Nicotinic acid (niacin) has been widely used as a favorable lipid-lowering drug for several decades, and the orphan G protein-coupled receptor GPR109A has been identified to be a receptor for niacin. Mechanistic investigations have shown that as a G i -coupled receptor, GPR109A inhibits adenylate cyclase activity upon niacin activation, thereby inhibiting free fatty acid liberation. However, the underlying molecular mechanisms that regulate signaling and internalization of GPR109A remain largely unknown. To further characterize GPR109A internalization, we made a construct to express GPR109A fused with enhanced green fluorescent protein (EGFP) at its carboxyl-terminal end. In stable GPR109A-EGFP-expressing HEK-293 cells, GPR109A-EGFP was mainly localized at the plasma membrane and was rapidly internalized in a dose-and time-dependent manner upon agonist stimulation. GPR109A internalization was completely blocked by hypertonic sucrose, indicating that GPR109A internalizes via the clathrin-coated pit pathway. Further investigation demonstrated that internalized GPR109A was recycled to the cell surface after the removal of agonist, and recycling of the internalized receptors was not blocked by treatment with acidotropic agents, NH 4 Cl and monensin. Pertussis toxin pretreatment not only inhibited forskolin-induced cAMP accumulation and intracellular Ca 2؉ mobilization; it also significantly attenuated agonist-promoted GPR109A internalization. Moreover, RNA interference experiments showed that knockdown of GRK2 (G protein-coupled receptor kinase 2) and arrestin3 expression significantly impaired receptor internalization. Taken together, these results indicate that the agonist-induced internalization of GPR109A receptors is regulated by GRK2 and arrestin3 in a pertussis toxin-sensitive manner and that internalized receptor recycling is independent of endosomal acidification.

We applied a linear-array-based photoacoustic probe to detect melanin-containing melanoma tumor depth and volume in nude mice in vivo. This system can image melanomas at five frames per second (fps), which is much faster than our previous handheld single transducer system (0.1 fps). We first theoretically show that, in addition to the higher frame rate, almost the entire boundary of the melanoma can be detected by the linear-array-based probe, while only the horizontal boundary could be detected by the previous system. Then we demonstrate the ability of this linear-array-based system in measuring both the depth and volume of melanoma through phantom, ex vivo, and in vivo experiments. The volume detection ability also enables us to accurately calculate the rate of growth of the tumor, which is an important parameter in quantifying the tumor activity. Our results show that this system can be used for clinical melanoma diagnosis and treatment in humans at the bedside.

Abstract. Capitalizing on endogenous hemoglobin contrast, photoacoustic-computed tomography (PACT), a deep-tissue high-resolution imaging modality, has drawn increasing interest in neuroimaging. However, most existing studies are limited to functional imaging on the cortical surface and the deep brain structural imaging capability of PACT has never been demonstrated. Here, we explicitly studied the limiting factors of deep brain PACT imaging. We found that the skull distorted the acoustic signal and blood suppressed the structural contrast from other chromophores. When the two effects are mitigated, PACT can potentially provide high-resolution label-free imaging of structures in the entire mouse brain. With 100-μm in-plane resolution, we can clearly identify major structures of the brain, which complements magnetic resonance microscopy for imaging small-animal brain structures. Spectral PACT studies indicate that structural contrasts mainly originate from cytochrome distribution and that the presence of lipid sharpens the image contrast; brain histology results provide further validation. The feasibility of imaging the structure of the brain in vivo is also discussed. Our results demonstrate that PACT is a promising modality for both structural and functional brain imaging.

Figure 1: Given a closed 2-manifold mesh, we compute a scalar field (a), which accentuates the axes of prominent, partial intrinsic reflectional symmetries. The top few (closed) Voronoi boundaries (b) between symmetric point pairs, as induced by the scalar field, can be imperfect. We develop an iterative refinement scheme to extract the final set of intrinsic reflectional symmetry axes or IRSAs (c), which can be open curves. Incorporating symmetry cues offered by IRSAs into a conventional mesh segmentation scheme leads to highly semantic results (d). AbstractWhile many 3D objects exhibit various forms of global symmetries, prominent intrinsic symmetries which exist only on parts of an object are also well recognized. Such partial symmetries are often seen as more natural than a global one, even when the symmetric parts are under complex pose. We introduce an algorithm to extract partial intrinsic reflectional symmetries (PIRS) of a 3D shape. Given a closed 2-manifold mesh, we develop a voting scheme to obtain an intrinsic reflectional symmetry axis (IRSA) transform, which is a scalar field over the mesh that accentuates prominent IRSAs of the shape. We then extract a set of explicit IRSA curves on the shape based on a refined measure of local reflectional symmetry support along a curve. The iterative refinement procedure combines IRSA-induced region growing and region-constrained symmetry support refinement to improve accuracy and address potential issues arising from rotational symmetries in the shape. We show how the extracted IRSA curves can be incorporated into a conventional mesh segmentation scheme so that the implied symmetry cues can be utilized to obtain more meaningful results. We also demonstrate the use of IRSA curves for symmetry-driven part repair.