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Immuno-oncology approaches that utilize T cell receptors (TCRs) are becoming highly attractive because of their potential to target virtually all cellular proteins, including cancer-specific epitopes, via the recognition of peptide-human leukocyte antigen (pHLA) complexes presented at the cell surface. However, because natural TCRs generally recognize cancer-derived pHLAs with very weak affinities, efforts have been made to enhance their binding strength, in some cases by several million-fold. In this study, we investigated the mechanisms underpinning human TCR affinity enhancement by comparing the crystal structures of engineered enhanced affinity TCRs with those of their wild-type progenitors. Additionally, we performed molecular dynamics simulations to better understand the energetic mechanisms driving the affinity enhancements. These data demonstrate that supra-physiological binding affinities can be achieved without altering native TCR-pHLA binding modes via relatively subtle modifications to the interface contacts, often driven through the addition of buried hydrophobic residues. Individual energetic components of the TCR-pHLA interaction governing affinity enhancements were distinct and highly variable for each TCR, often resulting from additive, or knock-on, effects beyond the mutated residues. This comprehensive analysis of affinity-enhanced TCRs has important implications for the future rational design of engineered TCRs as efficacious and safe drugs for cancer treatment.

EnvironmentAsia, 13, Special, 81-86

This thesis provides a full academic biography of the three northern major-generals appointed by government in 1655 to implement security and reform in the northern counties during the Protectorate of Oliver Cromwell, namely Charles Howard (responsible for Cumberland, Northumberland and Westmorland); Robert Lilburne (responsible for Durham and Yorkshire); and, Charles Worsley (responsible for Cheshire, Lancashire and Staffordshire). The thesis demonstrates how each of the three individuals operated their own distinctive local agendas, resulting in unique outcomes within the localities for which they were responsible. The thesis shows how these local agendas modified government policy, limiting its impact within the localities. The introductory chapter explains the historical context, highlighting how the major-generals’ regime was created in 1655 as a result of concerns of regime change as a consequence of royalist conspiracy and providential fear of God’s judgement on the nation. Chapter 1 provides a detailed analysis of the work of Charles Howard, demonstrating how his activities as major-general were shaped by his aristocratic background and power in the northern counties; attributes which made him indispensable to Cromwell in controlling the contentious Scottish border region. Chapter 2 considers Robert Lilburne, demonstrating how his radical Baptist religious beliefs, his links with powerful northern Parliamentarians and his effectiveness as a military figure combined to define his role as major-general in which he sought to reform communities and punish royalist insurgents. Chapter 3 examines Charles Worsley, showing that his position and standing emanated from Cromwell’s patronage and that he could only apply his strongly millenarian religious beliefs in north-west English communities through his role as a state agent. The thesis concludes that each of the three northern major-generals operated their own distinctive local agendas based on their unique backgrounds and the situations in which they operated. The thesis demonstrates how these local agendas had a significant effect in modifying and limiting Cromwellian policy within localities.

The ubiquitin specific protease 11 (USP11) is implicated in DNA repair, viral RNA replication, and TGF? signaling. We report the first characterization of the USP11 domain architecture and its role in regulating the enzymatic activity. USP11 consists of an N-terminal "domain present in USPs" (DUSP) and "ubiquitin-like" (UBL) domain, together referred to as DU domains, and the catalytic domain harboring a second UBL domain. Crystal structures of the DU domains show a tandem arrangement with a shortened ?-hairpin at the two-domain interface and altered surface characteristics compared to the homologues USP4 and USP15. A conserved VEVY motif is a signature feature at the two-domain interface that shapes a potential protein interaction site. Small angle X-ray scattering and gel filtration experiments are consistent with the USP11DU domains and full-length USP11 being monomeric. Unexpectedly, we reveal, through kinetic assays of a series of deletion mutants, that the catalytic activity of USP11 is not regulated through intramolecular autoinhibition or activation by the N-terminal DU or UBL domains. Moreover, ubiquitin chain cleavage assays with all eight linkages reveal a preference for Lys(63)-, Lys(6)-, Lys(33)-, and Lys(11)-linked chains over Lys(27)-, Lys(29)-, and Lys(48)-linked and linear chains consistent with USP11's function in DNA repair pathways that is mediated by the protease domain. Our data support a model whereby USP11 domains outside the catalytic core domain serve as protein interaction or trafficking modules rather than a direct regulatory function of the proteolytic activity. This highlights the diversity of USPs in substrate recognition and regulation of ubiquitin deconjugation.

VCU Theses and Dissertations

Chapter Three discusses the dream vision of Book I of the Vox Clamantis; it shows how Gower repeats the commonplaces of medieval didactic writers, regarding the peasant insurrection of 1381 as an outbreak of demonic derangement. It is seen that Gower makes use of the 'organic analogy' of society to show this madness as an infection of the entire social body. The sufferings of the nobility at the hands of the rioting mobs are described sympathetically in terms of 'grief-madness'. Thus Gower presents two very different, class-based, attitudes towards insanity. The discussion of Chaucer's Miller's and Summoner's tales in the following chapter continues the investigation of the link between madness and social class. Here it is seen how Chaucer undermines the traditional theological interpretation of madness as a punishment for sin by encouraging comparison and contrast of the many allegations of insanity in the texts. A rather different approach is taken in Chapter Five, which examines the major works of the civil servant Thomas Hoccleve. Far from regarding madness as essentially spectacular, the apparently insane narrator of Hoccleve's major poems stresses that insanity is a hidden and undetectable affliction. This conclusion, it is argued, contradicts the standard view of psychiatric history regarding madness in the Middle Ages. The relationship between madness, expressions of interiority and medieval autobiography is considered. The final chapter explores the association of madness, female unruliness and mystical rapture in The Book of Margery Kempe. It argues that the Book displays two contradictory attitudes towards madness. Kempe is eager to present madness as a moral abomination and she frequently invokes ecclesiastical authority to do so. Nevertheless, she herself is held mad by many of her contemporaries on account of her controversial devotional behaviour; this explains why madness is presented positively elsewhere in the Book, as a blessed condition of increased spiritual insight. In this sense the Book contains a craftily double-edged attempt by Kempe to vindicate her conduct.

Much can be learned from community-based environmental protection efforts in New England. The region's rural character, mixed ownerships, and resource-dependent communities provide a wealth of innovative examples of merging diverse interests toward a common goal. In many cases the key to success is listening to local constituents and interests and fostering local trust and mutual respect. In Vermont, this has most often been realized through public/ partnerships on the local level.

Bibliography: p. 142-152.