Achieving Acetylcholine Receptor Clustering in Tissue-Engineered Skeletal Muscle Constructs In vitro through a Materials-Directed Agrin Delivery Approach

Volumetric muscle loss (VML) can result from trauma, infection, congenital anomalies, or surgery, and produce permanent functional and cosmetic deficits. There are no effective treatment options for VML injuries, and recent advances toward development of muscle constructs lack the ability to achieve innervation necessary for long-term function. We sought to develop a proof-of-concept biomaterial construct that could achieve acetylcholine receptor (AChR) clustering on muscle-derived cells (MDCs) in vitro. The approach consisted of the presentation of neural (Z+) agrin from the surface of microspheres embedded with a fibrin hydrogel to muscle cells (C2C12 cell line or primary rat MDCs). AChR clustering was spatially restricted to areas of cell (C2C12)-microsphere contact when the microspheres were delivered in suspension or when they were incorporated into a thin (2D) fibrin hydrogel. AChR clusters were observed from 16 to 72 h after treatment when Z+ agrin was adsorbed to the microspheres, and for greater than 120 h when agrin was covalently coupled to the microspheres. Little to no AChR clustering was observed when agrin-coated microspheres were delivered from specially designed 3D fibrin constructs. However, cyclic stretch in combination with agrin-presenting microspheres led to dramatic enhancement of AChR clustering in cells cultured on these 3D fibrin constructs, suggesting a synergistic effect between mechanical strain and agrin stimulation of AChR clustering in vitro. These studies highlight a strategy for maintaining a physiological phenotype characterized by motor endplates of muscle cells used in tissue engineering strategies for muscle regeneration. As such, these observations may provide an important first step toward improving function of tissue-engineered constructs for treatment of VML injuries

Computerized adaptive testing of symptom severity : a registry-based study of 924 patients with trapeziometacarpal arthritis

We aimed to develop a computerized adaptive testing (CAT) version of the 11 item Patient Evaluation Measure (PEM), using an item response theory model. This model transformed the ordinal scores into ratio-interval scores. We obtained PEM responses from 924 patients with trapeziometacarpal osteoarthritis to build a CAT model and tested its performance on a simulated cohort of 1000 PEM response sets. The CAT achieved high precision (median standard error or measurement 0.26) and reduced the number of questions needed for accurate scoring from 11 to median two. The CAT scores and item-response-theory-based 15-item PEM scores were similar, and a Bland–Altman analysis demonstrated a mean score difference of 0.2 between the CAT and the full-length PEM scores on a scale from 0 to 100. We conclude that the CAT substantially reduced the burden of the PEM while also harnessing the validity of item response theory scoring

Discovery and Optimisation of a Compound Series active against Trypanosoma cruzi, the causative agent of Chagas’ Disease

Chagas disease is caused by the protozoan parasite; Trypanosoma cruzi; . It is endemic in South and Central America and recently has been found in other parts of the world, due to migration of chronically infected patients. The current treatment for Chagas disease is not satisfactory, and there is a need for new treatments. In this work, we describe the optimization of a hit compound resulting from the phenotypic screen of a library of compounds against; T. cruzi; . The compound series was optimized to the level where it had satisfactory pharmacokinetics to allow an efficacy study in a mouse model of Chagas disease. We were able to demonstrate efficacy in this model, although further work is required to improve the potency and selectivity of this series

The Ekpyrotic Universe: Colliding Branes and the Origin of the Hot Big Bang

We propose a cosmological scenario in which the hot big bang universe is produced by the collision of a brane in the bulk space with a bounding orbifold plane, beginning from an otherwise cold, vacuous, static universe. The model addresses the cosmological horizon, flatness and monopole problems and generates a nearly scale-invariant spectrum of density perturbations without invoking superluminal expansion (inflation). The scenario relies, instead, on physical phenomena that arise naturally in theories based on extra dimensions and branes. As an example, we present our scenario predominantly within the context of heterotic M-theory. A prediction that distinguishes this scenario from standard inflationary cosmology is a strongly blue gravitational wave spectrum, which has consequences for microwave background polarization experiments and gravitational wave detectors.Comment: 67 pages, 4 figures. v2,v3: minor corrections, references adde

Ferritins: furnishing proteins with iron

Ferritins are a superfamily of iron oxidation, storage and mineralization proteins found throughout the animal, plant, and microbial kingdoms. The majority of ferritins consist of 24 subunits that individually fold into 4-α-helix bundles and assemble in a highly symmetric manner to form an approximately spherical protein coat around a central cavity into which an iron-containing mineral can be formed. Channels through the coat at inter-subunit contact points facilitate passage of iron ions to and from the central cavity, and intrasubunit catalytic sites, called ferroxidase centers, drive Fe2+ oxidation and O2 reduction. Though the different members of the superfamily share a common structure, there is often little amino acid sequence identity between them. Even where there is a high degree of sequence identity between two ferritins there can be major differences in how the proteins handle iron. In this review we describe some of the important structural features of ferritins and their mineralized iron cores and examine in detail how three selected ferritins oxidise Fe2+ in order to explore the mechanistic variations that exist amongst ferritins. We suggest that the mechanistic differences reflect differing evolutionary pressures on amino acid sequences, and that these differing pressures are a consequence of different primary functions for different ferritins

Comparison of Direct Acoustic Impingement of Avionics Equipment to Vibration Base Shake Response

No abstract availabl

Tyr25, Tyr58 and Trp133 of Escherichia coli bacterioferritin transfer electrons between iron in the central cavity and the ferroxidase centre

Ferritins are 24meric proteins that overcome problems of toxicity, insolubility and poor bioavailability of iron in all types of cells by storing it in the form of a ferric mineral within their central cavities. In the bacterioferritin (BFR) from Escherichia coli iron mineralization kinetics have been shown to be dependent on an intra-subunit catalytic diiron cofactor site (the ferroxidase centre), three closely located aromatic residues and an inner surface iron site. One of the aromatic residues, Tyr25, is the site of formation of a transient radical, but the roles of the other two residues, Tyr58 and Trp133, are unknown. Here we show that these residues are important for the rates of formation and decay of the Tyr25 radical and decay of a secondary radical observed during Tyr25 radical decay. The data support a mechanism in which these aromatic residues function in electron transfer from the inner surface site to the ferroxidase centre