ROLE OF ICAM-1-MEDIATED ENDOCYTOSIS IN ENDOTHELIAL FUNCTION AND IMPLICATIONS FOR CARRIER-ASSISTED DRUG DELIVERY

Intercellular adhesion molecule 1 (ICAM-1) is a transmembrane protein found on the surface of vascular endothelial cells (ECs). Its expression is upregulated at inflammatory sites, allowing for targeted delivery of therapeutics using ICAM-1-binding drug carriers. Engagement of multiple copies of ICAM-1 by these drug carriers induces cell adhesion molecule (CAM)-mediated endocytosis, which results in trafficking of carriers to lysosomes and across ECs. Knowledge about the regulation behind CAM-mediated endocytosis can help improve drug delivery, but questions remain about these regulatory mechanisms. Furthermore, little is known about the natural function of this endocytic pathway. To address these gaps in knowledge, we focused on two natural binding partners of ICAM-1 that potentially elicit CAM-mediated endocytosis: leukocytes (which bind ICAM-1 via β₂ integrins) and fibrin polymers (a main component of blood clots which binds ICAM-1 via the γ3 sequence). First, inspired by properties of these natural binding partners, we varied the size and targeting moiety of model drug carriers to determine how these parameters affect CAM-mediated endocytosis. Increasing ICAM-1-targeted carrier size slowed carrier uptake kinetics, reduced carrier trafficking to lysosomes, and increased carrier transport across ECs. Changing targeting moieties from antibodies to peptides decreased particle binding and uptake, lowered trafficking to lysosomes, and increased transport across ECs. Second, using cell culture models of leukocyte/EC interactions, inhibiting regulatory elements of the CAM-mediated pathway disrupted leukocyte sampling, a process crucial to leukocyte crossing of endothelial layers (transmigration). This inhibition also decreased leukocyte transmigration across ECs, specifically through the transcellular route, which occurs through a single EC without disassembly of cell-cell junctions. Third, fibrin meshes, which mimic blood clot fragments/remnants, bound to ECs at ICAM-1-enriched sites and were internalized by the endothelium. Inhibiting the CAM-mediated pathway disrupted this uptake. Following endocytosis, fibrin meshes trafficked to lysosomes where they were degraded. In mouse models, CAM-mediated endocytosis of fibrin meshes appeared to remove fibrin remnants at the endothelial surface, preventing re-initiation of the coagulation cascade. Overall, these results support a link between CAM-mediated endocytosis and leukocyte transmigration as well as uptake of fibrin materials by ECs. Furthermore, these results will guide the future design of ICAM-1-targeted carrier-assisted therapies.

Playing for Real: Designing Alternate Reality Games in Learning Contexts

Alternate Reality Game (ARG) represent a new genre of transmedia practice where players hunt for scattered clues, make sense of disparate information, and solve puzzles to advance an ever-evolving storyline. Players participate in ARGs using multiple communications technologies, ranging from print materials to mobile devices. However, many interaction design challenges must be addressed to weave these everyday communication tools together into an immersive, participatory experience. Transmedia design is not an everyday process. Designers must create and connect story bits across multiple media (video, audio, text) and multiple platforms (phones, computers, physical spaces). Furthermore, they must engage with players of varying skill levels. Few studies to-date have explored the design process of ARGs in learning contexts. Fewer still have focused on challenges involved in designing for youth (13-17 years old). In this study, I explore the process of designing ARGs as vehicles for promoting information literacy and participatory culture for adolescents (13-17 years old). Two ARG design scenarios, distinguished by target learning environment (formal and informal context) and target audience (adolescents), comprise the two cases that I examine. Through my analysis of these two design cases, I articulate several unique challenges faced by designers who create interactive, transmedia stories for – and with – youth. Drawing from these design challenges, I derive a repertoire of design strategies that future designers and researchers may use to create and implement ARGs for teens in learning contexts. In particular, I propose a narrative design framework that allows for the categorization of ARGs as storytelling constructs that lie along a continuum of participation and interaction. The framework can serve as an analytic tool for researchers and a guide for designers. In addition, I establish a framework of social roles that designers may employ to craft transmedia narratives before live launch and to promote and scaffold player participation after play begins. Overall, the contributions of my study include theoretical insights that may advance our understanding of narrative design and analysis as well as more practical design implications for designers and practitioners seeking to incorporate transmedia features into learning experiences that target youth.

Topics in equilibrium and nonequilibrium thermodynamics: computing crystalline free energies and engineering Maxwell’s demon.

This dissertation covers two separate topics in statistical physics. The first part of the dissertation focuses on computational methods of obtaining the free energies (or partition functions) of crystalline solids. We describe a method to compute the Helmholtz free energy of a crystalline solid by direct evaluation of the partition function. In the many-dimensional conformation space of all possible arrangements of N particles inside a periodic box, the energy landscape consists of localized islands corresponding to different solid phases. Calculating the partition function for a specific phase involves integrating over the corresponding island. Introducing a natural order parameter that quantifies the net displacement of particles from lattices sites, we write the partition function in terms of a one-dimensional integral along the order parameter, and evaluate this integral using umbrella sampling. We validate the method by computing free energies of both face-centered cubic (FCC) and hexagonal close-packed (HCP) hard sphere crystals with a precision of $10^{-5}k_BT$ per particle. In developing the numerical method, we find several scaling properties of crystalline solids in the thermodynamic limit. Using these scaling properties, we derive an explicit asymptotic formula for the free energy per particle in the thermodynamic limit. In addition, we describe several changes of coordinates that can be used to separate internal degrees of freedom from external, translational degrees of freedom. The second part of the dissertation focuses on engineering idealized physical devices that work as Maxwell's demon. We describe two autonomous mechanical devices that extract energy from a single heat bath and convert it into work, while writing information onto memory registers. Additionally, both devices can operate as Landauer's eraser, namely they can erase information from a memory register, while energy is dissipated into the heat bath. The phase diagrams and the efficiencies of the two models are solved and analyzed. These two models provide concrete physical illustrations of the thermodynamic consequences of information processing.

SELECTED VARIATIONS BY COMPOSERS IN GERMANY AND FRANCE

The Haward of Dictionary of Music (1983), defines variation as "a technique modifying a given musical idea." From the Baroque period on, the form and the techniques of variation were developed and enriched in Germany and France. Therefore, I presented the works of composers from these two nations. Even though there was a vast number of possibilities, I wanted to be scholastically fair and interesting in making my selections by choosing well-known pieces along with lesser-known ones. Haydn's well-known Variations in F minor consist of two sets of double variations which break into an improvisation fantasy. The first movement of Beethoven Sonata in A flat major, Op. 26, is a set of five variations on the composer's original theme. The variations are positioned in the first movement instead of Sonata-Allegro form. In 1861 Brahrns composed the Variations and Fugue, Op. 24, on the theme of Handel. Brahms displays a wealth of rhythmic, harmonic and textural contrasts in the variations. Chopin's E Major Variations without opus number are written on a Swiss influenced German folksong. Faure's Theme and Variations in C sharp minor, Op. 73, includes eleven variations. The work displays the composer's subtlety, grace and reticence. 12 Variationen iiber ein eigenes Thema were written by Alban Berg as a composition study with Schonberg. The Finale of Dutilleux's Piano Sonata, titled "Chorale with Variations", is written in an impressionistic style. A rich expressiveness is well blended in a classical form. In 1742, the remarkable Aria and thirty variations known as the Goldberg Variations were composed by J. S. Bach. The thirty Variations are unified by the bass line, which forms the foundation of the Aria. The pieces discussed above were presented in three recitals. Compact disc recordings of these recitals are available in the Michelle Smith Performing Arts Library of the Clarice Smith Performing Arts Center at the University of Maryland.

The Scherzo in Selected Works for Piano

This performance project will cover performing issues in terms of technique in the scherzo. The Dictionary of Musical Terms defines technique as "the system of creating music, the musical slull to show personality by controlling tones that is not an abstract theory but a practical ability in composition or performance." My project focuses on techniques in fast tempos, specifically those found in the scherzo form and in concertos containing a scherzo character. The term scherzo has varied in its meaning and form throughout history. In the Baroque period, a scherzo was a work of light vocal or instrumental character. In the Classical period, scherzo still meant light in style, but it also indicated a quick tempo, often in 2/4 time. The scherzo was usually a single movement in a suite or multi-movement work. Like the minuet form, the scherzo contained a contrasting trio section. The scherzo was also standard in Romantic and post-Romantic symphonies and related genres. Because of the high degree of subjectivity in Romantic music, genres that stressed emotional content over abstract form developed rapidly. Some composers even wrote one-movement pieces entitled scherzo. These pieces became very important because they usually expressed a particular character or mood. The objective of my dissertation project is to research scherzo-like concertos, scherzo as single movements in larger forms, and scherzo as independent works. My first recital will consist of two concertos with a scherzo-like character. These are Mozart's Piano Concerto No. 9 i ?nl Major; K. 271 and Ravel's Piano Concerto in G Major. I will perform these works in December 2002 with a second piano. In addition, I will perform the Ravel with an orchestra in 2003. My second recital will consist of two parts. The fxst half presents multi-movement works with scherzo movements. The pieces are Haydn's Piano Sonata No. 3 in F Majol; Hob. WI/9, Beethoven's Piano Sonata No. 10 in G Major; Op. 14, No. 2. The second half presents independent four scherzi by Chopin. The final program will also include multi-movement works containing scherzo and independent scherzo. These are Prokofiev's Piano Sonata No. 2 in D minor, Op. 14, Grieg Lyric Pieces Op. 54, Schubert Zwei Scherzi D. 593 and Copland Scherzo humoristique; Le Chat et la Souris (The Cat and the Mouse).

USE OF VARIATION AND FUGUE IN PIANO MUSIC FOLLOWING THE CLASSICAL PERIOD

The variation and fugue originated from the 15th and 16th centuries and blossomed during the Baroque and Classical Periods. In a variation, a theme with a particular structure precedes a series of pieces that usually have the same or very similar structure. A fugue is a work written in imitative counterpoint in which the theme is stated successively in all voices of polyphonic texture. Beethoven’s use of variation and fugue in large scale works greatly influenced his contemporaries. After the Classical Period, variations continued to be popular, and numerous composers employed the technique in various musical genres. Fugues had pedagogical associations, and by the middle of 19th century became a requirement in conservatory instruction, modeled after Bach’s Well-Tempered Clavier. In the 20th century, the fugue was revived in the spirit of neoclassicism; it was incorporated in sonatas, and sets of preludes and fugues were composed. Schubert's Wanderer Fantasy presents his song Der Wanderer through thematic transformations, including a fugue and a set of variations. Liszt was highly influenced by this, as shown in his thematic transformations and the fugue as one of the transformations in his Sonata in b. In Schumann’s Symphonic Études, Rachmaninoff's Rhapsody on a Theme of Paganini and Copland’s Piano Variations, the variation serves as the basis for the entire work. Prokofiev and Schubert take a different approach in Piano Concerto No. 3 and Wanderer Fantasy, employing the variation in a single movement. Unlike Schubert and Liszt's use of the fugue as a part of the piece or movement, Franck’s Prelude Chorale et Fugue and Shchedrin’s Polyphonic Notebook use it in its independent form. Since the Classical Period, the variation and fugue have evolved from stylistic and technical influences of earlier composers. It is interesting and remarkable to observe the unique effects each had on a particular work. As true and dependable classic forms, they remain popular by offering the composer an organizational framework for musical imagination.