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Postnatal development of EEG patterns, catecholamine contents and myelination, and effect of hyperthyroidism in Suncus brain.

The postnatal development of the central nervous system (CNS) in house musk shrew in the early stage of maturation was studied. The electroencephalogram (EEG) and visual evoked potential (VEP) in association with catecholamine contents and myelin basic protein (MBP) immunoreactivity were carried out from the 1st to the 20th day of postnatal age. Different EEG patterns which were specific to behavioral states (awake and drowsy) were first recorded on the 5th day, and the total power which was obtained by power spectrum analysis increased after this stage. The latencies of all peaks in VEP markedly shortened between the 5th and the 7th day. Noradrenalin (NA) content of the brain showed a slight increase after the 3rd day, and reached maximum levels on the 7th day, which was delayed a few days compared to dopamine (DA). In hyperthyroidism, the peak latency of VEP was shortened and biosynthesis of NA in cerebral cortex and DA in hippocampus was accelerated. The most obvious change in MBP-immunoreactivity of the telencephalon occurred from the 7th to the 10th day. These morphological changes in the brain advanced at the identical time-course to those in the electrophysiological development and increment of DA and NA contents.

Aging↗

Isolation rearing affects sequential organization of motor behavior in post-pubertal but not pre-pubertal Lister and Sprague-Dawley rats.

OBJECTIVE: Dopaminergic and serotonergic modulation potently influences the sequential organization of rat movements in a simple unconditioned motor paradigm. Rats reared in social isolation post-weaning differ profoundly from their socially reared litter mates on behavioral, neurochemical, and neuroanatomical measures. This investigation examined (1) whether social isolation significantly affects the sequential organization of rat movements, (2) whether these changes occur at different ages, and (3) whether these changes differ across strains. METHOD: male Lister and Sprague Dawley rats reared in isolation post-weaning and socially reared controls were tested at 2 and 8 weeks post-weaning, in the Behavioral Pattern Monitor for 30-min sessions. The amount of activity and the spatial patterns of movements as measured by both the spatial scaling exponent and the fluctuation spectrum of local spatial scaling exponents were assessed in 10-min intervals. RESULTS: Habituation of locomotor activity was significantly attenuated in isolation reared rats during the 30-min sessions irrespective of strain. Spatial patterns of movements were significantly affected by isolation rearing in movements in post-pubertal but not pre-pubertal Lister and Sprague-Dawley rats. The spatial scaling exponent and the fluctuation spectrum analysis revealed a shift towards straight, distance-covering, and repetitive movements rather than a complex re-organization of the behavioral repertoire. CONCLUSIONS: Isolation rearing profoundly affects the sequential organization of movements in post-pubertal rats, suggesting that emerging behavioral dysfunctions parallel developmentally those found in patients with schizophrenia.

Animals↗

Is heart rate variability a reliable method to assess autonomic modulation in left ventricular dysfunction and heart failure? Assessment of autonomic modulation with heart rate variability.

Autonomic dysfunction seems to be involved in the progression and prognosis of congestive heart failure. Measurement of heart rate variability (HRV) provides a noninvasive method to obtain reliable and reproducible information on autonomic modulation of heart rate, but there is a difficulty in using HRV as a quantitative estimate of autonomic dysfunction in heart failure. This study was aimed at testing the hypothesis that abnormal modulation of heart rate assessed by power spectrum analysis may be present also in asymptomatic patients with left ventricular dysfunction and progress in patients with overt symptoms of congestive heart failure. HRV was measured in three groups of subjects: Group 1: 30 patients with chronic heart failure; Group 2: 21 patients with asymptomatic left ventricular dysfunction; and Group 3: 25 healthy volunteers as control group. HRV was evaluated by autoregressive spectral analysis with 600-beat ECG samples, while subjects were quietly recumbent (BSI), in conditions of controlled breathing (15 acts/min) (RSC) and passive orthostatism after tilting (80 degrees) (TLT). Patients in group 1 showed a reduction in the standard deviation of the R-R intervals (SDRR) (p<0.0003) and in the low frequency component (LF) (p<0.0001) compared to normal subjects. Low frequency component was not detectable in 11 patients of group I (p<0.0008). On RSC and TLT, group 1 failed to show any modification in the low frequency and high frequency components (HF) under any stimulation. Group 2 showed no modification at baseline evaluation, no increase in the high frequency component on RSC and in LF during TLT compared to controls (p<0.01 and p<0.0001 respectively). At baseline, group 1 had a lower SDRR (p<0.03) and LF (p<0.0001) vs. group 2, whereas during stimulation the two groups exhibited the same behaviour. In conclusion, reduced heart rate variability is specific for both asymptomatic and symptomatic post-ischemic left ventricular dysfunction. Our results suggest that frequency domain analysis of heart rate variability during a stimulation test allows a more accurate definition of the degree of autonomic control of heart rate.

Autonomic Nervous System↗

New biochip technology for label-free detection of pathogens and their toxins.

microSERS is a new biochip technology that uses surface-enhanced Raman scattering (SERS) microscopy for label-free transduction. The biochip itself comprises pixels of capture biomolecules immobilized on a SERS-active metal surface. Once the biochip has been exposed to the sample and the capture biomolecules have selectively bound their ligands, a Raman microscope is used to collect SERS fingerprints from the pixels on the chip. SERS, like other whole-organism fingerprinting techniques, is very specific. Our initial studies have shown that the Gram-positive Listeria and Gram-negative Legionella bacteria, Bacillus spores and Cryptosporidium oocysts can often be identified at the subspecies/strain level on the basis of SERS fingerprints collected from single organisms. Therefore, pathogens can be individually identified by microSERS, even when organisms that cross-react with the capture biomolecules are present in a sample. Moreover, the SERS fingerprint reflects the physiological state of a bacterial cell, e.g., when pathogenic Listeria and Legionella were cultured under conditions known to affect virulence, their SERS fingerprints changed significantly. Similarly, nonviable (e.g., heat- or UV-killed) microorganisms could be differentiated from their viable counterparts by SERS fingerprinting. Finally, microSERS is also capable of the sensitive and highly specific detection of toxins. Toxins that comprised as little as 0.02% by weight of the biomolecule-toxin complex produced strong, unique fingerprints when spectra collected from the complexes were subtracted from the spectra of the uncomplexed biomolecules. For example, aflatoxins B(1) and G(1) could be detected and individually identified when biochips bearing pixels of antibody or enzyme capture biomolecules were incubated in samples containing one or both aflatoxins, and the spectra were then collected for 20 s from an area of the biomolecule pixel approximately 1 microm in diameter. In the future, we plan to investigate the use of hyperspectral imaging Raman microscopy for collecting fingerprints from all the pixels on the biochip, individually yet simultaneously, to enable the rapid detection of diverse pathogens and their toxins in a sample, using a single biochip.

Aflatoxins↗

Postural sway and diabetic peripheral neuropathy.

The focus of this paper is to evaluate the relationship between postural sway assessed by a gravicorder and peripheral neuropathy in patients with type 2 diabetes. Posturography (GRAVICHART), electrophysiological tests, and power spectrum analysis of heart rate fluctuations were performed in the following age-matched subjects: 123 type 2 diabetic patients without peripheral neuropathy, 32 type 2 diabetic patients with peripheral neuropathy, and 55 healthy control subjects. All diabetic patients with a history of clinical neurological dysfunction were excluded from the study. Significant correlations were found between parameters of GRAVICHART and some parameters of the electrophysiological tests as well as the parameters of heart rate variability. The envelope area and the length per time were larger in type 2 diabetic patients with peripheral neuropathy than in patients without peripheral neuropathy and the control subjects. There were no significant differences in the GRAVICHART parameters between diabetic patients without neuropathy and the healthy control subjects. Type 2 diabetic patients with peripheral neuropathy exhibited an inability to maintain an upright posture. The GRAVICHART can indicate the early stage of postural balance impairment.

Albuminuria↗

Genotoxicity and toxicity assay of water sampled from a radium production industry storage cell territory by means of Allium-test.

Water from natural reservoirs located near the radium production industry storage cell were analyzed using the anaphase-telophase chromosome aberration assay that was carried out on Allium schoenoprasum L. meristematic root tip cells. (262)Ra, (228)U, (232)Th, (210)Pb and (210)Po concentrations in all samples were found not to exceed the radioactivity concentration guides. The concentrations of 10 heavy metal ions were measured in water samples, but only Zn and Mn levels exceeded the maximum permissible concentration for the natural reservoirs. All water samples caused a significant increase of the chromosome aberration frequency as compared to control. The chromosome aberration spectrum analysis shows that the genotoxic effect was a result of chemical toxicity mainly. Two samples from the brook springhead were found to be toxic. The regression analysis results show that the mitotic index increased in parallel to Zn ion levels, and decreased with higher (238)U concentrations. The water samples genotoxicity positively correlated with the Zn concentration. The present work demonstrates that in order to achieve pollutant screening, it is not sufficient to determine the pollutants concentration only. Adequate conclusions on the risk due to environment contamination need to be based on the additional simultaneous use of toxicity and genotoxicity tests. When bioassays indicate some genotoxic and toxic effects, the determination of the chemical composition of the samples is then required. A combination of these two methods allows the identification of the elements that require constant biological monitoring. In the study reported here, those elements are Zn and (238)U.

Allium↗

Preparation of a dansylated fibrate, a new fluorescent tool to study peroxisome proliferation. Effect on hepatic-derived cell lines.

The synthesis of a dansylated fibrate (DNS-X) has been performed in order to identify the cellular affinity sites of peroxisome proliferators and to establish the subcellular localization of such molecules. DNS-X has been obtained by coupling the dansy1 chloride with the amine resulting from the bezafibrate alkaline hydrolysis. The purified DNS-X has been further characterized by spectrum analysis (UV-Vis, fluorescence, [1H]/[13C]-NMR and mass). At 250 microM and incubated for 48 h with the rat hepatic derived cells (Fao cells), DNS-X stimulates 12-fold the palmitoyl-CoA oxidase, a peroxisome proliferation marker enzyme. This increase is comparable to the one obtained with well known peroxisome proliferators such as bezafibrate or ciprofibrate. The stimulation by DNS-X is specific for the overall molecule since neither the dansyl chloride, the amine, nor the precursors of DNS-X are active. The increase of palmitoyl-CoA oxidase activity is correlated with the increase of the enzyme amount as shown by immunoblotting. In agreement with the species-specificity of the fibrate neither DNS-X, bezafibrate nor ciprofibrate significantly increase palmitoyl-CoA oxidase activity and the enzyme amount in human hepatic-derived cells, HepG2. This work shows that the dansylated fibrate is a new fluorescent tool to study the subcellular localization and identification of high affinity binding sites, then further on, to elucidate the peroxisome proliferation mechanism and the action of hypolipidaemic agents of the fibrate family.

Animals↗

Spectral parameter imaging for detection of prognostically significant histologic features in uveal melanoma.

Specific extracellular matrix patterns in uveal melanoma are associated with metastatic risk. The laminin-rich composition and dimensions (on the order of a wavelength or less) of these structures suggest that acoustic backscatter might be affected by their presence. In this study, 10-MHz radiofrequency (RF) ultrasound (US) data were acquired before surgical removal of 117 eyes with uveal malignant melanoma. Histologic sections were evaluated for the presence of matrix patterns and acoustic backscatter was characterized using calibrated spectrum analysis. Statistical correlations between acoustic and histologic patterns were determined and linear discriminant analysis (LDA) and radial basis networks (RBN) were used to develop classification models for histologically based risk groups. Statistically significant correlations were found between acoustic parameters and the presence of histologic matrix-rich patterns. Retrospective classification accuracies of 74.4% and 78.6% were obtained with LDA and RBN, respectively. Leave-one-out analyses indicated estimated predictive accuracies of 71.8% and 75.0% for LDA and RBN, respectively.

Extracellular Matrix↗

Evaluation of normal and abnormal hyoid bone movement during swallowing by use of ultrasound duplex-Doppler imaging.

We developed a new method to analyze normal and abnormal movements of the hyoid muscular region as an indicator of hyoid bone motion during swallowing using ultrasound duplex-Doppler imaging. Hyoid bone motion can be monitored by studying the Doppler shift spectra and B-mode images produced by ultrasound duplex imaging of the hyoid region muscular attachments. We can accurately determine swallowing duration and trajectory of hyoid bone movement. This procedure can assist in discriminating between normal and abnormal movements of the hyoid bone and the surrounding muscles during swallowing. This method appears to be a highly consistent measure. We suggest that Doppler spectrum analysis can be used for defining hyoid position and displaying accurate movement, which may be useful in the diagnosis of swallowing disorders.

Adult↗

Correlation of ultrasound parameter imaging with microcirculatory patterns in uveal melanomas.

Previous studies demonstrated a correlation between acoustic backscatter parameters and survival in ocular melanoma. The histologic presence of microvascular networks in ocular melanoma is also associated with death from metastases. This study tests the hypothesis that melanomas grouped on the basis of these microvascular patterns are separable by ultrasound spectrum analysis. We scanned 40 melanomas using a 10-MHz ultrasound unit equipped for digitization of radio frequency data. After enucleation, tumors were sectioned in planes corresponding to the ultrasonographic examination and stained to demonstrate microcirculation. Acoustic spectral parameters were compared between 14 melanomas with a nevuslike microcirculation and 26 with foci of high-risk microvascular structures. Smaller scatterer size, lower acoustic concentration and greater spatial variability were found to correlate with high-risk microvascular patterns and areas of cystic degeneration. We suggest that nonvascular extracellular matrix components associated with microvessels may be responsible for the correlation of acoustic parameters with microvascular pattern and distribution.

Choroid Neoplasms↗

Statistical framework for ultrasonic spectral parameter imaging.

This study examines the statistics of ultrasonic spectral parameter images that are being used to evaluate tissue microstructure in several organs. The parameters are derived from sliding-window spectrum analysis of radiofrequency echo signals. Calibrated spectra are expressed in dB and analyzed with linear regression procedures to compute spectral slope, intercept and midband fit, which is directly related to integrated backscatter. Local values of each parameter are quantitatively depicted in gray-scale cross-sectional images to determine tissue type, response to therapy and physical scatterer properties. In this report, we treat the statistics of each type of parameter image for statistically homogeneous scatterers. Probability density functions are derived for each parameter, and theoretical results are compared with corresponding histograms clinically measured in homogeneous tissue segments in the liver and prostate. Excellent agreement was found between theoretical density functions and data histograms for homogeneous tissue segments. Departures from theory are observed in heterogeneous tissue segments. The results demonstrate how the statistics of each spectral parameter and integrated backscatter are related to system and analysis parameters. These results are now being used to guide the design of system and analysis parameters, to improve assays of tissue heterogeneity and to evaluate the precision of estimating features associated with effective scatterer sizes and concentrations.

Carcinoma↗

Synergistic sedative effects of noradrenergic alpha(1)- and beta-receptor blockade on forebrain electroencephalographic and behavioral indices.

The locus coeruleus-noradrenergic system exerts an activating influence on forebrain neuronal and behavioral activity states. For example, in the anesthetized rat, unilateral locus coeruleus stimulation elicits bilateral activation of forebrain electroencephalographic activity. Pretreatment with a noradrenergic beta-antagonist blocks this effect, suggesting that beta-receptors play a critical role in locus coeruleus-dependent activation of the forebrain. Consistent with this, stimulation of beta-receptors located in certain basal forebrain structures evokes sustained periods of alert waking in the unanesthetized rat. Similar forebrain and behavioral activating effects are observed with alpha(1)-receptor stimulation within these basal forebrain regions. To assess the extent to which alpha(1)- and beta-receptors contribute to the maintenance of behavioral and forebrain activation, we examined the electroencephalographic and behavioral effects of alpha(1)-, beta- and combined alpha(1)/beta-receptor blockade in the unanesthetized rat. Rats were treated individually or in combination with either varying doses of the alpha(1)-antagonist, prazosin (intraperitoneally), and/or the beta-antagonist, timolol (intracerebroventricularly). Thirty minutes following treatment, animals were placed in a mildly-arousing novel environment, which has been demonstrated previously to elicit activation of central noradrenergic systems and sustained waking in vehicle-treated controls. Behavior and electroencephalographic activity were recorded and later scored. Electroencephalographic activity was analysed using power spectrum analysis. The following were observed: (i) beta-receptor blockade alone does not alter behavioral or electroencephalographic indices of alert waking; (ii) alpha(1)-receptor blockade alone increases high-voltage spindle activity in cortical electroencephalographic activity that was associated with decreased behavioral activity; (iii) combined alpha(1)- and beta-receptor blockade elicits a substantial increase in slow-wave activity (0.33-2.0Hz), also in association with decreased behavioral activity. All of these effects were dependent on the dose administered and time following initiation of testing. These results indicate that the combined actions of alpha(1)- and beta-receptors exert distinct and synergistic actions on cortical neuronal activity patterns that are essential elements of alert waking.

Adrenergic alpha-Antagonists↗

Manipulations during forced wakefulness have differential impact on sleep architecture, EEG power spectrum, and Fos induction.

We propose a hypothesis suggesting that the most prominent experiences occurring during wakefulness activate specific clusters of neurons related to such experiences. These neurons could possibly then evoke the release of various types of sleep-inducing molecules, thereby causing different patterns of sleep architecture. In this study, we therefore sought to determine whether manipulations of behavior during wakefulness, such as forced wakefulness induced by gentle handling, forced wakefulness associated with a stressful condition such as immobilization, or forced wakefulness associated with excess intake of palatable food, could result in a variation of Fos immunoreactivity in selective brain structures and could also result in different sleep and EEG power density patterns. The results showed that the sleep-wake cycle of rats after all the experimental manipulations was different not only with respect to the control group but also among themselves. Additionally, power spectrum analysis showed an increase of 0.25-4.0 Hz in all experimental manipulations, whereas the 4.25-8.0 Hz increase occurred only in the situation of forced wakefulness plus stress. The Fos induction showed activation of cell clusters in cortical areas and telencephalic centers, in several hypothalamic nuclei, in monoaminergic cell groups, and in brain stem nuclei. The density of Fos-immunoreactive neurons varied in relation to the different paradigms of forced wakefulness. These results suggest that activation of cell clusters in the brain are related to the type of manipulation imposed on the rat during wakefulness and that such variation in cell activation prior to sleep may be associated with sleep architecture and EEG power.

Animals↗

Periodic spectral components of fetal heart rate variability reflect the changes in cord arterial base deficit values: a preliminary report.

Fetal distress changes the function of the autonomic nervous system. These changes are reflected in the fetal heart rate and can be quantified with power spectrum analysis of heart rate variability. The purpose of this study was to find out whether spectral components of fetal heart rate variability (FHRV) during labor are associated with fetal cord arterial base deficit values at birth. The association between FHRV and umbilical cord arterial base deficit was studied in 14 singleton fetuses with normal pregnancy at 35-40 weeks of gestation. Fetal ECG was recorded by scalp-electrode using a STAN Fetal ECG monitor (Cinventa Ab, Mölndal, Sweden). FHRV was quantified by computing Fast-Fourier-transformed heart rate (HR) spectra at three frequency bands: low-frequency (LF) 0.03-0.07 Hz, mid-frequency (MF) 0.07-0.13 Hz and high-frequency (HF) 0.13-1.0 Hz. We found that total FHRV and MF FHRV were lower in fetuses with cord arterial base deficit 8 to 12 mmol/L in comparison to the fetuses with normal cord arterial base deficit value (P=0.02 and P=0.01, respectively). A linear correlation was found between the spectral densities and the cord arterial base deficit values (r=0.4 and r=0.6, respectively). We conclude that the results suggest changes in the autonomic nervous cardiac control in fetuses with cord arterial base deficit between 8 to 12 mmol/L. The clinical applicability of our observations on FHRV in predicting fetal distress remains to be further studied.

Carbon Dioxide↗

Application of a new high-performance liquid chromatographic method for measuring selected polyphenols in human plasma.

We developed a method to measure plasma levels of selected polyphenols before and after ingestion of green tea. Blood samples were obtained from four healthy women before and 30 and 50 min after the ingestion of 300 ml of green tea infusion. A 1-ml volume of plasma was hydrolysed with 0.5 M HCl-methanol (1:1, v/v) for 30 min at room temperature, extracted with ethyl acetate and separated by reversed-phase chromatography. Polyphenols were identified on the basis of their retention times and by spectrum analysis. Green tea caffeine has the same retention times as caffeic acid. Consumption of green tea produces a notable increase in the plasma levels of caffeine plus caffeic acid and the appearance of measurable levels of epigallocatechingallate. In conclusion, the method was found to have the requisite features of specificity and sensitivity for monitoring plasma levels of selected tea polyphenols.

Chromatography, High Pressure Liquid↗

Acoustically and electrically evoked contralateral suppression of otoacoustic emissions in guinea pigs.

It is generally accepted that stimulation of the efferent auditory system results in changes of cochlear activity. A simple method of activating the olivocochlear pathway by contralateral electrical stimulation of the round window (ES-RW) was used in this study with the aim of comparing the efficacy of acoustically and/or electrically evoked contralateral suppression. The suppression of transient evoked otoacoustic emissions (TEOAEs) and distortion product otoacoustic emissions (DPOAEs) was elicited by contralateral acoustic stimulation (AS) (61 dB SPL continuous white noise), and/or by electrical stimulation of an electrode implanted at the contralateral round window (monopolar rectangular pulses 0.1 ms, repetition rate 300 Hz, intensity 50-100 PA) in 12 guinea pigs. The average value of contralateral suppression of TEOAEs amounted to 1.04 +/- 0.48 dB for acoustic stimulation and 0.97 +/- 0.53 dB for round window electrical stimulation. The simultaneous presentation of both acoustic and electrical stimulation had only a slight additive effect and resulted in 1.27 +/- 0.79 dB diminution of TEOAEs. The suppression of DPOAEs during contralateral acoustic and electrical stimulation was evident mainly at low and middle frequencies (14 kHz). In two guinea pigs the maximum DPOAE suppression was present at high frequencies. The average values of contralateral suppression measured at individual f2 frequencies of DPOAEs were similar to those calculated from 1/4 octave power spectrum analysis of the TEOAEs in half of the animals. The results demonstrated that contralateral ES-RW had a similar suppressive effect on TEOAEs and DPOAEs as did contralateral AS and simultaneous AS+(ES-RW). The results of spectral analysis suggested that both modes of contralateral stimulation excited similar sensory cochlear elements and induce comparable suppression of both TEOAEs and DPOAEs.

Acoustic Stimulation↗

Atomic emission spectrometric determination of ephedrine, cinchonine, chlorpheniramine, atropine and diphenhydramine based on formation of ion associates with ammonium reineckate.

Ion-associate complexes of ephedrine HCl (I), cinchonine HCl (II), chlorpheniramine maleate (III), atropine sulphate (IV) and diphenhydramine HCl (V) with ammonium reineckate were precipitated and their solubilities were studied as a function of pH, ionic strength and temperature. Saturated solutions of each ion-associate under the optimum precipitation conditions were prepared and the Cr ion content in the supernatant was determined. The solubility products were thus elucidated at different temperatures. A new accurate and precise method using direct current plasma-atomic emission spectrometry for the determination of the investigated drugs in pure solutions and in pharmaceutical preparations is described. The drugs can determined by the present method in the ranges 1.6-52,2.64-85.8,3.12-101.4,5.52-180.4 and 2.72-75.85 microg/ml solutions of I, II, III, IV and V, respectively.

Atropine↗