Aquaporin 6 (AQP6) is an anion channel that is expressed primarily in acid secreting α-intercalated cells of the kidney collecting duct. In addition, AQP6 anion channel permeability is gated by low pH. Inspection of the N-terminus of AQP6 revealed a putative calmodulin binding site. AQP6-expressing CHO-K1 cell lysates were mixed with calmodulin beads and AQP6 was pulled down in the presence of calcium. Mutagenesis of the N-terminal calmodulin binding site in full length mouse AQP6 resulted in a loss of calmodulin binding activity. Mouse and human AQP6 calmodulin binding site peptides bound dansyl-calmodulin with a dissociation constant of approximately 1 μM. The binding of AQP6 to calmodulin may be an important key to determining the physiological role of AQP6 in the kidney.
Few analytical methods exist that combine chemical and sensory analysis of odorous compounds in whole air. Volatile organic compounds were collected by sampling air downwind from a small dairy through sorbent tubes of Tenax TA and Carboxen 569. Samples were analyzed by thermal desorption into a cryotrap and subsequent gas chromatographic separation, followed by simultaneous olfactometry and mass spectrometry. Because compounds are concentrated during sampling, sensory analysis encountered compounds at a concentration 40 times that in air, making this a useful method for identifying trace compounds participating in odor. Twenty odorous and nonodorous compounds were identified and quantified, including straight-chain and aromatic hydrocarbons, chlorinated compounds, alcohols, ketones, aldehydes, and organic acids, at air concentrations of 0.55-320.20 microg/m(3). Compound peaks were characterized by odors ranging from offensive to pleasant, demonstrating the integrative nature of olfaction. This method could be useful in studying many kinds of odors in air.
Few data are available on the ammonia emissions of large-scale outdoor animal facilities in arid climates such as those found in California's San Joaquin Valley. Passive samplers provide an ideal tool for studying such large and heterogeneous area sources, because they are inexpensive, portable, and fully self-contained. UC Davis passive ammonia samplers incorporate modifications on a previous design, the Willems Badge, for ease of analysis. Citric acid was chosen as a coating medium though it performed as well as oxalic, sulfuric, and tartaric acids. Zefluor PTFE prefilters were used instead of Teflo though both showed the same resistance to diffusion. Citric acid-coated filters were stable for up to 10 weeks, though more so if stored in Petri dishes rather than in the sampling cassettes themselves. The most effective sampler position was found to be in a face-down configuration fixed into the wind to avoid debris and sensitivity to wind shifts. A new method of rinsing the filters within the cassettes by dropwise elution proved highly effective, with 85% of the ammonium being removed in the first 3 mL of the 10-mL rinse volume. Application of the sampler at a dairy in the Joaquin Valley revealed large variations in concentrations at different locations along the downwind fenceline, which correlated with animal populations and activities directly upwind. In addition, large variations in ammonia concentrations were observed in relation to time of day and animal activity. Field blank loadings were of 1.40 microg NH4-N/filter (SD = 0.74 microg NH4-N/filter). Replicate passive samplers placed side-by-side during sampling episodes agreed with a slope of 1.010 (standard error = 0.028). Impingers were used as a reference method to obtain the correlation between filter loadings and air concentrations, yielding an "effective sampling rate" for the passive samplers of 6.18 L/h (error = 0.23 L/h). Using a theoretical calculation, that "effective flow rate" was calculated to be 6.29 L/h. The method's limit of detection was found to be 82.5 microg NH4-N/m3. Wind speed was found to theoretically affect linearity of sampler response only for speeds less than 0.92 m/s.
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