Step Change In Concentration Research Articles

Reducing time and increasing sensitivity in sample preparation for adherent mammalian cell metabolomics.

A simple, fast, and reproducible sample preparation procedure was developed for relative quantification of metabolites in adherent mammalian cells using the clonal β-cell line INS-1 as a model sample. The method was developed by evaluating the effect of different sample preparation procedures on high performance liquid chromatography- mass spectrometry quantification of 27 metabolites involved in glycolysis and the tricarboxylic acid cycle on a directed basis as well as for all detectable chromatographic features on an undirected basis. We demonstrate that a rapid water rinse step prior to quenching of metabolism reduces components that suppress electrospray ionization thereby increasing signal for 26 of 27 targeted metabolites and increasing total number of detected features from 237 to 452 with no detectable change of metabolite content. A novel quenching technique is employed which involves addition of liquid nitrogen directly to the culture dish and allows for samples to be stored at -80 °C for at least 7 d before extraction. Separation of quenching and extraction steps provides the benefit of increased experimental convenience and sample stability while maintaining metabolite content similar to techniques that employ simultaneous quenching and extraction with cold organic solvent. The extraction solvent 9:1 methanol: chloroform was found to provide superior performance over acetonitrile, ethanol, and methanol with respect to metabolite recovery and extract stability. Maximal recovery was achieved using a single rapid (∼1 min) extraction step. The utility of this rapid preparation method (∼5 min) was demonstrated through precise metabolite measurements (11% average relative standard deviation without internal standards) associated with step changes in glucose concentration that evoke insulin secretion in the clonal β-cell line INS-1.

EFFECTS OF ROUGHNESS ON THE OXYGEN TRANSFER AT THE SEDIMENT WATER INTERFACE - IMPORTANCE OF NONSTEADY PHENOMENA AND BIOCHEMICAL PROCESSES -

A numerical model was developed to calculate diffusive dissolved-oxygen (DO) transfer rate at rough sediment surface, considering vortex shedding in the water and biochemical reactions in the sediment. Calculational results showed that non-dimensionalized DO transfer rate (Stanton number, St) had a maximum value in the transitional region of surface roughness, in which the DO flux was two-three times larger than that at the smooth surface. Calculational results was significantly modified by considering non-steady variations in diffusive transfer rate due to step changes in DO concentration in the flushed region. Quantitative estimation of DO concentration at the sediment-water interface was also a critical procedure to evaluate St values.

Effects of Hydrodynamic Conditions on DO Transfer at a Rough Sediment Surface

A numerical model was developed to calculate the rate of dissolved-oxygen (DO) diffusion across a sediment surface taking into account the surface roughness and biochemical reactions of the sediment. Estimates of DO transfer rate from the model were compared with results from laboratory experiments conducted in a rectangular flume using roughness elements. In experiments, there was maximum value for the nondimensionalized DO transfer rate (Stanton number, St) in the transitional region of surface roughness, in which the mass flux was two to five times larger than that of the smooth surface. The reproducibility of the experimental results by numerical analysis was significantly improved by including terms for flushing frequency of water in cavities between the roughness elements and for nonsteady variations in the diffusion rate due to step changes in DO concentration in the flushed region. A simple method to estimate enhancement effect for St caused by nonsteady variations was also presented.

PH transients in hydroxyapatite chromatography columns—Effects of operating conditions and media properties

pH transitions occur in hydroxyapatite (HAP) columns that are subject to step changes in salt concentration, which have been shown to be controlled by proton exchange on the HAP surface. The pH temporarily decreases before gradually returning to the feed value when the salt concentration increases, potentially compromising the stability of the HAP when either the magnitude or duration of the pH drop is excessive. The opposite happens when the salt concentration decreases. In this work we address the effects of several key variables: the flow rate, the particle size, the use of salt gradients instead of steps, the use of different co-buffers, the surface area of the HAP, and the use of a slightly alkaline wash prior to increasing the salt concentration. Flow rate and particle size were found to have virtually no effect, demonstrating that the pH transitions are equilibrium rather than kinetically driven. Salt gradients resulted in smaller pH drops compared to steps since the exchanged protons are diluted over the gradient volume. MES and histidine used as co-buffers were effective at reducing the duration of the pH transitions but did not affect their magnitude. The same result was found when comparing HAP samples with different surface areas, with the lower surface area HAP yielding much shorter duration but similar pH drops and rises. Finally, washing the HAP column with a pH 7.5 buffer prior to the salt step was found to dramatically reduce the subsequent pH drop. In general, there was good agreement between these results and predictions based on our previously developed model.

Effect of ammonium ion distribution on Nafion ® conductivity

Nafion ® conductivity in a proton exchange membrane fuel cell (PEMFC) with the fuel stream containing ammonia is mainly affected by the ammonium ion composition and operating conditions. In this study, the effect of ammonium ion distribution on Nafion conductivity was investigated for the first time. The conductivities of two kinds of contaminated membranes having uniform and non-uniform ammonium ion distributions were studied. To simulate a membrane with a well-defined ammonium ion concentration profile, three individual Nafion membranes containing known amounts of ammonium ions were physically stacked together. The uniform and non-uniform cases represented membranes having three layers with the same y N H 4 + or step changes in concentration, respectively. Under fuel cell operations, the conductivities of non-uniformly poisoned membranes were ca. 1.07–1.86 times larger than those of uniformly poisoned membranes, depending on humidity, contamination level, and ammonium ion distribution. Consequently, the performance prediction of a cationic-poisoned PEMFC needs to consider any concentration gradients that may exist in MEA. The liquid-phase conductivities of composite membranes were also studied and the results show that conductivity measurements performed in deionized water are not representative of what exists under fuel cell conditions due to rapid redistribution of ions in the Nafion via the liquid phase.

Bio-sensing using recessed gold-filled capillary amperometric electrodes

A novel recessed electrode is reported for amperometric detection of hydrogen peroxide and via glucose oxidase for the detection of glucose. The electrode utilised electrodeposited platinum over a gold wire surface, which proved to be an effective peroxide-detecting surface. Compared with a traditional exposed electrode surface, the recessed tip facilitated an extended linear range for glucose from 4 to over 14 mM. Bio-fouling, as assessed by exposure to bovine serum albumin, was also significantly reduced. Though response time at the recess was increased, it was within an acceptable range for physiological monitoring. Moreover, the recess enabled precise measurement of the hydrogen peroxide diffusion coefficient; this was based on a bipartite expression for the transient amperometric current at the recessed structure following a step change in ambient hydrogen peroxide concentration. An important aspect of the diffusion measurement was the curve fitting routine used to map on to the theoretical response curve.

Chemotactic Response and Adaptation Dynamics in Escherichia coli

Adaptation of the chemotaxis sensory pathway of the bacterium Escherichia coli is integral for detecting chemicals over a wide range of background concentrations, ultimately allowing cells to swim towards sources of attractant and away from repellents. Its biochemical mechanism based on methylation and demethylation of chemoreceptors has long been known. Despite the importance of adaptation for cell memory and behavior, the dynamics of adaptation are difficult to reconcile with current models of precise adaptation. Here, we follow time courses of signaling in response to concentration step changes of attractant using in vivo fluorescence resonance energy transfer measurements. Specifically, we use a condensed representation of adaptation time courses for efficient evaluation of different adaptation models. To quantitatively explain the data, we finally develop a dynamic model for signaling and adaptation based on the attractant flow in the experiment, signaling by cooperative receptor complexes, and multiple layers of feedback regulation for adaptation. We experimentally confirm the predicted effects of changing the enzyme-expression level and bypassing the negative feedback for demethylation. Our data analysis suggests significant imprecision in adaptation for large additions. Furthermore, our model predicts highly regulated, ultrafast adaptation in response to removal of attractant, which may be useful for fast reorientation of the cell and noise reduction in adaptation.

Proton exchange membrane fuel cell contamination model: Competitive adsorption followed by a surface segregated electrochemical reaction leading to an irreversibly adsorbed product

A generic, transient fuel cell kinetic loss mathematical model was developed for the case of contaminants that partially cover the catalyst surface with irreversibly adsorbed products. The model was derived using step changes in contaminant concentration, constant operating conditions and disregarding liquid water scavenging effects. The closed form solutions were validated using H 2S, SO 2 and COS data from a single source. The model needs to be validated against other data sets and transient operating conditions more representative of automotive applications. A method is proposed to determine kinetic rate constants and relies on tests with a reactant, a contaminant and, a reactant and a contaminant mixture. The method is useful to evaluate the presence of interactions between reactant and contaminant related adsorbates, and, to minimize electrode potential variations during controlled cell voltage measurements. Model parameters were similar for all contaminants suggesting a common adsorbate configuration. The model also expands the number of previously derived cases. All models in this inventory, derived with the assumption that the reactant is absent, lead to different dimensionless current vs. time behaviors similar to a fingerprint. These model characteristics facilitate contaminant mechanism identification. Model predictions include a limit of 0.7 ppb contaminant concentration in the reactant stream to minimize cell performance losses during the 5000 h automotive application life. This tolerance limit represents a worse case scenario because it does not take into account performance recovery resulting from drive cycle operation or the addition of mitigation strategies. A cell performance loss increase of 40% is also predicted for a catalyst loading decrease from 0.4 to 0.1 mg Pt cm −2.

A fixed-grid numerical modelling of transient liquid phase bonding and other diffusion-controlled phase changes

A transient liquid phase (TLP), in which a liquid layer is formed and subsequently solidifies, and other diffusion-controlled phase changes are generally associated with moving phase-change interfaces. Both fixed and variable grid discretization models have been formulated to investigate these diffusion-controlled problems. However, all numerical efforts to date have employed one of the approaches explicitly to track the moving interfaces across which there exist step changes in concentrations. In this article, the fixed-grid source-based method originally developed to simulate the temperature fields for melting-solidification phase change processes has been adopted to simulate diffusion-controlled dissolution and solidification. This method solves a unique diffusion equation for the different phases and the moving interfaces using implicit time integration. Compared with previously developed models, it is not only simpler in numerical formulation and procedure, but also more convenient to extend to many phases and high-dimensional problems. We report here the detailed formulation of the relevant equations, and compare and validate the model using experimental data and previous modelling predictions for several systems available from the existing literature.

Mathematical Model of Basal and Agonist-Dependent GIRK Channel Activity

We developed a model of GIRK channel activity. The channel activation scheme was based on sequential non-cooperative binding of 4 Gβγ molecules to channel protein (generating 5 closed states) and Gβγ independent channel opening. The kinetics of Gβγ interaction with subsequent change of channel conformation were adjusted to generate activation time of ∼ 1 s for a step rise in Gβγ concentration. The kinetics of switch from closed to open conformation were derived from single-channel analysis of GIRK1/2 recordings in Xenopus leaves oocytes. For simulation of agonist-dependent channel activation we incorporated the above scheme into a general model of G-protein cycle. This model was derived from that of Thomsen-Jaquez-Neubig. Several features were added: a) receptor was allowed to couple to G-protein in agonist-bound and in free state; b) finite affinity of Gα to Gβγ was assumed in GTP- and GDP-bound states; c) microscopic reversibility was obeyed in cyclic schemes containing reversible reactions; d) the assumption that G-protein concentration exceeded the receptor concentration was relaxed in order to enable simulation of titration experiments. We simulated the time-course of channel activation induced by step change in agonist concentration in presence and in absence of Gβγ-scavenging protein. We also simulated receptor-titration experiments. The results of simulations were compared to whole-cell experiments in Xenopus leaves oocytes. Our model produced realistic time course of channel activation and also demonstrated decremental dependence of activation time on receptor concentration. Comparing the simulation results with those expected from binary shuttle model of channel activation based on considerations of free diffusion of membrane proteins lead to the conclusion that G-protein activation by receptor is probably of catalytic collision-coupling type, while the channel and G protein were either in a tight complex or diffused in a restricted membrane domain.

Effect of copper loading on copper-ceria catalysts performance in CO selective oxidation for fuel cell applications

Copper–ceria catalysts with three different Cu loadings (1, 7 and 15 wt%) were prepared by incipient wet impregnation, dried at 120 °C and calcined in air at 500 °C. The as-prepared catalysts were characterized by XRD, BET, Diffuse Reflectance Spectroscopy (DRS–UV–visible), Raman spectroscopy, CO and H 2-TPR, CO-TPR, CO-TPD and Oxygen Storage Capacity (OSC) measurements (with CO and O 2 concentration step-changes). The results indicated a good dispersion of copper for catalysts with 1 and 7 wt% Cu; however, bulk CuO was present for catalyst with 15 wt% Cu loading. Catalyst with 7 wt% Cu was observed to have very high capacity to release lattice oxygen to oxidize CO at low temperature. Activity results for CO oxidation in the absence and in the presence of 60% H 2, demonstrated a very similar performance for catalysts with 7 and 15 wt% Cu (both with T 100 = 112 °C), and much better than that of catalyst loaded with 1 wt% Cu. Catalyst with 7 wt% of copper shows very high activity (100% in a wide temperature window) and selectivity (higher than 85%), which makes an attractive for its use in purification of hydrogen for fuel cell applications. The presence of a mixture of CO 2 and H 2O inhibited catalyst activity, with CuO/CeO 2 catalyst with 7 wt% Cu exhibiting the best performance in the overall reaction temperature range. This could be attributed to the presence of highly disperse copper, only part of it in deep interaction with ceria. The effect of O 2/CO ratio ( λ) and the potential reversibility of the inhibitory effect of CO 2 and H 2O were also investigated.

Modification of a commercial cavity ring-down spectroscopy NO2 detector for enhanced sensitivity

Nitrogen dioxide (NO(2)) plays a central role in atmospheric chemistry, air pollution, and biogeochemical cycles. Many analytical techniques have been developed to detect NO(2), but only chemiluminescence-based instruments are commonly, commercially available. There remains a need for a fast, light, and simple method to directly measure NO(2). In this work we describe the modification and characterization of a small, commercially available cavity ring-down spectroscopy (CRDS) NO(2) detector suitable for surface and aircraft monitoring. A metal oxide scrubber was added to remove NO(2), and provide a chemical zero, improving the detection limit (3sigma of the background noise) from several parts per billion by volume (ppbv) to 0.06 ppbv, integrated over 60 s. Known interferences by water and particles were removed using Nafion tubing and a 1 microm Teflon filter, respectively. A 95% response time of 18+/-1 s was observed for a step change in concentration. The CRDS detector was run in parallel to an ozone chemiluminescence device with photolytic conversion of NO(2) to NO. The two instruments measured ambient air in suburban Maryland. A least-squares fit to the comparison data resulted a slope of 0.960+/-0.002 and R of 0.995, showing agreement within experimental uncertainty.

Improved response time with a new miniaturised main-stream multigas monitor

For paediatric monitoring and demanding applications such as metabolic monitoring and measurements of functional residual capacity combining gas concentration with flow/volume measurements the performance of side-stream monitors (SSGM) is suboptimal. The objective was to evaluate the performance of a miniaturised mainstream multigas monitor (MSGM) alleged to offer fast response gas monitoring. The MSGM uses infrared technique for measurements of carbon dioxide, nitrous oxide and inhalation agents and fuel cell technique for oxygen monitoring. The MSGM performance was com- pared to a state of the art side-stream monitor in a bench study. Response time was measured in two bench study set ups; a high flow oxygen flush to achieve one step change in gas concentrations and during continuous ventilation using a circuit with an oxygen consuming/carbon dioxide producing lung model connected to a ventilator. Averaged tracings from the tested monitors were used for calculation of the 90-10% decline of CO(2), the corresponding 10-90% incline of O(2) and N(2)O and of Isoflurane concentrations in the flush set up and at different inspired O(2) for the O(2) upslope and corresponding CO(2) down- slope during continuous ventilation at different breathing frequencies. Calibration gases with different concentrations of CO(2), O(2) and N(2)O were used for testing of accuracy. The MSGM response time for CO(2) was 96 (88-100) compared to 348 (340-352) ms for the SSGM (P < 0.001). Corresponding response times for O(2) was 108 (76-144), and 432 (360-448) ms (P < 0.001), respectively. At a respiratory rate of 60 BPM the SSGM trace was damped and sinusoidal whereas the MSGM displayed wider amplitude and a square waveform. The deviations from calibration gas values were within clinically acceptable range and linear for all gases over the concentration range studied for both monitors. The MSGM response time for CO(2) and O(2) was less than 1/3 of the SSGM. The performance of the MSGM was maintained at high breathing frequencies. The accuracy was within clinically acceptable limits for both monitors.

Transient modeling of anode-supported solid oxide fuel cells

An isothermal 2-D transient model is developed for an anode-supported solid oxide fuel cell. The model takes into account the transient effects of both charge migration and species transport in PEN assembly. Due to the lack of transient experimental data, the transient model, under steady state operating conditions, is validated using experimental results from open literature. Numerical results show that the cell can obtain very quick transient current response when subjected to a step voltage change, followed by a slow current transient period due to species diffusion effects within porous electrodes. It is also found that the transient response of the cell current is sensitive to oxygen concentration change at cathode/channel interface, whereas the current response is slow when step change of hydrogen concentration is applied at anode/channel interface. The cell transient performance can be improved by increasing porosity or decreasing tortuosity of electrodes.

Avian Chemoreception

The first detailed physiological evidence for olfactory and trigeminal chemoreception in an avian species is provided by a series of investigations in the chicken (Gallus domesticus). Initial work indicated that the activity of avian olfactory bulb neurones closely resembles that of other vertebrates, exhibiting variable spontaneous temporal firing patterns with mean firing rates between those reported for mammals and reptiles. Application of odors directly to the olfactory epithelium showed that like mammals, avian olfactory bulb neurones respond in the form of inhibition and excitation with accompanying changes in temporal firing pattern. When exposed to a range of concentrations of a single odor, all responsive neurones exhibited an ability to discriminate small step-changes in concentration producing clear stimulus response relationships. Avian trigeminal chemoreception was also investigated by examining the responses of single mucosal receptors in the nasal cavity and palate. Slowly and rapidly adapting nasal mechanoreceptors were identified, some of which exhibited chemical sensitivity when exposed to ammonia gas, acetic acid vapor or carbon dioxide. These results demonstrate that polymodal nociceptors are present in avian nasal mucosa and represent the first attempt in any species to quantify the responses of single trigeminal receptors to a range of concentrations of noxious airborne chemicals. Collectively, the findings demonstrate how an electrophysiological approach can improve our understanding of the underlying sensory physiology relating to avian perception of the chemical environment.

Dynamic Characteristics of a Fast-Response Aerosol Size Spectrometer

The fast integrated mobility spectrometer (FIMS) is a highly sensitive instrument with a fast response time. The time response of the FIMS is limited by the mixing of the aerosol in the inlet of the instrument, which “smears” the detected aerosol over a range of time. The response time is also limited by the different transit times that particles experience in the instrument due to differences in particle classification trajectories. Experiments show that the difference in particle transit times can be corrected using a simple model of the particle trajectories. Furthermore, the “smearing” effects in the inlet can be corrected by de-convolving the time series of particle counts (the temporal de-convolution) in each size bin before inverting the data. The dynamic response of the FIMS was investigated by measuring an aerosol subjected to a step-change and sinusoidal-change in number concentration. The attenuation of the FIMS signal, without using the temporal de-convolution, was measured with and without an aerosol neutralizer in the FIMS inlet. Due to its large internal volume, the neutralizer significantly slows down the time response of the FIMS when installed in the inlet. For a sinusoidal signal at 0.33 Hz, the measured attenuation of the FIMS without the neutralizer was 56% versus an attenuation of 82% when the neutralizer was used. When the temporal de-convolution was applied at the same frequency, the FIMS was able to capture the full variation of the aerosol size distribution; however the random noise in the derived size distribution was amplified.

Textural characterization of media composed of compacted pieces of cardboard and polyethylene using a gas tracer method

The aim of this work is the experimental determination of effective transport properties of porous media consisting of compacted pieces of cardboard and polyethylene (PE). The proposed method itself is more general and can be applied to many different materials and contexts. Three major transport properties were determined: porosity, tortuosity factor and permeability. Three parameters characterizing the media were varied over a wide range: the bulk density, the size of the elements entering the mix, and the proportion of cardboard and PE in the mix. The properties were measured by means of a specially designed experimental device based on miscible gas tracing. The porosity and tortuosity factor were simultaneously determined by parametric identification, based on the experimental sample output response to an inlet gas concentration step change compared to the results of a direct numerical model. Permeability was calculated in the standard way from the measurement of the pressure drop across the sample. The reproducibility of the measurements was very good. It was found that changing the material density of the medium significantly affects all three structural properties. When the bulk density is varied between 300 and 900 kg m(-3), the tortuosity factor varies in a range as large as 18-8 and the permeability decreases by a ratio of 2-3. The tortuosity factor shows unusual variation, characterized by a decrease when density is increased above 500 kg m(-3). The size of the elements does not significantly affect the structural properties of the medium in the range of parameters studied.

Fabrication and Optimization of a Nanoporous Platinum Electrode and a Non-enzymatic Glucose Micro-sensor on Silicon

In this paper, optimal conditions for fabrication of nanoporous platinum (Pt) were investigated in order to use it as a sensitive sensing electrode for silicon CMOS integrable non-enzymatic glucose micro-sensor applications. Applied charges, voltages, and temperatures were varied during the electroplating of Pt into the formed nonionic surfactant C16EO8 nano-scaled molds in order to fabricate nanoporous Pt electrodes with large surface roughness factor (RF), uniformity, and reproducibility. The fabricated nanoporous Pt electrodes were characterized using atomic force microscopy (AFM) and electrochemical cyclic voltammograms. Optimal electroplating conditions were determined to be an applied charge of 35 mC/mm2, a voltage of -0.12 V, and a temperature of 25 °C, respectively. The optimized nanoporous Pt electrode had an electrochemical RF of 375 and excellent reproducibility. The optimized nanoporous Pt electrode was applied to fabricate non-enzymatic glucose micro-sensor with three electrode systems. The fabricated sensor had a size of 3 mm × 3 mm, air gap of 10 μm, working electrode (WE) area of 4.4 mm2, and sensitivity of 37.5 μA•L/mmol•cm2. In addition, it showed large detection range from 0.05 to 30 mmolL-1 and stable recovery responsive to the step changes in glucose concentration.

Fabrication and Optimization of a Nanoporous Platinum Electrode and a Non-enzymatic Glucose Micro-sensor on Silicon.

In this paper, optimal conditions for fabrication of nanoporous platinum (Pt) were investigated in order to use it as a sensitive sensing electrode for silicon CMOS integrable non-enzymatic glucose micro-sensor applications. Applied charges, voltages, and temperatures were varied during the electroplating of Pt into the formed nonionic surfactant C16EO8 nano-scaled molds in order to fabricate nanoporous Pt electrodes with large surface roughness factor (RF), uniformity, and reproducibility. The fabricated nanoporous Pt electrodes were characterized using atomic force microscopy (AFM) and electrochemical cyclic voltammograms. Optimal electroplating conditions were determined to be an applied charge of 35 mC/mm2, a voltage of -0.12 V, and a temperature of 25 °C, respectively. The optimized nanoporous Pt electrode had an electrochemical RF of 375 and excellent reproducibility. The optimized nanoporous Pt electrode was applied to fabricate non-enzymatic glucose micro-sensor with three electrode systems. The fabricated sensor had a size of 3 mm x 3 mm, air gap of 10 µm, working electrode (WE) area of 4.4 mm2, and sensitivity of 37.5 µA•L/mmol•cm2. In addition, it showed large detection range from 0.05 to 30 mmolL-1 and stable recovery responsive to the step changes in glucose concentration.

Mechanical Ventilator for Delivery of 17O2 in Brief Pulses

The 17O nucleus has been used recently by several groups for magnetic resonance (MR) imaging of cerebral metabolism. Inhalational delivery of 17O2 in very brief pulses could, in theory, have significant advantages for determination of the cerebral metabolic rate for oxygen (CMRO2) with MR imaging. Mechanical ventilators, however, are not typically capable of creating step changes in gas concentration at the airway. We designed a ventilator for large animal and human studies that provides mechanical ventilation to a subject inside an MR scanner through 25 feet of small-bore connecting tubing, and tested its capabilities using helium as a surrogate for 17O2. After switching the source gas from oxygen to helium, the 0-90% response time for helium concentration changes at the airway was 2.4 seconds. The capability for creating rapid step changes in gas concentration at the airway in large animal and human studies should facilitate the experimental testing of the delivery 17O2 in brief pulses, and its potential use in imaging CMRO2.