The adventures of Rose and Jody, Chapter 17: Jody does her thesis

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Anionic lipid vesicles have differential effects on the aggregation of early onset-associated α-synuclein missense mutants

α-synuclein (αS) is the key component of synucleinopathies such as Parkinson’s disease (PD), dementia with Lewy bodies, and multiple system atrophy. αS was first linked to PD through the identification of point mutations in the SNCA gene, causing single amino acid substitutions within αS and familial autosomal dominant forms of PD that profoundly accelerated disease onset by up to several decades. At least eight single-point mutations linked to familial PD (A30G/P, E46K, H50Q, G51D, and A53T/E/V) are located in proximity of the region preceding the non-β amyloid component (preNAC) region, strongly implicating its pathogenic role in αS-mediated cytotoxicity. Furthermore, lipids are known to be important for native αS function, where they play a key role in the regulation of synaptic vesicle docking to presynaptic membranes and dopamine transmission. However, the role of lipids in the function of mutant αS is unclear. Here, we studied αS aggregation properties of WT αS and five of the most predominant single-point missense mutants associated with early onset PD in the presence of anionic 1,2-dimyristoyl-sn-glycero-3-phospho-l-serine lipid vesicles. Our results highlight significant differences between aggregation rates, the number of aggregates produced, and overall fibril morphologies of WT αS and the A30P, E46K, H50Q, G51D, and A53T missense mutants in the presence of lipid vesicles. These findings have important implications regarding the interplay between the lipids required for αS function and the individual point mutations known to accelerate PD and related diseases

The Library Derived 4554W Peptide Inhibits Primary Nucleation of α-Synuclein

Aggregation of alpha-Synuclein (aS) is widely regarded as a key factor in neuronal cell death, leading to a wide range of synucleinopathies that includes Parkinson’s Disease. Development of therapeutics has therefore focused on inhibiting aggregation of aS into toxic forms. One such inhibitor, based on the preNAC region aS45-54 (4554W), was identified using an intracellular peptide library screen, and subsequently shown to both inhibit formation of aS aggregates while simultaneously lowering toxicity. Subsequent efforts have sought to determine the mode of 4554W action. In particular, and consistent with the fact that both target and peptide are co-produced during library screening, we find that the peptide inhibits primary nucleation of aS, but does not modulate downstream secondary nucleation or elongation events. These findings hold significant promise towards mechanistic understanding and development of molecules that can module the first steps in aS aggregation towards novel treatments for Parkinson’s disease and related synucleinopathies

Library-derived peptide aggregation modulators of Parkinson's disease early-onset alpha-synuclein variants.

[Image: see text] Parkinson’s Disease (PD) is characterized by the accumulation of Lewy bodies in dopaminergic neurons. The main protein component of Lewy bodies, α-synuclein (αS), is also firmly linked to PD through the identification of a number of single point mutations that are autosomal dominant for early-onset disease. Consequently, the misfolding and subsequent aggregation of αS is thought to be a key stage in the development and progression of PD. Therefore, modulating the aggregation pathway of αS is an attractive therapeutic target. Owing to the fact that all but one of the familial mutations is located in the preNAC 45–54 region of αS, we previously designed a semi-rational library using this sequence as a design scaffold. The 45–54 peptide library was screened using a protein-fragment complementation assay approach, leading to the identification of the 4554W peptide. The peptide was subsequently found to be effective in inhibiting primary nucleation of αS, the earliest stage of the aggregation pathway. Here, we build upon this previous work by screening the same 45–54 library against five of the known αS single-point mutants that are associated with early-onset PD (A30P, E46K, H50Q, G51D, and A53T). These point mutations lead to a rapid acceleration of PD pathology by altering either the rate or type of aggregates formed. All ultimately lead to earlier disease onset and were therefore used to enforce increased assay stringency during the library screening process. The ultimate aim was to identify a peptide that is effective against not only the familial αS variant from which it has been selected but that is also effective against WT αS. Screening resulted in five peptides that shared common residues at some positions, while deviating at others. All reduced aggregation of the respective target, with several also identified to be effective at reducing aggregation when incubated with other variants. In addition, our results demonstrate that a previously optimized peptide, 4554W(N6A), is highly effective against not only WT αS but also several of the single-point mutant forms and hence is a suitable baseline for further work toward a PD therapeutic

An N-terminal alpha-Synuclein fragment binds lipid vesicles to modulate lipid induced aggregation

Misfolding and aggregation of alpha-synuclein (αS) into toxic conformations is involved in numerous neurodegenerative diseases. In Parkinson's disease (PD), this occurs within dopaminergic neurons, causing cell death and disease symptoms. During αS aggregation, many protein-protein interactions (PPIs) form over broad and flat protein surfaces, limiting potential for small-molecule intervention. Peptides, however, harbor great therapeutic promise since they can selectively engage with and modulate the large surface areas involved yet are small enough to function as druggable agents if suitably structured. Here, we explore the first 25 residues of αS (αS 1–25) as a template for peptide-based αS aggregation antagonists. We report that αS 1–25 inhibits lipid-induced αS aggregation in a dose-dependent manner. αS 1–25 functions by binding to lipids to prevent αS binding, with both αS and peptide requiring lipid for inhibition to occur. These findings present a potential mechanistic route for the treatment or prevention of PD.</p

A downsized and optimised intracellular library-derived peptide prevents alpha-synuclein primary nucleation and toxicity without impacting upon lipid binding

Misfolding and aggregation of alpha-synuclein (αS) within dopaminergic neurons is a key factor in the development and progression of a group of age-related neurodegenerative diseases, termed synucleinopathies, that include Parkinson's disease (PD). We previously derived a peptide inhibitor from a 209,952-member intracellular library screen by employing the preNAC region (45–54) as a design template. At least six single-point mutations firmly linked to early-onset Parkinson's disease (E46K, H50Q, G51D, A53T/E/V) are located within this region, strongly implicating a pathogenic role within αS that leads to increased cytotoxicity. A library-derived ten residue peptide, 4554W, was consequently shown to block αS aggregation at the point of primary nucleation via lipid induction, inhibiting its conversion into downstream cytotoxic species. Here we couple truncation with a full alanine scan analysis, to establish the effect upon the αS aggregation pathway relative to 4554W. This revealed the precise residues responsible for eliciting inhibitory interaction and function, as well as those potentially amenable to modification or functionalisation. We find that modification N6A combined with N-terminal truncation results in a peptide of significantly increased efficacy. Importantly, our data demonstrate that the peptide does not directly disrupt αS lipid-binding, a desirable trait since antagonists of αS aggregation and toxicity should not impede association with small synaptic neurotransmitter vesicles, and thus not disrupt dopaminergic vesicle fusion and recycling. This work paves the way toward the major aim of deriving a highly potent peptide antagonist of αS pathogenicity without impacting on native αS function.</p

Single-Player and Two-Player Buttons & Scissors Games

We study the computational complexity of the Buttons \& Scissors game and obtain sharp thresholds with respect to several parameters. Specifically we show that the game is NP-complete for $C = 2$ colors but polytime solvable for $C = 1$. Similarly the game is NP-complete if every color is used by at most $F = 4$ buttons but polytime solvable for $F \leq 3$. We also consider restrictions on the board size, cut directions, and cut sizes. Finally, we introduce several natural two-player versions of the game and show that they are PSPACE-complete.Comment: 21 pages, 15 figures. Presented at JCDCG2 2015, Kyoto University, Kyoto, Japan, September 14 - 16, 201

Paradigms for Creating Activities that Integrate Mathematics and Science Topics

Research has shown that undergraduate students benefit from seeing examples of mathematics applied to real-world situations. This article describes three different paradigms for how math and discipline partner faculty worked together to create mathematical activities that illustrate applications of the topics being studied in precalculus and calculus. All three examples are discussed within the framework of PDSA cycles to describe the process by which the teams collaborated to plan, enact, study, and refine their lessons. Findings discuss both the difficulties of creating integrated activities (differences in terms and definitions between math and science faculty, different foregrounding of math versus science among faculty), and the value of the resultant lessons, such as increased level of student engagement, higher cognitive demand, and the role that relevant applications can play in piquing student interest in STEM

The Impact of a Youth Development Program on Secondary Students’ Career Aspirations

This study’s purpose determined the extent to which adolescents’ participation in a youth development program may be linked to the participants’ post-secondary education and career aspirations. One hundred and seven adolescents, ages 14-19 in grades 8-12 completed Holland’s Vocational Interest Survey and the 4-H Career Decision Survey. Ordinal regression analysis indicated participation in 4-H had a positive impact on career decisions for students who participated in 4-H for two years (p< .038) and six years (p< .001). Significant differences were apparent with 80% of the racial/ethnic groups surveyed concerning a college fair’s impact on career choice and college major determination