Spatial Control Research Articles

CRISPR perfect adaptation for robust control of cellular immune and apoptotic responses.

A central challenge in the quest for precise gene regulation within mammalian cells is the development of regulatory networks that can achieve perfect adaptation-where outputs consistently return to a set baseline post-stimulus. Here, we present such a system that leverages the CRISPR activation (CRISPRa) and anti-CRISPR proteins as two antithetic elements to establish perfect adaptation in mammalian cells and dynamically regulate gene expression. We demonstrate that this system can maintain stable expression levels of target genes in the face of external perturbations, thus providing a robust platform for biological applications. The versatility of our system is further showcased through its integration with endogenous regulatory mechanisms in T cells, such as the NF-κB-mediated immune response, and its ability to program apoptosis responses for precise spatial and temporal control of cellular growth and death. This study not only advances our understanding of gene regulation in mammalian cells but also opens new avenues for therapeutic intervention, particularly in diseases characterized by dysregulated gene expression.

Asymmetric-Backed Multi-Frequency Ultrasonic Transducer for Conformal Tumor Ablation.

Minimally invasive ultrasound ablation transducers have been widely studied. However, conventional designs are limited by the single working frequency, restricting their conformal ablation ability (i.e., ablation size and shape controllability). New multi-frequency ultrasonic transducer design method is proposed based on the asymmetric backing layer, which divides the transducer into non-backing-layer region (i.e., front-piezoelectric region) and backing-layer region (i.e., front-piezoelectric-backing region) with multiple local thickness mode resonant frequencies. Ablation zone can be controlled by exciting the local resonance within or between the regions, and its control flexibility is further enhanced by driven under a frequency modulation signal. Experiments and calculations are combined for verifying the proposal. The fabricated transducer with a Y-direction asymmetric backing layer shows five resonances, with two in each region and one resonance excited in both regions. Spatial ultrasound emission is demonstrated by acoustic measurements. Tissue ablation experiments verified spatial ablation zone control, and frequency modulation driving method enables the spatial transition of ablation zone from one region to the other, generating different ablation sizes and shapes. Finally, patient-specific simulations verified the effectiveness of conformal ablation. The proposed transducer enables flexible control of ablation zone. This study demonstrates a new method for conformal tumor ablation.

Transient Photoactivation of Anionophores by Using Redshifted Fast-Relaxing Azobenzenes.

Photo-regulated transmembrane ionophores enable spatial and temporal control over activity, offering promise as targeted therapeutics. Key to such applications is control using bio-compatible visible light. Herein, we report red-shifted azobenzene-derived synthetic anionophores that use amber or red light to trigger (E)-(Z) photoisomerisation and activation of transmembrane chloride transport. We demonstrate that by tuning the thermal half-life of the more active, but thermodynamically unstable, Z isomer to relax on the timescale of minutes, transient activation of ion transport can be achieved by activating with solely with visible light and deactivating by thermal relaxation.

Gene Electrotransfer via Conductivity-Clamped Electric Field Focusing Pivots Sensori-Motor DNA Therapeutics: "A Spoonful of Sugar Helps the Medicine Go Down".

Viral vectors and lipofection-based gene therapies have dispersion-dependent transduction/transfection profiles that thwart precise targeting. The study describes the development of focused close-field gene electrotransfer (GET) technology, refining spatial control of gene expression. Integration of fluidics for precise delivery of "naked" plasmid deoxyribonucleic acid (DNA)in sucrose carrier within the focused electric field enables negative biasing of near-field conductivity ("conductivity-clamping"-CC), increasing the efficiency of plasma membrane molecular translocation. This enables titratable gene delivery with unprecedently low charge transfer. The clinic-ready bionics-derived CC-GET device achieved neurotrophin-encoding miniplasmid DNA delivery to the cochlea to promote auditory nerve regeneration; validated in deafened guinea pig and cat models, leading to improved central auditory tuning with bionics-based hearing. The performance of CC-GET is evaluated in the brain, an organ problematic for pulsed electric field-based plasmid DNA delivery, due to high required currents causing Joule-heating and damaging electroporation. Here CC-GET enables safe precision targeting of gene expression. In the guinea pig, reporter expression is enabled in physiologically critical brainstem regions, and in the striatum (globus pallidus region) delivery of a red-shifted channelrhodopsin and a genetically-encoded Ca2+sensor, achieved photoactivated neuromodulation relevant to the treatment of Parkinson's Disease and other focal brain disorders.

Unveiling the Transition From Paleolake Lisan to Dead Sea Through the Analysis of Lake Paleoshorelines and Radiometric Dating of Fossil Stromatolites

AbstractTo date, the most complete paleolake‐level reconstructions for the late Pleistocene water bodies that once occupied the Dead Sea depression have been based on the combination of dating of lake sediments and terrestrial materials. However, despite these major accomplishments, there is still limited spatial control regarding the water levels, suggesting some degree of uncertainty concerning the magnitude and rate of lake‐level changes. Here, we re‐examine the late Pleistocene lake‐level changes in the Dead Sea during the transition from paleolake Lisan to the present‐day Dead Sea. We rely on systematic dating of fossil stromatolites including 84 radiocarbon and 15 U‐series ages, stable‐isotope measurements, paleobiology, high‐resolution topography, and numerical modeling to assess lake‐level changes. Our results indicate that the highstand of paleolake Lisan was of shorter duration and the transition between Lake Lisan and the Dead Sea occurred at least 5 Kyrs earlier than previously indicated. By refining the timeline and accuracy of lake‐level positions during the transition paleolake Lisan—Dead Sea, our study offers new insights into the regional and local paleo‐climatic conditions during the last glacial period in this region.

Spatial Control of 2D Nanomaterial Electronic Properties Using Chiral Light Beams.

Single-layer two-dimensional (2D) nanomaterials exhibit physical and chemical properties which can be dynamically modulated through out-of-plane deformations. Existing methods rely on intricate micromechanical manipulations (e.g., poking, bending, rumpling), hindering their widespread technological implementation. We address this challenge by proposing an all-optical approach that decouples strain engineering from micromechanical complexities. This method leverages the forces generated by chiral light beams carrying orbital angular momentum (OAM). The inherent sense of twist of these beams enables the exertion of controlled torques on 2D monolayer materials, inducing tailored strain. This approach offers a contactless and dynamically tunable alternative to existing methods. As a proof-of-concept, we demonstrate control over the conductivity of graphene transistors using chiral light beams, showcasing the potential of this approach for manipulating properties in future electronic devices. This optical control mechanism holds promise in enabling the reconfiguration of devices through optically patterned strain. It also allows broader utilization of strain engineering in 2D nanomaterials for advanced functionalities in next-generation optoelectronic devices and sensors.

Land Use Change Pattern of Change and Strategy Directions for Spatial Utilization Control in Bogor Regency

The designation of Bogor Regency as a National Activity Center encourages regional development, a very high population leads to rapid development and uncontrolled land use change. Therefore, spatial modeling based on land change prediction is needed that can be used as a basis for directing spatial utilization control policies. The purpose of this research is to know and analyze the pattern of land use change and analyze the direction of spatial use control strategies in Bogor Regency. The research was conducted in Bogor Regency. The analysis used ArcGIS 10.8 Overlay and IDRISI Selva Edition and ISM v. 2.3. The results of the analysis of land use change in between 1997 and 2010, 87.98% of land use remained unchanged, while 12.01% changed. From 2010 to 2023, 73.16% remained unchanged, and 26.84% changed. The predominant land change pattern was from Dry Land Agriculture to Dry Land Agriculture-Rice Fields, while the least common was Open Land to Dry Land Agriculture and back to Open Land. By 2036, expected increases include settlements, shrubs, and Open Land, with decreases anticipated in dryland agriculture, forests, and paddy fields. The alignment of land use with the RTRW spatial plan shows 58.68% aligned, 23.85% transitional, and 17.46% misaligned. It is recommended to maintain land use aligned with the spatial plan, utilize transitional areas, and revise the RTRW for areas that are permanently misaligned. Strategies to control spatial utilization include enhancing technical training, improving infrastructure, aligning staff with organizational needs, and refining spatial planning policies and dispute resolution.

Rapid and Controllable Multilayer Cell Sheet Assembly via Biodegradable Nanochannel Membranes

AbstractThe ability to precisely arrange and control the assembly of diverse cell types into intricate 3D structures remains a critical challenge in tissue engineering. Herein, a versatile and programmable 3D cell sheet assembly is described technology by developing a biodegradable nanochannel (BNC) membrane to fulfill this unmet need. This membrane, hierarchically assembled from 2D nanomaterial aggregates, exhibits both exceptional fluid permeability and rapid biodegradation under physiological conditions. The unique properties of the BNC membrane enable precise spatial and temporal control over cell assembly, facilitating the creation of complex 3D cellular architectures. The BNC membrane is integrated with a programmable negative‐pressure‐based cell assembly strategy to form single and multicellular 3D sheets in a highly controllable manner. To demonstrate the feasibility and translatability of this technology in the field of tissue engineering approaches to screen stem cell‐derived therapeutics with “core–shell” macrophage‐fibroblast multicellular patterns and treat murine diabetic skin wounds via scaffold‐free 3D adipose‐derived mesenchymal stem cell (ADMSC) sheets are devised. In summary, the results demonstrate that the BNC membrane‐based 3D cell sheet assembly approach significantly advances current tissue engineering capabilities, offering substantial potential for both regenerative medicine applications and the development of physiologically relevant disease models.

Spatial analysis of multi-frequency SAW beams excited by slanted IDTs on ZnO-SiC heterostructures

Slanted (or fan-shaped) interdigital transducers (IDTs) with broadband response allow the selective excitation of surface acoustic waves (SAWs) with narrow beam widths and pathways controlled by the excitation frequency. Such SAW-based spatial and frequency control is important for applications in microfluidics, as well as in emerging applications in semiconductor nanostructures. In this contribution, we generate both Rayleigh and Sezawa modes with slanted IDTs on a 4H-SiC substrate coated with a piezoelectric ZnO film. We directly measure the phase wavefronts of narrow SAW beams in the 1200–1260 MHz frequency bandwidth using high-resolution ( <1μm ) optical interferometry, and discuss the mechanisms that directly affect the propagation direction and phase profiles of the SAW beams. The combination of multimodal and multi-frequency SAW delay lines provides rich opportunities for SAW-based control of low-dimensional systems, such as electrons in epitaxial graphene or spin centers near the surface of the SiC substrates.

Dynamic control of cascaded four-wave mixing via re-configurable dispersion.

We integrate a spatial light modulator-based dispersion controller into a cascaded four-wave mixing (CFWM) system. By tuning the group delay dispersion (GDD) and fourth-order dispersion (FOD) terms, we control the CFWM phase matching and demonstrate an output bandwidth tuning of over 3.3×. At the maximum bandwidth, our system covers the telecommunications S-, C-, and L-bands (1466-1641 nm) with an average output power of 300 mW, which is contained in 52 individual lines spaced 374 GHz apart. This method represents a reconfigurable alternative to photonic crystal fibers for dispersion engineering and allows for the use of step-index fiber and customizable power spectral density (PSD) profiles.

Schutz- und Sorgepraktiken: Wie LSBT-Organisationen in Deutschland Safer Spaces für ihre Arbeit mit queeren Asylbewerber:innen und Geflüchteten adaptieren

Abstract. Since the “summer of migration” in 2015, LGBT organizations have been actively involved in migration work, particularly in creating safer spaces for queer asylum seekers and refugees. This involvement marks a shift as these organizations previously had limited engagement with queer people of color. German LGBT organizations focus on providing support, security, and community for queer asylum seekers and refugees, drawing upon the concept of safer spaces. While existing literature addresses LGBT organizations in development and the challenges faced by queer asylum seekers, there is limited research on the creation of safer spaces. Research suggests a predominant focus on spatial separation as a means of ensuring safety. This article aims to fill this research gap by exploring how German LGBT organizations are adapting and institutionalizing the concept of safer spaces. It investigates the role of spatial separation, control mechanisms, and potential essentialist and cultural biases in shaping safer spaces. The study contextualizes this within the framework of homonationalism in Germany, followed by a theoretical exploration of safer spaces and an empirical analysis of how LGBT organizations conceptualize them. Drawing on expert interviews with spokespersons of German LGBT organizations and an extensive analysis of their websites, my study shows the ways in which the LGBT organizations construct themselves as saviors of vulnerable queer asylum seekers and migrants, while constructing the non-queer asylum seekers and migrants as a threat through othering. Following the logics of these safer space concepts, spatial separation appears as the ultimate solution to offer safety to queer migrants and refugees. In contrast, I also observe alternative safer spaces offered by LGBT organizations, which, in addition to their purely protective function, also create spaces for caring practices and peer-to-peer empowerment through storytelling.

The Paradox Between Economic Gain and Environmental Sustainability: A Case of Small Island Tourism in Pari Island, Indonesia

As an important economic sector in Indonesia, tourism, particularly for small islands, should shift towards sustainable management practices. This study examines possible sustainable tourism management practices in Pari Island, one of Indonesia???s small islands with both a diverse ecosystem and high annual tourist flux. Tourism activities in Pari Island have resulted in environmental impacts, and implementing sustainable tourism has therefore become challenging. This study used a comprehensive assessment framework called DPSIR (Driver‐Pressure‐State‐Impact‐Response) to identify factors that triggered environmental problems and assess their implications for sustainable tourism management. A questionnaire method was used to collect survey data from 317 residents and 143 tourists. Water quality samples were collected from 15 sampling points (11 groundwater wells and 4 seawater locations) and tested via the APHA‐9221‐B method to determine the coliform bacteria content as a proxy for tourism pressure on the environment. Results showed that the small island tourism was built upon economic activities that trigger a range of environmental pressures (e.g., increased population, tourists, and lodging), with consequences for tourism-related resource exploitation, spatial changes, and contamination of groundwater and seawater. Implementing the DPSIR approach would ensure adequate responses to the main problems of small island tourism, such as creation of regulations informed by tourism???s carrying capacity, an open???close system for use of coastal and marine areas, increased public and tourist awareness, and legal assurance over spatial control.

Engineering programmable material-to-cell pathways via synthetic notch receptors to spatially control differentiation in multicellular constructs

Synthetic Notch (synNotch) receptors are genetically encoded, modular synthetic receptors that enable mammalian cells to detect environmental signals and respond by activating user-prescribed transcriptional programs. Although some materials have been modified to present synNotch ligands with coarse spatial control, applications in tissue engineering generally require extracellular matrix (ECM)-derived scaffolds and/or finer spatial positioning of multiple ligands. Thus, we develop here a suite of materials that activate synNotch receptors for generalizable engineering of material-to-cell signaling. We genetically and chemically fuse functional synNotch ligands to ECM proteins and ECM-derived materials. We also generate tissues with microscale precision over four distinct reporter phenotypes by culturing cells with two orthogonal synNotch programs on surfaces microcontact-printed with two synNotch ligands. Finally, we showcase applications in tissue engineering by co-transdifferentiating fibroblasts into skeletal muscle or endothelial cell precursors in user-defined micropatterns. These technologies provide avenues for spatially controlling cellular phenotypes in mammalian tissues.

Boosting Microfluidic Enzymatic Cascade Reactions with pH-Responsive Polymersomes by Spatio-Chemical Activity Control.

Microfluidic flow reactors permit the implementation of sensitive biocatalysts in polymeric environments (e.g., hydrogel dots), mimicking nature through the use of diverse microstructures within defined confinements. However, establishing complex hybrid structures to mimic biological processes and functions under continuous flow with optimal utilization of all components involved in the reaction process represents a significant scientific challenge. To achieve spatial, chemical, and temporal control for any microfluidic application, compartmentalization is required, as well as the unification of different sensitive compartments in the reaction chamber for the microfluidic flow design. This study presents a self-regulating microfluidic system fabricated by a sequential photostructuring process with an intermediate chemical process step to realize pH-sensitive hybrid structures for the fabrication of a microfluidic double chamber reactor for controlled enzymatic cascade reaction (ECR). The key point is the adaptation and retention of the function of pH-responsive horseradish peroxidase-loaded polymersomes in a microfluidic chip under continuous flow. ECR is successfully triggered and controlled by an interplay between glucose oxidase-converted glucose, the membrane state of pH-responsive polymersomes, and other parameters (e.g., flow rate and fluid composition). This study establishes a promising noninvasive regulatory platform for extended spatio-chemical control of current and future ECR and other cascade reaction systems.

Zonal Patterning of Extracellular Matrix and Stromal Cell Populations Along a Perfusable Cellular Microchannel.

The spatial organization of biophysical and biochemical cues in the extracellular matrix (ECM) in concert with reciprocal cell-cell signaling is vital to tissue patterning during development. However, elucidating the role an individual microenvironmental factor plays using existing in vivo models is difficult due to their inherent complexity. In this work, we have developed a microphysiological system to spatially pattern the biochemical, biophysical, and stromal cell composition of the ECM along an epithelialized 3D microchannel. This technique is adaptable to multiple hydrogel compositions and scalable to the number of zones patterned. We confirmed that the methodology to create distinct zones resulted in a continuous, annealed hydrogel with regional interfaces that did not hinder the transport of soluble molecules. Further, the interface between hydrogel regions did not disrupt microchannel structure, epithelial lumen formation, or media perfusion through an acellular or cellularized microchannel. Finally, we demonstrated spatially patterned tubulogenic sprouting of a continuous epithelial tube into the surrounding hydrogel confined to local regions with stromal cell populations, illustrating spatial control of cell-cell interactions and signaling gradients. This easy-to-use system has wide utility for modeling three-dimensional epithelial and endothelial tissue interactions with heterogeneous hydrogel compositions and/or stromal cell populations to investigate their mechanistic roles during development, homeostasis, or disease.

SPRING licenses S1P-mediated cleavage of SREBP2 by displacing an inhibitory pro-domain

Site-one protease (S1P) conducts the first of two cleavage events in the Golgi to activate Sterol regulatory element binding proteins (SREBPs) and upregulate lipogenic transcription. S1P is also required for a wide array of additional signaling pathways. A zymogen serine protease, S1P matures through autoproteolysis of two pro-domains, with one cleavage event in the endoplasmic reticulum (ER) and the other in the Golgi. We recently identified the SREBP regulating gene, (SPRING), which enhances S1P maturation and is necessary for SREBP signaling. Here, we report the cryo-EM structures of S1P and S1P-SPRING at sub-2.5 Å resolution. SPRING activates S1P by dislodging its inhibitory pro-domain and stabilizing intra-domain contacts. Functionally, SPRING licenses S1P to cleave its cognate substrate, SREBP2. Our findings reveal an activation mechanism for S1P and provide insights into how spatial control of S1P activity underpins cholesterol homeostasis.

Spatial Differentiation and Environmental Controls of Land Consolidation Effectiveness: A Remote Sensing-Based Study in Sichuan, China

The increasing global population is leading to a decline in cropland per person, aggravating food security challenges. The global implementation of land consolidation (LC) has created new farmland and increased productivity. LC is a critical strategy in China for ensuring food security and gaining significant government support. This article investigates the impact of LC on farmland productivity in Sichuan Province in 2020. We utilize time series remote sensing data to analyze LC’s impact on farmland capacity. This study uses Sentinel and Landsat satellite data to calculate CumVI and assesses the LC project’s spatiotemporal evolution. To evaluate LC’s effectiveness, we create indexes for yield level and stability and employ Getis-Ord Gi* to identify spatial differentiation in LC’s impact. GeoDetector and GWR examine the impact of natural factors like elevation, slope, soil organic carbon, and rainfall on the effectiveness of LC. The research results show that: (1) After the implementation of LC, 55.51% of the project areas experienced significant improvements in agricultural productivity; the average increase rate of yield level is 7.74%; and the average increase rate of yield stability is 12.40%. Overall, LC is significant for improving farmland capacity. (2) The effectiveness of LC exhibits spatial differences and correlations in different areas. The main location for high-value agglomeration of yield levels is Nanchong City, while the northern part of Guangyuan City primarily hosts low-value agglomeration areas. (3) Natural conditions influence LC’s effectiveness. In terms of affecting the yield level of LC, the driving factors from high to low are SOC, elevation, slope, and rainfall. In terms of affecting the yield stability of LC, the driving factors, from high to low, are elevation, SOC, slope, and rainfall. LC’s effectiveness is influenced by different natural conditions that have different effects.

Fabrication and Characterization of Collagen–Magnetic Particle Composite Microbeads for Targeted Cell Adhesion and Proliferation

The integration of the biocompatibility of collagen and the remote-control ability of magnetic elements serves as both a cell scaffold and an actuator. We studied the preparation, characterization, and potential applications of collagen–magnetic particle composite microbeads (CMPMBs). The interplay among collagen concentration, particle size, and surface roughness was found to influence cell adhesion and proliferation. Adsorption and desorption tests showed the reversible attachment of the particles to magnetic sheets, enabling precise spatial control and targeted cell delivery. The particles demonstrated their utility as cell carriers, supporting cell migration and proliferation. These findings showcase the potential of CMPMBs as a promising platform for advanced cell delivery and tissue regeneration applications. The ability to fine-tune particle properties and manipulate them using magnetic fields offers new possibilities for creating complex tissue constructs and controlling cellular behavior, which could contribute to the development of more effective regenerative therapies and tissue engineering approaches.

Bioinspired Printable Tough Adhesives with In Situ Benignly Triggered Mechanical Enhancements

AbstractAdhesives can intimately connect humans to machines, seamlessly bond diverse tissues in the human body, and manage various diseases. However, the precise spatial control of wet and tough adhesion of biocompatible hydrogels on biological tissues remains a major challenge. Inspired by the bioglue secreted by sandcastle worms, the design of printable tough adhesives (PTAs) is proposed, a supramolecular hydrogel that can be printed into defined structures, in situ mechanically reinforced into a tough matrix with physiologically relevant benign triggers, and strongly adhere to diverse substrates. With carefully selected polymer components and ratios, it is discovered that the 3D printed PTAs can achieve a marked increase in toughness, tensile strength, and stiffness after being immersed in water/saline solution or attached to biological tissues. To assess the robust toughening mechanism triggered by the supramolecular interactions, the effects of polymer content and pH on the mechanical performance of PTAs and the kinetics of their triggered reinforcement are thoroughly investigated. The potential of PTAs is further demonstrated for manufacturing tough connective tissue mimetics, controlling patterned bioadhesion, and designing programmable 4D soft robotics. The bioinspired, printable, benignly triggerable, and adhesive supramolecular PTAs are expected to find broad applications in engineering and medicine.

3D printed carbon fiber reinforced carbon as an energy efficient alternative to graphite for EFAS tooling

As Electric Field Assisted Sintering (EFAS) gains more industrial acceptance and use, it becomes more important to develop more efficient means to implement this technology. To this aim, 3D printed continuous carbon fiber reinforced carbon (CCC) was manufactured and fabricated into tooling for EFAS systems as an alternative to traditional graphite tooling. The impact of fiber orientation on the thermal and electrical properties of the CCC was characterized. Sample material was sintered in Tokai G535 graphite tooling, under common processing conditions and compared with CCC tooling. There was nearly 50 % energy savings compared to graphite while maintaining equivalent sample density and microstructure plus keeping ram temperatures 39 % cooler. This is due to spatial control of generated heat and thermal diffusivity within the molds, by means of fiber orientation anisotropy. Finite element modeling of the tooling design supported the experimental results as well as displays the effect of optimization of this 3D printed CCC material.