Optimization of Optical and Mechanical Properties of Real Architecture for 3-Dimensional Tissue Equivalents: Towards Treatment of Limbal Epithelial Stem Cell Deficiency

Limbal epithelial stem cell (LESC) deficiency can cause blindness. Transplantation of cultured human limbal epithelial cells (hLE) on human amniotic membrane (HAM) can restore vision but clinical graft manufacture can be unreliable. We have developed a reliable and robust tissue equivalent (TE) alternative to HAM, Real Architecture for 3D Tissue (RAFT). Here, we aimed to optimize the optical and mechanical properties of RAFT TE for treatment of LESC deficiency in clinical application. The RAFT TE protocol is tunable; varying collagen concentration and volume produces differing RAFT TEs. These were compared with HAM samples taken from locations proximal and distal to the placental disc. Outcomes assessed were transparency, thickness, light transmission, tensile strength, ease of handling, degradation rates and suitability as substrate for hLE culture. Proximal HAM samples were thicker and stronger with poorer optical properties than distal HAM samples. RAFT TEs produced using higher amounts of collagen were thicker and stronger with poorer optical properties than those produced using lower amounts of collagen. The ‘optimal’ RAFT TE was thin, transparent but still handleable and was produced using 0.6 ml of 3 mg/ml collagen. Degradation rates of the ‘optimal’ RAFT TE and HAM were similar. hLE achieved confluency on ‘optimal’ RAFT TEs at comparable rates to HAM and cells expressed high levels of putative stem cell marker p63α. These findings support the use of RAFT TE for hLE transplantation towards treatment of LESC deficiency

OPENING KEYNOTE ADDRESS: Using Data to Understand Cultural Destruction

Brian I. Daniels, Ph.D, Penn Cultural Heritage Center, University of Pennsylvania Museum. Why is cultural heritage targeted in conflict? Under what circumstances? By whom? Today, due in part to the recent notorious instances of cultural destruction in the Middle East and North Africa, there is perhaps more attention among the broader scientific community than ever before about the phenomenon of cultural loss. At the same time, there are many significant data and analytical gaps. Little social science literature about cultural destruction exists and many critical questions—and avenues of research—are, as of yet, unstudied. A primary reason for this lack of attention is the absence of event-level data about cultural destruction that can test theories about this kind of violence. This presentation discusses some of the data needs that exist in order to develop a more robust understanding about cultural destruction and the problems and prospects for future analysis

Development of Improved Rhenium Coatings for Fluorine Engine Thrust Chambers

Coating trials were undertaken to evaluate the application of rhenium to carbon-carbon composite sheet by plasma spraying. Optimum spray parameters and coating thickness were identified for production of coatings free from continuous defects and with adequate adherence to the substrate. A tungsten underlayer was not beneficial and possibly detracted from coating integrity. Stress calculations indicated that the proposed operating cycle of the rocket engine would not cause spalling of the rhenium coating. Calculations indicated that permeation of gases through the coating would not be significant during the expected life of the thrust chamber. The feasibility of applying rhenium coatings by laser melting was also studied. Poor wetting of the composite surface by the liquid rhenium precluded production of uniform coatings. Borate/carborate fluxes did not improve wetting characteristics

Islands of conformational stability for Filopodia

Filopodia are long, thin protrusions formed when bundles of fibers grow outwardly from a cell surface while remaining closed in a membrane tube. We study the subtle issue of the mechanical stability of such filopodia and how this depends on the deformation of the membrane that arises when the fiber bundle adopts a helical configuration. We calculate the ground state conformation of such filopodia, taking into account the steric interaction between the membrane and the enclosed semiflexible fiber bundle. For typical filopodia we find that a minimum number of fibers is required for filopodium stability. Our calculation elucidates how experimentally observed filopodia can obviate the classical Euler buckling condition and remain stable up to several tens of . We briefly discuss how experimental observation of the results obtained in this work for the helical-like deformations of enclosing membrane tubes in filopodia could possibly be observed in the acrosomal reactions of the sea cucumber Thyone, and the horseshoe crab Limulus. Any realistic future theories for filopodium stability are likely to rely on an accurate treatment of such steric effects, as analysed in this work

Limbal Fibroblasts Maintain Normal Phenotype in 3D RAFT Tissue Equivalents Suggesting Potential for Safe Clinical Use in Treatment of Ocular Surface Failure.

Limbal epithelial stem cell deficiency can cause blindness, but transplantation of these cells on a carrier such as human amniotic membrane can restore vision. Unfortunately, clinical graft manufacture using amnion can be inconsistent. Therefore, we have developed an alternative substrate, Real Architecture for 3D Tissue (RAFT), which supports human limbal epithelial cells (hLE) expansion. Epithelial organization is improved when human limbal fibroblasts (hLF) are incorporated into RAFT tissue equivalent (TE). However, hLF have the potential to transdifferentiate into a pro-scarring cell type, which would be incompatible with therapeutic transplantation. The aim of this work was to assess the scarring phenotype of hLF in RAFT TEs in hLE+ and hLE- RAFT TEs and in nonairlifted and airlifted RAFT TEs. Diseased fibroblasts (dFib) isolated from the fibrotic conjunctivae of ocular mucous membrane pemphigoid (Oc-MMP) patients were used as a pro-scarring positive control against which hLF were compared using surrogate scarring parameters: matrix metalloproteinase (MMP) activity, de novo collagen synthesis, α-smooth muscle actin (α-SMA) expression, and transforming growth factor-β (TGF-β) secretion. Normal hLF and dFib maintained different phenotypes in RAFT TE. MMP-2 and -9 activity, de novo collagen synthesis, and α-SMA expression were all increased in dFib cf. normal hLF RAFT TEs, although TGF-β1 secretion did not differ between normal hLF and dFib RAFT TEs. Normal hLF do not progress toward a scarring-like phenotype during culture in RAFT TEs and, therefore, may be safe to include in therapeutic RAFT TE, where they can support hLE, although in vivo work is required to confirm this. dFib RAFT TEs (used in this study as a positive control) may be useful toward the development of an ex vivo disease model of Oc-MMP

If You Build It, They Will Come: What Students Say About Experiential Learning

Our purpose here is to explore one of the “natural experiments” cited by the Task Force: the Experiential Advantage (EA) program at the University of Denver’s Sturm College of Law (Denver Law). EA was developed as a part of a greater general focus on experiential learning and is built upon the three “Carnegie Apprenticeships” – “the intellectual or cognitive,” “the forms of expert practice,” and “identity and purpose.” It was implemented at Denver Law starting with students entering in August 2013. To explore this natural experiment, we took a particular route and did so for what we see as good reason. It is often the case with such curricular experiments that the views of students are missing. But of course, it makes little sense to neglect them because such changes are supposedly made for the students’ benefit. So, it seems more than appropriate to ask them – from their perspective – if a change worked, or improved matters, as it was designed to do. This article is a first report on the findings of an extensive case study — a three-year, survey-based, study of Denver Law students concerning the EA Program “natural experiment.” The findings should be of considerable interest to the legal community, given that there is general support for experiential learning across most law schools, but a study of this kind — one exploring student views on curricular innovation — has never been conducted before. This Article is divided into four sections. The first provides a general context for Denver Law’s efforts. The second outlines the study itself, and is followed by section three, which analyzes the reasons first-year students chose Denver Law and their interest in EA and experiential learning. The fourth section changes focus and turns to Denver Law students nearing the end of their legal education – 3 and 4Ls. These students, nearing the end of their legal education, were asked a series of “look back” questions asking them about their law school experience, including the EA program and experiential learning

Health Disparities Among Racial and Ethnic Minority Firefighters

ABSTRACT Background: Racial/ethnic minorities are substantially underrepresented in the fire service and this situation is unique when compared to similarly mentally and physically demanding and hazardous occupations such as the military and law enforcement. There is little systematic research to provide greater clarity about this significant disparity. The purpose of this study is to examine physical and behavioral health issues of racial/ethnic minority firefighters when compared to their white, non-Hispanic counterparts and potentially identify areas for future research that might assist in improving their substantial underrepresentation. Materials and Methods: We report the results of a secondary analysis of data combining the baseline evaluations of two different firefighter health studies, the Firefighter Injury and Risk Evaluation (FIRE) and Fuel 2 Fight (F2F) studies. Male career firefighters (N=1,404) were from 31 fire departments across the US and its territories. White, non-Hispanic firefighters comprised 72.5% of the sample (n=1,018) and 27.5% classified themselves as a racial/ethnic minority. Firefighters who agreed to participate comprised 94% (F2F) and 97% (FIRE) of those available and all underwent assessments including body composition, fitness, and general/behavioral health, and job satisfaction. Results: We examined differences in health and job status between minority and non-minority firefighters and between firefighters in minority- (MDCs) and white-dominated communities (WDCs). After adjusting for potential confounds, there were significant main effects for the individual minority status vs. non-minority status on both BMI and BF%, indicating that minority firefighters had significantly higher average BMI (28.8±0.3kg/m2) and BF% (24.7± 0.7%) when compared to their white, non-Hispanic colleagues (27.7±0.2kg/m2and 23.1±0.6% for BMI and BF%, respectively). Minority firefighters also were 59% more likely to be obese (adjusted [A]OR=1.59; 95% CI=1.16-2.18). Firefighters serving in MDCs reported significantly more poor health days (Mean±SE; 3.2±0.2 days) than firefighters serving in WDCs (2.8±0.2 days; p=0.038). In addition, minority firefighters reported significantly more poor health days (3.6±0.4 days) than their non-minority colleagues (2.8±0.2 days; p=0.003), while the interaction indicates that minority firefighters in MDCs reported more poor health days than the other groups (p Conclusions: Individual and community minority status (i.e., ethnic density effect) were both significantly associated with a number of important health status indicators, with racial/ethnic minority firefighters demonstrating greater risk for unfavorable body composition and more poor physical health days. In addition, minority firefighters in WDCs reported the highest prevalence of lifetime diagnosis of depression by a physician, while minority firefighters in MDCs had the lowest. Many of these health status indicators have recently been studied within the context of experiences with discrimination, demonstrating that racial discrimination is associated with greater risk for obesity, depression, and poor physical and mental health and could be contributing to health disparities and potentially negatively impacting racial/ethnic minority firefighter health, safety, and retention