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Effect of oxidative sulfitolysis of disulfide bonds of bovine serum albumin on its structural properties: a physiochemical study.

The effect of S-S bond cleavage of bovine serum albumin (BSA) on some of its structural properties, including solubility, viscosity, and conformation, were investigated. Cleavage of S-S bonds decreased the solubility of serum albumin and also shifted its isoelectric point to lower pH values. S-S bond cleavage resulted in changes in shape and hydrodynamic volume of the protein, increasing the specific viscosity, with cleavage of up to 14 S-S bonds, followed by a decrease with further cleavage. Both UV difference and fluorescence spectral measurements indicated that conformational flexibility increases with S-S bond cleavage. Secondary structure estimations by far UV-CD suggested a gradual decrease in alpha-helical content of the protein with progressive cleavage of its S-S bonds. However, fully S-S bond cleaved protein maintained some alpha-helical structure. Sulfitolysis of the protein also decreased its 1,8-anilino-naphthalene sulfonate-binding ability.

Circular Dichroism↗

Dynamic power and coherence analysis of ultra short-term cognitive processes--a methodical study.

This EEG mapping study was designed to explore ultra short-term cognitive processes in human thinking. The EEG was recorded while the subjects performed two tasks randomly mixed in time: category concept activation and pattern comparison. A novel approach is introduced to examine these tasks by means of band powers and coherence with high time and frequency resolution. This approach is based on general adaptive principles and the adaptive fit of a bivariate linear model (ARMA model) with time varying parameters, and allows the estimation of band powers and coherences continuously in time. The application of mapping to these spectral parameter functions results in map sequences of band powers, and local band coherences. It was proved that these map sequences of the frequency band 13-20Hz reflect the dynamic behaviour of information processing in a characteristic topographical manner. Based on these dynamic topographical examinations, conceptual and imaginal representations are distinguishable.

Brain Mapping↗

A comparison of analytical methods for the study of fractional Brownian motion.

Fractional Brownian motion (FBM) provides a useful model for many physical phenomena demonstrating long-term dependencies and l/f-type spectral behavior. In this model, only one parameter is necessary to describe the complexity of the data, H, the Hurst exponent. FBM is a nonstationary random function not well suited to traditional power spectral analysis however. In this paper we discuss alternative methods for the analysis of FBM, in the context of real-time biomedical signal processing. Regression-based methods utilizing the power spectral density (PSD), the discrete wavelet transform (DWT), and dispersive analysis (DA) are compared for estimation accuracy and precision on synthesized FBM datasets. The performance of a maximum likelihood estimator for H, theoretically the best possible estimator, are presented for reference. Of the regression-based methods, it is found that the estimates provided by the DWT method have better accuracy and precision for H > 0.5, but become biased for low values of H. The DA method is most accurate for H < 0.5 for a 256-point data window size. The PSD method was biased for both H < 0.5 and H > 0.5.

Bias↗

Accumulation of protoporphyrin-IX by the chlorophyll-less y-y mutant of Chlamydomonas reinhardtii.

A pigment accumulating in a Mendelian mutant (y-y) of Chlamydomonas reinhardtii, which has essentially no chlorophyll and lacks inner chloroplast membranes in the light and dark, was isolated and characterized. It was identified as protoporphyrin-IX (PROTO) by spectral analysis using two different methods of extraction and fractionation. The amount of PROTO was estimated to be 10(7) molecules per cell. Since PROTO was the only intermediate of chlorophyll biosynthesis that accumulated, we conclude the y-y lesion in the pathway is after PROTO.

Chlamydomonas↗

In vivo demonstration of flow recirculation and turbulence downstream of graded stenoses in canine arteries.

The velocity field around arterial stenoses was investigated using a pulsed doppler velocimeter in vivo. Asymmetric zones of recirculation were identified by systolic flow reversal in the post-stenotic field in carotid and iliac arteries of anesthetised dogs. There was a close correlation between shear intensity and turbulence as estimated by the velocity difference between the jet and the recirculation zone and by maximum spectral width respectively. Under the conditions of these experiments, stenosis grade (% diameter reduction) dominated hemodynamic variables such as Reynolds number, oscillation and pulsatility in determining the intensity of turbulence. The method used does not appear to have sufficient resolution to distinguish between turbulence and discrete oscillating velocities (vorticity) nor to allow determination of wall shear stress though the pattern of change of the latter is similar to that found downstream of axisymmetric stenosis in models using steady flow.

Animals↗

Assessment of composition and anisotropic elastic properties of secondary osteon lamellae.

Measurement of the elastic properties of single osteon lamellae is still one of the most demanding tasks in bone mechanics to be solved. By means of site-matched Raman microspectroscopy, acoustic microscopy and nanoindentation the structure, chemical composition and anisotropic elasticity of individual lamellae in secondary osteons were investigated. Acoustic impedance images (911-MHz) and two-dimensional Raman spectra were acquired in sections of human femoral bone. The samples were prepared with orientations at various observation angles theta relative to the femoral long axis. Nanoindentations provided local estimations of the elastic modulus and landmarks necessary for spatial fusion of the acoustic and spectral Raman images. Phosphate nu(1) (961 cm(-1)) and amide I (1665 cm(-1)) band images representing spatial distributions of mineral and collagen were fused with the acoustic images. Acoustic impedance was correlated with the indentation elastic modulus E(IT) (R(2)=0.61). Both parameters are sensitive to elastic tissue anisotropy. The lowest values were obtained in the direction perpendicular to the femoral long axis. Acoustic images exhibit a characteristic bimodal lamellar pattern of alternating high and low impedance values. Since this undulation was not associated with a variation of the phosphate nu(1)-band intensity in the Raman images, it was attributed to variations of the lamellar orientation. After threshold segmentation and conversion to elastic modulus the orientation and transverse isotropic elastic constants were derived for individual ensembles of apparent thin and thick lamellae. Our results suggest that this model represents the effective anisotropic properties of an asymmetric twisted plywood structure made of transverse isotropic fibrils. This is the first report that proves experimentally the ability of acoustic microscopy to map tissue elasticity in two dimensions with micrometer resolution. The combination with Raman microspectroscopy provides a unique way to study bone and mineral metabolism and the relation with mechanical function at the ultrastructural tissue level.

Anisotropy↗

Low temperature FTIR spectra and hydrogen bonds in polycrystalline cytidine.

FTIR spectra of polycrystalline samples of cytidine, pure and containing a small quantity of N(O)H or N(O)D groups (<20%), were measured in KBr pellets from 4000 to 400 cm(-1) at temperatures from 300 to 20K. For the first time the bands of the narrow isotopically decoupled proton stretching vibration mode (nu(1)) of OH- and NH- groups were found; their number corresponds to the number of H-bonds in crystal according to structural data. The FTIR spectra at low temperature in the out-of-plane bending nu(4) proton mode range (lower than 1000 cm(-1)) of N(O)H groups revealed narrow bands, which correspond to nu(1) bands together with several "extra" bands, which are influenced by the isotopic exchange and (or) cooling. All of them have their counterparts in the N(O)D-substance spectrum with an isotopic frequency ratio of 1.30-1.40. The "extra" bands are assigned to the H-bound OH and NH protons, which are disordered and cannot be seen with X-ray crystal structure analysis. The peak positions of both mode bands (expressed as the red shift of nu(1) or blue shift of nu(4) modes relatively free molecules) were used for the estimation of the energy of different H-bonds using previously established empirical correlations between spectral and thermodynamic parameters of hydrogen bonds. The correlation of the red shift and H-bond length is also confirmed for all five H-bonds of cytidine.

Cold Temperature↗

A comparison of spectral edge, delta power, and bispectral index as EEG measures of alfentanil, propofol, and midazolam drug effect.

BACKGROUND: The effects of anesthetic drugs on electroencephalograms (EEG) have been studied to develop the EEG as a measure of anesthetic depth. Bispectral analysis is a new quantitative technique that measures the consistency of the phase and power relationships and returns a single measure, the bispectral index. The purpose of this study was to compare the performance of the bispectral index, version 1.1, with other spectral analysis EEG measures of drug effect for three commonly used anesthetic drugs. METHODS: The EEG waveforms from 31 adults receiving infusions of alfentanil, propofol, or midazolam were analyzed. The time course of spectral edge (SE95), relative power in delta band, and bispectral index were related to the estimated effect-site concentration with use of a sigmoidal Emax model to estimate the potency (IC50) and the plasma effect-site equilibration rate constant (Ke0) for each measure. The performance of the fitting was assessed by the coefficient of correlation between predicted and observed effect. RESULTS: Alfentanil induced a high-amplitude low-frequency EEG response. Propofol induced a biphasic response. At low concentrations, both frequency and amplitude increased. When the concentration increased, the EEG slowed and the amplitude decreased. High concentration produced burst suppression. Midazolam increased EEG frequency and amplitude. Bispectral index, SE95, and delta power yield similar estimates of IC50 and ke0. Except for alfentanil, the performance of the modeling with the bispectral index was as good that with SE95 or delta power. CONCLUSION: Bispectral analysis can be used as a measure of the EEG effects of anesthetic drugs.

Adult↗

Computer-assisted spectral electroretinography in vitrectomy patients.

Computer-assisted spectral electroretinography (CASE), using full field stimulation in the presence of a rod saturating background light, has been found to be a useful means of evaluating retinal function in patients with vitreal opacities. By this means, cone electroretinograms (ERGs) to white as well as spectrally selective stimuli could easily be detected in 22 of 24 eyes with vitreal opacities obscuring any view of the fundus and with minimal or no detectable ERGs to standard single flash stimuli. The responses to spectral stimuli in the presence of a rod saturating background light provides a means of estimating the optical density of the opacity. The results indicate that most patients undergoing vitrectomy for such opacities have considerable superimposed retinal damage and that the opacities themselves are seldom sufficiently dense to decrease the ERG to any great extent. Five eyes studied before and after vitrectomy indicate that successful vitrectomy has no effect on the ERG. The results suggest that CASE may be a useful substitute to the bright flash ERG for evaluating global retinal function behind an opaque media.

Adolescent↗

The analysis of animal bioelectric brain activity influenced by microwaves or by the introduction of strychnine.

The widespread impact made by technology has raised concerns about the safety of human exposure to electromagnetic radiation in the environment. The brain is especially sensitive to the influence of microwaves. The most effective method for estimation of the organism's functional states is an analysis of electroencephalograms. The statistical and spectral methods are usually used for analysis of animal electrocorticograms. The information obtained in such way is the integrated character and it is sometimes insufficient for identification of the brain state charging caused by various factors, especially microwaves altering the ecological situation. The nonlinear dynamics method is used in our work concurrent with the spectral correlation method for animal electrocorticogram processing. The correlation dimensionality represents a numerical criterion allowing for comparative investigation of various dynamic states of the system. In the process of investigation, it has been found that the nonlinear dynamics method may be used to analyze the electrocorticograms of experimental animal in different functional states being confirmed by increasing parameter of the correlation dimensionality in electrocorticograms of animal irradiated by microwaves or subjected to the introduction of strychnine.

Algorithms↗

Reduced vagal activity in salt-sensitive subjects during mental challenge.

BACKGROUND: Salt-sensitive normotensive men exhibit an enhanced pressor and heart rate (HR) response to mental stress. Stress-induced HR acceleration may result from sympathetic activation or vagal withdrawal. We studied the importance of vagal withdrawal for the increased stress responsiveness of salt-sensitive subjects. METHODS: We studied cardiovascular reactivity to mental challenge in 17 salt-sensitive healthy white male students and 56 salt-resistant control subjects who were comparable with respect to age, body mass index, and physical fitness. Salt sensitivity was determined by a 2-week dietary protocol (20 mmol v 240 mmol sodium/day). Mental stress was induced by a computerized information-processing task (manometer test). Electrocardiogram and finger blood pressure (BP; Finapres, Ohmeda, Louisville, CO) were registered continuously to determine HR and interbeat-interval length. Time and frequency domain (spectral power) based measures of respiratory-related heart rate variability (HRV) were calculated to estimate vagal cardiac control; diastolic BP reactivity was assessed to estimate peripheral sympathetic effects. RESULTS: Stress-induced increase in HR was higher in salt-sensitive than in salt-resistant subjects. Salt-sensitive subjects, in comparison to salt-resistant subjects, showed significantly reduced respiratory-related HRV during baseline and mental stress conditions (P <.01). The increase in diastolic BP during mental challenge was significantly greater in salt-sensitive subjects (P <.05). CONCLUSIONS: Our findings suggest reduced vagal and increased sympathetic tone during mental challenge in salt-sensitive subjects. Altered autonomic nervous system function may contribute to later development of hypertension in salt-sensitive individuals.

Adult↗

Fiber-optic evanescent-field laser sensor for in-situ gas diagnostics.

A compact, rugged and portable fiber-optic evanescent-field laser sensor is developed for the detection of gaseous species in harsh environments such as volcano fumaroles or industrial combustion of glass furnaces. The sensor consists of an optical multi-mode fused silica fiber with jacket and cladding removed and the bare fiber core in direct contact with the surrounding molecules. The beam of a single-mode DFB diode laser with an emission wavelength centered at 1.5705 microm is coupled into the fiber. At the other end of the fiber an infrared detector is used to record the transmitted infrared laser light intensity. Due to the frustrated total reflection (FTR) and the attenuated total reflection (ATR) the laser intensity is attenuated when passing through the fiber. The FTR is related to a change of the index of refraction while the latter one is related to a change of the absorption coefficient. While tuning the DFB laser wavelength across absorption lines of molecules surrounding the fiber a spectral intensity profile is measured. Voigt functions are fitted to the recorded intensity profiles to estimate relative molecule concentrations. In this paper results from first field measurements at the volcano site 'Solfatara' in Italy are reported that use such a sensor device for simultaneous detection of H2S, CO2 and H2O directly in the gas stream of a volcano fumarole.

Carbon Dioxide↗

Phase relationships between cortical and muscle oscillations in cortical myoclonus: electrocorticographic assessment in a single case.

AIM: To compare voluntary- and sensory-induced myoclonic jerks using spectral analysis in a subject with cortical myoclonus. METHODS: The coherence, phase and cumulant density estimates were calculated between right electrocorticographic (ECoG) signals and distal left leg muscles in a patient with subdural electrodes inserted over the right sensorimotor cortex. RESULTS: Significant coherence between sensorimotor cortex and muscle was found up to 60 Hz during voluntary induced myoclonic jerks. Additional higher frequency coherence ( approximately 140 and 190 Hz) was found during sensory-induced myoclonic jerks. The cortical signals phase led muscle signals at frequencies >15 Hz by delays consistent with transmission along corticospinal pathways. Below 15 Hz the cortex phase lagged the muscle signals. Polarity reversal of the cumulant density estimate and the ECoG site demonstrating the highest coherence helped to localize the site of the abnormal oscillatory activity to the leg area of the motor cortex. CONCLUSIONS: Oscillations of different frequencies can co-exist at a given location and can both phase lead and lag contralateral muscle. This has implications for cortex-muscle latency measures calculated by back-averaging techniques.

Aged↗

DNA sequencing by capillary electrophoresis: use of a two-laser-two-window intensified diode array detection system.

This paper presents the principles of an instrument designed for DNA sequencing using the standard four-dye-labeled primer approach. The method is based on capillary electrophoresis with laser-induced fluorescence and an intensified diode array detector. An important goal of the instrument design has been a detection system that possesses high sensitivity and high spectral resolution. Based on an analysis of the spectral characteristics of the four standard dye-labeled primers, FAM, JOE, ROX, and TAMRA, the strategy has been to use a two-laser-two-window approach, in which a 488-nm argon ion laser illuminates one window, followed by a 543-nm helium-neon laser illuminating the second window. The two-window approach has no moving parts and permits continuous illumination. Spectral resolution is provided by a grating spectrograph and a cooled intensified diode array. The estimated limit of detection for the standard four dye-labeled primers was found to be in the sample concentration range of 1 x 10(-12) M. To achieve these low levels, complete free-radical polymerization of polyacrylamide has been found to be necessary in order to reduce background noise. In addition, reduction in background noise was accomplished by continual purging of the anodic reservoir in order to prevent electrolysis products from entering the capillary. Separation of DNA sequencing reaction products is demonstrated on a 9% T linear polyacrylamide column.

Base Sequence↗

Distances among coenzyme and metal sites of NADP+-dependent isocitrate dehydrogenase using resonance energy transfer.

When the substrate isocitrate-Mn2+ is present, the fluorescent nucleotide analogue 2-[(4-bromo-2,3-dioxobutyl)thio]-1,N6-ethenoadenosine 2',5'-bisphosphate (2-BDB-T epsilon A-2',5'-DP) reacts irreversibly with pig heart NADP+-specific isocitrate dehydrogenase at the coenzyme binding site on one subunit of the dimeric enzyme [Bailey, J. M., & Colman, R. F. (1985) Biochemistry 24, 5367-5377]. The modified enzyme, which retains partial activity, binds 1 mol of NADPH or 1 mol of the coenzyme analogue, reduced thionicotinamide adenine dinucleotide phosphate (TNADPH), per dimer. TNADPH quenches the fluorescence of enzyme-bound 2-BDB-T epsilon A-2',5'-DP with an efficiency of energy transfer of 9.8%. From this value and the spectral properties of the donor and acceptor chromophores, a distance of 32 A was calculated as the average distance between coenzyme sites on the two subunits. Isocitrate dehydrogenase activity requires a divalent metal ion, such as Mn2+, Co2+, or Ni2+. Co2+ and Ni2+ have absorption spectra that overlap the emission spectra of enzyme-bound 2-BDB-T epsilon A-2',5'-DP. In the presence of isocitrate, each of these two metal ions quenches the fluorescence of the enzyme-bound reagent with an efficiency of energy transfer of 28-29%. From this value and the spectral characteristics of the energy donor and acceptors, an average distance of 8.0 A was estimated between the metal-isocitrate site and the labeled coenzyme site. These distances have provided constraints in formulating a model of the spatial arrangement of active-site ligands on isocitrate dehydrogenase.

Adenosine Diphosphate↗

Influence of the preparation method on the physicochemical properties of tolbutamide/cyclodextrin binary systems.

Tolbutamide (TBM) was found to form an inclusion complex with beta cyclodextrin (beta-CD) in solution and in solid state. Inclusion complex formation between tolbutamide and beta-cyclodextrin in aqueous solution was studied by phase solubility and spectral shift methods. The apparent stability constant Ks calculated by these techniques, in water, were estimated as 195.7 and 236.5 M(-1), respectively. The phase solubility studies revealed a B(S)-type diagram with an inclusion complex of 1:2 molar ratio. The solid inclusion complexes of TBM and beta-CD were prepared at a molar ratio of 1:2 by kneading, coprecipitation, freeze-drying, and spray-drying methods. In addition, the physical mixture was prepared. Characterization of TBM: beta-CD inclusion was performed using differential scanning calorimetry (DSC), Raman spectroscopy, and X-ray diffractometry and by application of a so-called ether wash method. All the inclusion systems investigated led to a significant improvement in the dissolution over free TBM, and the dissolution rate of the active material was observed to be independent of the preparation method.

Calorimetry, Differential Scanning↗

Bulk optical properties of healthy female breast tissue.

We have measured the bulk optical properties of healthy female breast tissues in vivo in the parallel plate, transmission geometry. Fifty-two volunteers were measured. Blood volume and blood oxygen saturation were derived from the optical property data using a novel method that employed a priori spectral information to overcome limitations associated with simple homogeneous tissue models. The measurements provide an estimate of the variation of normal breast tissue optical properties in a fairly large population. The mean blood volume was 34 +/- 9 microM and the mean blood oxygen saturation was 68 +/- 8%. We also investigated the correlation of these optical properties with demographic factors such as body mass index (BMI) and age. We observed a weak correlation of blood volume and reduced scattering coefficient with BMI: correlation with age, however, was not evident within the statistical error of these experiments. The new information on healthy breast tissue provides insight about the potential contrasts available for diffuse optical tomography of breast tumours.

Adult↗

Neurotoxic esterase (NTE) assay: optimized conditions based on detergent-induced shifts in the phenol/4-aminoantipyrine chromophore spectrum.

An assay for neurotoxic esterase (neuropathy target esterase, NTE) was developed by Johnson (1,2) to assess the delayed neurotoxic potential of organophosphorus compounds. NTE activity is calculated from the rate of phenyl valerate hydrolysis resistant to paraoxon and sensitive to mipafox inhibition under specified conditions of inhibitor concentrations, pH, temperature, and incubation times with inhibitors and substrate. The amount of phenol produced is measured colorimetrically after its oxidative coupling with 4-aminoantipyrine to yield 4-N-(1,4-benzoquinoneimine)-antipyrine, a chromophore with a wavelength of maximum absorbance (lambda m) 510 nm and corresponding molar absorptivity (molar extinction coefficient, epsilon) equal to 13,900 M-1cm-1. The assay was improved and simplified later by Johnson (3) without any change in the lambda m or epsilon, even though the chromophore solvent was altered by adding the detergent, sodium dodecyl sulfate (SDS). The present work demonstrates that when the NTE assay is performed according to the improved procedure, with a final [SDS] of 3.0 mg/mL, the lambda m of the chromophore in the assay mixture is shifted from 510 to 490 nm. The same shift in the chromophore lambda m is observed when phenol standards are coupled with 4-aminoantipyrine in solutions containing 3.0 mg/mL SDS. A systematic investigation of the dependence of the lambda m of the chromophore on [SDS] in the assay mixture revealed that the spectral shift increases rapidly at an [SDS] greater than the apparent critical micelle concentration (CMC; estimated to be 0.53 mg/mL under these conditions) and begins to plateau at [SDS] greater than 10 mg/mL.(ABSTRACT TRUNCATED AT 250 WORDS)

Ampyrone↗