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Biomedical subjects

W H Schuette

Publications and source records attributed to W H Schuette.

At least 19 recordsLinked to original sources

Slow oscillations of cytochrome oxidase redox state and blood volume in unanesthetized cat and rabbit cortex. Interhemispheric synchrony.

The purpose of this study was to determine the frequency characteristics and the degree of interhemispheric synchrony of slow (< 0.5 Hz), spontaneous oscillations of the regional cortical cytochrome oxidase redox state (CYT) and blood volume (CBV) in unanesthetized animals. We implanted bilateral cortical windows and electrodes for polysomnography in 7 cats and 3 rabbits. The animals were atraumatically restrained during multiple 3-6 hour sessions for up to 8 weeks, and relative changes in the cortical CYT and CBV were monitored by dual wavelength reflectance spectrophotometry at 603 nm and 590 nm. Continuous oscillations of CYT and CBV, unrelated to pulse or respiration, were always observed in each animal. Frequency (FFT) analysis over time revealed a nonstationary distribution of frequencies below 0.4 Hz, with most of the spectral power being contained in the 0-0.25 Hz band during both waking and sleep. Although the time-frequency plots of the CYT and CBV signals were similar, an occasional dissociation between the CYT and CBV oscillations was found. Analysis of simultaneous bilateral cortical optical recordings revealed a significant and sustained interhemispheric cross-correlation over time between the CYT as well as the CBV oscillations during stable recordings as long as 60 min. We conclude that: 1) CYT and CBV levels normally oscillate at < 0.4 Hz in the unanesthetized cat and rabbit cortex; 2) these complex oscillations, whose frequencies are non-stationary over time, nevertheless show sustained interhemispheric synchrony between 50 mm2 homotopic cortical regions; and 3) these oscillations may in part represent fluctuations of the metabolic rate.

Animals↗

Interhemispheric synchrony of slow oscillations of cortical blood volume and cytochrome aa3 redox state in unanesthetized rabbits.

In order to study spontaneous, slow oscillations of regional oxidative metabolism and blood flow in the normal, unanesthetized cortex, adult rabbits were implanted with bilateral cortical windows and electrodes for polysomnography. Relative changes in the cortical intramitochondrial redox state of cytochrome aa3 (CYT) and blood volume (CBV) were monitored by dual-wavelength reflectance spectrophotometry. Continuous, non-stationary oscillations (< 0.5 Hz) of both CYT and CBV were observed during waking and non-REM sleep. Cross-correlation analysis revealed a predominant interhemispheric synchrony of these oscillations which were unrelated to the heart rate, breathing, or electrocorticogram pattern. These findings suggest a dynamic linkage of slowly varying metabolic and vascular processes between unanesthetized cortical regions of 50 mm2 surface area.

Animals↗

Apparent unilateral visual neglect in MPTP-hemiparkinsonian monkeys is due to delayed initiation of motion.

Monkeys made hemiparkinsonian by infusion of a solution of MPTP into one carotid artery appeared to ignore food presented from the contralateral side. Initial observations suggested neglect of visual stimuli presented as fruit treats by automated delivery system in the half-field contralateral to MPTP treatment. Further studies in which fruit treats were left in the 'neglected' visual field indicated that this apparent neglect, unlike neglect attending cortical lesions, was rather a marked delay in initiating movements (unilateral hypokinesia). These observations may explain apparent subcortical neglect and are consistent with the known role of nigrostriatal dopaminergic neurones in movement regulation. This is a useful animal model in which difficulties in initiation of movement (hypokinesia). a cardinal symptom of Parkinson's disease, can be studied separately from other deficits in motor performance.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine↗

Low-frequency oscillations of cortical oxidative metabolism in waking and sleep.

To study the changes in cortical oxidative metabolism and blood volume during behavioral state transitions, we employed reflectance spectrophotometry of the cortical cytochrome c oxidase (cyt aa3) redox state and blood volume in unanesthetized cats implanted with bilateral cortical windows and EEG electrodes. Continuous oscillations in the redox state and blood volume (approximately 9/min) were observed during waking and sleep. These primarily metabolic oscillations of relatively high amplitude were usually synchronous in homotopic cortical areas, and persisted during barbiturate-induced electrocortical silence. Their mean amplitude and frequency did not vary across different behavioral/EEG states, although the mean levels of cyt aa3 oxidation and blood volume during rapid eye movement (REM) sleep significantly exceeded those during waking and slow-wave sleep. These data suggest the existence of a spontaneously oscillating metabolic phenomenon in cortex that is not directly related to neuroelectric activity. A superimposed increase in cortical oxidative metabolism and blood volume occurs during REM sleep. Experimental data concerning cerebral metabolism and blood flow that are obtained by clinical methods that employ relatively long sample acquisition times should therefore be interpreted with caution.

Animals↗

A simplified method for monitoring the cytochrome aa3 redox state in bilateral cortical areas of unanesthetized cats.

We describe a versatile optical system that enables the simultaneous monitoring of the redox state of cytochrome c oxidase (cytochrome aa3) in two homologous cortical areas under chronically implanted windows in cats. A single light source, broad bandpass primary filters, light-conducting rods, and narrow-bandpass interference detecting filters are employed. We observed reproducible responses of the cytochrome redox state and blood volume to carotid occlusion and terminal anoxia during anesthesia, and to graded doses of pentobarbital in awake animals.

Animals↗

Closed loop television tracking of beating heart cells in vitro.

The motion of beating heart cells in vitro has been used as a sensitive indicator of the presence of drugs. The combination of closed loop television video tracking and latex microspheres placed on the cells as markers results in an improved measurement of cell motion. Latex microspheres, 5.2 micrometers, are allowed to settle onto beating myocardial heart cells in vitro, where they move in concert with the cell motion. These microspheres, when viewed by televised transmission microscopy with normal optics, present high contrast circular targets that are excellent for closed loop television video tracking techniques. With video tracking, an analog voltage is generated proportional to the horizontal center of intensity of the target being tracked. This signal represents the location of a video demodulator gate that is maintained centered over the video target by feedback control.

Animals↗

An iterative method for the deconvolution of microcalorimeter thermograms.

An iterative method for the deconvolution of microcalorimetry thermograms suitable for small digital computers is presented. The method employs a measured impulse response function directly as the deconvolution kernel, thus explicit system simulation is not required. Data are presented showing the performance of the method and the exchange of signal-to-noise ratio for time resolution that is made when deconvolution techniques are employed. An improvement in the system time resolution of fifty times is demonstrated with measured data.

Algorithms↗

A system for measuring the angular response of hemiparkinsonian monkeys to a food stimulus.

A Zenith Z-100 PC system was used to control an automated feeding system for determining the angular limits at which a monkey would respond to a food stimulus. Raisins secured to the end of a rotating arm by means of vacuum pressure were delivered to the monkey alternately from clockwise and counterclockwise directions at a fixed radius. The point at which the monkey took a raisin from the end of the arm was determined by an increase in negative pressure. The arm position was measured by using a potentiometer mounted in the arm gear train. The computer system controlled the experiment as well as the on-line recording and plotting of data.

Analog-Digital Conversion↗

Count-dependent filter for smoothing bivariate FCM histograms.

A data-smoothing filter has been developed that permits the improvement in accuracy of individual elements of a bivariate flow cytometry (FCM) histogram by making use of data from adjacent elements, a knowledge of the two-dimensional measurement system point spread function (PSF), and the local count density. For FCM data, the PSF is assumed to be a set of two-dimensional Gaussian functions with a constant coefficient of variation for each axis. A set of space variant smoothing kernels are developed from the basic PSF by adjusting the orthogonal standard deviations of each Gaussian smoothing kernel according to the local count density. This adjustment in kernel size matches the degree of smoothing to the local reliability of the data. When the count density is high, a small kernel is sufficient. When the density is low, however, a broader kernel should be used. The local count density is taken from a region defined by the measurement PSF. The smoothing algorithm permits the reduction in statistical fluctuations present in bivariate FCM histograms due to the low count densities often encountered in some elements. This reduction in high-frequency spatial noise aids in the visual interpretation of the data. Additionally, by making more efficient use of smaller samples, systematic errors due to system drift may be minimized.

Cell Count↗

The relationship between mean channel selection and the calculated coefficient of variation.

Calculated coefficients of variation (CV) taken from the quotient of the standard deviation (S.D.) and the mean value of measured distributions are often used as an indicator of system performance in linear flow cytometry (FCM). The ability of the calculated CV to estimate the true CV of the underlying experiment before grouping (channelization) is dependent on the relationship between the width of the data channels and the magnitude of the S.D. of the measured distribution. When the channel width is equal to the S.D. of a distribution, the calculated CV is approximately 20% larger than the true CV of an experiment. By the time the S.D. is only one-half of a channel width, the calculated CV is unreliable. When the distribution S.D. is narrower than a channel's width, small changes in the distribution mean value will cause large variations in the calculated CV. As the true CV decreases, the calculation must be made with higher mean channel values. This dependence of calculated CV accuracy upon the relationship between S.D. and channel width places limitations upon mean channel selection that must be considered when using CV calculations for evaluating system performance, especially when looking for small improvements during optical alignment procedures. When an instrument is assumed to have a constant CV and the data are collected linearly, it is possible to improve the CV estimation accuracy by placing distributions in higher-numbered channels.(ABSTRACT TRUNCATED AT 250 WORDS)

Flow Cytometry↗

A count-dependent filter for smoothing flow cytometric histograms.

An adaptive count-dependent algorithm for smoothing statistically limited histograms has been developed. It considers both the spatial frequency limitations of the measurement system (described by the measurement system point spread function) and the reliability of the measured data (indicated by the effective number of counts influencing each channel of the histogram. Windows for smoothing flow cytometric histograms are derived from an assumed Gaussian-shaped point spread function (PSF) with a constant coefficient of variation. The windows are developed by scaling the variances of the Gaussian functions inversely with the statistical reliability of the data contained in each channel of the measured histogram. The reliability of this data is determined by taking the square root of the number of counts influencing the value tabulated for each channel. Using the algorithm, a smoothed version of the measured histogram may be developed from a linear sum of the products of the individual scaled Gaussian functions and the original measured histogram. Data are presented demonstrating the advantages of count-dependent smoothing over non-count-dependent smoothing using synthesized DNA histograms as a function of sample size.

Cell Cycle↗

Design of flow chamber with electronic cell volume capability and light detection optics for multilaser flow cytometry.

A multibeam optical detection system has been developed with a high optical efficiency, achieved through a reduction in the number of optical interfaces employed in the system. This reduction is made possible by a combination of employing simple lenses, gluing the objective lens directly upon the face of the flow cuvette and the extraction of only one fluorescence signal from each laser beam. A modified flow chamber is also described that includes fluidic resistance elements for the elimination of most of the electric shielding normally associated with electronic cell volume measurements.

Animals↗

The biology of tumor growth in the non-Hodgkin's lymphomas. A dual parameter flow cytometry study of 220 cases.

Dual parameter flow cytometry studies (cell DNA content and electronic cell volume) were performed in 220 cases of non-Hodgkin's lymphoma. All cases were characterized as B or T cell malignancies, based on immunologic surface marker characteristics. Aneuploidy by flow cytometry was more common among the B cell lymphomas than among the T cell lymphomas, and was most common among the large B cell lymphomas and B cell lymphomas of intermediate size. Ploidy index distributions showed a prominent hyperdiploid peak, as well as tumor cell populations with near-tetraploid DNA contents. In serial studies, a decrease in ploidy index was observed in association with clinical and histologic transformation in one case. The highest S fractions were observed among the large and intermediate B cell lymphomas and among the aggressive T cell lymphomas. In clinical samples consisting of mixtures of diploid and aneuploid populations, the data on the aneuploid components could often be separated from other components of the mixture in multiparameter studies on the basis of the larger electronic cell volumes of the aneuploid cells. In each case, the aneuploid large cell component almost invariably had a higher S fraction than the residual component(s) of the mixture. Overall, the data are consistent with a model of clonal selection and clonal evolution in the lymphomas in which early cytogenetic abnormalities that involve little or no change in total cell DNA content are followed by cell tetraploidization that is associated with cytogenetic instability and chromosome loss over the course of time.

Aneuploidy↗

High resolution method for the analysis of DNA histograms that is suitable for the detection of multiple aneuploid G1 peaks in clinical samples.

The DNA histogram obtained by flow cytometry can be considered as the product of an "ideal" measurement column vector and a measurement distortion matrix. In order to extract the ideal histogram from the real data, the measurement distortion matrix is commonly presumed to be a family of Gaussian coefficients that are centered on the diagonal. We have designed a feedback-controlled curve-fitting procedure that reconstructs the ideal histogram from the real data through successive iterations. The optimum coefficient of variation (cv) for the family of Gaussians in the measurement distortion matrix is determined from an analysis of the sums of squares of fits of the computed DNA histogram to the real data over an appropriate range of trial cv. Since this method assigns a Gaussian to each and every data channel, it permits the resolution of closely spaced multiple aneuploid G1 peaks in clinical samples. The effects of high frequency noise that may be present in the data can be attenuated by multiplying the real data histogram by a Gaussian matrix with cv close to but smaller than that of the measurement distortion matrix.

Aneuploidy↗

A practical graphical method for estimating the fraction of cells in S in DNA histograms from clinical tumor samples containing aneuploid cell populations.

A graphical method for the analysis of unperturbed DNA histograms is presented in which the area of the normalized histogram subtended by the fraction of cells in S is represented by a trapezoid whose dimensions are dependent on features common to all such histograms. The technique takes measurement variability into account. This method was applied to a variety of synthetic DNA histograms. Overall, calculated values for the fraction of cells in S correlated well with actual values. This method was applied to 36 diploid cases of non-Hodgkin lymphoma; results correlated well with those obtained by a computer-based method. The results of the graphical-method were also highly reproducible between different observers. The graphical method can be used in the presence of aneuploid cell populations. Techniques for calculating S fractions in the presence of aneuploidy in clinical samples are described. These techniques were applied to synthetic histograms of mixed diploid and aneuploid populations. Calculated values correlated well with actual values.

Aneuploidy↗

Automated radioautographic grain counting. Correction for grain overlap.

An algorithm is described for the calculation of radioautographic cell grain count from measurements of total cell nuclear area and total grain area. This algorithm provides a statistical correction for grain overlap that is based on the solution to the occupancy problem in probability theory. This method permits the use of automated grain counting over a wide range of grain counts per cell, and extends the useful dynamic range of radioautographic grain counting to well over 200 grains/cell.

Autoradiography↗

Effects of vincristine on cell survival, cell cycle progression, and mitotic accumulation in asynchronously growing Sarcoma 180 cells.

The effects of vincristine (VCR) on cell survival, cell cycle progression, DNA synthesis, and metaphase accumulation were studied in relation to drug concentration and drug exposure duration in Sarcoma 180 cells in vitro. VCR was found to affect cells in interphase, producing a transient G2 block at all drug concentrations and drug exposure durations studied. VCR did not affect DNA synthesis directly. Increases in the metaphase index were delayed and always peaked at approximately 8 hr after drug removal, regardless of the duration of drug exposure. Increases in the metaphase index of sufficient magnitude to be commensurate with VCR lethality were observed only with prolonged drug exposure. VCR produced both nuclear fragmentation and polyploidy. The proportion of cells undergoing polyploidy increased progressively with increasing drug exposure duration. Interference with cytokinesis during prolonged VCR exposure may represent a lethal effect of VCR that is separate from its short-term effects. This could serve as the basis for the clinical study of the antitumor effects of prolonged VCR infusions.

Animals↗