Lyme disease caused by the Borrelia burgdorferi (Bb or B. burgdorferi) is the most common vector-borne, multi-systemic disease in the USA. Although most Lyme disease patients can be cured with a course of the first line of antibiotic treatment, some patients are intolerant to currently available antibiotics, necessitating the development of more effective therapeutics. We previously found several drugs, including disulfiram, that exhibited effective activity against B. burgdorferi. In the current study, we evaluated the potential of repurposing the FDA-approved drug, disulfiram for its borreliacidal activity. Our results indicate disulfiram has excellent borreliacidal activity against both the log and stationary phase B. burgdorferi sensu stricto B31 MI. Treatment of mice with disulfiram eliminated the B. burgdorferi sensu stricto B31 MI completely from the hearts and urinary bladder by day 28 post infection. Moreover, disulfiram-treated mice showed reduced expressions of inflammatory markers, and thus they were protected from histopathology and cardiac organ damage. Furthermore, disulfiram-treated mice showed significantly lower amounts of total antibody titers (IgM and IgG) at day 21 and total IgG2b at day 28 post infection. FACS analysis of lymph nodes revealed a decrease in the percentage of CD19+ B cells and an increase in total percentage of CD3+ T cells, CD3+ CD4+ T helpers, and naive and effector memory cells in disulfiram-treated mice. Together, our findings suggest that disulfiram has the potential to be repurposed as an effective antibiotic for treating Lyme disease.

We developed a new approach for combined analysis of calcium (Ca2+) handling and beating forces in contractile cardiomyocytes. We employed human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) from dilated cardiomyopathy (DCM) patients carrying an inherited mutation in the sarcomeric protein troponin T (TnT), and isogenic TnT-KO iPSC-CMs generated via CRISPR/Cas9 gene editing. In these cells, Ca2+ handling as well as beating forces and-rates using single-cell atomic force microscopy (AFM) were assessed. We report impaired Ca2+ handling and reduced contractile force in DCM iPSC-CMs compared to healthy WT controls. TnT-KO iPSC-CMs display no contractile force or Ca2+ transients but generate Ca2+ sparks. We apply our analysis strategy to Ca2+ traces and AFM deflection recordings to reveal maximum rising rate, decay time, and duration of contraction with a multi-step background correction. Our method provides adaptive computing of signal peaks for different Ca2+ flux or force levels in iPSC-CMs, as well as analysis of Ca2+ sparks. Moreover, we report long-term measurements of contractile force dynamics on human iPSC-CMs. This approach enables deeper and more accurate profiling of disease-specific differences in cardiomyocyte contraction profiles using patient-derived iPSC-CMs.

We developed a new approach for combined analysis of calcium (Ca2+) handling and beating forces in contractile cardiomyocytes. We employed human induced pluripotent stem cell-derived cardiomyocytes (JPSC-CMs) from dilated cardiomyopathy (DCM) patients carrying an inherited mutation in the sarcomeric protein troponin T (TnT), and isogenic TnT-KO iPSC-CMs generated via CRISPR/Cas9 gene editing. In these cells, Ca2+ handling as well as beating forces and -rates using single-cell atomic force microscopy (AFM) were assessed. We report impaired Ca2+ handling and reduced contractile force in DCM iPSC-CMs compared to healthy WT controls. TnT-KO iPSC-CMs display no contractile force or Ca2+ transients but generate Ca2+ sparks. We apply our analysis strategy to Ca2+ traces and AFM deflection recordings to reveal maximum rising rate, decay time, and duration of contraction with a multi-step background correction. Our method provides adaptive computing of signal peaks for different Ca2+ flux or force levels in iPSC-CMs, as well as analysis of Ca2+ sparks. Moreover, we report long-term measurements of contractile force dynamics on human iPSC-CMs. This approach enables deeper and more accurate profiling of disease-specific differences in cardiomyocyte contraction profiles using patient-derived iPSC-CMs.

In vivo multiplexed imaging aims for noninvasive monitoring of tumors with multiple channels without excision of the tissue. While most of the preclinical imaging has provided a number of multiplexing channels up to three, Raman imaging with surface-enhanced Raman scattering (SERS) nanoparticles was suggested to offer higher multiplexing capability originating from their narrow spectral width. However, in vivo multiplexed SERS imaging is still in its infancy for multichannel visualization of tumors, which require both sufficient multiplicity and high sensitivity concurrently. Here we create multispectral palettes of gold multicore-near-infrared (NIR) resonant Raman dyes-silica shell SERS (NIR-SERRS) nanoparticle oligomers and demonstrate noninvasive and five-plex SERS imaging of the nanoparticle accumulation in tumors of living mice. We perform the five-plex ratiometric imaging of tumors by varying the administered ratio of the nanoparticles, which simulates the detection of multiple biomarkers with different expression levels in the tumor environment. Furthermore, since this method does not require the excision of tumor tissues at the imaging condition, we perform noninvasive and longitudinal imaging of the five-color nanoparticles in the tumors, which is not feasible with current ex vivo multiplexed tissue analysis platforms. Our work surpasses the multiplicity limit of previous preclinical tumor imaging methods while keeping enough sensitivity for tumor-targeted in vivo imaging and could enable the noninvasive assessment of multiple biological targets within the tumor microenvironment in living subjects.

CLARITY is a tissue preservation and optical clearing technique whereby a hydrogel is formed directly within the architectural confines of ex vivo brain tissue. In this work, the extent of polymer gel formation and crosslinking within tissue was assessed using Raman spectroscopy and rheology on CLARITY samples prepared with a range of acrylamide monomer (AAm) concentrations (1%, 4%, 8%, 12% w/v). Raman spectroscopy of individual neurons within hybrids revealed the chemical presence and distribution of polyacrylamide within the mouse hippocampus. Consistent with rheological measurements, lower %AAm concentration decreased shear elastic modulus G', providing a practical correlation with sample permeability and protein retention. Permeability of F(ab)'2 secondary fluorescent antibody changes from 9.3 to 1.4 mu m(2) s(-1) going from 1 to 12%. Notably, protein retention increased linearly relative to standard PFA-fixed tissue from 96.6% when AAm concentration exceeded 1%, with 12% AAm samples retaining up to similar to 99.3% native protein. This suggests that though 1% AAm offers high permeability, additional %AAm may be required to enhance protein. Our quantitative results on polymer distribution, stability, protein retention, and macromolecule permeability can be used to guide the design of future CLARITY-based tissue-clearing solutions, and establish protocols for characterization of novel tissue-polymer hybrid biomaterials using chemical spectroscopy and rheology.

[1] Crust and mantle discontinuities across the eastern margin of the North American craton were imaged using P to S converted phase receiver functions recorded by the Missouri to Massachusetts Broadband Seismometer Experiment. Crustal structure constrained by modeling Moho conversions and reverberations shows a variation of Moho depth from a minimum of 30 km near the Atlantic coast to depths of 44-49 km beneath the western Appalachian Province and 38-45 km beneath the Proterozoic terranes in the west. The variation in crustal thickness is substantially greater than that required for local isostasy, unless lower crustal densities are >3110 kg/m(3). In the upper mantle, Ps phases corresponding to a discontinuity at depths of 270-280 km were clearly observed beneath the eastern half of the array. Beneath the western third of the array, the receiver function stacks indicate more complex scattering, but weak Ps phases may be generated at depths of roughly 320 km. The transition between these two regions occurs across the eastern edge of the North American lithospheric keel imaged by tomography. The observed phases may be interpreted as conversions from the base of a low-velocity asthenosphere.

The ultraviolet-to-radio continuum spectral energy distributions are presented for all 75 galaxies in the Spitzer Infrared Nearby Galaxies Survey (SINGS). A principal component analysis of the sample shows that most of the sample's spectral variations stem from two underlying components, one representative of a galaxy with a low infrared-to-ultraviolet ratio and one representative of a galaxy with a high infrared-to-ultraviolet ratio. The influence of several parameters on the infrared-to-ultraviolet ratio is studied (e. g., optical morphology, disk inclination, far-infrared color, ultraviolet spectral slope, and star formation history). Consistent with our understanding of normal star-forming galaxies, the SINGS sample of galaxies in comparison to more actively star-forming galaxies exhibits a larger dispersion in the infrared-to-ultraviolet versus ultraviolet spectral slope correlation. Early-type galaxies, exhibiting low star formation rates and high optical surface brightnesses, have the most discrepant infrared-to-ultraviolet correlation. These results suggest that the star formation history may be the dominant regulator of the broadband spectral variations between galaxies. Finally, a new discovery shows that the 24 mu m morphology can be a useful tool for parameterizing the global dust temperature and ultraviolet extinction in nearby galaxies. The dust emission in dwarf/irregular galaxies is clumpy and warm accompanied by low ultraviolet extinction, while in spiral galaxies there is typically a much larger diffuse component of cooler dust and average ultraviolet extinction. For galaxies with nuclear 24 mu m emission, the dust temperature and ultraviolet extinction are relatively high compared to disk galaxies.

The Small Magellanic Cloud (SMC) provides a unique laboratory for the study of the lifecycle of dust given its low metallicity (similar to 1/5 solar) and relative proximity (similar to 60 kpc). This motivated the SAGE-SMC (Surveying the Agents of Galaxy Evolution in the Tidally Stripped, Low Metallicity Small Magellanic Cloud) Spitzer Legacy program with the specific goals of studying the amount and type of dust in the present interstellar medium, the sources of dust in the winds of evolved stars, and how much dust is consumed in star formation. This program mapped the full SMC (30 deg(2)) including the body, wing, and tail in seven bands from 3.6 to 160 mu m using IRAC and MIPS on the Spitzer Space Telescope. The data were reduced and mosaicked, and the point sources were measured using customized routines specific for large surveys. We have made the resulting mosaics and point-source catalogs available to the community. The infrared colors of the SMC are compared to those of other nearby galaxies and the 8 mu m/24 mu m ratio is somewhat lower than the average and the 70 mu m/160 mu m ratio is somewhat higher than the average. The global infrared spectral energy distribution (SED) shows that the SMC has approximately 1/3 the aromatic emission/polycyclic aromatic hydrocarbon abundance of most nearby galaxies. Infrared color-magnitude diagrams are given illustrating the distribution of different asymptotic giant branch stars and the locations of young stellar objects. Finally, the average SED of H II/star formation regions is compared to the equivalent Large Magellanic Cloud average H II/star formation region SED. These preliminary results will be expanded in detail in subsequent papers.

We present results of mid-infrared spectroscopic mapping observations of six star-forming regions in the Small Magellanic Cloud (SMC) from the Spitzer Spectroscopic Survey of the SMC ((SMC)-M-4). We detect the mid-IR emission from polycyclic aromatic hydrocarbons (PAHs) in all of the mapped regions, greatly increasing the range of environments where PAHs have been spectroscopically detected in the SMC. We investigate the variations of the mid-IR bands in each region and compare our results to studies of the PAH bands in the SINGS sample and in a sample of low-metallicity starburst galaxies. PAH emission in the SMC is characterized by low ratios of the 6-9 mu m features relative to the 11.3 mu m feature and weak 8.6 and 17.0 mu m features. Interpreting these band ratios in the light of laboratory and theoretical studies, we find that PAHs in the SMC tend to be smaller and less ionized than those in higher metallicity galaxies. Based on studies of PAH destruction, we argue that a size distribution shifted toward smaller PAHs cannot be the result of processing in the interstellar medium, but instead reflects differences in the formation of PAHs at low metallicity. Finally, we discuss the implications of our observations for our understanding of the PAH life-cycle in low-metallicity galaxies-namely that the observed deficit of PAHs may be a consequence of PAHs forming with smaller average sizes and therefore being more susceptible to destruction under typical interstellar medium conditions.

Topographic correction methods rarely consider the canopy parameter effects directly and explicitly for sloping canopies. In order to address this problem, the topographic correction method MFM-GOST2 was developed by implementing the second version of the Geometric-Optical model for Sloping Terrains (the GOST2 model) in the multiple forward mode (MFM) inversion framework. First, a look up table (LUT) was constructed by multiple forward modeling of the GOST2 model; second, the radiance of a remotely sensed image and its corresponding topographic data were used for searching potential canopy parameter combinations from the LUT; and third, the corrected radiance was determined by averaging potential radiances of horizontal canopies from the LUT according to the canopy parameter combinations. The MFM-GOST2 and twelve generally used topographic correction methods were evaluated via a case study by visual analysis, linear relationship analysis, and the rose diagram analysis. The result showed that the MFM-GOST2 method successfully removed most of the topographic effects of a subset image of the Landsat-8 image in a case study. The case study also illustrates that the rose diagram analysis is a good way to evaluate topographic corrections, but the linear relationship analysis cannot be used independently for the evaluations because the decorrelation is not a sufficient condition to determine a successful topographic correction.