Development of the Ultrashort Pulse Nonlinear Optical Microscopy Spectral Imaging System

Nonlinear Optical Microscopy (NLOM) has been shown to be a valuable tool for noninvasive imaging of complex biological systems. An effective approach for multicolor molecular microscopy is simultaneous excitation of multiple fluorophores by broadband sub-10-fs pulses. This dissertation will discuss the development of two spectral imaging systems using the principles of nonlinear optical microscopy for pixel-by-pixel spectral segmentation of multiple fluorescent spectra. The first spectral system is reliant on a fiber-optic cable to transmit fluorescent signal to a spectrometer, while the second is based on a spectrometer with an aberration-corrected concave grating that is directly coupled to the microscope. A photon-counting, 16-channel multianode photomultiplier tube (PMT) is used for both systems. Custom software developed in LabVIEW controls multiple counter cards as well as a field-programmable gate array (FPGA) for 1 Hz acquisition of 256x256x16 spectral images. Biological specimen consisting of multicolor endothelial cells and zebrafish will be used for experimental verification. Results indicate successful spectral segmentation of multiple fluorophores with a decrease in signal-to-noise ratio in the FPGA-based imaging system

Production of FMDV virus-like particles by a SUMO fusion protein approach in Escherichia coli

Virus-like particles (VLPs) are formed by the self-assembly of envelope and/or capsid proteins from many viruses. Some VLPs have been proven successful as vaccines, and others have recently found applications as carriers for foreign antigens or as scaffolds in nanoparticle biotechnology. However, production of VLP was usually impeded due to low water-solubility of recombinant virus capsid proteins. Previous studies revealed that virus capsid and envelope proteins were often posttranslationally modified by SUMO in vivo, leading into a hypothesis that SUMO modification might be a common mechanism for virus proteins to retain water-solubility or prevent improper self-aggregation before virus assembly. We then propose a simple approach to produce VLPs of viruses, e.g., foot-and-mouth disease virus (FMDV). An improved SUMO fusion protein system we developed recently was applied to the simultaneous expression of three capsid proteins of FMDV in E. coli. The three SUMO fusion proteins formed a stable heterotrimeric complex. Proteolytic removal of SUMO moieties from the ternary complexes resulted in VLPs with size and shape resembling the authentic FMDV. The method described here can also apply to produce capsid/envelope protein complexes or VLPs of other disease-causing viruses

Charged Dilatonic Black Holes and their Transport Properties

We briefly explain the consistency conditions imposed on the effective holographic theories, which are parameterized by two real exponents $(\gamma,\delta)$ that control the IR dynamics. The general scaling of DC resistivity with temperature at low temperature and AC conductivity at low frequency across the whole $(\gamma,\delta)$ plane are explained. There is a codimension-one region where the DC resistivity is linear in the temperature. For massive carriers, it is shown that when the scalar operator is not the dilaton, the DC resistivity scales as the heat capacity (and entropy) for $(2+1)$-dimensional systems. Regions are identified where the theory at finite density is a Mott-like insulator. This contribution is based on arXiv:1005.4690 with emphasis on the transport properties of charged dilatonic black holes with potential.Comment: 6 pages and 6 figures. Contribution to the proceedings of the XVIth European Workshop on String Theory, Madrid, Spain, June 14-18 201

Unscented Kalman Filtering for Single Camera Based Motion and Shape Estimation

Accurate estimation of the motion and shape of a moving object is a challenging task due to great variety of noises present from sources such as electronic components and the influence of the external environment, etc. To alleviate the noise, the filtering/estimation approach can be used to reduce it in streaming video to obtain better estimation accuracy in feature points on the moving objects. To deal with the filtering problem in the appropriate nonlinear system, the extended Kalman filter (EKF), which neglects higher-order derivatives in the linearization process, has been very popular. The unscented Kalman filter (UKF), which uses a deterministic sampling approach to capture the mean and covariance estimates with a minimal set of sample points, is able to achieve at least the second order accuracy without Jacobians’ computation involved. In this paper, the UKF is applied to the rigid body motion and shape dynamics to estimate feature points on moving objects. The performance evaluation is carried out through the numerical study. The results show that UKF demonstrates substantial improvement in accuracy estimation for implementing the estimation of motion and planar surface parameters of a single camera

Increased Risk for Entamoeba histolytica Infection and Invasive Amebiasis in HIV Seropositive Men Who Have Sex with Men in Taiwan

Entamoeba histolytica, morphologically identical to but genetically different from E. dispar and E. moshkovskii, is the causative agent of amebiasis. Recently there have been reports of increased risk for amebiasis among men who have sex with men (MSM) due to oral-anal sexual contact in several developed countries. In this longitudinal follow-up study, the incidence of amebiasis was determined among HIV-infected patients using serological and specific amebic antigen assays. DNA extracted from stool samples containing E. histolytica were analyzed by PCR, sequenced, and compared. Clinical manifestations and treatment response of invasive amebiasis in HIV-infected patients were reviewed. The results demonstrated that HIV-infected MSM were at significantly higher risk of amebiasis than patients from other risk groups. Clustering of E. histolytica isolates by sequencing analyses from geographically unrelated patients suggested person-to-person transmission. Despite immunosuppression, amebic liver abscesses and colitis responded favorably to metronidazole therapy. It is important to investigate in areas of high incidence of both amebiasis and HIV (sub-Saharan Africa) how generalizable these findings are

Measuring anisotropic scattering in the cuprates

A simple model of anisotropic scattering in a quasi two-dimensional metal is studied. Its simplicity allows an analytic calculation of transport properties using the Boltzmann equation and relaxation time approximation. We argue that the c-axis magnetoresistance provides the key test of this model of transport. We compare this model with experiments on overdoped Tl-2201 and find reasonable agreement using only weak scattering anisotropy. We argue that optimally doped Tl-2201 should show strong angular-dependent magnetoresistance within this model and would provide a robust way of determining the in-plane scattering anisotropy in the cuprates.Comment: 12 pages, 8 figures, typset in REVTeX 4. Version 2; added references and corrected typo

Low-dose versus standard-dose alteplase in acute ischemic stroke in Asian stroke registries: an individual patient data pooling study

10.1177/1747493019858777INTERNATIONAL JOURNAL OF STROKE147670-67

Three New Structures of Left-Handed RadA Helical Filaments: Structural Flexibility of N-Terminal Domain Is Critical for Recombinase Activity

RecA family proteins, including bacterial RecA, archaeal RadA, and eukaryotic Dmc1 and Rad51, mediate homologous recombination, a reaction essential for maintaining genome integrity. In the presence of ATP, these proteins bind a single-strand DNA to form a right-handed nucleoprotein filament, which catalyzes pairing and strand exchange with a homologous double-stranded DNA (dsDNA), by as-yet unknown mechanisms. We recently reported a structure of RadA left-handed helical filament, and here present three new structures of RadA left-handed helical filaments. Comparative structural analysis between different RadA/Rad51 helical filaments reveals that the N-terminal domain (NTD) of RadA/Rad51, implicated in dsDNA binding, is highly flexible. We identify a hinge region between NTD and polymerization motif as responsible for rigid body movement of NTD. Mutant analysis further confirms that structural flexibility of NTD is essential for RadA's recombinase activity. These results support our previous hypothesis that ATP-dependent axial rotation of RadA nucleoprotein helical filament promotes homologous recombination