ABSTRACT: This experiment tests the effect of individual mutations on the metabolome of Arabidopsis. The standardized growth conditions are as follows: 1. Seeds (between 14 and 16) are sown on media in 100 x 100 x 15mm square Falcon Petri Dishes (Fisher Scientific, catalogue #08-757-11A). Seeds were arranged on the plates in a single horizontal line at the 1-cm mark from the top of the plate. 2. Each plate contains between 20 and 25-ml of sterile MS media containing 0.1% (w/v) sucrose. 3. Prior to sowing, seeds were sterilized by treating for 1-minute at room temperature with a 300-l solution of 50% (v/v) ethanol, this solution was removed and replaced with a 300-l solution consisting of 1% (v/v) Tween 20 (Fischer BioReagents, catalogue #BP33750), and 50% (v/v) bleach solution (Clorox), and incubated at room temperature for 10-minutes. The seeds were then washed with three changes of 0.3-ml of sterile water. 4. Upon sowing with seeds, the plates were wrapped with Micropore tape (3M Health Care, catalogue #1530-0), and then stored horizontally for 4-days at 4 °C, with illumination of 1 mol/m2. 5. On the 5th day, plates were moved to the growth room, and held in a vertical position in Plexi-glass holders for 17-days this growth room condition is labeled in Table I. 6. On 18th day Petri plates were opened and the aerial portions of these plants were harvested immediately upon plate opening. 7. Upon harvesting, plant material was quenched by immersion in liquid nitrogen and stored at 70 °C.
Project description:This experiment tests the consequence of a mutation at the FatB gene (At1g08510) in the wound-response of Arabidopsis. The FatB mutant allele (fatb KD J. Ohlrogge (Plant Cell 2003, Vol 15, 1020-1033)) was obtained from Dr. Katayonn Dehesh, University of California, Davis, Davis, CA. This allele is in the Ws background.The standardized growth conditions are as follows: 1. Seeds (between 14 and 16) are sown on media in 100 x 100 x 15mm square Falcon Petri Dishes (Fisher Scientific, catalogue #08-757-11A). Seeds were arranged on the plates in a single horizontal line at the 1-cm mark from the top of the plate.2. Each plate contains between 20 and 25-ml of sterile MS media containing 0.1% (w/v) sucrose.3. Prior to sowing, seeds were sterilized by treating for 1 minute at room temperature with a 300-l solution of 50% (v/v) ethanol, this solution was removed and replaced with a 300-l solution consisting of 1% (v/v) Tween 20 (Fischer BioReagents, catalogue #BP33750), and 50% (v/v) bleach solution (Clorox), and incubated at room temperature for 10-minutes. The seeds were then washed with three changes of 0.3-ml of sterile water.
Project description:Effect of microwave heating on the crystallization of glutathione (GSH) tripeptide using the metal-assisted and microwave-accelerated evaporative crystallization (MA-MAEC) technique is reported. GSH crystals were grown from supersaturated solutions of GSH (300-500 mg/mL) on the iCrystal plates with silver nanoparticle films (SNFs) and without SNFs in three different microwave systems operating at 2.45 GHz: conventional (multimode, fixed power at 900W), industrial (monomode, variable power up to 1200 W), and the iCrystal system (monomode, variable power up to 100 W). The efficacy of the MA-MAEC technique, in terms of improvement in the crystallization time, crystal size and quality of GSH, was compared between the three microwave systems and the crystallization at room temperature (no microwave heating, a control experiment). Optical microscopy was used to visualize and quantify the growth of GSH crystals during and after microwave heating. Powder X-ray diffraction (XRD) and Fourier transform infrared (FTIR) spectroscopy data showed that GSH crystals had identical crystal structure to those grown at room temperature and microwave heating did not alter the chemical structure of GSH molecules during microwave heating, respectively. Using the MA-MAEC technique, the iCrystal system yielded high quality GSH crystals in a rapid manner.
Project description:Copper nanoparticles (NPs) with an average particle diameter of 50-60?nm were successfully obtained by reducing an aqueous solution of a copper(II)-nitrilotriacetic acid complex with an aqueous hydrazine solution at room temperature under an air atmosphere. Copper NP-based nanopastes were printed onto a glass substrate using a metal screen mask and pressureless sintered under a nitrogen atmosphere at 200?°C for 30?min. The electrical resistivity of the resulting copper electrode was 16 ???·?cm. For a metal-to-metal bonding test, copper nanopaste was printed on an oxygen-free copper plate, another oxygen-free copper plate was placed on top, and the bonding strength between the copper plates when pressureless sintered under a nitrogen atmosphere at 200?°C for 30?min was 39?MPa. TEM observations confirmed that highly crystalline metal bonding occurred between the copper NPs and the copper plate to introduce the ultrahigh strength. The developed copper NPs could provide promising advances as nanopastes for sustainable fabrication of copper electrodes and die attachment materials for the production of next-generation power semiconductors.
Project description:Despite significant progress in high-throughput methods in macromolecular crystallography, the production of diffraction-quality crystals remains a major bottleneck. By recording diffraction in situ from crystals in their crystallization plates at room temperature, a number of problems associated with crystal handling and cryoprotection can be side-stepped. Using a dedicated goniometer installed on the microfocus macromolecular crystallography beamline I24 at Diamond Light Source, crystals have been studied in situ with an intense and flexible microfocus beam, allowing weakly diffracting samples to be assessed without a manual crystal-handling step but with good signal to noise, despite the background scatter from the plate. A number of case studies are reported: the structure solution of bovine enterovirus 2, crystallization screening of membrane proteins and complexes, and structure solution from crystallization hits produced via a high-throughput pipeline. These demonstrate the potential for in situ data collection and structure solution with microbeams.
Project description:This in vitro study was conducted to evaluate the shear bond strength of "non-self-adhesive" resin to dental zirconia etched with hydrofluoric acid (HF) at room temperature and to compare it to that of air-abraded zirconia. Sintered zirconia plates were air-abraded (control) or etched with 10%, 20%, or 30% HF for either 5 or 30 min. After cleaning, the surfaces were characterized using various analytical techniques. Three resin cylinders (Duo-Link) were bonded to each treated plate. All bonded specimens were stored in water at 37 °C for 24 h, and then half of them were additionally thermocycled 5000 times prior to the shear bond-strength tests (n = 12). The formation of micro- and nano-porosities on the etched surfaces increased with increasing concentration and application time of the HF solution. The surface wettability of zirconia also increased with increasing surface roughness. Higher concentrations and longer application times of the HF solution produced higher bond-strength values. Infiltration of the resin into the micro- and nano-porosities was observed by scanning electron microscopy. This in vitro study suggests that HF slowly etches zirconia ceramic surfaces at room temperature, thereby improving the resin-zirconia bond strength by the formation of retentive sites.
Project description:AuNPs ranging in 20 to 300 nm size were synthesized at a room temperature using Yucca filamentosa leaf extract. Diverse nanomaterial morphologies were obtained by varying the extract concentration, reaction pH, and temperature. While low volumes of extract (0.25 and 0.5 mL) induced the formation of microscale Au sheets with edge length greater than 1 ?m, high volumes yielded spherical particles ranging from 20 to 200 nm. Varying pH of the solution significantly influenced the particle shape with the production of largely spherical particles at pH 5 to 6 and truncated triangles at pH 2. Separation of multidimensional nanostructures was achieved using a novel method of sucrose density gradient centrifugation. The catalytic function of Yucca-derived AuNPs was demonstrated by degradation of a wastewater dye: methylene blue using spectrophotometric measurements over time. Treatment with Au nanosheets and spheres demonstrated methylene blue degradation approximately 100% greater than the activity in control at 60 min.
Project description:Ambient temperature and physical activity modulate bone elongation in mammals, but mechanisms underlying this plasticity are a century-old enigma. Longitudinal bone growth occurs in cartilaginous plates, which receive nutritional support via delivery of solutes from the vasculature. We tested the hypothesis that chronic exercise and warm temperature promote bone lengthening by increasing solute delivery to the growth plate, measured in real time using in vivo multiphoton microscopy. We housed 68 weanling female mice at cold (16°C) or warm (25°C) temperatures and allowed some groups voluntary access to a running wheel. We show that exercise mitigates the stunting effect of cold temperature on limb elongation after 11 days of wheel running. All runners had significantly lengthened limbs, regardless of temperature, while nonrunning mice had shorter limbs that correlated with housing temperature. Tail length was impacted only by temperature, indicating that the exercise effect was localized to limb bones and was not a systemic endocrine reaction. In vivo multiphoton imaging of fluoresceinated tracers revealed enhanced solute delivery to tibial growth plates in wheel-running mice, measured under anesthesia at rest. There was a minimal effect of rearing temperature on solute delivery when measured at an intermediate room temperature (20°C), suggesting that a lasting increase in solute delivery is an important factor in exercise-mediated limb lengthening but may not play a role in temperature-mediated limb lengthening. These results are relevant to the study of skeletal evolution in mammals from varying environments and have the potential to fundamentally advance our understanding of bone elongation processes.
Project description:Light transmission aggregometry (LTA) can be performed with microtiter plates (96-well LTA). When conducting LTA, an agonist is added to platelet-rich plasma and the sample is shaken for minutes after which absorbance readings are done. Platelet aggregation is detected as decrease in absorbance. However, the classical method is cumbersome and therefore microtiter plates can be used for concomitant testing of multiple samples. Furthermore, it would be convenient to prepare the plate in advance of platelet aggregation testing. Aim: The aim of the present study was to establish a simplified 96-well LTA protocol, where plates were pre-coated with agonists and stored at -80 C until use.We developed and validated a protocol for 96-well LTA using a Victor X5 plate reader and pre-coated microtiter plates. The minimum requirement of platelet-rich plasma was 45 ?L per sample and the sample platelet count should not be below 100 x109/L. Optimal absorbance reading was 595 nm wavelengths. Platelet aggregation results were higher at 37°C than at room temperature. Platelet adherence to wells after stimulation was observed; it was not avoided by pre-coating of the wells with gelatin. A range of up to 7 concentrations for each agonist (collagen, arachidonic acid, adenosine diphosphate, thrombin receptor-activating peptide and protease-activated receptor-4) was tested concomitantly. A transient rise in platelet aggregation was observed after 2 minutes of shaking in some samples with low agonist concentration, and platelet aggregation was optimal after 10 minutes of shaking for samples with high agonist concentration. Plates could be stored at -80°C for 15 days without significant change in the platelet aggregation results.The 96-well LTA is suitable for platelet aggregation testing and a range of agonist concentrations can be concomitantly tested.
Project description:In this paper, ultrasonic-enhanced replacement of lead by zinc in lead leaching solution was studied. The effects of reaction time, rotational speed, temperature, concentration of leaching solution and the ratio of the surface area of the zinc plate immersed in the leaching solution to the volume of leaching solution (S : V) were studied under both conventional and ultrasonic conditions. The optimum ultrasonic-assisted replacement conditions were as follows: the S : V of 0.04 (4 cm2 100 ml-1), reaction temperature of 30°C, replacement time of 30 min and the concentration of leaching solution is 5 g l-1, leading to a lead replacement rate of 94.84%. Compared with the conventional replacement process, the reaction time of ultrasonic-enhanced substitution could be reduced to one half, and the demand of reaction temperature, leaching solution concentration and other conditions were decreased accordingly. Introducing ultrasonic into the replacement reaction is promising to reduce the energy consumption in the hydrometallurgical industry also caters to the demands of environment protection.
Project description:The refractory tungsten alloys with high ductility/strength/plasticity are highly desirable for a wide range of critical applications. Here we report an interface design strategy that achieves 8.5?mm thick W-0.5?wt. %ZrC alloy plates with a flexural strength of 2.5?GPa and a strain of 3% at room temperature (RT) and ductile-to-brittle transition temperature of about 100?°C. The tensile strength is about 991?MPa at RT and 582?MPa at 500?°C, as well as total elongation is about 1.1% at RT and as large as 41% at 500?°C, respectively. In addition, the W-ZrC alloy plate can sustain 3.3?MJ/m(2) thermal load without any cracks. This processing route offers the special coherent interfaces of grain/phase boundaries (GB/PBs) and the diminishing O impurity at GBs, which significantly strengthens GB/PBs and thereby enhances the ductility/strength/plasticity of W alloy. The design thought can be used in the future to prepare new alloys with higher ductility/strength.