A Biomimetic Approach for the Creation of Two-Dimensional Microscale Surface Patterns: Creation of Isolated Immunological Synapses

Current efforts in surface functionalization have not produced a robust technique capable of creating specific two-dimensional microscale geometrical arrays composed of multiple proteins. Such a capability is desirable for engineering substrates in sensing and cell patterning applications where at least two different protein functionalities in a specific configuration are required. Here we introduce a new approach for the creation of arrays of microscale geometries. We demonstrate our approach with a biomimetic structure inspired by the immunological synapse, a cell-cell interfacial structure characterized by two concentric rings of proteins: an outer adhesion protein structure and an inner recognition ligand core. The power of the technique lies in its ability to pattern any protein in any defined geometry as well as to create arrays in parallel

Increased TCR Avidity after T Cell Activation A Mechanism for Sensing Low-Density Antigen

AbstractWhile activated T cells are known to have enhanced biological responses to antigen stimulation, the biophysical basis of this increased sensitivity remains unknown. Here, we show that, on activated T cells, the TCR avidity for peptide-MHC complexes is 20- to 50-fold higher than the TCR avidity of naive T cells. This increased avidity for peptide-MHC depends on TCR reorganization and is sensitive to the cholesterol content of the T cell membrane. Analysis of the binding data indicates the enhanced avidity is due to increases in cross-linking of TCR on activated T cells. Activation-induced membrane (AIM) changes in TCR avidity represent a previously unrecognized means of increasing the sensitivity of activated T cells to small amounts of antigen in the periphery

Dynamic scheduling model for the construction industry

Purpose:Basic project control through traditional methods is not sufficient to manage the majority of realtime events in most construction projects. This paper proposes a Dynamic Scheduling (DS) model that utilizes multi-objective optimization of cost, time, resources and cashflow, throughout project construction.Design/methodology/approach:Upon reviewing the topic of Dynamic Scheduling, a worldwide Internet survey with 364 respondents was conducted to define end-user requirements. The model was formulated and solution algorithms discussed. Verification was reported using predefined problem sets and a real-life case. Validation was performed via feedback from industry experts.Findings:The need for multi-objective dynamic software optimization of construction schedules and the ability to choose among a set of optimal alternatives were highlighted. Model verification through well-known test cases and a real-life project case study showed that the model successfully achieved the required dynamic functionality whether under the small solved example or under the complex case study. The model was validated for practicality, optimization of various DS schedule quality gates, ease of use, and software integration with contemporary project management practices.Practical/Social implications:Optimized real-time scheduling can provide better resources management including labour utilization and cost efficiency. Furthermore, DS contributes to optimum materials procurement, thus minimizing waste.Originality/value:The paper illustrates the importance of DS in construction, identifies the user needs, and overviews the development, verification and validation of a model that supports the generation of high quality schedules beneficial to large scale projects.</div

Influence of Shielding Gas Composition on the Properties of Flux-Cored Arc Welds of Plain Carbon Steel

In this study, the influence of variation in the shielding gas composition on the weld properties of steel ST 37-3 was investigated. Six different shielding gas compositions in addition to pure argon (Ar) and pure CO2 were studied in this work using flux cored arc welding (FCAW) process. For bead-on-plate (B.O.P) specimens pure Ar shielding gas has excellent arc stability; however, with increase of the CO2 percent in the shielding gas compositions, the arc stability becomes noisy (unstable arc) especially for pure CO2. 75% Ar – 25% CO2 shielding gas composition has the optimum deposition rate among other shielding gases for B.O.P. Furthermore, for complete real welded joints, the absorbed energy in the Charpy impact toughness test of weld metal (W.M) decreases with increase of the CO2 percent in the shielding gas composition. Additionally, the hardness of W.M decreases with the increase of the CO2 percent in the shielding gas composition

Enhanced Antigen-Specific Antitumor Immunity with Altered Peptide Ligands that Stabilize the MHC-Peptide-TCR Complex

AbstractT cell responsiveness to an epitope is affected both by its affinity for the presenting MHC molecule and the affinity of the MHC-peptide complex for TCR. One limitation of cancer immunotherapy is that natural tumor antigens elicit relatively weak T cell responses, in part because high-affinity T cells are rendered tolerant to these antigens. We report here that amino acid substitutions in a natural MHC class I–restricted tumor antigen that increase the stability of the MHC-peptide-TCR complex are significantly more potent as tumor vaccines. The improved immunity results from enhanced in vivo expansion of T cells specific for the natural tumor epitope. These results indicate peptides that stabilize the MHC-peptide-TCR complex may provide superior antitumor immunity through enhanced stimulation of specific T cells

Paracrine signalling events in embryonic stem cell renewal mediated by affinity targeted nanoparticles

AbstractStem cell growth and differentiation is controlled by intrinsic and extrinsic factors. The latter includes growth factors, which are conventionally supplied in vitro in media exchanged daily. Here, we illustrate the use of affinity targeted biodegradable nanoparticles to mediate paracrine stimulation as an alternative approach to sustain the growth and pluripotency of mouse embryonic stem cells. Leukaemia Inhibitory Factor (LIF) was encapsulated in biodegradable nanoparticles and targeted to the cell surface using an antibody to the oligosaccharide antigen SSEA-1. Sustained release of LIF from nanoparticles composed of a solid Poly(lactide-co-glycolic acid) polyester or a hydrogel-based liposomal system, we term Nanolipogel, replenished once after each cell passage, proved as effective as daily replenishment with soluble LIF for maintenance of pluripotency after 5 passages using 104-fold less LIF. Our study constitutes an alternative paradigm for stem cell culture, providing dynamic microenvironmental control of extrinsic bioactive factors benefiting stem cell manufacturing

Multiple Sclerosis: LIFNano-CD4 for Trojan Horse Delivery of the Neuro-Protective Biologic “LIF” Into the Brain: Preclinical Proof of Concept

Multiple sclerosis (MS) is a demyelinating autoimmune disease that attacks the brain, with year-on-year loss of brain volume, starting late teens and becoming manifest late twenties. There is no cure, and current therapies are immunosuppressive only. LIF is a vital stem cell growth factor active throughout life—and essential for health of the central nervous system (CNS), being tolerogenic, myelinogenic, and neuroprotective. Nano-formulation of LIF (LIFNano) using FDA-approved PLGA captures LIF's compound therapeutic properties, increasing potency 1,000-fold when targeted to CD4 (LIFNano-CD4). Moreover, circulating CD4+ lymphocytes are themselves regulated by LIF to express the Treg phenotype, known to release T cell-derived LIF upon engagement with cognate antigen, perpetuating antigen-specific self-tolerance. With the longer-term aim of treating inflammatory lesions of MS, we asked, does LIFNano-CD4 cross the blood–brain barrier (BBB)? We measure pK and pD using novel methodologies, demonstrate crossing of the BBB, show LIF-cargo-specific anti-inflammatory efficacy in the frontal cortex of the brain, and show safety of intravenous delivery of LIFNano-CD4 at doses known to provide efficacious concentrations of LIF cargo behind the BBB