Chemical strategies for controlling protein folding and elucidating the molecular mechanisms of amyloid formation and toxicity

It has been more than a century since the first evidence linking the process of amyloid formation to the pathogenesis of Alzheimer's disease. During the last three decades in particular, increasing evidence from various sources (pathology, genetics, cell culture studies, biochemistry, and biophysics) continues to point to a central role for the pathogenesis of several incurable neurodegenerative and systemic diseases. This is in part driven by our improved understanding of the molecular mechanisms of protein misfolding and aggregation and the structural properties of the different aggregates in the amyloid pathway and the emergence of new tools and experimental approaches that permit better characterization of amyloid formation in vivo. Despite these advances, detailed mechanistic understanding of protein aggregation and amyloid formation in vitro and in vivo presents several challenges that remain to be addressed and several fundamental questions about the molecular and structural determinants of amyloid formation and toxicity and the mechanisms of amyloid-induced toxicity remain unanswered. To address this knowledge gap and technical challenges, there is a critical need for developing novel tools and experimental approaches that will not only permit the detection and monitoring of molecular events that underlie this process but also allow for the manipulation of these events in a spatial and temporal fashion both in and out of the cell. This review is primarily dedicated in highlighting recent results that illustrate how advances in chemistry and chemical biology have been and can be used to address some of the questions and technical challenges mentioned above. We believe that combining recent advances in the development of new fluorescent probes, imaging tools that enabled the visualization and tracking of molecular events with advances in organic synthesis, and novel approaches for protein synthesis and engineering provide unique opportunities to gain a molecular-level understanding of the process of amyloid formation. We hope that this review will stimulate further research in this area and catalyze increased collaboration at the interface of chemistry and biology to decipher the mechanisms and roles of protein folding, misfolding, and aggregation in health and disease.     

In vivo induction of heat shock proteins in the substantia nigra following L-DOPA administration is associated with increased activity of mitochondrial complex I and nitrosative stress in rats: regulation by glutathione redox state

Increasing evidence suggests a critical role for oxidative and nitrosative stress in the pathogenesis of most important neurodegenerative disorders. Parkinson's disease (PD) is a neurodegenerative disease characterized by a severe depletion in number of dopaminergic cells of the substantia nigra (SN). Administration of L-DOPA (LD) is the more effective treatment for patients with PD. However, the vast majority of patients suffer LD-related complications, which represent the major problem in the clinical management of PD. In the present study, LD administration to rats resulted in a significant dose-dependent increase in Hsp70 synthesis which was specific for the SN. The amount of 70 kDa protein increased after 6 h treatment reaching the maximal induction after 24-48 h. Induction of Hsp70 in the SN was associated with a significant increase in constitutive Hsc70 and mitochondrial Hsp60 stress proteins, and with increased expression of mitochondrial complex I whereas no significant changes were found in the activity of complex IV. In the same experimental conditions, a significant decrease in reduced glutathione was observed, which was associated with an increased content of oxidized glutathione content as well as nitric oxide (NO) synthase activity, NO metabolites and nitrotyrosine immunoreactivity. Interestingly, Hsp70 induction, iNOS up-regulation and nitrotyrosine formation have been confirmed also in SN and striatum of rats treated with LD and carbidopa, this latter being an inhibitor of the peripheral DOPA decarboxylase. Our data are in favor of the importance of the heat shock signal pathway as a basic mechanism of defense against neurotoxicity elicited by free radical oxygen and nitrogen species produced in aging and neurodegenerative disorders.     

Long‐term trends in restoration and associated land treatments in the southwestern United States

Restoration treatments, such as revegetation with seeding or invasive species removal, have been applied on U.S. public lands for decades. Temporal trends in these management actions have not been extensively summarized previously, particularly in the southwestern United States where invasive plant species, drought, and fire have altered dryland ecosystems. We assessed long‐term (1940–2010) trends in restoration using approximately 4,000 vegetation treatments conducted on Bureau of Land Management lands across the southwestern United States. We found that since 1940, the proportions of seeding and vegetation/soil manipulation (e.g. vegetation removal or plowing) treatments have declined, while the proportions of prescribed burn and invasive species treatments have increased. Treatments in pinyon‐juniper and big sagebrush communities declined in comparison to treatments in desert scrub, creosote bush, and riparian woodland communities. Restoration‐focused treatment objectives increased relative to resource extraction objectives. Species richness and proportion of native species used in seeding treatments also increased. Inflation‐adjusted costs per area rose 750% for vegetation/soil manipulation, 600% for seeding, and 400% for prescribed burn treatments in the decades from 1981 to 2010. Seeding treatments were implemented in warmer and drier years when compared to the climate conditions of the entire study period and warmer and wetter years relative to several years before and after the treatment. These results suggest that treatments over a 70‐year period on public lands in the southwestern United States are shifting toward restoration practices that are increasingly large, expensive, and related to fire and invasive species control.     

The spider communities on peat and upland grasslands In northern England

Between midsummer 1976 and November 1977 some 29000 spiders belonging to 168 species were caught in pitfalls on 42 peat and upland grassland sites covering an altitude range of 11–827 m in the north of England. The similarities between the spider faunas of each site have been assessed using a modified form of Sørensen's Index and the sites clustered according to their similarities. The sites divided into two major groups; (1) mineral soils where grasses are dominant or shallow peat soils where Juncus squarrosus is dominant and (2) peat soils where Eriophorum vaginatum and Calluna vulgaris are dominant. It is suggested that this major division is the result of the plant "architecture" rather than the species composition of the vegetation. Direct comparison between paired peat and grassland sites at the same altitudes shows no marked separation in the spider species composition on the two soil types but there are differences in abundance of individual species. The three peat site clusters form an altitudinal sequence which is not the result of the decline in numbers of non-linyphiid species with increase in altitude as the cluster formation is largely dependent on the linyphiid species which dominate the catches.
Twenty one common species have been used lo identify the communities. Many of these species are often found together and form associations. Typically, a number of associations contribute to each community. Only 19 species contribute more than 4% to the numbers of individuals caught in any one community.
Diversity declines with increase in altitude, as does the number of species caught. The decline in the number of species caught is the effect of a decrease in non-linyphiid species with increasing altitude. There is no significant decline in the numbers of species of linyphiids with increase in altitude.     

Herbivory in global climate change research: direct effects of rising temperature on insect herbivores

This review examines the direct effects of climate change on insect herbivores. Temperature is identified as the dominant abiotic factor directly affecting herbivorous insects. There is little evidence of any direct effects of CO₂ or UVB. Direct impacts of precipitation have been largely neglected in current research on climate change. Temperature directly affects development, survival, range and abundance. Species with a large geographical range will tend to be less affected. The main effect of temperature in temperate regions is to influence winter survival; at more northerly latitudes, higher temperatures extend the summer season, increasing the available thermal budget for growth and reproduction. Photoperiod is the dominant cue for the seasonal synchrony of temperate insects, but their thermal requirements may differ at different times of year. Interactions between photoperiod and temperature determine phenology; the two factors do not necessarily operate in tandem. Insect herbivores show a number of distinct life‐history strategies to exploit plants with different growth forms and strategies, which will be differentially affected by climate warming. There are still many challenges facing biologists in predicting and monitoring the impacts of climate change. Future research needs to consider insect herbivore phenotypic and genotypic flexibility, their responses to global change parameters operating in concert, and awareness that some patterns may only become apparent in the longer term.     

Proteomic identification of altered protein O-GlcNAcylation in a triple transgenic mouse model of Alzheimer's disease

PET scan analysis demonstrated the early reduction of cerebral glucose metabolism in Alzheimer disease (AD) patients that can make neurons vulnerable to damage via the alteration of the hexosamine biosynthetic pathway (HBP). Defective HBP leads to flawed protein O-GlcNAcylation coupled, by a mutual inverse relationship, with increased protein phosphorylation on Ser/Thr residues. Altered O-GlcNAcylation of Tau and APP have been reported in AD and is closely related with pathology onset and progression. In addition, type 2 diabetes patients show an altered O-GlcNAcylation/phosphorylation that might represent a link between metabolic defects and AD progression. Our study aimed to decipher the specific protein targets of altered O-GlcNAcylation in brain of 12-month-old 3×Tg-AD mice compared with age-matched non-Tg mice. Hence, we analysed the global O-GlcNAc levels, the levels and activity of OGT and OGA, the enzymes controlling its cycling and protein specific O-GlcNAc levels using a bi-dimensional electrophoresis (2DE) approach. Our data demonstrate the alteration of OGT and OGA activation coupled with the decrease of total O-GlcNAcylation levels. Data from proteomics analysis led to the identification of several proteins with reduced O-GlcNAcylation levels, which belong to key pathways involved in the progression of AD such as neuronal structure, protein degradation and glucose metabolism. In parallel, we analysed the O-GlcNAcylation/phosphorylation ratio of IRS1 and AKT, whose alterations may contribute to insulin resistance and reduced glucose uptake. Our findings may contribute to better understand the role of altered protein O-GlcNAcylation profile in AD, by possibly identifying novel mechanisms of disease progression related to glucose hypometabolism.     

The effects of the extracellular manganese concentration and variation of the interpulse delay time in the CPMG sequence on water exchange time across erythrocyte membranes

There has been broad disagreement in the literature regarding the dependence of water exchange times (Te) across erythrocyte membranes studied by the 1H-NMR Carr-Purcell-Meiboom-Gill (CPMG) pulse sequence on extracellular Mn2+ concentration. While some workers saw no change in Te with Mn2+, others reported a 35-50% decrease in Te with this extracellular paramagnetic relaxation agent. We present 1H-NMR evidence that a 30-50% change in Te can be produced by interdependence of the interpulse delay time of the CPMG pulse sequence and the external Mn2+ concentration. Such a large dependency is interpreted in terms of the diffusional effect as a major source. However, it is shown experimentally that if a large number of refocusing pi pulses are used, the observed transverse relaxation times are unaffected by Mn2+. Under these conditions excellent agreement of Te obtained in our study (13.0 +/- 0.64 ms (N = 36) at 21 degrees C) and that of 12.8 +/- 3.6 ms at 20-23 degrees C reported by the radiotracer method was found. Our findings suggest new and important implications for evaluating the previous reports of the 1H-NMR CPMG method concerning the [Mn2+] effect in the decrease of Te, and provide conditions where studies of water transport across erythrocyte membranes using this magnetic resonance method can be used with confidence.