Lignin and organic free radicals in maize (Zea mays L.) seeds in response to aflatoxin B1 contamination: an optical and EPR spectroscopic study.

BACKGROUND: Aflatoxin B1 (AFB1 ) is the most dangerous of the mycotoxins that contaminate cereal seeds naturally. A stress lignin formation is linked with the accumulation of reactive oxygen species causing a change in the redox status and formation of stable organic radicals, constituting the first layer of defense. The relationship between AFB1 and changes in lignin organic free radicals in seeds is not known, nor is the part of the seed that is more targeted. Using optical and electron paramagnetic resonance spectroscopy, we investigated AFB1 -induced changes in lignin and organic free radicals in seeds, and whether the inner and outer seed fractions differ in response to increasing AFB1 . RESULTS: Different changes in the content of lignin and free radicals with increasing AFB1 concentrations were observed in the two seed fractions. There was a significant positive linear correlation (R = 0.9923, P = 0.00005) between lignin content and AFB1 concentration in the outer fraction, and no correlation between the lignin content and the AFB1 concentration in the inner fraction. We found a positive correlation between the area of the green spectral emission component (C4) and the AFB1 concentration in the outer fraction. CONCLUSIONS: To the best of our knowledge, the results showed, for the first time, that maize seed fractions respond differently to aflatoxin with regard to their lignin and organic free radical content. Lignin content and (C4) area may be reliable indicators for the screening of lignin changes against AFB1 content in the seeds, and thus for seed protection capacity. © 2021 Society of Chemical Industry.

A perspective view on the nanomotion detection of living organisms and its features.

The insurgence of newly arising, rapidly developing health threats, such as drug-resistant bacteria and cancers, is one of the most urgent public-health issues of modern times. This menace calls for the development of sensitive and reliable diagnostic tools to monitor the response of single cells to chemical or pharmaceutical stimuli. Recently, it has been demonstrated that all living organisms oscillate at a nanometric scale and that these oscillations stop as soon as the organisms die. These nanometric scale oscillations can be detected by depositing living cells onto a micro-fabricated cantilever and by monitoring its displacements with an atomic force microscope-based electronics. Such devices, named nanomotion sensors, have been employed to determine the resistance profiles of life-threatening bacteria within minutes, to evaluate, among others, the effect of chemicals on yeast, neurons, and cancer cells. The data obtained so far demonstrate the advantages of nanomotion sensing devices in rapidly characterizing microorganism susceptibility to pharmaceutical agents. Here, we review the key aspects of this technique, presenting its major applications. and detailing its working protocols.

Multiscale Entropy Analysis: Application to Cardio-Respiratory Coupling.

It is known that in pathological conditions, physiological systems develop changes in the multiscale properties of physiological signals. However, in real life, little is known about how changes in the function of one of the two coupled physiological systems induce changes in function of the other one, especially on their multiscale behavior. Hence, in this work we aimed to examine the complexity of cardio-respiratory coupled systems control using multiscale entropy (MSE) analysis of cardiac intervals MSE (RR), respiratory time series MSE (Resp), and synchrony of these rhythms by cross multiscale entropy (CMSE) analysis, in the heart failure (HF) patients and healthy subjects. We analyzed 20 min of synchronously recorded RR intervals and respiratory signal during relaxation in the supine position in 42 heart failure patients and 14 control healthy subjects. Heart failure group was divided into three subgroups, according to the RR interval time series characteristics (atrial fibrillation (HFAF), sinus rhythm (HFSin), and sinus rhythm with ventricular extrasystoles (HFVES)). Compared with healthy control subjects, alterations in respiratory signal properties were observed in patients from the HFSin and HFVES groups. Further, mean MSE curves of RR intervals and respiratory signal were not statistically different only in the HFSin group (p = 0.43). The level of synchrony between these time series was significantly higher in HFSin and HFVES patients than in control subjects and HFAF patients (p < 0.01). In conclusion, depending on the specific pathologies, primary alterations in the regularity of cardiac rhythm resulted in changes in the regularity of the respiratory rhythm, as well as in the level of their asynchrony.

Cell death parameters as revealed by whole-cell patch-clamp and interval weighted spectra averaging: changes in membrane properties and current frequency of cultured mouse microglial cells induced by glutaraldehyde.

The physiological and biochemical factors that lead to cell death have not been recognized completely. To our knowledge, there are no data on the bioelectric parameters that characterize early period of cell death, as well as on the appearance of related membrane current frequencies. We studied early parameters of glutaraldehyde (GA)-induced cell death, by examining the membrane properties of mouse microglia using the whole-cell patch-clamp technique. In addition, we investigated the GA-induced changes in the membrane current frequency, to see if characteristic frequencies would appear in dying cell. For data analysis, we applied a new approach, an improved multiple moving window length analysis and interval weighted spectra averaging (IWSA). We chose GA for its ability to induce almost instantaneous cell death. The 0.6% GA did not induce changes in the bioelectric membrane properties of microglia. However, the 3% GA caused significant decrease of membrane capacitance and resistance accompanied by the prominent increase in the membrane currents and nearly ohmic current response of microglial cells. These data indicate that 3% GA caused complete loss of the membrane function consequently inducing instantaneous cell death. The membrane function loss was characterized by appearance of the 1.26-4.62 Hz frequency peak in the IWSA spectra, while no significant increase of amplitudes could be observed for cells treated with 0.6% GA. To our knowledge, this is the first record of a frequency associated with complete loss of the membrane function and thus can be considered as an early indicator of cell death.

Optical Nanomotion Detection to Rapidly Discriminate between Fungicidal and Fungistatic Effects of Antifungals on Single-Cell Candida albicans.

Candida albicans is an emerging pathogen that poses a significant challenge due to its multidrug-resistant nature. There are two types of antifungal agents, fungicidal and fungistatic, with distinct mechanisms of action against fungal pathogens. Fungicidal agents kill fungal pathogens, whereas fungistatic agents inhibit their growth. The growth can be restored once the agent is removed and favorable conditions are established. Recognizing this difference is crucial as it influences treatment selection and infection prognosis. We present a technique based on optical nanomotion detection (ONMD) (i.e., observing the movement of the cells using an optical microscope) to discriminate rapidly between fungicidal (caspofungin) and fungistatic (fluconazole) drugs. The technique is based on the change in a yeast cell's nanomotion as a function of time during a two-hour treatment with the antifungal of interest followed by a one-hour growth period. The cells are entrapped in microwells in a microfluidic chip, which allows a quick exchange of growth medium and antifungal agent, enabling ONMD measurements on the same individual cells before and after treatment. This procedure permits to discriminate between fungicidal and fungistatic antifungals in less than 3 h, with single-cell resolution by observing if the nanomotion recovers after removing the treatment and reintroducing growth medium (YPD), or continues to drop. The simplicity of the approach holds promise for further development into a user-friendly device for rapid antifungal susceptibility testing (AFST), potentially being implemented in hospitals and medical centers worldwide in developed and developing countries.

Enhancement in statistical and image analysis for in situ µSXRF studies of elemental distribution and co-localization, using Dioscorea balcanica.

Synchrotron-based X-ray microfluorescence (µSXRF) is an analytical method suitable for in situ investigation of the distribution of micronutrient and macronutrient elements in several-micrometres-thick unstained biological samples, e.g. single cells and tissues. Elements are mapped and quantified at sub-p.p.m. concentrations. In this study the quantity, distribution and grouping/co-localization of various elements have been identified in straight and twisted internodes of the stems of the monocotyledonous climber D. balcanica Kosanin. Three different statistical methods were employed to analyse the macronutrient and micronutrient distributions and co-localization. Macronutrient elements (K, P, Ca, Cl) are distributed homogeneously in both straight and twisted internodes. Micronutrient elements are mostly grouped in the vasculature and in the sclerenchyma cell layer. In addition, co-localization of micronutrient elements is much more prominent in twisted than in straight internodes. These image analyses and statistical methods provided very similar outcomes and could be applied to various types of biological samples imaged by µSXRF.

Two Operational Modes of Cardio-Respiratory Coupling Revealed by Pulse-Respiration Quotient.

Due to the fact that respiratory breath-to-breath and cardiac intervals between two successive R peaks (BBI and RRI, respectively) are not temporally concurrent, in a previous paper, we proposed a method to calculate both the integer and non-integer parts of the pulse respiration quotient (PRQ = BBI/RRI = PRQint + b1 + b2), b1 and b2 being parts of the border RRIs for each BBI. In this work, we study the correlations between BBI and PRQ, as well as those between BBI and mean RRI within each BBI (mRRI), on a group of twenty subjects in four conditions: in supine and standing positions, in combination with spontaneous and slow breathing. Results show that the BBI vs. PRQ correlations are positive; whereas the breathing regime had little or no effect on the linear regression slopes, body posture did. Two types of scatter plots were obtained with the BBI vs. mRRI correlations: one showed points aggregated around the concurrent PRQint lines, while the other showed randomly distributed points. Five out of six of the proposed aggregation measures confirmed the existence of these two cardio-respiratory coupling regimes. We also used b1 to study the positions of R pulses relative to the respiration onsets and showed that they were more synchronous with sympathetic activation. Overall, this method should be used in different pathological states.

Mitochondrial nanomotion measured by optical microscopy.

Nanometric scale size oscillations seem to be a fundamental feature of all living organisms on Earth. Their detection usually requires complex and very sensitive devices. However, some recent studies demonstrated that very simple optical microscopes and dedicated image processing software can also fulfill this task. This novel technique, termed as optical nanomotion detection (ONMD), was recently successfully used on yeast cells to conduct rapid antifungal sensitivity tests. In this study, we demonstrate that the ONMD method can monitor motile sub-cellular organelles, such as mitochondria. Here, mitochondrial isolates (from HEK 293 T and Jurkat cells) undergo predictable motility when viewed by ONMD and triggered by mitochondrial toxins, citric acid intermediates, and dietary and bacterial fermentation products (short-chain fatty acids) at various doses and durations. The technique has superior advantages compared to classical methods since it is rapid, possesses a single organelle sensitivity, and is label- and attachment-free.

Involvement of Na+/K+ pump in fine modulation of bursting activity of the snail Br neuron by 10 mT static magnetic field.

The spontaneously active Br neuron from the brain-subesophageal ganglion complex of the garden snail Helix pomatia rhythmically generates regular bursts of action potentials with quiescent intervals accompanied by slow oscillations of membrane potential. We examined the involvement of the Na(+)/K(+) pump in modulating its bursting activity by applying a static magnetic field. Whole snail brains and Br neuron were exposed to the 10-mT static magnetic field for 15 min. Biochemical data showed that Na(+)/K(+)-ATPase activity increased almost twofold after exposure of snail brains to the static magnetic field. Similarly, (31)P NMR data revealed a trend of increasing ATP consumption and increase in intracellular pH mediated by the Na(+)/H(+) exchanger in snail brains exposed to the static magnetic field. Importantly, current clamp recordings from the Br neuron confirmed the increase in activity of the Na(+)/K(+) pump after exposure to the static magnetic field, as the magnitude of ouabain's effect measured on the membrane resting potential, action potential, and interspike interval duration was higher in neurons exposed to the magnetic field. Metabolic pathways through which the magnetic field influenced the Na(+)/K(+) pump could involve phosphorylation and dephosphorylation, as blocking these processes abolished the effect of the static magnetic field.

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.

Pulse respiration quotient as a measure sensitive to changes in dynamic behavior of cardiorespiratory coupling such as body posture and breathing regime.

Objective: In this research we explored the (homeo)dynamic character of cardiorespiratory coupling (CRC) under the influence of different body posture and breathing regimes. Our tool for it was the pulse respiration quotient (PRQ), representing the number of heartbeat intervals per breathing cycle. We obtained non-integer PRQ values using our advanced Matlab® algorithm and applied it on the signals of 20 healthy subjects in four conditions: supine position with spontaneous breathing (Supin), standing with spontaneous breathing (Stand), supine position with slow (0.1 Hz) breathing (Supin01) and standing with slow (0.1 Hz) breathing (Stand01). Main results: Linear features of CRC (in PRQ signals) were dynamically very sensitive to posture and breathing rhythm perturbations. There are obvious increases in PRQ mean level and variability under the separated and joined influence of orthostasis and slow (0.1 Hz) breathing. This increase was most pronounced in Stand01 as the state of joint influences. Importantly, PRQ dynamic modification showed greater sensitivity to body posture and breathing regime changes than mean value and standard deviation of heart rhythm and breathing rhythm. In addition, as a consequence of prolonged supine position, we noticed the tendency to integer quantization of PRQ (especially after 14 min), in which the most common quantization number was 4:1 (demonstrated in other research reports as well). In orthostasis and slow breathing, quantization can also be observed, but shifted to other values. We postulate that these results manifest resonance effects induced by coupling patterns from sympathetic and parasympathetic adjustments (with the second as dominant factor). Significance: Our research confirms that cardiorespiratory coupling adaptability could be profoundly explored by precisely calculated PRQ parameter since cardiorespiratory regulation in healthy subjects is characterized by a high level of autonomic adaptability (responsiveness) to posture and breathing regime, although comparisons with pathological states has yet to be performed. We found Stand01 to be the most provoking state for the dynamic modification of PRQ (cardiorespiratory inducement). As such, Stand01 has the potential of using for PRQ tuning by conditioning the cardiorespiratory autonomic neural networks, e.g., in the cases where PRQ is disturbed by environmental (i.e., microgravity) or pathologic conditions.

Influence of silicon on polymerization process during lignin synthesis. Implications for cell wall properties.

Silicon (Si) is considered a beneficial element for plants, mostly accumulating in cell walls, where its location and content are primed by the chemistry and structure of lignin. It is unrevealed how Si interacts with the process of lignin formation in the CWs. We studied, in an in vitro system, the interaction of SiO2 with the peroxidase-catalyzed polymerization of a lignin monomer into the lignin model compound, imitating conditions of the last step of lignin formation. FTIR and fluorescence spectroscopy and microscopy showed that Si is bound to the final polymer, and the structure of the Si-DHP differs from pure DHP. Fluorescence spectroscopy showed that Si does not bind to the monomers, so Si probably inhibits the formation of the larger lignin fragments, as evidenced by HPLC-DAD, by binding to dimmers formed during DHP synthesis. The structural changes of the polymer are related to the changed proportion of the fractions of various MW. The enzyme catalyzing DHP synthesis was not inhibited by Si. HRP activity was increased in presence of Si except for 6 mM Si. This may indicate that the complex formed with Si and short oligomers activates the enzyme, and prevents the formation of the large fragments.

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.

Coupling changes in cortical and pontine sigma and theta frequency oscillations following monoaminergic lesions in rat.

PURPOSE: Sigma and theta frequency electroencephalogram (EEG) oscillations exhibit substantial and well-recognized shifts with transitions across sleep and wake states. We aimed in this study to test the changes in coupling between these characteristic oscillations of non-rapid-eye-movement (NREM)/rapid-eye-movement (REM) sleep within and between cortical and pontine EEGs following monoaminergic lesion, by using the Pearson's product-moment correlation coefficients. METHODS: Experiments were performed in 14 adult, male Sprague Dawley rats chronically instrumented for sleep recording. We lesioned the dorsal raphe nucleus axon terminals in four rats using PCA neurotoxin (p-chloroamphetamine; Sigma-Aldrich, MO) administered as two intraperitoneal (IP) injections (6 mg/kg) 24 h apart. Lesioning of locus coeruleus axon terminals was performed in five rats using DSP-4 neurotoxin (N-2-chloroethyl-N-ethyl-2-bromobenzilamine; Sigma-Aldrich, MO) in a single IP dose of 50 mg/kg. RESULTS & CONCLUSIONS: Our previous study [Saponjic et al., Physiol Behav 90:1-10, 2007] demonstrated that these systemically induced monoaminergic lesions failed to produce significant changes in sleep/wake distribution from control conditions. The present study, by using spectral analysis and by examining the Pearson's correlation coefficients and their approximate probability density (APD) distribution profiles in control and lesion condition, demonstrates significant augmentation of the sigma/theta coupling strength, an inversion of cortical sigma/theta coupling direction and emergence of an additional sigma/theta coupling "mode" specific to the post-lesion state only within the cortex. By using the Pearson's correlation coefficients and their APD profiles, instead of classical sleep/wake distribution analysis, as a measure of direction and strength of sigma/theta coupling within and between cortex and pons, we were able to uncover the impact of a tonically decreased level of brain monoamines as altered strength and mode of coupling between sigma and theta oscillations. Specifically, a new mode of sigma/theta coupling emerged following lesion, which was specific to NREM sleep, suggests that loss of monoaminergic signaling interferes with NREM sleep consolidation. Our results also indicate an importance of monoamines in control of the sleep spindle and theta rhythm generators.

Quantification of compression wood severity in tracheids of Pinus radiata D. Don using confocal fluorescence imaging and spectral deconvolution.

Confocal fluorescence microscopy was used to examine the spectral characteristics of lignin autofluorescence in secondary cell walls of normal and compression wood from Pinus radiata. Using UV excitation, fluorescence spectra of normal and compression wood sections showed significant differences, especially in the outer secondary cell wall of tracheids, with a shift in maxima from violet to blue wavelengths between normal and compression wood. A comparison of normal wood, mild and severe compression wood, showed that the wavelength shift was intermediate in the mild compression wood compared to the severe compression wood, thus offering the possibility of quantifying the severity by measuring ratios of fluorescence at violet and blue wavelengths. Fluorescence induced by blue light, rather than UV, was less well differentiated amongst wood types. Spectral deconvolution indicated the presence of a minimum of five discrete lignin fluorophores in the cell walls of both normal and compression wood tracheids. Comparison with lignin model compounds suggest that the wavelength shift may correspond in part to increased levels of p-hydroxy type lignin in the compression wood samples. The combination of confocal fluorescence imaging and related spectral deconvolution therefore offers a novel technique for characterising cell wall lignin in situ.

Modeling EEG fractal dimension changes in wake and drowsy states in humans--a preliminary study.

Aim of this preliminary study was to examine and compare topographic distribution of Higuchi's fractal dimension (FD, measure of signal complexity) of EEG signals between states of relaxed wakefulness and drowsiness, as well as their FD differences. The experiments were performed on 10 healthy individuals using a fourteen-channel montage. An explanation is offered on the causes of the detected FD changes. FD values of 60s records belonging to wake (Hori's stage 1) and drowsy (Hori's stages 2-4) states were calculated for each channel and each subject. In 136 out of 140 epochs an increase in FD was obtained. Relationship between signal FD and its relative alpha amplitude was mathematically modeled and we quantitatively demonstrated that the increase in FD was predominantly due to a reduction in alpha activity. The model was generalized to include other EEG oscillations. By averaging FD values for each channel across 10 subjects, four clusters (O2O1; T6P4T5P3; C3F3F4C4F8F7; T4T3) for the wake and two clusters (O2O1P3T6P4T5; C3C4F4F3F8T4T3F7) for the drowsy state were statistically verified. Topographic distribution of FD values in wakefulness showed a lateral symmetry and a partial fronto-occipital gradient. In drowsiness, a reduction in the number of clusters was detected, due to regrouping of channels T3, T4, O1 and O2. Topographic distribution of absolute FD differences revealed largest values at F7, O1 and F3. Reorganization of channel clusters showed that regionalized brain activity, specific for wakefulness, became more global by entering into drowsiness. Since the global increase in FD during wake-to-drowsy transition correlated with the decrease of alpha power, we inferred that increase of EEG complexity may not necessarily be an index of brain activation.

The effect of pH on the activity of soluble peroxidase in needles of Serbian spruce (Picea omorika (Panc.) Purkinye): application of a mathematical model.

We studied the dependence of peroxidase (POD) activity on pH in crude extract of Picea omorika (Panc.) Purkinye needles and in its acidic and basic fractions, obtained by ion exchange chromatography. Nonlinear regression was applied on the activity data with pH as the explaining variable, using the Levenberg-Marquardt algorithm. Studying crude extract at three different temperatures, the shape of the simulated activity/pH dependences indicated an existence of two components, which was confirmed by mathematical modeling. The kinetic parameters Act0, KEH and KEOH of both components are presented. The curves and pH optima shifted under increasing temperatures towards lower pH values, which was verified after decomposition. Nonlinear regression detected the presence of two components for both fractions, and there is no considerable difference between their pH optima. Our results show for the first time that the sum of components, each described by the mathematical model employed, can be used to explain the complex pH-related POD activity in the extract with two or more enzyme forms simultaneously active.

Slow 0.1 Hz Breathing and Body Posture Induced Perturbations of RRI and Respiratory Signal Complexity and Cardiorespiratory Coupling.

Objective: We explored the physiological background of the non-linear operating mode of cardiorespiratory oscillators as the fundamental question of cardiorespiratory homeodynamics and as a prerequisite for the understanding of neurocardiovascular diseases. We investigated 20 healthy human subjects for changes using electrocardiac RR interval (RRI) and respiratory signal (Resp) Detrended Fluctuation Analysis (DFA, α1RRI, α2RRI, α1Resp, α2Resp), Multiple Scaling Entropy (MSERRI1-4, MSERRI5-10, MSEResp1-4, MSEResp5-10), spectral coherence (CohRRI-Resp), cross DFA (ρ1 and ρ2) and cross MSE (XMSE1-4 and XMSE5-10) indices in four physiological conditions: supine with spontaneous breathing, standing with spontaneous breathing, supine with 0.1 Hz breathing and standing with 0.1 Hz breathing. Main results: Standing is primarily characterized by the change of RRI parameters, insensitivity to change with respiratory parameters, decrease of CohRRI-Resp and insensitivity to change of in ρ1, ρ2, XMSE1-4, and XMSE5-10. Slow breathing in supine position was characterized by the change of the linear and non-linear parameters of both signals, reflecting the dominant vagal RRI modulation and the impact of slow 0.1 Hz breathing on Resp parameters. CohRRI-Resp did not change with respect to supine position, while ρ1 increased. Slow breathing in standing reflected the qualitatively specific state of autonomic regulation with striking impact on both cardiac and respiratory parameters, with specific patterns of cardiorespiratory coupling. Significance: Our results show that cardiac and respiratory short term and long term complexity parameters have different, state dependent patterns. Sympathovagal non-linear interactions are dependent on the pattern of their activation, having different scaling properties when individually activated with respect to the state of their joint activation. All investigated states induced a change of α1 vs. α2 relationship, which can be accurately expressed by the proposed measure-inter-fractal angle θ. Short scale (α1 vs. MSE1-4) and long scale (α2 vs. MSE5-10) complexity measures had reciprocal interrelation in standing with 0.1 Hz breathing, with specific cardiorespiratory coupling pattern (ρ1 vs. XMSE1-4). These results support the hypothesis of hierarchical organization of cardiorespiratory complexity mechanisms and their recruitment in ascendant manner with respect to the increase of behavioral challenge complexity. Specific and comprehensive cardiorespiratory regulation in standing with 0.1 Hz breathing suggests this state as the potentially most beneficial maneuver for cardiorespiratory conditioning.

Single yeast cell nanomotions correlate with cellular activity.

Living single yeast cells show a specific cellular motion at the nanometer scale with a magnitude that is proportional to the cellular activity of the cell. We characterized this cellular nanomotion pattern of nonattached single yeast cells using classical optical microscopy. The distribution of the cellular displacements over a short time period is distinct from random motion. The range and shape of such nanomotion displacement distributions change substantially according to the metabolic state of the cell. The analysis of the nanomotion frequency pattern demonstrated that single living yeast cells oscillate at relatively low frequencies of around 2 hertz. The simplicity of the technique should open the way to numerous applications among which antifungal susceptibility tests seem the most straightforward.

Kinetic parameters for thermal inactivation of soluble peroxidase from needles of Serbian spruce Picea omorika (Pancic) Purkyne.

Thermal inactivation of peroxidase (POD) in an extract of Picea omorika (Pancic) Purkyne needles initiated by heat treatment was studied. This is the first study of this kind on a conifer species. Non-linear regression analysis was applied on the inactivation rate data, combining Mitscherlich and Arrhenius equations, treating time and temperature simultaneously as explaining variables. We determined the inactivation rate constant k, the Arrhenius energy of inactivation E and the remaining activity C(min) for the crude extract and for separated acidic and basic enzyme fractions, as well as for individual isoenzymes separated electrophoretically. A comparison of inactivation parameters for acidic and basic fractions shows that the thermal inactivation rate of the basic fraction is higher. The obtained value of inactivation energy for crude extract was between the values for acidic and basic isoenzyme fractions. One of the three analysed individual isoenzymes was characterised by a lower inactivation rate constant and higher inactivation energy. Another isoenzyme showed considerably higher level of remaining activity compared to the others, which identified it as the most resistant to high temperatures. The acquired values of Arrhenius energy of inactivation for POD in crude extract were intermediate, considering a range of POD values for various other plant species.