evolution of the ratio is turned from a racemate at 15 min to 60(-):40(+) at 12 h in rats, while it moved from 23(-):77(+) at 3 h to 4(-):96(+) at 36 h after administration in sheep.     

Adaptations for the maintenance of water balance by three species of Antarctic mites

Three species of Antarctic mites, Alaskozetes antarcticus, Hydrogamasellus antarcticus and Rhagidia gerlachei, are abundant in the vicinity of Palmer Station, Antarctica. No single mechanism for reducing water stress was shared by all three species. A. antarcticus and R. gerlachei (both ca. 200 μg) are over twice as large as H. antarcticus (ca. 90 μg), but all had similar body water content (67%) and tolerated a loss of up to 35% of their body water before succumbing to dehydration. All imbibed free water and had the capacity to reduce water loss behaviorally by forming clusters. Alaskozetes antarcticus was distinct in that it relied heavily on water conservation (xerophilic classification) that was largely achieved by its thick cuticular armor, a feature shared by all members of this suborder (Oribatida), and abundant cuticular hydrocarbons. In comparison to the other two species, A. antarcticus was coated with 2–3× the amount of cuticular hydrocarbons, had a 20-fold reduction in net transpiration rate, and had a critical transition temperature (CTT) that indicates a pronounced suppression in activation energy (E _a) at temperatures below 25°C. In contrast, H. antarcticus and R. gerlachei lack a CTT, have lower amounts of cuticular hydrocarbons and have low E _as and high net transpiration rates, classifying them as hydrophilic. Only H. antarcticus was capable of utilizing water vapor to replenish its water stores, but it could do so only at relative humidities close to saturation (95–98 %RH). Thus, H. antarcticus and R. gerlachei require wet habitats and low temperature to counter water loss, and replace lost water behaviorally through predation. Compared to mites from the temperate zone, all three Antarctic species had a lower water content, a feature that commonly enhances cold tolerance.