Respondents of health survey powered by the innovative NURO app exhibit correlations between exercise frequencies and diet habits, and between stress levels and sleep wellness.

Nurosene's NURO app (nurosene.com) is an innovative smartphone application that gathers and analyzes active self-report metrics from users, empowering them with data-driven health machine intelligence. We present the data collected and analyzed from the initial round of participants who responded to a 12-question survey on their life-style and health status. Exploratory results using a variational autoencoder (VAE) suggested that much of the variability of the 12 dimensional data could be accounted for by two approximately uncorrelated latent variables: one pertaining to stress and sleep, and the other pertaining to exercise and diet. Subsequent modeling of the data using exploratory and confirmatory factor analyses (EFAs and CFAs) found that optimal data fits consisted of four factors, namely exercise, diet, stress, and sleep. Covariance values were high between exercise and diet, and between stress and sleep, but much lower between other pairings of non-identical factors. Both EFAs and CFAs provided extra contexts to and quantified the more preliminary VAE observations. Overall, our results significantly reduce the apparent complexity of the response data. This reduction allows for more efficient future stratification and analyses of participants based on simpler latent variables. Our discovery of novel relationships between stress and sleep, and between exercise and diet suggests the possibility of applying predictive analytics in future efforts.

Rotation of sex combs in Drosophila melanogaster requires precise and coordinated spatio-temporal dynamics from forces generated by epithelial cells.

The morphogenesis of sex combs (SCs), a male trait in many species of fruit flies, is an excellent system in which to study the cell biology, genetics and evolution of a trait. In Drosophila melanogaster, where the incipient SC rotates from horizontal to a vertical position, three signal comb properties have been documented: length, final angle and shape (linearity). During SC rotation, in which many cellular processes are occurring both spatially and temporally, it is difficult to distinguish which processes are crucial for which attributes of the comb. We have used a novel approach combining simulations and experiments to uncover the spatio-temporal dynamics underlying SC rotation. Our results indicate that 1) the final SC shape is primarily controlled by the inhomogeneity of initial cell size in cells close to the immature comb, 2) the final angle is primarily controlled by later cell expansion and 3) a temporal sequence of cell expansion mitigates the malformations generally associated with longer rotated SCs. Overall, our work has linked together the morphological diversity of SCs and the cellular dynamics behind such diversity, thus providing important insights on how evolution may affect SC development via the behaviours of surrounding epithelial cells.

Integrin-linked kinase and ELMO2 modulate recycling endosomes in keratinocytes.

The formation of tight cell-cell junctions is essential in the epidermis for its barrier properties. In this tissue, keratinocytes follow a differentiation program tightly associated with their movement from the innermost basal to the outer suprabasal layers, and with changes in their cell-cell adhesion profile. Intercellular adhesion in keratinocytes is mediated through cell-cell contacts, including E-cadherin-based adherens junctions. Although the mechanisms that mediate E-cadherin delivery to the plasma membrane have been widely studied in simple epithelia, this process is less well understood in the stratified epidermis. In this study, we have investigated the role of Engulfment and Cell Motility 2 (ELMO2) and integrin-linked kinase (ILK) in the positioning of E-cadherin-containing recycling endosomes during establishment of cell-cell contacts in differentiating keratinocytes. We now show that induction of keratinocyte differentiation by Ca2+ is accompanied by localization of ELMO2 and ILK to Rab4- and Rab11a-containing recycling endosomes. The positioning of long-loop Rab11a-positive endosomes at areas adjacent to cell-cell contacts is disrupted in ELMO2- or ILK-deficient keratinocytes, and is associated with impaired localization of E-cadherin to cell borders. Our studies show a previously unrecognized role for ELMO2 and ILK in modulation of endosomal positioning, which may play key roles in epidermal sheet maintenance and permeability barrier function.

Network models predict that reduced excitatory fluctuations can give rise to hippocampal network hyper-excitability in MeCP2-null mice.

Rett syndrome is a severe pediatric neurological disorder caused by loss of function mutations within the gene encoding methyl CpG-binding protein 2 (MeCP2). Although MeCP2 is expressed near ubiquitously, the primary pathophysiology of Rett syndrome stems from impairments of nervous system function. One alteration within different regions of the MeCP2-deficient brain is the presence of hyper-excitable network responses. In the hippocampus, such responses exist despite there being an overall decrease in spontaneous excitatory drive within the network. In this study, we generated and used mathematical, neuronal network models to resolve this apparent paradox. We did this by taking advantage of previous mathematical modelling insights that indicated that decreased excitatory fluctuations, but not mean excitatory drive, more critically explain observed changes in hippocampal network oscillations from MeCP2-null mouse slices. Importantly, reduced excitatory fluctuations could also bring about hyper-excitable responses in our network models. Therefore, these results indicate that diminished excitatory fluctuations may be responsible for the hyper-excitable state of MeCP2-deficient hippocampal circuitry.

Inhibitory networks of fast-spiking interneurons generate slow population activities due to excitatory fluctuations and network multistability.

Slow population activities (SPAs) exist in the brain and have frequencies below ~5 Hz. Despite SPAs being prominent in several cortical areas and serving many putative functions, their mechanisms are not well understood. We studied a specific type of in vitro GABAergic, inhibition-based SPA exhibited by C57BL/6 murine hippocampus. We used a multipronged approach consisting of experiment, simulation, and mathematical analyses to uncover mechanisms responsible for hippocampal SPAs. Our results show that hippocampal SPAs are an emergent phenomenon in which the "slowness" of the network is due to interactions between synaptic and cellular characteristics of individual fast-spiking, inhibitory interneurons. Our simulations quantify characteristics underlying hippocampal SPAs. In particular, for hippocampal SPAs to occur, we predict that individual fast-spiking interneurons should have frequency-current (f-I) curves that exhibit a suitably sized kink where the slope of the curve decreases more abruptly in the gamma frequency range with increasing current. We also predict that these interneurons should be well connected with one another. Our mathematical analyses show that the combination of synaptic and intrinsic conditions, as predicted by our simulations, promotes network multistability. Population slow timescales occur when excitatory fluctuations drive the network between different stable network firing states. Since many of the parameters we use are extracted from experiments and subsequent measurements of experimental f-I curves of fast-spiking interneurons exhibit characteristics as predicted, we propose that our network models capture a fundamental operating mechanism in biological hippocampal networks.

Emerging role of ILK and ELMO2 in the integration of adhesion and migration pathways.

Integrins and their associated proteins are essential components of the cellular machinery that modulates adhesion and migration. In particular, integrin-linked kinase (ILK), which binds to the cytoplasmic tail of ß1 integrins, is required for migration in a variety of cell types. We previously identified engulfment and motility 2 (ELMO2) as an ILK-binding protein in epidermal keratinocytes. Recently, we investigated the biological role of the ILK/ELMO2 complexes, and found that they exist in the cytoplasm. ILK/ELMO2 species are recruited by active RhoG to the plasma membrane, where they induce Rac1 activation and formation of lamellipodia at the leading edge of migrating cells. A large number of growth factors and cytokines induce keratinocyte migration. However, we found that formation of RhoG/ELMO2/ILK complexes occurs selectively upon stimulation by epidermal growth factor, but not by transforming growth factor-ß1 or keratinocyte growth factor. Herein we discuss the relevance of these complexes to our understanding of the molecular mechanisms involved in cell migration, as well as their potential functions in morphogenesis and tissue regeneration following injury.

Epidermal growth factor induction of front-rear polarity and migration in keratinocytes is mediated by integrin-linked kinase and ELMO2.

Epidermal growth factor (EGF) is a potent chemotactic and mitogenic factor for epidermal keratinocytes, and these properties are central for normal epidermal regeneration after injury. The involvement of mitogen-activated protein kinases as mediators of the proliferative effects of EGF is well established. However, the molecular mechanisms that mediate motogenic responses to this growth factor are not clearly understood. An obligatory step for forward cell migration is the development of front-rear polarity and formation of lamellipodia at the leading edge. We show that stimulation of epidermal keratinocytes with EGF, but not with other growth factors, induces development of front-rear polarity and directional migration through a pathway that requires integrin-linked kinase (ILK), Engulfment and Cell Motility-2 (ELMO2), integrin ß1, and Rac1. Furthermore, EGF induction of front-rear polarity and chemotaxis require the tyrosine kinase activity of the EGF receptor and are mediated by complexes containing active RhoG, ELMO2, and ILK. Our findings reveal a novel link between EGF receptor stimulation, ILK-containing complexes, and activation of small Rho GTPases necessary for acquisition of front-rear polarity and forward movement.

Expression and analysis of exogenous proteins in epidermal cells.

In this chapter we review protocols for transient transfection of primary keratinocytes. The ability to transfect primary epidermal cells regardless of their differentiation status allows the biochemical and molecular characterization of multiple proteins. We review methods to analyze exogenous protein abundance in transfected keratinocytes by immunoblot and immunoprecipitation. We also present protocols to determine the subcellular distribution of these proteins by indirect immunofluorescence microscopy approaches.

Integrin-linked kinase interactions with ELMO2 modulate cell polarity.

Cell polarization is a key prerequisite for directed migration during development, tissue regeneration, and metastasis. Integrin-linked kinase (ILK) is a scaffold protein essential for cell polarization, but very little is known about the precise mechanisms whereby ILK modulates polarization in normal epithelia. Elucidating these mechanisms is essential to understand tissue morphogenesis, transformation, and repair. Here we identify a novel ILK protein complex that includes Engulfment and Cell Motility 2 (ELMO2). We also demonstrate the presence of RhoG in ILK-ELMO2 complexes, and the localization of this multiprotein species specifically to the leading lamellipodia of polarized cells. Significantly, the ability of RhoG to bind ELMO is crucial for ILK induction of cell polarization, and the joint expression of ILK and ELMO2 synergistically promotes the induction of front-rear polarity and haptotactic migration. This places RhoG-ELMO2-ILK complexes in a key position for the development of cell polarity and forward movement. Although ILK is a component of many diverse multiprotein species that may contribute to cell polarization, expression of dominant-negative ELMO2 mutants is sufficient to abolish the ability of ILK to promote cell polarization. Thus, its interaction with ELMO2 and RhoG is essential for the ability of ILK to induce front-rear cell polarity.

Inhibition dominates in shaping spontaneous CA3 hippocampal network activities in vitro.

We have assessed the balance of excitation and inhibition in in vitro rodent hippocampal slices exhibiting spontaneous, basal sharp waves (bSPWs). A defining signature of a network exhibiting bSPWs is the rise and fall in local field activities with frequencies ranging from 0.5 to 4.5 Hz. This variation of extracellular local field activities manifests at the intracellular level as postsynaptic potentials (PSPs). In correspondence with the local field bSPWs, we consider "sparse" and "synchronous" parts of bSPWs at the intracellular level. We have used intracellular data of bSPW-associated PSPs together with mathematical extraction techniques to quantify the mean and variance of synaptic conductances that a neuron experiences during bSPW episodes. We find that inhibitory conductances dominate in pyramidal cells and in a putative interneuron, and that inhibitory variances are much greater than excitatory ones during synchronous parts of bSPWs. Specifically, we find that there is at least a twofold increase in inhibitory conductance dominance from "sparse" to "synchronous" bSPW states and that this transition is associated with inhibitory fluctuations of greater than 10% of the change in mean inhibitory conductance. On the basis of our findings, we suggest that such inhibitory fluctuations during transition may be a physiological feature of systems expressing such population activities. In summary, our results provide a quantified basis for understanding the interaction of excitatory and inhibitory neuronal subpopulations in bSPW activities.

A simple model for describing the concentration of 212Pb in the atmosphere.

The implementation of the Comprehensive Test Ban Treaty is resulting in the construction of a world-wide system of 80 monitoring stations that will be able to detect air-borne radioactivity, not only from atomic bombs but also from other anthropogenic and natural sources. A prototype monitoring station has been operating since April 1996 in Vancouver, BC, Canada. This station provides daily reports of natural radioactivity, including 7Be and decay products of 220Rn (thoron). Data for 212Pb concentrations have been analyzed over a 6-month period. The concentration is reduced by rainfall, high wind velocity, and low temperatures and it also depends on the wind direction, but atmospheric inversions appeared to have little impact. We present a relatively simple model, which is easy to use and which offers predictive powers that can be applied to other similar environmental situations.