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An active process in cochlear mechanics.

A model for cochlear mechanics is proposed to take account of its two systems, one passive and one active. The classical passive system stimulates the inner hair cells directly at levels above about 40 dB SL. At intensities below about 60 dB an active process, the 'cochlear amplifier' (CA), somehow provides additional energy that enhances the vibration of a narrow segment of the basilar membrane near the apical foot of the familiar, traveling wave envelope. The outer hair cells are essential for CA. The active system acts like a high-Q acoustic resonator, and it accounts for the great sensitivity and sharp tuning expressed by the 'tips' of neural tuning curves. The tips are selectively vulnerable to anoxia, noise exposure and other trauma. The CA model explains the detection of small differences in time as well as in frequency, the dual character of the electrocochleogram, recruitment of loudness in cochlear hearing impairment, the long latency of normal neural responses near threshold, acoustic emissions (both stimulated and spontaneous) and the locus of TTS in the frequency range above the exposure tone. Both the classical high-intensity system and the active low-level CA system are highly nonlinear and they combine to compress the great dynamic range of hearing into a much narrower range of mechanical movement of the cilia of the inner hair cells. The mechanism of CA is unknown, and the problem remains of how its action can be triggered by submolecular movements near threshold.

Auditory Threshold↗

Evaluation of a commercially available human serum amyloid A (SAA) turbidometric immunoassay for determination of equine SAA concentrations.

The aim of the present study was to evaluate whether equine serum amyloid A (SAA) concentrations could be measured reliably with a turbidometric immunoassay (TIA) developed for use with human serum. Intra- and inter-assay imprecision were evaluated by multiple measurements on equine serum pools. Assay inaccuracy was determined by linearity under dilution. The assay was subsequently used for measuring SAA concentrations in clinically healthy horses, horses with inflammatory diseases, horses with non-inflammatory diseases, and in horses before and after castration. In pools with low, intermediate and high SAA concentrations, the intra-assay imprecisions were 24.4%, 1.6% and 2.1%, and the inter-assay imprecisions were 33.2%, 4.6% and 6.5%. Slight signs of inaccuracy were observed, but these inaccuracies were negligible when considering the large dynamic range of the SAA response. The assay was able to detect the expected difference in SAA levels in different groups of horses. It was also able to demonstrate the expected dynamic changes in SAA after castration. In conclusion, equine SAA concentrations can be measured reliably using the TIA designed for human SAA.

Animals↗

Vagal, sympathetic and somatic sensory inputs to upper cervical (C1-C3) spinothalamic tract neurons in monkeys.

1. Myocardial ischemia activates vagal and sympathetic cardiac afferent fibers. The purpose of this study was to determine a neuro physiological basis for cardiac pain referred to C1-C3 somatic dermatomes. We hypothesized that afferent fibers traveling in vagal or sympathetic nerves transmit nociceptive information to C1-C3 spinothalamic tract (STT) neurons. 2. Electrical stimulation of the left stellate ganglion to excite cardiopulmonary sympathetic afferent fibers increased extracellular activity of 44 of 77 C1-C3 STT neurons examined in 33 anesthetized male monkeys (Macaca fascicularis); responses increased as stimulus strength increased. Additionally, this stimulus inhibited 5 cells, increased/decreased activity of 2 cells, and did not affect 26 cells. 3. Electrical stimulation of the left (ipsilateral) thoracic vagus nerve excited 41 of 78 C1-C3 STT neurons, inhibited 4 neurons, increased/decreased activity of 2 neurons, and did not affect 31 neurons. Responses increased with increasing stimulus strength Contralateral vagal stimulation excited 7 of 39 cells tested, inhibited 4 cells and did not affect 28 cells. 4. Effects of stimulating one or more vagal branches were examined on 22 C1-C3 STT neurons excited by input from left thoracic vagus nerve. Stimulation of the cardiac branch excited 11 of 16 cells tested; stimulation of the recurrent laryngeal nerve excited 11 of 18 cells; stimulation of vagal fibers just rostral to the diaphragm excited 8 of 19 cells. 5. Excitatory somatic receptive fields ranged from small ipsilateral fields to large, sometimes bilateral or noncontinuous fields. Many fields included the ipsilateral neck and/or inferior jaw. Thirty-nine of 74 neurons examined were wide dynamic range (WDR), 21 were high threshold (HT), 6 were low threshold (LT), and 8 did not respond to brushing or noxious pinching of somatic tissues. Most (38 of 39) WDR cells responded to stimulation of the stellate ganglion or vagal fibers, as did 18 of 21 HT cells, 3 of 6 LT cells, and 2 of 8 cells unresponsive to brush or pinch stimuli. 6. Results of this study supported the concept that vagal and/ or sympathetic afferent activation of C1-C3 STT neurons might provide a neural mechanism for referred pain that originates in the heart or other visceral organs but is perceived in the neck and jaw region. Additionally, C1-C3 STT neurons processed sensory information from widespread regions of the body.

Adrenergic Fibers↗

Dynamic systemic vascular resistance in a sheep supported with a Nimbus AxiPump.

Changes in systemic vascular resistance (SVR) in response to diminished pulse perfusion were analyzed over a dynamic range of flow conditions. An axial flow LVAD (Nimbus AxiPump, Rancho Cordova, CA) was implanted in a sheep for 28 days, during which time SVR was determined over several conditions of posture and excitability. Total arterial resistance (TR) was calculated dynamically as an index of SVR by analysis of pump flow in diastole, and systemic pressure estimated from the characteristic pressure-flow-speed relation of the AxiPump. TR was evaluated over a range of flow rates, including maximum flow--for which the pressures and flows were essentially nonpulsatile. Throughout the course of support, and independent of pulsatility, TR dropped when the sheep stood and was significantly lower than that in the sitting position (P < 0.01). Response to excitement followed the same trend: TR was significantly higher during agitation than during normal temper (P < 0.01). In spite of changes in pulse pressure and flow rate, SVR changes occurred according to expected physiologic responses for pulsatile perfusion. Because pump flow and pressure are sensitive to afterload, the results of these studies suggest that pump speed control must compensate for changes in SVR to maintain acceptable perfusion.

Animals↗

Imaging and quantitation of chemiluminescence using photoexcitable storage phosphor screen.

A novel storage phosphor (SrS:Ce,Sm) imaging screen optimized for the detection of luminescence has been developed. When coupled with an appropriate scanning system, this screen exhibits the following characteristics: (i) a range of visible light detection between 400 and 500 nm with a peak response at 450 nm; (ii) sensitivity comparable to X-ray film; (iii) 200-microns resolution; (iv) linear dynamic range greater than four orders of magnitude; and (v) a uniformity of response of +/- 1.32% coefficient of variation. These features facilitate the detection and accurate quantitation of light radiation produced from the enzymatic catalysis of chemiluminescent substrates such as 1,2-dioxetanes and luminol. Its suitability for chemiluminescent applications is demonstrated by the analysis of cosmid hybridization and enzyme-linked immunosorbent assay samples.

Alkaline Phosphatase↗

Perception of linear horizontal self-motion induced by peripheral vision (linearvection) basic characteristics and visual-vestibular interactions.

The basic characteristics of the sensation of linear horizontal motion have been studied. Objective linear motion was induced by means of a moving cart. Visually induced linear motion perception (linearvection) was obtained by projection of moving images at the periphery of the visual field. Image velocity and luminance thresholds for the appearance of linearvection have been measured and are in the range of those for image motion detection (without sensation of self motion) by the visual system. Latencies of onset are around 1 sec and short term adaptation has been shown. The dynamic range of the visual analyser as judged by frequency analysis is lower than for the vestibular analyser. Conflicting situations in which visual cues contradict vestibular and other proprioceptive cues show, in the case of linearvection a dominance of vision which supports the idea of an essential although not independent role of vision in self motion perception.

Adaptation, Psychological↗

The Tinsley LCR Databridge Model 6401 and electrical impedance measurements to evaluate skin integrity in vitro.

Electrical impedance is used to confirm skin integrity for in vitro dermal regulatory testing and as a tool to evaluate skin condition to determine the irritation and corrosion potential of various chemicals and personal care products. In this experiment, samples of dermatomed human skin were mounted onto static diffusion cells (0.64 cm2) maintained at 32 degrees C. Following equilibration with 0.9% saline in the donor and receptor chambers, an impedance measurement was taken with a Tinsley LCR Databridge Model 6401 set in the resistance mode (R) and in (a) the serial-equivalent mode (SER) with an alternating current (AC) frequency of 100 hertz (Hz), (b) SER and 1000 Hz, (c) parallel-equivalent mode (PAR) and 100 Hz, and (d) PAR and 1000 Hz. With the databridge set in the SER-equivalent mode and an AC frequency of 1000 Hz, the minimum (7.2 kOmega), maximum (10.0 kOmega), and median (8.6 kOmega) impedance values exhibited a limited response range (2.8 kOmega). However, when the Tinsley 6401 was set in the PAR-equivalent mode at the lower AC frequency of 100 Hz the minimum (16.7 kOmega), maximum (134.6 kOmega), median (83.2 kOmega), and range (117.9 kOmega) of values were the highest obtained. The results confirm that the operator-selected settings on the Tinsley LCR Databridge Model 6401 affect the impedance measurement and the dynamic range of values observed for dermatomed human skin in vitro.

Cadaver↗

Reliability of the Cambridge Low Contrast Gratings.

We examined the reliability of the Cambridge Low Contrast Gratings over four sessions at intervals of 1 hour, 1 day and 1 week. The 95% ranges of difference scores were found to be +/- 10.3 and +/- 10.5 test units for left and right eyes respectively. These reliability estimates span almost one-quarter of the test's dynamic range and one-third of the subjects' performance range. Practice effects were not found to be significant. Though previous reports have emphasized the usefulness of the Cambridge Gratings in screening for eye disease, caution is advised when interpreting changes that occur over time.

Adolescent↗

Intensity discrimination, increment detection, and magnitude estimation for 1-kHz tones.

Intensity difference limens (DLs) were measured over a wide intensity range for 200-ms, 1-kHz gated tones and for 200-ms increments in continuous 1-kHz tones. Magnitude estimates also were obtained for the gated tones over a comparable intensity range. The discrimination data are in general agreement with those from earlier studies but they extend them by showing: (1) good discrimination for gated tones over at least a 115-dB dynamic range; (2) a slight increase in the relative DL (delta I/I) as intensity increases above 95 dB SPL; (3) smaller DLs for increments than for gated tones, with the difference approximately independent of intensity; (4) negligible "negative masking" when thresholds are expressed as intensity differences (delta I). For two of the three subjects, magnitude estimates do not conform to a single-exponent power law for suprathreshold intensities. Over the middle range of intensities where a single exponent is appropriate, the value of the exponent is less than 0.1 for all subjects.

Acoustic Stimulation↗

Multicenter performance evaluation of a new TaqMan PCR assay for monitoring human immunodeficiency virus RNA load.

A TaqMan real-time PCR assay, the COBAS TaqMan human immunodeficiency virus (HIV) (HPS-CTMHIV) PCR assay, recently developed for the quantification of HIV type 1 RNA in plasma, was evaluated in comparison with the licensed COBAS AMPLICOR HIV-1 MONITOR (CAHIM) assay. In this study, we have analyzed the tests' sensitivities, precisions, and linearities using multiple replicates of a standard panel of HIV RNA covering a 7-logarithm range of concentrations, as well as serial threefold dilutions of high-titer clinical samples. The subtype inclusivity was also investigated, using a panel of subtypes A to H, while a collection of 160 clinical samples was analyzed to assess the tests' specificities and the systems' similarities. The results of these experiments showed that the HPS-CTMHIV assay has a sensitivity of 53 copies/ml (95% hit rate), 100% specificity, and good intra- and interassay precision. The results of the HPS-CTMHIV assay were linear in the 50- to 10(7)-copies/ml range, with a correlation coefficient (R) for expected versus observed results of 0.98. Compared to the CAHIM assay, the HPS-CTMHIV assay showed a high correlation (R=0.99) across the dynamic range of RNA concentrations that, for the CAHIM assay, requires two different sample preparations. Equivalent performances were also observed for the two systems in the detection and quantification of HIV subtypes A to H. These data indicate that the HPS-CTMHIV assay may be one of the tests of choice for monitoring viral load throughout the course of HIV infection and during highly active antiretroviral therapy.

Analysis of Variance↗

A review of nanoelectrospray ionization applications for drug metabolism and pharmacokinetics.

Although traditionally reserved for proteomic analysis, nanoESI has found increased use for small molecule applications related to drug metabolism/pharmacokinetics (DMPK). NanoESI, which refers to ESI performed at flow rates in the range of 200 to 1000 nL/min using smaller diameter emitters (10 to 100 microm id), produces smaller droplets than conventional ESI resulting in more efficient ionization. Benefits include greater sensitivity, enhanced dynamic range, and a reduced competition for ionization. These advantages may now be harnessed largely due to the introduction of a commercial system for automated nanoESI infusion. This development in turn has allowed ADME (absorption, distribution, metabolism, and excretion) scientists to consider novel approaches to mass spectrometric analysis without direct LC interfacing. While it is freely acknowledged that nanoESI infusion is not likely to supplant LC-MS as the primary analytical platform for ADME, nanoESI infusion has been successfully applied to both quantitative (bioanalysis) and qualitative (metabolite identification) applications. This review summarizes published applications of this technology and offers a perspective on where it fits best into the DMPK laboratory.

Animals↗

Radioimmunohistochemistry of epidermal growth factor receptor in breast cancer.

CONTEXT: Conflicting reports of epidermal growth factor receptor (EGFR) expression in breast cancer and inconstant relationships with established prognostic indicators and outcomes suggest difficulties with EGFR measurement. OBJECTIVE: To compare EGFR measurement in a panel of cell lines and in breast carcinomas by radioimmunohistochemistry (R-IHC), conventional immunohistochemistry (IHC), and a ligand binding (LB) assay. DESIGN: Eight EGFR-expressing cell lines and 50 primary breast carcinoma specimens were analyzed for EGFR by IHC, R-IHC, and LB assays. A further 153 primary breast cancer specimens were analyzed by R-IHC alone. RESULTS: All 3 assays were in good agreement for the cell lines. In the subset of the 50 carcinoma specimens, EGFR was detected by LB assays in 19 (38%) and by IHC in 24 (48%). However, R-IHC detected EGFR in 46 (92%) of 50 and in 186 (92%) of all 203 carcinoma specimens. The LB assay agreed poorly with R-IHC of carcinomas, possibly because the LB assay is sensitive to EGFR-expressing nontumor breast parenchyma in the tissue analyzed. Both IHC and R-IHC on carcinoma specimens agreed better, but 26 carcinoma specimens (52%) in which EGFR was not detectable by IHC had a 10-fold range in receptor level detectable by R-IHC. CONCLUSION: To elucidate the role of EGFR or other growth factor receptors in breast cancer requires accurate, sensitive receptor assays. With its dynamic range, R-IHC returned meaningful results over the entire range of expression actually present in breast cancer, which LB assays and IHC failed to do.

Adult↗

Auditory brainstem response to clicks in quiet, notch noise, and highpass noise.

Auditory brainstem responses to clicks in quiet, notch noise, and highpass noise were recorded from 10 normal-hearing adults. Contrary to some reports, the latency of wave V increased as the center frequency of the notch decreased. Response identifiability and wave V amplitudes were similar for the notch and highpass noises. Thus, the additional frequencies below the notch did not contribute sufficiently to the response to alter identifiability or amplitude. Notch noise, as compared to the highpass noise, is advantageous because of the increased frequency-specificity provided by the low-frequency masking noise. Presenting clicks in highpass or notch noise centered at 500 Hz is of limited value for assessing auditory sensitivity. This is because the range of testable hearing levels is restricted by (1) a high normal ABR threshold (approximately 65 dB nHL) and (2) the high noise levels required to mask the click. This limited dynamic range for assessing hearing loss is a major limitation of click in noise testing, particularly for 500 Hz. Generating a two-point audiogram by presenting clicks in noise with notches centered around 1000 Hz and 4000 Hz may prove promising for assessing auditory sensitivity.

Acoustic Stimulation↗

Temporal patterning in the responses of gracile and cuneate neurones in the cat to cutaneous vibration.

1. Recordings were made in decerebrate cats from gracile and cuneate neurones responding to vibration-induced inputs from Pacinian corpuscle (P.c.) receptors of the hind-limb and forelimb footpads. The two groups of neurones were compared, in particular for their capacities for responding to cutaneous vibration with phase-locked impulse patterns. 2. In both nuclei the P.c. neurones were most sensitive to vibration in the range 80 to greater than 600 Hz. Stimulus-response relations were similar for the two groups, as were measures derived from these relations such as response levels, absolute thresholds and the dynamic range (defined as the vibration amplitude range over which responses were graded). 3. At frequencies up to 300-400 Hz, responses for some neurones in both nuclei remained well phase locked to the vibration; however, quantitative analysis using a factorial analysis of variance indicated that the phase locking was poorer in gracile than cuneate neurones. 4. In both nuclei there was marked variability from neurone to neurone in measures of phase locking which may reflect variations in the extent of convergence of P.c. fibres upon different target neurones. For neurones in either nucleus that had comparatively tight phase locking of responses to vibration it is proposed that their output is functionally dominated by one or a few of their convergent P.c. input fibres.

Action Potentials↗

Polariscope for simultaneous measurement of the principal axis and the phase retardation by use of two phase-locked extractions.

A novel polariscope with electro-optic modulation that is capable of simultaneous measurement of the principal axis and the phase retardation of an optical linear birefringent medium by means of two phase-locked extractions is described. A phase compensator is used to suppress the transmission phase-retardation effect of the beam splitter, thereby enhancing the precision of the measuring performance. The validity of the proposed design is demonstrated by measurement of the principal axis and phase retardation of a quarter-wave plate sample. There are absolute errors of 0.25 degrees on average and 0.58 degrees at maximum in the principal-axis measurement and of 0.75 degrees (0.83%) on average and 3.11 degrees at maximum in the phase-retardation measurement. Meanwhile, the retardation error lies within a 5% uncertainty range of a commercial wave plate. The root-mean-square resolutions for the principal-axis angle and phase-retardation measurements are 0.042 degrees and 0.081 degrees, respectively. Finally, the dynamic ranges of the principal-axis angle measurement and the phase-retardation measurement extend as far as 180 degrees.

Journal Article↗

Photodynamic actinometry using microspheres: concept, development and responsivity.

Photodynamic therapy (PDT) relies on three main ingredients, oxygen, light and photoactivating compounds, although the PDT response is definitively contingent on the site and level of reactive oxygen species (ROS) generation. This study describes the development of a novel, fluorescent-based actinometer microsphere system as a means of discerning spatially resolved dosimetry of total fluence and ROS production. Providing a high resolution, localized, in situ measurement of fluence and ROS generation is critical for developing in vivo PDT protocols. Alginate-poly-L-lysine-alginate microspheres were produced using ionotropic gelation of sodium alginate droplets, ranging from 80 to 200 microm in diameter, incorporating two dyes, ADS680WS (ADS) and Rhodophyta-phycoerythrin (RPE), attached to the spheres' inside and outside layers, respectively. To test the responsivity and dynamic range of RPE for ROS detection, the production of ROS was initiated either chemically using increasing concentrations of potassium perchromate or photochemically using aluminum tetrasulphonated phthalocyanine. The generation of singlet oxygen was confirmed by phosphorescence at 1270 nm. The resulting photodegradation and decrease in fluorescence of RPE was found to correlate with increased perchromate or PDT treatment fluence, respectively. This effect was independent of pH (6.5-8) and could be inhibited using sodium azide. RPE was not susceptible to photobleaching with light alone (670 nm; 150 Jcm(-2)). ADS, which absorbs light between 600 and 750 nm, showed a direct correlation between radiant exposure (670 nm; 0-100 Jcm(-2)) and diminished fluorescence. Photobleaching was independent of irradiance (10-40 mW cm(-2)). We propose that actinometer microspheres may provide a means for obtaining high spatial resolution information regarding delivered PDT dose within model systems during investigational PDT development and dosimetric information for clinical extracorporeal PDT as in the case of ex vivo bone marrow purging.

Bone Marrow Purging↗

Chromatographic competitive binding immunoassays: a comparison of the sequential and simultaneous injection methods.

Two approaches for performing competitive binding immunoassays by HPLC and other flow-based systems are the simultaneous and sequential injection methods. Both these techniques make use of a column with a limited amount of antibody, onto which is injected a sample and a fixed amount of a labeled analyte analog. An indirect measure of the unlabeled analyte in the sample is then obtained by looking at the amount of analog in either the nonretained or retained peaks. In the simultaneous injection mode, the sample and labeled analog are applied at the same time to the column, while in the sequential mode the sample is injected first, followed by the analog. This results in a difference in the analytical characteristics of these two approaches. This study used chromatographic theory and previous data obtained for injections of human serum albumin (HSA) onto an anti-HSA antibody column to compare the response, detection limits, range, and sensitivity of these methods. Under equivalent conditions, it was found that the sequential method always provided the best lower limit of detection and sensitivity. However, the simultaneous mode had a broader dynamic range and higher upper limit of detection. From these observations, several guidelines were developed regarding the use and selection of such assays for new applications.

Binding, Competitive↗

Low peripheral blood viral HIV-2 RNA in individuals with high CD4 percentage differentiates HIV-2 from HIV-1 infection.

OBJECTIVES: To elucidate why the virulence of HIV-1 and HIV-2 infections differ in West African populations. STUDY DESIGN/METHOD: Peripheral blood plasma virion RNA and cellular proviral DNA levels were measured in a cross-section of 59 HIV-1 and 49 HIV-2 singly infected individuals representing all stages of infection in The Gambia, West Africa. Novel reverse transcriptase polymerase chain reaction (RT-PCR) assays specific and sensitive for virus quantification of non-clade B HIV-1 and HIV-2 infections were used. RESULTS: HIV-1 and HIV-2 proviral and plasma RNA levels were inversely correlated with CD4+ count for both infections with cellular proviral load similar at each stage of infection. Critically, up to three-fourths of HIV-2-infected individuals with high CD4 percentages (> 28%) had undetectable (< 500 copies/mL) levels of peripheral blood HIV-2 RNA in contrast to HIV-1-infected individuals who had readily detectable plasma virus at all stages of infection (P < .0001). Plasma RNA levels were similar in the intermediate and end stages of infection, indicating similar replication potential for both viruses. In the cross-section of HIV-1- and HIV-2-infected patients studied, the data indicate a wider dynamic range of HIV-2 RNA in vivo compared with HIV-1. DISCUSSION: Low levels of HIV-2 replication and virion expression characterize individuals with high CD4+ lymphocyte counts, suggesting that a very different dynamic equilibrium exists between virus and host for HIV-2 compared with HIV-1. By analogy with HIV-1, our data implicate a considerably lower turnover of HIV-2 virion RNA in vivo with a markedly reduced production of infectious genomes in individuals during the subclinical phase of infection. CONCLUSION: The lower levels of virion expression of HIV-2 infections in vivo are compatible with observed differences in the natural history of HIV-1 and HIV-2 infections, relating to overall differences in the pathogenesis and disease progression of the two infections.

Acquired Immunodeficiency Syndrome↗