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The mechanical basis of Drosophila audition.

In Drosophila melanogaster, antennal hearing organs mediate the detection of conspecific songs. Combining laser Doppler vibrometry, acoustic near-field measurements and anatomical analysis, we have investigated the first steps in Drosophila audition, i.e. the conversion of acoustic energy into mechanical vibrations and the subsequent transmission of vibrations to the auditory receptors in the base of the antenna. Examination of the mechanical responses of the antennal structures established that the distal antennal parts (the funiculus and the arista) together constitute a mechanical entity, the sound receiver. Unconventionally, this receiver is asymmetric, resulting in an unusual, rotatory pattern of vibration; in the presence of sound, the arista and the funiculus together rotate about the longitudinal axis of the latter. According to the mechanical response characteristics, the antennal receiver represents a moderately damped simple harmonic oscillator. The receiver's resonance frequency increases continuously with the stimulus intensity, demonstrating the presence of a non-linear stiffness that may be introduced by the auditory sense organ. This surprising, non-linear effect is relevant for close-range acoustic communication in Drosophila; by improving antennal sensitivity at low song intensities and reducing sensitivity when intensity is high, it brings about dynamic range compression in the fly's auditory system.

Acoustic Stimulation↗

Analysis of gaseous toxic industrial compounds and chemical warfare agent simulants by atmospheric pressure ionization mass spectrometry.

The suitability of atmospheric pressure chemical ionization mass spectrometry as sensing instrumentation for the real-time monitoring of low levels of toxic compounds is assessed, especially with respect to public safety applications. Gaseous samples of nine toxic industrial compounds, NH3, H2S, Cl2, CS2, SO2, C2H4O, HBr, C6H6 and AsH3, and two chemical warfare agent simulants, dimethyl methylphosphonate (DMMP) and methyl salicylate (MeS), were studied. API-MS proves highly suited to this application, with speedy analysis times (<30 seconds), high sensitivity, high selectivity towards analytes, good precision, dynamic range and accuracy. Tandem MS methods were implemented in selected cases for improved selectivity, sensitivity, and limits of detection. Limits of detection in the parts-per-billion and parts-per-trillion range were achieved for this set of analytes. In all cases detection limits were well below the compounds' permissible exposure limits (PELs), even in the presence of added complex mixtures of alkanes. Linear responses, up to several orders of magnitude, were obtained over the concentration ranges studied (sub-ppb to ppm), with relative standard deviations less than 3%, regardless of the presence of alkane interferents. Receiver operating characteristic (ROC) curves are presented to show the performance trade-off between sensitivity, probability of correct detection, and false positive rate. A dynamic sample preparation system for the production of gas phase analyte concentrations ranging from 100 pptr to 100 ppm and capable of admixing gaseous matrix compounds and control of relative humidity and temperature is also described.

Air Pollutants↗

Middle latency responses to electrical stimulation of the auditory nerve in unanaesthetized guinea pigs.

Middle latency responses (MLR) to sinusoidal and pulsatile electrical stimulation (ES) of the cochlea and to acoustical stimulation (AS) were evaluated in awake guinea pigs with chronically implanted electrodes. The ear, which was later electrically stimulated, was deafened by local intracochlear application of gentamicin, the opposite ear was left intact. Waveforms and P1-P2 interpeak intervals of the electrically evoked MLR (ES-MLR) were similar to those evoked by acoustical stimulation of the intact ear (AS-MLR) and the latencies of the ES-MLR were shorter by about 1-3 ms. Thresholds of ES-MLR in the frequency range 0.5-32 kHz increased with increasing ES frequency (slope 3.2 dB/octave), thresholds were 3.5-9.5 dB lower for intracochlear than for extracochlear ES. Dynamic ranges for ES-MLR varied between 6-20 dB. MLR amplitude-intensity functions for ES were steeper (slope 2-12 microV/dB) than those for AS (slope 0.2-2 microV/dB). Maximal ES-MLR amplitudes exceeded usually 1.5-3 times the amplitudes of the acoustically evoked MLR. Both types of stimulations evoked larger MLR amplitudes to contralateral stimulation than to ipsilateral stimulation (average ratio = 4.1 +/- 2.2 for AS and 3.3 +/- 2.2 for ES). Because of the relatively long latency and therefore insensitivity to electrical artifact, the ES-MLR can be used for the evaluation of different strategies of the electrical stimulation of the cochlea in awake guinea pig.

Acoustic Stimulation↗

Analysis of small-molecule interactions using Biacore S51 technology.

Biacore S51 is a new surface plasmon resonance-based biosensor developed by Biacore AB (Uppsala, Sweden). The instrument was engineered specifically to support small-molecule drug discovery and development. The platform includes increased sensitivity, larger sample handling capabilities, and automated data processing to improve throughput. Compared to previously released Biacore instruments, the most significant design change relates to the introduction of the hydrodynamic-addressing flow cell. This design allows two reaction surfaces and a reference surface to be placed within the same flow cell, thereby improving data quality and extending the kinetic range of the instrument. Using a set of small-molecule inhibitors of the enzyme carbonic anhydrase II, we tested the reproducibility, sensitivity, and dynamic range of the biosensor. Given the S51's performance capabilities, it should play an active role in secondary screening by providing high-resolution information for small-molecule/target interactions.

Carbonic Anhydrase II↗

A screening method for antigen-specific IgE using mast cells based on intracellular calcium signaling.

A simple screening method is presented for the measurement of antigen-specific IgEs in sera in which mast cells are used. This method is based on the intracellular calcium signal in mast cells induced by cross-linking the surface high-affinity Fc receptors (FcepsilonRIs) with IgEs and multivalent antigens. When a serum containing various antigen-specific IgEs is added to the mast cell suspension, various antigen-specific IgEs are captured by FcepsilonRIs on the cell surface. However, the required antigen-specific IgE can be specifically detected after the addition of the corresponding antigen. The resulting increase in intracellular calcium concentration ([Ca2+]i), monitored by Ca2+-fluorometry, was found to be an analytical measure for the screening of IgEs. Two kinds of rodent mast cells, cell-lined RBL2H3 cells and primary cultured BMMCs, were used as a representative model system of mast cells. A DNP hapten (DNP35-HSA) and ovalbumin (OVA) were chosen for illustrative antigens, and these antigen-specific IgEs (DNP-specific IgE, OVA-specific IgE) in the corresponding rodent sera were target antibodies. It was found that [Ca2+]i increased linearly with IgE concentrations ranging from 25 to 5000 ng/mL for DNP-specific IgE and from 5 to 50 ng/mL for OVA-specific IgE. For these dynamic ranges, optimum concentrations of antigens were found to be 10 ng/mL and 1 microg/mL for DNP35-HSA and OVA, respectively. It was concluded that by monitoring the increase of [Ca2+]i in mast cells, we could determine the antigen-specific IgEs. The present immunological assay based on the Ca2+ signal transduction in mast cells offers new possibilities for efficient screening of antigen-specific IgEs and the immunogenicity of IgE in sera.

Animals↗

Open multipass absorption cell for in situ monitoring of stratospheric trace gas with telecommunication laser diodes.

A two-mirror multipass absorption cell that is operated open to the atmosphere from a stratospheric balloon to monitor in situ methane (in the 1.65-microm region) and water vapor (in the 1.39-microm region) with telecommunication laser diodes is described. A small Cassegrain-type telescope is used to couple the cell simultaneously with two near-infrared InGaAsP laser diodes by means of optical fibers. The 1-m cell provides an absorption path length of 56 m. The optical cell was carefully designed to be free of incidental fringing in the 10(-5) absorption range. It is used in combination with a dual-beam detector to obtain a detection limit of 10(-5) absorption units, a large dynamic range of the measurements of many orders of magnitude, and a precision error in the concentration determination of a few percents. The optical arrangement of the cell and its ability to be used to detect in situ trace gas in the stratosphere, in severe environmental conditions, are exposed.

Journal Article↗

Fiber loop ringdown for physical sensor development: pressure sensor.

A new method of developing optical fiber pressure sensors by use of a fiber loop ringdown scheme is described. The fiber loop ringdown system is characterized in terms of the ringdown baseline stability, fiber transmission loss, and fiber refractive index. The overall sensor performance is demonstrated by use of sensing forces applied to the sensor head. The current device can sense pressures in the range of 0 to 9.8 x 10(6) Pa, converted approximately from the applied forces. The sensor's linear response, repeatability, detection sensitivity, measuring dynamic range, and temperature tolerance are explored.

Journal Article↗

Pressure transmission across the respiratory system at raised lung volumes in infants.

Forced expiratory flow-volume (FEFV) curves can be generated from end-tidal inspiration in infants with use of an inflatable jacket. We have developed a technique to raise lung volume in the infant before generation of FEFV curves. Measurements of pressure transmission to the airway opening by use of static maneuvers have shown no change with increasing lung volume above end-tidal inspiration. The aim of this study was to determine, under dynamic conditions (i.e., during rapid thoracic compression), whether the efficiency of pressure transmission across the chest wall is altered by raising lung volume above the tidal range. Dynamic pressure transmission (Ptx,dyn) was measured in five infants (age 6-17 mo). Jacket pressure (Pj), esophageal pressure, and volume were measured throughout passive and FEFV curves at lung volumes set by 10, 15, and 20 cmH2O preset pressure. The group mean Ptx,dyn was 37 +/- 6% (SE) of Pj at end-tidal inspiration, and no change was seen with further increases in lung volume. However, a mean decrease in Ptx,dyn of 42% was evident throughout the tidal volume range (i.e., from end-tidal inspiration to end expiration). Isovolume static pressure transmission (Ptx,st) was measured in three of the five infants by inflation of the jacket in a stepwise manner with the airway closed. Measurements were made at end-tidal inspiration and lung volumes at 10, 15, and 20 cmH2O preset pressure. Resulting changes in Pj, esophageal pressure, and airway opening pressure were compared using linear regressions to determine Ptx,st.(ABSTRACT TRUNCATED AT 250 WORDS)

Cystic Fibrosis↗

Antennal sucrose perception in the honey bee (Apis mellifera L.): behaviour and electrophysiology.

Physiological mechanisms of antennal sucrose perception in the honey bee were analysed using behavioural and electrophysiological methods. Following sucrose stimulation of the tip of a freely moving antenna, the latency of proboscis extension was 320-340 ms, 80-100 ms after the first activity in muscle M17 controlling this response. When bees were allowed to actively touch a sucrose droplet with one antenna, contacts with the solution were frequent with durations of 10-20 ms and average intervals between contacts of approximately 40 ms. High sucrose concentrations led to short and frequent contacts. The proboscis response and M17 activity were largely independent of stimulus duration and temporal pattern. Taste hairs of the antennal tip displayed spike responses to sucrose concentrations down to at least 0.1%. The first 25 ms of the response were suitable for discrimination of sucrose concentrations. This time interval corresponds to the duration of naturally occurring gustatory stimuli. Sucrose responses between different hairs on the same antenna showed a high degree of variability, ranging from less than five to over 40 spikes per 0.5 s for a stimulus of 0.1% sucrose. This variability of receptor responses extends the dynamic range of sucrose perception over a large range of concentrations.

Action Potentials↗

Ontogeny of neural discharge patterns in the ventral cochlear nucleus of the mongolian gerbil.

Discharge patterns were recorded extracellularly from single neurons in the ventral cochlear nucleus (VCN) of Mongolian gerbils ranging in age from 10 days after birth (DAB) to adult, a period which includes the onset of responsiveness to acoustic stimulation. At 10 DAB none of the neurons encountered within the VCN responded to acoustic stimulation. At 12 DAB approximately 15% of the neurons isolated in VCN were responsive. This coincided with the earliest cochlear microphonic potentials and preceded the appearance of the cochlear compound action potential (AP) by two days. At 14 DAB, or older, the great majority of neurons isolated in VCN responded to acoustic stimulation. Most parameters of VCN neural function exhibited significant changes between 12 and 18 DAB: neural thresholds improved approximately 100 dB; mean spontaneous discharge rate increased; the high-frequency range of characteristic frequency (CF) values increased from 10.0 to 24.0 kHz; the upper limit for phase locking increased from 0.8 kHz to 3.0 kHz; dynamic range increased from 16 dB to 44 dB, and the proportion of units with well-defined initial onset peaks in their post-stimulus-time (PST) response patterns increased from 40% to 100% of units. Most of the neural parameters examined achieved adult characteristics by 18 DAB. Frequency tuning (Q10dB) matured earlier for high-CF units. The most sharply tuned neurons with high CFs (greater than 4 kHz) at 12 DAB had Q10dB values equal to those for adults. None of the neurons with low CFs (less than 4 kHz) had Q10dB values greater than 1.2 at this age. Classical 'on' PST response patterns were not seen at 12 and 14 DAB. A unique PST response type, characterized by very long latency phasic discharge, was observed only at 12 DAB. None of the VCN neurons recorded from 12 DAB subjects displayed rhythmic 'bursting' or 'pulsing' PST response patterns, as has been reported at the earliest stages of functional development in the VCN of the cat. Most units were capable of sustained discharge, even with long stimulus durations. Units with 'primary-like' PST response patterns at 12 exhibited greater variability in first spike latency and less pronounced initial rates of firing than was characteristic in adults, resulting in poorly defined onset peaks. In contrast, units with chopper PST response patterns showed well-defined onset peaks.

Animals↗

Single cochlear fibre responses in guinea pigs with long-term endolymphatic hydrops.

Some cochlear fibre response properties have been measured in two GPs approximately one year after induction of endolymphatic hydrops (by surgical obliteration of the endolymphatic sac and duct). These animals are considered as models of the effects of hydrops in Menière's disease, and the purpose of the study was to examine any modifications of fibre response properties which may underly auditory symptoms of the disease in man. Neurones towards more apical cochlear regions (with low characteristic frequencies) showed the greatest deterioration in tuning properties; on average, in the 1-6 kHz range, Q10dB values were reduced by a factor of two compared with normal animals. Discharge rate versus intensity functions of such units were abnormally steep, with dynamic ranges reduced by 10-20 dB. Towards higher frequency regions neurone response properties showed less deterioration (contrasting with many other types of cochlear pathology where, in general, the high frequency basal region exhibits greatest vulnerability). We have also observed in a few units an abnormal bursting in both spontaneous and driven discharge. Interspike intervals during burst are less than 1 ms (within relative refractory period). These findings are related to the auditory symptoms of Menière's disease, in particular, poor frequency selectivity, loudness recruitment and tinnitus.

Action Potentials↗

A microcosm of musical expression: II. Quantitative analysis of pianists' dynamics in the initial measures of Chopin's Etude in E major.

Patterns of expressive dynamics were measured in bars 1-5 of 115 commercially recorded performances of Chopin's Etude in E major, op. 10, No. 3. The grand average pattern (or dynamic profile) was representative of many performances and highly similar to the average dynamic profile of a group of advanced student performances, which suggests a widely shared central norm of expressive dynamics. The individual dynamic profiles were subjected to principal components analysis, which yielded Varimax-rotated components, each representing a different, nonstandard dynamic profile associated with a small subset of performances. Most performances had dynamic patterns resembling a mixture of several components, and no clustering of of performances into distinct groups was apparent. Some weak relationships of dynamic profiles with sociocultural variables were found, most notably a tendency of female pianists to exhibit a greater dynamic range in the melody. Within the melody, there were no significant relationships between expressive timing [Repp, J. Acoust. Soc. Am. 104, 1085-1100 (1998)] and expressive dynamics. These two important dimensions seemed to be controlled independently at this local level and thus offer the artist many degrees of freedom in giving a melody expressive shape.

Female↗

Sonic spray ionization applied to liquid chromatography/mass spectrometry analysis of endocrine-disrupting chemicals in environmental water samples.

A sonic spray ionization liquid chromatography/mass spectrometry (LC/SSI-MS) procedure combined with off-line solid-phase extraction was optimized for the analysis of 20 endocrine-disrupting chemicals (EDCs) in water samples. Method development included a comparison of the novel sonic spray ionization (SSI) with more traditional ion sources, i.e. pneumatically assisted electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI). It was demonstrated that SSI and ESI spectra were very similar, but were more prone to the formation of solvent cluster ions as compared with APCI spectra. This phenomenon was most prominent for SSI and resulted in an increased chemical background in full-scan mass spectra. However, this chemical noise did not affect the overall sensitivity of SSI and ESI. After optimization of LC and MS parameters, the LC/SSI-MS method was validated. Recoveries ranged from 76.3 up to 113.4% for all compounds. Limits of detection (LOD) and quantitation (LOQ) were established between 3.0 and 11.5 ng/L and 9.9 and 38.0 ng/L, respectively. Within-day (n = 5) and between-day (n = 5) reproducibility were investigated at three levels and ranged from 3.3-16.5% and 7.6-19.2%, respectively. Eight-point calibration curves were established and showed linearity for all compounds (r(2) > 0.987) over a linear dynamic range of 10-10 000 ng/L.

Chromatography, Liquid↗

Liquid chromatographic separation of phenolic drugs using catalytic detection: comparison of an enzyme reactor and enzyme electrode.

A catalytic detection system based on the use of immobilized tyrosinase and two different transducers (photometric and electrochemical) is described. Comparison between tyrosinase immobilized in a packed-bed reactor and at the surface of a graphite electrode is discussed in terms of sensitivity in a flow injection system. The enzyme electrode configuration gives the highest sensitivity for the quantitation of dopamine. For the immobilized tyrosinase reactor with photometric detection the range for dopamine is linear up to 0.75 mM (136 micrograms ml-1) and the immobilized tyrosinase reactor with electrochemical detection and the tyrosinase electrode extends this dynamic range to 1 mM (181 micrograms ml-1). Liquid chromatographic separation and post-column detection using the tyrosinase electrode is shown for spiked samples of serum.

Biosensing Techniques↗

Determination of organophosphorus pesticides using membrane-assisted solvent extraction combined with large volume injection-gas chromatography-mass spectrometric detection.

Eight organophosphorus pesticides (parathion-methyl, fenitrothion, malathion, fenthion, bromophos, bromophos-ethyl, fenamiphos and ethion) in aqueous samples were analysed by means of membrane-assisted solvent extraction. First a 20 ml extraction vial was filled with 15 ml of aqueous sample. Then the membrane bag consisting of nonporous polypropylene was put into the vial and filled with 800 microl of organic solvent. The analytes were separated from the aqueous layer by transporting them through the membrane material into the small amount of solvent. The technique was fully automated and successfully combinable with large volume extraction and GC-MS. To achieve an optimum performance several extraction conditions were investigated. Cyclohexane was chosen as acceptor phase. Then the impact of salt, methanol, pH value, as well as working parameters like stirring rate of the agitator and extraction time, were studied. Moreover, the influence of matrix effects was examined by adding different concentrations of humic acid sodium salt. Detection limits in the ng/l level were achieved using large volume injection with the injecting volume of 100 microl. The recovery values ranged from 47 to 100% and the relative standard deviation for three standard measurements was between 4 and 12% (except for bromophos-ethyl: 22%). The linear dynamic range was between 0.001 and 70 microg/l. The applicability of the method to real samples was tested by spiking the eight organophosphorus pesticides to red wine, white wine and apple juice samples.

Beverages↗

Selective detection of chlorine-containing compounds by gas chromatography/chemical reaction interface mass spectrometry.

Chemical reaction interface mass spectrometry (CRIMS) was studied as a gas chromatographic detection technique for chlorine-containing compounds. Both SO2 and HBr were tested as reactant gases. With SO2, a detection limit of 50 pg of diazepam and a linear range of 4 orders of magnitude were achieved, and the experimental data were reproducible. With HBr, the detection limit was 10 ng of diazepam and the linear dynamic range was only 2 orders of magnitude. The possible pharmacological application of CRIMS was studied using urine spiked with diazepam and several of its metabolites, and the results show CRIMS to be a simple but potentially powerful method in drug metabolism studies.

Acids↗

Microsequential injection: anion separations using 'lab-on-valve' coupled with capillary electrophoresis.

Microsequential injection (microSI) has been successfully coupled with capillary electrophoresis (CE). Presented is the microSI-CE system, interfaced with an integrated Lab-on-Valve (LOV) manifold that provides an efficient sample delivery conduit and a versatile means of sample pretreatment along with total automation of the separation process. Programmable microSI protocols control all critical system peripherals to perform various types of CE sample injections automatically such as electrokinetic (EK) injection, hydrodynamic (HD) injection, and head column field amplification (HCFA) sample stacking injection. Novel features of the microSI-CE technique are demonstrated on assays of samples containing 10 anions that had been used previously as a model system. Calibration studies by EK sample injection yielded linear concentration ranges of 0.5-3.0 mM with linear regression responses of r2 = 0.9999 for both chloride and sulfate using conductivity corrected peak area (CCPA) as concentration responses. Calibration using an internal standard was studied at the same concentration range giving r2 = 0.9992 for both chloride and sulfate and r2 = 0.9997 for both when CCPA correction was deployed. With HCFA sample stacking injection, a linear concentration dynamic range of 0.034-3.419 mM for chloride and 0.014-1.408 mM for sulfate were produced with linear regression responses of r2 = 0.9999 for chloride and r2 = 0.9998 for sulfate.

Anions↗

Noise induced complexity: from subthreshold oscillations to spiking in coupled excitable systems.

We study the stochastic dynamics of an ensemble of N globally coupled excitable elements. Each element is modeled by a FitzHugh-Nagumo oscillator and is disturbed by independent Gaussian noise. In simulations of the Langevin dynamics we characterize the collective behavior of the ensemble in terms of its mean field and show that with the increase of noise the mean field displays a transition from a steady equilibrium to global oscillations and then, for sufficiently large noise, back to another equilibrium. In the course of this transition diverse regimes of collective dynamics ranging from periodic subthreshold oscillations to large-amplitude oscillations and chaos are observed. In order to understand the details and mechanisms of these noise-induced dynamics we consider the thermodynamic limit N-->infinity of the ensemble, and derive the cumulant expansion describing temporal evolution of the mean field fluctuations. In Gaussian approximation this allows us to perform the bifurcation analysis; its results are in good qualitative agreement with dynamical scenarios observed in the stochastic simulations of large ensembles.

Action Potentials↗