Purification and functional reconstitution of human olfactory receptor expressed in Escherichia coli

Olfactory receptors (ORs), belonging to the Gprotein coupled receptor (GPCR) family, are very difficult to be overexpressed, purified and reconstituted because of their hydrophobicity and complicated structure. These receptors bind to their specific ligands, thus their specificity is very useful for application as a bioelectronic nose. Furthermore, highly purified and well-reconstituted human olfactory receptor (hOR) can be used in various fields, such as in protein-interaction research, drug screening, and analysis of the hOR structure. In this study, human olfactory receptor, hOR2AG1, was produced with high purity and functionally reconstituted in detergent micelles. The hOR2AG1 was overexpressed in Escherichia coli (E. coli) with glutathione S-transferase (GST) and 6xHis-tag as an inclusion body. The hOR2AG1 fusion protein was solubilized in buffer containing sodium dodecyl sulfate (SDS) and purified using Ni-NTA chromatography. The GST domain was removed using proteolytic cleavage before elution from the column. After purification, the hOR2AG1 was successfully reconstituted using nonionic detergents and methyl-ß-cyclodextrin. Finally highly purified and well-reconstituted hOR was obtained, and its biological characteristics were confirmed by using circular dichroism (CD) spectrum and tryptophan fluorescence assay. These results can be applied to develop protein-based sensing systems including a bioelectronic nose and to analyze the native hOR structure using solid-state NMR, X-ray crystallography, or neutron scattering.Korea (South). Ministry of Science, ICT and Future Planning (Grants2014039771 and 2014053108)Korea Institute of Science and Technology (KIST) (Project 2E24812-14-043

Down-regulation of phosphoglucomutase 3 mediates sulforaphane-induced cell death in LNCaP prostate cancer cells

<p>Abstract</p> <p>Background</p> <p>Sulforaphane (SFN) is an isothiocyanate found in cruciferous vegetables that exerts anti-oxidant, anti-inflammatory, anti-cancer and radio-sensitizing activities. Nonetheless, the mechanism responsible for SFN-induced cell death is not fully understood. In the present study, anti-cancer mechanism of SFN was elucidated in LNCaP prostate cancer cells.</p> <p>Results</p> <p>SFN exerted cytotoxicity and increased TUNEL positive cells in a concentration-dependent manner in LNCaP cells. Proteomics study revealed that levels of nine proteins including tubulin β-2, phosphoglucomutase-3 (PGM3), melanoma-derived leucine zipper containing extra-nuclear factor, activin A type I receptor precursor, smoothelin-A, KIA0073, hypothetical protein LOC57691 and two unnamed proteins were changed over 8 folds in SFN treated LNCaP cells compared to untreated control. We have further confirmed that SFN reduced PGM3 expression with western blotting and showed that PGM3 siRNA enhanced cytotoxicity demonstrated by cell morphology and TUNEL assays in LNCaP cells.</p> <p>Conclusion</p> <p>Taken together, these findings suggest that PGM3 plays a role in mediating SFN-induced cell death in LNCaP cells, and is a potential molecular therapeutic target for prostate cancer.</p

Human dopamine receptor nanovesicles for gate-potential modulators in high-performance field-effect transistor biosensors

The development of molecular detection that allows rapid responses with high sensitivity and selectivity remains challenging. Herein, we demonstrate the strategy of novel bio-nanotechnology to successfully fabricate high-performance dopamine (DA) biosensor using DA Receptor-containing uniform-particle-shaped Nanovesicles-immobilized Carboxylated poly(3,4-ethylenedioxythiophene) (CPEDOT) NTs (DRNCNs). DA molecules are commonly associated with serious diseases, such as Parkinson's and Alzheimer's diseases. For the first time, nanovesicles containing a human DA receptor D1 (hDRD1) were successfully constructed from HEK-293 cells, stably expressing hDRD1. The nanovesicles containing hDRD1 as gate-potential modulator on the conducting polymer (CP) nanomaterial transistors provided high-performance responses to DA molecule owing to their uniform, monodispersive morphologies and outstanding discrimination ability. Specifically, the DRNCNs were integrated into a liquid-ion gated field-effect transistor (FET) system via immobilization and attachment processes, leading to high sensitivity and excellent selectivity toward DA in liquid state. Unprecedentedly, the minimum detectable level (MDL) from the field-induced DA responses was as low as 10 pM in real- time, which is 10 times more sensitive than that of previously reported CP based-DA biosensors. Moreover, the FET-type DRNCN biosensor had a rapid response time (<1 s) and showed excellent selectivity in human serum

A High-Speed Range-Matching TCAM for Storage-Efficient Packet Classification

Abstract—A critical issue in the use of TCAMs for packet classification is how to efficiently represent rules with ranges, known as range matching. A range-matching ternary content addressable memory (RM-TCAM) including a highly functional range-matching cell (RMC) is presented in this paper. By offering various range operators, the RM-TCAM can reduce storage expansion ratio from 4.21 to 1.01 compared with conventional TCAMs, under real-world packet classification rule sets, which results in reduced power consumption and die area. A new pre-discharging match-line scheme is used to realize high-speed searching in a dynamic match-line structure. An additional charge-recycling driver further reduces the power consumption of search lines. Simulation results of a 256 64-bit range-matching TCAM, when implemented in the 0.13- m CMOS technology, achieves a 1.99-ns search time with an energy efficiency of 1.26 fJ/bit/search. While a TCAM including range encoding approach requires an additional SRAM or DRAM, the RM-TCAM can improve storage efficiency without any extra components as well as reduce the die area

Breaking the absorption limit of Si toward SWIR wavelength range via strain engineering

Silicon has been widely used in the microelectronics industry. However, its photonic applications are restricted to visible and partial near-infrared spectral range owing to its fundamental optical bandgap (1.12 eV). With recent advances in strain engineering, material properties, including optical bandgap, can be tailored considerably. This paper reports the strain-induced shrinkage in the Si bandgap, providing photosensing well beyond its fundamental absorption limit in Si nanomembrane (NM) photodetectors (PDs). The Si-NM PD pixels were mechanically stretched (biaxially) by a maximum strain of similar to 3.5% through pneumatic pressure-induced bulging, enhancing photoresponsivity and extending the Si absorption limit up to 1550 nm, which is the essential wavelength range of the lidar sensors for obstacle detection in self-driving vehicles. The development of deformable three-dimensional optoelectronics via gas pressure-induced bulging also facilitated the realization of unique device designs with concave and convex hemispherical architectures, which mimics the electronic prototypes of biological eyes.1

Pseudoinvasion in an Adenomatous Polyp of the Colon Mimicking Invasive Colon Cancer

Pseudoinvasion or pseudocarcinomatous invasion in an adenomatous polyp of the colon can be unfamiliar to an endoscopist. Pseudoinvasion in an adenomatous polyp represents prolapse of the adenomatous epithelium into its stalk. In most cases its morphology does not differ from of general adenomatous polyps, but in some cases it can morphologically mimic a malignant polyp with submucosal invasion due to mass-like lesioning of its stalk. This makes it difficult for endoscopists to differentiate pseudoinvasion in an adenoma from an invasive carcinoma by conventional endoscopy; instead, endoscopic ultrasonography can provide useful information for differentiating these conditions. We report on an 82-year-old man who presented with a large pedunculated polyp with a thick stalk in the sigmoid colon, which mimicked a submucosal invasive carcinoma. The patient was diagnosed with pseudoinvasion in an adenomatous polyp after segmental resection of the sigmoid colon

N=8 SCFT and M Theory on AdS_4 x RP^7

We study M theory on AdS_4 \times \RP^7 corresponding to 3 dimensional ${\cal N}=8$ superconformal field theory which is the strong coupling limit of 3 dimensional super Yang-Mills theory. For SU(N) theory, a wrapped M5 brane on \RP^5 can be interpreted as baryon vertex. For $SO(N)/Sp(2N)$ theory, by using the property of (co-)homology of \RP^7, we classify various wrapping branes and consider domain walls and the baryon vertex.Comment: 17 pages, Changed baryon like operator as M5 branes in M theory rather than D6 brane in IIA theory. To appear in Phys.Rev.

Chlorophyll and Total Suspended Materials Concentrations and Remote Sensing Reflectance Data measured at the Red Tide Area of Jinhae, Geoje, and East Sea during August from 1998 to 2003 and August 2013

The chlorophyll and total suspended materials concentrations and remote sensing reflectance data were observed for red tides occurring every summer in waters around the Korean Peninsula. In observation area and date, the field survey were performed (1) in the Jinhae and Geoje coasts during August 1998, August 1999, August 2001, and August 2003, (2) in East Sea coast during August 2013. The remote sensing reflectance data were obtained from portable spectroradiometer. The chlorophyll concentration data were obtained from spectrophotometric method and the total suspended materials concentration data were obtained from filter-weight difference method. The remote sensing reflectance data were validated using Moon et al.(2012). The chlorophyll concentration data were validated using baseline correction and subtraction of 750 nm value, and the total suspended materials concentration data were validated using variation of humidity