Protein-based materials, toward a new level of structural control

Through billions of years of evolution nature has created and refined structural proteins for a wide variety of specific purposes. Amino acid sequences and their associated folding patterns combine to create elastic, rigid or tough materials. In many respects, nature’s intricately designed products provide challenging examples for materials scientists, but translation of natural structural concepts into bio-inspired materials requires a level of control of macromolecular architecture far higher than that afforded by conventional polymerization processes. An increasingly important approach to this problem has been to use biological systems for production of materials. Through protein engineering, artificial genes can be developed that encode protein-based materials with desired features. Structural elements found in nature, such as β-sheets and α-helices, can be combined with great flexibility, and can be outfitted with functional elements such as cell binding sites or enzymatic domains. The possibility of incorporating non-natural amino acids increases the versatility of protein engineering still further. It is expected that such methods will have large impact in the field of materials science, and especially in biomedical materials science, in the future

ORGANIZATIONAL ECONOMICS AND THE FOOD PROCESSING INDUSTRY

The food processing industry is dominated by large corporations. These firms play a critical role in forming the derived demand faced by agricultural producers, but little is understood about how these companies make strategic choices. Organizational economics provides a framework for exploring the firm\u27s decision process. However, several theories exist in this discipline, operating in fundamentally different ways. This paper examines the two prevalent organizational theories, Transaction Cost Economics and Agency Theory, through a study of the food processing industry. This sector is thoroughly analyzed in order to make predictions from each theory regarding the aspects of capital structure and firm expansion. With accounting data for a sample of food processing firms, these predictions are then tested empirically using an ICAPM model in a cross-section of expected stock returns. Our results indicate that Agency Theory is the relevant organizational model for food manufacturers, making it the appropriate tool for evaluating the actions of these firms in agricultural markets

Biomimetic spatial and temporal (4D) design and fabrication

We imagine the built environment of the future as a ‘bio-hybrid machine for living in’ that will sense and react to activities within the space in order to provide experiences and services that will elevate quality of life while coexisting seamlessly with humans and the natural environment. The study of Hierarchical design in biological materials has the potential to alter the way designers/ engineers/ crafts-men of the future engage with materials in order to realise such visions. We are ex-ploring this design approach using digital manufacturing technologies such as jac-quard weaving and 3D printing

Dynamics of Phase Behavior of a Polymer Blend Under Shear Flow

We study the dynamics of the phase behavior of a polymer blend in the presence of shear flow. By adopting a two fluid picture and using a generalization of the concept of material derivative, we construct kinetic equations that describe the phase behavior of polymer blends in the presence of external flow. A phenomenological form for the shear modulus for the blend is proposed. The study indicates that a nonlinear dependence of the shear modulus of the blend on the volume fraction of one of the species is crucial for a shift in the stability line to be induced by shear flow.Comment: 16 pages, late

Mutant methionyl-tRNA synthetase from bacteria enables site-selective N-terminal labeling of proteins expressed in mammalian cells

Newly synthesized cellular proteins can be tagged with a variety of metabolic labels that distinguish them from preexisting proteins and allow them to be identified and tracked. Many such labels are incorporated into proteins via the endogenous cellular machinery and can be used in numerous cell types and organisms. Though broad applicability has advantages, we aimed to develop a strategy to restrict protein labeling to specified mammalian cells that express a transgene. Here we report that heterologous expression of a mutant methionyl-tRNA synthetase from Escherichia coli permits incorporation of azidonorleucine (Anl) into proteins made in mammalian (HEK293) cells. Anl is incorporated site-selectively at N-terminal positions (in competition with initiator methionines) and is not found at internal sites. Site selectivity is enabled by the fact that the bacterial synthetase aminoacylates mammalian initiator tRNA, but not elongator tRNA. N-terminally labeled proteins can be selectively conjugated to a variety of useful probes; here we demonstrate use of this system in enrichment and visualization of proteins made during various stages of the cell cycle. N-terminal incorporation of Anl may also be used to engineer modified proteins for therapeutic and other applications

Educating for Discipleship in Consumer Culture: Promising Practices Rooted in the Pastoral Circle

Thesis advisor: Jane ReganAmerican society has been labeled a consumer culture. Consumer culture is not another term for materialism or a framework to explain one's relationship to money; it is an evolving ethos shaping our vision of ourselves, of our neighbor and the common good. The breadth and depth of commodification in the contemporary West informs the collective imagination in unprecedented ways. This dissertation brings together social science critique, educational tools and theological resources to create models for effective adult disciple building that are adequate for addressing challenges of a dominant culture's ideology and practices. Christian formation practices should heighten the Christian community's awareness of its role in dominant culture, both as inheritors of culture and as agents. This awareness requires transformation in many dimensions of one's being: a holistic discipleship. Jesus reminded his followers, "Where your treasure is there your heart will be also." One of the driving questions of this dissertation is: how can the Christian community wrestle ultimate concerns back from the consumer culture to the heart of God for the world? To address that question the discourse of the dissertation is interdisciplinary while maintaining an ultimate vision for an approach to educating for mature Christian discipleship. The dissertation is structured to include social analysis, a vision of alternatives to the dominant lifestyle promulgated by the consumer culture, and effective pathways toward achieving that vision. The first half of the dissertation analyzes the relationship of contemporary consumer culture and Christian experience. The sociological and historical descriptions of this phenomenon lead toward the question, what are the implications for religious identity and meaning-making in light of the consumerist context? Theological resources include the gospel of Luke, Dietrich Bonhoeffer and Gustavo Gutierrez for highlighting key dimensions of culturally responsive discipleship. There are also two brief cases presented of organizations who are attempting to live out promising approaches to Christian community in light of consumer culture patterns.The second half explores theories that can serve as a framework for Christian education practices. Transformative learning theory is introduced as a resource for cultivating awareness of underlying assumptions shaped by culture that are operative in adult decision making and worldview. Henriot and Holland's pastoral circle is described as a transformative learning tool. The dissertation moves toward a model of adapting the pastoral circle for educating congregations to think theologically about culture for the sake of personal transformation and social action.Thesis (PhD) — Boston College, 2011.Submitted to: Boston College. Graduate School of Arts and Sciences.Discipline: Religious Education and Pastoral Ministry

EPR Study of Spin Labeled Brush Polymers in Organic Solvents

Spin-labeled polylactide brush polymers were synthesized via ring-opening metathesis polymerization (ROMP), and nitroxide radicals were incorporated at three different locations of brush polymers: the end and the middle of the backbone, and the end of the side chains (periphery). Electron paramagnetic resonance (EPR) was used to quantitatively probe the macromolecular structure of brush polymers in dilute solutions. The peripheral spin-labels showed significantly higher mobility than the backbone labels, and in dimethylsulfoxide (DMSO), the backbone end labels were shown to be more mobile than the middle labels. Reduction of the nitroxide labels by a polymeric reductant revealed location-dependent reactivity of the nitroxide labels: peripheral nitroxides were much more reactive than the backbone nitroxides. In contrast, almost no difference was observed when a small molecule reductant was used. These results reveal that the dense side chains of brush polymers significantly reduce the interaction of the backbone region with external macromolecules, but allow free diffusion of small molecules

Cleavable Biotin Probes for Labeling of Biomolecules via Azide−Alkyne Cycloaddition

The azide−alkyne cycloaddition provides a powerful tool for bio-orthogonal labeling of proteins, nucleic acids, glycans, and lipids. In some labeling experiments, e.g., in proteomic studies involving affinity purification and mass spectrometry, it is convenient to use cleavable probes that allow release of labeled biomolecules under mild conditions. Five cleavable biotin probes are described for use in labeling of proteins and other biomolecules via azide−alkyne cycloaddition. Subsequent to conjugation with metabolically labeled protein, these probes are subject to cleavage with either 50 mM Na_2S_2O_4, 2% HOCH_2CH_2SH, 10% HCO_2H, 95% CF_3CO_2H, or irradiation at 365 nm. Most strikingly, a probe constructed around a dialkoxydiphenylsilane (DADPS) linker was found to be cleaved efficiently when treated with 10% HCO_2H for 0.5 h. A model green fluorescent protein was used to demonstrate that the DADPS probe undergoes highly selective conjugation and leaves a small (143 Da) mass tag on the labeled protein after cleavage. These features make the DADPS probe especially attractive for use in biomolecular labeling and proteomic studies

State-Selective Metabolic Labeling of Cellular Proteins

Transcriptional activity from a specified promoter can provide a useful marker for the physiological state of a cell. Here we introduce a method for selective tagging of proteins made in cells in which specified promoters are active. Tagged proteins can be modified with affinity reagents for enrichment or with fluorescent dyes for visualization. The method allows state-selective analysis of the proteome, whereby proteins synthesized in predetermined physiological states can be identified. The approach is demonstrated by proteome-wide labeling of bacterial proteins upon activation of the P_(BAD) promoter and the SoxRS regulon and provides a basis for analysis of more complex systems including spatially heterogeneous microbial cultures and biofilms

Core-Clickable PEG-Branch-Azide Bivalent-Bottle-Brush Polymers by ROMP: Grafting-Through and Clicking-To

The combination of highly efficient polymerizations with modular "click" coupling reactions has enabled the synthesis of a wide variety of novel nanoscopic tructures. Here we demonstrate the facile synthesis of a new class of clickable, branched nanostructures, polyethylene glycol (PEG)-branch-azide bivalent-brush polymers, facilitated by "graft-through" ring-opening metathesis polymerization of a branched norbornene-PEG-chloride macromonomer followed by halide-azide exchange. The resulting bivalent-brush polymers possess azide groups at the core near a polynorbornene backbone with PEG chains extended into solution; the structure resembles a unimolecular micelle. We demonstrate copper-catalyzed azide-alkre cycloaddition (CuAAC) "click-to" coupling of a photocleavable doxorubicin (DOX)-alkyne derivative to the azide core. The CuAAC coupling was quantitative across a wide range of nanoscopic sizes (similar to 6-similar to 50 nrn); UV photolysis of the resulting DOX-loaded materials yielded free DOX that was therapeutically effective against human cancer cells