Stochastic Spatial Heterogeneity in Activities of H+-ATP-ases in Electrically Connected Plant Cells Decreases Threshold for Cooling-Induced Electrical Responses

H+-ATP-ases, which support proton efflux through the plasma membrane, are key molecular transporters for electrogenesis in cells of higher plants. Initial activities of the transporters can influence the thresholds of generation of electrical responses induced by stressors and modify other parameters of these responses. Previously, it was theoretically shown that the stochastic heterogeneity of individual cell thresholds for electrical responses in a system of electrically connected neuronal cells can decrease the total threshold of the system (“diversity-induced resonance”, DIR). In the current work, we tested a hypothesis about decreasing the thresholds of generation of cooling-induced electrical responses in a system of electrically connected plant cells with increasing stochastic spatial heterogeny in the initial activities of H+-ATP-ases in these cells. A two-dimensional model of the system of electrically connected excitable cells (simple imitation of plant leaf), which was based on a model previously developed in our works, was used for the present investigation. Simulation showed that increasing dispersion in the distribution of initial activities of H+-ATP-ases between cells decreased the thresholds of generation of cooling-induced electrical responses. In addition, the increasing weakly influenced the amplitudes of electrical responses. Additional analysis showed two different mechanisms of the revealed effect. The increasing spatial heterogeneity in activities of H+-ATP-ases induced a weak positive shift of the membrane potential at rest. The shift decreased the threshold of electrical response generation. However, the decreased threshold induced by increasing the H+-ATP-ase activity heterogeneity was also observed after the elimination of the positive shift. The result showed that the “DIR-like” mechanism also participated in the revealed effect. Finally, we showed that the standard deviation of the membrane potentials before the induction of action potentials could be used for the estimation of thresholds of cooling-induced plant electrical responses. Thus, spatial heterogeneity in the initial activities of H+-ATP-ases can be a new regulatory mechanism influencing the generation of electrical responses in plants under actions of stressors.

Sequence and primer independent stochastic heterogeneity in PCR amplification efficiency revealed by single molecule barcoding

The polymerase chain reaction (PCR) is one of the most widely used techniques in molecular biology. In combination with High Throughput Sequencing (HTS), PCR is widely used to quantify transcript abundance for RNA-seq and especially in the context of analysis of T cell and B cell receptor repertoires. In this study, we combine molecular DNA barcoding with HTS to quantify PCR output from individual target molecules. Our results demonstrate that the PCR process exhibits very significant unexpected heterogeneity, which is independent of the sequence of the primers or target, and independent of bulk experimental conditions. The mechanistic origin of this heterogeneity is not clear, but simulations suggest that it must derive from inherited differences between different DNA molecules within the reaction. The results illustrate that single molecule barcoding is important in order to derive reproducible quantitative results from any protocol which combines PCR with HTS.

SAMPLING CONCRETE BY TOO SMALL ELEMENTS; WHAT SHOULD WE DO TO GET RELIABLE INFORMATION?

Macroscopically heterogeneous materials like concrete are generally sampled by too small, i.e., subrepresentative elements that can be either of 2D (section images) or of 3D nature (specimens). Based on scientific notions, like stochastic heterogeneity and structure-sensitivity, which are at the very heart of materials science and stereology, the paper demonstrates biases in obtained information to be generally inevitable when derived from such sub-representative designs. Only reliable comparison studies can be performed under the condition that the linear size of samples and of minimum structural dimensions (resulting from observation resolution) are maintained as fixed proportions of the relevant representative area and/or volume elements. This is demonstrated by three case studies.