Effects of the static and ELF magnetic fields on the neuronal population activity in Morimus funereus (Coleoptera, Cerambycidae) antennal lobe revealed by wavelet analysis.

To study the influence of a static magnetic field (SMF, 2 mT) and extremely low frequency magnetic field (ELF MF, 50 Hz, 2 mT) on the neuronal population activity, the experiments were performed on adult longhorn beetles Morimus funereus (Coleoptera, Cerambycidae). Based on a wavelet analysis of the local field potentials (LFPs), our study showed for the first time that the effects of prolonged and repeated exposure to the ELF MF on the LFPs were irreversible within investigated time frame. The relative wavelet energy (RWE) of 4-8 Hz frequency band was significantly increased after sine ELF MF (SnMF)/square ELF MF (SqMF) in comparison to the control value. The RWE of slower oscillations (1-2Hz) was significantly decreased after the repeated exposures to either SnMF or SqMF. The SqMF induced decreasing of the faster waves in the range of 64-128 Hz. However, we did not prove with presented methods that exposure to the SMF for 5 min produces any effects on the neuronal population activity. This study has proved the wavelet transform as a valuable tool for measuring the effects of SMF and ELF MF on the neuronal population activity in M. funereus antennal lobe.

Ouabain modulation of snail Br neuron bursting activity after the exposure to 10 mT static magnetic field revealed by Higuchi fractal dimension.

Aim of this study was to investigate the application of normalized mean of the empirical Higuchi fractal dimension (FD) distributions, as a new approach to analyze the spontaneous bioelectrical activity of garden snail (Helix pomatia) Br neuron. The effect of ouabain on modulation of Br neuron bursting activity before and after the exposure to 10 mT static magnetic field (SMF) was observed by analyzing the following parameters: action potential (AP), interspike interval (ISI) and interbursting interval (IBI) components. Normalized mean of the empirical FD distributions were formed for the following experimental conditions: Control 1, Ouabain 1, Control 2, SMF 2, ASMF 2, Control 3, SMF 3 and Ouabain ASMF 3. Our main results have shown that ouabain without SMF induced increase in participation of AP and a decrease in participation of IBI components compared to the first control condition. However, in the presence of 10 mT SMF, ouabain-induced changes of measured parameters of Br neuron activity were less pronounced compared to the third control condition. We have shown that normalized mean of the empirical FD distributions is a useful method for detecting the changes in AP, ISI, and IBI components of complex bursting activity in altered physiological states.

Non-toxic orange carbon dots stimulate photosynthesis and CO2 assimilation in hydroponically cultivated green beans (Phaseolus vulgaris).

Continuous increasing leaf photosynthesis may enhance plant yield. As an evolutionary property, plants use less photosynthetic capacity than is theoretically possible. Plant nanobionics is a bioengineering field that improves plant functions using nanoparticles. We applied orange carbon dots (o-CDs) onto the foliage of green beans (Phaseolus vulgaris ) grown in hydroponics to improve their photosynthetic performance and CO2 assimilation. Photosynthesis parameters, photosynthetic pigments content, total phenolic content (TPC) and antioxidative activity (TAA) were measured. Results show that photosynthetic pigments remained unchanged, while photosynthesis was improved. Both o-CDs concentrations decreased TPC and TAA. The light response curve showed higher CO2 assimilation at both o-CDs concentrations, particularly at lower light intensity. Correlation analysis confirmed increased CO2 binding and assimilation at 1mg L-1 . This study demonstrated the potential of using o-CDs as a safe biostimulator through photosynthesis increase and CO2 assimilation without toxic effects on plants. This may stimulate yield increase that paves the way for their agricultural application.

Auxin- and pH-induced guttation in Phycomyces sporangiophores: relation between guttation and diminished elongation growth.

Guttation, the formation of exudation water, is widespread among plants and fungi, yet the underlying mechanisms remain largely unknown. We describe the conditions for inducing guttation in sporangiophores of the mucoracean fungus, Phycomyces blakesleeanus. Cultivation on peptone-enriched potato dextrose agar elicits vigorous guttation mainly below the apical growing zone, while sporangiophores raised on a glucose-mineral medium manifest only moderate guttation. Mycelia do not guttate irrespective of the employed media. The topology of guttation droplets allows identifying the non-growing part of the sporangiophore as a guttation zone, which responds to humidity and medium composition in ways that become relevant for turgor homeostasis and thus the sensor physiology of the growing zone. Apparently, the entire sporangiophore, rather than exclusively the growing zone, participates in signal reception and integration to generate a common growth output. Exogenous auxin applied to the growing zones elicits two correlated responses: (i) formation of guttation droplets in the growing and transition zones below the sporangium and (ii) a diminution of the growth rate. In sporangiophore populations, guttation-induction by exogenous control buffer occurs at low frequencies; the bias for guttation increases with increasing auxin concentration. Synthetic auxins and the transport inhibitor NPA suppress guttation completely, but leave growth rates largely unaffected. Mutants C2 carA and C148 carA madC display higher sensitivities for auxin-induced guttation compared to wild type. A working model for guttation includes aquaporins and mechanosensitive ion channels that we identified in Phycomyces by sequence domain searches.

Cortico-pontine theta carrier frequency phase shift across sleep/wake states following monoaminergic lesion in rat.

This study was aimed to explore the sleep/wake states related cortico-pontine theta carrier frequency phase shift following a systemically induced chemical axotomy of the monoaminergic afferents within a brain of the freely moving rats. Our experiments were performed in 14 adult, male Sprague Dawley rats, chronically implanted for sleep recording. We recorded sleep during baseline condition, following sham injection (saline i.p. 1 ml/kg), and every week for 5 weeks following injection of the systemic neurotoxins (DSP-4 or PCA; 1 ml/kg, i.p.) for chemical axotomy of the locus coeruleus (LC) and dorsal raphe (DR) axon terminals. After sleep/wake states identification, FFT analysis was performed on 5 s epochs. Theta carrier frequency phase shift (∆Φ) was calculated for each epoch by averaging theta Fourier component phase shifts, and the ∆Φ values were plotted for each rat in control condition and 28 days following the monoaminergic lesions, as a time for permanently established DR or LC chemical axotomy. Calculated group averages have shown that ∆Φ increased between pons and cortex significantly in all sleep/wake states (Wake, NREM and REM) following the monoaminergic lesions, with respect to controls. Monoaminergic lesions established the pontine leading role in the brain theta oscillations during all sleep/wake states.

Signal transduction in Phycomyces sporangiophores: columella as a novel sensory organelle mediating auxin-modulated growth rate and membrane potential.

The growing zone (GZ) of the unicellular coenocytic sporangiophore of Phycomyces blakesleeanus represents the site of stimulus reception (light, gravity, gas) and stimulus response, i.e., local modulations of the elongation growth, which may result, in dependence of the stimulus direction, in tropic bending. Until now, evidence for a possible participation of the columella in sensory reception is absent. We confirm with light microscopy earlier studies that show that the GZ and the columella are not separated by a membrane or cell wall, but rather form a spatial continuum that allows free exchange of cytoplasm and organelle transport. Evidence is presented that the columella is responsive to external stimuli. Columellae, from which spores and sporangial cell wall had been removed, respond to exogenous auxin with a local depolarization of the membrane potential and an increased growth rate of the GZ. In contrast, auxin applied to the GZ causes a decrease of the growth rate irrespective of the presence or absence of sporangia. The response pattern is specific and relevant for the sensory reception of Phycomyces, because the light-insensitive mutant C148carAmadC, which lacks the RAS-GAP protein MADC, displays abnormal IAA sensitivity and membrane depolarization. We argue that the traditional concept of the GZ as the only stimulus-sensitive zone should be abandoned in favor of a model in which GZ and columella operate as a single entity capable to orchestrate a multitude of stimulus inputs, including auxin, to modulate the membrane potential and elongation growth of the GZ.

Surrogate data modeling the relationship between high frequency amplitudes and Higuchi fractal dimension of EEG signals in anesthetized rats.

We used spectral analysis and Higuchi fractal dimension (FD) to correlate the EEG spectral characteristics of the sensorimotor cortex, hippocampus, and pons with their corresponding EEG signal complexities in anesthetized rats. We have explored the quantitative relationship between the mean FDs and EEG wide range high frequency (8-50 Hz) activity during ketamine/xylazine versus nembutal anesthesia at surgical plane. Using FD we detected distinct inter-structure complexity pattern and uncovered for the first time that the polygraphically and behaviorally defined anesthetized state at surgical plane as equal during experiment in two anesthetic regimens, is not the same with respect to the degree of neuronal activity (degree of generalized neuronal inhibition achieved) at different brain levels. Using the correlation of certain brain structure EEG spectral characteristics with their corresponding FDs, and the surrogate data modeling, we determined what particular frequency band contributes to EEG complexities in ketamine/xylazine versus nembutal anesthesia. In this study we have shown that the quantitative relationship between higher frequency EEG amplitude and EEG complexity is the best-modeled by surrogate data as a 3rd order polynomial. On the base of our EEG amplitude/EEG complexity relationship model, and the evidenced spectral differences in ketamine versus nembutal anesthesia we have proved that higher amplitudes of sigma, beta, and gamma frequency in ketamine anesthesia yields to higher FDs.

Photosynthesis Enhancement in Maize via Nontoxic Orange Carbon Dots.

The sustained increase in leaf photosynthesis may increase crop yield. Due to many limitations, plants use much less photosynthetic capacity than is theoretically possible. Plant nanobionics investigates nanoparticle application in living plants, which improves certain plant functions. We synthesized and tested nontoxic orange carbon dots (o-CDs) for the photosynthetic efficiency increase in maize (Zea mays L.). We applied o-CDs foliarly or by adding to the growth solution. The photosynthetic parameters and content of photosynthetic pigments were recorded. The total phenolic content (TPC) and total antioxidant activity (TAA) were measured to monitor the plant antioxidant response to o-CDs. The photosynthetic parameters' values were higher for foliar than for solution application. The 1 mg/L o-CDs applied foliarly and 5 mg/L in solution increased photosynthetic parameters in leaves. The o-CDs raised photosynthetic pigments. The TAA and TPC results indicate reduced antioxidant activity in the plant organs more exposed to o-CDs, depending on the way of application.

Tangent algorithm for photogravitropic balance in plants and Phycomyces blakesleeanus: Roles for EHB1 and NPH3 of Arabidopsis thaliana.

Plant organs that are exposed to continuous unilateral light reach in the steady-state a photogravitropic bending angle that results from the mutual antagonism between the photo- and gravitropic responses. To characterize the interaction between the two tropisms and their quantitative relationship we irradiated seedlings of Arabidopsis thaliana that were inclined at various angles and determined the fluence rates of unilateral blue light required to compensate the gravitropism of the inclined hypocotyls. We found the compensating fluence rates to increase with the tangent of the inclination angles (0° < γ < 90° or max. 120°) and decrease with the cotangent (90°< γ < 180° or max. 120°of the inclination angles. The tangent dependence became also evident from analysis of previous data obtained with Avena sativa and the phycomycete fungus, Phycomyces blakesleeanus. By using loss-of function mutant lines of Arabidopsis, we identified EHB1 (enhanced bending 1) as an essential element for the generation of the tangent and cotangent relationships. Because EHB1 possesses a C2-domain with two putative calcium binding sites, we propose that the ubiquitous calcium dependence of gravi- and phototropism is in part mediated by Ca2+-bound EHB1. Based on a yeast-two-hybrid analysis we found evidence that EHB1 does physically interact with the ARF-GAP protein AGD12. Both proteins were reported to affect gravi- and phototropism antagonistically. We further showed that only AGD12, but not EHB1, interacts with its corresponding ARF-protein. Evidence is provided that AGD12 is able to form homodimers as well as heterodimers with EHB1. On the basis of these data we present a model for a mechanism of early tropism events, in which Ca2+-activated EHB1 emerges as the central processor-like element that links the gravi- and phototropic transduction chains and that generates in coordination with NPH3 and AGD12 the tangent / cotangent algorithm governing photogravitropic equilibrium.

Ecological risk assessment for different macrophytes and fish species in reservoirs using biota-sediment accumulation factors as a useful tool.

Metal content was evaluated in the sediment, macrophytes and fish in the Medjuvrsje reservoir (Western Serbia). Concentrations of 16 trace elements (Ag; Al; As; B; Ba; Cd; Co; Cr; Cu; Fe; Li; Mn; Ni; Pb; Sr; Zn) were analysed in the sediment, macrophytes and fish of an aquatic ecosystem. Five macrophyte species and three fish tissues (liver, muscle, gills) from five fish species (freshwater bream, common nase, Prussian carp, chub, wels catfish) were sampled and the metal content was analysed with ICP-OES. The sediment concentrations of Cu, Cd, and Zn exceeded the Canadian sediment quality guidelines while concentrations of Cr and Ni were above the Netherlands' target values. Bioaccumulation factors (BSAF) were calculated for analysed macrophytes and fish tissue. The BSAF had higher values for macrophytes for all investigated elements except for Cu and Zn; Cu had a higher value in the liver of the freshwater bream (0.823) and Zn had a higher value in the liver of freshwater bream (0.914) and chub (0.834) as well as in gills of Prussian carp (2.58) and chub (1.26). Potamogeton pectinatus, Ceratophylum demersum and the root of Phragmites communis showed higher accumulation of elements than Trapa natans and Potamogeton fluitans and the body of P. communis. The highest BSAF values for Ba, Mn, Sr and Ni were recorded in the gills. Cd and Cu had the highest BSAF values in the liver. Results confirmed that particular macrophyte and fish species could be a good indicator of reservoir water and sediment pollution.

The effect of auxin (indole-3-acetic acid) on the growth rate and tropism of the sporangiophore of Phycomyces blakesleeanus and identification of auxin-related genes.

The roles of fungal auxins in the regulation of elongation growth, photo-, and gravitropism are completely unknown. We analyzed the effects of exogenous IAA (indole-3-acetic acid), various synthetic auxins including 1-NAA (1-naphthaleneacetic acid) and 2,4-D (2,4-dichlorophenoxyacetic acid), and the auxin transport inhibitor NPA (N-1-naphtylphtalamic acid) on the growth rate and bending of the unicellular sporangiophore of the zygomycete fungus, Phycomyces blakesleeanus. Sporangiophores that were submerged in an aqueous buffer responded to IAA with a sustained enhancement of the growth rate, while 1-NAA, 2,4-D, and NPA elicited an inhibition. In contrast, sporangiophores kept in air responded to IAA with a 20 to 40% decrease of the growth rate, while 1-NAA and NPA elicited an enhancement. The unilateral and local application of IAA in the growing zone of the sporangiophore elicited in 30 min a moderate negative tropic bending in wild type C2 and mutant C148madC, which was, however, partially masked by a concomitant avoidance response caused by the aqueous buffer. Auxin transport-related genes ubiquitous in plants were found in a BLAST search of the Phycomyces genome. They included members of the AUX1 (auxin influx carrier protein 1), PILS (PIN-LIKES, auxin transport facilitator protein), and ABCB (plant ATP-binding cassette transporter B) families while members of the PIN family were absent. Our observations imply that IAA represents an intrinsic element of the sensory transduction of Phycomyces and that its mode of action must very likely differ in several respects from that operating in plants.

Application of Higuchi's fractal dimension from basic to clinical neurophysiology: A review.

BACKGROUND AND OBJECTIVE: For more than 20 years, Higuchi's fractal dimension (HFD), as a nonlinear method, has occupied an important place in the analysis of biological signals. The use of HFD has evolved from EEG and single neuron activity analysis to the most recent application in automated assessments of different clinical conditions. Our objective is to provide an updated review of the HFD method applied in basic and clinical neurophysiological research. METHODS: This article summarizes and critically reviews a broad literature and major findings concerning the applications of HFD for measuring the complexity of neuronal activity during different neurophysiological conditions. The source of information used in this review comes from the PubMed, Scopus, Google Scholar and IEEE Xplore Digital Library databases. RESULTS: The review process substantiated the significance, advantages and shortcomings of HFD application within all key areas of basic and clinical neurophysiology. Therefore, the paper discusses HFD application alone, combined with other linear or nonlinear measures, or as a part of automated methods for analyzing neurophysiological signals. CONCLUSIONS: The speed, accuracy and cost of applying the HFD method for research and medical diagnosis make it stand out from the widely used linear methods. However, only a combination of HFD with other nonlinear methods ensures reliable and accurate analysis of a wide range of neurophysiological signals.

Common barbel (Barbus barbus) as a bioindicator of surface river sediment pollution with Cu and Zn in three rivers of the Danube River Basin in Serbia.

River sediments are a major source of metal contamination in aquatic food webs. Due to the ability of metals to move up the food chain, fishes, occupying higher trophic levels, are considered to be good environmental indicators of metal pollution. The aim of this study was to analyze the metal content in tissues of the common barbel (Barbus barbus), a rheophilous cyprinid fish widely distributed in the Danube Basin, in order to find out if it can be used as a bioindicator of the metal content in the river sediment. We analyzed bioavailable concentrations of 15 elements (Al, As, B, Ba, Co, Cr, Cu, Hg, Mn, Mo, Ni, Pb, Se, Sr, and Zn) in sediments of the Danube (D), the Zapadna Morava (ZM), and the Juzna Morava (JM) using the inductively coupled plasma spectroscopy (ICP-OES). The barbel specimens were collected in the proximity of sediment sampling sites for the analysis of metals in four tissues, gills, muscle, intestine, and liver. The sediment analysis indicated that the ZM is the most polluted with Cu, Ni, and Zn compared to other two rivers. The JM had the lowest concentrations of almost all observed elements, while the Danube sediments were mainly characterized by higher concentrations of Pb. The fish from the ZM had the highest concentration of Cu and Ni in the liver and intestine, and of Zn in the muscle tissue, which was in accordance with the concentrations of these metals in the sediment. Scanning electron microscopy/energy dispersive X-ray spectroscopy (SEM-EDS) was used for further analyses of metal interactions with fish tissues. The results suggest that the barbel can potentially be used as a bioindicator of sediment quality with respect to metal contamination.

Estimation of parameter kmax in fractal analysis of rat brain activity.

We recorded electrocortical activity in anesthetized rats and constructed k(max) new self-similar time series, applying Higuchi's algorithm. The aim of this study was to estimate value of the parameter k(max) in order to obtain fractal dimension values as an optimum measure of biosignal change. After our analysis, electrocortical activity recordings resulted in a family of curves f(k(max)). Three regions could be distinguished 2 < or = k(max) < 8, with a U-shape; 8 < or = k(max) < or = 30, with a steeper quasilinear increase; and k(max) > or = 30, with a smaller slope quasilinear increase. We suggest the optimum region for k(max): 8 < k(max) < 18, specifically k(max) = 8.

Spectral analysis of cerebellar activity after acute brain injury in anesthetized rats.

We investigated cerebellar electrocortical activity before and after unilateral brain injury in anesthetized rats. Spectral analysis of cerebellar activity was obtained by Fast Fourier Transformation. There was a dominance of delta frequency range, while the wide gamma range presented no more than 5% of the total mean power spectra of cerebellar activity before brain injury. A few minutes after brain injury and within the first 90 minutes, there was a decrease of total mean power spectra and a relative decrease of delta range power to about 30%, some increase of beta range, and an increase of gamma range to 20-25%. Relative increase of gamma range in the cerebellar mean power spectra was still present 120 min after the brain injury, while other changes started to diminish. We suggest that spectral changes within slow and fast (gamma) frequency ranges of cerebellar activity may be indicators of the brain state after acute injury.

Detection of DNA mutations based on analysis of multiple wavelength excitation/emission fluorescence kinetics curves in real-time PCR.

The key method for therapies of various cancer types could be the molecular-targeted therapy, based on individual gene profile for each patient. One of the main procedures used for genetic testing is the real-time polymerase chain reaction (real-time PCR). Physical principle behind real-time PCR procedure is the fluorescence. Fluorescence labeled probes (primers) is attached to quenchers. Upon reaction of polymerization, quenchers are removed, and the fluorescence emission intensity increases in time. Emission spectra shape and its maximum position can differ if the fluorophore was present in different microenvironment. That property is widely exploited in fluorescence spectroscopy and chromatography. This paper, for the first time, describes utilization of full spectroscopic potential of multichannel excitation/emission filter sets in real-time PCR device. Instead of monitoring fluorescence intensity in time for a single fluorescence emission channel, the ratio values of three different kinetics curves were calculated and analyzed by applying k-means clustering and dendrogram analysis. Obtained results have shown that described analytical improvement provides identification of nine different groups of mutations if the commercial QIAGEN® EGFR PCR Kit was used. Method can be applied to any kit, capable to simultaneously detect several different mutations.

Spectral and fractal analysis of cerebellar activity after single and repeated brain injury.

The cerebellum, even when not directly damaged, is potentially interesting for understanding the adaptive responses to brain injury. Cerebellar electrocortical activity (ECoG) in rats was studied using spectral and fractal analysis after single and repeated unilateral injury of the parietal cortex. Local field potentials of cerebellar paravermal cortex were recorded before brain injury, in the acute phase (up to 2.5 hours) after a first injury of anesthetized rats, and then before and after second, third, and, in some cases, fourth injury. Relative gamma power (32.1-128.0 Hz) and fractal dimension of ECoGs were temporarily increased after the first injury. However, there was a permanent mild increase in gamma activity and a mild increase in the fractal dimension of cerebellar activity as a chronic change after repeated remote brain injury. There was a negative linear correlation between the normalized difference in fractal dimensions and normalized difference in gamma powers of cerebellar activity only in the case of repeated brain injury. This is the first study showing that correlation between the parameters of spectral and fractal analyses of cerebellar activity can discriminate between single and repeated brain injuries, and is, therefore, a promising approach for identifying specific pathophysiological states.