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At least 1,225 records · Page 68Linked to original sources
PubMed1999

A neural network for enhancing boundaries and surfaces in synthetic aperture radar images.

A neural network system for boundary segmentation and surface representation, inspired by a new local-circuit model of visual processing in the cerebral cortex, is used to enhance images of range data gathered by a synthetic aperture radar (SAR) sensor. Boundary segmentation is accomplished by an improved Boundary Contour System (BCS) model which completes coherent boundaries that retain their sensitivity to image contrasts and locations. A Feature Contour System (FCS) model compensates for local contrast variations and uses the compensated signals to diffusively fill-in surface regions within the BCS boundaries. Image noise pixels that are not supported by BCS boundaries are hereby eliminated. More generally, BCS/FCS processing normalizes input dynamic range, reduces noise, and enhances contrasts between surface regions. BCS/FCS processing hereby makes structures such as motor vehicles, roads, and buildings more salient to human observers than in original imagery. The new BCS model improves image enhancement with significant reductions in processing time and complexity over previous BCS applications. The new system also outperforms several established techniques for image enhancement.

Journal Article↗
PubMed2000

Chemical and physiological characterization of fluo-4 Ca(2+)-indicator dyes.

We have developed fluo-4, a new fluorescent dye for quantifying cellular Ca2+ concentrations in the 100 nM to 1 microM range. Fluo-4 is similar in structure and spectral properties to the widely used fluorescent Ca(2+)-indicator dye, fluo-3, but it has certain advantages over fluo-3. Due to its greater absorption near 488 nm, fluo-4 offers substantially brighter fluorescence emission when used with excitation by argon-ion laser or other sources in conjunction with the standard fluorescein filter set. In vitro, fluo-4 exhibited high fluorescence emission, a high rate of cell permeation, and a large dynamic range for reporting [Ca2+] around a Kd(Ca2+) of 345 nM. We have also developed several Ca(2+)-indicators related to fluo-4 having lower affinities for Ca2+ that are useful in cellular studies requiring quantification of higher [Ca2+]. In a variety of physiological studies of live cells, fluo-4 labeled cells more brightly than did fluo-3, when challenged with procedures designed to elevate calcium levels. Fluo-4 is well suited for photometric and imaging applications that make use of confocal laser scanning microscopy, flow cytometry, or spectrofluorometry, or in fluorometric high-throughput microplate screening assays. Because of its higher fluorescence emission intensity, fluo-4 can be used at lower intracellular concentrations, making its use a less invasive practice.

3T3 Cells↗
PubMed1992

[Evoked oto-acoustic emissions in retrocochlear deafness].

In order to give a contribution to the genesis of the EOAEs, we have recorded the echoes in 6 subjects affected with unilateral acoustic neuroma, surgically proven (I. group); 11 subjects with sensorineural hearing loss with electrophysiological or balance signs of retrocochlear impairment (II. group) and 10 normally hearing young subjects as control group. In this study EOAEs have been recorded in response respectively to 3 ms 1000 Hz tone-burst and to 200-5000 filtered click, both sending with 2048 stimuli, at a repetition rate of 21 pps and different stimulus intensities ranging from 40 db SPL to saturation threshold in 10 db steps. The results have shown that the EOAEs were present in half of the tested patients in response to the clicks in both I. and II. group and in all the ears to the tone-burst although the 1.0 KHz audiometric threshold mean were > 65 db HL in I. group and in a third of the subjects in the II. group. The properties of EOAEs (detection and saturation threshold, dynamic range and duration) depended on the degree of hearing loss. These results suggest that acoustic tumor could determine a wallerian degeneration of the external fibers of the VIII nerve and consequently a dysfunction of the correspondent external hair cells of the cochlea, while in the II. group a contemporary or secondary involvement of the cochlear structures is possible.

Adult↗
PubMed1990

Distortion product emissions in humans. I. Basic properties in normally hearing subjects.

Distortion product otoacoustic emissions (DPEs) at the 2f1-f2 frequency were recorded from 44 normal ears in response to equilevel primary tones. Detailed testing included the recording of DPE "audiograms" in 100-Hz steps from 1 to 8 kHz at three primary-tone levels (65, 75, and 85 dB sound pressure level [SPL]). In addition, response-growth or input-output (I/O) functions depicting the relationship of the amplitudes of DPEs to primary-tone levels, ranging from 25 to 85 dB SPL in 5-dB steps, were also tested for 11 frequencies distributed at quarter-octave intervals over the identical frequency range. The average DPE "audiogram" illustrating the frequency response of these emissions demonstrated a bilobed contour having a low-frequency maximum at approximately 1.5 kHz and a high-frequency peak that plateaued at about 5.5 kHz. The two maximum regions were separated by a minimum around 2.5 kHz. Depending on the frequency region, the average I/O functions exhibited detection "thresholds" at 3 dB above the noise floor at primary levels between 35 and 45 dB sound pressure level. The dynamic range of the emitted response between detection "threshold" and maximum amplitude varied over a 40-dB extent of the stimulus-level dimension. Approximately one third of the ears exhibited irregular DPE "audiograms" in which emitted responses were significantly reduced in restricted regions tested by low, medium, or high frequencies. When the 44 ears were separated into two groups representing more-normal and less-normal responses, the irregular "normal" ears demonstrated increased variability, especially in high-frequency regions. Mean age did not explain the differences noted between the two types of normally hearing subjects. However, across ears, DPE amplitudes and "thresholds" for the highest frequencies tested were correlated significantly with age in that the oldest individuals showed higher "thresholds" and lower amplitudes.

Acoustic Impedance Tests↗
PubMed2006

A high-throughput, fully automated liquid/liquid extraction liquid chromatography/mass spectrometry method for the quantitation of a new investigational drug ABT-869 and its metabolite A-849529 in human plasma samples.

ABT-869 is a novel ATP-competitive inhibitor for all the vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF) receptor tyrosine kinases (RTKs). It is one of the oncology drugs in development at Abbott Laboratories and has great potential for enhanced anti-tumor efficacy as well as activity in a broad range of human cancers. We report here an accurate, precise and rugged liquid chromatography/mass spectrometry (LC/MS/MS) assay for the quantitative measurement of ABT-869 and its acid metabolite A-849529. A fully automated 96-well liquid/liquid extraction method was achieved utilizing a Hamilton liquid handler. The only manual intervention required prior to LC/MS/MS injection is to transfer the 96-well plate to a drying rack to dry the extracts then transfer the plate back to the Hamilton for robotic reconstitution. The linear dynamic ranges were from 1.1 to 598.8 ng/mL for ABT-869 and from 1.1 to 605.8 ng/mL for A-849529. The coefficient of determination (r2) for all analytes was greater than 0.9995. For the drug ABT-869, the intra-assay coefficient of variance (CV) was between 0.4% and 3.7% and the inter-assay CV was between 0.9% and 2.8%. The inter-assay mean accuracy, expressed as percent of theoretical, was between 96.8% and 102.2%. For the metabolite A-849529, the intra-assay CV was between 0.5% and 5.1% and the inter-assay CV was between 0.8% and 4.9%. The inter-assay mean accuracy, expressed as percent of theoretical, was between 96.9% and 100.6%.

Blood Chemical Analysis↗
PubMed1993

Directional characteristics of tuberous electroreceptors in the weakly electric fish, Hypopomus (Gymnotiformes).

This paper is an electrophysiological study of the directionality of the tuberous electroreceptors of weakly electric fish. We recorded from two classes of tuberous electroreceptors known for pulse gymnotiforms: Burst Duration Coders (BDCs), and Pulse Markers (PMs). Both code for stimulus amplitude, although the dynamic range for BDCs is greater, and both exhibit strong directional preferences. Polar plots of spike number (for BDCs) or spike threshold (for PMs) versus electric field azimuth, are figure-8 shaped with two asymmetrical, elliptical lobes separated by 180 degrees. The best azimuth of these two types of receptors from a given body region correlate with each other and with measures of best azimuth for transepidermal current flow. The shape and asymmetry of the directionality profiles appear to be caused by filter dynamics of the receptors. Pulse Markers are located on the anterior part of the body surface while Burst Duration Coders are located all over. The best directions of receptors in the anterior third of the body vary systematically with location from 0 degrees to 180 degrees. This region is probably critical for determining the direction of local electric fields. Together these receptors provide the CNS with sufficient information to construct a map of horizontal plane electric field directions.

Adaptation, Physiological↗
PubMed2003

Trace level determination of lead in solid samples by UV laser ablation and laser-enhanced ionization detection.

Laser-enhanced ionization was investigated as a detection technique for trace elemental analysis of solid samples by laser ablation. Laser ablation of aluminum samples was performed in an ablation cell, and the ablated material was carried by a flow of gas to a miniature LEI flame where Pb was detected. This decoupling of ablation cell and detector allowed the independent optimization of vaporization and detection processes. We have investigated the different excitation schemes for Pb and uncovered five new LEI-active transitions in the visible range. We have demonstrated that the use of an argon-oxygen/acetylene flame sheathed with argon resulted in the elimination of background interference from the two-photon ionization of nitric oxide. We have shown that the use of helium as a carrier gas results in a higher ablation yield and lower pulse-to-pulse variations in LEI signal and in better analytical figures of merit. We have characterized the performance of the technique in terms of detection limits and dynamic range, and we have obtained a detection limit of 60 ng/g for the determination of Pb in high purity aluminum.

Journal Article↗
PubMed1992

A computer model of a cochlear-nucleus stellate cell: responses to amplitude-modulated and pure-tone stimuli.

A computer model of a ventral-cochlear-nucleus (VCN) stellate cell with chop-S type response properties is presented and evaluated. The model is based on a simplified model of spike generation preceded by a stage that simulates dendritic low-pass filtering. Input to the model is in the form of simulated auditory-nerve spikes produced by a model of the auditory periphery [Meddis and Hewitt, J. Acoust. Soc. Am. 89, 2866-2882 (1991)]. Outputs from the stellate-cell model are shown to qualitatively replicate a wide range of typical in vivo responses. These include: (a) realistic onset and steady-state rate-level functions, (b) "chopper"-type post-stimulus time histogram responses; (c) typical "chop-S"-type neuron responses characterized by a low coefficient of variance (CV less than 0.3) of interspike intervals as a function of time; (d) level-dependent amplitude-modulation transfer functions; (e) intrinsic oscillations in responses to pure-tone stimuli; (f) amplitude-modulation encoding over a wide dynamic range; and (g) frequency-limited phase locking to pure tones. It is shown that these responses can be explained primarily by the membrane properties of the cells. More specifically, how the model encodes signal amplitude modulation was studied and an explanation was suggested for the generation of the bandpass modulation transfer functions. Such functions are observed neurally in response to amplitude-modulated stimuli presented at moderate to high signal levels.

Animals↗
PubMed2002

Quantitative reverse transcription-PCR measurement of tissue factor mRNA in glioma.

BACKGROUND: Initiation of the coagulation serine protease cascade in mammalian cells is mediated by tissue factor (TF), which is a cell surface receptor and cofactor for coagulation factor VII (FVII) and its activated form FVII (FVIIa). Increasing evidence suggests that TF is expressed in a wide range of cancer cells and plays important roles in cancer progression and metastasis. In this study, we investigated the association between the expression level of TF transcript and histologic features of glioma. METHODS: RNA was extracted from normal brain tissues and glioma tissues. We developed and validated a real-time quantitative reverse transcription (RT)-PCR assay, based on fluorescent TaqMan methodology, to quantify TF gene expression and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) at the mRNA level in human glioma. RESULTS: The dynamic range of the assay was 10(3)-10(8) copy/microg RNA. The relationship between Ct and log starting concentration was linear (r2 > or = 0.99). The mean expression of TF in healthy brain tissue was 6.2 x 10(3) copy/microg RNA. Overexpression of TF was found in 42 brain glioma samples, mean value is 2.9 x 10(6) copy/microg RNA. CONCLUSIONS: TF mRNA transcript is expressed in glioma and the level of expression correlates with histologic grade of malignancy. This new simple, rapid, semiautomated assay is a major alternative to Northern blot and competitive quantitative PCR for gene alteration analysis in human tumors and may be a powerful tool for large randomized, prospective cooperative group trials and support future TF-based clinical applications.

Biomarkers, Tumor↗
PubMed2001

Use of dynamically coated capillaries for the routine analysis of methamphetamine, amphetamine, MDA, MDMA, MDEA, and cocaine using capillary electrophoresis.

A rapid, accurate, precise, reproducible, economical, and environmentally gentle method using capillary electrophoresis (CE) is presented for the routine analysis of methamphetamine, amphetamine, MDA, MDMA, MDEA, and cocaine in seized drugs. The methodology uses a 32 cm by 50 microm capillary (length to detector 23.5 cm) with a commercially available buffer kit and diode array UV detection. Dynamic coating of the capillary surface is accomplished by flushing with base for 1 min, a proprietary polycation for 1 min, and then a proprietary polyanion for 2 min. This approach provides a relatively high and stable electroosmotic flow (EOF), even at low pHs. The background electrolyte (BGE) contains 75 mM phosphate buffer (pH 2.5) with the same polyanion as above. Using this methodology, amphetamine, methamphetamine, MDA, MDMA, MDEA, and an internal standard (n-butylamphetamine) are baseline resolved in less than 5 min. The run-to-run migration time %RSDs and peak area %RSDs are typically <0.3% and <2.1%, respectively. The day-to-day and capillary-to-capillary migration time %RSDs are <1.5% and <2.1%, respectively. The %RSDs of the relative migration times compared with the internal standard on a day-to-day and capillary-to-capillary basis are <0.2% and <0.06%, respectively. The linear dynamic range using peak areas range from 0.003 to 0.10 mg/mL. The correlation coefficients are >0.9998, with all calibration curves passing at or near the origin. Similar data are obtained for cocaine and its internal standard henyltoloxamine. None of the compounds usually encountered in illicit samples interfere with the target compound (e.g., methamphetamine and cocaine) or the internal standard. Quantitative results for synthetic mixtures and seized exhibits are in good agreement with actual values, and also with results obtained from other techniques. The relatively high EOF for the dynamically coated capillary system allows for the screening of basic, acidic, and neutral adulterants in drug seizures; identification is facilitated by the use of automated UV library searches.

Journal Article↗
PubMed1989

Auditory filter characteristics and consonant recognition for hearing-impaired listeners.

To examine the association between frequency resolution and speech recognition, auditory filter parameters and stop-consonant recognition were determined for 9 normal-hearing and 24 hearing-impaired subjects. In an earlier investigation, the relationship between stop-consonant recognition and the articulation index (AI) had been established on normal-hearing listeners. Based on AI predictions, speech-presentation levels for each subject in this experiment were selected to obtain a wide range of recognition scores. This strategy provides a method of interpreting speech-recognition performance among listeners who vary in magnitude and configuration of hearing loss by assuming that conditions which yield equal audible spectra will result in equivalent performance. It was reasoned that an association between frequency resolution and consonant recognition may be more appropriately estimated if hearing-impaired listeners' performance was measured under conditions that assured equivalent audibility of the speech stimuli. Derived auditory filter parameters indicated that filter widths and dynamic ranges were strongly associated with threshold. Stop-consonant recognition scores for most hearing-impaired listeners were not significantly poorer than predicted by the AI model. Furthermore, differences between observed recognition scores and those predicted by the AI were not associated with auditory filter characteristics, suggesting that frequency resolution and speech recognition may appear to be associated primarily because both are degraded by threshold elevation.

Adolescent↗
PubMed2005

Real-time reverse transcription-PCR quantitation of substance P receptor (NK-1R) mRNA.

The substance P (SP)-preferring receptor, neurokinin-1 receptor (NK-1R), has an important role in inflammation, immune regulation, and viral infection. We applied a newly developed real-time reverse transcription (RT)-PCR assay to quantify NK-1R mRNA in human neuronal cell line (NT-2N), a human B-cell line (IM9), monocyte-derived macrophages (MDM), peripheral blood lymphocytes (PBL), and human astroglioma cells (U87 MG). The NK-1R real-time RT-PCR assay has a sensitivity of 100 mRNA copies, with a dynamic range of detection between 10(2) and 10(7) copies of NK-1R gene transcripts per reaction. This assay is highly reproducible, with an intraassay coefficient variation of threshold cycle (Ct) of less than 1.9%. The NK-1R real-time RT-PCR is highly sensitive for quantitative determination of NK-1R mRNA in human immune cells (MDM and PBL) that express low levels of NK-1R mRNA. In addition, the assay has the ability to accurately quantitate the dynamic changes in NK-1R mRNA expression in interleukin-1beta-stimulated U87 MG. These data indicate that the NK-1R real-time RT-PCR has potential for a wide application in investigation of NK-1R expression at the mRNA level under physiological and pathological conditions in both the central nervous system and the immune system.

Cells, Cultured↗
PubMed2004

Multicenter evaluation of the VERSANT hepatitis B virus DNA 3.0 assay.

The VERSANT hepatitis B virus (HBV) 3.0 Assay (branched DNA [bDNA]) (referred to herein as VERSANT 3.0) was evaluated at four external sites for analytical sensitivity, specificity, reproducibility, linearity of quantification, and limits of detection. In addition, each of the test evaluation sites provided HBV DNA-positive clinical samples that were previously analyzed by one of three commercially available HBV DNA quantitative tests: Digene Hybrid Capture II HBV DNA Test (Digene); VERSANT HBV DNA 1.0 Assay (bDNA) (VERSANT 1.0); and COBAS AMPLICOR HBV Monitor Test (COBAS AMPLICOR). These samples were reexamined using VERSANT 3.0. The results from these studies showed that VERSANT 3.0 has high specificity (99.3%), excellent reproducibility (between-run coefficient of variation [CV] = 1.6 to 9.4%; within-run CV = 6.5 to 20.7%), and a broad linear range of quantification (2.0 x 10(3) to 1.0 x 10(8) HBV DNA copies/ml) that facilitate the monitoring of HBV DNA levels at diagnosis and throughout the course of treatment. In general, correlation was very good between results obtained from clinical samples analyzed by VERSANT 3.0 and the comparative HBV DNA quantitative assays (VERSANT 1.0, R(2) = 0.900; Digene, R(2) = 0.985; COBAS AMPLICOR, R(2) = 0.771). The greatest differences in comparative quantitation occurred at HBV DNA levels approaching the limits of the dynamic ranges for the comparative assays. The performance characteristics of the new VERSANT 3.0 assay demonstrated that it provides a reliable and robust method for routinely monitoring serum HBV DNA levels in assessing disease activity and determining response to antiviral treatment.

DNA, Viral↗
PubMed2000

Time-course of the auditory BOLD response to scanner noise.

It is a concern for auditory fMRI studies that acoustic noise generated by the scanner produces an auditory response that can confound stimulus-induced activation. To establish how to minimize this problem, the present study mapped the time-course of the auditory response to a burst of acoustic scanner noise by employing a single-event method. Recorded bursts of scanner noise were interspersed with clustered-volume acquisitions at a range of stimulus-to-imaging delays to map the response with a temporal resolution of 1 sec. There were strong responses (1.5% signal change) to scanner noise in primary and secondary auditory cortex. In both cortical areas, the mean response rose to a peak by 4-5 sec after stimulus onset and decayed after a further 5-8 sec. The time course indicates that noise contamination in auditory fMRI can be substantially reduced by using a 9-12-sec repetition time, thus maximizing the dynamic range available for displaying the response to acoustical stimuli of interest.

Adult↗
PubMed2006

Reducing protein concentration range of biological samples using solid-phase ligand libraries.

The discovery of specific polypeptides of diagnostic relevance from a biological liquid is complicated by the overall vast number and the large concentration range of all polypeptides/proteins in the sample. Depletion or fractionation methodologies have been used for selectively removing abundant proteins; however, they failed to significantly enrich trace proteins. Here we expand upon a new method that allows the reduction of the protein concentration range within a complex mixture, like neat serum, through the simultaneous dilution of high abundance proteins and the concentration of low abundance ones in a single, simple step. This methodology utilizes solid-phase ligand libraries of large diversity. With a controlled sample-to-ligand ratio it is possible to modulate the relative concentration of proteins such that a large number of peptides or proteins that are normally not detectable by classical analytical methods become, easily detectable. Application of this method for reducing the dynamic range of unfractionated serum is specifically described along with treatment of other biological extracts. Analytical surface enhanced laser desorption/ionization mass spectrometry (SELDI-MS) technology and mono- and two-dimensional electrophoresis (1-DE and 2-DE) demonstrate the increase in the number of proteins detected. Examples linking this approach with additional fractionation methods demonstrate a further increase in the number of detectable species using either the so-called "top down" or "bottom up" approaches for proteomics analysis. By enabling the detection of a greater proportion of polypeptides/proteins within a sample, this method may contribute significantly towards the discovery of new biomarkers of diagnostic relevance.

Chromatography, Ion Exchange↗
PubMed2026

Quantifying the aromatic amino acid metabolome: UPLC-MS/MS analysis of aromatic amino acids and their host and co-metabolites in plasma.

Aromatic amino acids (AAAs), tryptophan, phenylalanine, and tyrosine along with their pathway metabolites have been implicated in the pathogenesis of diseases ranging from cardiovascular, neurological, inflammatory, and cancer diseases, among others. As such, the measurement of the primary AAAs, their host pathway metabolites, and microbiome derived co-metabolites in blood can provide a sensitive reflection of systemic health. The aim of the study was to develop a method for the quantification of 17 metabolites, the three AAAs and various of their metabolites in plasma using a high-throughput ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method. The method demonstrated a dynamic range (1 to 16,700&#xa0;ng/mL), with detection limits (LOD) as low as 0.05&#xa0;ng/mL. Quantification limits ranged from 3 to 5019&#xa0;ng/mL (LLOQ) and up to 16,700&#xa0;ng/mL (ULOQ). Recovery at LQC, MQC, and HQC was satisfactory and consistent across most metabolites, with significant matrix effects observed only for 4-ethylphenol sulfate. Furthermore, intra and inter-day accuracy and precision met all acceptance criteria at all quality control concentrations for most of the metabolites. Measurement of NIST SRM 1950 showcased the method's accuracy for most of the metabolites. Finally, the method was applied on the analysis of plasma samples from 55 individuals (13 males and 42 females) providing information on AAAs and their pathway metabolites relevant concentrations in human plasma.

Amino Acids, Aromatic↗
PubMed2004

Development of a high-throughput plate-based chemiluminescent transcription factor assay.

Transcription factors are DNA-binding proteins that regulate the expression of specific genes by controlling transcription initiation. Two families of transcription factors, NFkappaB and AP-1, play pivotal roles in controlling important cellular processes ranging from normal cell growth and differentiation to apoptosis and cancer. Identifying changes in the DNA-binding activity of these factors is essential to understanding the regulation of these processes. We have developed a high-throughput DNA-based ELISA capable of monitoring activated levels of NFkappaB (p50 and p65) and AP-1 (c-Jun and c-Fos). This chemiluminescent assay utilizes a 96-well plate format, eliminating the throughput challenges imposed by traditional gel shift assays and exceeding the sensitivity and dynamic range of standard colorimetric detection systems. The sensitivity of this assay enables distinction between subtle as well as dramatic differences in the DNA-binding activity of these factors that result from the treatment of cells with various inhibitors or activating agents.

Animals↗
PubMed2006

Characterization of a second-generation focal-plane camera coupled to an inductively coupled plasma Mattauch-Herzog geometry mass spectrograph.

A second-generation Faraday-strip array detector has been coupled to an inductively coupled plasma Mattauch-Herzog geometry mass spectrograph, thereby offering simultaneous acquisition of a range of mass-to-charge ratios. The second-generation device incorporates narrower, more closely spaced collectors than the earlier system. Furthermore, the new camera can acquire signal on all collectors at a frequency greater than 2 kHz and has the ability to independently adjust the gain level of each collector. Each collector can also be reset independently. With these improvements, limits of detection in the hundreds of picograms per liter for metals in solution have been obtained. Some additional features, such as a broader linear dynamic range (over 7 orders of magnitude), greater resolving power (up to 600), and improved isotope ratio accuracy were attained. In addition, isotope ratio precision as low as 0.018% RSD was achieved.

Journal Article↗