Assessment of the cyto- and genotoxic effects of a nanoferromagnetic and a static magnetic field in vivo

The cyto- and genotoxic effects of ferromagnetic nanoparticles (FNs), a static magnetic field (SMF), or their combination were comparatively studied in Ehrlich ascites carcinoma (EAC) cells, bone marrow (BM) erythroid series cells, and peripheral blood lymphocytes at definite time intervals, using the MN test and the DNA-comet assay. The MN test and the DNA-comet assay were used as markers of genotoxic exposure. It has been shown that the limit value of insignificantly expressed changes in the EAC cell architectonics (due to FNs) is an FN concentration of 3 mg/kg. The FN concentration increases in tumor cells, because the expressed blebbing in the cytoplasmic membrane increases the amount of micronuclei and the percentage of DNA in the comet tail in EAC cells, BM erythroid series cells, and peripheral blood lymphocytes. It has also been demonstrated that an SMF alone, as an independent factor, does not produce cytogenotoxic effects on the studied cells. When SMF and FN were used in combination, we could observe the phenomenon of SMF induction by FNs, which manifested itself as an increased total effect of these factors on cells. The MN-test and the DNA-comet assay can be used for assessing the genomic stability of both tumor cells and somatic nonmalignized cells when testing the genotoxic effect of nanomaterials used in the design of vector systems. As was established, the DNA-comet method demonstrated a higher sensitivity in the assessment of the genotoxicity produced by the studied factors.     

Substitutions at the opening of the Rubisco central solvent channel affect holoenzyme stability and CO2/O2 specificity but not activation by Rubisco activase

Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) catalyzes the initial step of carbon metabolism in photosynthesis. The holoenzyme comprises eight large subunits, arranged as a tetramer of dimers around a central solvent channel that defines a fourfold axis of symmetry, and eight small subunits, arranged as two tetramers at the poles of the axis. The phylogenetically divergent small-subunit loops between β-strands A and B form the entrance to the solvent channel. In the green alga Chlamydomonas reinhardtii, Ile-58 from each of the four small-subunit βA–βB loops defines the minimal diameter of the channel opening. To understand the role of the central solvent channel in Rubisco function, directed mutagenesis and transformation of Chlamydomonas were employed to replace Ile-58 with Ala, Lys, Glu, Trp, or three Trp residues (I58W3) to close the entrance to the channel. The I58E, I58K, and I58W substitutions caused only small decreases in photosynthetic growth at 25 and 35 °C, whereas I58W3 had a substantial effect at both temperatures. The mutant enzymes had decreased carboxylation rates, but the I58W3 enzyme had decreases in both carboxylation and CO₂/O₂ specificity. The I58E, I58W, and I58W3 enzymes were inactivated at lower temperatures than wild-type Rubisco, and were degraded at slower rates under oxidative stress. However, these mutant enzymes were activated by Rubisco activase at normal rates, indicating that the structural transition required for carboxylation is not affected by altering the solvent channel opening. Structural dynamics alone may not be responsible for these distant effects on the Rubisco active site.     

Ventricular beat detection in single channel electrocardiograms

Background  

Detection of QRS complexes and other types of ventricular beats is a basic component of ECG analysis. Many algorithms have been proposed and used because of the waves' shape diversity. Detection in a single channel ECG is important for several applications, such as in defibrillators and specialized monitors.     

Dynamics of endogenous cytokinin pools in tobacco seedlings: a modelling approach

Recent advances in cytokinin analysis have made it possible to measure the content of 22 cytokinin metabolites in the tissue of developing tobacco seedlings. Individual types of cytokinins in plants are interconverted to their respective forms by several enzymatic activities (5'-AMP-isopentenyltransferase, adenosine nucleosidase, 5'-nucleotidase, adenosine phosphorylase, adenosine kinase, trans-hydroxylase, zeatin reductase, beta-glucosidase, O-glucosyl transferase, N-glucosyl transferase, cytokinin oxidase). This paper reports modelling and measuring of the dynamics of endogenous cytokinins in tobacco plants grown on media supplemented with isopentenyl adenine (IP), zeatin (Z) and dihydrozeatin riboside (DHZR). Differences in phenotypes generated by the three cytokinins are shown and discussed, and the assumption that substrate concentration drives enzyme kinetics underpinned the construction of a simple mathematical model of cytokinin metabolism in developing seedlings. The model was tested on data obtained from liquid chromatography/tandem mass spectrometry cytokinin measurements on tobacco seedlings grown on Murashige and Skoog agar nutrient medium, and on plants grown in the presence of IP, Z and DHZR. A close match was found between measured and simulated data, especially after a series of iterative parameter searches, in which the parameters were set to obtain the best fit with one of the data sets.     

Slow rate during AF improves ventricular performance by reducing sensitivity to cycle length irregularity

Atrial fibrillation (AF) is characterized by short and irregular ventricular cycle lengths (VCL). While the beneficial effects of heart rate slowing (i.e., the prolongation of VCL) in AF are well recognized, little is known about the impact of irregularity. In 10 anesthetized dogs, R-R intervals, left ventricular (LV) pressure, and aortic flow were collected for >500 beats during fast AF and when the average VCL was prolonged to 75%, 100%, and 125% of the intrinsic sinus cycle length by selective atrioventricular (AV) nodal vagal stimulation. We used the ratio of the preceding and prepreceding R-R intervals (RR(p)/RR(pp)) as an index of cycle length irregularity and assessed its effects on the maximum LV power, the minimum of the first derivative of LV pressure, and the time constant of relaxation by using nonlinear fitting with monoexponential functions. During prolongation of VCL, there was a pronounced decrease in curvature with the formation of a plateau, indicating a lesser dependence on RR(p)/RR(pp). We conclude that prolongation of the VCL during AF reduces the sensitivity of the LV performance parameters to irregularity.     

Inhibitory effects of elevated endogenous cytokinins on nitrate reductase in ipt-expressing tobacco are eliminated by short-term exposure to benzyladenine

Using a novel system for expressing ipt gene from Agrobacterium tumefaciens in tobacco ( Nicotiana tabacum L., cv. Petit Havana SR1), we were able to grow seedlings and teratoma-like tissue with increased content of cytokinins. This material enabled us to investigate new regulatory aspects of nitrate reduction. We grew control plants and plants with elevated cytokinins on MS media, with or without nitrate and benzyladenine (BA). We determined in vitro nitrate reductase (EC 1.6.6.1) activity (NRA) in this plant material. Initially, we found that ipt -expressing plants always displayed lowered levels of NRA when compared to wild-type SR1 plants. We determined that long-term exposure of tobacco plants and tissue to cytokinins caused up to 60% decrease in NRA. Exposure to 40 m M nitrate was able to induce the activity in such plants 3-fold, increasing the activity in SR1 plants more than 5-fold. We were able to restore wild-type levels of NRA in ipt -expressing plants by simultaneous induction of NR with BA and nitrate. Our results suggest that regulation of NR by nitrate and cytokinin is a result of overlaying cytokinin-driven regulatory processes, with those acting in the short-term having a positive effect on NRA, and those acting over extended periods of time having inhibitory effects on NRA.     

Phospholipid and cholesterol levels in the tumor cell plasma membranes with different sensitivity to doxorubicin

The investigation is aimed to study qualitative and quantitative composition of phospholipids, cholesterol content and lipids unsaturation index in plasma membranes of Guerin's carcinoma cells sensitive or resistant to doxorubicin. The comparison of infrared spectra and phospholipids unsaturation index showed that the unsaturation level of fatty acids in plasma membrane from resistant cells was lower than that from sensitive carcinoma cells. 31P-NMR spectroscopy of plasma membranes phospholipids shows the increase of phosphatidylserine and sphingomyeline content in plasma membrane isolated from resistant tumor as compared with sensitive tumor. The levels of phosphatidylcholine and phosphatidylethanolamine were equal in drug-resistant and drug-sensitive carcinoma strains. Changes in plasma membrane from resistant cells result in elevation of plasma membrane microviscosity and phosphatidylserine level increase can suggest the activation of P-glycoprotein-mediated efflux of doxorubicin.     

Interpretation of EMG integral or RMS and estimates of “neuromuscular efficiency” can be misleading in fatiguing contraction

In occupational and sports physiology, reduction of neuromuscular efficiency (NME) and elevation of amplitude characteristics, such as root mean square (RMS) or integral of surface electromyographic (EMG) signals detected during fatiguing submaximal contraction are often related to changes in neural drive. However, there is data showing changes in the EMG integral (I_EMG) and RMS due to peripheral factors. Causes for these changes are not fully understood. On the basis of computer simulation, we demonstrate that lengthening of intracellular action potential (IAP) profile typical for fatiguing contraction could affect EMG amplitude characteristics stronger than alteration in neural drive (central factors) defined by number of active motor units (MUs) and their firing rates. Thus, relation of these EMG amplitude characteristics only to central mechanisms can be misleading. It was also found that to discriminate between changes in RMS or I_EMG due to alterations in neural drive from changes due to alterations in peripheral factors it is better to normalize RMS of EMG signals to the RMS of M-wave. In massive muscles, such normalization is more appropriate than normalization to either peak-to-peak amplitude or area of M-wave proposed in literature.     

The head-fixed behaving rat--procedures and pitfalls

This paper describes experimental techniques with head-fixed, operantly conditioned rodents that allow the control of stimulus presentation and tracking of motor output at hitherto unprecedented levels of spatio-temporal precision. Experimental procedures for the surgery and behavioral training are presented. We place particular emphasis on potential pitfalls using these procedures in order to assist investigators who intend to engage in this type of experiment. We argue that head-fixed rodent models, by allowing the combination of methodologies from molecular manipulations, intracellular electrophysiology, and imaging to behavioral measurements, will be instrumental in combining insights into the functional neuronal organization at different levels of observation. Provided viable behavioral methods are implemented, model systems based on rodents will be complementary to current primate models--the latter providing highest comparability with the human brain, while the former offer hugely advanced methodologies on the lower levels of organization, for example, genetic alterations, intracellular electrophysiology, and imaging.