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PubMed1987

Binaural loudness summation in the hearing impaired.

Binaural loudness summation was measured using three different paradigms with 10 normally hearing and 20 bilaterally symmetrical high-frequency sensorineural hearing loss subjects. An adaptive paradigm and a loudness matching procedure measured summation at the lower and upper level of comfortable loudness and the loudness discomfort level (LDL). Monaural and binaural LDLs also were obtained with a clinical procedure designed to select maximum output of hearing aids. Stimuli for all three tasks consisted of 500- and 4000-Hz pure tones and a speech spectrum noise. Binaural summation increased with presentation level using the loudness matching procedure, with values in the 6-10 dB range. Summation decreased with level using the adaptive paradigm, and no summation was present with the clinical LDL task. The hearing-impaired subjects demonstrated binaural summation that was not significantly different from the normally hearing subjects. The results suggest that a bilaterally symmetrical sensorineural hearing loss does not affect binaural loudness summation. The monaural and binaural dynamic range widths were similar, and the LDL results suggest that binaural loudness summation may not be an important factor in selecting maximum output of hearing aids.

Acoustics↗
PubMed2004

The effects of high field strength MRI on electrodes and pulse generator in dynamic graciloplasty.

OBJECTIVE: Dynamic graciloplasty is a treatment for intractable faecal incontinence. A gracilis muscle is transposed around the anus and stimulated with an implanted pulse generator (IPG). This in vitro study was designed to determine the safety of MRI in patients with implanted electrodes and pulse generators for dynamic graciloplasty. METHODS: Temperatures were measured with fiberoptic probes around the devices in a cadaver model. Current was measured with an oscilloscope connected to electrodes and IPG. Movement and IPG parameter setting were observed before, during and after testing. RESULTS: Minor temperature increase under 1 degrees C were observed around the electrodes. Amplitudes measured were within the range of -1 and +1 Volt. No movement or changing of IPG parameters was noted. CONCLUSION: Changes noted are well within physiological ranges. Dynamic graciloplasty is not a contraindication for high field strength MRI.

Cadaver↗
PubMed2000

Assessment of viral loads in patients with chronic hepatitis C with AMPLICOR HCV MONITOR version 1.0, COBAS HCV MONITOR version 2.0, and QUANTIPLEX HCV RNA version 2.0 assays.

The correlation between response to antiviral therapy and pretreatment viral load in patients with chronic hepatitis C has prompted the development of quantitative assays to measure viral load. The aim of our study was to assess the clinical relevance of the newly developed semiautomated PCR system COBAS HCV MONITOR version 2.0 in comparison with (i) the AMPLICOR HCV MONITOR version 1.0 assay, which underestimates RNA concentration of hepatitis C virus (HCV) genotypes 2 to 6, and (ii) the QUANTIPLEX HCV RNA version 2.0 assay, which achieves equivalent quantification for each HCV genotype, with samples from 174 patients diagnosed with chronic hepatitis C before therapy. The level and range of quantification measured with AMPLICOR HCV MONITOR version 1.0 were 1 log lower than when measured with the COBAS HCV MONITOR version 2.0, at 0.261 x 10(6) RNA copies/ml (range, 0.001 x 10(6) to 2.50 x 10(6) RNA copies/ml) and 4.032 x 10(6) RNA copies/ml (range, 0.026 x 10(6) to 72.6 x 10(6) RNA copies/ml), respectively. The two assays showed a poor correlation (r(2) = 0.175). The level and range of quantification were similar when measured with the COBAS HCV MONITOR version 2.0 and QUANTIPLEX HCV RNA version 2.0 assays, at 3.03 x 10(6) RNA copies/ml (range, 0.023 x 10(6) to 72.6 x 10(6) RNA copies/ml) and 4.91 Meq/ml (range, 0.200 to 49.5 Meq/ml), respectively. The two assays showed a strong correlation (r(2) = 0. 686) for each HCV genotype. The duration of treatment (6 or 12 months) is modulated according to HCV genotype and viral load. Our results indicate that COBAS HCV MONITOR version 2.0 and QUANTIPLEX HCV RNA version 2.0 assays showing an equal dynamic range for each HCV genotype are suitable tools to assess patients before therapy.

Evaluation Studies as Topic↗
PubMed1976

Basis and some diagnostic implications of electrocochleography.

Electrocochleography (ECoG) involves the recording of electrical responses to sound from the vicinity of the cochlea. The technique, as we practice it, utilizes a trans-tympanic recording needle situated on the promontory of the middle ear. Filtered clicks in the frequency range between 500 and 8,000 Hz are presented to the ear under test at a rate of 10/sec. Repetitive clicks of a given frequency are first presented at high intensity and the responses summed (averaged) in a computer. The click is systematically lowered in intensity, and an average is collected at each level until the normal dynamic range of hearing has been explored. Two major electrical indices are present in the recording: these are 1. the whole-nerve action potential (AP) derived from the first-order auditory neurons, and 2. the cochlear microphonic (CM) derived from the hair cells. Inspection of the whole nerve AP as intensity is lowered allows the estimation of the response threshold, which correlates well with behavioral threshold. Inspection of the relation between the CM and the AP allows qualitative differentiation to be made between pathology arising in the hair cell (sensory) and in the nerve (neural).

Acoustic Stimulation↗
PubMed1994

Noise, amplification, and compression: considerations of three main issues in hearing aid design.

This paper deals with the following three topics: (1) interfering noise (voice babble, single competing speaker) as the main problem of many hearing-impaired listeners, (2) the amplitude-frequency response of the hearing aid, and (3) the benefit of frequency-dependent compression. Research by the author and his coworkers has shown that: (1) persons with impaired hearing typically need 3 to 6 dB higher speech-to-noise ratios than do normal-hearing listeners--a technically very difficult problem to solve; (2) within a relatively ample range, the speech-reception threshold in noise is independent of the amplitude-frequency response; and (3) the small time constants of syllabic compression deteriorate the speech signal. Multichannel amplification (2-4 channels) with automatic gain control for each channel is recommended, optimally adjusted to keep the (variable) speech signal within the impaired ear's limited dynamic range as well as to preserve the intensity differences of successive speech phonemes.

Acoustic Stimulation↗
PubMed2007

Mechanisms of DNA binding determined in optical tweezers experiments.

The last decade has seen rapid development in single molecule manipulation of RNA and DNA. Measuring the response force for a particular manipulation has allowed the free energies of various nucleic acid structures and configurations to be determined. Optical tweezers represent a class of single molecule experiments that allows the energies and structural dynamics of DNA to be probed up to and beyond the transition from the double helix to its melted single strands. These experiments are capable of high force resolution over a wide dynamic range. Additionally, these investigations may be compared with results obtained when the nucleic acids are in the presence of proteins or other binding ligands. These ligands may bind into the major or minor groove of the double helix, intercalate between bases or associate with an already melted single strand of DNA. By varying solution conditions and the pulling dynamics, energetic and dynamic information may be deduced about the mechanisms of binding to nucleic acids, providing insight into the function of proteins and the utility of drug treatments.

Animals↗
PubMed2004

Coding of acoustic particle motion by utricular fibers in the sleeper goby, Dormitator latifrons.

It is unknown whether the fish utricle contributes to directional hearing. Here, we report response properties of single utricular fibers in a teleost fish ( Dormitator latifrons) to linear accelerations at various stimulus frequencies and axes. Characteristic frequencies ranged from < or =50-400 Hz (median=80 Hz), and best frequencies shifted from 50 to 250 Hz with stimulus level. Best sensitivity of utricular fibers was distributed from -70 to -40 dB re: 1 g (mean=-52 dB), which is about 30 dB less sensitive than saccular fibers. Q50% fell between 0.16 and 11.50 (mean=2.04) at 15 dB above threshold. We observed temporal response patterns of entrained phase-locking, double phase-locking, phase-locked bursting, and non-phase-locked bursting. Most utricular fibers were directionally selective with various directional response profiles, and directional selectivity was stimulus-level dependent. Horizontal best-response axes were distributed in a 152 degrees range while mid-sagittal best-response axes were clustered around the fish longitudinal axis, which is consistent with the horizontal orientation of the utricle and morphological polarizations of utricular hair cells. Therefore, results of this study indicate that the utricle in this vertebrate plays an auditory role in azimuth and that utricular fibers extend the response dynamic range of this species in directional hearing.

Acceleration↗
PubMed2001

Quantitative detection of hepatitis B virus DNA in serum by a new rapid real-time fluorescence PCR assay.

A sensitive and accurate HBV DNA quantification assay is essential for monitoring hepatitis B virus (HBV) replication. This study evaluated a real-time PCR method performed in the LightCycler analyser for quantitative HBV DNA assay. HBV DNA results with this method were compared with those obtained using a branched-chain DNA (bDNA) solution hybridization assay. Real-time PCR was performed using two adjacent fluorescently labelled probes and primers corresponding to the HBV core gene. The same standard employed in the bDNA assay was used for calibration. Serum samples came from 193 HBV surface antigen (HBsAg)-positive patients (34 HBV e antigen (HBeAg)-positive and 93 with antibody to HBeAg (anti-HBe)), and 66 asymptomatic HBV carriers. In addition, we analysed serum samples from 8 anti-HBe-positive patients who had been receiving lamivudine treatment for more than three years. A linear standard curve was seen in the range from 10(3) to 10(8) copies/mL. In the reproducibility analysis, intra-assay coefficient of variation (CVs) at two known HBV DNA concentrations were 4% and 2% and interassay CVs were 6% and 4%. The median of serum HBV DNA by real-time PCR was 9.2 x 10(8) copies/mL in HBeAg-positive patients with persistently elevated alanine aminotransferase (ALT) levels, 1.3 x 10(7) copies/mL in anti-HBe-positive cases with persistently elevated ALT levels, 3.7 x 10(4) copies/mL in anti-HBe-positive patients with fluctuating ALT levels and 10(4) copies/mL in asymptomatic HBV carriers. The differences in HBV DNA levels among the various groups studied were statistically significant (P < 0.05). The cut-off between chronic hepatitis patients and asymptomatic carriers was found to be at a serum HBV DNA concentration of 5 x 10(4) copies/mL. Of the 109 serum samples with a viral load < 7.5 x 10(5) (negative by bDNA assay) 44 (40%) were positive by real-time PCR: 24 (56%) chronic hepatitis and 20 (33%) asymptomatic carriers. There was a positive association between HBV DNA levels determined by real-time PCR and ALT levels (P < 0.05), which was not observed with the bDNA assay for HBV DNA quantification. At 12 months of lamivudine treatment, 6 patients (75%) showed HBV DNA levels < 5 x 10(4) copies/mL (range < 10(3)-2 x 10(3)), significantly lower than at baseline. At 36 months, 2 of 8 (25%) showed HBV DNA levels persistently lower than 5 x 10(4) copies/mL (1.7 x 10(3), 6 x 10(3)). The LightCycler quantitative real-time PCR is a practical, sensitive, reproducible single-tube assay with a wide dynamic range of detection. The assay is automatic except for DNA extraction and the running time is only 70 min. The LightCycler real-time PCR is useful for identifying different states of HBV infection and for evaluating the efficacy of viral therapy.

DNA, Viral↗
PubMed2004

Guanylyl cyclase-activating proteins (GCAPs) are Ca2+/Mg2+ sensors: implications for photoreceptor guanylyl cyclase (RetGC) regulation in mammalian photoreceptors.

Guanylyl cyclase-activating proteins (GCAP) are EF-hand Ca(2+)-binding proteins that activate photoreceptor guanylyl cyclase (RetGC) in the absence of Ca(2+) and inhibit RetGC in a Ca(2+)-sensitive manner. The reported data for the RetGC inhibition by Ca(2+)/GCAPs in vitro are in disagreement with the free Ca(2+) levels found in mammalian photoreceptors (Woodruff, M. L., Sampath, A. P., Matthews, H. R., Krasnoperova, N. V., Lem, J., and Fain, G. L. (2002) J. Physiol. (Lond.) 542, 843-854). We have found that binding of Mg(2+) dramatically affects both Ca(2+)-dependent conformational changes in GCAP-1 and Ca(2+) sensitivity of RetGC regulation by GCAP-1 and GCAP-2. Lowering free Mg(2+) concentrations ([Mg](f)) from 5.0 mm to 0.5 mm decreases the free Ca(2+) concentration required for half-maximal inhibition of RetGC ([Ca]((1/2))) by recombinant GCAP-1 and GCAP-2 from 1.3 and 0.2 microm to 0.16 and 0.03 microm, respectively. A similar effect of Mg(2+) on Ca(2+) sensitivity of RetGC by endogenous GCAPs was observed in mouse retina. Analysis of the [Ca]((1/2)) changes as a function of [Mg](f) in mouse retina shows that the [Ca]((1/2)) becomes consistent with the range of 23-250 nm free Ca(2+) found in mouse photoreceptors only if the [Mg](f) in the photoreceptors is near 1 mm. Our data demonstrate that GCAPs are Ca(2+)/Mg(2+) sensor proteins. While Ca(2+) binding is essential for cyclase activation and inhibition, Mg(2+) binding to GCAPs is critical for setting the actual dynamic range of RetGC regulation by GCAPs at physiological levels of free Ca(2+).

Animals↗
PubMed2001

Real-time TaqMan PCR as a specific and more sensitive alternative to the branched-chain DNA assay for quantitation of simian immunodeficiency virus RNA.

We developed a rapid and highly reproducible assay based on real-time PCR (TaqMan, Applied Biosystems, Foster City, CA) to quantitate simian immunodeficiency virus (SIV) RNA in plasma samples. This assay was compared with the current branched-chain DNA assay (Bayer, Emeryville, CA). Results obtained with the real-time TaqMan PCR assay were comparable to those obtained with the branched-chain DNA assay in overlapping ranges of sensitivities (r = 0.9429, p < 0.05). However, the real-time TaqMan PCR assay was capable of detecting as few as 50 copies of RNA/ml, whereas branched-chain DNA was only sensitive to 1,500 copies of RNA/ml. Therefore, several animals that tested negative by branched-chain DNA were positive by realtime TaqMan PCR. Two false positive tests were also recorded for the branched-chain DNA test. False negative and positive tests were confirmed by cell culture isolation and conventional nested RT-PCR. The SIV TaqMan assay detected a wide range of wild-type, cloned, and recombinant SIV strains with similar amplification efficiency, including SIVmac251, SIVmac239, SIVmac239 containing the 184V mutation in RT, SIV1A11, SIVmac239 delta3, SIVmac-M4, and chimeras (SHIVs) containing specific HIV-1 genes, such as reverse transcriptase (RT-SHIV) or Env (SHIV-E). In conclusion, the high sensitivity, increased specificity, wide dynamic range, simplicity, and reproducibility of the real-time SIV RNA quantitation allow the screening of large numbers of samples and make this method especially suitable for measuring both viral DNA and RNA levels during vaccine and therapy studies.

Animals↗
PubMed1997

Evaluation of reduced solvent volume solid-phase extraction cartridges with analysis by gas chromatography-mass spectrometry for determination of 11-nor-9-carboxy-delta 9-THC in urine.

A sensitive and reliable method was developed for the identification and quantitation of 11-nor-9-carboxy-delta 9-tetrahydrocannabinol (THCCOOH) in urine using a microbed solid-phase extraction (SPE) column. Extensive method validation is presented that evaluates the recovery, linearity, precision, limit of quantitation, limit of detection, and capacity of THCCOOH in urine extracts using CLEAN SCREEN reduced solvent volume (RSV) SPE columns followed by gas chromatographic-mass spectrometric analysis. The mean recovery of THCCOOH at concentrations of 2, 3, 5, 15, 50, and 150 ng/mL was found to be 91% with coefficients of variation of 7.3% or less. Linearity of the method ranged from 1.95 to 1000 ng/mL with sensitivity at 1 ng/mL for THCCOOH. The sorbent was found to retain at least 1000 ng/mL of THCCOOH with no analyte breakthrough using the described method. Reduction in both processing times and total solvent volume is shown. The RSV SPE columns showed excellent efficiency and performance while demonstrating recoveries, cleanliness, and dynamic ranges comparable with standard SPE products.

Dronabinol↗
PubMed1983

Auditory intensity discrimination at high frequencies in the presence of noise.

Over a wide range of intensities, subjects were able to detect small differences in the intensity of a high-frequency band of noise that was presented with a relatively intense, complementary band-reject noise. This indicates that neither of two possible mechanisms for peripheral intensity coding, those based on timing and on spread of excitation, is necessary for the large dynamic range of human hearing. It is shown that the information available in the firing rate of a small number of nerve fibers can account for these data.

Auditory Threshold↗
PubMed2006

A sample preparation process for LC-MS/MS analysis of total protein drug concentrations in monkey plasma samples with antibody.

The determination of protein concentrations in plasma samples often provides essential information in biomedical research, clinical diagnostics, and pharmaceutical discovery and development. Binding assays such as ELISA determine meaningful free analyte concentrations by using specific antigen or antibody reagents. Concurrently, mass spectrometric technology is becoming a promising complementary method to traditional binding assays. Mass spectrometric assays generally provide measurements of the total protein analyte concentration. However, it was found that antibodies may bind strongly with the protein analyte such that total concentrations cannot be determined. Thus, a sample preparation process was developed which included a novel "denaturing" step to dissociate binding between antibodies and the protein analyte prior to solid phase extraction of plasma samples and LC-MS/MS analysis. In so doing, the total protein analyte concentrations can be obtained. This sample preparation process was further studied by LC-MS analysis with a full mass range scan. It was found that the protein of interest and other plasma peptides were pre-concentrated, while plasma albumin was depleted in the extracts. This capability of the sample preparation process could provide additional advantages in proteomic research for biomarker discovery and validation. The performance of the assay with the novel denaturing step was further evaluated. The linear dynamic range was between 100.9ng/mL and 53920.0ng/mL with a coefficient of determination (r(2)) ranging from 0.9979 and 0.9997. For LLOQ and ULOQ samples, the inter-assay CV was 12.6% and 2.7% and inter-assay mean accuracies were 103.7% and 99.5% of theoretical concentrations, respectively. For QC samples, the inter-assay CV was between 2.1% and 4.9%, and inter-assay mean accuracies were between 104.1% and 110.0% of theoretical concentrations.

Animals↗
PubMed2001

Quantitative assay for group M (subtype A-H) and group O HIV-1 RNA detection in plasma.

A quantitative HIV-1 test is described based on a competitive RT-PCR assay combined with a sandwich hybridization as a detection system. The internal RNA standard (IS) was designed specifically to be competitive during the amplification and during the hybridization step. Sample viral load determination was carried out with one RT-PCR in the presence of 10(3) IS copies. The HIV-1 copy number was calculated by reference to an external standard curve performed on known and increasing amounts of the reference HIV-1 (Ref HIV-1) RNA co-amplified with a constant amount of the IS RNA. The assay had a linear range from 10(1) to 10(6) HIV-1 copies. HIV-1 strains belonging to the different subtypes from group M, but also group O, were all detected. Absolute quantification of purified HIV-1 RNA copies gave identical results as the AMPLICOR HIV-1 Monitor assay. The quantification of patient's samples was evaluated according to different criteria such as dynamic range, sensitivity, efficacy of material recovery, reproducibility and convenience of sample handling. The microplate format of the assay combined with the colorimetric detection provides a convenient tool and fulfills the requirement for routine molecular diagnostic laboratories.

HIV Infections↗
PubMedDate not supplied

Use of reporter genes for optical measurements of neoplastic disease in vivo.

Revealing the cellular and molecular changes associated with cancer, as they occur in intact living animal models of human neoplastic disease, holds tremendous potential for understanding disease mechanisms and elucidating effective therapies. Since light is transmitted through mammalian tissues, at a low level, optical signatures conferred on tumor cells by expression of reporter genes encoding bioluminescent and fluorescent proteins can be detected externally using sensitive photon detection systems. Expression of reporter genes, such as the bioluminescent enzyme firefly luciferase (Luc) or variants of green fluorescent protein (GFP) in transformed cells, can effectively be used to reveal molecular and cellular features of neoplasia in vivo. Tumor cell growth and regression in response to various therapies have been evaluated non-invasively in living experimental animals using these reporter genes. Detection of Luc-labeled cells in vivo was extremely sensitive with signals over background from as few as 1000 human tumor cells distributed throughout the peritoneal cavity of a mouse with linear relationships between cell number and signal intensity over five logs. GFP offers the strength of high-resolution ex vivo analyses following in vivo localization of the tumor. The dynamic range of Luc detection allows the full disease course to be monitored since disease progression from small numbers of cells to extensive disease can be assessed. As such, therapies that target minimal disease as well as those designed for late stage disease can be readily evaluated in animal models. Real time spatiotemporal analyses of tumor cell growth can reveal the dynamics of neoplastic disease, and facilitate rapid optimization of effective treatment regimens. Thus, these methods improve the predictability of animal models of human disease as study groups can be followed over time, and can accelerate the development of therapeutic strategies.

Animals↗
PubMed1980

High frequency Bekesy audiometry. I: equipment and calibration.

A simple, straightforward and inexpensive conversion of the high-frequency (HF) range (up to 20 kc/s) of the Grason-Stadler E-800 Bekesy audiometer is described, incorporating as transducer a 1/2-inch condenser microphone loosely coupled to the ear canal by an adjustable headband, which provides also for a standard audiometric earphone-cushion unit on the non-test ear. A free field calibration is recommended. The system has a dynamic range of 135 db. Step-by-step instructions are provided for the conversion and for calibrations. Repeated calibrations of a prototype unit showed acceptable stability and validity. The HF E-800 audiometer could provide a valuable and cost-effective source of HF diagnostic information for the audiologist.

Audiometry↗
PubMed1995

Perceptibility of defects in an aluminum test object: a comparison of the RVG-S and first generation VIXA systems with and without added niobium filtration.

OBJECTIVE: To compare the first generation VIXA (Gendex, Milan, Italy) and the RVG-S (Trophy Radiologie, Vincennes, France) for detection of defects in an aluminium test object at various exposures both with and without added niobium filtration. METHODS: Images of a 7 mm aluminium test object with defects ranging from 0.1 to 1.5 mm with standardized projection geometry. Seven dentists acted as observers. Perceptibility curves were developed for both sensors under the various filtration conditions and exposures. RESULTS: At optimum exposures both the VIXA and the RVG-S permitted the same number of defects to be observed. The optimum exposures were similar for the two systems, but the dynamic range was greater for the RVG-S. Addition of niobium filtration did not appreciably alter the radiation dose required to perceive a given number of defects using either system. CONCLUSIONS: The two systems performed equally at optimum exposure when the dose is approximately the same irrespective of the presence or absence of niobium filtration. Beyond the optimal level, the RVG-S outperformed the VIXA system.

Aluminum↗
PubMed2004

Application of oxidants to the spectrophotometric determination of amlodipine besylate in pharmaceutical formulations.

Three new spectrophotometric methods for the determination of amlodipine besylate have been proposed. The first two methods, i.e. A and B, are based on the oxidation of the drug with Fe(III) and the estimation of Fe(II) produced after chelation with either 1,10-phenanthroline or 2,2'-bipyridyl at 500 and 515 nm, respectively. The Beer's law was obeyed in the concentration ranges of 2-10 and 4-14 microg ml(-1) with molar absorptivity of 2.9 x 10(4) and 2.7 x 10(4) l mol(-1) cm(-1) for methods A and B, respectively. The third procedure depends on the interaction of amlodipine besylate with ammonium heptamolybdate tetrahydrate, which resulted in the formation of molybdenum blue (lambda(max) 825 nm). The linear dynamic range and the molar absorptivity values were found to be 15-59 microg ml(-1) and 1.8 x 10(4) l mol(-1) cm(-1), respectively. The results of the proposed procedures were validated statistically and compared with those obtained by the reference method. The proposed methods were applied successfully to the determination of amlodipine besylate in commercial tablets.

Amlodipine↗