Prior image-constrained l(1)-norm-based reconstruction method for effective usage of structural information in diffuse optical tomography

A novel approach that can more effectively use the structural information provided by the traditional imaging modalities in multimodal diffuse optical tomographic imaging is introduced. This approach is based on a prior image-constrained-l(1) minimization scheme and has been motivated by the recent progress in the sparse image reconstruction techniques. It is shown that the proposed framework is more effective in terms of localizing the tumor region and recovering the optical property values both in numerical and gelatin phantom cases compared to the traditional methods that use structural information. (C) 2012 Optical Society of America

Warped alternatives to Froggatt-Nielsen models

We consider the Randall-Sundrum (RS) setup to be a theory of flavor, as an alternative to Froggatt-Nielsen models instead of as a solution to the hierarchy problem. The RS framework is modified by taking the low-energy brane to be at the grand unified theory (GUT) scale. This also alleviates constraints from flavor physics. Fermion masses and mixing angles are fit at the GUT scale. The ranges of the bulk mass parameters are determined using a chi(2) fit taking into consideration the variation in O(1) parameters. In the hadronic sector, the heavy top quark requires large bulk mass parameters localizing the right-handed top quark close to the IR brane. Two cases of neutrino masses are considered: (a) Planck scale lepton number violation and (b) Dirac neutrino masses. Contrary to the case of weak scale RS models, both these cases give reasonable fits to the data, with the Planck scale lepton number violation fitting slightly better compared to the Dirac case. In the supersymmetric version, the fits are not significantly different except for the variation in tan beta. If the Higgs superfields and the supersymmetry breaking spurion are localized on the same brane, then the structure of the sfermion masses are determined by the profiles of the zero modes of the hypermultiplets in the bulk. Trilinear terms have the same structure as the Yukawa matrices. The resultant squark spectrum is around similar to 2-3 TeV required by the light Higgs mass to be around 125 GeV and to satisfy the flavor violating constraints.

Simultaneous source localization and boundary mapping for contaminants

This paper addresses the problem of localizing the sources of contaminants spread in the environment, and mapping the boundary of the affected region using an innovative swarm intelligence based technique. Unlike most work in this area the algorithm is capable of localizing multiple sources simultaneously while also mapping the boundary of the contaminant spread. At the same time the algorithm is suitable for implementation using a mobile robotic sensor network. Two types of agents, called the source localization agents (or S-agents) and boundary mapping agents (or B-agents) are used for this purpose. The paper uses the basic glowworm swarm optimization (GSO) algorithm, which has been used only for multiple signal source localization, and modifies it considerably to make it suitable for both these tasks. This requires the definition of new behaviour patterns for the agents based on their terminal performance as well as interactions between them that helps the swarm to split into subgroups easily and identify contaminant sources as well as spread along the boundary to map its full length. Simulations results are given to demonstrate the efficacy of the algorithm.

Ferrocenyl-L-amino acid copper(II) complexes showing remarkable photo-induced anticancer activity in visible light

Ferrocene-conjugated copper(II) complexes Cu(Fc-aa)(aip)](ClO4) (1-3) and (Cu(Fc-aa)(pyip)](ClO4) (4-6) of L-amino acid reduced Schiff bases (Fc-aa), 2-(9-anthryl)-1H-imidazo4,5-f]1,10]phenanthroline (aip) and 2-(1-pyrenyl)-1H-imidazo4,5-f] 1,10]phenanthroline (pyip), where Fc-aa is ferrocenylmethyl-L-tyrosine (Fc-Tyr in 1, 4), ferrocenylmethyl-L-tryptophan (Fc-Trp in 2, 5) and ferrocenylmethyl-L-methionine (Fc-Met in 3, 6), were prepared and characterized, and their photocytotoxicity was studied (Fc = ferrocenyl moiety). Phenyl analogues, viz. (Cu(Ph-Met)(aip)](ClO4) (7) and (Cu(Ph-Met)(pyip)](ClO4) (8), were prepared and used as control compounds. The bis-imidazophenanthroline copper(II) complexes, viz. (Cu(aip)(2)(NO3)](NO3) (9) and Cu(pyip)(2)(NO3)](NO3) (10), were also prepared and used as controls. Complexes 1-6 having a redox inactive cooper(II) center showed the Fc(+)-Fc redox couple at similar to 0.5 V vs. SCE in DMF-0.1 mol (Bu4N)-N-n](ClO4). The copper(II)-based d-d band was observed near 600 nm in DMF-Tris-HCl buffer (1 :1 v/v). The ferrocenyl complexes showed low dark toxicity, but remarkably high photocytotoxicity in human cervical HeLa and human breast adenocarcinoma MCF-7 cancer cells giving an excellent photo-dynamic effect while their phenyl analogues were inactive. The photo-exposure caused significant morphological changes in the cancer cells when compared to the non-irradiated ones. The photophysical processes were rationalized from the theoretical studies. Fluorescence microscopic images showed 3 and 6 localizing predominantly in the endoplasmic reticulum (ER) of the cancer cells, thus minimizing any undesirable effects involving nuclear DNA.

Tailoring the interface of an immiscible polymer blend by a mutually miscible homopolymer grafted onto graphene oxide: outstanding mechanical properties

A unique strategy was adopted here to improve the compatibility between the components of an immiscible polymer blend and strengthen the interface. PMMA, a mutually miscible polymer to both PVDF and ABS, improved the compatibility between the phases by localizing at the blends interface. This was supported by the core-shell formation with PMMA as the shell and ABS as the core as observed from the SEM micrographs. This phenomenon was strongly contingent on the concentration of PMMA in the blends. This strategy was further extended to localize graphene oxide (GO) sheets at the blends interface by chemically coupling it to PMMA (PMMA-g-GO). A dramatic increment of ca. 84% in the Young's modulus and ca. 124% in the yield strength was observed in the presence of PMMA-g-GO with respect to the neat blends. A simultaneous increment in both the strength and the modulus was observed in the presence of PMMA-g-GO whereas, only addition of GO resulted in a moderate improvement in the yield strength. This study reveals that a mutually miscible polymer can render compatibility between the immiscible pair and can improve the stress transfer at the interface.

Tailoring the dispersion of multiwall carbon nanotubes in co-continuous PVDF/ABS blends to design materials with enhanced electromagnetic interference shielding

Highly conducting composites were derived by selectively localizing multiwall carbon nanotubes (MWNTs) in co-continuous PVDF/ABS (50/50, wt/wt) blends. The electrical percolation threshold was obtained between 0.5 and 1 wt% MWNTs as manifested by a dramatic increase in the electrical conductivity by about six orders of magnitude with respect to the neat blends. In order to further enhance the electrical conductivity of the blends, the MWNTs were modified with amine terminated ionic liquid (IL), which, besides enhancing the interfacial interaction with PVDF, facilitated the formation of a network like structure of MWNTs. This high electrical conductivity of the blends, at a relatively low fraction (1 wt%), was further explored to design materials that can attenuate electromagnetic (EM) radiation. More specifically, to attenuate the EM radiation by absorption, a ferroelectric phase was introduced. To accomplish this, barium titanate (BT) nanoparticles chemically stitched onto graphene oxide (GO) sheets were synthesized and mixed along with MWNTs in the blends. Intriguingly, the total EM shielding effectiveness (SE) was enhanced by ca. 10 dB with respect to the blends with only MWNTs. In addition, the effect of introducing a ferromagnetic phase (Fe3O4) along with IL modified MWNTs was also investigated. This study opens new avenues in designing materials that can attenuate EM radiation by selecting either a ferroelectric (BT-GO) or a ferromagnetic phase (Fe3O4) along with intrinsically conducting nanoparticles (MWNTs).

Microwave absorbers designed from PVDF/SAN blends containing multiwall carbon nanotubes anchored cobalt ferrite via a pyrene derivative

Lightweight and flexible electromagnetic shielding materials were designed by selectively localizing multiwall carbon nanotubes (MWNTs) anchored magnetic nanoparticles in melt mixed co-continuous blends of polyvinylidene fluoride (PVDF) and poly(styrene-co-acrylonitrile) (SAN). In order to facilitate better dispersion, the MWNTs were modified using pyrenebutyric acid (PBA) via pi-pi stacking. While one of the two-targeted properties, i.e., high electrical conductivity, was achieved by PBA modified MWNTs, high magnetic loss was accomplished by introducing nickel (NF) or cobalt ferrites (CF). Moreover, the attenuation by absorption can be tuned either by using NF (58% absorption) or CF (64% absorption) in combination with PBA-MWNTs. More interestingly, when CF was anchored on to MWNTs via the pyrene derivative, the minimum reflection loss attained was -55 dB in the Ku band (12-18 GHz) frequency and with a large bandwidth. In addition, the EM waves were blocked mostly by absorption (70%). This study opens new avenues in designing flexible and lightweight microwave absorbers.

Engineering Nanostructures by Decorating Magnetic Nanoparticles onto Graphene Oxide Sheets to Shield Electromagnetic Radiations

In this study, a minimum, reflection loss of 70 a was achieved, for a 6 mm thick shield (at 17.1 GHz frequency) employing a unique approach. This was accomplished by engineering nanostructures through decoration of magnetic nanopartides (nickel, Ni) onto graphene oxide (GO) sheets. Enhanced electromagnetic (EM) shielding was derived by selectively, localizing the nanoscopic particles in a specific phase of polyethylene (PE)/poly(ethylene oxide) (PEO) blends. By introduction of a conducting inclusion (like multiwall carbon nanotubes, MWNTs) together with the engineered nanostructures (nickel-decorated GO, (GO-Ni), the shielding efficiency can be enhanced significantly in contrast to physically mixing the particles in the blends. For instance, the composites showed a shielding efficiency >25 dB for a combination of MWNTS (3 wt %) and Ni nanoparticles (52 wt %) in PE/PEO blends. However, similar shielding effectiveness could be achieved for a combination of MWNTs (3 wt %) and 10 vol % of GO-Ni where in the effective concentration of Ni was only 19 wt %. The GO-Ni sheets facilitated in an efficient charge transfer as manifested from high electrical conductivity in the blends besides enhancing the permeability in the blends. It is envisioned that GO is simultaneously reduced in the process of synthesizing GO-Ni, and this facilitated in efficient charge transfer between the neighboring CNTs. More interestingly, the blends With MWNTs/GO-Ni attenuated the incoming EM radiation mostly by absorption. This study opens new avenues in designing polyolefin-based lightweight shielding materials by engineering nanostructures for numerous applications.

Attenuating microwave radiation by absorption through controlled nanoparticle localization in PC/PVDF blends

Nanoscale ordering in a polymer blend structure is indispensable to obtain materials with tailored properties. It was established here that controlling the arrangement of nanoparticles, with different characteristics, in co-continuous PC/PVDF (polycarbonate/poly(vinylidene fluoride)) blends can result in outstanding microwave absorption (ca. 90%). An excellent reflection loss (RL) of ca. -71 dB was obtained for a model blend structure wherein the conducting (multiwall carbon nanotubes, MWNTs) and the magnetic inclusions (Fe3O4) are localized in PVDF and the dielectric inclusion (barium titanate, BT) is in PC. The MWNTs were modified using polyaniline, which facilitates better charge transport in the blends. Furthermore, by introducing surface active groups on BT nanoparticles and changing the macroscopic processing conditions, the localization of BT nanoparticles can be tailored, otherwise BT nanoparticles would localize in the preferred phase (PVDF). In this study, we have shown that by ordered arrangement of nanoparticles, the incoming EM radiation can be attenuated. For instance, when PANI-MWNTs were localized in PVDF, the shielding was mainly through reflection. Now by localizing the conducting inclusion and the magnetic lossy materials in PVDF and the dielectric materials in PC, an outstanding shielding effectiveness of ca. -37 dB was achieved where shielding was mainly through absorption (ca. 90%). Thus, this study clearly demonstrates that lightweight microwave absorbers can be designed using polymer blends as a tool.

Tailor-Made Distribution of Nanoparticles in Blend Structure toward Outstanding Electromagnetic Interference Shielding

Engineering blend structure with tailor-made distribution of nanoparticles is the prime requisite to obtain materials with extraordinary properties. Herein, a unique strategy of distributing nanoparticles in different phases of a blend structure has resulted in >99% blocking of incoming electromagnetic (EM) radiation. This is accomplished by designing a ternary polymer blend structure using polycarbonate (PC), poly(vinylidene fluoride) (PVDF), and poly(methyl methacrylate) (PMMA) to simultaneously improve the structural, electrical, and electromagnetic interference shielding (EMI). The blend structure was made conducting by preferentially localizing the multi-wall nanotubes (MWNTs) in the PVDF phase. By taking advantage of pp stacking MWNTs was noncovalently modified with an imidazolium based ionic liquid (IL). Interestingly, the enhanced dispersion of IL-MWNTs in PVDF improved the electrical conductivity of the blends significantly. While one key requisite to attenuate EM radiation (i.e., electrical conductivity) was achieved using MWNTs, the magnetic properties of the blend structure was tuned by introducing barium ferrite (BaFe) nanoparticles, which can interact with the incoming EM radiation. By suitably modifying the surface of BaFe nanoparticles, we can tailor their localization under the macroscopic processing condition. The precise localization of BaFe nanoparticles in the PC phase, due to nucleophilic substitution reaction, and the MWNTs in the PVDF phase not only improved the conductivity but also facilitated in absorption of the incoming microwave radiation due to synergetic effect from MWNT and BaFe. The shielding effectiveness (SE) was measured in X and K-u band, and an enhanced SE of -37 dB was noted at 18 GHz frequency. PMMA, which acted as an interfacial modifier in PC/PVDF blends further, resulting in a significant enhancement in the mechanical properties besides retaining high SE. This study opens a new avenue in designing mechanically strong microwave absorbers with a suitable combination of materials.

U.S. Caribbean fish trap fishery costs and earnings study

Executive Summary: This study describes the socio-economic characteristics of the U.S. Caribbean trap fishery that encompasses the Commonwealth of Puerto Rico and Territory of the U.S. Virgin Islands. In-person interviews were administered to one hundred randomly selected trap fishermen, constituting nearly 25% of the estimated population. The sample was stratified by geographic area and trap tier. The number of traps owned or fished to qualify for a given tier varied by island. In Puerto Rico, tier I consisted of fishermen who had between 1-40 fish traps, tier II was made up of fishermen who possessed between 41 and 100 fish traps, and tier III consisted of fishermen who held in excess of 100 fish traps. In St. Thomas and St. John, tier I was composed of fishermen who held between 1 and 50 fish traps, tier II consisted of fishermen who had between 51-150 fish traps and tier III was made up of fishermen who had in excess of 150 fish traps. Lastly, in St. Croix, tier I was made up of fishermen who had less than 20 fish traps and tier II consisted of fishermen who had 20 or more fish traps. The survey elicited information on household demographics, annual catch and revenue, trap usage, capital investment on vessels and equipment, fixed and variable costs, behavioral response to a hypothetical trap reduction program and the spatial distribution of traps. The study found that 79% of the sampled population was 40 years or older. The typical Crucian trap fisherman was older than their Puerto Rican and St. Thomian and St. Johnian counterparts. Crucian fishermen’s average age was 57 years whereas Puerto Rican fishermen’s average age was 51 years, and St. Thomian and St. Johnian fishermen’s average age was 48 years. As a group, St. Thomian and St. Johnian fishermen had 25 years of fishing experience, and Puerto Rican and Crucian fishermen had 30, and 29 years, respectively. Overall, 90% of the households had at least one dependent. The average number of dependents across islands was even, ranging between 2.8 in the district of St. Thomas and St. John and 3.4 in the district of St. Croix. The percentage utilization of catch for personal or family use was relatively low. Regionally, percentage use of catch for personal or family uses ranged from 2.5% in St. Croix to 3.8% in the St. Thomas and St. John. About 47% of the respondents had a high school degree. The majority of the respondents were highly dependent on commercial fishing for their household income. In St. Croix, commercial fishing made up 83% of the fishermen’s total household income, whereas in St. Thomas and St. John and Puerto Rico it contributed 74% and 68%, respectively. The contribution of fish traps to commercial fishing income ranged from 51% in the lowest trap tier in St. Thomas and St. John to 99% in the highest trap tier in St. Croix. On an island basis, the contribution of fish traps to fishing income was 75% in St. Croix, 61% in St. Thomas and St. John, and 59% in Puerto Rico. The value of fully rigged vessels ranged from $400 to $250,000. Over half of the fleet was worth $10,000 or less. The St. Thomas and St. John fleet reported the highest mean value, averaging $58,518. The Crucian and Puerto Rican fleets were considerably less valuable, averaging $19,831 and $8,652, respectively. The length of the vessels ranged from 14 to 40 feet. Fifty-nine percent of the sampled vessels were at least 23 feet in length. The average length of the St. Thomas and St. John fleet was 28 feet, whereas the fleets based in St. Croix and Puerto Rico averaged 21 feet. The engine’s propulsion ranged from 8 to 400 horsepower (hp). The mean engine power was 208 hp in St. Thomas and St. John, 108 hp in St. Croix, and 77 hp in Puerto Rico. Mechanical trap haulers and depth recorders were the most commonly used on-board equipment. About 55% of the sampled population reported owning mechanical trap haulers. In St. Thomas and St. John, 100% of the respondents had trap haulers compared to 52% in Puerto Rico and 20% in St. Croix. Forty-seven percent of the fishermen surveyed stated having depth recorders. Depth recorders were most common in the St. Thomas and St. John fleet (80%) and least common in the Puerto Rican fleet (37%). The limited presence of emergency position indication radio beacons (EPIRBS) and radar was the norm among the fish trap fleet. Only 8% of the respondents had EPIRBS and only 1% had radar. Interviewees stated that they fished between 1 and 350 fish traps. Puerto Rican respondents fished on average 39 fish traps, in contrast to St. Thomian and St. Johnian and Crucian respondents, who fished 94 and 27 fish traps, respectively. On average, Puerto Rican respondents fished 11 lobster traps, and St. Thomian and St. Johnian respondents fished 46 lobster traps. None of the Crucian respondents fished lobster traps. The number of fish traps built or purchased ranged between 0 and 175, and the number of lobster traps built or bought ranged between 0 and 200. Puerto Rican fishermen on average built or purchased 30 fish traps and 14 lobster traps, and St. Thomian and St. Johnian fishermen built or bought 30 fish traps and 11 lobster traps. Crucian fishermen built or bought 25 fish traps and no lobster traps. As a group, fish trap average life ranged between 1.3 and 5 years, and lobster traps lasted slightly longer, between 1.5 and 6 years. The study found that the chevron or arrowhead style was the most common trap design. Puerto Rican fishermen owned an average of 20 arrowhead traps. St. Thomian and St. Johnian and Crucian fishermen owned an average of 44 and 15 arrowhead fish traps, respectively. The second most popular trap design was the square trap style. Puerto Rican fishermen had an average of 9 square traps, whereas St. Thomian and St. Johnian fishermen had 33 traps and Crucian fishermen had 2 traps. Antillean Z (or S) -traps, rectangular and star traps were also used. Although Z (or S) -traps are considered the most productive trap design, fishermen prefer the smaller-sized arrowhead and square traps because they are easier and less expensive to build, and larger numbers of them can be safely deployed. The cost of a fish trap, complete with rope and buoys, varied significantly due to the wide range of construction materials utilized. On average, arrowhead traps commanded $94 in Puerto Rico, $251 in St. Thomas and St. John, and $119 in St. Croix. The number of trips per week ranged between 1 and 6. However, 72% of the respondents mentioned that they took two trips per week. On average, Puerto Rican fishermen took 2.1 trips per week, St. Thomian and St. Johnian fishermen took 1.4 trips per week, and Crucian fishermen took 2.5 trips per week. Most fishing trips started at dawn and finished early in the afternoon. Over 82% of the trips lasted 8 hours or less. On average, Puerto Rican fishermen hauled 27 fish traps per trip whereas St. Thomian and St. Johnian fishermen and Crucian fishermen hauled 68 and 26 fish traps per trip, respectively. The number of traps per string and soak time varied considerably across islands. In St. Croix, 84% of the respondents had a single trap per line, whereas in St. Thomas and St. John only 10% of the respondents had a single trap per line. Approximately, 43% of Puerto Rican fishermen used a single trap line. St. Thomian and St. Johnian fishermen soaked their traps for 6.9 days while Puerto Rican and Crucian fishermen soaked their traps for 5.7 and 3.6 days, respectively. The heterogeneity of the industry was also evidenced by the various economic surpluses generated. The survey illustrated that higher gross revenues did not necessarily translate into higher net revenues. Our analysis also showed that, on average, vessels in the trap fishery were able to cover their cash outlays, resulting in positive vessel income (i.e., financial profits). In Puerto Rico, annual financial profits ranged from $4,760 in the lowest trap tier to $32,467 in the highest tier, whereas in St. Thomas and St. John annual financial profits ranged from $3,744 in the lowest tier to $13,652 in the highest tier. In St. Croix, annual financial profits ranged between $9,229 and $15,781. The survey also showed that economic profits varied significantly across tiers. Economic profits measure residual income after deducting the remuneration required to keep the various factors of production in their existing employment. In Puerto Rico, annual economic profits ranged from ($9,339) in the lowest trap tier to $ 8,711 in the highest trap tier. In St. Thomas and St. John, annual economic profits ranged from ($7,920) in the highest tier to ($18,486) in the second highest tier. In St. Croix, annual economic profits ranged between ($7,453) to $10,674. The presence of positive financial profits and negative economic profits suggests that higher economic returns could be earned from a societal perspective by redirecting some of these scarce capital and human resources elsewhere in the economy. Furthermore, the presence of negative economic earnings is evidence that the fishery is overcapitalized and that steps need to be taken to ensure the long-run economic viability of the industry. The presence of positive financial returns provides managers with a window of opportunity to adopt policies that will strengthen the biological and economic performance of the fishery while minimizing any adverse impacts on local fishing communities. Finally, the document concludes by detailing how the costs and earnings information could be used to develop economic models that evaluate management proposals. (PDF contains 147 pages)

Subdiffusion of nonlinear waves in quasiperiodic potentials

19 p.

Estimation and inference for grasping and manipulation tasks using vision and kinesthetic sensors

This thesis presents a novel framework for state estimation in the context of robotic grasping and manipulation. The overall estimation approach is based on fusing various visual cues for manipulator tracking, namely appearance and feature-based, shape-based, and silhouette-based visual cues. Similarly, a framework is developed to fuse the above visual cues, but also kinesthetic cues such as force-torque and tactile measurements, for in-hand object pose estimation. The cues are extracted from multiple sensor modalities and are fused in a variety of Kalman filters.

A hybrid estimator is developed to estimate both a continuous state (robot and object states) and discrete states, called contact modes, which specify how each finger contacts a particular object surface. A static multiple model estimator is used to compute and maintain this mode probability. The thesis also develops an estimation framework for estimating model parameters associated with object grasping. Dual and joint state-parameter estimation is explored for parameter estimation of a grasped object's mass and center of mass. Experimental results demonstrate simultaneous object localization and center of mass estimation.

Dual-arm estimation is developed for two arm robotic manipulation tasks. Two types of filters are explored; the first is an augmented filter that contains both arms in the state vector while the second runs two filters in parallel, one for each arm. These two frameworks and their performance is compared in a dual-arm task of removing a wheel from a hub.

This thesis also presents a new method for action selection involving touch. This next best touch method selects an available action for interacting with an object that will gain the most information. The algorithm employs information theory to compute an information gain metric that is based on a probabilistic belief suitable for the task. An estimation framework is used to maintain this belief over time. Kinesthetic measurements such as contact and tactile measurements are used to update the state belief after every interactive action. Simulation and experimental results are demonstrated using next best touch for object localization, specifically a door handle on a door. The next best touch theory is extended for model parameter determination. Since many objects within a particular object category share the same rough shape, principle component analysis may be used to parametrize the object mesh models. These parameters can be estimated using the action selection technique that selects the touching action which best both localizes and estimates these parameters. Simulation results are then presented involving localizing and determining a parameter of a screwdriver.

Lastly, the next best touch theory is further extended to model classes. Instead of estimating parameters, object class determination is incorporated into the information gain metric calculation. The best touching action is selected in order to best discern between the possible model classes. Simulation results are presented to validate the theory.

Deep tissue fluorescence imaging with time-reversed light

Advances in optical techniques have enabled many breakthroughs in biology and medicine. However, light scattering by biological tissues remains a great obstacle, restricting the use of optical methods to thin ex vivo sections or superficial layers in vivo. In this thesis, we present two related methods that overcome the optical depth limit—digital time reversal of ultrasound encoded light (digital TRUE) and time reversal of variance-encoded light (TROVE). These two techniques share the same principle of using acousto-optic beacons for time reversal optical focusing within highly scattering media, like biological tissues. Ultrasound, unlike light, is not significantly scattered in soft biological tissues, allowing for ultrasound focusing. In addition, a fraction of the scattered optical wavefront that passes through the ultrasound focus gets frequency-shifted via the acousto-optic effect, essentially creating a virtual source of frequency-shifted light within the tissue. The scattered ultrasound-tagged wavefront can be selectively measured outside the tissue and time-reversed to converge at the location of the ultrasound focus, enabling optical focusing within deep tissues. In digital TRUE, we time reverse ultrasound-tagged light with an optoelectronic time reversal device (the digital optical phase conjugate mirror, DOPC). The use of the DOPC enables high optical gain, allowing for high intensity optical focusing and focal fluorescence imaging in thick tissues at a lateral resolution of 36 µm by 52 µm. The resolution of the TRUE approach is fundamentally limited to that of the wavelength of ultrasound. The ultrasound focus (~ tens of microns wide) usually contains hundreds to thousands of optical modes, such that the scattered wavefront measured is a linear combination of the contributions of all these optical modes. In TROVE, we make use of our ability to digitally record, analyze and manipulate the scattered wavefront to demix the contributions of these spatial modes using variance encoding. In essence, we encode each spatial mode inside the scattering sample with a unique variance, allowing us to computationally derive the time reversal wavefront that corresponds to a single optical mode. In doing so, we uncouple the system resolution from the size of the ultrasound focus, demonstrating optical focusing and imaging between highly diffusing samples at an unprecedented, speckle-scale lateral resolution of ~ 5 µm. Our methods open up the possibility of fully exploiting the prowess and versatility of biomedical optics in deep tissues.

Proteomic analysis of the Cdc48/Ubx network identifies a role for Ubx2 in the regulation of lipid biosynthesis

Cdc48/p97 is an essential, highly abundant hexameric member of the AAA (ATPase associated with various cellular activities) family. It has been linked to a variety of processes throughout the cell but it is best known for its role in the ubiquitin proteasome pathway. In this system it is believed that Cdc48 behaves as a segregase, transducing the chemical energy of ATP hydrolysis into mechanical force to separate ubiquitin-conjugated proteins from their tightly-bound partners.

Current models posit that Cdc48 is linked to its substrates through a variety of adaptor proteins, including a family of seven proteins (13 in humans) that contain a Cdc48-binding UBX domain. As such, due to the complexity of the network of adaptor proteins for which it serves as the hub, Cdc48/p97 has the potential to exert a profound influence on the ubiquitin proteasome pathway. However, the number of known substrates of Cdc48/p97 remains relatively small, and smaller still is the number of substrates that have been linked to a specific UBX domain protein. As such, the goal of this dissertation research has been to discover new substrates and better understand the functions of the Cdc48 network. With this objective in mind, we established a proteomic screen to assemble a catalog of candidate substrate/targets of the Ubx adaptor system.

Here we describe the implementation and optimization of a cutting-edge quantitative mass spectrometry method to measure relative changes in the Saccharomyces cerevisiae proteome. Utilizing this technology, and in order to better understand the breadth of function of Cdc48 and its adaptors, we then performed a global screen to identify accumulating ubiquitin conjugates in cdc48-3 and ubxΔ mutants. In this screen different ubx mutants exhibited reproducible patterns of conjugate accumulation that differed greatly from each other, pointing to various unexpected functional specializations of the individual Ubx proteins.

As validation of our mass spectrometry findings, we then examined in detail the endoplasmic-reticulum bound transcription factor Spt23, which we identified as a putative Ubx2 substrate. In these studies ubx2Δ cells were deficient in processing of Spt23 to its active p90 form, and in localizing p90 to the nucleus. Additionally, consistent with reduced processing of Spt23, ubx2Δ cells demonstrated a defect in expression of their target gene OLE1, a fatty acid desaturase. Overall, this work demonstrates the power of proteomics as a tool to identify new targets of various pathways and reveals Ubx2 as a key regulator lipid membrane biosynthesis.