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Responses of primate spinomesencephalic tract cells to intradermal capsaicin.

The responses of 32 spinomesencephalic tract cells to intradermal capsaicin were examined in anesthetized monkeys. Wide dynamic range (n = 20) and nociceptive specific (n = 6) cells showed two types of excitatory responses to intradermal injection of capsaicin. The first excitatory response shown by the majority of wide dynamic range (n = 13) and nociceptive specific (n = 4) cells was consistent with their sensitization by capsaicin. The cells showed an acute and prolonged increase in ongoing activity with capsaicin injection. Responses to mechanical stimuli were substantially increased after capsaicin and an expansion of receptive field was often observed. The responses of the same cells to excitatory amino acid agonists applied locally by iontophoresis also increased. All cells showing sensitization were antidromically activated from periaqueductal gray regions dorsal to the sulcus limitans. Electrical stimulation at these sites did not affect the ongoing or evoked discharges of these cells. The second excitatory response of wide dynamic range (n = 5) and nociceptive specific (n = 1) cells was a novel pattern not consistent with sensitization. These cells nevertheless showed an acute and prolonged increase in background activity after capsaicin injection. Yet, there was no change or a decrease in responses to cutaneous stimuli, no evidence for change in receptive field size and no increase in responses to locally released excitatory amino acids. These cells projected to regions in the periaqueductal gray ventral to the sulcus limitans. Electrical stimulation at these sites produced a decrease in spontaneous activity of the same cell. Low threshold mesencephalic-projecting neurons (n = 6) showed a single inhibitory pattern (n = 4) of responses to capsaicin. The injection produced an acute decrease in spontaneous activity that was sustained for at least 30 min after injection. The responses to cutaneous stimuli and to excitatory amino acids were also substantially reduced. Low threshold cells were found that projected to both dorsal-lateral and ventral-lateral regions of the periaqueductal gray. In summary, three patterns of responses shown by primate spinomesencephalic tract cells to intradermal capsaicin appear dependent on the functional regions of the periaqueductal gray to which they project. These results suggest that inputs of spinomesencephalic tract neurons to the periaqueductal gray may evoke important components of the systemic response to the neurogenic hyperalgesia produced by intradermal capsaicin.

Animals↗

The effect of monaural middle ear destruction on response properties of neurons in the auditory midbrain of juvenile and adult mice.

This article reviews our studies of the effect of monaural middle ear destruction on midbrain auditory response properties of the laboratory mouse, Mus musculus. Monaural middle ear destruction was performed on juvenile and adult mice and the auditory sensitivity of neurons in the midbrain inferior colliculus (IC) ipsilateral and contralateral to the intact ear was examined 4 weeks later. When stimulated with sound pulses, IC neurons of the control mice typically had lower minimum threshold, larger dynamic range, and sharper frequency tuning curve than IC neurons of the experimental juvenile and adult mice. In the experimental mice, neurons in the ipsilateral IC had significantly longer latency, higher minimum threshold, and smaller dynamic range than neurons in the contralateral IC. When determined at two sound directions (ipsilateral 40 degrees and contralateral 40 degrees to the recording site), IC neurons of the control mice had higher minimum threshold, sharper frequency tuning curve but smaller dynamic range at I-40 degrees than at C-40 degrees . However, these direction-dependent response properties were not observed for IC neurons of the experimental juvenile and adult mice. Clear tonotopic organization was only observed in the IC of the control mice and experimental adult mice but not in the IC of experimental juvenile mice. These different response properties are discussed in relation to the effect of monaural middle ear destruction.

Acoustic Stimulation↗

Naturalistic auditory contrast improves spectrotemporal coding in the cat inferior colliculus.

Statistical analysis of natural sounds and speech reveals logarithmically distributed spectrotemporal modulations that can cover several orders of magnitude. By contrast, most artificial stimuli used to probe auditory function, including pure tones and white noise, have linearly distributed amplitude fluctuations with a limited average dynamic range. Here we explore whether the operating range of the auditory system is physically matched to the statistical structure of natural sounds. We recorded single-unit and multi-unit neuronal activity from the central nucleus of the cat inferior colliculus (ICC) in response to dynamic spectrotemporal sound sequences to determine whether ICC neurons respond preferentially to linear or logarithmic spectrotemporal amplitudes. We varied the intensity, dynamic range, and contrast statistics of these sounds to mimic those of natural and artificial stimuli. ICC neurons exhibited monotonic and nonmonotonic contrast dependencies with increasing dynamic range that were independent of the stimulus intensity. Midbrain neurons had higher firing rates and higher receptive field energies and showed a net improvement in spectrotemporal encoding ability for logarithmic stimuli, with an increase in the mutual information rate of approximately 50% over linear amplitude sounds. This efficient use of logarithmic spectrotemporal modulations by auditory midbrain neurons reflects a neural adaptation to structural regularities in natural sounds and likely underlies human perceptual abilities.

Action Potentials↗

Utilizing human blood plasma for proteomic biomarker discovery.

Candidate proteomic biomarker discovery from human plasma holds both incredible clinical potential as well as significant challenges. The dynamic range of proteins within plasma is known to exceed 10(10), and many potential biomarkers are likely present at lower protein abundances. At present, proteomic based MS analyses provide a dynamic range typically not exceeding approximately 10(3) in a single spectrum, and approximately 10(4)-10(6) when combined with on-line separations (e.g., reversed-phase gradient liquid chromatography), and thus are generally insufficient for low level biomarker detection directly from human plasma. This limitation is providing an impetus for the development of experimental methodologies and strategies to increase the possible number of detections within this biofluid. Discussed is the diversity of available approaches currently used by our laboratory and others to utilize human plasma as a viable medium for biomarker discovery. Various separation, depletion, enrichment, and quantitative efforts as well as recent improvements in MS capabilities have resulted in measurable improvements in the detection and identification of lower abundance proteins (by approximately 10-10(2)). Despite these improvements, further advances are needed to provide a basis for discovery of candidate biomarkers at very low levels. Continued development of depletion and enrichment techniques, coupled with improved pre-MS separations (both at the protein and peptide level) holds promise in extending the dynamic range of proteomic analysis.

Biomarkers↗

Development of a PCR-compatible enrichment medium for Yersinia enterocolitica: amplification precision and dynamic detection range during cultivation.

A Yersinia PCR-Compatible Enrichment (YPCE) medium was developed, which removes the necessity for sample pretreatment before PCR-based detection of Yersinia enterocolitica. The medium was designed through a sequence of independent screening and factorial design experiments to study the PCR inhibition and growth characteristics of medium components. The compatibility of the YPCE medium was evaluated using real-time PCR. The real-time PCR assay, based on the fluorescent double-stranded DNA binding dye SYBR green, generated approximately a 4-log linear range of amplification and in the range of 10(5)-10(8) (CFU/ml), the coefficient of variation <5%. When a background flora was present at concentrations > or = 10(6) (CFU/ml), the DNA amplification was influenced and a change in the log-linear slope leading to a lower amplification efficiency was observed. To study the dynamic detection range and relative amplification precision during enrichment. Y. enterocolitica and background flora were inoculated at various concentrations. It was possible to detect inoculation concentrations of 10(1) (CFU/ml) Y. enterocolitica in the presence of at least an inoculation concentration of 10(3) (CFU/ml) of an undefined background flora and the optimal conditions for sample withdrawal was in the range of 9 to 18 h enrichment. The YPCE medium can, especially for swab samples, form part of a simple analysis procedure allowing high throughput PCR.

Buffers↗

Relations between audiometrical thresholds of potential cochlear implant patients and their performance in preoperative psycho-physical tests with electrical stimulation.

Part of our selection procedure for cochlear implant candidates is the preoperative electrical stimulation of the cochlea. With a transtympanic needle, placed as closely as possible to the round window, the dynamic range for a number of frequencies is measured and a frequency identification test and a temporal difference limen test are carried out. Our first series of preoperative tests suggests a relationship between the audiometrical pure-tone thresholds, the dynamic range of electrical stimulation and the performance on the temporal difference and frequency identification test. For the preoperative electrical stimulation tests, a relatively good pure-tone average may indicate a wide dynamic range, a short temporal difference limen and a high score in the frequency identification test.

Audiometry, Evoked Response↗

Intracochlear factors contributing to psychophysical percepts following cochlear implantation.

The performance of cochlear implant patients may be related to intracochlear, histopathological factors. We have performed detailed post-mortem examinations of five human, implanted cochleas and for each electrode correlated the psychophysical threshold, comfortable level and dynamic range with spiral ganglion cell survival, presence of fibrous tissue and/or new bone, and distance between the centers of the electrode bands and Rosenthal's canal. The psychophysical parameters were strongly interrelated. Threshold and comfort levels correlated with the distance between the electrodes and Rosenthal's canal. Threshold levels also correlated with the presence of intracochlear fibrous tissue and new bone, especially with the former. The dynamic range showed a negative correlation with intracochlear pathology, especially with new bone. Comfort levels and dynamic range were related to spiral ganglion cell survival. The distance between the electrodes and the modiolus increased with increasing levels of fibrous tissue and new bone. Spiral ganglion cell survival was decreased with increasing levels of fibrous tissue and new bone.

Adult↗

Plasma SIV RNA viral load determination by real-time quantification of product generation in reverse transcriptase-polymerase chain reaction.

Internally controlled RT-PCR methods (QC-RT-PCR) for quantification of SIV RNA are effective, but are relatively cumbersome, expensive, and time and labor intensive. For greater throughput and efficiency, we have developed a method for quantification of plasma SIV RNA levels by real-time RT-PCR using the Applied Biosystems Prism 7700 sequence detection system. This assay format allows real-time kinetic analysis of PCR product generation, providing a broad linear dynamic range and ensuring that quantification is based on analysis during the exponential phase of amplification, regardless of the input template copy number. Simultaneous amplification and analysis eliminates any requirement for handling amplified products, increasing throughput and eliminating a potential source of assay contamination. The assay we have developed for quantification of SIV RNA has a nominal threshold sensitivity of 300 copy Eq/ml of plasma, although as little as 10 copy Eq/reaction of SIV RNA template can be detected. The linear dynamic range is in excess of 5 logs. Interassay reproducibility averages 25% (coefficient of variation), based on studies of extraction and analysis of replicate aliquots of the same plasma specimens. The combination of sensitivity, precision, and broad dynamic range allows reliable quantification of viral load even during dynamic phases of SIV infection, such as through the onset and resolution of primary infection, or during treatment with antiretroviral agents. The primer-probe combinations we have developed allow quantification of SIV isolates most commonly used for experimental studies. Availability of this assay should greatly facilitate studies of basic pathogenesis and evaluation of therapeutic and prophylactic approaches in the SIV-infected macaque.

Animals↗

Apparent auditory deprivation effects of late onset: the role of presentation level.

Silman and colleagues [J. Acoust. Soc. Am. 76, 1347-1362 (1984)] have reported an apparent effect of late auditory deprivation; this presents as loss of discrimination over time in the unaided ear of individuals using a single hearing aid fitted in middle age. In a replication of the basic effect, the influence of presentation level was examined in 24 monaurally aided subjects. The effect was reversed at presentation levels below about 75 dB SPL. The ear that is normally aided performs better at high presentation levels, while, at lower presentation levels, the converse is true. Thus it appears that a form of selective adjustment takes place in a particular part of the dynamic range, at least in ears with a dynamic range limited by a sensory hearing loss. If this interpretation is correct, there are important implications for research on perceptual learning and for the time course of evaluation in hearing aid provision.

Audiometry, Pure-Tone↗

Power Doppler US: a potentially useful alternative to mean frequency-based color Doppler US.

PURPOSE: The authors present a preliminary report to demonstrate a new color Doppler (CD) ultrasonography (US) technique called power Doppler (PD), which displays the total integrated Doppler power in color, and to compare PD with CD imaging, which generally displays an estimate of the mean Doppler frequency shift. MATERIALS AND METHODS: Two standard commercial US scanners that encode the integrated power in the Doppler signal in color were used to demonstrate PD. A standard nonflow-containing US phantom, a normal right kidney, and a torsive and normal contralateral testis were scanned in the power mode. In the phantom and kidney, results with CD and PD were directly compared. RESULTS: PD does not alias, is relatively angle independent, and displays background noise in a way that increases the usable dynamic range of a US scanner. This extended dynamic range should increase machine sensitivity and may demonstrate increased flow in certain circumstances. CONCLUSION: PD is a new CD imaging imaging mode that might be superior to CD in some cases.

Humans↗

FAST sequences optimization for contrast media pharmacokinetic quantification in tissue.

The purpose of this study was to investigate the influence of the fast gradient-recalled echo (GRE) sequence parameters on the contrast dynamic range and signal sensitivity, to optimize the magnetic resonance (MR) sequence for contrast media pharmacokinetic assessment. Effects of the fast low-angle shot (FLASH), Fast acquisition at steady rate (FAST), and radiofrequency-spoiled (RF)-FAST sequence parameters were studied in vitro. The FAST sequence had the highest sensitivity in low gadolinium (Gd) concentration. The FLASH and RF-FAST sequences had a larger contrast dynamic range, but the FLASH images contained side band artifacts. Increasing the flip angle to 90 degrees raised the sensitivity of the FAST sequence and the contrast dynamic range of the RF-FAST sequence. The shortest possible TE was optimal for both contrast dynamics and imaging time. TI had an influence on the sensitivity of the FAST sequence only for small acquisition matrices. This study indicates the optimal parameters for contrast dynamics (RF-FAST, 90 degrees flip angle, shortest possible TE) and sensitivity (FAST, 90 degrees flip angle, long TI(eff)).

Contrast Media↗

Intensity discrimination with cochlear implants.

Intensity difference limens were measured for various frequencies and intensities of sinusoidal and pulsatile electrical stimulation in monkeys with electrodes implanted in the scala tympani, scala vestibuli, modiolus, or middle ear. Difference limens decreased, as a function of initial stimulus intensity, from values of 1.5-3 dB near threshold to as low as 0.5 dB near the upper limit of the dynamic range. If sensation level was held constant, difference limens decreased as a function of frequency up to about 500 Hz, and then remained constant. They were similar across a variety of electrode placements and separations if differences in threshold and dynamic range were taken into account. However, difference limens measured in severely damaged ears were slightly smaller than those in moderately damaged ears. The near miss to Weber's law, characteristic of acoustic difference limens, was not seen with electrical stimuli. Differences limens for electrical stimuli were roughly one-half those for acoustic stimuli; thus, part of the deficit in dynamic range for electrical stimulation compared with acoustic stimulation is countered by the smaller intensity differences limens for electrical stimuli.

Acoustic Stimulation↗

Life, evolution, and the pursuit of single photon sensitivity.

Evolutionary forces have designed a large family of rod and cone photoreceptors, each member of which suits the lifestyle requirements and circadian patterns of a particular species. The three-segment architecture of signal transduction is conspicuous in the biochemistry of photoreceptors and supports their demonstrated properties of extreme sensitivity, low noise levels, extended dynamic range, and light adaptation. The designs elaborated by evolution reflect a gradual process of modification, with the sequential elaboration of layers of control and refinements in control. The end results of this long evolutionary labor are the functional efficiency and dynamic range that give the rod its utility. Our conceptual problems in deriving observed rod properties from the collective features of known rod gene products may well give way when we have learned more about the true composition and topology of the outer segment gene set and both bound and free nucleotide concentrations. The invertebrates have developed alternative solutions to the problems of photoreceptor sensitivity and wide dynamic range. The vertebrate rod represents a truly optimized way to capture and interpret low-intensity photon signals. One may anticipate, with some enthusiasm, those molecular and kinetic data that will permit an understanding of how cones differ from rods and how release from the requirement for single photon detection has shaped the design of this wavelength-specific companion photoreceptor. The utilization by evolution of the three-segment architecture of GTP-dependent signal transduction for other modalities of sensory perception, such as olfaction (Lancet et al., this volume) and gustation (Jones et al., this volume), is certainly a reasonable and successful choice.(ABSTRACT TRUNCATED AT 250 WORDS)

3',5'-Cyclic-GMP Phosphodiesterases↗

Suppression of noxiously evoked WDR dorsal horn neuronal activity by spinally administered morphine.

The present study was carried out in order to examine the ability of spinally administered morphine to suppress noxiously evoked activity of wide-dynamic-range (WDR) neurons in the dorsal horn of the spinal cord in decerebrate, spinal cord-transected cats. All cells (n = 25) responded maximally to high-intensity noxious heat stimulation (51 degrees C) and were classified as wide dynamic range neurons. The spinal administration of 0.1 mg of morphine caused a significant reduction of noxiously evoked activity but did not significantly change spontaneous activity. The 0.25-mg dose caused a significant reduction of both types of activity. Thirty minutes after spinal administration, 0.1 mg of morphine caused a 27% reduction of spontaneous activity and a 43% reduction of noxiously evoked activity. The 0.25-mg dose reduced spontaneous activity by 44% and the evoked activity by 70%. Naloxone partially reversed the morphine-induced neuronal suppression. In addition, in the four neurons in which it was tried, spinally administered epinephrine was found to further suppress the remaining neuronal activity following the spinal morphine effect. These results demonstrate for the first time that spinally administered morphine is capable of suppressing noxiously evoked activity of wide-dynamic-range neurons in the dorsal horn of the spinal cord. They also demonstrate the dose-dependent nature of this effect and the potential importance of the interaction between morphine and adrenergic agonists in blocking information about noxious events. This information provides a probable mechanism of action for spinal opiate analgesia.

Action Potentials↗

Mechanical response properties of ventroposterior medial thalamic neurons in the alert monkey.

The properties of trigeminal cutaneous thalamic neurons were explored in alert cynomolgous monkeys to determine receptive field and response characteristics. Two monkeys received juice reward for sitting quietly while an investigator probed the monkey's face with mechanical stimuli. Extracellular single unit recordings were made from the ventroposterior medial thalamic nucleus (VPM), and mechanical response properties were evaluated for each cell having an extraoral cutaneous receptive field. Of 89 cells examined, 90% responded best to innocuous tactile stimulation, and were classified as low threshold. The other 10%, classified as wide dynamic range, showed a graded response to increasingly intense stimuli, with a maximum discharge to noxious pinch. Of the low threshold neurons, most exhibited excitatory responses, with about half being rapidly adapting and the others slowly adapting. The spontaneous activity of 11% of the low threshold neurons was inhibited by stimulation of the neuron's receptive field. There was no systematic difference in receptive field size for the various types of neurons, but the receptive fields of wide dynamic range cells were smaller than those previously observed in trigeminopthalamic neurons of the medullary dorsal horn. The wide dynamic range and inhibitory low threshold neurons were located primarily in the caudal third of VPM, while the excitatory low threshold neurons were located throughout. In summary, response characteristics of VPM neurons show more diversity in the alert monkey than has been reported in paralyzed and/or anesthetized animals.

Adaptation, Physiological↗

Electrospray ionization-Fourier transform ion cyclotron mass spectrometry using ion preselection and external accumulation for ultrahigh sensitivity.

The dynamic range of Fourier transform ion cyclotron mass spectrometry (FTICR) is typically limited by the useful charge capacity of an FTICR cell (to approximately 10(6) to 10(7) elementary charges) and the minimum number of ions required to produce a useful signal (approximately 10(2) elementary charges). We show that the expansion of the dynamic range by 2 orders of magnitude can be achieved by preselecting lower abundance species in a quadrupole interface to an electrospray ionization (ESI) source. Ion preselection is then followed by ion accumulation in external to the FTICR cell a linear (2-D) quadrupole trap and subsequent transfer to the region of high magnetic field for gated trapping in the FTICR cell. Two modes of ion preselection, using either the quadrupole filtering mode or rf-only dipolar excitation, were studied and mass resolutions of 30 to 100 were achieved for selective external ion accumulation of peptides and proteins with molecular weights ranging from 500 to 17,000 Da. The ability to selectively eject the most abundant species before trapping in the FTICR has enormous practical benefits for increasing the sensitivity and dynamic range of measurements.

Algorithms↗

Hepatitis C virus RNA assays: a comparison of SuperQuant and Monitor.

Hepatitis C RNA testing has been used extensively to assess the efficacy of antiviral therapy and has increasingly become an integral part of clinical management of patients with chronic hepatitis C. A variety of commercially available hepatitis C virus (HCV) RNA tests are used to detect HCV RNA qualitatively or quantitatively. These commercial tests have fundamental differences that are reflected on the values they generate. We compared two widely used assays, HCV SuperQuant (SQ) and Amplicor HCV Monitor (M1 and M2), in sera of patients with chronic hepatitis C. A total of 506 sera from 79 patients were tested with both assays. The data were logarithmically transformed and analyzed by linear regression and measurement of agreement. Two hundred thirty-eight sera had HCV RNA values within the dynamic range of both assays. The correlation between the assays was fair, with a correlation coefficient (r) of 0.699. Overall, SQ generated higher values than M1 with a mean difference of 0.558 log (SD = 0.624). One hundred ninety-four (38%) and 121 (24%) of the sera were below the dynamic range of M1 and SQ, respectively. Seventy-three sera, undetectable by M1, were positive by SQ. The Amplicor HCV Monitor 2.0 (M2) was performed in 66 sera. All were positive by SQ and M2, but only 38 were within the dynamic range of M1. The correlations between these tests were good (r = 0.68-0.78), but the agreement was rather poor. In conclusion, this study confirms that both SQ and M2 are more sensitive than M1. Additionally, our results show rather poor agreements between these assays. The recent attempts in standardizing the reporting of these assays should make their results more easily interchangeable.

Adult↗

Status of complete proteome analysis by mass spectrometry: SILAC labeled yeast as a model system.

BACKGROUND: Mass spectrometry has become a powerful tool for the analysis of large numbers of proteins in complex samples, enabling much of proteomics. Due to various analytical challenges, so far no proteome has been sequenced completely. O'Shea, Weissman and co-workers have recently determined the copy number of yeast proteins, making this proteome an excellent model system to study factors affecting coverage. RESULTS: To probe the yeast proteome in depth and determine factors currently preventing complete analysis, we grew yeast cells, extracted proteins and separated them by one-dimensional gel electrophoresis. Peptides resulting from trypsin digestion were analyzed by liquid chromatography mass spectrometry on a linear ion trap-Fourier transform mass spectrometer with very high mass accuracy and sequencing speed. We achieved unambiguous identification of more than 2,000 proteins, including very low abundant ones. Effective dynamic range was limited to about 1,000 and effective sensitivity to about 500 femtomoles, far from the subfemtomole sensitivity possible with single proteins. We used SILAC (stable isotope labeling by amino acids in cell culture) to generate one-to-one pairs of true peptide signals and investigated if sensitivity, sequencing speed or dynamic range were limiting the analysis. CONCLUSION: Advanced mass spectrometry methods can unambiguously identify more than 2,000 proteins in a single proteome. Complex mixture analysis is not limited by sensitivity but by a combination of dynamic range (high abundance peptides preventing sequencing of low abundance ones) and by effective sequencing speed. Substantially increased coverage of the yeast proteome appears feasible with further development in software and instrumentation.

Amino Acid Sequence↗