No / The orientation-sensitive performance of the Schlumberger array, when used to survey narrow, linear features, has long been recognized in geophysical prospecting for geology. However, in spite of frequent use of the array for archaeological survey, particularly in eastern Europe and the Far East, this directional effect is not apparent in the survey of walls and ditches. In order to examine the array's performance some experiments were carried out in a shallow electrolytic tank using insulating and conducting cylinders. Broadside and longitudinal traverses with systematic expansion of the current electrode spacing facilitated the production of pseudosections. The results confirmed the high selectivity of the Schlumberger response to the orientation of the feature. Broadside traverse of the conductor and longitudinal traverse of the insulator produced very large changes: much smaller signals were recorded for the alternative orientations. A subsequent experiment, however, on a simulated ditch in bedrock revealed no signal. The directional effect for a linear insulator was confirmed in field studies of a simple stone-walled structure. Implications for survey of low-contrast linear archaeological features are discussed.

Yes / A key question for temporal processing research is how the nervous system extracts event duration, despite a notable lack of neural structures dedicated to duration encoding. This is in stark contrast with the orderly arrangement of neurons tasked with spatial processing. In this study, we examine the linkage between the spatial and temporal domains. We use sensory adaptation techniques to generate after-effects where perceived duration is either compressed or expanded in the opposite direction to the adapting stimulus’ duration. Our results indicate that these after-effects are broadly tuned, extending over an area approximately five times the size of the stimulus. This region is directly related to the size of the adapting stimulus—the larger the adapting stimulus the greater the spatial spread of the aftereffect. We construct a simple model to test predictions based on overlapping adapted versus non-adapted neuronal populations and show that our effects cannot be explained by any single, fixed-scale neural filtering. Rather, our effects are best explained by a self-scaled mechanism underpinned by duration selective neurons that also pool spatial information across earlier stages of visual processing. / J.H. is supported by the Vision Research Trust (43069). N.W.R. is supported by a Wellcome Trust Research Career Development Fellowship (WT097387).

Natural sound, such as conspecific vocalizations and human speech, represents an important part of the sensory signals animals and humans encounter in their daily lives. This dissertation investigates the neural mechanisms involved in creating response selectivity for complex features of natural acoustic signals and demonstrates that selectivity for spectral motion cues provides a neural mechanism to encode communication signals in the auditory midbrain. Spectral motion is defined as the movement of sound energy upward or downward in frequency at a certain velocity, and is believed to provide the auditory system with an important perceptual cue in the processing of human speech. Using the Mexican free-tailed bat, tadarida brasiliensis, as a model system, this research examined the role of selectivity for spectral motion cues, such as direction and velocity, in creating response selectivity for specific features of the social communication signals emitted by these animals. We show that auditory neurons in the midbrain nucleus of the inferior colliculus (IC) are specifically tuned for the frequency-modulated (FM) direction and velocities found in their conspecific vocalizations. This close agreement between neural tuning and features of natural conspecific signals shows that auditory neurons have evolved to specifically encode features of signals that are vital for the survival of the animal. Furthermore, we find that the neural computations resulting in selectivity for spectral motion are analogous to mechanisms observed in selectivity for visual motion, suggesting the evolution of similar neural mechanisms across sensory modalities. / text

The Two-pore channel (TPC1-3) family represents a recently identified class of endolysosomal ion channels. TPCs were originally proposed to be promising candidate channels for NAADP-induced Ca²⁺ release. However, subsequent studies have emerged to propose an alternative view where TPCs may be Na+-selective channels regulated by the lysosome-specific phosphoinositide PI(3,5)P2 or voltage in an isoform-dependent manner. This thesis asks the question of whether pharmacological and ion permeation properties of TPCs, in particular the human TPC2, may satisfy or may be consistent with the requirement of a potential NAADP-sensitive Ca²⁺-release channel. These fundamental properties of hTPC2 were approached using patch-clamp electrophysiology and confocal fluorescence microscopy, and were analysed quantitatively to extract relevant physical parameters important to our understanding of their physiological and functional significance. Chapter 2 presents the basic electrophysiological characterisation of hTPC2. It follows a logical way by first determining the ion permeation properties, followed by the investigation of its physical relation with fractional Ca²⁺ current and Ca²⁺ nanodomains to rigorously prove that this Na⁺ selectivity is sufficient to ensure negligible Ca²⁺ leakage both experimentally and theoretically. This follows the logic that matter must not be created nor destroyed so that a Na+-selective channel that poses a physiologically significant energy barrier to Ca²⁺ permeation from one side would not lead to the creation of Ca²⁺ on the other side. Chapter 3 represents a natural progression from Chapter 2 and is aimed at investigating the underlying mechanisms responsible for the electrophysiological ion selectivity observed. This chapter also follows a logical way by first identifying spermine as a high valence intracellular blocker, its mutual antagonism with different external ionic species that allows the determination of ion-binding affinity, followed by the determination of the concentration dependence of ion conduction to identify possible lower affinity binding. By considering all the above qualities, the outcome is a coherent description and connection of ion binding selectivity, kinetic selectivity and ion binding configuration with the observed electrophysiological selectivity. Chapter 4 discusses the missing puzzles and how these questions might be addressed.

No description available.

Querying and efficiently analyzing complex data is required to gain valuable business insights, to support machine learning applications, and to make up-to-date information available. Therefore, this thesis investigates opportunities and challenges of selecting the most efficient execution strategy for analytical queries. These challenges include hard-to-capture data characteristics such as skew and correlation, the support of arbitrary data types, and the optimization time overhead of complex queries. Existing approaches often rely on optimistic assumptions about the data distribution, which can result in significant response time delays when these assumptions are not met. On the contrary, we focus on robust query optimization, emphasizing consistent query performance and applicability. Our presentation follows the general select-project-join query pattern, representing the fundamental stages of analytical query processing. To support arbitrary data types and complex filter expressions in the select stage, a novel sampling-based selectivity estimator is developed. Our approach exploits information from filter subexpressions and estimates correlations that are not captured by existing sampling-based methods. We demonstrate improved estimation accuracy and query execution time. Further, to minimize the runtime overhead of sampling, we propose new techniques that exploit access patterns and auxiliary database objects such as indices. For the join stage, we introduce a robust optimization approach by developing an upper-bound join enumeration strategy that connects accurate filter selectivity estimates –e.g., using our sampling-based approach– to join ordering. We demonstrate that join orders based on our upper-bound join ordering strategy achieve more consistent performance and faster workload execution on state-of-the-art database systems. However, besides identifying good logical join orders, it is crucial to determine appropriate physical join operators before query plan execution. To understand the importance of fine-grained physical operator selections, we exhaustively execute fixed join orders with all possible operator combinations. This analysis reveals that none of the investigated query optimizers fully reaches the potential of optimal operator decisions. Based on these insights and to achieve fine-grained operator selections for the previously determined join orders, the thesis presents a lightweight learning-based physical execution plan refinement component called. We show that this refinement component consistently outperforms existing approaches for physical operator selection while enabling a novel two-stage optimizer design. We conclude the thesis by providing a framework for the two-stage optimizer design that allows users to modify, replicate, and further analyze the concepts discussed throughout this thesis.:1 INTRODUCTION 1.1 Analytical Query Processing . . . . . . . . . . . . . . . . . . . . . . . . . . 12 1.2 Select-Project-Join Queries . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 1.3 Basics of SPJ Query Optimization . . . . . . . . . . . . . . . . . . . . . . . 14 1.3.1 Plan Enumeration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 1.3.2 Cost Model . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15 1.3.3 Cardinality Estimation . . . . . . . . . . . . . . . . . . . . . . . . . . 15 1.4 Robust SPJ Query Optimization . . . . . . . . . . . . . . . . . . . . . . . . 16 1.4.1 Tail Latency Root Cause Analysis . . . . . . . . . . . . . . . . . . . 17 1.4.2 Tenets of Robust Query Optimization . . . . . . . . . . . . . . . . . 19 1.5 Contribution . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19 1.6 Outline . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20 2 SELECT (-PROJECT) STAGE 2.1 Sampling for Selectivity Estimation . . . . . . . . . . . . . . . . . . . . . . 24 2.2 Related Work . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28 2.2.1 Combined Selectivity Estimation (CSE) . . . . . . . . . . . . . . . . 29 2.2.2 Kernel Density Estimator . . . . . . . . . . . . . . . . . . . . . . . . . 31 2.2.3 Machine Learning . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32 2.3 Beta Estimator for 0-Tuple-Situations . . . . . . . . . . . . . . . . . . . . . 33 2.3.1 Methodology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33 2.3.2 Beta Distribution in Non-0-TS . . . . . . . . . . . . . . . . . . . . . . 35 2.3.3 Parameter Estimation in 0-TS . . . . . . . . . . . . . . . . . . . . . . 37 2.3.4 Selectivity Estimation and Predicate Ordering . . . . . . . . . . . 39 2.3.5 Evaluation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46 2.4 Customized Sampling Techniques . . . . . . . . . . . . . . . . . . . . . . 53 2.4.1 Focused Sampling . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54 2.4.2 Conditional Sampling . . . . . . . . . . . . . . . . . . . . . . . . . . 56 2.4.3 Zone Pruning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58 2.4.4 Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59 2.5 Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59 3 JOIN STAGE: LOGICAL ENUMERATION 3.1 Related Work . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62 3.1.1 Point Estimates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63 3.1.2 Join Cardinality Upper Bound . . . . . . . . . . . . . . . . . . . . . 64 3.2 Upper Bound Join Enumeration with Synopsis (UES) . . . . . . . . . . . . 66 3.2.1 U-Block: Simple Upper Bound for Joins . . . . . . . . . . . . . . . . 67 3.2.2 E-Block: Customized Enumeration Scheme . . . . . . . . . . . . . 68 3.2.3 UES Algorithm . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69 3.3 Evaluation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71 3.3.1 General Performance . . . . . . . . . . . . . . . . . . . . . . . . . . 72 3.3.2 Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74 3.4 Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 76 4 JOIN STAGE: PHYSICAL OPERATOR SELECTION 4.1 Operator Selection vs Join Ordering . . . . . . . . . . . . . . . . . . . . . 77 4.2 Related Work . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 80 4.2.1 Adaptive Query Processing . . . . . . . . . . . . . . . . . . . . . . 80 4.2.2 Bandit Optimizer (Bao) . . . . . . . . . . . . . . . . . . . . . . . . . 81 4.3 TONIC: Learned Physical Join Operator Selection . . . . . . . . . . . . . 82 4.3.1 Query Execution Plan Synopsis (QEP-S) . . . . . . . . . . . . . . . 83 4.3.2 QEP-S Life-Cycle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 84 4.3.3 QEP-S Design Considerations . . . . . . . . . . . . . . . . . . . . . . 87 4.4 Evaluation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 89 4.4.1 Performance Factors . . . . . . . . . . . . . . . . . . . . . . . . . . . 90 4.4.2 Rate of Improvement . . . . . . . . . . . . . . . . . . . . . . . . . . 92 4.4.3 Data Shift . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95 4.4.4 TONIC - Runtime Traits . . . . . . . . . . . . . . . . . . . . . . . . . . 97 4.4.5 Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 97 4.5 Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 99 5 TWO-STAGE OPTIMIZER FRAMEWORK 5.1 Upper-Bound-Driven Join Ordering Component . . . . . . . . . . . . . 101 5.2 Physical Operator Selection Component . . . . . . . . . . . . . . . . . . 103 5.3 Example Query Optimization . . . . . . . . . . . . . . . . . . . . . . . . . 103 6 CONCLUSION 107 BIBLIOGRAPHY 109 LIST OF FIGURES 117 LIST OF TABLES 121 A APPENDIX A.1 Basics of Query Execution . . . . . . . . . . . . . . . . . . . . . . . . . . . 123 A.2 Why Q? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 124 A.3 0-TS Proof of Unbiased Estimate . . . . . . . . . . . . . . . . . . . . . . . . 125 A.4 UES Upper Bound Property . . . . . . . . . . . . . . . . . . . . . . . . . . . 127 A.5 TONIC – Selectivity-Aware Branching . . . . . . . . . . . . . . . . . . . . . 128 A.6 TONIC – Sequences of Query Execution . . . . . . . . . . . . . . . . . . . 129

Querying and efficiently analyzing complex data is required to gain valuable business insights, to support machine learning applications, and to make up-to-date information available. Therefore, this thesis investigates opportunities and challenges of selecting the most efficient execution strategy for analytical queries. These challenges include hard-to-capture data characteristics such as skew and correlation, the support of arbitrary data types, and the optimization time overhead of complex queries. Existing approaches often rely on optimistic assumptions about the data distribution, which can result in significant response time delays when these assumptions are not met. On the contrary, we focus on robust query optimization, emphasizing consistent query performance and applicability. Our presentation follows the general select-project-join query pattern, representing the fundamental stages of analytical query processing. To support arbitrary data types and complex filter expressions in the select stage, a novel sampling-based selectivity estimator is developed. Our approach exploits information from filter subexpressions and estimates correlations that are not captured by existing sampling-based methods. We demonstrate improved estimation accuracy and query execution time. Further, to minimize the runtime overhead of sampling, we propose new techniques that exploit access patterns and auxiliary database objects such as indices. For the join stage, we introduce a robust optimization approach by developing an upper-bound join enumeration strategy that connects accurate filter selectivity estimates –e.g., using our sampling-based approach– to join ordering. We demonstrate that join orders based on our upper-bound join ordering strategy achieve more consistent performance and faster workload execution on state-of-the-art database systems. However, besides identifying good logical join orders, it is crucial to determine appropriate physical join operators before query plan execution. To understand the importance of fine-grained physical operator selections, we exhaustively execute fixed join orders with all possible operator combinations. This analysis reveals that none of the investigated query optimizers fully reaches the potential of optimal operator decisions. Based on these insights and to achieve fine-grained operator selections for the previously determined join orders, the thesis presents a lightweight learning-based physical execution plan refinement component called. We show that this refinement component consistently outperforms existing approaches for physical operator selection while enabling a novel two-stage optimizer design. We conclude the thesis by providing a framework for the two-stage optimizer design that allows users to modify, replicate, and further analyze the concepts discussed throughout this thesis.:1 INTRODUCTION 1.1 Analytical Query Processing . . . . . . . . . . . . . . . . . . . 12 1.2 Select-Project-Join Queries . . . . . . . . . . . . . . . . . . . 13 1.3 Basics of SPJ Query Optimization . . . . . . . . . . . . . . . . . 14 1.3.1 Plan Enumeration . . . . . . . . . . . . . . . . . . . . . . . . 14 1.3.2 Cost Model . . . . . . . . . . . . . . . . . . . . . . . . . . . 15 1.3.3 Cardinality Estimation . . . . . . . . . . . . . . . . . . . . . 15 1.4 Robust SPJ Query Optimization . . . . . . . . . . . . . . . . . . 16 1.4.1 Tail Latency Root Cause Analysis . . . . . . . . . . . . . . . . 17 1.4.2 Tenets of Robust Query Optimization . . . . . . . . . . . . . . 19 1.5 Contribution . . . . . . . . . . . . . . . . . . . . . . . . . . . 19 1.6 Outline . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20 2 SELECT (-PROJECT) STAGE 2.1 Sampling for Selectivity Estimation . . . . . . . . . . . . . . . 24 2.2 Related Work . . . . . . . . . . . . . . . . . . . . . . . . . . . 28 2.2.1 Combined Selectivity Estimation (CSE) . . . . . . . . . . . . . 29 2.2.2 Kernel Density Estimator . . . . . . . . . . . . . . . . . . . . 31 2.2.3 Machine Learning . . . . . . . . . . . . . . . . . . . . . . . . 32 2.3 Beta Estimator for 0-Tuple-Situations . . . . . . . . . . . . . . 33 2.3.1 Methodology . . . . . . . . . . . . . . . . . . . . . . . . . . 33 2.3.2 Beta Distribution in Non-0-TS . . . . . . . . . . . . . . . . . 35 2.3.3 Parameter Estimation in 0-TS . . . . . . . . . . . . . . . . . . 37 2.3.4 Selectivity Estimation and Predicate Ordering . . . . . . . . . 39 2.3.5 Evaluation . . . . . . . . . . . . . . . . . . . . . . . . . . . 46 2.4 Customized Sampling Techniques . . . . . . . . . . . . . . . . . . 53 2.4.1 Focused Sampling . . . . . . . . . . . . . . . . . . . . . . . . 54 2.4.2 Conditional Sampling . . . . . . . . . . . . . . . . . . . . . . 56 2.4.3 Zone Pruning . . . . . . . . . . . . . . . . . . . . . . . . . . 58 2.4.4 Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . 59 2.5 Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59 3 JOIN STAGE: LOGICAL ENUMERATION 3.1 Related Work . . . . . . . . . . . . . . . . . . . . . . . . . . . 62 3.1.1 Point Estimates . . . . . . . . . . . . . . . . . . . . . . . . 63 3.1.2 Join Cardinality Upper Bound . . . . . . . . . . . . . . . . . . 64 3.2 Upper Bound Join Enumeration with Synopsis (UES) . . . . . . . . . 66 3.2.1 U-Block: Simple Upper Bound for Joins . . . . . . . . . . . . . 67 3.2.2 E-Block: Customized Enumeration Scheme . . . . . . . . . . . . . 68 3.2.3 UES Algorithm . . . . . . . . . . . . . . . . . . . . . . . . . 69 3.3 Evaluation . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71 3.3.1 General Performance . . . . . . . . . . . . . . . . . . . . . . 72 3.3.2 Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . 74 3.4 Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 76 4 JOIN STAGE: PHYSICAL OPERATOR SELECTION 4.1 Operator Selection vs Join Ordering . . . . . . . . . . . . . . . 77 4.2 Related Work . . . . . . . . . . . . . . . . . . . . . . . . . . . 80 4.2.1 Adaptive Query Processing . . . . . . . . . . . . . . . . . . . 80 4.2.2 Bandit Optimizer (Bao) . . . . . . . . . . . . . . . . . . . . . 81 4.3 TONIC: Learned Physical Join Operator Selection . . . . . . . . . 82 4.3.1 Query Execution Plan Synopsis (QEP-S) . . . . . . . . . . . . . 83 4.3.2 QEP-S Life-Cycle . . . . . . . . . . . . . . . . . . . . . . . . 84 4.3.3 QEP-S Design Considerations . . . . . . . . . . . . . . . . . . 87 4.4 Evaluation . . . . . . . . . . . . . . . . . . . . . . . . . . . . 89 4.4.1 Performance Factors . . . . . . . . . . . . . . . . . . . . . . 90 4.4.2 Rate of Improvement . . . . . . . . . . . . . . . . . . . . . . 92 4.4.3 Data Shift . . . . . . . . . . . . . . . . . . . . . . . . . . . 95 4.4.4 TONIC - Runtime Traits . . . . . . . . . . . . . . . . . . . . . 97 4.4.5 Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . 97 4.5 Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 99 5 TWO-STAGE OPTIMIZER FRAMEWORK 5.1 Upper-Bound-Driven Join Ordering Component . . . . . . . . . . . . 101 5.2 Physical Operator Selection Component . . . . . . . . . . . . . . 103 5.3 Example Query Optimization . . . . . . . . . . . . . . . . . . . . 103 6 CONCLUSION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 107 BIBLIOGRAPHY . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 109 LIST OF FIGURES . . . . . . . . . . . . . . . . . . . . . . . . . . . 117 LIST OF TABLES . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121 A APPENDIX A.1 Basics of Query Execution . . . . . . . . . . . . . . . . . . . . 123 A.2 Why Q? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 124 A.3 0-TS Proof of Unbiased Estimate . . . . . . . . . . . . . . . . . 125 A.4 UES Upper Bound Property . . . . . . . . . . . . . . . . . . . . . 127 A.5 TONIC – Selectivity-Aware Branching . . . . . . . . . . . . . . . 128 A.6 TONIC – Sequences of Query Execution . . . . . . . . . . . . . . . 129

<p>Modulation of neural activity with electrical stimulation is a widespread therapy for treating neurological disorders and diseases. Two notable applications that have had striking clinical success are deep brain stimulation (DBS) for the treatment of movement disorders (e.g., Parkinson's disease) and spinal cord stimulation (SCS) for the treatment of chronic low back and limb pain. In these therapies, the battery life of the stimulators is much less than the required duration of treatment, requiring patients to undergo repeated battery replacement surgeries, which are costly and obligate them to incur repeatedly the risks associated with surgery. Further, deviations in lead position of 2-3 mm can preclude some or all potential clinical benefits, and in some cases, generate side-effects by stimulation of non-target regions. Therefore, despite the success of DBS and SCS, their efficiency and ability to activate target neural elements over non-target elements, termed selectivity, are inadequate and need improvement.</p><p>We combined computational models of volume conduction in the brain and spine with cable models of neurons to design novel electrode configurations for efficient and selective electrical stimulation of nervous tissue. We measured the efficiency and selectivity of prototype electrode designs in vitro and in vivo. Stimulation efficiency was increased by increasing electrode area and/or perimeter, but the effect of increasing perimeter was not as pronounced as increasing area. Cylindrical electrodes with aspect (height to diameter) ratios of > 5 were the most efficient for stimulating neural elements oriented perpendicular to the axis of the electrode, whereas electrodes with aspect ratios of < 2 were the most efficient for stimulating parallel neural elements.</p><p>Stimulation selectivity was increased by combining two or more electrodes in multipolar configurations. Asymmetric bipolar configurations were optimal for activating parallel axons over perpendicular axons; arrays of cathodes with short interelectrode spacing were optimal for activating perpendicular axons over parallel axons; anodes displaced from the center of the target region were optimal for selectively activating terminating axons over passing axons; and symmetric tripolar configurations were optimal for activating neural elements based on their proximity to the electrode. The performance of the efficient and selective designs was not be explained solely by differences in their electrical properties, suggesting that field-shaping effects from changing electrode geometry and polarity can be as large as or larger than the effects of decreasing electrode impedance.</p><p>Advancing our understanding of the features of electrode geometry that are important for increasing stimulation efficiency and selectivity facilitates the design of the next generation of stimulation electrodes for the brain and spinal cord. Increased stimulation efficiency will increase the battery life of IPGs, increase the recharge interval of rechargeable IPGs, and potentially reduce stimulator volume. Greater selectivity may improve the success rate of DBS and SCS by mitigating the sensitivity of clinical outcomes to malpositioning of the electrode.</p> / Dissertation

<p>The neuromodulator dopamine signals through the dopamine D2 receptor (D2R) to modulate central nervous system functions through diverse signal transduction pathways. D2R is a prominent target for drug treatments in disorders where dopamine function is aberrant, such as schizophrenia. D2R signals through distinct G protein and β-arrestin pathways and drugs that are functionally selective for these pathways could have improved therapeutic potential. How D2R signals through the two pathways is still not well defined, and efforts to elucidate these pathways have been hampered by the lack of adequate tools for assessing the contribution of each pathway independently. To address this, Evolutionary Trace was used to produce D2R mutants with strongly biased interactions for either G protein or β-arrestin. Additionally, various permutations of these mutants were used to identify critical determinants of D2R functional selectivity. D2R interactions with the two major downstream signal transducers were effectively dissociated and G protein signaling accounts for D2R canonical MAP kinase signaling cascade activation. Nevertheless, when expressed in mice, the β-arrestin biased D2R caused a significant potentiation of amphetamine-induced locomotion, while the G protein biased D2R had minimal effects. The mutant receptors generated here provide a new molecular tool set that enable a better definition of the individual roles of G protein and β-arrestin signaling in D2R pharmacology, neurobiology and associated pathologies.</p> / Dissertation

<p>Evolution of enzymes with novel functional properties has gained much attention in recent years. Naturally evolved enzymes are adapted to work in living cells under physiological conditions, circumstances that are not always available for industrial processes calling for novel and better catalysts. Furthermore, altering enzyme function also affords insight into how enzymes work and how natural evolution operates. </p><p>Previous investigations have explored catalytic properties in the directed evolution of mutant libraries with high sequence variation. Before this study was initiated, functional analysis of mutant libraries was, to a large extent, restricted to uni- or bivariate methods. Consequently, there was a need to apply multivariate data analysis (MVA) techniques in this context. Directed evolution was approached by DNA shuffling of glutathione transferases (GSTs) in this thesis. GSTs are multifarious enzymes that have detoxication of both exo- and endogenous compounds as their primary function. They catalyze the nucleophilic attack by the tripeptide glutathione on many different electrophilic substrates. </p><p>Several multivariate analysis tools, <i>e.g.</i> principal component (PC), hierarchical cluster, and K-means cluster analyses, were applied to large mutant libraries assayed with a battery of GST substrates. By this approach, evolvable units (quasi-species) fit for further evolution were identified. It was clear that different substrates undergoing different kinds of chemical transformation can group together in a multi-dimensional substrate-activity space, thus being responsible for a certain quasi-species cluster. Furthermore, the importance of the chemical environment, or substrate matrix, in enzyme evolution was recognized. Diverging substrate selectivity profiles among homologous enzymes acting on substrates performing the same kind of chemistry were identified by MVA. Important structure-function activity relationships with the prodrug azathioprine were elucidated by segment analysis of a shuffled GST mutant library. Together, these results illustrate important methods applied to molecular enzyme evolution.</p>