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.

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.

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.

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.

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.

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.

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.

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.

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.

Generalized Poincaré plots analysis of heart period dynamics in different physiological conditions: Trained vs. untrained men.

BACKGROUND: Recently we proposed a new method called generalized Poincaré plot (gPp) analysis which gave a new insight into the pattern of neurocaridac control. In this study we examined potential of gPp method to reveal changes in cardiac neural control in young athletes during three conditions: supine rest, running and relaxation, with respect to untrained subjects. METHODS: This method is based on the quantification of Pearson's correlation coefficients r(j, k), between symmetrical (j = k) and asymmetrical summed j previous and k following RR intervals up to the 100th order (j,k≤100). RESULTS: Differences between groups were obtained at all levels of this analysis. The main result is the significant difference of NAI, normalized index of asymmetry, between groups in running, which was originated in different positions of local maxima of r(j, k). Compared with untrained subjects, these findings indicate modified neural control and altered intrinsic heart rate behavior in athletes which are related to some kind of memory mechanism between RR intervals. CONCLUSION: Obtained results provide great potential of gPp method analysis in the recognition of changes in neurocardiac control in healthy subjects. Further studies are needed for identification of altered cardiac regulatory mechanisms whose background may be useful in the evaluation of genesis of athletes neurocardiovascular pathology.

Differentiation of Heart Failure Patients by the Ratio of the Scaling Exponents of Cardiac Interbeat Intervals.

Heart failure (HF) is one of the most frequent heart diseases. It is usually characterized with structural and functional cardiac abnormalities followed by dysfunction of autonomic cardiac control. Current methods of heartbeat interval analysis are not capable to differentiate HF patients and some new differentiation of HF patients could be useful in the determination of the direction of their treatment. In this study, we examined potential of the ratio of the short-term and long-term scaling exponents (α 1 and α 2) to separate HF patients with similar level of reduced cardiac autonomic nervous system control and with no significant difference in age, left ventricular ejection fraction (LVEF) and NYHA class. Thirty-five healthy control subjects and 46 HF patients underwent 20 min of continuous supine resting ECG recording. The interbeat interval time series were analyzed using standardized power spectrum analysis, detrended fluctuation analysis method and standard Poincaré plot (PP) analysis with measures of asymmetry of the PP. Compared with healthy control group, in HF patients linear measures of autonomic cardiac control were statistically significantly reduced (p < 0.05), heart rate asymmetry was preserved (C up > C down, p < 0.01), and long-term scaling exponent α 2 was significantly higher. Cluster analysis of the ratio of short- and long-term scaling exponents showed capability of this parameter to separate four clusters of HF patients. Clusters were determined by interplay of presence of short-term and long-term correlations in interbeat intervals. Complementary measure, commonly accepted ratio of the PP descriptors, SD2/SD1, showed tendency toward statistical significance to separate HF patients in obtained clusters. Also, heart rate asymmetry was preserved only in two clusters. Finally, a multiple regression analysis showed that the ratio α 1/α 2 could be used as an integrated measure of cardiac dynamic with complex physiological background which, besides spectral components as measures of autonomic cardiac control, also involves breathing frequency and mechanical cardiac parameter, left ventricular ejection fraction.

New complexity measures reveal that topographic loops of human alpha phase potentials are more complex in drowsy than in wake.

A number of measures, stemming from nonlinear dynamics, exist to estimate complexity of biomedical objects. In most cases they are appropriate, but sometimes unconventional measures, more suited for specific objects, are needed to perform the task. In our present work, we propose three new complexity measures to quantify complexity of topographic closed loops of alpha carrier frequency phase potentials (CFPP) of healthy humans in wake and drowsy states. EEG of ten adult individuals was recorded in both states, using a 14-channel montage. For each subject and each state, a topographic loop (circular directed graph) was constructed according to CFPP values. Circular complexity measure was obtained by summing angles which directed graph edges (arrows) form with the topographic center. Longitudinal complexity was defined as the sum of all arrow lengths, while intersecting complexity was introduced by counting the number of intersections of graph edges. Wilcoxon's signed-ranks test was used on the sets of these three measures, as well as on fractal dimension values of some loop properties, to test differences between loops obtained in wake vs. drowsy. While fractal dimension values were not significantly different, longitudinal and intersecting complexities, as well as anticlockwise circularity, were significantly increased in drowsy. Graphical abstract An example of closed topographic carrier frequency phase potential (CFPP) loops, recorded in one of the subjects in the wake (A) and drowsy (C) states. Lengths of loop graph edges, r(c j, c j + 1), plotted against the series of EEG channels with decreasing CFPP values, c j , in the wake (B) and drowsy (D) states. Conventional fractal analysis did not reveal any difference between them; therefore, three new complexity measures were introduced.

Slower EEG alpha generation, synchronization and "flow"-possible biomarkers of cognitive impairment and neuropathology of minor stroke.

BACKGROUND: We investigated EEG rhythms, particularly alpha activity, and their relationship to post-stroke neuropathology and cognitive functions in the subacute and chronic stages of minor strokes. METHODS: We included 10 patients with right middle cerebral artery (MCA) ischemic strokes and 11 healthy controls. All the assessments of stroke patients were done both in the subacute and chronic stages. Neurological impairment was measured using the National Institute of Health Stroke Scale (NIHSS), whereas cognitive functions were assessed using the Montreal Cognitive Assessment (MoCA) and MoCA memory index (MoCA-MIS). The EEG was recorded using a 19 channel EEG system with standard EEG electrode placement. In particular, we analyzed the EEGs derived from the four lateral frontal (F3, F7, F4, F8), and corresponding lateral posterior (P3, P4, T5, T6) electrodes. Quantitative EEG analysis included: the group FFT spectra, the weighted average of alpha frequency (αAVG), the group probability density distributions of all conventional EEG frequency band relative amplitudes (EEG microstructure), the inter- and intra-hemispheric coherences, and the topographic distribution of alpha carrier frequency phase potentials (PPs). Statistical analysis was done using a Kruskal-Wallis ANOVA with a post-hoc Mann-Whitney U two-tailed test, and Spearman's correlation. RESULTS: We demonstrated transient cognitive impairment alongside a slower alpha frequency (αAVG) in the subacute right MCA stroke patients vs. the controls. This slower alpha frequency showed no amplitude change, but was highly synchronized intra-hemispherically, overlying the ipsi-lesional hemisphere, and inter-hemispherically, overlying the frontal cortex. In addition, the disturbances in EEG alpha activity in subacute stroke patients were expressed as a decrease in alpha PPs over the frontal cortex and an altered "alpha flow", indicating the sustained augmentation of inter-hemispheric interactions. Although the stroke induced slower alpha was a transient phenomenon, the increased alpha intra-hemispheric synchronization, overlying the ipsi-lesional hemisphere, the increased alpha F3-F4 inter-hemispheric synchronization, the delayed alpha waves, and the newly established inter-hemispheric "alpha flow" within the frontal cortex, remained as a permanent consequence of the minor stroke. This newly established frontal inter-hemispheric "alpha flow" represented a permanent consequence of the "hidden" stroke neuropathology, despite the fact that cognitive impairment has been returned to the control values. All the detected permanent changes at the EEG level with no cognitive impairment after a minor stroke could be a way for the brain to compensate for the lesion and restore the lost function. DISCUSSION: Our study indicates slower EEG alpha generation, synchronization and "flow" as potential biomarkers of cognitive impairment onset and/or compensatory post-stroke re-organizational processes.

Prediction of fish catch in the Danube River based on long-term variability in environmental parameters and catch statistics.

The effects of physical factors on fish catch in the Serbian part of the Danube River were studied for period of six decades. The data on total catch for the Danube River from river kilometre 845 to river kilometre 1433 were collected from Statistical Office of the Republic of Serbia, while water level and water temperature data were collected from 16 water gauge stations along the investigated part of the Danube River for the period 1948-2009. Cross-correlation functions have been used to analyse the functional connection between Danube water level, water temperature and fish catch while ARMA model which combines cyclic (deterministic) and random (stochastic) components of the analysed sequences was used for the forecasts. The cross-correlation function showed negative correlation between water level and temperature as well as between water temperature and catch and positive correlation between water level and catch. The Danube water level and catch were coherent at the periods of 2.06, 4.13, 6.2, 10.33, 20.66years, while the cross correlation function between these time series did not show phase lag. The results of reconstruction and forecast of water level, temperature, and catch of fish in the Danube River, obtained by summing the cyclic and stochastic components, was used for the forecast till 2029. In 2016, seven years after, the initial forecasts were made, validity of the model was checked by obtaining data for water temperature, water level and fish catch in the Danube River for the period 2010-2015. Model gave the best prediction for water temperature; average standard error was 1.6 times higher for predicted value than for model value while for fish catch and water level they were 1.96 and 4.97, respectively. Methods used in this work could be powerful tool for prediction of fish catch and serve as the basis for better fisheries management.

REM sleep disorder following general anesthesia in rats.

Postoperative sleep disorders, particularly the REM sleep disorder, may have a significant deleterious impact on postoperative outcomes and may contribute to the genesis of certain delayed postoperative complications. We have followed the effect of distinct anesthesia regimens (ketamine/diazepam vs. pentobarbital) over 6days following the induction of a stable anesthetized state in adult male Wistar rats, chronically instrumented for sleep recording. In order to compare the effect of both anesthetics in the physiological controls vs. the rats with impaired pedunculopontine tegmental nucleus (PPT) cholinergic innervation, during the operative procedure for the implantation of EEG and EMG electrodes, the bilateral PPT lesion was conducted using ibotenic acid (IBO). We have followed in particular post-anesthesia REM sleep. Our results show the distinct EEG microstructure of the motor cortex during the different stable anesthetized states, and their distinct impact on post-anesthesia REM sleep. In contrast to pentobarbital anesthesia, the ketamine/diazepam anesthesia potentiated the long-lasting post-anesthesia REM statewith higher muscle tone (REM1) vs. REM state with atonia (REM2). Whereas both anesthesias prolonged the post-anesthesia REM sleep duration, the long-term prolongation of the REM1 state was demonstrated only after the ketamine/diazepam anesthesia, first due to the increased number of REM1 episodes, and then due to the prolonged REM1 episodes duration. On the other hand, whereas both anesthetic regimens abolished the prolonged post-anesthesia REM/REM1 sleep and the EEG microstructure disorder during REM sleep, only the pentobarbital abolished the increased NREM/REM/NREM transitions, caused by the PPT lesion. In addition, in the PPT lesioned rats, the ketamine/diazepam anesthesia decreased the Wake/NREM/Wake transitions while the pentobarbital anesthesia decreased the Wake/REM/Wake transitions. Our present study suggests pentobarbital anesthesia as being highly beneficial for post-anesthesia REM sleep in the physiological condition as well as during PPT cholinergic neuropathology.

Uncoupling of cardiac and respiratory rhythm in atrial fibrillation.

Rearranged origin of heart rhythm in patients with atrial fibrillation (AF) influences the regulation of the heart and consequently the respiratory rhythm, and the bidirectional interaction of these rhythms not documented. Hence, we examined coupling of the RR interval and the respiration (Resp) signal by coherence, Granger causality and the cross-sample entropy method of time series analysis in patients with AF and a healthy control group. In healthy subjects, the influence of respiration on cardiac rhythm was found as increased coherence at the breathing frequency (BF) range, significantly stronger interaction and synchrony from Resp to RR than from RR to Resp. On the contrary, in patients with AF, coherence at BF diminished, there were no causal interactions between signals in both directions, which resulted in equally great asynchrony between them. In AF, the absence of full functionality of the sinoatrial node, as an integrator of neural cardiac control, resulted in diminished vagal modulation of heart periods and consequently impaired bidirectional cardio-respiratory interaction.

Generalized Poincaré Plots-A New Method for Evaluation of Regimes in Cardiac Neural Control in Atrial Fibrillation and Healthy Subjects.

Classical Poincaré plot is a standard way to measure nonlinear regulation of cardiovascular control. In our work we propose a generalized form of Poincaré plot where we track correlation between the duration of j preceding and k next RR intervals. The investigation was done in healthy subjects and patients with atrial fibrillation, by varying j,k ≤ 100. In cases where j = k, in healthy subjects the typical pattern was observed by "paths" that were substituting scatterplots and that were initiated and ended by loops of Poincaré plot points. This was not the case for atrial fibrillation patients where Poincaré plot had a simple scattered form. More, a typical matrix of Pearson's correlation coefficients, r(j,k), showed different positions of local maxima, depending on the subject's health condition. In both groups, local maxima were grouped into four clusters which probably determined specific regulatory mechanisms according to correlations between the duration of symmetric and asymmetric observed RR intervals. We quantified matrices' degrees of asymmetry and found that they were significantly different: distributed around zero in healthy, while being negative in atrial fibrillation. Also, Pearson's coefficients were higher in healthy than in atrial fibrillation or in signals with reshuffled intervals. Our hypothesis is that by this novel method we can observe heart rate regimes typical for baseline conditions and "defense reaction" in healthy subjects. These data indicate that neural control mechanisms of heart rate are operating in healthy subjects in contrast with atrial fibrillation, identifying it as the state of risk for stress-dependent pathologies. Regulatory regimes of heart rate can be further quantified and explored by the proposed novel method.

Age-related disorders of sleep and motor control in the rat models of functionally distinct cholinergic neuropathology.

We studied the impact of aging during sleep in the rat models of Alzheimer's (AD) and Parkinson's (PD) disease cholinergic neuropathology to determine the possible different and earlier onset of age-related sleep disorder during the neurodegenerative diseases vs. healthy aging. We used the bilateral nucleus basalis (NB) and pedunculopontine tegmental nucleus (PPT) lesioned rats as the in vivo models of functionally distinct cholinergic neuropathology, and we followed the impact of aging on sleep architecture, the electroencephalographic (EEG) microstructure and motor control across sleep/wake states. Our results have shown for the first time that the earliest signs of aging during distinct cholinergic neuropathology were expressed through a different and topographically specific EEG microstructure during rapid eye movement sleep (REM). EEG delta amplitude attenuation within the sensorimotor cortex (SMCx) during REM was the earliest sign of aging in the NB lesion. EEG sigma amplitude augmentation within the motor cortex (MCx) during REM was the earliest sign of aging in the PPT lesion. In addition, aging was differently expressed through the SMCx drive alterations, but it was commonly expressed through the MCx drive alterations during all sleep/wake states. Our study provided evidence of distinct REM sleep disorders and sleep state related cortical drives as the signs of aging onset during functionally distinct cholinergic neuropathologies (NB lesion vs. PPT lesion).

Anti-cancer effects of cerium oxide nanoparticles and its intracellular redox activity.

Data on medical applications of cerium oxide nanoparticles CeO2 (CONP) are promising, yet information regarding their action in cells is incomplete and there are conflicting reports about in vitro toxicity. Herein, we have studied cytotoxic effect of CONP in several cancer and normal cell lines and their potential to change intracellular redox status. The IC50 was achieved only in two of eight tested cell lines, melanoma 518A2 and colorectal adenocarcinoma HT-29. Self-propagating room temperature method was applied to produce CONP with an average crystalline size of 4 nm. The results confirmed presence of Ce(3+) and O(2-) vacancies. The induction of cell death by CONP and the production of reactive oxygen species (ROS) were analyzed by flow-cytometry. Free radicals related antioxidant capacity of the cells was studied by the reduction of stable free radical TEMPONE using electron spin resonance spectroscopy. CONP showed low or moderate cytotoxicity in cancer cell lines: adenocarcinoma DLD1 and multi-drug resistant DLD1-TxR, non-small cell lung carcinoma NCI-H460 and multi-drug resistant NCI-H460/R, while normal cell lines (keratinocytes HaCaT, lung fetal fibroblasts MRC-5) were insensitive. The most sensitive were 518A2 melanoma and HT-29 colorectal adenocarcinoma cell lines, with the IC50 values being between 100 and 200 µM. Decreased rate of TEMPONE reduction and increased production of certain ROS species (peroxynitrite and hydrogen peroxide anion) indicates that free radical metabolism, thus redox status was changed, and antioxidant capacity damaged in the CONP treated 518A2 and HT-29 cells. In conclusion, changes in intracellular redox status induced by CONP are partly attributed to the prooxidant activity of the nanoparticles. Further, ROS induced cell damages might eventually lead to the cell death. However, low inhibitory potential of CONP in the other human cell lines tested indicates that CONP may be safe for human usage in industry and medicine.

Aging induced cortical drive alterations during sleep in rats.

We followed the impact of healthy aging on cortical drive during sleep in rats by using the corticomuscular coherence (CMC). We employed the chronic electrodes implantation for sleep recording in adult, male Wistar rats, and followed the aging impact during sleep from 3 to 5.5 months age. We have analyzed the sleep/wake states architecture, and the sleep/wake state related EEG microstructure and CMCs. We evidenced the topographically distinct impact of aging on sleep/wake states architecture within the sensorimotor (SMCx) vs. motor cortex (MCx) from 4.5 to 5.5 months age. Healthy aging consistently altered only the SMCx sleep/wake states architecture, and increased the delta and beta CMCs through both cortical drives during Wake, but only through the MCx drive during REM. According to the delta and beta CMCs values, aging impact through the SMCx drive was opposite, but it was convergent through the MCx drive during Wake vs. REM, and there was a dual and inverse mode for the motor control during REM.