Spatial and temporal control over photoresponsive nanoclusters.

Development of temporal and spatial characteristics of anticipatory postural adjustments during gait initiation in children aged 3-10years.

ABSTRACTAlthough a large number of mouse models have been made to study Alzheimer's disease, only a handful allow experimental control over the location or timing of the protein being used to drive pathology. Other fields have used the Cre and the tamoxifen-inducible CreER driver lines to achieve precise spatial and temporal control over gene deletion and transgene expression, yet these tools have not been widely used in studies of neurodegeneration. Here, we describe two strategies for harnessing the wide range of Cre and CreER driver lines to control expression of disease-associated amyloid precursor protein (APP) in modeling Alzheimer's amyloid pathology. We show that CreER-based spatial and temporal control over APP expression can be achieved with existing lines by combining a Cre driver with a tetracycline-transactivator (tTA)-dependent APP responder using a Cre-to-tTA converter line. We then describe a new mouse line that places APP expression under direct control of Cre recombinase using an intervening lox-stop-lox cassette. Mating this allele with a CreER driver allows both spatial and temporal control over APP expression, and with it, amyloid onset. This article has an associated First Person interview with the first author of the paper.

Temporal and spatial variability of stream waters in Wales, the Welsh borders and part of the West Midlands, UK — 1. Major ion concentrations

Gallium nitride-based light-emitting diodes (LEDs) have revolutionized the lighting industry with their efficient generation of blue and green light. While broad-area (square millimetre) devices have become the dominant LED lighting technology, fabricating LEDs into micro-scale pixels (micro-LEDs) yields further advantages for optical wireless communications (OWC), and for the development of smart-lighting applications such as tracking and imaging. The smaller active areas of micro-LEDs result in high current density operation, providing high modulation bandwidths and increased optical power density. Fabricating micro-LEDs in array formats allows device layouts to be tailored for target applications and provides additional degrees of freedom for OWC systems. Temporal and spatial control is crucial to use the full potential of these micro-scale sources, and is achieved by bonding arrays to pitch-matched complementary metal-oxide-semiconductor control electronics. These compact, integrated chips operate as digital-to-light converters, providing optical signals from digital inputs. Applying the devices as projection systems allows structured light patterns to be used for tracking and self-location, while simultaneously providing space-division multiple access communication links. The high-speed nature of micro-LED array devices, combined with spatial and temporal control, allows many modes of operation for OWC providing complex functionality with chip-scale devices.This article is part of the theme issue ‘Optical wireless communication’.

Posttranscriptional control over rapid development and ciliogenesis in Marsilea

Chemical oscillations are exploited to achieve self-expiring graphical information on paper-based supports with precise temporal and spatial control. Writing and self-erasing processes are chemically activated by exciting nonoscillating Belousov-Zhabotinsky (BZ) solutions infiltrated in cellulose paper filters. Exhausted supports can be reactivated many times by adding new BZ medium. Different parameters can be independently controlled to program mono- or multipaced information storage.

Materials engineered to elicit targeted cellular responses in regenerative medicine must display bioligands with precise spatial and temporal control. Although materials with temporally regulated presentation of bioadhesive ligands using external triggers, such as light and electric fields, have been recently realized for cells in culture, the impact of in vivo temporal ligand presentation on cell-material responses is unknown. Here, we present a general strategy to temporally and spatially control the in vivo presentation of bioligands using cell adhesive peptides with a protecting group that can be easily removed via transdermal light exposure to render the peptide fully active. We demonstrate that non-invasive, transdermal time-regulated activation of cell-adhesive RGD peptide on implanted biomaterials regulates in vivo cell adhesion, inflammation, fibrous encapsulation, and vascularization of the material. This work shows that triggered in vivo presentation of bioligands can be harnessed to direct tissue reparative responses associated with implanted biomaterials.

For studying any event, measurement can never be enough; “control” is required. This means mere passive tracking of the event is insufficient and being able to manipulate it is necessary. To maximize this capability to exert control and manipulate, both spatial and temporal domains need to be jointly accounted for, which has remained an intractable problem at microscopic scales. Simultaneous control of dynamics and position of an observable event requires a holistic combination of spatial and temporal control principles, which gives rise to the field of spatiotemporal control. For this, we present a novel femtosecond pulse-shaping approach. We explain how to achieve spatiotemporal control by spatially manipulating the system through trapping and subsequently or simultaneously exerting temporal control using shaped femtosecond pulses. By leveraging ultrafast femtosecond lasers, the prospect of having temporal control of molecular dynamics increases, and it becomes possible to circumvent the relaxation processes at microscopic timescales. Optical trapping is an exemplary demonstration of spatial control that results in the immobilization of microscopic objects with radiation pressure from a tightly focused laser beam. Conventional single-beam optical tweezers use continuous-wave (CW) lasers for achieving spatial control through photon fluxes, but these lack temporal control knobs. We use a femtosecond high repetition rate (HRR) pulsed laser to bypass this lack of dynamical control in the time domain for optical trapping studies. From a technological viewpoint, the high photon flux requirement of stable optical tweezers necessitates femtosecond pulse shaping at HRR, which has been a barrier until the recent Megahertz pulse shaping developments. Finally, recognizing the theoretical distinction between tweezers with femtosecond pulses and CW lasers is of paramount interest. Non-linear optical (NLO) interactions must be included prima facie to understand pulsed laser tweezers in areas where they excel, like the two-photon-fluorescence-based detection. We show that our theoretical model can holistically address the common drawback of all tweezers. We are able to mitigate the effects of laser-induced heating by balancing this with femtosecond laser-induced NLO effects. An interesting side-product of HRR femtosecond-laser-induced thermal lens is the development of femtosecond thermal lens spectroscopy (FTLS) and its ability to provide sensitive molecular detection.

Chemical biology approaches for protein tagging in mammalian cells.

Tubulin, a central component of the cellular cytoskeleton is critical for the formation of the mitotic spindle in mitosis. Without this dynamic protein, a cell is unable to properly divide. As a result, a number of cancer drugs target tubulin, and prevent the cell from properly undergoing mitosis. A class of drugs known as mitotic inhibitors disrupt tubulin polymerization by acting to stabilize or destabilize tubulin polymer formation. While these drugs are often used to treat disease, they may also be used as biochemical tools to study tubulin polymerization. However, these molecules often lack precise spatial and temporal control, rendering them inadequate to fully study a dynamic process such as tubulin polymerization. Recently, photoisomerizable compounds have gained attention due to their increased spatial and temporal control. Azo‐stilbene compounds are well known to be photoisomerizable between the cis and the trans form at 400‐600nm. This photoisomerization gives azo‐stilbene compounds great power as biochemical tools. Certain mitotic inhibitors have a stilbene backbone, which make azo‐stilbene compounds the ideal starting point for the discovery of photoisomerizeable tubulin polymerization inhibitors. We have developed a library of azo‐stilbenes, which we have supplemented with rationally designed compounds, for the purpose of inhibiting tubulin polymerization. We will discuss the synthesis of these compounds and their activity as novel azo‐stilbene tubulin binders, as well as the implications of our findings for the synthesis of next‐generation compounds to inhibit tubulin polymerization.

Integration of biochemical reaction networks (BRNs) with biosensor platforms has emerged as a technological niche overcoming challenges related to the loss of sensitivity and selectivity in biological media. Optimal operation of BRNs in microfluidics requires control over reaction-diffusion dominated mass transport, heavily influenced by fluidic parameters. In this work, we study and design an on-chip platform combining a programable unique molecular amplification as BRNs with nanoscale biologically sensitive field-effect transistor (BioFET) arrays, which employs a physical diffusion barrier to gain spatial and temporal control over mass transport. Computational and numerical approaches, such as finite element and finite volume methods, were implemented to solve partial differential equations numerically after domain approximation by numerous finite elements. The focus on geometrical optimizations of fluidics is aimed at mass transport to occur with precise spatial and temporal control toward BioFET-arrays. Adopting a 0.5 pM limit-of-detection (LoD) for biochemical monitoring of BRNs via a single-stranded deoxyribonucleic acid (ssDNA) output, we show that it was possible to compartmentalize the mass transport spatiotemporally without crosstalk, which can be of critical advantage for using biosensor arrays in order to realize simplified multiplexed point-of-care biosensors.

Modularity of the Bacterial Cell Cycle Enables Independent Spatial and Temporal Control of DNA Replication

Gene expression systems in Drosophila: a synthesis of time and space

Sporulation in Bacillus subtilis is a simple developmental system in which a single cell undergoes differentiation to two 'sister' cells, namely the prespore and the sporangium. Prespore-specific gene expression is largely dependent on the synthesis of a transcription factor, sigma G. Transcription of spolllG, the gene encoding sigma G, is under precise temporal and spatial control, requiring the products of at least eight genes that are expressed in the pre-divisional cell. Here we show that the product of one of these genes, another sigma factor, sigma F, is by itself sufficient to direct transcription of spolllG in non-sporulating cells. The results indicate that the cell-specificity of prespore gene expression is determined by a mechanism that exerts temporal and spatial control over the activity of sigma F.

Embedded bioprinting is an emerging technology for precise deposition of cell-laden or cell-only bioinks to construct tissue like structures. Bioink is extruded or transferred into a yield stress hydrogel or a microgel support bath allowing print needle motion during printing and providing temporal support for the printed construct. Although this technology has enabled creation of complex tissue structures, it remains a challenge to develop a support bath with user-defined extracellular mimetic cues and their spatial and temporal control. This is crucial to mimic the dynamic nature of the native tissue to better regenerate tissues and organs. To address this, we present a bioprinting approach involving printing of a photocurable viscous support layer and bioprinting of a cell-only or cell-laden bioink within this viscous layer followed by brief exposure to light to partially crosslink the support layer. This approach does not require shear thinning behavior and is suitable for a wide range of photocurable hydrogels to be used as a support. It enables multi-material printing to spatially control support hydrogel heterogeneity including temporal delivery of bioactive cues (e.g. growth factors), and precise patterning of dense multi-cellular structures within these hydrogel supports. Here, dense stem cell aggregates are printed within methacrylated hyaluronic acid-based hydrogels with patterned heterogeneity to spatially modulate human mesenchymal stem cell osteogenesis. This study has significant impactions on creating tissue interfaces (e.g. osteochondral tissue) in which spatial control of extracellular matrix properties for patterned stem cell differentiation is crucial.