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The conformations of a functional spin-labeled derivative of gastric H/K-ATPase investigated by EPR spectroscopy.

A spin-labeled derivative of porcine gastric H/K-ATPase with high ATP hydrolyzing activity (77 mumol of Pi/(mg.h)) has been prepared. Over 65% of initial ATPase activity (115 mumol of Pi/(mg.h)) was preserved after complete reaction of the enzyme with the lysine reactive nitroxide spin-labeled TEMPO isothiocyanate (TITC). In contrast, rapid and complete loss of ATPase activity occurred after reaction of the enzyme with the lysine directed fluorescent probe FITC. Conventional EPR spectra of TITC labeled H/K-ATPase reflected mainly the slow rotational diffusion of the enzyme in the membrane. An upper limit enzyme intramembranous radius of 108 A was calculated on the basis of rotational correlation times estimated from saturation transfer (ST) EPR spectral lineshapes. Conventional EPR spectra exhibited two major components corresponding to at least two populations of strongly constrained spin-labels. Difference spectroscopy revealed that the proportion of these two components changed markedly with temperature. Moreover, the proportion of the components was sensitive to the presence of the activating ionic ligands Mg2+ and ATP, which induce enzyme conformational transitions, and to the reversible inhibitor SCH 28080, which binds to the K+ sensitive form of the enzyme. These findings show that EPR spectroscopy is able to report functionally coupled conformational changes of gastric H/K-ATPase and imply that the spin-labels are attached to lysines within functionally important regions of the enzyme.

Animals↗

Fluorescence of raw cane sugars evaluated by chemometrics.

In a fluorescence study of raw cane sugar samples, two-way and three-way chemometric methods have been used to extract information about the individual fluorophores in the sugar from fluorescence excitation-emission landscapes. A sample set of 47 raw sugar samples representing a varied selection was analyzed, and three individual fluorophores with (275, 350) nm, (340, 420) nm, and (390, 460) nm as their approximate excitation and emission maxima were found. The spectral profiles of the fluorophores were estimated with the three-way decomposition model PARAFAC. Two-way principal component analysis (PCA) of unfolded fluorescence landscapes confirmed the PARAFAC results and showed patterns of samples related to time of storage. Partial least squares (PLS) calibration models of color at 420 nm had a high model error due to the very high color range of the raw sugars, but variable selection performed on the fluorescence data revealed that all three fluorophores were correlated to color. The (275, 350) nm fluorophore is considered as a color precursor to the color developed on storage and the (340, 420) nm and (390, 460) nm fluorophores show colorant polymer characteristics.

Carbohydrates↗

How well does color perimetry isolate responses from individual cone mechanisms?

PURPOSE: Over the past two decades there has been renewed interest in the use of color perimetry as a means of detecting early functional defects resulting from glaucomatous optic neuropathy and other forms of ophthalmic and neurologic pathology. The most popular form of color perimetry employs a colored background that selectively desensitizes two of the cone mechanisms, while the wavelength of the test target is selected to favor detection by the remaining, relatively unadapted, cone mechanism. While there are data to support the assertion that blue on yellow perimetry adequately isolates the short wavelength sensitive (SWS) cone mechanism, the only estimates of isolation of the other two cone mechanisms are for the region of the visual field corresponding to the fovea. The purpose of this experiment was thus to determine the amount of cone mechanism isolation that is afforded by automated perimetry when using yellow, blue, and magenta adapting backgrounds. METHODS: To estimate cone mechanism isolation, we determined spectral sensitivity for a range of narrow-band stimuli using a standard 30-2 program on a modified Humphrey perimeter. Targets were presented against three different backgrounds of different luminance; yellow at 2.1 log cd.m-2, magenta at 1.3 log cd.m-2, and blue at 1 log cd.m-2. Sensitivity values for each background at each stimulus position were plotted as a function of wavelength, normalized, and then fitted with cone sensitivity templates to determine the relative sensitivity of the three cone mechanisms. RESULTS: The maximum relative isolation of an individual cone mechanism was achieved with a yellow background, where there was an average relative isolation of 0.94 for the SWS cone mechanism; the blue background provided a relative isolation of 0.89 for the long wavelength sensitive (LWS) cone mechanism. The magenta background proved poor at isolating the medium wavelength sensitive (MWS) cone mechanism, where a relative isolation of only 0.51 was obtained. CONCLUSION: Although color perimetry is capable of isolating individual cone mechanisms, the magnitude of isolation in normal observers may be small under certain circumstances. Therefore, when the technique is used to examine pathologic states, it may be necessary to employ at least two target wavelengths to determine the cone mechanism that is performing target detection. Furthermore, we suggest that MWS cone mechanism isolation may be improved through the combination of the so-called silent substitution technique with that of selective adaptation.

Adult↗

The ultimate wavelength for protein crystallography?

This paper describes an analysis of the optimum choice of the X-ray wavelength for macromolecular diffraction data collection. It is shown that there is no ultimate X-ray wavelength for protein crystallography and that the optimum wavelength depends to a large extent on the size of the protein crystal. It also depends on instrumental factors, such as efficiency of the detector for a particular wavelength and spectral density of the synchrotron radiation. Estimates of the optimum wavelength as a function of crystal size are given.

Journal Article↗

Pronounced resting bradycardia in male elite runners is associated with high heart rate variability.

Forty-eight hour Holter monitoring was undertaken of 16 male elite middle- and long-distance runners, age 25 +/- 3 years, with peak oxygen uptake 4.83 +/- 0.43 1 O2/min or 73.0 +/- 3.9 ml O2/kg/min. The athletes had pronounced bradycardia during the night-time, with heart rate calculated from four RR intervals < 30 beats/min in five runners. Twelve of 16 runners had RR intervals > 2 s. Of those, 10 runners had sinus pauses exceeding 2 s, the longest being 3.06 s. Three runners had AV block II, two with Mobitz type 1, and one with both Mobitz type 1 and 2. Autonomic function was estimated by time domain and power spectral analysis of heart rate variability. The runners were compared with a control group of 13 sedentary or moderately active subjects. The runners had a mean of 14 b.p.m. lower heart rate at night than the controls. The runners had higher heart rate variability in all spectral bands. In the time domain pNN50 and rMSSD, which are considered to reflect strongly vagal tone, were markedly higher in the runners than the controls. The findings suggest that an increased parasympathetic tone might at least partly explain the pronounced resting sinus bradycardias found in endurance-trained runners.

Bradycardia↗

Investigation of the effects of temporal and spatial interactions on speech-recognition skills in cochlear-implant subjects.

Forward masking was investigated as a measure of spectral and temporal interactions. Such interactions may adversely affect speech recognition in cochlear-implant subjects. Seven subjects, implanted with the Nucleus 22 device, performed a forward-masking task. They also performed an electrode-discrimination task in order to measure spectral interactions without temporal interactions. Correlation analysis indicated a significant relationship between data obtained in the two tasks (p < 0.1). The two tasks were also correlated with the subjects' scores from five measures of speech recognition. Forward masking and electrode discrimination were strongly correlated with measures requiring consonant and phoneme recognition, respectively. These results indicate that the relationship between forward masking and speech recognition may be due, in part, to a lack of spectral resolution. The data also indicate that consonants may be more readily masked than vowels. Forward-masking data measured for all clinically programmed electrodes in three of the seven subjects were used with a model of the spectral maxima sound processor (SMSP) to estimate the number of electrodes stimulated during a consonant that might be masked by prior presentation of a vowel. These results suggest that temporal interactions across electrodes may be a factor in speech-recognition abilities of some cochlear-implant subjects.

Adult↗

Quantitative ultrasonography in rheumatoid arthritis: evaluation of inflammation by Doppler technique.

OBJECTIVE: To evaluate ultrasonographic methods, including the Doppler technique, as measures of synovial inflammation in finger joints of patients with rheumatoid arthritis. METHODS: Ultrasonography was performed with a high frequency transducer (13 MHz). Evaluation of the sonographic data was conducted by two independent observers and included measurement of synovial area and thickness (grey tone ultrasound), vascularisation (power/colour Doppler), and indices of the intra- and extrasynovial arterial flow (spectral Doppler). The flow pattern was estimated by the indices of pulsatility (PI) and resistance (RI). RESULTS: The sonographic measurements of joint space were reproducible with intraobserver, intraclass correlation coefficients (ICC) 0.82-0.97 (p<0.0001) and interobserver ICC 0.81 (p<0.0001). The mean (SD) fraction of the synovium vascularised in the patients was 0.15 (0.15). The synovial blood flow was characterised by a diastolic flow-that is, the flow persisting during the diastole. The mean (SD) PI was 1.92 (1.18) and RI 0.70 (0.13). The estimated vascular fraction correlated with the erythrocyte sedimentation rate (ESR) (r(s)=0.53, p=0.03). The relative Pi (rPi), an estimate of an abnormally low resistance to vascularisation, correlated with both ESR (r(s)=-0.557, p<0.05) and Health Assessment Questionnaire score (r(s)=-0.584, p<0.05). After an injection of contrast Levovist the vascular fraction increased, while no difference in PI and RI was observed. CONCLUSION: Ultrasonography is a reliable tool for estimating the size of the joint space and the synovial activity measured by the degree of vascularisation and pattern of flow. Ultrasonography may be useful in monitoring the synovial inflammation in rheumatoid arthritis.

Adult↗

Signal analysis of behavioral and molecular cycles.

BACKGROUND: Circadian clocks are biological oscillators that regulate molecular, physiological, and behavioral rhythms in a wide variety of organisms. While behavioral rhythms are typically monitored over many cycles, a similar approach to molecular rhythms was not possible until recently; the advent of real-time analysis using transgenic reporters now permits the observations of molecular rhythms over many cycles as well. This development suggests that new details about the relationship between molecular and behavioral rhythms may be revealed. Even so, behavioral and molecular rhythmicity have been analyzed using different methods, making such comparisons difficult to achieve. To address this shortcoming, among others, we developed a set of integrated analytical tools to unify the analysis of biological rhythms across modalities. RESULTS: We demonstrate an adaptation of digital signal analysis that allows similar treatment of both behavioral and molecular data from our studies of Drosophila. For both types of data, we apply digital filters to extract and clarify details of interest; we employ methods of autocorrelation and spectral analysis to assess rhythmicity and estimate the period; we evaluate phase shifts using crosscorrelation; and we use circular statistics to extract information about phase. CONCLUSION: Using data generated by our investigation of rhythms in Drosophila we demonstrate how a unique aggregation of analytical tools may be used to analyze and compare behavioral and molecular rhythms. These methods are shown to be versatile and will also be adaptable to further experiments, owing in part to the non-proprietary nature of the code we have developed.

Activity Cycles↗

A versatile route to 3-benzoheteroepines containing group 15 and 16 heavier elements involving several novel ring systems, and their thermal stabilities.

The C-unsubstituted 3-benzoheteroepines (2a-g) containing group 15 (P, As, Sb, and Bi) and group 16 (S, Se, and Te) heavier elements were prepared by the reaction of the corresponding metal reagents with (Z,Z)-o-bis(beta-lithiovinyl)benzene (5) which was derived in two steps from a common o-phthalaldehyde (3). The heteroepines (2) thus obtained were thermally labile towards heteroatom extrusion, and their half-lives on heating estimated from (1)H-NMR spectral analysis showed that the 3-benzoheteroepines (2) were far less stable than the corresponding 1-benzoheteroepines (1). The 2,4-bis(trimethylsilyl)-3-benzoheteroepines (17) containing Sb, Bi, and Te were also prepared from o-diiodobenzene (9) in 6 steps and were found to be more stable than the corresponding C-unsubstituted heteroepines (2).

Chemical Phenomena↗

Fluorescence spectra provide information on the depth of fluorescent lesions in tissue.

The fluorescence spectrum measured from a fluorophore in tissue is affected by the absorption and scattering properties of the tissue, as well as by the measurement geometry. We analyze this effect with Monte Carlo simulations and by measurements on phantoms. The spectral changes can be used to estimate the depth of a fluorescent lesion embedded in the tissue by measurement of the fluorescence signal in different wavelength bands. By taking the ratio between the signals at two wavelengths, we show that it is possible to determine the depth of the lesion. Simulations were performed and validated by measurements on a phantom in the wavelength range 815-930 nm. The depth of a fluorescing layer could be determined with 0.6-mm accuracy down to at least a depth of 10 mm. Monte Carlo simulations were also performed for different tissue types of various composition. The results indicate that depth estimation of a lesion should be possible with 2-3-mm accuracy, with no assumptions made about the optical properties, for a wide range of tissues.

Algorithms↗

Color flow mapping in obstetrics.

Blood flow studies are now used extensively in evaluating fetal peripheral circulation. By using continuous and pulsed Doppler ultrasonic equipment the raw Doppler shifts caused by blood moving in the vessels can be analyzed in different ways. Spectral data can be used to estimate blood velocity and volume in the vessel, and pulsatility characteristics and turbulence. All these data are obtained from very restricted areas within the body so that aiming of the probe is critical, and obtaining data from an area requires long and tedious effort. Color flow mapping which has recently been introduced into clinical practice provides an overview of flow velocities and directions within an area. Color coded flow can be displayed by applying the moving target identification principle (as in radar) to an ultrasonic diagnostic instrument where sound waves are used instead of radio waves. The value of flow mapping has already been recognized in the diagnosis of congenital and acquired heart disease in adults and children. Direct visualization of intracardiac flow provides for a fast and accurate diagnosis of various cardiac defects, such as the ventricular septal defect, without cardiac catheterization. In the present study we have investigated the potential of color flow mapping in obstetrics. 211 pregnant women were examined between the 15th and 40th weeks of pregnancy. 193 of them had normal pregnancies; 18 were abnormal, including 11 cases of insulin dependent diabetes, 2 cases of Rh-immunization, 2 cases of nonimmunologic fetal hydrops, 2 cases of fetal cardiac structural defects and 1 case of intrauterine growth retardation. Intracardiac flow was clearly seen in 78% (35/45) of cases at the gestational age between the 20th and 24th weeks. After the 24th week the heart flow visualization rate gradually decreased to 31% (11/36) before term. In 2 cases of congenital heart abnormality flow mapping enabled clear visualization of reverse flow through the tricuspid valve and diagnosis of valvular insufficiency. The best results in studies of flow in peripheral vessels were obtained from umbilical vessels which were seen in all cases after the 15th week. The flow visualization rate was much lower when the fetal aorta, intrahepatic umbilical vein and internal carotid artery were studied. The comparison between the diameter of the umbilical vein measured on the B-scan and flow width in the same vessel showed no significant difference (t = 0.26; p less than 0.01; N = 209).(ABSTRACT TRUNCATED AT 400 WORDS)

Blood Flow Velocity↗

[X-ray fluorescence analysis of Kaolin concentrate].

An X-ray fluorescence technique was developed for the analysis of Al2O3, SiO2 and impurities in Kaolin concentrate. Two standard series, one for dual series of Al2O3-SiO2 and the other for all of the impurities, were prepared, respectively, from spectral pure agents based on the estimates of analyte content ranges and matrix effects in Kaolin concentrate. Powderd sample after determination of loss of ignition was pressed into disc by means of a little powderd sample preparation method modyfied in this paper. All analyte contents were calculated directly from the calibration curves without any matrix correction. In contrast to fusion technique the method has no the effects of flux impurities and dilute ration on the determination of minor and micro components in Kaolin concentrate. Errors of the results are < 1% for major component Al2O3, SiO2 and total, and < 5% for most and < 10% for few of the impurities.

English Abstract↗

[Optimal structure of the connections of the electrical processes in the human cerebral cortex and its disorders in changes of state].

In this paper the data are analyzed on the human EEG investigation. Significance is shown of parameters of correlative and spectral-coherent EEG functions for the estimation of the brain functional state of healthy people and patients with local cerebral lesions. In the norm, the parameter of the mean coherence is stable, its characteristics correspond to the optimum cortical tone, the most favourable for the performance of the cortical functions. In healthy people unstable, individual, different in different cortical areas changes of the spectrum details, coherence and phases reflect local processes, taking place at the optimum mean level of the coherence and form the cortical mosaic. These two sides of the intercentral relations of the electrical processes (optimum level of coherence and dynamic mosaic of connections of separate rhythms) reflect the most favourable conditions of the nervous processes development. In the brain pathology, different forms are noted of deviations from the system of intercentral relations and levels of coherence of cortical electrical processes.

Brain Diseases↗

An investigation of simulated umbilical artery Doppler waveforms. III. The effect of noise reduction algorithms on the maximum frequency envelope and on the pulsatility index.

The effect of two noise reduction algorithms on the accuracy of estimation of the maximum frequency envelope and pulsatility index (PI) of simulated umbilical artery Doppler waveforms was investigated. The algorithms were: first, smoothing of the envelope from unfiltered Doppler spectra using a double window modified trimmed mean (DWMTM) filter and second, speckle and noise reduction of the Doppler spectrum using an image processing method. The test population consisted of waveforms were the degree of beam-vessel misalignment had been varied. The accuracy of estimation of the maximum frequency envelope and the PI was calculated by comparing each set of waveforms with the gold-standard maximum frequency envelope from the ensemble averaged waveform obtained with no misalignment. Speckle reduction gave rise to PI values that were low by approximately 0.1 (3%-4%). When there was no background noise present the improvements in envelope estimation were factors of 1.27 and 1.24, respectively, for the DWMTM method and the spectral filter, whereas the factors were 1.56 and 2.07 when background noise was present. For estimation of PI the DWMTM filter was superior. For no background noise the DWMTM filter gave a factor of 3.36 improvement whereas there was no improvement with the spectral filter. When background noise was present the factors for improvement in PI estimation were 2.39 and 4.16.

Algorithms↗

Photopigment basis for dichromatic color vision in the horse.

Horses, like other ungulates, are active in the day, at dusk, dawn, and night; and, they have eyes designed to have both high sensitivity for vision in dim light and good visual acuity under higher light levels (Walls, 1942). Typically, daytime activity is associated with the presence of multiple cone classes and color-vision capacity (Jacobs, 1993). Previous studies in other ungulates, such as pigs, goats, cows, sheep and deer, have shown that they have two spectrally different cone types, and hence, at least the photopigment basis for dichromatic color vision (Neitz & Jacobs, 1989; Jacobs, Deegan II, Neitz, Murphy, Miller, & Marchinton, 1994; Jacobs, Deegan II, & Neitz, 1998). Here, electroretinogram flicker photometry was used to measure the spectral sensitivities of the cones in the domestic horse (Equus caballus). Two distinct spectral mechanisms were identified and are consistent with the presence of a short-wavelength-sensitive (S) and a middle-to-long-wavelength-sensitive (M/L) cone. The spectral sensitivity of the S cone was estimated to have a peak of 428 nm, while the M/L cone had a peak of 539 nm. These two cone types would provide the basis for dichromatic color vision consistent with recent results from behavioral testing of horses (Macuda & Timney, 1999; Macuda & Timney, 2000; Timney & Macuda, 2001). The spectral peak of the M/L cone photopigment measured here, in vivo, is similar to that obtained when the gene was sequenced, cloned, and expressed in vitro (Yokoyama & Radlwimmer, 1999). Of the ungulates that have been studied to date, all have the photopigment basis for dichromatic color vision; however, they differ considerably from one another in the spectral tuning of their cone pigments. These differences may represent adaptations to the different visual requirements of different species.

Adaptation, Physiological↗

Rank-shaping regularization of exponential spectral analysis for application to functional parametric mapping.

Compartmental models are widely used for the mathematical modelling of dynamic studies acquired with positron emission tomography (PET). The numerical problem involves the estimation of a sum of decaying real exponentials convolved with an input function. In exponential spectral analysis (SA), the nonlinear estimation of the exponential functions is replaced by the linear estimation of the coefficients of a predefined set of exponential basis functions. This set-up guarantees fast estimation and attainment of the global optimum. SA, however, is hampered by high sensitivity to noise and, because of the positivity constraints implemented in the algorithm, cannot be extended to reference region modelling. In this paper, SA limitations are addressed by a new rank-shaping (RS) estimator that defines an appropriate regularization over an unconstrained least-squares solution obtained through singular value decomposition of the exponential base. Shrinkage parameters are conditioned on the expected signal-to-noise ratio. Through application to simulated and real datasets, it is shown that RS ameliorates and extends SA properties in the case of the production of functional parametric maps from PET studies.

Algorithms↗

Sequential characterization of atrial tachyarrhythmias based on ECG time-frequency analysis.

A new method for characterization of atrial arrhythmias is presented which is based on the time-frequency distribution of an atrial electrocardiographic signal. A set of parameters are derived which describe fundamental frequency, amplitude, shape, and signal-to-noise ratio. The method uses frequency-shifting of an adaptively updated spectral profile, representing the shape of the atrial waveforms, in order to match each new spectrum of the distribution. The method tracks how well the spectral profile fits each spectrum as well as if a valid atrial signal is present. The results are based on the analysis of a learning database with signals from 40 subjects, of which 24 have atrial arrhythmias, and an evaluation database with 211 patients diagnosed with atrial fibrillation. It is shown that the method robustly estimates fibrillation frequency and amplitude and produces spectral profiles with narrower peaks and more discernible harmonics when compared to the conventional power spectrum. The results suggest that a rather strong correlation exist between atrial fibrillation frequency and f wave shape. The developed set of parameters may be used as a basis for automated classification of different atrial rhythms.

Algorithms↗