We have previously used cyclic nucleotide-gated (CNG) channels as sensors to measure cAMP signals in human embryonic kidney (HEK)-293 cells. We found that prostaglandin E(1) (PGE(1)) triggered transient increases in cAMP concentration near the plasma membrane, whereas total cAMP levels rose to a steady plateau over the same time course. In addition, we presented evidence that the decline in the near-membrane cAMP levels was due primarily to a PGE(1)-induced stimulation of phosphodiesterase (PDE) activity, and that the differences between near-membrane and total cAMP levels were largely due to diffusional barriers and differential PDE activity. Here, we examine the mechanisms regulating transient, near-membrane cAMP signals. We observed that 5-min stimulation of HEK-293 cells with prostaglandins triggered a two- to threefold increase in PDE4 activity. Extracellular application of H89 (a PKA inhibitor) inhibited stimulation of PDE4 activity. Similarly, when we used CNG channels to monitor cAMP signals we found that both extracellular and intracellular (via the whole-cell patch pipette) application of H89, or the highly selective PKA inhibitor, PKI, prevented the decline in prostaglandin-induced responses. Following pretreatment with rolipram (a PDE4 inhibitor), H89 had little or no effect on near-membrane or total cAMP levels. Furthermore, disrupting the subcellular localization of PKA with the A-kinase anchoring protein (AKAP) disruptor Ht31 prevented the decline in the transient response. Based on these data we developed a plausible kinetic model that describes prostaglandin-induced cAMP signals. This model has allowed us to quantitatively demonstrate the importance of PKA-mediated stimulation of PDE4 activity in shaping near-membrane cAMP signals.

Cyclic nucleotide-gated (CNG) channels are the primary targets of light-and odorant-induced signaling in photoreceptors and olfactory sensory neurons. Compartmentalized cyclic nucleotide signaling is necessary to ensure rapid and efficient activation of these nonselective cation channels. However, relatively little is known about the subcellular localization of CNG channels or the mechanisms of their membrane partitioning. Lipid raft domains are specialized membrane microdomains rich in cholesterol and sphingolipids that have been implicated in the organization of many membrane-associated signaling pathways. Herein, we report that the ␣ subunit of the olfactory CNG channel, CNGA2, associates with lipid rafts in heterologous expression systems and in rat olfactory epithelium. However, CNGA2 does not directly bind caveolin, and its membrane localization overlaps only slightly with that of caveolin at the surface of human embryonic kidney (HEK) 293 cells. To test for a possible functional role of lipid raft association, we treated HEK 293 cells with the cholesterol-depleting agent, methyl-␤-cyclodextrin. Cholesterol depletion abolished prostaglandin E 1 -stimulated CNGA2 channel activity in intact cells. Recordings from membrane patches excised from CNGA2-expressing HEK 293 cells revealed that cholesterol depletion dramatically reduced the apparent affinity of homomeric CNGA2 channels for cAMP but only slightly reduced the maximal current. Our results show that olfactory CNG channels target to lipid rafts and that disruption of lipid raft microdomains dramatically alters the function of CNGA2 channels.

Metallic powder has applications in many fields. In applications for preservation and anti-oxidation, iron powder has been used as an air oxygen reducer which is capable of decreasing microclimatic oxygen concentrations in a hermetic mini-environment. In this role, if we increase the specific surface area by reducing the particle size of the iron powder, the rate and performance of oxygen reduction will be improved significantly. In addition, the porosity of iron powder also contributes considerably. The iron powder can be fabricated using many methods: chemical deposition, powder metallurgy and mechanical milling. The technique of milling has certain advantages, especially for the formation of technical iron powder. The experimental equipment used was a Fritsch P-6 planetary ball mill. The iron powder was prepared with different milling times, from 1 up to 30 h in acetone as a protective environment. The powder products obtained were analyzed using field emission scanning electron microscope (FE-SEM), energy dispersive x-ray (EDX), x-ray diffraction (XRD), dynamic laser scattering (DLS), Brunauer–Emmett–Teller (BET) techniques and also magnetic characterization by vibrating sample magnetometer (VSM). The results show a correlation between the milling time and the crystallite and particle size, specific surface area, magnetic properties and nanoscale porosity of the iron powder. The iron powders obtained were a kind of mesoporous materials. The properties of the iron powder were examined with respect to their oxygen reducing kinetics.

Docynia indica (D. indica) shows various useful biological activities, such as antioxidant, anti-inflammatory, antibacterial effects, and positive benefits for human health. Such biological activities relate to the main phytochemicals of D. indica including phenolic and flavonoid. However, isolation for phenolic and flavonoid by popular methods such as hot extraction, soxhlet extraction, and ultrasonic extraction have been relatively ineffective. Therefore, in this study, microwave-assisted extraction (MAE) was used for the extraction of total phenolic and total flavonoid from D. indica. The optimization experiments were conducted based on response surface methodology (RMS) according to a central composite design with four independent variables: extraction time (min), ethanol concentration (%, v/v), microwave power (W), and pH of the solvent. Three dependent variables were total phenolic content (TPC), total flavonoid content (TFC), and yield. The optimal conditions for the extraction of phenolic and flavonoid from D. indica were: extraction time of 50 min, ethanol concentration of 65%, microwave power of 440 W, and solvent pH of 5.4. These conditions corresponded to TPC, TFC, and yield values of 33.57 ± 0.12 (mg GAE/g), 25.01 ± 0.11 (mg QE/g) and 33.44 ± 0.14 (%), respectively.

Rhizodegradation is a process by which plant-supplied substrates stimulate microbial populations in plant root zones (rhizospheres) to cause removal of undesirable levels of contaminants in soil. This study characterized rhizodegradation of the insecticide bifenthrin in Armour silt loam and Sullivan fine sandy loam soils that were planted with switchgrass, big bluestem, and alfalfa. After six weeks in soils, plate dilution frequency assays (PDFA) of bacterial populations were higher in all planted soils than in unplanted ones. Planted Sullivan soils contained higher bacteria than corresponding Armour soils and alfalfa rhizospheres of both soil types contained highest bacteria. Bacterial populations generally increased between week 6 and week 10, before declining in each treatment at week 12. Carbon utilization patterns (CUP) of bacterial communities, measured as color development on BIOLOG plates, were higher in planted soils than in unplanted ones. Principal Component Analysis (PCA) constructed patterns based on different extents of color development; these patterns were used to relate microbial communities in the different treatments. Gas chromatography (GC-ECD) showed that significantly more bifenthrin dissipated in planted soils than unplanted ones. Different levels of bifenthrin were recovered in planted soils but the differences were generally not significant. Data are being evaluated further to provide a basis for the development of strategies for enhancing rhizodegradation of soils contaminated with bifenthrin