Imaging cytoplasmic cAMP in mouse brainstem neurons

Background  

cAMP is an ubiquitous second messenger mediating various neuronal functions, often as a consequence of increased intracellular Ca²⁺ levels. While imaging of calcium is commonly used in neuroscience applications, probing for cAMP levels has not yet been performed in living vertebrate neuronal tissue before.     

Detection and classification of gaseous sulfur compounds by solid electrolyte cyclic voltammetry of cermet sensor array

Electrochemical sensors composed of a ceramic-metallic (cermet) solid electrolyte are used for the detection of gaseous sulfur compounds SO₂, H₂S, and CS₂ in a study involving 11 toxic industrial chemical (TIC) compounds. The study examines a sensor array containing four cermet sensors varying in electrode-electrolyte composition, designed to offer selectivity for multiple compounds. The sensors are driven by cyclic voltammetry to produce a current-voltage profile for each analyte. Raw voltammograms are processed by background subtraction of clean air, and the four sensor signals are concatenated to form one vector of points. The high-resolution signal is compressed by wavelet transformation and a probabilistic neural network is used for classification. In this study, training data from one sensor array was used to formulate models which were validated with data from a second sensor array. Of the 11 gases studied, 3 that contained sulfur produced the strongest responses and were successfully analyzed when the remaining compounds were treated as interferents. Analytes were measured from 10 to 200% of their threshold-limited value (TLV) according to the 8-h time weighted average (TWA) exposure limits defined by the National Institute of Occupational Safety and Health (NIOSH). True positive classification rates of 93.3, 96.7, and 76.7% for SO₂, H₂S, and CS₂, respectively, were achieved for prediction of one sensor unit when a second sensor was used for modeling. True positive rates of 83.3, 90.0, and 90.0% for SO₂, H₂S, and CS₂, respectively, were achieved for the second sensor unit when the first sensor unit was used for modeling. Most of the misclassifications were for low concentration levels (such 10-25% TLV) in which case the compound was classified as clean air. Between the two sensors, the false positive rates were 2.2% or lower for the three sulfur compounds, 0.9% or lower for the interferents (eight remaining analytes), and 5.8% or lower for clean air. The cermet sensor arrays used in this analysis are rugged, low cost, reusable, and show promise for multiple compound detection at parts-per-million (ppm) levels.     

Stable isotopic studies of earthworm feeding ecology in tropical ecosystems of Puerto Rico

Feeding strategies of earthworms and their influence on soil processes are often inferred from morphological, behavioral and physiological traits. We used (13)C and (15)N natural abundance in earthworms, soils and plants to explore patterns of resource utilization by different species of earthworms in three tropical ecosystems in Puerto Rico. In a high altitude dwarf forest, native earthworms Trigaster longissimus and Estherella sp. showed less (15)N enrichment ((15)N = 3-6 per thousand) than exotic Pontoscolex corethrurus ((15)N =7-9 per thousand) indicating different food sources or stronger isotopic discrimination by the latter. Conversely, in a lower altitude tabonuco forest, Estherella sp. and P. corethrurus overlapped completely in (15)N enrichment ((15)N = 6-9 per thousand), suggesting the potential for interspecific competition for N resources. A tabonuco forest converted to pasture contained only P. corethrurus which were less enriched in (15)N than those in the forest sites, but more highly enriched in (13)C suggesting assimilation of C from the predominant C(4) grass. These results support the utility of stable isotopes to delineate resource partitioning and potential competitive interactions among earthworm species. Copyright 1999 John Wiley & Sons, Ltd.     

Chemiluminescence detection of hydrazine vapor

An efficient, real-time chemiluminescence detector for hydrazine vapor, N(2)H(4)(g), is described, capable of monitoring sub part-per-billion levels of hydrazine in air. The catalytic oxidation of hydrazine by colloidal platinum forms an intermediate, oxidizing agent (e.g. OH or OOH) which subsequently oxidizes luminol, generating a chemiluminescence signal that is proportional to the hydrazine concentration. Major components of the instrument include a photomultiplier tube (PMT), a short length of glass tubing coiled directly in front of the PMT cathode surface, a vacuum pump for sampling the air, and a peristaltic pump for circulating the liquid reagent. The liquid reagent, a basic solution (pH 13) of luminol and colloidal platinum, is continuously recycled. The detection sequence is initiated by pumping the hydrazine vapor through a short length of teflon tubing that is concurrently transporting the liquid reagent. The liquid is separated from the gas stream in an impinger and quickly pumped to the PMT. We have evaluated the effect of solution pH, luminol and platinum concentrations, and air and liquid flow rates on the analytical characteristics of this system. A linear, dynamic detection range for hydrazine has been obtained from 1 to 2000 ppb in air, with an instrument response that is fully reversible and achieves plateau response in less than 2 min.