Raman Spectroscopy with 2D Perturbation Correlation Moving Windows for the Characterization of Heparin-Amyloid Interactions.

It has been shown extensively that glycosaminoglycan (GAG)-protein interactions can induce, accelerate, and impede the clearance of amyloid fibrils associated with systemic and localized amyloidosis. Obtaining molecular details of these interactions is fundamental to our understanding of amyloid disease. Consequently, there is a need for analytical approaches that can identify protein conformational transitions and simultaneously characterize heparin interactions. By combining Raman spectroscopy with two-dimensional (2D) perturbation correlation moving window (2DPCMW) analysis, we have successfully identified changes in protein secondary structure during pH- and heparin-induced fibril formation of apolipoprotein A-I (apoA-I) associated with atherosclerosis. Furthermore, from the 2DPCMW, we have identified peak shifts and intensity variations in Raman peaks arising from different heparan sulfate moieties, indicating that protein-heparin interactions vary at different heparin concentrations. Raman spectroscopy thus reveals new mechanistic insights into the role of GAGs during amyloid fibril formation.

Stage Salience and Situational Likelihood in the Formation of Situation Models during Sentence Comprehension.

Two experiments examined the relation between event structure, situational likelihood and eye fixation time while reading predicate modifiers in isolated sentences. Experiment 1 used activity predicates and preparatory process predicates (climbed a mountain), which make salient the process that leads to a culmination. Preparatory process predicates increased first pass time on durative modifiers (for several years) and decreased total time on frequency modifiers (e.g., every year). Situational likelihood was associated with fixation times on frame modifiers (last year) but not with fixation times on durative or frequency modifiers. Experiment 2 used activity predicates and result state predicates (halted a class), which make salient the result that follows from a culmination. Result state predicates had no effect on fixation times on durative modifiers and decreased total time on frequency modifiers. Situational likelihood was associated only with total time on durative modifiers. These results demonstrate that readers use the meanings of predicates and modifiers to form an initial model of a sentence and that the likelihood of the reported situation is related to reading time relatively late. The results are discussed in terms of type coercion theory and situation models in sentences and narratives.

Aspectual coercion in eye movements.

Comprehension includes interpreting sentences in terms of aspectual categories such as processes (Harry climbed) and culminations (Harry reached the top). Adding a verbal modifier such as for many years to a culmination coerces its interpretation from one to many culminations. Previous studies have found that coercion increases lexical decision and meaning judgment time, but not eye fixation time. This study recorded eye movements as participants read sentences in which a coercive adverb increased the interpretation of multiple events. Adverbs appeared at the end of a clause and line; the post-adverb region appeared at the beginning of the next line; follow-up questions occasionally asked about aspectual meaning; and clause type varied systematically. Coercive adverbs increased eye fixation time in the post-adverb region and in the adverb and post-adverb regions combined. Factors that influence the appearance of aspectual coercion may include world knowledge, follow-up questions, and the location and ambiguity of adverbs.

Circular Dichroism Spectroscopy Identifies the ß-Adrenoceptor Agonist Salbutamol As a Direct Inhibitor of Tau Filament Formation in Vitro.

Potential drug treatments for Alzheimer's disease (AD) may be found by identifying compounds that block the assembly of the microtubule-associated protein tau into neurofibrillar tangles associated with neuron destabilization and cell death. Here, a small library of structurally diverse compounds was screened in vitro for the ability to inhibit tau aggregation, using high-throughput synchrotron radiation circular dichroism as a novel tool to monitor the structural changes in the protein as it assembles into filaments. The catecholamine epinephrine was found to be the most effective tau aggregation inhibitor of all 88 screened compounds. Subsequently, we tested chemically similar phenolamine drugs from the ß-adrenergic receptor agonist class, using conventional circular dichroism spectroscopy, thioflavin T fluorescence, and transmission electron microscopy. Two compounds, salbutamol and dobutamine, used widely in the treatment of respiratory and cardiovascular disease, impede the aggregation of tau in vitro. Dobutamine reduces both the rate and yield of tau filament formation over 24 h; however, it has little effect on the structural transition of tau into ß-sheet structures over 24 h. Salbutamol also reduces the yield and rate of filament formation and additionally inhibits tau's structural change into ß-sheet-rich aggregates. Salbutamol has a good safety profile and a half-life that facilitates permeation through the blood-brain barrier and could represent an expediated approach to developing AD therapeutics. These results provide the motivation for the in vivo evaluation of pre-existing ß-adrenergic receptor agonists as a potential therapy for AD through the reduction of tau deposition.