An active learning approach for rapid characterization of endothelial cells in human tumors.

Currently, no available pathological or molecular measures of tumor angiogenesis predict response to antiangiogenic therapies used in clinical practice. Recognizing that tumor endothelial cells (EC) and EC activation and survival signaling are the direct targets of these therapies, we sought to develop an automated platform for quantifying activity of critical signaling pathways and other biological events in EC of patient tumors by histopathology. Computer image analysis of EC in highly heterogeneous human tumors by a statistical classifier trained using examples selected by human experts performed poorly due to subjectivity and selection bias. We hypothesized that the analysis can be optimized by a more active process to aid experts in identifying informative training examples. To test this hypothesis, we incorporated a novel active learning (AL) algorithm into FARSIGHT image analysis software that aids the expert by seeking out informative examples for the operator to label. The resulting FARSIGHT-AL system identified EC with specificity and sensitivity consistently greater than 0.9 and outperformed traditional supervised classification algorithms. The system modeled individual operator preferences and generated reproducible results. Using the results of EC classification, we also quantified proliferation (Ki67) and activity in important signal transduction pathways (MAP kinase, STAT3) in immunostained human clear cell renal cell carcinoma and other tumors. FARSIGHT-AL enables characterization of EC in conventionally preserved human tumors in a more automated process suitable for testing and validating in clinical trials. The results of our study support a unique opportunity for quantifying angiogenesis in a manner that can now be tested for its ability to identify novel predictive and response biomarkers.

Communications inspired linear discriminant analysis

We study the problem of supervised linear dimensionality reduction, taking an information-theoretic viewpoint. The linear projection matrix is designed by maximizing the mutual information between the projected signal and the class label. By harnessing a recent theoretical result on the gradient of mutual information, the above optimization problem can be solved directly using gradient descent, without requiring simplification of the objective function. Theoretical analysis and empirical comparison are made between the proposed method and two closely related methods, and comparisons are also made with a method in which Rényi entropy is used to define the mutual information (in this case the gradient may be computed simply, under a special parameter setting). Relative to these alternative approaches, the proposed method achieves promising results on real datasets. Copyright 2012 by the author(s)/owner(s).

Visual Disobedience: The Geopolitics of Experimental Art in Central America, 1990-Present

This dissertation centers on the relationship between art and politics in postwar Central America as materialized in the specific issues of racial and gendered violence that derive from the region's geopolitical location and history. It argues that the decade of the 1990s marks a moment of change in the region's cultural infrastructure, both institutionally and conceptually, in which artists seek a new visual language of experimental art practices to articulate and conceptualize a critical understanding of place, experience and knowledge. It posits that visual and conceptual manifestations of violence in Central American performance, conceptual art and installation extend beyond a critique of the state, and beyond the scope of political parties in perpetuating violent circumstances in these countries. It argues that instead artists use experimental practices in art to locate manifestations of racial violence in an historical system of domination and as a legacy of colonialism still witnessed, lived, and learned by multiple subjectivities in the region. In this postwar period artists move beyond the cold-war rhetoric of the previous decades and instead root the current social and political injustices in what Aníbal Quijano calls the `coloniality of power.' Through an engagement of decolonial methodologies, this dissertation challenges the label "political art" in Central America and offers what I call "visual disobedience" as a response to the coloniality of seeing. I posit that visual colonization is yet another aspect of the coloniality of power and indispensable to projects of decolonization. It offers an analysis of various works to show how visual disobedience responds specifically to racial and gender violence and the equally violent colonization of visuality in Mesoamerica. Such geopolitical critiques through art unmask themes specific to life and identity in contemporary Central America, from indigenous genocide, femicide, transnational gangs, to mass imprisonments and a new wave of social cleansing. I propose that Central American artists--beyond an anti-colonial stance--are engaging in visual disobedience so as to construct decolonial epistemologies in art, through art, and as art as decolonial gestures for healing.

Exploring Musical Diversity in the Collaborative Repertoire from 1880 through 1963

It is essential in musical performance not only to convey the unique language of the composers but also to approach each composition from the perspective of its style. During the 20th century, diverse musical idioms co-existed, sometimes mixing or fusing, yet retaining recognizable characteristics and thereby remaining distinctive. This dissertation explores myriad examples from Late Romanticism/Post- Romanticism, Naturalism, Neo-Classicism, Nationalism and Impressionism composed during this unusually rich period. In order to explore a broad range of collaborative repertoire and to deepen my knowledge of the styles and performance practices relating to these pieces, I studied and performed the repertoire with pianist Eunae Baik–Kim, clarinetist Jihoon Chang, and singers Joshua Brown and Young Joo Lee. The first program featured Post-Romantic, Neo-Classic and Impressionist two-piano works composed by Debussy, Rachmaninoff, and Stravinsky. Each of the three composers used their own distinctive harmonies, rhythms, melodic inventions, pedaling and figurations. In all of the works, both piano parts were densely interwoven, having equal importance. Lied and operatic aria was the focus of the second recital. Brahms’ Vier Ernste Gesänge Op. 121, Ravel’s Don Quichotte a Dulcineé and Italian, French and German operatic arias were the examples of Post-Romanticism and Nationalism. The representative composers were Verdi, Massenet, Korngold, Leoncavallo, Ravel and Wagner. Despite the fact that all of the repertoire was written in traditional musical forms, the composers’ unique voices mark each work as belonging to a particular genre. The third recital focused on Post-Romantic and Impressionistic music written for clarinet and piano: the Première Rhapsodie by Debussy, the Sonata by Poulenc and Brahms’ Sonata in F minor Op. 120, No. 1. These works, although profoundly different in style, share elements of simplicity, clarity and elegance as well as technical virtuosity, articulation and profound musical depth. The three recitals which comprise this dissertation project were performed at the University of Maryland Gildenhorn Recital Hall on February 27, 2010, October 25, 2010, and January 31, 2011. The recitals were recorded on compact disc and are archived within the Digital Repository at the University of Maryland (DRUM).

VINTAGE MODERNISM: EARLY EXPERIMENTALIST MUSIC FOR VIOLIN IN THE UNITED STATES

The historical narrative of concert music in the early twentieth century has focused a great deal on the influence of European composers as well as the American composers who went to Europe to study. Often overlooked, however, is the influence of an entire generation of composers working in the United States during that time period. These artists experimented with polyrhythm, polytonality, dissonant counterpoint, and a whole host of other musical techniques in order to express their perceptions of a changing world. Over the years, the new techniques became associated with various movements including futurism, experimentalism and ultramodernism. Regardless of label, these composers were some of the first to introduce the new musical styles to the listening public.The recitals that make up this dissertation explore the sound world of experimentalist composers working in the United States during the early twentieth century. Serving as the foundation of these recitals are all four of the violin sonatas by Charles Ives, the “grandfather” of modernist music whose financial support helped to foster a whole generation of American composers. Also prominently featured is the music of Henry Cowell. His Suite for Violin and Piano, Mosaic Quartet (String Quartet No. 3), and Quartet Euphometric demonstrate the composer's use of cluster tones, dissonant counterpoint, polymeter, and indeterminate form. Additional works by George Antheil, Leo Ornstein, Wallingford Riegger, Dane Rudhyar, Carl Ruggles, and Ruth Crawford (Seeger) highlight other approaches taken by members of the ultra-modernist movement. Rounding out the repertoire for these recitals are works by Johanna Beyer and Conlon Nancarrow, both of whom either worked with or were influenced by Cowell in some way. All of the pieces selected date roughly from 1900 to the mid 1930's. Thus, the purpose of these recitals is not to provide a comprehensive overview of each composer's development, but rather to examine the influence and interconnections of a specific cross-section of the musical landscape.

The Representation of Emotion in Autonomic and Central Nervous System Activity

Phenomenologically, humans effectively label and report feeling distinct emotions, yet the extent to which emotions are represented categorically in nervous system activity is controversial. Theoretical accounts differ in this regard, some positing distinct emotional experiences emerge from a dimensional representation (e.g., along axes of valence and arousal) whereas others propose emotions are natural categories, with dedicated neural bases and associated response profiles. This dissertation aims to empirically assess these theoretical accounts by examining how emotions are represented (either as disjoint categories or as points along continuous dimensions) in autonomic and central nervous system activity by integrating psychophysiological recording and functional neuroimaging with machine-learning based analytical methods. Results demonstrate that experientially, emotional events are well-characterized both along dimensional and categorical frameworks. Measures of central and peripheral responding discriminate among emotion categories, but are largely independent of valence and arousal. These findings suggest dimensional and categorical aspects of emotional experience are driven by separable neural substrates and demonstrate that emotional states can be objectively quantified on the basis of nervous system activity.

Chlamydia trachomatis Infection Leads to Defined Alterations to the Lipid Droplet Proteome in Epithelial Cells.

The obligate intracellular bacterium Chlamydia trachomatis is a major human pathogen and a main cause of genital and ocular diseases. During its intracellular cycle, C. trachomatis replicates inside a membrane-bound vacuole termed an "inclusion". Acquisition of lipids (and other nutrients) from the host cell is a critical step in chlamydial replication. Lipid droplets (LD) are ubiquitous, ER-derived neutral lipid-rich storage organelles surrounded by a phospholipids monolayer and associated proteins. Previous studies have shown that LDs accumulate at the periphery of, and eventually translocate into, the chlamydial inclusion. These observations point out to Chlamydia-mediated manipulation of LDs in infected cells, which may impact the function and thereby the protein composition of these organelles. By means of a label-free quantitative mass spectrometry approach we found that the LD proteome is modified in the context of C. trachomatis infection. We determined that LDs isolated from C. trachomatis-infected cells were enriched in proteins related to lipid metabolism, biosynthesis and LD-specific functions. Interestingly, consistent with the observation that LDs intimately associate with the inclusion, a subset of inclusion membrane proteins co-purified with LD protein extracts. Finally, genetic ablation of LDs negatively affected generation of C. trachomatis infectious progeny, consistent with a role for LD biogenesis in optimal chlamydial growth.

D-Amino acid peptide residualizing agents bearing N-hydroxysuccinimido- and maleimido-functional groups and their application for trastuzumab radioiodination.

INTRODUCTION: Proteins that undergo receptor-mediated endocytosis are subject to lysosomal degradation, requiring radioiodination methods that minimize loss of radioactivity from tumor cells after this process occurs. To accomplish this, we developed the residualizing radioiodination agent N(ϵ)-(3-[(*)I]iodobenzoyl)-Lys(5)-N(α)-maleimido-Gly(1)-D-GEEEK (Mal-D-GEEEK-[(*)I]IB), which enhanced tumor uptake but also increased kidney activity and necessitates generation of sulfhydryl moieties on the protein. The purpose of the current study was to synthesize and evaluate a new D-amino acid based agent that might avoid these potential problems. METHODS: N(α)-(3-iodobenzoyl)-(5-succinimidyloxycarbonyl)-D-EEEG (NHS-IB-D-EEEG), which contains 3 D-glutamates to provide negative charge and a N-hydroxysuccinimide function to permit conjugation to unmodified proteins, and the corresponding tin precursor were produced by solid phase peptide synthesis and subsequent conjugation with appropriate reagents. Radioiodination of the anti-HER2 antibody trastuzumab using NHS-IB-D-EEEG and Mal-D-GEEEK-IB was compared. Paired-label internalization assays on BT474 breast carcinoma cells and biodistribution studies in athymic mice bearing BT474M1 xenografts were performed to evaluate the two radioiodinated D-peptide trastuzumab conjugates. RESULTS: NHS-[(131)I]IB-D-EEEG was produced in 53.8%±13.4% and conjugated to trastuzumab in 39.5%±7.6% yield. Paired-label internalization assays with trastuzumab-NHS-[(131)I]IB-D-EEEG and trastuzumab-Mal-D-GEEEK-[(125)I]IB demonstrated similar intracellular trapping for both conjugates at 1h ((131)I, 84.4%±6.1%; (125)I, 88.6%±5.2%) through 24h ((131)I, 60.7%±6.8%; (125)I, 64.9%±6.9%). In the biodistribution experiment, tumor uptake peaked at 48 h (trastuzumab-NHS-[(131)I]IB-D-EEEG, 29.8%±3.6%ID/g; trastuzumab-Mal-D-GEEEK-[(125)I]IB, 45.3%±5.3%ID/g) and was significantly higher for (125)I at all time points. In general, normal tissue levels were lower for trastuzumab-NHS-[(131)I]IB-D-EEEG, with the differences being greatest in kidneys ((131)I, 2.2%±0.4%ID/g; (125)I, 16.9%±2.8%ID/g at 144 h). CONCLUSION: NHS-[(131)I]IB-D-EEEG warrants further evaluation as a residualizing radioiodination agent for labeling internalizing antibodies/fragments, particularly for applications where excessive renal accumulation could be problematic.

Adjunctive albuterol enhances the response to enzyme replacement therapy in late-onset Pompe disease.

Effective dosages for enzyme replacement therapy (ERT) in Pompe disease are much higher than for other lysosomal storage disorders, which has been attributed to low cation-independent mannose-6-phosphate receptor (CI-MPR) in skeletal muscle. We have previously demonstrated the benefit of increased CI-MPR-mediated uptake of recombinant human acid-α-glucosidase during ERT in mice with Pompe disease following addition of albuterol therapy. Currently we have completed a pilot study of albuterol in patients with late-onset Pompe disease already on ERT for >2 yr, who were not improving further. The 6-min walk test (6MWT) distance increased in all 7 subjects at wk 6 (30±13 m; P=0.002), wk 12 (34±14 m; P=0.004), and wk 24 (42±37 m; P=0.02), in comparison with baseline. Grip strength was improved significantly for both hands at wk 12. Furthermore, individual subjects reported benefits; e.g., a female patient could stand up from sitting on the floor much more easily (time for supine to standing position decreased from 30 to 11 s), and a male patient could readily swing his legs out of his van seat (hip abduction increased from 1 to 2+ on manual muscle testing). Finally, analysis of the quadriceps biopsies suggested increased CI-MPR at wk 12 (P=0.08), compared with baseline. With the exception of 1 patient who succumbed to respiratory complications of Pompe disease in the first week, only mild adverse events have been reported, including tremor, transient difficulty falling asleep, and mild urinary retention (requiring early morning voiding). Therefore, this pilot study revealed initial safety and efficacy in an open label study of adjunctive albuterol therapy in patients with late-onset Pompe disease who had been stable on ERT with no improvements noted over the previous several years.

A scalable strategy for the parallelization of multiphysics unstructured mesh-iterative codes on distributed-memory systems

Realizing scalable performance on high performance computing systems is not straightforward for single-phenomenon codes (such as computational fluid dynamics [CFD]). This task is magnified considerably when the target software involves the interactions of a range of phenomena that have distinctive solution procedures involving different discretization methods. The problems of addressing the key issues of retaining data integrity and the ordering of the calculation procedures are significant. A strategy for parallelizing this multiphysics family of codes is described for software exploiting finite-volume discretization methods on unstructured meshes using iterative solution procedures. A mesh partitioning-based SPMD approach is used. However, since different variables use distinct discretization schemes, this means that distinct partitions are required; techniques for addressing this issue are described using the mesh-partitioning tool, JOSTLE. In this contribution, the strategy is tested for a variety of test cases under a wide range of conditions (e.g., problem size, number of processors, asynchronous / synchronous communications, etc.) using a variety of strategies for mapping the mesh partition onto the processor topology.

SMARTFIRE: an integrated computational fluid dynamics code and expert system for fire field modelling

SMARTFIRE, an open architecture integrated CFD code and knowledge based system attempts to make fire field modeling accessible to non-experts in Computational Fluid Dynamics (CFD) such as fire fighters, architects and fire safety engineers. This is achieved by embedding expert knowledge into CFD software. This enables the 'black-art' associated with the CFD analysis such as selection of solvers, relaxation parameters, convergence criteria, time steps, grid and boundary condition specification to be guided by expert advice from the software. The user is however given the option of overriding these decisions, thus retaining ultimate control. SMARTFIRE also makes use of recent developments in CFD technology such as unstructured meshes and group solvers in order to make the CFD analysis more efficient. This paper describes the incorporation within SMARTFIRE of the expert fire modeling knowledge required for automatic problem setup and mesh generation as well as the concept and use of group solvers for automatic and manual dynamic control of the CFD code.

Constructive trusts, illegal purpose and locus poenitentiae

Analyses the Family Division decision in Q v Q on the competing claims of a father and son to beneficial ownership of a house in which the father sought to on a secret agreement entered into with his sons when transferring the property into their joint names, intended to subvert the inheritance tax rules on lifetime gifts by retaining the right to have the property transferred back to him, and the son relied on a later agreement with his brother to transfer the house into his sole name, in reliance on which he and his wife had acted to their detriment by paying for its upkeep and renovation.

Meaning of "residing with" and succession rights

Reviews case law on the meaning of the phrase "residing with" as used in the Housing Act 1985 s.87 to identify one of the qualifying requirements for the right to succeed to a secure tenancy. Focuses on the Court of Appeal decision in Freeman v Islington LBC on whether a daughter intended to have her settled home with her father where, in the year preceding her father's death she lived with him full time in order to care for him whilst retaining her own flat and using it as her correspondence address for everything but her credit card. Lists conclusions that can be drawn from the case law.

Personal and proprietary tenancies

Examines the Chancery Division ruling in London Development Agency v Nidai on whether a number of agreements providing for the construction of a bridge and shop premises on the retaining walls of a river resulted in a binding legal lease or a series of bare licences. Comments on the failure of the judgment to mention the House of Lords ruling in Bruton v London & Quadrant Housing Trust and discusses whether a Bruton tenancy is capable of binding third parties.

On treating the symptoms and not the cause: reflections on the Dangerous Dogs Act

The experience of saving a dog that later turned out to be a Pit Bull and therefore banned under the Dangerous Dogs Act 1991, made me investigate the Act and its implications. The Act is not built on evidence and by compiling results from different studies on dog bites and breed‐specific legislation in different countries the conclusion is that there is not much empirical support for breed bans either. ‘Dangerous breeds’ do not bite more frequently than German Shepherds and directing legislation towards certain breeds deemed as ‘dangerous’ cannot therefore be seen as justified. The strength of the label ‘dangerous dog’ seems to rule out policies that follow the facts and there is more treating of symptoms than causes. [From the Author]