Measuring and Modeling the Effect of Audio on Human Focus in Everyday Environments Using Brain-Computer Interface Technology.

The goal of this study was to investigate the effect of audio listened to through headphones on subjectively reported human focus levels, and to identify through objective measures the properties that contribute most to increasing and decreasing focus in people within their regular, everyday environment. Participants (N = 62, 18-65 years) performed various tasks on a tablet computer while listening to either no audio (silence), popular audio playlists designed to increase focus (pre-recorded music arranged in a particular sequence of songs), or engineered soundscapes that were personalized to individual listeners (digital audio composed in real-time based on input parameters such as heart rate, time of day, location, etc.). Audio stimuli were delivered to participants through headphones while their brain signals were simultaneously recorded by a portable electroencephalography headband. Participants completed four 1-h long sessions at home during which different audio played continuously in the background. Using brain-computer interface technology for brain decoding and based on an individual's self-report of their focus, we obtained individual focus levels over time and used this data to analyze the effects of various properties of the sounds contained in the audio content. We found that while participants were working, personalized soundscapes increased their focus significantly above silence (p = 0.008), while music playlists did not have a significant effect. For the young adult demographic (18-36 years), all audio tested was significantly better than silence at producing focus (p = 0.001-0.009). Personalized soundscapes increased focus the most relative to silence, but playlists of pre-recorded songs also increased focus significantly during specific time intervals. Ultimately we found it is possible to accurately predict human focus levels a priori based on physical properties of audio content. We then applied this finding to compare between music genres and revealed that classical music, engineered soundscapes, and natural sounds were the best genres for increasing focus, while pop and hip-hop were the worst. These insights can enable human and artificial intelligence composers to produce increases or decreases in listener focus with high temporal (millisecond) precision. Future research will include real-time adaptation of audio for other functional objectives beyond affecting focus, such as affecting listener enjoyment, drowsiness, stress and memory.

Serotonin-dependent kinetics of feeding bursts underlie a graded response to food availability in C. elegans.

Animals integrate physiological and environmental signals to modulate their food uptake. The nematode C. elegans, whose food uptake consists of pumping bacteria from the environment into the gut, provides excellent opportunities for discovering principles of conserved regulatory mechanisms. Here we show that worms implement a graded feeding response to the concentration of environmental bacteria by modulating a commitment to bursts of fast pumping. Using long-term, high-resolution, longitudinal recordings of feeding dynamics under defined conditions, we find that the frequency and duration of pumping bursts increase and the duration of long pauses diminishes in environments richer in bacteria. The bioamine serotonin is required for food-dependent induction of bursts as well as for maintaining their high rate of pumping through two distinct mechanisms. We identify the differential roles of distinct families of serotonin receptors in this process and propose that regulation of bursts is a conserved mechanism of behaviour and motor control.

Durable spatiotemporal surveillance of Caenorhabditis elegans response to environmental cues.

Animal response to changes in environmental cues is a complex dynamical process that occurs at diverse molecular and cellular levels. To gain a quantitative understanding of such processes, it is desirable to observe many individuals, subjected to repeatable and well defined environmental cues over long time periods. Here we present WormSpa, a microfluidic system where worms are individually confined in optimized chambers. We show that worms in WormSpa are neither stressed nor starved, and in particular exhibit pumping and egg-laying behaviors equivalent to those of freely behaving worms. We demonstrate the applicability of WormSpa for studying stress response and physiological processes. WormSpa is simple to make and easy to operate, and its design is modular, making it straightforward to incorporate available microfluidic technologies. We expect that WormSpa would open novel avenues of research, hitherto impossible or impractical.

Nucleocytoplasmic transport: a thermodynamic mechanism.

The nuclear pore supports molecular communication between cytoplasm and nucleus in eukaryotic cells. Selective transport of proteins is mediated by soluble receptors, whose regulation by the small GTPase Ran leads to cargo accumulation in, or depletion from, the nucleus, i.e., nuclear import or nuclear export. We consider the operation of this transport system by a combined analytical and experimental approach. Provocative predictions of a simple model were tested using cell-free nuclei reconstituted in Xenopus egg extract, a system well suited to quantitative studies. We found that accumulation capacity is limited, so that introduction of one import cargo leads to egress of another. Clearly, the pore per se does not determine transport directionality. Moreover, different cargo reach a similar ratio of nuclear to cytoplasmic concentration in steady-state. The model shows that this ratio should in fact be independent of the receptor-cargo affinity, though kinetics may be strongly influenced. Numerical conservation of the system components highlights a conflict between the observations and the popular concept of transport cycles. We suggest that chemical partitioning provides a framework to understand the capacity to generate concentration gradients by equilibration of the receptor-cargo intermediary.

Protein dynamics in individual human cells: experiment and theory.

A current challenge in biology is to understand the dynamics of protein circuits in living human cells. Can one define and test equations for the dynamics and variability of a protein over time? Here, we address this experimentally and theoretically, by means of accurate time-resolved measurements of endogenously tagged proteins in individual human cells. As a model system, we choose three stable proteins displaying cell-cycle-dependant dynamics. We find that protein accumulation with time per cell is quadratic for proteins with long mRNA life times and approximately linear for a protein with short mRNA lifetime. Both behaviors correspond to a classical model of transcription and translation. A stochastic model, in which genes slowly switch between ON and OFF states, captures measured cell-cell variability. The data suggests, in accordance with the model, that switching to the gene ON state is exponentially distributed and that the cell-cell distribution of protein levels can be approximated by a Gamma distribution throughout the cell cycle. These results suggest that relatively simple models may describe protein dynamics in individual human cells.

Diffusion of anionic and neutral GFP derivatives through plasmodesmata in epidermal cells of Nicotiana benthamiana.

Plasmodesmata (Pd) are trans-wall membrane channels that permit cell-to-cell transport of metabolites and other small molecules, proteins, RNAs, and signaling molecules. The transport of cytoplasmic soluble macromolecules is a function of the electrochemical gradient between adjacent cells, the number of Pd per interface between adjacent cells, Stokes radius (R(S)), area of the cytoplasmic annulus, and channel length. The size of the largest molecule that can pass through Pd defines the Pd size exclusion limit. However, since the shape and size of a molecule determines its capacity to diffuse through pores or tubes, R(S) is a better measure. Relatively small changes in R(S) can cause large differences in the mobility of molecular probes, particularly if the pore size is close to that of the probe. In addition, as the dimensions of a macromolecule approach that of the channel, membrane charge effects may become important. We employed quantitative tools and molecular modeling to measure the apparent coefficient of conductivity of Pd, C(Pd), for the non-targeted transport of macromolecules. This method allowed us to examine the influence of protein charge and R(S) on C(Pd) in Nicotiana benthamiana. The C(Pd) of modified green fluorescent proteins (GFPs) of different sizes but with the same charge as native GFP and of a more negatively charged derivative were determined. We found that the C(Pd) of cytoplasmic soluble GFP and cytoplasmic forms of modified GFP were the most strongly correlated with R(S) and that the apparent aberrant increase in C(Pd) of a negatively charged GFP derivative was, at least in part, the result of the charge effect on R(S).

Reversibility in nucleocytoplasmic transport.

Nucleocytoplasmic exchange of proteins and RNAs is mediated by receptors that usher their cargo through the nuclear pores. Peptide localization signals on each cargo determine the receptors with which it will interact. Those interactions are normally regulated by the small GTPase Ran. Hydrolysis of GTP provides the chemical energy required to create a bona fide thermodynamic pump that selectively and directionally accumulates its substrates across the nuclear envelope. A common perception is that cargo delivery is irreversible, e.g., a protein imported to the nucleus does not return to the cytoplasm except perhaps via a specific export receptor. Quantitative measurements using cell-free nuclei reconstituted in Xenopus egg extract show that nuclear accumulation follows first-order kinetics and reaches steady state at a level that follows a Michaelis-Menten function of the cytoplasmic cargo concentration. This saturation suggests that receptor-mediated translocation across the nuclear pore occurs bidirectionally. The reversibility of accumulation was demonstrated directly by exchange of the cytosolic medium and by fluorescence recovery after photobleaching. Based on our results, we offer a simple biophysical model that predicts the observed behavior. A far-reaching consequence is that the nuclear localization signal dictates the fate of a protein population rather than that of the individual molecules that bear it, which remain free to shuttle back and forth. This implies an open communication between the nucleus and cytoplasm and a ubiquitous mechanism for signaling in both directions.

CD8 kinetically promotes ligand binding to the T-cell antigen receptor.

The mechanism of CD8 cooperation with the TCR in antigen recognition was studied on live T cells. Fluorescence correlation measurements yielded evidence of the presence of two TCR and CD8 subpopulations with different lateral diffusion rate constants. Independently, evidence for two subpopulations was derived from the experimentally observed two distinct association phases of cognate peptide bound to class I MHC (pMHC) tetramers and the T cells. The fast phase rate constant ((1.7 +/- 0.2) x 10(5) M(-1) s(-1)) was independent of examined cell type or MHC-bound peptides' structure. Its value was much faster than that of the association of soluble pMHC and TCR ((7.0 +/- 0.3) x 10(3) M(-1) s(-1)), and close to that of the association of soluble pMHC with CD8 ((1-2) x 10(5) M(-1) s(-1)). The fast binding phase disappeared when CD8-pMHC interaction was blocked by a CD8-specific mAb. The latter rate constant was slowed down approximately 10-fold after cells treatment with methyl-beta-cyclodextrin. These results suggest that the most efficient pMHC-cell association route corresponds to a fast tetramer binding to a colocalized CD8-TCR subpopulation, which apparently resides within membrane rafts: the reaction starts by pMHC association with the CD8. This markedly faster step significantly increases the probability of pMHC-TCR encounters and thereby promotes pMHC association with CD8-proximal TCR. The slow binding phase is assigned to pMHC association with a noncolocalized CD8-TCR subpopulation. Taken together with results of cytotoxicity assays, our data suggest that the colocalized, raft-associated CD8-TCR subpopulation is the one capable of inducing T-cell activation.