Stimulation of antioxidant activity and γ-aminobutyric acid synthesis in germinated wheat grain Triticum aestivum L. by ultrasound: Increasing the nutritional value of the product.

The use of ultrasound to intensify the germination process of Triticum aestivum L. wheat was studied. This method of controlled germination can be used in several sectors of food industry, in particular in bakery. The effect of low-frequency ultrasound (20 kHz) at different intensities and duration on the germination process of Triticum aestivum L. wheat was systematically studied. We have found that 3-minute processing at 227 W/l output reduces the duration of wheat grain germination by 25% (12 ± 2 h) compared to the control samples. The use of ultrasound stimulated γ-aminobutyric acid (GABA) synthesis (18.9 ± 0.5 mg/100 g), increased the antioxidant activity (AOA) (2.86 ± 0.2 mg/g Trolox equivalents) and the amount of flavonoids (0.19 ± 0.03 mg QE/g). The SEM analysis of powder particles of whole-wheat flour made from wheat germinated with ultrasound exposure showed densely packed aggregates of protein matrix. To sum up, controlled ultrasound during wheat grain germination increases the amount of GABA and AOA. The whole-wheat flour is useful for food enrichment.

Bulky Adducts in Clustered DNA Lesions: Causes of Resistance to the NER System.

The nucleotide excision repair (NER) system removes a wide range of bulky DNA lesions that cause significant distortions of the regular double helix structure. These lesions, mainly bulky covalent DNA adducts, are induced by ultraviolet and ionizing radiation or the interaction between exogenous/endogenous chemically active substances and nitrogenous DNA bases. As the number of DNA lesions increases, e.g., due to intensive chemotherapy and combination therapy of various diseases or DNA repair impairment, clustered lesions containing bulky adducts may occur. Clustered lesions are two or more lesions located within one or two turns of the DNA helix. Despite the fact that repair of single DNA lesions by the NER system in eukaryotic cells has been studied quite thoroughly, the repair mechanism of these lesions in clusters remains obscure. Identification of the structural features of the DNA regions containing irreparable clustered lesions is of considerable interest, in particular due to a relationship between the efficiency of some antitumor drugs and the activity of cellular repair systems. In this review, we analyzed data on the induction of clustered lesions containing bulky adducts, the potential biological significance of these lesions, and methods for quantification of DNA lesions and considered the causes for the inhibition of NER-catalyzed excision of clustered bulky lesions.

Recognition and removal of clustered DNA lesions via nucleotide excision repair.

Clustered damage of DNA consists of two or more lesions located within one or two turns of the DNA helix. Clusters consisting of lesions of various structures can arise under the influence of strong damaging factors, especially if the cells have a compromised repair status. In this work, we analyzed how the presence of an analog of the apurinic/apyrimidinic site - a non-nucleoside residue consisting of diethylene glycol phosphodiester (DEG) - affects the recognition and removal of a bulky lesion (a non-nucleoside site of the modified DNA strand containing a fluorescein residue, nFlu) from DNA by a mammalian nucleotide excision repair system. Here we demonstrated that the efficiency of nFlu removal decreases in the presence of DEG in the complementary strand and is completely suppressed when the DEG is located opposite the nFlu. By contrast, protein factor XPC-RAD23B, which initiates global genomic nucleotide excision repair, has higher affinity for DNA containing clustered damage as compared to DNA containing a single bulky lesion; the affinity of XPC strengthens as the positions of DEG and nFlu become closer. The changes in the double-stranded DNA's geometry caused by the presence of clustered damage were also assessed. The obtained experimental data together with the results of molecular dynamics simulations make it possible to get insight into the structural features of DNA containing clustered lesions that determine the efficiency of repair. Speaking more broadly, this study should help to understand the probable fate of bulky adduct-containing clusters of various topologies in the mammalian cell.

Spatially resolved spectroscopic ion temperature measurements at plasma edge of the T-10 tokamak.

Spectroscopic diagnostics of the edge ion temperature were developed on the T-10 tokamak. Spatially resolved measurements of C5+ and other ionization states of carbon spectral line shapes are provided. Spectra were measured with high spectral resolution using 14 lines of sight in one poloidal section of the tokamak. Each measured line-integrated spectrum contains a combination of multiple local spectra with corresponding values of ion temperature. Modeling of spatial distribution of line emissivity and spectral line shapes along the lines of sight allows the reconstruction of the ion temperature profile on the basis of the closest match of measured and modeled spectra. The fine structure of spectral line, Zeeman effect, and apparatus function are taken into account during data processing. Obtained ion temperature profiles, Ti(r), at the plasma edge are in good agreement with ion temperature profiles measured by Charge eXchange Recombination Spectroscopy (CXRS) diagnostics of T-10. Use of the CXRS equipment for measurements of passive spectra can provide additional information on the temporal evolution of the edge ion temperature. Developed diagnostics provide necessary data for the research of geodesic acoustic modes, which are strongly dependent on plasma edge ion temperature.

Charge exchange recombination spectroscopy on the T-10 tokamak.

The charge exchange recombination spectroscopy (CXRS) diagnostics on the T-10 tokamak is described. The system is based on a diagnostic neutral beam and includes three high etendue spectrometers designed for the ITER edge CXRS system. A combined two-channel spectrometer is developed for simultaneous measurements of two beam-induced spectral lines using the same lines of sight. A basic element of the combined spectrometer is a transmitting holographic grating designed for the narrow spectral region 5291 ± 100 Å. The whole CXRS system provides simultaneous measurements of two CXRS impurity spectra and Hα beam line. Ion temperature measurements are routinely provided using the C(6+) CXRS spectral line 5291 Å. Simultaneous measurements of carbon densities and one more impurity (oxygen, helium, lithium etc.) are carried out. Two light collecting systems with 9 lines of sight in each system are used in the diagnostics. Spatial resolution is up to 2.5 cm and temporal resolution of 1 ms is defined by the diagnostic neutral beam diameter and pulse duration, respectively. Experimental results are shown to demonstrate a wide range of the CXRS diagnostic capabilities on T-10 for investigation of impurity transport processes in tokamak plasma. Developed diagnostics provides necessary experimental data for studying of plasma electric fields, heat and particle transport processes, and for investigation of geodesic acoustic modes.

Ca(2+) -activated K(+) current is essential for maintaining excitability and gene transcription in early embryonic cardiomyocytes.

AIM: Activity of early embryonic cardiomyocytes relies on spontaneous Ca(2+) oscillations that are induced by interplay between sarcoplasmic reticulum (SR) - Ca(2+) release and ion currents of the plasma membrane. In a variety of cell types, Ca(2+) -activated K(+) current (IK(Ca) ) serves as a link between Ca(2+) signals and membrane voltage. This study aimed to determine the role of IK (Ca) in developing cardiomyocytes. METHODS: Ion currents and membrane voltage of embryonic (E9-11) mouse cardiomyocytes were measured by patch clamp; [Ca(2+) ]i signals by confocal microscopy. Transcription of specific genes was measured with RT-qPCR and Ca(2+) -dependent transcriptional activity using NFAT-luciferase assay. Myocyte structure was assessed with antibody labelling and confocal microscopy. RESULTS: E9-11 cardiomyocytes express small conductance (SK) channel subunits SK2 and SK3 and have a functional apamin-sensitive K(+) current, which is also sensitive to changes in cytosolic [Ca(2+) ]i . In spontaneously active cardiomyocytes, inhibition of IK (Ca) changed action and resting potentials, reduced SR Ca(2+) load and suppressed the amplitude and the frequency of spontaneously evoked Ca(2+) oscillations. Apamin caused dose-dependent suppression of NFAT-luciferase reporter activity, induced downregulation of a pattern of genes vital for cardiomyocyte development and triggered changes in the myocyte morphology. CONCLUSION: The results show that apamin-sensitive IK (Ca) is required for maintaining excitability and activity of the developing cardiomyocytes as well as having a fundamental role in promoting Ca(2+) - dependent gene expression.

Hunting for origins of migraine pain: cluster analysis of spontaneous and capsaicin-induced firing in meningeal trigeminal nerve fibers.

Trigeminal nerves in meninges are implicated in generation of nociceptive firing underlying migraine pain. However, the neurochemical mechanisms of nociceptive firing in meningeal trigeminal nerves are little understood. In this study, using suction electrode recordings from peripheral branches of the trigeminal nerve in isolated rat meninges, we analyzed spontaneous and capsaicin-induced orthodromic spiking activity. In control, biphasic single spikes with variable amplitude and shapes were observed. Application of the transient receptor potential vanilloid 1 (TRPV1) agonist capsaicin to meninges dramatically increased firing whereas the amplitudes and shapes of spikes remained essentially unchanged. This effect was antagonized by the specific TRPV1 antagonist capsazepine. Using the clustering approach, several groups of uniform spikes (clusters) were identified. The clustering approach combined with capsaicin application allowed us to detect and to distinguish "responder" (65%) from "non-responder" clusters (35%). Notably, responders fired spikes at frequencies exceeding 10 Hz, high enough to provide postsynaptic temporal summation of excitation at brainstem and spinal cord level. Almost all spikes were suppressed by tetrodotoxin (TTX) suggesting an involvement of the TTX-sensitive sodium channels in nociceptive signaling at the peripheral branches of trigeminal neurons. Our analysis also identified transient (desensitizing) and long-lasting (slowly desensitizing) responses to the continuous application of capsaicin. Thus, the persistent activation of nociceptors in capsaicin-sensitive nerve fibers shown here may be involved in trigeminal pain signaling and plasticity along with the release of migraine-related neuropeptides from TRPV1 positive neurons. Furthermore, cluster analysis could be widely used to characterize the temporal and neurochemical profiles of other pain transducers likely implicated in migraine.

Cortical spreading depression induces oxidative stress in the trigeminal nociceptive system.

Indirect evidence suggests the increased production of reactive oxygen species (ROS) in migraine pathophysiology. In the current study we measured lipid peroxidation product in the rat cortex, trigeminal ganglia and meninges after the induction of cortical spreading depression (CSD), a phenomenon known to be associated with migraine aura, and tested nociceptive firing triggered by ROS in trigeminal nerves ex vivo. Application of KCl to dura mater in anesthetized rats induced several waves of CSD recorded by an extracellular electrode in the cortex. Following CSD, samples of cortex (affected regions were identified with blood oxygen level-dependent (BOLD) functional magnetic resonance imaging (fMRI)), meninges from left and right hemispheres and trigeminal ganglia were taken for biochemical analysis. We found that CSD increased the level of the lipid peroxidation product malondialdehyde (MDA) in the ipsilateral cerebral cortex and meninges, but also in both ipsi- and contralateral trigeminal ganglia. In order to test the pro-nociceptive action of ROS, we applied the mild oxidant hydrogen peroxide to isolated rat hemiskull preparations including preserved trigeminal innervations. Application of hydrogen peroxide to meninges transiently enhanced electrical spiking activity of trigeminal nerves showing a pro-nociceptive action of ROS. In the presence of hydrogen peroxide trigeminal nerves still responded to capsaicin by burst of spiking activity indicating integrity of neuronal structures. The action of hydrogen peroxide was mediated by TRPA1 receptors as it was abolished by the specific TRPA1 antagonist TCS-5861528. Using dorsal root ganglion sensory neurons as test system we found that hydrogen peroxide promoted the release of the migraine mediator calcitonin gene-related peptide (CGRP), which we previously identified as a trigger of delayed sensitization of trigeminal neurons. Our data suggest that, after CSD, oxidative stress spreads downstream within the trigeminal nociceptive system and could be involved in the coupling of CSD with the activation of trigeminovascular system in migraine pathology.

Advanced Thomson scattering system for high-flux linear plasma generator.

An advanced Thomson scattering system has been built for a linear plasma generator for plasma surface interaction studies. The Thomson scattering system is based on a Nd:YAG laser operating at the second harmonic and a detection branch featuring a high etendue (f/3) transmission grating spectrometer equipped with an intensified charged coupled device camera. The system is able to measure electron density (n(e)) and temperature (T(e)) profiles close to the output of the plasma source and, at a distance of 1.25 m, just in front of a target. The detection system enables to measure 50 spatial channels of about 2 mm each, along a laser chord of 95 mm. By summing a total of 30 laser pulses (0.6 J, 10 Hz), an observational error of 3% in n(e) and 6% in T(e) (at n(e) = 9.4 × 10(18) m(-3)) can be obtained. Single pulse Thomson scattering measurements can be performed with the same accuracy for n(e) > 2.8 × 10(20) m(-3). The minimum measurable density and temperature are n(e) < 1 × 10(17) m(-3) and T(e) < 0.07 eV, respectively. In addition, using the Rayleigh peak, superimposed on the Thomson scattered spectrum, the neutral density (n(0)) of the plasma can be measured with an accuracy of 25% (at n(0) = 1 × 10(20) m(-3)). In this report, the performance of the Thomson scattering system will be shown along with unprecedented accurate Thomson-Rayleigh scattering measurements on a low-temperature argon plasma expansion into a low-pressure background.

Increased non-quantal release of acetylcholine after inhibition of endocytosis by methyl-ß-cyclodextrin: the role of vesicular acetylcholine transporter.

We investigated the role of the vesicular acetylcholine transporter in the mechanism of non-quantal (non-vesicular) secretion of neurotransmitter in the neuromuscular synapse of the rat diaphragm muscle. Non-quantal secretion was estimated electrophysiologically by the amplitude of end-plate hyperpolarization after inhibition of cholinesterase and nicotinic receptors (H-effect) or measured by the optical detection of acetylcholine in the bathing solution. It was shown that 1 mM methyl-ß-cyclodextrin (MCD) reduced both endocytosis and, to much lesser extent, exocytosis of synaptic vesicles (SV) thereby increasing non-quantal secretion of acetylcholine with a concurrent decrease in axoplasm pH. During high-frequency stimulation of the motor nerve, that substantially increases vesicles exocytosis, the non-quantal secretion was further enhanced if the endocytosis of SV was blocked by MCD. In contrast, non-quantal secretion of acetylcholine did not increase when the MCD-treated neuromuscular preparations were superfused with either vesamicol, an inhibitor of vesicular transporter of acetylcholine, or sodium propionate, which decreases intracellular pH. These results suggest that the proton-dependent, vesamicol-sensitive vesicular transporters of acetylcholine, which become inserted into the presynaptic membrane during SV exocytosis and removed during endocytotic recycling of SV, play the major role in the process of non-quantal secretion of neurotransmitter.

Interaction of glutamate- and adenosine-induced decrease of acetylcholine quantal release at frog neuromuscular junction.

In a frog neuromuscular preparation of m. sartorius, glutamate had a reversible dose-dependent inhibitory effect on both spontaneous miniature endplate potentials (MEPP) and nerve stimulation-evoked endplate potentials (EPP). The effect of glutamate on MEPP and EPP is caused by the activation of metabotropic glutamate receptors, as it was eliminated by MCPG, an inhibitor of group I metabotropic glutamate receptors. The depression of evoked EPP, but not MEPP frequency was removed by inhibiting the NO production in the muscle by L-NAME and by ODQ that inhibits the soluble NO-sensitive guanylyl cyclase. The glutamate-induced depression of the frequency of spontaneous MEPP is apparently not caused by the stimulation of the NO cascade. The particular glutamate-stimulated NO cascade affecting the evoked EPP can be down-regulated also by adenosine receptors, as the glutamate and adenosine actions are not additive and application of adenosine partially prevents the further decrease of quantal content by glutamate. On the other hand, there is no obvious interaction between the glutamate-mediated inhibition of EPP and inhibitory pathways triggered by carbacholine and ATP. The effect of glutamate on the evoked EPP release might be due to NO-mediated modulation (phosphorylation) of the voltage-dependent Ca2+ channels at the presynaptic release zone that are necessary for evoked quantal release and open during EPP production.

Adenosine triphosphoric acid as a factor of nervous regulation of Na+/K+/2Cl- cotransport in rat skeletal muscle fibers.

Exogenous adenosine triphosphoric acid produces a biphasic effect on the resting membrane potential of muscle fibers in rat diaphragm. Depolarization of the sarcolemma observed 10 min after application of adenosine triphosphoric acid results from activation of Na(+)/K(+)/2Cl(-) cotransport. The increase in chloride cotransport is related to activation of postsynaptic P2Y receptors and protein kinase C. Repolarization of the membrane develops 40 min after treatment with adenosine triphosphoric acid and after 50 min the resting membrane potential almost returns the control level. This increase in the resting membrane potential of the sarcolemma is probably associated with activation of the Na(+)/K(+) pump and increase in membrane permeability for chlorine ions in response to long-term activity of Cl(-) cotransport. Thus, adenosine triphosphoric acid co-secreted with acetylcholine in the neuromuscular synapse probably plays a role in the regulation resting membrane potential and cell volume of muscle fibers.

Effect of dipeptide N-acetylaspartylglutamate on denervation-induced changes in the volume of rat skeletal muscle fibers.

N-acetylaspartylglutamate prevents the denervation-induced increase in the volume of muscle fibers in rat diaphragm, the phenomenon being more pronounced for the hydrolysable isomer. The effect of dipeptide manifested against the background of blockade of metabotropic glutamate receptors. It was hypothesized that N-acetylaspartylglutamate is involved in the regulation of the volume of skeletal muscle fibers via activation of ionotropic receptors by both dipeptide and glutamate molecules.

Effect of oxotremorine on resting membrane potential and cell volume in skeletal muscle fibers in rats after in vivo blockade of NO-synthase.

Denervation of rat phrenic muscle or block of NO-synthase in vivo increased the cross-section area of muscle fibers and decreased membrane resting potential. Oxotremorine prevented the development of denervation-induced or denervation-like (i.e. induced by NO-synthase blockade) membrane depolarization and increase of the cross-sectional area of muscle fibers. Pirenzepine abolished the effects of oxotremorine. It was concluded that non-quantal acetylcholine can be involved in the regulation of skeletal muscle fiber volume via activation of M1 muscarinic receptors followed by NO synthesis.

[Pharmacological parameters of muscarinic cholinoreceptors in skeletal muscles]. / Farmakologicheskie osobennosti muskarinovykh cholinoretseptorov skeletnykh myshts.

It has been shown that bath application of muscarine delayed the early post-denervation depolarization in the muscle fibers incubated for 3 h in culture medium. The greatest reduction of the post-devervation depolarization was observed with 50 nmol/l muscarine. Atropine, a muscarinic antagonist, clozapine, a specific inhibitor of M1/M5-cholinergic receptors, and nitrocaramiphen, a M1-antagonist, completely removed the hyperpolarizing effect of muscarine. 4-DAMP, a specific inhibitor of M3-cholinergic receptors, himbacine, an antagonist of M2-cholinergic receptors, and tropicamide, a specific inhibitor of M2/M4-cholinergic receptors, failed to prevent the effect of muscarine. A M1/M2 muscarine agonists propargyl and but-2-ynyl esters of arecaidine had apparent muscarine-like effect. Nitrocaramiphen, and not himbacine, prevented the hyperpolarizing effect of these cholinomimetics. It is concluded that muscarine and esters of arecaidine delay the development of early postdenervation depolarization in M1-cholinergic receptors of skeletal muscle.

[Effect of glutamate on membrane potential and volume of the skeletal muscle fibers in rats following NO-synthase inhibition in vivo]. / Vliianie glutamata na membrannyi potentsial i ob''em skeletnykh myshechnykh volokon krysy posle blokady NO-sintazy in vivo.

Cross-sectional area (CSA) of muscle fibers incubated in culture medium 199 for 3 hours dramatically increases, whereas resting membrane potential (RMP) decreases compared to "freshly-isolated" muscles. Both glutamate and sodium nitroprusside prevent these changes. MK-801, a specific inhibitor of NMDA-receptors, eliminates protective effects of glutamate on both CSA and RMP. NO-synthase inhibition in vivo promotes an increase of initial CSA and decrease of mean RMP. Under these conditions, effects of glutamate and sodium nitroprusside on CSA and RMP of denervated muscles are less obvious. It has been concluded that synaptic glutamate is able to participate in regulation of RMP and cell volume in muscle fibers through the activation of postsynaptic NMDA-receptors and muscle NO-synthase.

Optimal cuts of langasite, La3Ga5SiO14 for SAW devices.

The results of a theoretical and experimental investigation of the SAW propagation characteristics in an optimal region of langasite defined by the Euler angles phi from -15 degrees to +10 degrees, theta from 120 degrees to 165 degrees, and psi from 20 degrees to 45 degrees are presented. Based on temperature coefficients of the elastic constants derived from experimental data, some optimal orientations of langasite characterized by high electromechanical coupling factor (k2), zero power flow angle (PFA) and low or zero temperature coefficient of frequency (TCF) were found. The SAW velocity in the region of interest is highly anisotropic; this results in a significant amount of diffraction, which must be taken into account in the search for orientations useful for SAW devices. An orientation having simultaneously zero PFA, zero TCF, negligible diffraction, and relatively high piezoelectric coupling has been found and verified experimentally. The experimental results are in excellent agreement with the calculated SAW characteristics. The frequency response of a SAW device fabricated on the optimal cut of langasite is presented and demonstrates that high performance SAW filters can be realized on this optimal cut of langasite.

Carbachol and acetylcholine delay the early postdenervation depolarization of muscle fibres through M1-cholinergic receptors.

The resting membrane potential (RMP) of denervated muscle fibres of rat diaphragm muscle is depolarized by approximately 8-10 mV during the first 3 h after nerve section and this early postdenervation depolarization is reduced substantially by the presence of 5x10(-8) M acetylcholine (ACh) or carbachol (CB). The muscarinic antagonist atropine (Atr; 5x10(-9) to 5x10(-6) M) reduced the effect of CB in a dose-dependent manner (K(i)=7x10(-8) M) and increased the rate of the early postdenervation depolarization. In lower doses (5x10(-7) M), Atr acted only in the presence of an allosteric stabilizator hexamethylene-bis-[dimethyl-(3-phtalimidopropyl)ammonium] (W-84). Also pirenzepine, a specific inhibitor of the M1 subtype of muscarinic receptor, blocked the action of CB in a dose-dependent manner with an apparent inhibition constant K(i)=1x10(-7) microM. DAMP, a specific M3 antagonist, was without effect on the muscle hyperpolarization induced by CB. CB also hyperpolarized the membrane potentials of muscles which were denervated for 1-3 days. It is concluded that ACh and CB protect the muscle fibres from early depolarization through M1-cholinergic receptors on the muscle membrane. These particular receptors can apparently mediate the 'trophic', non-impulse regulation of RMP in skeletal muscles when they are activated by acetylcholine released non-quantally.

The experimental and theoretical characterization of the SAW propagation properties for zinc oxide films on silicon carbide.

The surface acoustic wave (SAW) propagation properties of zinc oxide (ZnO) films on silicon carbide (SiC) have been theoretically and experimentally characterized in the film thickness-to-acoustic wavelength ratio range up to 0.12. The experimental characterization of the SAW propagation properties was performed with a linear array of interdigital transducer (IDT) structures. The measurements characterized the velocity and propagation loss of two surface modes, a generalized SAW (GSAW) mode with velocities between 6000 and 7000 m/s, and a high velocity Pseudo-SAW (HVPSAW) mode with velocities between 8500 and 12 500 m/s. The experimentally determined characteristics of the two waves have been compared with the results of calculations based on published data for SiC and ZnO. Simulation of wave characteristics was performed with various values of the elastic constant C(13), which is absent in the published set of material constants for SiC, within the interval permitted by the requirement of positive elastic energy in a hexagonal crystal. The best agreement between the measured and calculated propagation losses of the HVPSAW has been obtained for C(13) near zero. Although for the GSAW mode the calculated velocity dispersion has been found nearly insensitive to the value of C (13) and consistent with the experimental data, for the HVPSAW, some disagreement between measured and calculated velocities, which increased with ZnO film thickness, has been observed for any C(13 ) value. Theoretical analysis of HVPSAW has revealed the existence of a previously unknown high velocity SAW (HVSAW). The displacement components of this wave have been analyzed as functions of depth and confirmed its pure surface, one-partial character.