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Novel Fret Method

Dr. Jiayu Liao and colleagues at the University of California, Riverside have developed a FRET assay using nitrobenzoxadiazole (NBD) and coumarin (CUM) amino acid analogs as a FRET pair.  These fluorophores are genetically encoded into peptides and proteins surrounding a protease cleavage site or ligand binding site and used for FRET-based high throughput screening for enzymes or small molecule inhibitors involved in pathways such as SUMOylation. Researchers have demonstrated FRET for peptides encoded with NBD and CUM separated by 4 and 6 amino acids and excited at 340 nm (Figure 1). Figure 1.  Fluorescent intensity of peptide I (6 amino acids between CUM and NBD) and II (4 amino acids between CUM and NBD) excited at 340 nm.  

New Catalysts for Perchlorate Reduction in Water

Prof. Jinyong Liu’s lab at UCR has developed a new family of catalysts that reduce perchlorate in contaminated water and wastewater. The catalyst rapidly and completely reduces the toxic ClO4- into the innocuous chloride (Cl -) by breaking down the bonds between the central chlorine atom and all surrounding oxygen atoms. The reduction is a green process because no byproducts are produced in the water. The catalyst completely reduces perchlorate in a very wide concentration range, and retains high activity even in brine with concentrated salts. The catalyst using earth-abundant and non-toxic metal provides sustainable solutions to the perchlorate issues in terms of water and wastewater treatment, ion-exchange resin regeneration, and old munition/explosive disposal. Not only can this new catalyst reduce perchlorate but it may also be used to reduce other drinking water contaminants such as chlorate, chlorite, nitrate, nitrite, bromate, and iodate in a variety of environmental remediation scenarios.  Fig. 1 shows the reduction profiles of 1, 10, and 100 mM ClO4− (corresponding to 100,000 to 10,000,000 ppb) by the UCR catalyst at a loading of only 0.2 g/L. The reactions were conducted at 25 oC and under 1 atm H2. Fig. 2 shows the high activity for the catalytic reduction of 1 mM ClO4− by the UCR catalyst (just 0.2 g/L) in the typical resin generation wastes containing chloride and sulfate.

Chronoprints: Identifying Adulterated Samples in Food and Drug Safety

Prof. Will Grover and his colleague at the University of California have developed a method to identify adulterated drugs and foods by observing how they behave when disturbed by temperature changes or other causes. Images of the sample’s behavior as it freezes over time are captured and processed into chronoprints.  Chronoprints are fundamentally bitmap images of samples on a computer, and it is possible to leverage existing image analysis and comparison techniques that have been already developed to analyze Chronoprints. Fig. 1 Producing a "chronological fingerprint" or chronoprint capturing how six samples (in this example, authentic and adulterated samples of an over-the-counter liquid cold medicine) respond to a perturbation over space and time (in this case, a rapidly changing temperature gradient). (A) A microfluidic thermometer chip containing the samples is partially immersed in liquid nitrogen to establish a rapidly changing temperature gradient along the chip. (B) The chip contains six samples (red) loaded in microfluidic channels that run parallel to the dynamic temperature gradient. (C) An inexpensive USB microscope records a video of the physical changes in the samples as they react to the dynamic temperature gradient.  Fig. 2 By reducing each channel image to a single column of pixels, and then placing these columns side-by-side, we create a bitmap image (the sample’s chronoprint) that captures how the sample changes over space (the y-axis) and time (the x-axis). Finally, by comparing the chronoprints of all six samples in the chip, we can determine whether the samples are either likely the same or definitely different.  

Accurate and Secure Navigation for Autonomous Vehicles

While cellular phone networks are not designed for navigation, they are abundant in urban environments which are known to challenge GPS signals.  University of California, Riverside researchers integrated signals-of-opportunity from mobile phone networks to provide autonomous vehicles with precise navigational information.

Modified Enzymes to Improve Crop Yield

Researchers at the University of California have identified new modified versions of the carbon fixing enzyme, Phosphoenolpyruvate carboxylase (PPC).  in planta results show that the modified PPC enzymes confer upwards of a five fold increase in carbon fixation when compared to wild type plants. PPC dependent carbon fixation is key to photosynthesis, production of nutrients, and plants conditioning their growth environment. Plants with modified PPCs that increase carbon fixation and photosynthetic output will have increased plant productivity, which is critical for feeding a growing population. Additionally, by identifying surgical changes that can unleash the full productivity of plant PPC’s, it will be possible to increase the rate of depletion of atmospheric CO2.  The combination of these outcomes represents the opportunity to boost agricultural productivity, increase the amount of agriculturally available land by upwards of 100%, and improve the nutritional quality of plants all of which are dependent on removal of CO2 from our atmosphere.  Fig. 2 in vitro comparison of wild type (wt) and modified versions of maize PPC1, which is key to C4 photosynthesis, in the absence or presence of increasing amounts of the allosteric inhibitor, malate. Whereas version A is less affected by malate than wt, both versions B and C are largely unaffected by malate and have a 2-fold increase in activity compared to the wt version.  

Novel Steroid Hormone Assay

Researchers at the University of California have identified in insects that the membrane transporter, Ecdysone Importer (EcI), is involved in the cellular uptake of the primary steroid hormone ecdysone. Specifically after transport through Ecl, ecdysone’s active form (20-hydroxyecdysone or 20E and related ecdysteroids) enters its target cells and binds to the ecdysone receptor (EcR), which forms a heterodimer with another nuclear receptor and activates transcription of multiple genes involved in molting and metamorphosis. This new discovery of Ecl’s role counters the prevailing consensus that steroid hormones diffuse through cell membranes.  This will enable the screening of new compounds that interact with Ecl.  Such new compounds may be used for insect pest control. Fig. 1 membrane transporters (blue) guide steroid hormones (blue dots) into cells. This new discovery counters the conventionally held scientific consensus that steroid hormones passively diffuse through cell membranes.   Fig. 2 EcI mutants (bottom) were not able to enter into metamorphosis when compared to the control (top).

Method to Reuse Multielectrode Arrays in Rodents

Researchers at the University of California have developed a protocol to enable the reuse of MEA probes.  Using this protocol, the MEA probes can be carefully peeled off undamaged from a protective layer, cleaned with ethanol and stored for re-use.  In addition, at each reuse the measured electrode impedances remain within the normal range set by the manufacturer for every channel and the probes may be reused up to six times.  This protocol is an improvement over the existing published protocols in that (1) these particular MEA electrodes are available commercially in a variety of configurations; (2) the MEA can be reused a number of times in order to record EEG in freely moving mice. Fig. 2 Setup of MEA EEG that allowed for enhanced reusability.

Plants Resistant to Fungal Disease

University of California, Riverside researcher Prof. Hailing Jin and her colleagues have developed plants that are resistant to Botrytis cineria and Verticillium dahlia. These plants are genetically engineered to silence fungal pathogens that transfer “virulent” small RNA effectors to the plant that cause disease.  This has led to the development of plants that are resistant to Botrytis cineria and Verticillium dahlia. Fig. 1 shows fruits (bottom) with dramatic reductions in gray mold disease. Gray mold disease is caused by Botrytis cineria. The bottom fruits were sprayed with small RNA (sRNA) against Botrytis cineria pathogens dicer-like 1 & 2 (BcDCL). The top fruits were sprayed with water and this conferred no protection against gray mold disease. Immunity to pathogens may be genetically engineered into plants to express BcDCL-1 and BcDCL-2.