Diffusion Magnetic Resonance Imaging-Based Biomarkers for Neurodegenerative Diseases

There has been an increasing prevalence of neurodegenerative diseases with the rapid increase in aging societies worldwide. Biomarkers that can be used to detect pathological changes before the development of severe neuronal loss and consequently facilitate early intervention with disease-modifying therapeutic modalities are therefore urgently needed. Diffusion magnetic resonance imaging (MRI) is a promising tool that can be used to infer microstructural characteristics of the brain, such as microstructural integrity and complexity, as well as axonal density, order, and myelination, through the utilization of water molecules that are diffused within the tissue, with displacement at the micron scale. Diffusion tensor imaging is the most commonly used diffusion MRI technique to assess the pathophysiology of neurodegenerative diseases. However, diffusion tensor imaging has several limitations, and new technologies, including neurite orientation dispersion and density imaging, diffusion kurtosis imaging, and free-water imaging, have been recently developed as approaches to overcome these constraints. This review provides an overview of these technologies and their potential as biomarkers for the early diagnosis and disease progression of major neurodegenerative diseases.     

Exploratory factor analysis and validity of the virtual reality symptom questionnaire and computer use survey

Abstract

The widespread use of virtual reality head-mounted-displays (HMDs) calls for a re-examination of the impact of prolonged exposure to fixed visual displays at close ocular proximity. The purpose of this study is to validate the Virtual Reality Symptoms Questionnaire (VRSQ), created to understand symptoms of prolonged HMDs use, and Computer Use Survey (CUS), created to assess general physical and visual discomfort symptoms. Participants (N?=?100) recorded their general discomfort symptoms using the CUS, performed an interactive task using a HMD for thirty minutes, and then answered the CUS again along with the VRSQ. VRSQ, analysed using an exploratory factor analysis, indicated a clear two-factor solution, and demonstrated very good internal consistency (α?=?0.873). The CUS, also analysed using an exploratory factor analysis, indicated a four-factor solution, and demonstrated good internal consistency (α?=?0.838).

Practitioner Summary: A quantitative-experimental study was conducted to explore the factor structure and validate both the Virtual Reality Symptoms Questionnaire (VRSQ), and the Computer Use Survey (CUS). Findings indicate the VRSQ and CUS are precise and accurate survey instruments for evaluating discomfort after VR-HMD use and the latter for computer use.

Abbreviations: VRSQ: virtual reality symptom questionnaire; CUS: computer use survey; OLED: organic light-emitting diode; MSQ: pensacola motion symptom questionnaire; SSQ: simulator sickness questionnaire; 3?D: three-dimensional computer generated space; VR: virtual reality; VR-HMD: virtual reality head-mounted-display; HMDs: head-mounted-displays; EFA: exploratory factor analysis     

Molecular Sensing Using Hyperpolarized Xenon NMR Spectroscopy

Molecular imaging is the determination of the spatial location and concentration of specific molecules in a sample of interest. Sophisticated modern magnetic resonance imaging machines can collect NMR spectra from small-volume elements within a sample, enabling local chemical analysis. However, abundant water and fat signals limit detection of metabolites to near mM concentrations. Alternatively, targeted relaxation contrast agents enhance the relaxation of the strong water signal where they bind. A comparison of images with and without a contrast agent shows the target distribution, but high µM concentrations are needed. We have developed an approach that exploits the strong signals of hyperpolarized ¹²⁹Xe (an inert reporter introduced for imaging). The imaging contrast agents are composed of a biological recognition motif to localize the agent (antibodies or aptamers) and covalently tethered cryptophane cages. Xenon binds to the cryptophane and though chemical exchange saturation transfer creates contrast in a xenon image. Imaging agents can deliver many cages per target, giving detection limits in the pM concentration range. The evolution and principles of this approach are described herein.     

Photo-mediated copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) “click” reactions for forming polymer networks as shape memory materials

The formation of polymer networks polymerized with the Copper (I) – catalyzed azide – alkyne cycloaddition (CuAAC) click reaction is described along with their accompanying utilization as shape memory polymers. Due to the click nature of the reaction and the synthetic accessibility of azide and alkyne functional-monomers, the polymer architecture was readily controlled through monomer design to manipulate crosslink density, ability for further functionalization, and the glass transition temperature (55–114 °C). Free strain recovery is used to quantify the shape memory properties of a model CuAAC network resulting in excellent shape fixity and recovery of 99%. The step growth nature of this polymerization results in homogenous network formation with narrow glass transitions ranges having half widths of the transition close to 15 °C for these materials resulting in shape recovery sharpness of 3.9%/°C in a model system comparable to similarly crosslinked chain growth polymers. Utilization of the CuAAC reaction to form shape memory materials opens a range of possibilities and behaviors that are not readily achieved in other shape memory materials such as (meth) acrylates, thiol-ene, thiol-Michael, and poly(caprolactone) based shape memory materials.     

Effects of 3‐D microwell culture on initial fate specification in human embryonic stem cells

Several studies have demonstrated that three‐dimensional (3‐D) culture systems influence human embryonic stem cell (hESC) phenotypes and fate choices. However, the effect that these microenvironmental changes have on signaling pathways governing hESC behaviors is not well understood. Here, a 3‐D microwell array was used to investigate differences in activation of developmental pathways between 2‐D and 3‐D cultures of both undifferentiated hESCs and hESCs undergoing initial differentiation in embryoid bodies (EBs). An increased induction into mesoderm and endoderm and differences in expression of genes from multiple signaling pathways that regulate development, including Wnt/β‐catenin, TGF‐β superfamily, Notch, and FGF during EB‐mediated differentiation were observed in 3‐D microwells as compared with the 2‐D substrates. In undifferentiated hESCs, differences in epithelial‐mesenchymal transition phenotypes and the TGFβ/BMP pathway between cultures in 3‐D and 2‐D were also observed. These results illustrate that 3‐D culture influences multiple pathways that may regulate the differentiation trajectories of hESCs. © 2014 American Institute of Chemical Engineers AIChE J, 60: 1225–1235, 2014     

Photobleaching Reveals Heterogeneous Stoichiometry for Equinatoxin II Oligomers

Equinatoxin II (EqtII), a sea anemone cytolysin, is known to oligomerize to form pores that spontaneously insert into membranes. Crystallographic and cryo‐EM studies of structurally similar cytolysins offer contradictory evidence for pore stoichiometry. Here we used single‐molecule photobleaching of fluorescently labeled EqtII to determine the stoichiometry of EqtII oligomers in supported lipid bilayers. A frequency analysis of photobleaching steps revealed a log‐normal distribution of stoichiometries with a mean of 3.4±2.3 standard deviations. Comparison of our experimental data with simulations of fixed stoichiometries supports our observation of a heterogeneous distribution of EqtII oligomerization. These data are consistent with a model of EqtII stoichiometry where pores are on average tetrameric, but with large variation in the number of subunits in individual pores.     

Catalytic Molecular Logic Devices by DNAzyme Displacement

Chemical reactions catalyzed by DNAzymes offer a route to programmable modification of biomolecules for therapeutic purposes. To this end, we have developed a new type of catalytic DNA‐based logic gates in which DNAzyme catalysis is controlled via toehold‐mediated strand displacement reactions. We refer to these as DNAzyme displacement gates. The use of toeholds to guide input binding provides a favorable pathway for input recognition, and the innate catalytic activity of DNAzymes allows amplification of nanomolar input concentrations. We demonstrate detection of arbitrary input sequences by rational introduction of mismatched bases into inhibitor strands. Furthermore, we illustrate the applicability of DNAzyme displacement to compute logic functions involving multiple logic gates. This work will enable sophisticated logical control of a range of biochemical modifications, with applications in pathogen detection and autonomous theranostics.     

Identification and Characterization of a Welwitindolinone Alkaloid Biosynthetic Gene Cluster in the Stigonematalean Cyanobacterium Hapalosiphon welwitschii

The identification of a 36 kb welwitindolinone (wel) biosynthetic gene cluster in Hapalosiphon welwitschii UTEX B1830 is reported. Characterization of the enzymes responsible for assembling the early biosynthetic intermediates geranyl pyrophosphate and 3‐((Z)‐2′‐isocyanoethenyl)indole as well as a dedicated N‐methyltransferase in the maturation of N‐methylwelwitindolinone C isothiocyanate solidified the link between the wel pathway and welwitindolinone biosynthesis. Comparative analysis of the ambiguine and welwitindolinone biosynthetic pathways in two different organisms provided insights into the origins of diverse structures within hapalindole‐type molecules.