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Preload response in patients after cardiac surgery: a comparison of systolic blood pressure and systolic area variability and initial volume of distribution of glucose.

OBJECTIVE: Estimation of stroke volume variation (e.g. systolic blood pressure and systolic area variability) and central extracellular compartment volume (e.g. initial volume of distribution of glucose, IVDG) may be useful in guiding fluid therapy in mechanically ventilated patients. The reliability of systolic blood pressure (SBP) variability has been well validated, but little is known about systolic area (SA) variability or IVDG. Our aim was to investigate SBP and SA variability and IVDG as predictors of preload responsive hypovolaemia in post-cardiac surgery patients. METHODS: Thirty-four mechanically ventilated patients undergoing preload enhancement post elective cardiac surgery were studied. The maximum-minimum difference and power spectral measurement of SBP and SA variability were derived from the arterial waveform trace and examined before and after rapid volume infusion. IVDG was determined prior to volume infusion from three-minute incremental glucose estimation and (with SBP and SA variability) correlated with subjects classified as hypotensive and preload responsive. RESULTS: Neither IVDG or SA and SBP variability were found to correlate with subjects identified as hypotensive and preload responsive. However, the power spectral measures of SBP and SA variability were significantly reduced (p = 0.007 and p = 0.026, respectively) following preload enhancement in fluid responsive subjects. CONCLUSIONS: Our results indicate that neither IVDG, nor SBP and SA variability are predictive of preload responsive hypotension in post-cardiac surgery patients. Spectral analysis of SBP and SA may be more sensitive at assessing preload responsiveness in this patient group than traditional maximum-minimum measures.

Journal Article↗

Optimal aggregate testing using Vandermonde polynomials and spectral methods.

Recycled aggregate (RA) has been used in various construction applications around the world mainly as sub-grade, roadwork and unbound materials, but not in higher-grade applications. The major barrier encountered is the variation of quality within RA, which causes lower strength, and poorer quality. This work studies the relationships among six parameters describing the characteristics of RA: (i) particle size distribution, (ii) particle density, (iii) porosity and absorption, (iv) particle shape, (v) strength and toughness, and (vi) chemical composition. Samples of RA from 10 demolition sites were obtained with service life ranging from 10 to 40 years. One additional set of samples was specifically collected from the Tuen Mun Area 38 Recycling Plant. The characteristics of these eleven sets of samples were then compared with normal aggregate samples. A Vandermonde matrix for interpolation polynomial coefficient estimation is used to give detailed mathematical relationships among pairs of samples, which can be used to work out redundant tests. Different orders of interpolation polynomials are used for comparison, hence the best-fit equations with the lowest fitting errors from different orders of polynomials can be found. Fitting error distributions are then studied by using spectral methods such as power spectra and bispectra. From that, the best equations for result estimations can be obtained. This study reveals that there is strong correlation among test parameters, and by measuring two of them: either "particle density" or "porosity and absorption" or "particle shape" or "strength and toughness", and "chemical content", it is sufficient to study RA.

Absorption↗

Flow cytometric, phase-resolved fluorescence measurement of propidium iodide uptake in macrophages containing phagocytized fluorescent microspheres.

BACKGROUND: Spectral interference (overlap) from phagocytosed green-yellow (GY) microspheres in the flow cytometric, red fluorescence emission measurement channel causes errors in quantifying damaged/dead alveolar macrophages by uptake of propidium iodide. METHODS: Particle burdens of uniform GY fluorescent microspheres phagocytosed by rat alveolar macrophages and the discrimination of damaged/dead cells as indexed by propidium iodide uptake were assessed with conventional and phase-sensitive flow cytometry. RESULTS: The fluorescence spectral emission from phagocytosed microspheres partly overlapped the propidium iodide red fluorescence emission and interfered with the measurement of damaged/dead cells when using conventional flow cytometry without subtractive compensation. This caused errors when estimating the percentage of nonviable, propidium iodide-positive, phagocytic macrophages. The interference was eliminated by employing phase-sensitive detection in the red fluorescence measurement channel based on differences in fluorescence lifetimes between the fluorescent microspheres and propidium iodide. Intrinsic cellular autofluorescence, whose fluorescence lifetime is approximately the same as that of the phagocytosed microspheres, also was eliminated in the phase-sensitive detection process. Because there was no detectable spectral interference of propidium iodide in the green fluorescence (phagocytosis) measurement channel, conventional fluorescence detection was employed. CONCLUSIONS: Phase-resolved, red fluorescence emission measurement eliminates spectral overlap errors caused by autofluorescent phagocytes that contain fluorescent microspheres in the analyses of propidium iodide uptake. Cytometry 39:45-55, 2000. Published 2000 Wiley-Liss, Inc.

Animals↗

Extracting information from the power spectrum of synaptic noise.

In cortical neurons, synaptic "noise" is caused by the nearly random release of thousands of synapses. Few methods are presently available to analyze synaptic noise and deduce properties of the underlying synaptic inputs. We focus here on the power spectral density (PSD) of several models of synaptic noise. We examine different classes of analytically solvable kinetic models for synaptic currents, such as the "delta kinetic models," which use Dirac delta functions to represent the activation of the ion channel. We first show that, for this class of kinetic models, one can obtain an analytic expression for the PSD of the total synaptic conductance and derive equivalent stochastic models with only a few variables. This yields a method for constraining models of synaptic currents by analyzing voltage-clamp recordings of synaptic noise. Second, we show that a similar approach can be followed for the PSD of the the membrane potential (Vm) through an effective-leak approximation. Third, we show that this approach is also valid for inputs distributed in dendrites. In this case, the frequency scaling of the Vm PSD is preserved, suggesting that this approach may be applied to intracellular recordings of real neurons. In conclusion, using simple mathematical tools, we show that Vm recordings can be used to constrain kinetic models of synaptic currents, as well as to estimate equivalent stochastic models. This approach, therefore, provides a direct link between intracellular recordings in vivo and the design of models consistent with the dynamics and spectral structure of synaptic noise.

Animals↗

Local image structures and optic flow estimation.

Different kinds of local image structures (such as homogeneous, edge-like and junction-like patches) can be distinguished by the intrinsic dimensionality of the local signals. Intrinsic dimensionality makes use of variance from a point and a line in spectral representation of the signal in order to classify it as homogeneous, edge-like or junction-like. The concept of intrinsic dimensionality has been mostly exercised using discrete formulations; however, recent work has introduced a continuous definition. The current study analyzes the distribution of local patches in natural images according to this continuous understanding of intrinsic dimensionality. This distribution reveals specific patterns than can be also associated to local image structures established in computer vision and which can be related to orientation and optic flow features. In particular, we link quantitative and qualitative properties of optic-flow error estimates to these patterns. In this way, we also introduce a new tool for better analysis of optic flow algorithms.

Algorithms↗

Orienting response and frontal midline theta activity: a somatosensory spectral perturbation study.

OBJECTIVE: Brain electric activity in the theta frequency band has been associated with the encoding of new, and the retrieving of previously stored, information. We studied the time course of stimulus-to-stimulus changes of theta activity under repetitive somatosensory stimulation. MATERIALS AND METHODS: Twelve healthy subjects participated in the study. Repetitive electric stimuli, grouped into 48 stimulus trains, were applied to the left index finger. The stimulus trains contained 27 stimuli (0.9 Hz, 2.5 times sensory threshold). Each stimulus train of 30 s was followed by a stimulus-free break of 30 s. This stimulation paradigm allowed the separate estimation of effects for each position of the stimulus in the train and an analysis of stimulus-to-stimulus changes. Multichannel EEG recordings allowed a topographic analysis of the event-related spectral perturbation effects in the theta frequency band. The brain electric novelty response triggered by the stimulus train onset was analyzed by 3 methods: (1) event-related potentials; (2) event-related power spectra for the investigation of spectral perturbation effects on theta activity; and (3) an approach to break down the stimulus-induced theta activity into phase-locked activity and effects on the spontaneous, ongoing theta activity using digital filtering. RESULTS: The main findings are a frontal midline activation in the theta band with the beginning of the stimulus train, which habituates during the subsequent stimulation cycles, as well as evidence that distinct effects of the first stimulus on the ongoing, non-phase-locked, theta activity exist.

Adult↗

Studies of plasma membrane mechanics and plasma membrane-cytoskeleton interactions using optical tweezers and fluorescence imaging.

We use optical tweezers in conjunction with an optical position-sensing system, which spectrally filters signals generated by a trapped fluorescent microsphere to study plasma membrane (PM) mechanics and its interactions with cytoskeleton. We dynamically measure the PM tethering force on human embryonic kidney cells that are a standard cultured cell line. Recorded tethering force vs. PM displacement profiles, revealed the tether formation process, initiated with linear deformation of the PM, followed by a nonlinear regime and terminated with the local separation of PM. Tethering force vs. displacement profiles were used to estimate tether formation force and stiffness parameter of the PM. Integration of the force-displacement profiles yielded the work of tether formation, including linear and nonlinear components. Our results demonstrate that spectral filtering of the optically trapped fluorescent microsphere image formed on the position-sensing system overcomes the artifacts introduced by the transillumination imaging and allows accurate measures of PM mechanics before and during the initial stages of tether formation.

Biomechanical Phenomena↗

Spectral analysis of heart rate variability as a quantitative measure of parasympatholytic effect--integrated pharmacokinetics and pharmacodynamics of three anticholinergic drugs.

The time course and concentration-effect relationship of parasympatholytic effects of three anticholinergic drugs were investigated using spectral analysis of heart rate (HR) variability. Single intravenous (i.v.) doses of atropine (10 microg/kg), glycopyrrolate (5 microg/kg), scopolamine (5 microg/kg), and placebo were given to eight healthy volunteers in a double-blind, randomized cross-over study. Electrocardiogram (ECG) was recorded at baseline and 2.5, 5, 10, 20, and 30 minutes, and 1, 1.5, 2, 3, 4, 5, and 6 hours after drug administration, while the subjects breathed at a fixed 0.25 Hz frequency. The powers of two frequency bands (low frequency [LF] = 0.07-0.15 Hz and high frequency [HF] = 0.15-0.40 Hz) were calculated using stationary time series of R-R intervals (RRI) free from ectopic beats. To perform pharmacokinetic-pharmacodynamic (PK-PD) modeling, venous plasma drug concentrations were measured. Atropine and glycopyrrolate, and, to a lesser extent, scopolamine induced decreases in HF power and increases in LF/HF ratio of HR variability, indicating parasympatholytic activity and corresponding changes in sympathovagal balance. Maximal average decreases in HF power were 99%, 94%, and 82%, respectively, but in two scopolamine subjects, a parasympathomimetic effect was dominant. Interindividual variability was least for the Hayano index of HF power (square root (RRI HF-power)/RRI*100), and profound and consistent decreases were seen after atropine and glycopyrrolate. Pharmacokinetics were best fitted to a two-compartment open model, and effect compartment link modeling using the Hayano index was performed with the atropine and glycopyrrolate data. The best description of the PK-PD relationship for both drugs was achieved using the sigmoidal Emax model. Mean (+/-SD) EC50, sigmoidicity factor (gamma), and equilibration rate constant (k(e0)) estimates were 1.35 (+/-0.27) ng/mL, 6.07 (+/-1.98) and 11.0 (+/-5.28) l/h for atropine and 1.35 (+/-0.49) ng/mL, 4.34 (+/-1.55) and 2.26 (+/-0.81) l/h for glycopyrrolate. Spectral analysis of HR variability appears to be a powerful tool in monitoring parasympatholytic drug activity. A sigmoidal Emax model with an extremely steep concentration-response relationship was revealed for atropine and glycopyrrolate. The effects of scopolamine were more incongruous.

Adult↗

[Complexes of telomeric oligonucleotide d(TTAGGG)4 with the new recombinant protein PGEk--nucleic acid carrier into proliferating cells].

The complexation of the new protein vector PGEk--a carrier of nucleic acids into proliferating cells with phosphodiester d(TTAGGG)4 (TMO) and phosphorothioate Sd(TTAGGG)4 (TMS) telomerase inhibitor oligonucleotides was studied. PGEk molecule, consisting of 64 amino acids, is comprising the sequence of the human epidermal growth factor EGFh which is hydrophobic cell targeting moiety serving for receptor-mediated endocytosis and an NLS (nuclear localization signal) which is hydrophilic serving as a DNA-binding and selective nuclear import moiety. Experiments were carried out in 0.01 M Na-phosphate buffer contained 0.1 M NaCl, pH 7.8 at 37 degrees C. CD spectral analysis revealed that TMO molecules folded back into intramolecular antiparallel G-quadruplex while TMS molecules were represented as unstructured thread. The number of adsorbed PGEk molecules were estimated using PGEk intrinsic fluorescence decrease and fluorescence polarization increase of PGEk under oligonucleotide titration. Adsorption isotherms were plotted in Scatchard coordinates. We have shown that adsorption of the first two PGEk molecules on TMO and TMS occurs noncooperatively with the high association constants K1(TMO) = (7 +/- 1) x 10(7) M(-1) and K1(TMS) = (3 +/- 0.5) x 10(7) M(-1), respectively. Further adsorption up to 5-6 PGEk molecules on TMO occurrs cooperatively with still high association constant K2(TMO) = (4.0 +/- 1.5) x 10(6) M(-1). TMS oligonucleotide binds the third PGEk molecule rather weakly, K2(TMS) = (8 +/- 2) x 10(5) M(-1). CD spectral analysis revealed that G-quadruplex structure formed by TMO have undergone a partial unfolding by binding of PGEk molecules while single-stranded structure formed by TMS was not affected by binding PGEk. Thus, the tertiary structure of DNA and the number of adsorbed PGEk molecules formed biologically active compounds PGEk: TMO and PGEk: TMS were defined, which are able to penetrate through the membrane of proliferating cells and to suppress their growth.

Active Transport, Cell Nucleus↗

Estimates of L:M cone ratio from ERG flicker photometry and genetics.

Estimates of L:M cone ratio for males with normal color vision were derived using the flicker-photometric electroretinogram (ERG). These were obtained by best fitting ERG spectral sensitivity functions to a weighted sum of long (L)- and middle (M)-wavelength-sensitive cone spectral absorption curves. Using the ERG, measurements can be made with extremely high precision, which leaves variation in the wavelength of maximal sensitivity (lambda(max)) of the cone photopigments as the major remaining source of inaccuracy in determining the ratio of cone contributions. Here that source of inaccuracy was largely eliminated through the use of individualized L-cone spectral absorption curves deduced from L-pigment gene sequences. The method was used on 62 normal males as part of an effort to obtain a true picture of how normal variations in L:M cone ratio are distributed. The percentage of L cones in the average eye was 65%L [where %L = 100 X L / (L+M)]. There were huge individual differences ranging from 28%-93%L, corresponding to more than a 30-fold range in L:M ratio (0.4-13). However, the most extreme values were relatively rare; 80% of the subjects fell within +/-15 %L of the mean, corresponding to a 4-fold range in L:M ratio (1-4). The method remedies major weaknesses inherent in earlier applications of flicker photometry to estimate cone ratio; however, it continues to depend on the assumption that the average L cone produces a response with an identical amplitude to that of the average M cone. A comparison of the ERG results with the distribution of cone ratios estimated from cone pigment messenger RNA in cadaver eyes indicates that the assumption generally holds true. However, there may be a small number of exceptions in which individuals have normally occurring (but relatively rare) amino acid substitutions in one of their pigments that significantly affect the physiology of the cone class containing that pigment, so as to reduce the amplitude of its contribution to the ERG. Consistent with this possibility, extreme cone contribution ratios were found to be associated with atypical L-pigment amino acid combinations.

Adolescent↗

Pressure-induced dissociation of brome mosaic virus.

Brome mosaic virus reversibly dissociates into subunits in the pressure range of 600 x 10(5) to 1600 x 10(5) Pa, as demonstrated by studies of the spectral shift of intrinsic fluorescence, of filtration chromatography and of electron microscopy of samples fixed under pressure. Smaller shell particles (T = 1) were detected as intermediates in the dissociation pathway. Dissociation was facilitated by decreasing the concentration, as expected for a multimolecular reaction. The estimated change in volume upon dissociation into 90 dimer particles was -2960 ml/mol. Large increases in the intrinsic fluorescence intensity and in the binding of bis(8-anilinonaphthalene-1-sulfonate) occurred at pressures higher than 1400 x 10(5) Pa. The pressure-dependence profile of the different spectral properties shifted to lower pressures when 5 mM-MgCl2 was included in the buffer or when the pH was raised from 5.5 to 5.9. When the pressure was progressively increased above 1400 x 10(5) Pa, a value that led to 75% dissociation, the capsid subunits lost the ability to reassociate into regular shells and only amorphous aggregates were formed after decompression, as evidenced by both electron microscopy and gel filtration chromatography. The formation of these random aggregates of brome mosaic virus can be explained by a conformational drift of the separated subunits, similar in nature to that found in simpler oligomeric proteins.

Chromatography, High Pressure Liquid↗

New cholinesterase-inhibiting triterpenoid alkaloids from Buxus hyrcana.

The current phytochemical investigation on Buxus hyrcana Pojark. has resulted in the isolation of the triterpenoid alkaloids 1-10. The structures of five new alkaloids, hyrcanone (1), hyrcanol (2), hyrcatrienine (3), N(b)-dimethylcycloxobuxoviricine (4), and hyrcamine (5), were elucidated by means of modern spectroscopic techniques, while the known alkaloids, buxidin (6), buxandrine (7), buxabenzacinine (8), buxippine-K (9) and E-buxenone (10), were identified by comparing their spectral data with those reported earlier. Compounds 1 and 3-9 were found to be acetyl- and butyrylcholinesterase inhibitors. The IC50 values were estimated to be in the range of 83.0-468.0 microM against AChE and 1.12-350.0 microM against BChE. The structure-activity relationship studies suggested that the presence of dimethylamino moieties at C(3) and C(20) is the most important factor influencing the activity of these compounds against the cholinesterase enzymes. All compounds were also evaluated for cytotoxicity on a fibroblast cell line with incubation of 24 h. No cytotoxic effects were exerted by any compound.

Acetylcholinesterase↗

Functional high level expression of cytochrome P450 CYP2D6 using baculoviral expression systems.

Cytochrome P-450 CYP2D6 plays a central role in the metabolism of many widely used therapeutic drugs including beta-adrenergic antagonists, antiarrhythmics, and tricyclic antidepressants. Recombinant baculoviruses have been constructed containing the full-length human CYP2D6 cDNA and used to express CYP2D6 in Spodoptera frugiperda (Sf9) cells. High levels of recombinant protein have been produced using either polyhedrin or basic protein promoters (0.05-0.20 nmol/mg cell protein; 0.05-0.15 nmol/liter). The enzyme is catalytically active toward CYP2D6 substrates such as bufuralol and metoprolol. In order to optimize catalytic activity human reductase was coexpressed with CYP2D6 in Sf9 cells; reductase activity was in the region of 1000-1500 units per mg cell protein, while spectrally active CY2D6 was in the range 10-20 pmol/mg cell protein. The K(m) and K(cat) values for bufuralol metabolism were estimated as 4.7 microM and 12.23 min-1, respectively. The use of the conventional very late promoters such as the polyhedrin promoter generate a large proportion of inactive CYP2D6. The problem was to a degree circumvented using the "late" basic promoter which is active earlier in the baculovirus infection cycle. The yield of functional CYP2D6 was at least as high as with very late promoters, but the proportion of inactive protein was reduced. Bufuralol hydroxylase activity could be measured directly by HPLC analysis of cell culture media supplemented with bufuralol, and we have developed a plate assay system which provides a simple method for the analysis of drug metabolism reactions using Sf9 cells. Expression using baculovirus provides a valuable source of functional CYP2D6 for work aimed at elucidating the structure and function of the enzyme.

Adrenergic beta-Antagonists↗

31P NMR and freeze fracture studies of sarcoplasmic reticulum membranes from normal and malignant hyperthermic pigs: effect of halothane and dantrolene.

The effects of halothane and dantrolene on sarcoplasmic reticulum membranes isolated from normal and malignant hyperthermia pig muscle have been investigated using 31P NMR and freeze fracture electron microscopy. The dynamical and structural changes are estimated from the second moment, as calculated from 31P NMR spectra. For both membranes, addition of halothane induces a similar decrease in the spectral second moment. At high concentration of halothane, freeze fracture replicas show small unilamellar vesicles or mixed micelles, uniformly sprayed in the case of malignant hyperthermia membranes but mainly aggregated for the normal ones. The effect of halothane on both membranes is partially inhibited by adding dantrolene. These results suggest that (i) the malignant hyperthermia syndrome is not directly related to the polar heads of phospholipids and (ii) dantrolene counteracts unspecifically the disturbing effect of halothane at the lipid level.

Animals↗

The effect of experimental muscle pain on the background electrical brain activity.

The purpose of this project was to investigate whether specific effects in the background activity of the brain associated with the experience of pain can be depicted by means of quantitative electroencephalography (EEG). Lasting pain was induced by intramuscular infusion of hypertonic saline. The infusion was titrated to maintain pain for a sufficient time to obtain enough data for meaningful analysis. In a first study on 12 subjects, using a single, blind, repeated measures design with randomization of the administration of isotonic (0.9%) and hypertonic (5%) saline, and with subjects unaware of the fact that one substance was isotonic saline, a statistically significant pain response could be attributed to the administration of hypertonic saline. In a second study on 19 subjects, again using a randomized repeated measures design, topographic EEG measures were examined with respect to experimentally induced pain and pain from memory. Prior to each of these experimental stages, baseline recordings were obtained to satisfy the requirement of the crossover design. In addition to the common frequency bands used in EEG, we also obtained data in the frequency range of 35-100 Hz. The short-term variability of the selected EEG measures and their suitability as a sample estimate were assessed by computing the coefficient of variation from all selected epochs of a given subject at baseline. When compared to baseline, spectral analyzed EEG measures during experimental pain demonstrated statistically significant increases in the beta and 35-100 Hz frequency ranges, most notably at the temporal recording sites. There was no statistically significant difference between the EEG measures for (1) experimental pain vs. pain from memory, and (2) the 2 baseline recordings. The great variability in the topographical aspect of the between-subject response was interpreted as being strongly suggestive of the contamination of EEG measures by phenomena attributed to the jaw, facial and scalp musculature. In fact, Pearson correlation coefficients, as high as 0.92, were found between measures in the frequency band of 35-100 Hz and the beta frequency range. The unexplained variance in the heightened beta cortical power density can be attributed to the vigilance scanning of pain processes. Due to the fact that the statistically significant effect of pain on the topographic EEG measures were not different from imagined pain, we concluded that these effects are non-specific for pain.

Adult↗

Frequency selectivity in workers with noise-induced hearing loss.

This study was undertaken in order to document, in a group of subjects affected by a noise-induced hearing loss, the relation between the loss of auditory sensitivity and the loss of frequency selectivity at mid-frequencies, namely 1000 and 3000 Hz. Auditory filter shapes were estimated using the notched noise method. Twelve notch widths were tested, six symmetrical and six asymmetrical with respect to the signal frequency; the spectral level of the noise was set at 50 dB SPL. Data were collected with 22 noise-exposed workers having different degrees of hearing loss. The findings indicate that above a certain degree of hearing loss, which seems to be around 30 dB HL, frequency selectivity tends to decrease linearly with increase in loss of sensitivity. Even when the degree of hearing loss is similar in origin and in magnitude, there is a wide variation among subjects in auditory filter bandwidth. Based on the data collected in this study, it is not possible to adequately predict the auditory filter bandwidth of an individual from hearing threshold levels.

Acoustic Stimulation↗

Learning subject-specific spatial and temporal filters for single-trial EEG classification.

There are a wide variety of electroencephalography (EEG) analysis methods. Most of them are based on averaging over multiple trials in order to increase signal-to-noise ratio. The method introduced in this article is a single trial method. Our approach is based on the assumption that the "response of interest" to each task is smooth, and is contained in several sensor channels. We propose a two-stage preprocessing method. In the first stage, we apply spatial filtering by taking weighted linear combinations of the sensor measurements. In the second stage, we perform time-domain filtering. In both steps, we derive filters that maximize a class dissimilarity measure subject to regularizing constraints on the total variation of the average estimated signal (or, alternatively, on the signal's strength in time intervals where it is known to be absent). No other spatial or spectral assumptions with regard to the anatomy or sources were made.

Algorithms↗

Generation and characterization of correlated spike trains.

A framework within which to generate and characterize populations of correlated spike trains is presented. Spike trains are generated using integrate to threshold and fire type encoders. Spectral analysis techniques form the basis for specifying the strength of correlation within the population. To accurately specify weak correlation, a combined coherence estimate is formed between several independent pairs of spike trains from a population. The aim is to produce populations of spike trains which have a realistic stochastic correlation structure and which can be used to explore the role of correlated neuronal discharge in information processing in neuronal systems.

Computer Simulation↗