Temperature responses of dark respiration in relation to leaf sugar concentration

Changes in leaf sugar concentrations are a possible mechanism of short‐term adaptation to temperature changes, with natural fluctuations in sugar concentrations in the field expected to modify the heat sensitivity of respiration. We studied temperature‐response curves of leaf dark respiration in the temperate tree Populus tremula (L.) in relation to leaf sugar concentration (1) under natural conditions or (2) leaves with artificially enhanced sugar concentration. Temperature‐response curves were obtained by increasing the leaf temperature at a rate of 1°C min^?1. We demonstrate that respiration, similarly to chlorophyll fluorescence, has a break‐point at high temperature, where respiration starts to increase with a faster rate. The average break‐point temperature (T_RD) was 48.6 ± 0.7°C at natural sugar concentration. Pulse‐chase experiments with ¹⁴CO₂ demonstrated that substrates of respiration were derived mainly from the products of starch degradation. Starch degradation exhibited a similar temperature‐response curve as respiration with a break‐point at high temperatures. Acceleration of starch breakdown may be one of the reasons for the observed high‐temperature rise in respiration. We also demonstrate that enhanced leaf sugar concentrations or enhanced osmotic potential may protect leaf cells from heat stress, i.e. higher sugar concentrations significantly modify the temperature‐response curve of respiration, abolishing the fast increase of respiration. Sugars or enhanced osmotic potential may non‐specifically protect respiratory membranes or may block the high‐temperature increase in starch degradation and consumption in respiratory processes, thus eliminating the break‐points in temperature curves of respiration in sugar‐fed leaves.     

Influence of mitochondrial genome rearrangement on cucumber leaf carbon and nitrogen metabolism

The MSC16 cucumber (Cucumis sativus L.) mitochondrial mutant was used to study the effect of mitochondrial dysfunction and disturbed subcellular redox state on leaf day/night carbon and nitrogen metabolism. We have shown that the mitochondrial dysfunction in MSC16 plants had no effect on photosynthetic CO₂ assimilation, but the concentration of soluble carbohydrates and starch was higher in leaves of MSC16 plants. Impaired mitochondrial respiratory chain activity was associated with the perturbation of mitochondrial TCA cycle manifested, e.g., by lowered decarboxylation rate. Mitochondrial dysfunction in MSC16 plants had different influence on leaf cell metabolism under dark or light conditions. In the dark, when the main mitochondrial function is the energy production, the altered activity of TCA cycle in mutated plants was connected with the accumulation of pyruvate and TCA cycle intermediates (citrate and 2-OG). In the light, when TCA activity is needed for synthesis of carbon skeletons required as the acceptors for NH₄ ⁺ assimilation, the concentration of pyruvate and TCA intermediates was tightly coupled with nitrate metabolism. Enhanced incorporation of ammonium group into amino acids structures in mutated plants has resulted in decreased concentration of organic acids and accumulation of Glu.     

Effect of noise in processing of visual information

Background

Information transmission and processing in the nervous system has stochastic nature. Multiple factors contribute to neuronal trial-to-trial variability. Noise and variations are introduced by the processes at the molecular and cellular level (thermal noise, channel current noise, membrane potential variations, biochemical and diffusion noise at synapses etc). The stochastic processes are affected by different physical (temperature, electromagnetic field) and chemical (drugs) factors. The aim of this study was experimental investigation of hypotheses that increase in the noise level in the brain affects processing of visual information. Change in the noise level was introduced by an external factor producing excess noise in the brain.

Methods

An exposure to 450 MHz low-frequency modulated microwave radiation was applied to generate excess noise. Such exposure has been shown to increase diffusion, alter membrane resting potential, gating variables and intracellular Calcium efflux. Nine healthy volunteers passed the experimental protocol at the lower (without microwave) and the higher (with microwave) noise level. Two photos (visual stimuli) of unfamiliar, young male faces were presented to the subjects, one picture after another. The task was to identify later the photos from a group of six photos and to decide in which order they were presented. Each subject had a total of eight sessions at the lower and eight at the higher noise level. Each session consisted of 50 trials; altogether a subject made 800 trials, 400 at the lower and 400 at the higher noise level. Student t-test was applied for statistical evaluation of the results.

Results

Correct recognition of both stimuli in the right order was better at the lower noise level. All the subjects under investigation showed higher numbers of right answers in trials at the lower noise level. Average number of correct answers from n=400 trials with microwave exposure was 50.3, without exposure 54.4, difference 7.5%, p<0.002. No difference between results at the lower and the higher noise level was revealed in the case of only partly correct or incorrect answers.

Conclusions

Our experimental results showed that introduced excess noise reduced significantly ability of the nervous system in correct processing of visual information.

     

Quantitative analysis of photosynthetic carbon metabolism in protoplasts and intact leaves of barley. Determination of carbon fluxes and pool sizes of metabolites in different cellular compartments

Rates of carbon fluxes and pool sizes of photosynthetic metabolites in different cellular compartments of barley protoplasts were calculated from the time curves of their labeling in the medium of ¹⁴CO₂. Using membrane filtration procedure, kinetics of ¹⁴C incorporation into the products of steady-state photosynthesis was determined separately in chloroplasts, mitochondria and cytosol of barley protoplasts illuminated for different periods in the air containing ¹⁴CO₂. To extract the quantitative information, analytical labeling functions P(t) describing the dependence of ¹⁴C content in the primary, intermediate and end products of a linear reaction chain upon the duration of tracer feeding have been derived. The parameters of these functions represent pool sizes of metabolites and rates of carbon fluxes. The values of these parameters were determined by fitting the experimental labeling curves to the functions P(t) by means of non-linear regression procedure. To elucidate the possible effects of fractionation on the photosynthetic carbon metabolism, the parameters of protoplasts were compared with corresponding values in intact leaves of barley.     

Effect of low frequency modulated microwave exposure on human EEG: individual sensitivity

The aim of this study was to evaluate the effect of modulated microwave exposure on human EEG of individual subjects. The experiments were carried out on four different groups of healthy volunteers. The 450 MHz microwave radiation modulated at 7 Hz (first group, 19 subjects), 14 and 21 Hz (second group, 13 subjects), 40 and 70 Hz (third group, 15 subjects), 217 and 1000 Hz (fourth group, 19 subjects) frequencies was applied. The field power density at the scalp was 0.16 mW/cm(2). The calculated spatial peak SAR averaged over 1 g was 0.303 W/kg. Ten cycles of the exposure (1 min off and 1 min on) at fixed modulation frequencies were applied. All subjects completed the experimental protocols with exposure and sham. The exposed and sham-exposed subjects were randomly assigned. A computer also randomly assigned the succession of modulation frequencies. Our results showed that microwave exposure increased the EEG energy. Relative changes in the EEG beta1 power in P3-P4 channels were selected for evaluation of individual sensitivity. The rate of subjects significantly affected is similar in all groups except for the 1000 Hz group: in first group 3 subjects (16%) at 7 Hz modulation; in second group 4 subjects (31%) at 14 Hz modulation and 3 subjects (23%) at 21 Hz modulation; in third group 3 subjects (20%) at 40 Hz and 2 subjects (13%) at 70 Hz modulation; in fourth group 3 subjects (16%) at 217 Hz and 0 subjects at 1000 Hz modulation frequency.     

Effect of microwave radiation on human EEG at two different levels of exposure

This study is aimed at evaluating the effect of microwave radiation on human brain bioelectric activity at different levels of exposure. For this purpose, 450 MHz microwave exposure modulated at 40 Hz frequency was applied to a group of 15 healthy volunteers at two different specific absorption rate (SAR) levels: a higher level of 0.303 W/kg (field strength 24.5 V/m) and a lower level of 0.003 W/kg (field strength 2.45 V/m). Ten exposure cycles (1 min off and 1 min on) at fixed SAR values were applied. A resting eyes‐closed electroencephalogram (EEG) was continuously recorded. Results showed a statistically significant increase in the EEG power in the EEG beta2 (157%), beta1 (61%) and alpha (68%) frequency bands at the higher SAR level, and in the beta2 (39%) frequency band at the lower SAR level. Statistically significant changes were detected for six individual subjects in the EEG alpha band and four subjects in the beta1 and beta2 bands at the higher SAR level; three subjects were affected in the alpha, beta1 and beta2 bands at the lower SAR level. The study showed that decreasing the SAR 100 times reduced the related changes in the EEG three to six times and the number of affected subjects, but did not exclude the effect. Bioelectromagnetics 34:264–274, 2013. © 2012 Wiley Periodicals, Inc.     

Study of effects of low level microwave field by method of face masking

The aim of this study was to examine experimentally effects of low level, modulated microwaves on human central nervous system function utilizing the phenomenon of visual masking. Ten healthy volunteers, four males and six females, were exposed to electromagnetic field (450 MHz, 0.16 mW/cm2) with 7 Hz modulation frequency. Two photo series (visual stimuli) of unfamiliar, young male faces were presented to the subjects, one picture after another. All the photos were frontal views of unfamiliar faces, which could be recognized only by their unique combinations of features. The task was to identify the pictures from a group of six photos and to decide which order they were presented in. The phenomenon of visual masking is revealed as anamorphosis in subject's perception of two instantaneous visual stimuli presented within a short time interval. When both stimuli were to be recognized correctly and put in the right order, there was a statistically significant difference (P < 0.05) between the identification of the stimulus with microwave electromagnetic field and sham exposure. Recognition of both stimuli in a pair was better under the sham exposure conditions but the actual difference was only 5%. It was concluded that early stages of visual information processing are overwhelmingly robust and routine (and adaptively significant) activities, so that the low level 7 Hz modulated electromagnetic field effects exerted upon it are extremely weak.     

Light- and CO2-saturated photosynthesis: enhancement by oxygen

Oxygen may enhance CO₂-saturated photosynthesis in intact leaves, which display the Warburg effect when illuminated at the current atmospheric level of CO₂ and O₂, of about 350 μl l⁻¹ and 21%, respectively. The magnitude of the stimulation depends on irradiance. The K _M(O₂) of the stimulation is 128 μM (10.6% O₂). Maximum enhancement in wheat leaves is 6.1 and 5.3 μmol m⁻² s⁻¹ under 27.9 and 18.7 mW cm⁻², respectively, corresponding to a 25–30% increase in the ribulose 1,5-bisphosphate (RuBP) turnover rate if compared with O₂-free ambient gas phase. The stimulation appears in 5–10 s after a sharp increase in O₂. In response to a decrease in O₂, the new stabilized rate is reached in 5–7 min. The stimulation does not involve any increase in the activity of Rubisco. The effect correlates with increased concentration of RuBP. Oxygen enhances CO₂-saturated photosynthesis by acting as a terminal electron acceptor in the photosynthetic electron transport. The magnitude of the effect may be adopted as an index of the pseudocyclic photophosphorylation in vivo.     

Methods of electroencephalographic signal analysis for detection of small hidden changes

The aim of this study was to select and evaluate methods sensitive to reveal small hidden changes in the electroencephalographic (EEG) signal. Two original methods were considered.     

Estimation of rate constants of the partial reactions of carboxylation of ribulose-1,5-bisphosphate in vivo

An in vivo method for the estimation of kinetic parameters of partial reactions of carboxylation of ribulose 1,5-bisphosphate (RuBP) catalyzed by ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) is described. Rubisco in barley, wheat and bean is different in the ability of its active centers to bind RuBP. The rate constant of the formation of the Rubisco-RuBP complex in these plants at 25 °C is 0.414, 0.245 and 0.660 mM^-1 s^-1, respectively. The rate constant of the reaction of the Rubisco-bound enediol with CO₂ does not differ significantly in barley and wheat, and averages 66 mM^-1 s^-1. Decreased irradiance inhibits Rubisco in two ways: by reducing the concentration of operating catalytic sites and by decreasing the rate constant of binding of RuBP to Rubisco. High concentrations of CO₂ inhibit Rubisco by decreasing the concentration of competent carboxylation centers, without any s ignificant influence upon the rate constants of partial reactions.