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PubMed2000

A second photochromic bacteriophytochrome from Synechocystis sp. PCC 6803: spectral analysis and down-regulation by light.

It now appears that photosynthetic prokaryotes and lower eukaryotes possess higher plant phytochrome-like proteins. In this work, a second phytochrome-like gene was isolated, in addition to the recently identified Cph1 phytochrome, from the Synechocystis sp. PCC 6803, and its gene product was characterized photochemically. The open reading frame sll0821 (designated cph2 in this work) has structural characteristics similar to those of the plant phytochromes and the Synechocystis Cph1 with high amino acid sequence homology in the N-terminal chromophore binding domain. The predicted Cph2 protein consists of 1276 amino acids with a calculated molecular mass of 145 kDa. Interestingly, the Cph2 protein has two putative chromophore binding domains, one around Cys-129 and the other around Cys-1022. The Cph2 was overexpressed in E. coli as an Intein/CBD (chitin binding domain) fusion and in vitro reconstituted with phycocyanobilin (PCB) or phytochromobilin (PPhiB). Both the Cph2-PCB and Cph2-PPhiB adducts showed the typical photochromic reversibility with the difference spectral maxima at 643/690 and 655/701 nm, respectively. The Cys-129 was confirmed to be the chromophore binding residue by in vitro mutagenesis and Zn(2+) fluorescence. The microenvironment of the chromophore in Cph2 seems to be similar to that in plant phytochromes. The cph2 gene expression was dark-induced and down-regulated to a basal level by light, like the cph1 gene. These observations suggest that Synechocystis species have multiple photosensory proteins, probably with distinct roles, as in higher plants.

Amino Acid Sequence↗
PubMedDate not supplied

Spectral analysis of low-frequency, active-head vestibulo-ocular reflex responses.

Useful medical diagnostic information has been reported from low-frequency rotational testing of the horizontal vestibulo-ocular reflex (VOR) of patients with vestibular disorders. Servocontrolled rotating systems have been used as the only practical method to generate stimuli over lower VOR frequency response ranges, the decade from 0.01 to 0.1 Hz. Active head movements have been used for testing the human VOR at higher frequencies, exceeding 0.5 Hz. We examined whether active head movements could be used also to test the VORs of subjects over lower frequency ranges, extending to 0.02 Hz. We used a swept-frequency, active head movement protocol to generate a broad-band stimulus. Eye position was recorded with electro-oculography. Head velocity was recorded with a rotational sensor attached to a head band. Six individual test epochs from human subjects were concatenated to form complex, periodic waveforms of head and eye velocity, 75 seconds in duration. Broad-band cross-spectral signal processing methods were used to compute horizontal VOR system characteristics from these waveforms extending from 0.02 to 2 Hz. The low-frequency VOR data appeared to originate from amplitude modulation of high-frequency active movements, acting as carrier signals. Control experiments and processing of simulated data from a known system excluded the possibility of signal processing artifacts. Results from six healthy subjects showed low-frequency gains and phase values in ranges similar to those from published rotational chair studies of normal subjects. We conclude that it is feasible to test the human VOR over extended low-frequency ranges using active head movements because of amplitude modulation of the head and eye signals.

Adult↗
PubMed2003

Spectral analysis of pigment photobleaching in photosynthetic antenna complex LHCIIb.

Light-induced photooxidation of chlorophyll (Chl) a, b and xanthophylls was investigated in LHCIIb, the antenna pigment-protein complex of photosystem II. Absorption difference spectra at normal and low temperatures show initially (at less than 25% Chl a decay) a selective bleaching of a red-shifted Chl b with absorption bands at 487 and 655 nm, Chl b (460/650 nm) and Chl a (433/670 nm), which changes to a less selective photooxidation pattern at deeper bleaching stages. Difference absorption spectra and HPLC analyses indicate different photooxidation rates of pigments in the order neoxanthin>Chl a>lutein approximately Chl b. Despite significant pigment loss as monitored with absorption spectra, CD spectra indicate an essentially complete persistence of the protein secondary structure. Fluorescence excitation spectra suggest the conversion of a small fraction of Chl a into pheophytin a which acts as a fluorescence quencher, possibly through temporary charge separation process. The strong features in the electroabsorption (Stark effect) spectra due to chlorophyll b at 655 nm and a xanthophyll at 510 nm, and the spectral changes mentioned above are assigned to Chl molecules located at several binding sites in LHCIIb protein and are discussed in the context of spatial configuration and interactions of pigment molecules.

Apoproteins↗
PubMedDate not supplied

Is the short-term modulation of heart rate in teleost fish physiologically significant? Assessment by spectral analysis techniques.

Heart rate is an important physiological variable in the control of cardiac output, even in fishes, where the importance of stroke volume has been overemphasized. Except for the myxinoids, the fish heart is innervated by cranial nerve X and the nature of this innervation is mainly inhibitory by parasympathetic fibers, although a sympathetic contribution has also been demonstrated. In mammals, cardiac innervation is not only responsible for the control of mean heart rate but it also modulates the beat-to-beat heart rate changes. These beat-to-beat changes are known as heart rate variability (HRV) and appear to be related to fluctuations in respiration and blood pressure. In this paper we demonstrate the link between cardiac innervation and HRV in several species of teleosts because HRV is greatly decreased after vagotomy or atropinization. In contrast, after abolishing the sympathetic influence with propranolol, only slight changes in total HRV are observed, indicating the restricted importance of the adrenergic innervation in determining phasic changes in HRV despite the significant tonic effect which has been demonstrated. Thus, it appears unlikely that the sympathetic influence will be present in any measured spectral component as suggested previously. Furthermore, clear spectral patterns do not always exist and this may be due to the erratic influence of respiration which is clearly faster than heart rate in all fish species studied. This differs from the slow ventilation frequency displayed by many mammalian species that exerts an influence on a beat-to-beat basis (respiratory sinus arrhythmia). Spectral patterns could also be affected by changing levels of circulating catecholamines, although this is still unproved.

Animals↗
PubMed1996

Coarse graining spectral analysis of HR and BP variability in patients with autonomic failure.

We examined heart rate and blood pressure variability (HRV and BPV) during graded tilt (5 min in each position: supine, -10 degrees, 10 degrees, 30 degrees, 60 degrees, -10 degrees, supine) in autonomic failure patients and age-matched controls. Heart rate was not different between patients and controls and increased with tilt (P < 0.001). Total HRV was reduced in patients (P < 0.03). Patients had reduced low-frequency (0-0.15 Hz) HRV and BPV (P < 0.005). With tilt, low-frequency BPV increased in controls, whereas high-frequency (> 0.15 Hz) BPV increased in patients. The slope of the fractal component (beta) for HRV and BPV was not different between patients and controls. HRV-beta increased (1.5-1.9, P < 0.01) with tilt, but BPV-beta (approximately 1.8) was unaffected. Values of beta close to 1 indicate high signal regulatory complexity, and values of beta close to 2 indicate low complexity. HRV and BPV provide clear evidence of impaired sympathetic and parasympathetic autonomic nervous system response to tilt with autonomic failure. The similarity in signal complexity with reduced fractal and harmonic spectral power, in patients compared with controls, suggests unchanged cardiovascular neural input and integration with reduced output in autonomic failure.

Adult↗
PubMed1998

Power spectral analysis of arterial blood pressure after spinal anesthesia.

BACKGROUND: The continuous, on-line and real-time analysis of the power spectrum (CORAPS) of systemic arterial pressure (SAP) and heart rate (HR) signals is one of the current progresses in the development of non-invasive indexes for autonomic nervous system. It might be a useful clinical tool to monitor the progress of preganglionic block of sympathetic nervous system and the homeostatsis of cardiac neuroregulation after spinal anesthesia. The purpose of this study is to examine the relationship between the hypotensive response and the changes of components in CORAPS throughout the course of spinal anesthesia. METHODS: In 10 ASA class I-II patients (aged from 17 to 75 years) scheduled for elective surgery, we analyzed the changes of spectrum of systemic arterial blood pressure signals during spinal anesthesia. Spinal anesthesia was performed with 0.5% bupivacaine (dosage at 13-15 mg). For CORAPS, SAP signals were simultaneously relayed to an analog-digital converter connected to a computer. Power spectral density of the data was computed using a fast Fourier transform. The areas of the spectral peaks within each measurement were calculated as the follows: very low (BVLF, 0.01-0.08 Hz), low (BLF, 0.09-0.15 Hz), and high (BHF, 0.16-0.25 Hz). RESULTS: During the time interval (S2 period) when spinal anesthesia was initiated and 15 min thereafter the power density of low frequency (BLF), (1.2 vs. 2.3), and very low frequency (BVLF) components decreased, (3.4 vs. 7.8), in comparison with that at the baseline level (S1 period). We also found the shift of dominance between low frequency (BLF) and high frequency (BHF) in S2 period. The effects of intrathoracic pressure on the venous return may contribute to the increase of power density of BHF components (1.7 vs. 1.1). We also found that even in patients with stable hemodynamic variability, the decrease of BLF and BVLF components, and the shift of dominance were the same as in patients with unstable hemodynamic variability. CONCLUSIONS: It might prove that the cephalic spread of spinal block abolished the sympathetic tone of the autonomic nervous system. These results validated previous speculations of a sympathetic block during spinal anesthesia. More importantly, patients with stable hemodynamic variability still have profoundly sympathetic block with decreased power density of BLF and BVLF. The CORAPS is a good clinical monitoring tool to evaluate the homeostatsis of cardiac neuroregulation after spinal anesthesia.

Adolescent↗
PubMed1990

Spectral analysis of canine epicardial electrogram. Short-term variations in the frequency content induced by myocardial ischemia.

Power spectra of epicardial electrograms were studied in 13 anesthetized dogs subjected to occlusion of the left anterior descending coronary artery. Electrograms were obtained from a bipolar electrode placed on the epicardial surface of the left ventricle and recorded before and after coronary occlusion. After digitization, power spectra of the first and every 50th subsequent waveform were evaluated and compared with the power spectrum of the average waveform obtained from the baseline recording. In particular, we examined variations in relative power content in three frequency ranges: 150-250 Hz, previously shown to be directly affected by myocardial ischemia; 40-150 Hz, presumably corresponding to the fine notches and slurs on the body surface QRS; and 2-40 Hz, the low-frequency range. Apart from a mild initial rise during the first 50 heart beats, the power in the high-frequency range gradually decreased, reaching 5% of the control value at wave 500. The power in the mid-frequency range showed a monotonous decrease after the occlusion and reached 16% of the control. The power in the low-frequency range showed a gradual buildup to 140% of the control after 500 heart beats. Therefore, ischemia causes a shift of the high-frequency spectral components of the local electrographic waveform to lower frequencies. Our findings and the fact that body surface ECG is produced by spatial summation of local electric potentials over the different regions of the myocardial tissue may explain two previously described effects of acute myocardial ischemia on the body surface ECG complex. First, local loss of spectral components in the range 150-250 Hz may produce a zone of reduced amplitude within a QRS complex band-pass filtered in this range. Second, displacement of power in the frequency domain may suggest an explanation to the increased incidence of visible notches and slurs on the surface QRS complex, characteristic to various myocardial pathologies.

Animals↗
PubMed1985

Power spectral analysis of the surface electromyogram before and after a warning signal in a reaction movement.

The present study was performed to substantiate the premise that frequency components of the surface EMG before a reaction movement change after a warning signal (S1). Twelve subjects extended rapidly the trunk from a bent position in response to a moving signal (S2) 2 sec after S1. The movement was repeated 20 times. Bipolar surface EMG recordings were made from the erector spinal muscle. A minicomputer calculated power spectra of the surface EMGs for 2 sec before S1 and for 2 sec from S1 until S2 in each trial. From each spectrum, the mean frequency (MF) was calculated. EMG itself was examined. Statistical comparisons of differences between the mean values of the MF before and after S1 showed that a significant difference was found in 5 subjects. For the subjects, the mean value of the MF after S1 was a lower value than that of the MF before S1. Increase of EMG power in frequency range from 30 to 50 Hz was observed in the averaged EMG spectrum after S1 in the subjects. Slowing of the surface EMG during the preparatory period from S1 until S2 was often observed in the subjects. These results show that the power spectrum of the surface EMG after S1 shifted to lower frequencies and suggest that the EMG spectral shift is due to the slowing of the surface EMG in association with motor preparation to perform the movement.

Electromyography↗
PubMed2001

Mass spectral analysis of protein-based radicals using DBNBS. Nonradical adduct formation versus spin trapping.

Protein-based radicals generated in the reaction of ferricytochrome c (cyt c) with H(2)O(2) were investigated by electrospray mass spectrometry (ESI-MS) using 3,5-dibromo-4-nitrosobenzenesulfonate (DBNBS). Up to four DBNBS-cyt c adducts were observed in the mass spectra. However, by varying the reaction conditions (0-5 molar equivalents of H(2)O(2) and substituting cyt c with its cyanide adduct which is resistant to peroxidation), noncovalent DBNBS adduct formation was inferred. Nonetheless, optical difference spectra revealed the presence of a small fraction of covalently trapped DBNBS. To probe the nature of the noncovalent DBNBS adducts, the less basic proteins, metmyoglobin (Mb) and alpha-lactalbumin, were substituted for cyt c in the cyt c/H(2)O(2)/DBNBS reaction. A maximum of two DBNBS adducts were observed in the mass spectra of the products of the Mb/H(2)O(2)/DBNBS reactions, whereas no adducts were detected following alpha-lactalbumin/H(2)O(2)/DBNBS incubation, which is consistent with adduct formation via spin trapping only. Titration with DBNBS at pH 2.0 yielded noncovalent DBNBS-cyt c adducts and induced folding of acid-denatured cyt c, as monitored by ESI-MS and optical spectroscopy, respectively. Thus, the noncovalent DBNBS-cyt c mass adducts observed are assigned to ion pair formation occurring between the negatively charged sulfonate group on DBNBS and positively charged surface residues on cyt c. The results reveal the pitfalls inherent in using mass spectral data with negatively charged spin traps such as DBNBS to identify sites of radical formation on basic proteins such as cyt c.

Animals↗
PubMed1990

Extramitochondrial release of hydrogen peroxide from insect and mouse liver mitochondria using the respiratory inhibitors phosphine, myxothiazol, and antimycin and spectral analysis of inhibited cytochromes.

The fumigant insecticide phosphine (PH3) is known to inhibit cytochrome c oxidase in vitro. Inhibition of the respiratory chain at this site has been shown to stimulate the generation of superoxide radicals (O2-), which dismutate to form hydrogen peroxide (H2O2). This study was performed in order to investigate the production of H2O2 by mitochondria isolated from granary weevil (Sitophilus granarius) and mouse liver on exposure to PH3. Other respiratory inhibitors, antimycin, myxothiazol, and rotenone were used with insect mitochondria. Hydrogen peroxide was measured spectrophotometrically using yeast cytochrome c peroxidase as an indicator. Insect and mouse liver mitochondria, utilizing endogenous substrate, both produced H2O2 after inhibition by PH3. Insect organelles released threefold more H2O2 than did mouse organelles, when exposed to PH3. Production of H2O2 by PH3-treated insect mitochondria was increased significantly on addition of the substrate alpha-glycerophosphate. Succinate did not enhance H2O2 production, however, indicating that the H2O2 did not result from the autoxidation of ubiquinone. NAD(+)-linked substrates, malate and pyruvate also had no effect on H2O2 production, suggesting that NADH-dehydrogenase was not the source of H2O2. Data obtained using antimycin and myxothiazol, both of which stimulated the release of H2O2 from insect mitochondria, lead to the conclusion that glycerophosphate dehydrogenase is a source of H2O2. The effect of combining PH3, antimycin, and myxothiazol on cytochrome spectra in insect mitochondria was also recorded. It was observed that PH3 reduces cytochrome c oxidase but none of the other cytochromes in the electron transport chain. There was no movement of electrons to cytochrome b when insect mitochondria are inhibited with PH3. The spectral data show that the inhibitors interact with the respiratory chain in a way that would allow the production of H2O2 from the sites proposed previously.

Animals↗
PubMed1983

Spectral analysis of all-night sleep EEG in healthy adults.

Power and coherence spectra were computed from all-night sleep EEG records in 6 healthy adult subjects. Derivations were from F3, F4, P3, P4, O1, O2, T3, and T4 to the vertex (Cz). Records were conventionally scored into sleep stages. Average power per sleep stage was maximal at frequencies 0.4-6 c/s in stage 4, at 6-10 c/s in either stage 3 or stage 4, at 12-14 c/s in stage 2 and at 14-30 c/s in stage 1. The average power range from highest values in the lowest frequency band to lowest values in the highest frequency band showed marked differences between sleep stages: It was lowest (12-14 dB) in stage 1, followed by stage 2 (20-22 dB), and stage 3 (16-28 dB), and largest in stage 4 (29-32 dB). REM sleep (15-16 sB) was between stage 1 and 2. The waking state showed an average power range of 11-15 dB. Alpha power at 8-10 c/s in occipital and parietal leads was remarkably constant during sleep, i.e. independent of sleep stage. Coherence showed maximal values at 2-8 c/s in REM sleep, at 8-12 c/s in stage 4, at 12-17 c/s in either stage 3 or 4, and at 17-30 c/s again in stage REM. There was significant coherence increase at 2-8 and 17-30 c/s from NREM to REM sleep, most pronounced between parietal to vertex derivations. Overall coherence between both occipital-to-vertex, or between occipital and parietal-to-vertex derivations, was essentially higher than in the other derivations. The results, essentially, give a comprehensive phenomenology of the dynamic spectral structure of all-night sleep EEG. They suggest that the different brain states during sleep (e.g. stage 1 NREM vs. REM) which are associated with different functions (e.g. hypnagogic hallucinations vs. dreams) differ in EEG spectral parameters if coherence is considered. Likewise, they suggest that studies of automatic sleep staging based exclusively on EEG spectral parameters appear promising.

Adult↗
PubMed1997

Spectral analysis of electrograms during ventricular tachycardia in a canine model: relation with epicardial isochronal maps.

The purpose of this study was to assess the capability of magnitude-squared coherence and bicoherence to differentiate monomorphic ventricular tachycardia (MVT) and polymorphic ventricular tachycardia (PVT) in a canine model and to relate these results to the epicardial isochronal maps on a beat-to-beat basis. Unipolar electrograms were simultaneously recorded from the surface of both ventricles with a 127-lead sock electrode array in 12 open-chest anesthetized dogs. The sampling frequency was 500 Hz. Atrioventricular block was induced by formaldehyde injection into the atrioventricular node. The left anterior descending coronary artery was occluded for 60 minutes under ventricular pacing (140 beats/min). During reperfusion, 12 MVT episodes lasting more than 42 seconds were recorded. Left stellate ganglion stimulation induced five PVT episodes lasting more than 42 seconds. Each of these recordings was divided into seven segments of 3,072 points (6.144 seconds). After visual selection, 104 segments were extracted and classified as 73 MVT and 31 PVT segments. Magnitude-squared coherence was estimated as the cross-spectrum from two epicardial signals (on the right and left ventricles, respectively), normalized with the respective autopower spectrum. Bicoherence was estimated as the bispectrum normalized with the autopower spectrum. Magnitude-squared coherence correctly identified 96% of MVT and 81% of PVT segments for a total accuracy of 91%. Bicoherence estimated with the left ventricular lead correctly identified 100% of MVT and 77% of PVT segments with an accuracy of 93%. Beat-to-beat epicardial maps of MVT displayed a cluster of sites of origin close to the reperfusion area, while the sites of origin from beats during PVT were much more dispersed over both ventricles. A strong and significant correlation was found between the number of electrodes with the earliest epicardial activation and coherence (r = .76, P < .0001) and bicoherence (r = .68, P < .0001), respectively. A high and significant correlation was also found between both spectral estimators (r = .74, P < .0001). Coherence and bicoherence discriminated accurately between MVT and PVT. Coherence achieved better results compared with bicoherence. Coherence and bicoherence measurements showed a quantitative relation with the spatial dispersion of the sites of origin. Both spectral techniques seemed powerful enough to be used in the development of implantable devices.

Animals↗
PubMed1999

Power spectral analysis of heart rate variability during hyperinsulinemia in nondiabetic offspring of type 2 diabetic patients: evidence for possible early autonomic dysfunction in insulin-resistant subjects.

Sympathetic activation has been considered as a link between insulin resistance, hyperinsulinemia, and hypertension. However, little is known about the association between insulin sensitivity and autonomic regulation or about the effect of acute hyperinsulinemia on cardiac sympathovagal balance. The aim of this study was to investigate heart rate variability (HRV) during the euglycemic-hyperinsulinemic clamp in nondiabetic offspring of patients with type 2 diabetes. We studied 35 nondiabetic offspring of patients with type 2 diabetes and 19 control subjects. Probands were chosen from a 10-year follow-up study of patients with well-characterized type 2 diabetes according to their fasting C-peptide level (selected from both ends of the distribution) and from control subjects to form three groups: 1) a group including subjects who were offspring of type 2 diabetic patients with low C-peptide levels (deficient insulin secretion group [IS group], n = 17), 2) a group including subjects who were offspring of type 2 diabetic patients with high C-peptide levels (insulin-resistant group [IR group], n = 18), and 3) a control group without a history of type 2 diabetes in first-degree relatives (n = 19). HRV was assessed at baseline and at the steady state during the euglycemic-hyperinsulinemic clamp. Rates of whole-body glucose uptake (M value) were lower in the IR group than in the IS group and the control group (41+/-3 vs. 54+/-2 vs. 60+/-4 micromol x kg(-1) x min(-1), P < 0.01 and P < 0.01, respectively). In all groups, heart rate increased significantly during hyperinsulinemia. In the IR group, insulin infusion increased total power of HRV [from 7.70+/-0.15 to 8.05+/-0.15 ln(ms2), P < 0.01] and the low frequency-to-high frequency ratio (from 0.62+/-0.14 to 1.14+/-0.18, P < 0.01) and decreased power of the high frequency spectral component (from 5.73+/-0.17 to 5.43+/-0.16 ln(ms2), P < 0.05), whereas in other groups, changes in HRV were not significant. We conclude that the HRV response to acute hyperinsulinemia in the offspring of type 2 diabetic probands was likely to be modulated by the type 2 diabetic phenotype of the parent. In insulin-resistant subjects, autonomic dysfunction may be an earlier defect than hitherto acknowledged.

Adult↗
PubMed1999

Rapid screening of protein profiles of human breast cancer cell lines using non-porous reversed-phase high performance liquid chromatography separation with matrix-assisted laser desorption/ionization time-of-flight mass spectral analysis.

Non-porous reversed-phase (NP-RP) HPLC has been used to rapidly generate protein profiles of whole cell lysates of human breast cancer cell lines. The non-porous packing material used was silica coated with C18, which provided rapid separation with high collection efficiency of proteins from cell lysates. This method was used to study the differences in protein profiles among normal cells and fully malignant cells that share a common genetic background. The highly expressed proteins in each cell type were separated and collected in the liquid state where they were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) to obtain the molecular weight of the proteins. The protein fractions were then subjected to tryptic digestion and analyzed by pulsed delay extraction (PDE)-MALDI-TOFMS to obtain the peptide maps. The expressed proteins were identified based upon the molecular weight and peptide map using database-searching procedures. It is shown that key cancer-related proteins can be detected and identified which may be potentially used as biomarkers for cancer detection.

Biomarkers, Tumor↗
PubMed1995

Spectral analysis of magnetic fields from domestic appliances and corresponding induced current densities in an anatomically based model of the human head.

Magnetic fields emitted by electric appliances such as razors, hair dryers, and drills were measured in the frequency domain. Results show the presence of high-frequency components (up to 96 kHz for razors, up to 3.4 kHz for hair dryers, and up to 8.6 kHz for drills) in the harmonic content of the fields. The measured fields were used to calculate the induced current densities in an anatomically based model of the human head (resolution 1.31 cm) by using the impedance method. The harmonic field contribution to the current density was higher than that from the carrier frequency for all the tested appliances.

Electronics↗