Phase separation by low complexity domains promotes stress granule assembly and drives pathological fibrillization.

Stress granules are membrane-less organelles composed of RNA-binding proteins (RBPs) and RNA. Functional impairment of stress granules has been implicated in amyotrophic lateral sclerosis, frontotemporal dementia, and multisystem proteinopathy-diseases that are characterized by fibrillar inclusions of RBPs. Genetic evidence suggests a link between persistent stress granules and the accumulation of pathological inclusions. Here, we demonstrate that the disease-related RBP hnRNPA1 undergoes liquid-liquid phase separation (LLPS) into protein-rich droplets mediated by a low complexity sequence domain (LCD). While the LCD of hnRNPA1 is sufficient to mediate LLPS, the RNA recognition motifs contribute to LLPS in the presence of RNA, giving rise to several mechanisms for regulating assembly. Importantly, while not required for LLPS, fibrillization is enhanced in protein-rich droplets. We suggest that LCD-mediated LLPS contributes to the assembly of stress granules and their liquid properties and provides a mechanistic link between persistent stress granules and fibrillar protein pathology in disease.

Reduction of oligomer size modulates the competition between cluster formation and phase separation of the tumor suppressor SPOP.

Phase separation compartmentalizes many cellular pathways. Given that the same interactions that drive phase separation mediate the formation of soluble complexes below the saturation concentration, the contribution of condensates versus complexes to function is sometimes unclear. Here, we characterized several new cancer-associated mutations of the tumor suppressor speckle-type POZ protein (SPOP), a substrate recognition subunit of the Cullin3-RING ubiquitin ligase. This pointed to a strategy for generating separation-of-function mutations. SPOP self-associates into linear oligomers and interacts with multivalent substrates, and this mediates the formation of condensates. These condensates bear the hallmarks of enzymatic ubiquitination activity. We characterized the effect of mutations in the dimerization domains of SPOP on its linear oligomerization, binding to its substrate DAXX, and phase separation with DAXX. We showed that the mutations reduce SPOP oligomerization and shift the size distribution of SPOP oligomers to smaller sizes. The mutations therefore reduce the binding affinity to DAXX but unexpectedly enhance the poly-ubiquitination activity of SPOP toward DAXX. Enhanced activity may be explained by enhanced phase separation of DAXX with the SPOP mutants. Our results provide a comparative assessment of the functional role of complexes versus condensates and support a model in which phase separation is an important factor in SPOP function. Our findings also suggest that tuning of linear SPOP self-association could be used by the cell to modulate activity and provide insights into the mechanisms underlying hypermorphic SPOP mutations. The characteristics of cancer-associated SPOP mutations suggest a route for designing separation-of-function mutations in other phase-separating systems.

Reinforcement Learning-Based Joint Beamwidth and Beam Alignment Interval Optimization in V2I Communications.

The directional antenna combined with beamforming is one of the attractive solutions to accommodate high data rate applications in 5G vehicle communications. However, the directional nature of beamforming requires beam alignment between the transmitter and the receiver, which incurs significant signaling overhead. Hence, we need to find the optimal parameters for directional beamforming, i.e., the antenna beamwidth and beam alignment interval, that maximize the throughput, taking the beam alignment overhead into consideration. In this paper, we propose a reinforcement learning (RL)-based beamforming scheme in a vehicle-to-infrastructure system, where we jointly determine the antenna beamwidth and the beam alignment interval, taking into account the past and future rewards. The simulation results show that the proposed RL-based joint beamforming scheme outperforms conventional beamforming schemes in terms of the average throughput and the average link stability ratio.

Molecular features, antioxidant potential, and immunological expression assessment of thioredoxin-like protein 1 (TXNL1) in yellowtail clownfish (Amphiprion clarkii).

Thioredoxin-like protein 1 (TXNL1) is a redox-active protein belonging to the thioredoxin family, which mainly controls the redox status of cells. The TXNL1 gene from Amphiprion clarkii (AcTXNL1) was obtained from a pre-established transcriptome database. The AcTXNL1 is encoded with 289 amino acids and is predominantly localized in the cytoplasm and nucleus. The TXN domain of AcTXNL1 comprises a34CGPC37 motif with redox-reactive thiol (SH-) groups. The spatial distribution pattern of AcTXNL1 mRNA was examined in different tissues, and the muscle was identified as the highest expressed tissue. AcTXNL1 mRNA levels in the blood and gills were significantly increased in response to different immunostimulants. In vitro antioxidant capacity of the recombinant AcTXNL1 protein (rACTXNL1) was evaluated using the ABTS free radical-scavenging activity assay, cupric ion reducing antioxidant capacity assay, turbidimetric disulfide reduction assay, and DNA nicking protection assay. The potent antioxidant activity of rAcTXNL1 exhibited a concentration-dependent manner in all assays. Furthermore, in the cellular environment, overexpression of AcTXNL1 increased cell viability under H2O2 stress and reduced nitric oxide (NO) production induced by lipopolysaccharides (LPS). Collectively, the experimental results revealed that AcTXNL1 is an antioxidant and immunologically important gene in A. clarkii.

Aphid-Inspired and Thermally-Actuated Soft Gripper Using 3D Printing Technology.

Herein, a thermo-actuated aphid-inspired dry adhesive (TADA) that offers tunable and reversible adhesion is reported. It is easily fabricated through 3D printing using a polylactic acid (PLA) filament and silicone elastomer, avoiding the use of unfavorable methods for micro- and nanofabrication and unwanted particles for actuation. The tunable adhesive system mimics aphid biology to achieve adhesion switchability. Switching between adhesion states is enabled by the thermo-actuated PLA, which has shape memory properties. Additionally, silicone elastomer enables adherence to flat substrates such as glass, silicon wafers, and acrylic plates. The detachment time of the TADA can be controlled by changing the printing layer height, which is a 3D-printing parameter that results in a short detachment time when the printing layer height is small. The adhesion strength is measured by applying different preloads and varying the size of the adhesive area. The reversibility between the adhesion-on and adhesion-off states, revealing good repeatability with similar adhesion strengths is also demonstrated. The TADA has potential applications in transferring silicon wafers. In addition, it can be printed to fit a flat plate of any shape, enabling it to grip the plate stably.

Determination of Gas Permeation Properties in Polymer Using Capacitive Electrode Sensors.

The objective of this work was to develop an effective technique for characterizing the permeation properties of various gases, including H2, He, N2, and Ar, that are absorbed in polymers. Simultaneous three-channel real-time techniques for measuring the sorption content and diffusivity of gases emitted from polymers are developed after exposure to high pressure and the subsequent decompression of the corresponding gas. These techniques are based on the volumetric measurement of released gas combined with the capacitance measurement of the water content by both semi-cylindrical and coaxial-cylindrical electrodes. This minimizes the uncertainty due to the varying temperature and pressure of laboratory environments. The gas uptake and diffusivity are determined as a function of the exposed pressure and gas spices in nitrile butadiene rubber (NBR) and ethylene propylene diene monomer (EPDM) polymers. The pressure-dependent gas transport behaviors of four different gases are presented and compared with those obtained by different techniques. A linear correlation between the logarithmic diffusivity and kinetic diameter of molecules in the gas is found between the two polymers.

Higher-order oligomerization promotes localization of SPOP to liquid nuclear speckles.

Membrane-less organelles in cells are large, dynamic protein/protein or protein/RNA assemblies that have been reported in some cases to have liquid droplet properties. However, the molecular interactions underlying the recruitment of components are not well understood. Herein, we study how the ability to form higher-order assemblies influences the recruitment of the speckle-type POZ protein (SPOP) to nuclear speckles. SPOP, a cullin-3-RING ubiquitin ligase (CRL3) substrate adaptor, self-associates into higher-order oligomers; that is, the number of monomers in an oligomer is broadly distributed and can be large. While wild-type SPOP localizes to liquid nuclear speckles, self-association-deficient SPOP mutants have a diffuse distribution in the nucleus. SPOP oligomerizes through its BTB and BACK domains. We show that BTB-mediated SPOP dimers form linear oligomers via BACK domain dimerization, and we determine the concentration-dependent populations of the resulting oligomeric species. Higher-order oligomerization of SPOP stimulates CRL3(SPOP) ubiquitination efficiency for its physiological substrate Gli3, suggesting that nuclear speckles are hotspots of ubiquitination. Dynamic, higher-order protein self-association may be a general mechanism to concentrate functional components in membrane-less cellular bodies.

Simplified protein design biased for prebiotic amino acids yields a foldable, halophilic protein.

A compendium of different types of abiotic chemical syntheses identifies a consensus set of 10 "prebiotic" α-amino acids. Before the emergence of biosynthetic pathways, this set is the most plausible resource for protein formation (i.e., proteogenesis) within the overall process of abiogenesis. An essential unsolved question regarding this prebiotic set is whether it defines a "foldable set"--that is, does it contain sufficient chemical information to permit cooperatively folding polypeptides? If so, what (if any) characteristic properties might such polypeptides exhibit? To investigate these questions, two "primitive" versions of an extant protein fold (the ß-trefoil) were produced by top-down symmetric deconstruction, resulting in a reduced alphabet size of 12 or 13 amino acids and a percentage of prebiotic amino acids approaching 80%. These proteins show a substantial acidification of pI and require high salt concentrations for cooperative folding. The results suggest that the prebiotic amino acids do comprise a foldable set within the halophile environment.

Experimental support for the evolution of symmetric protein architecture from a simple peptide motif.

The majority of protein architectures exhibit elements of structural symmetry, and "gene duplication and fusion" is the evolutionary mechanism generally hypothesized to be responsible for their emergence from simple peptide motifs. Despite the central importance of the gene duplication and fusion hypothesis, experimental support for a plausible evolutionary pathway for a specific protein architecture has yet to be effectively demonstrated. To address this question, a unique "top-down symmetric deconstruction" strategy was utilized to successfully identify a simple peptide motif capable of recapitulating, via gene duplication and fusion processes, a symmetric protein architecture (the threefold symmetric ß-trefoil fold). The folding properties of intermediary forms in this deconstruction agree precisely with a previously proposed "conserved architecture" model for symmetric protein evolution. Furthermore, a route through foldable sequence-space between the simple peptide motif and extant protein fold is demonstrated. These results provide compelling experimental support for a plausible evolutionary pathway of symmetric protein architecture via gene duplication and fusion processes.

An asynchronous wireless network for capturing event-driven data from large populations of autonomous sensors.

Networks of spatially distributed radiofrequency identification sensors could be used to collect data in wearable or implantable biomedical applications. However, the development of scalable networks remains challenging. Here we report a wireless radiofrequency network approach that can capture sparse event-driven data from large populations of spatially distributed autonomous microsensors. We use a spectrally efficient, low-error-rate asynchronous networking concept based on a code-division multiple-access method. We experimentally demonstrate the network performance of several dozen submillimetre-sized silicon microchips and complement this with large-scale in silico simulations. To test the notion that spike-based wireless communication can be matched with downstream sensor population analysis by neuromorphic computing techniques, we use a spiking neural network machine learning model to decode prerecorded open source data from eight thousand spiking neurons in the primate cortex for accurate prediction of hand movement in a cursor control task.

The Synergistic Effect of Carbon Black/Carbon Nanotube Hybrid Fillers on the Physical and Mechanical Properties of EPDM Composites after Exposure to High-Pressure Hydrogen Gas.

This study investigated the synergistic effect of carbon black/multi-wall carbon nanotube (CB/MWCNT) hybrid fillers on the physical and mechanical properties of Ethylene propylene diene rubber (EPDM) composites after exposure to high-pressure hydrogen gas. The EPDM/CB/CNT hybrid composites were prepared by using the EPDM/MWCNT master batch (MB) with 10 phr CNTs to enhance the dispersion of CNTs in hybrid composites. The investigation included a detailed analysis of cure characteristics, crosslink density, Payne effect, mechanical properties, and hydrogen permeation properties. After exposure to 96.3 MPa hydrogen gas, the hydrogen uptake and the change in volume and mechanical properties of the composites were assessed. We found that as the MWCNT volume fraction in fillers increased, the crosslink density, filler-filler interaction, and modulus of hybrid composites increased. The hydrogen uptake and the solubility of the composites decreased with an increasing MWCNT volume fraction in fillers. Moreover, after exposure to hydrogen gas, the change in volume and mechanical properties exhibited a diminishing trend with a higher MWCNT volume fraction. We conclude that the hybridization of CB and CNTs formed strong filler-filler networks in hybrid composites, consequently reinforcing the EPDM composites and enhancing the barrier properties of hydrogen gas.

Molecular characterization, cytoprotective, DNA protective, and immunological assessment of peroxiredoxin-1 (Prdx1) from yellowtail clownfish (Amphiprion clarkii).

Peroxiredoxin-1 (Prdx1) is a thiol-specific antioxidant enzyme that detoxifies reactive oxygen species (ROS) and regulates the redox status of cells. In this study, the Prdx1 cDNA sequence was isolated from the pre-established Amphiprion clarkii (A. clarkii) (AcPrdx1) transcriptome database and characterized structurally and functionally. The AcPrdx1 coding sequence comprises 597 bp and encodes 198 amino acids with a molecular weight of 22.1 kDa and a predicted theoretical isoelectric point of 6.3. AcPrdx1 is localized and functionally available in the cytoplasm and nucleus of cells. The TXN domain of AcPrdx1 comprises two peroxiredoxin signature VCP motifs, which contain catalytic peroxidatic (Cp-C52) and resolving cysteine (CR-C173) residues. The constructed phylogenetic tree and sequence alignment revealed that AcPrdx1 is evolutionarily conserved, and its most closely related counterpart is Amphiprion ocellaris. Under normal physiological conditions, AcPrdx1 was ubiquitously detected in all tissues examined, with the most robust expression in the spleen. Furthermore, AcPrdx1 transcripts were significantly upregulated in the spleen, head kidney, and blood after immune stimulation by polyinosinic:polycytidylic acid (poly (I:C)), lipopolysaccharide (LPS), and Vibrio harveyi injection. Recombinant AcPrdx1 (rAcPrdx1) demonstrated antioxidant and DNA protective properties in a concentration-dependent manner, as evidenced by insulin disulfide reduction, peroxidase activity, and metal-catalyzed oxidation (MCO) assays, whereas cells transfected with pcDNA3.1(+)/AcPrdx1 showed significant cytoprotective function under oxidative and nitrosative stress. Overexpression of AcPrdx1 in fathead minnow (FHM) cells led to a lower viral copy number following viral hemorrhagic septicemia virus (VHSV) infection, along with upregulation of several antiviral genes. Collectively, this study provides insights into the function of AcPrdx1 in defense against oxidative stressors and its role in the immune response against pathogenic infections in A. clarkii.

Tunable porous organic crystals: structural scope and adsorption properties of nanoporous steroidal ureas.

Previous work has shown that certain steroidal bis-(N-phenyl)ureas, derived from cholic acid, form crystals in the P6(1) space group with unusually wide unidimensional pores. A key feature of the nanoporous steroidal urea (NPSU) structure is that groups at either end of the steroid are directed into the channels and may in principle be altered without disturbing the crystal packing. Herein we report an expanded study of this system, which increases the structural variety of NPSUs and also examines their inclusion properties. Nineteen new NPSU crystal structures are described, to add to the six which were previously reported. The materials show wide variations in channel size, shape, and chemical nature. Minimum pore diameters vary from ~0 up to 13.1 Å, while some of the interior surfaces are markedly corrugated. Several variants possess functional groups positioned in the channels with potential to interact with guest molecules. Inclusion studies were performed using a relatively accessible tris-(N-phenyl)urea. Solvent removal was possible without crystal degradation, and gas adsorption could be demonstrated. Organic molecules ranging from simple aromatics (e.g., aniline and chlorobenzene) to the much larger squalene (M(w) = 411) could be adsorbed from the liquid state, while several dyes were taken up from solutions in ether. Some dyes gave dichroic complexes, implying alignment of the chromophores in the NPSU channels. Notably, these complexes were formed by direct adsorption rather than cocrystallization, emphasizing the unusually robust nature of these organic molecular hosts.

Emergence of symmetric protein architecture from a simple peptide motif: evolutionary models.

Structural symmetry is observed in the majority of fundamental protein folds and gene duplication and fusion evolutionary processes are postulated to be responsible. However, convergent evolution leading to structural symmetry has also been proposed; additionally, there is debate regarding the extent to which exact primary structure symmetry is compatible with efficient protein folding. Issues of symmetry in protein evolution directly impact strategies for de novo protein design as symmetry can substantially simplify the design process. Additionally, when considering gene duplication and fusion in protein evolution, there are two competing models: "emergent architecture" and "conserved architecture". Recent experimental work has shed light on both the evolutionary process leading to symmetric protein folds as well as the ability of symmetric primary structure to efficiently fold. Such studies largely support a "conserved architecture" evolutionary model, suggesting that complex protein architecture was an early evolutionary achievement involving oligomerization of smaller polypeptides.

The effect of Er:YAG laser irradiation on hydroxyapatite-coated implants and fluoride-modified TiO2-blasted implant surfaces: a microstructural analysis.

The purpose of this study was to evaluate the microscopic changes and surface roughness on hydroxyapatite (HA)-coated implants following exposure to different powers and durations of Er:YAG laser irradiation in order to determine the proper pulse energy level and irradiation time. Ten HA-coated implants and ten fluoride-modified TiO2 implants were used. The implants were divided into a control (one implant) and test group (nine implants) for each implant type. Implants in the test groups were sub-divided into three groups (three implants per group) based on the applied laser pulse energy and irradiation time. The measurement of surface roughness was performed on all implants in the test groups using a white light interferometer before and after laser irradiation. R a values were recorded and compared in order to evaluate changes in surface roughness. For HA-coated implants, the R a values increased in all test groups after laser irradiation. However, mean R a values in the fluoride-modified TiO2-blasted implant test group were decreased after irradiation. There was no statistical difference. Scanning electron microscope analysis revealed surface alterations in both the HA-coated and fluoridated TiO2-blasted implants irradiated for 1.5 min at 100 mJ/pulse, 10 Hz. When the pulse energy and irradiation time increased, greater surface alterations, including surface flattening and microfractures, were observed. In conclusion, the results of the current study suggest that no changes could be observed in both HA-coated implants and fluoride-modified TiO2-blasted implants after irradiation at an intensity of 100 mJ/pulse, 10 Hz for 1 min performed to achieve surface detoxification.

3D-printing-assisted flexible pressure sensor with a concentric circle pattern and high sensitivity for health monitoring.

In this study, a flexible pressure sensor is fabricated using polydimethylsiloxane (PDMS) with a concentric circle pattern (CCP) obtained through a fused deposition modeling (FDM)-type three-dimensional (3D) printer and poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) as the active layer. Through layer-by-layer additive manufacturing, the CCP surface is generated from a thin cone model with a rough surface by the FDM-type 3D printer. A novel compression method is employed to convert the cone shape into a planar microstructure above the glass transition temperature of a polylactic acid (PLA) filament. To endow the CCP surface with conductivity, PDMS is used to replicate the compressed PLA, and PEDOT:PSS is coated by drop-casting. The size of the CCP is controlled by changing the printing layer height (PLH), which is one of the 3D printing parameters. The sensitivity increases as the PLH increases, and the pressure sensor with a 0.16 mm PLH exhibits outstanding sensitivity (160 kPa-1), corresponding to a linear pressure range of 0-0.577 kPa with a good linearity of R 2 = 0.978, compared to other PLHs. This pressure sensor exhibits stable and repeatable operation under various pressures and durability under 6.56 kPa for 4000 cycles. Finally, monitoring of various health signals such as those for the wrist pulse, swallowing, and pronunciation of words is demonstrated as an application. These results support the simple fabrication of a highly sensitive, flexible pressure sensor for human health monitoring.

Versatile On-Chip Programming of Circuit Hardware for Wearable and Implantable Biomedical Microdevices.

Wearable and implantable microscale electronic sensors have been developed for a range of biomedical applications. The sensors, typically millimeter size silicon microchips, are sought for multiple sensing functions but are severely constrained by size and power. To address these challenges, a hardware programmable application-specific integrated circuit design is proposed and post-process methodology is exemplified by the design of battery-less wireless microchips. Specifically, both mixed-signal and radio frequency circuits are designed by incorporating metal fuses and anti-fuses on the top metal layer to enable programmability of any number of features in hardware of the system-on-chip (SoC) designs. This is accomplished in post-foundry editing by combining laser ablation and focused ion beam processing. The programmability provided by the technique can significantly accelerate the SoC chip development process by enabling the exploration of multiple internal circuit parameters without the requirement of additional programming pads or extra power consumption. As examples, experimental results are described for sub-millimeter size complementary metal-oxide-semiconductor microchips being developed for wireless electroencephalogram sensors and as implantable microstimulators for neural interfaces. The editing technique can be broadly applicable for miniaturized biomedical wearables and implants, opening up new possibilities for their expedited development and adoption in the field of smart healthcare.

Investigation of the Gas Permeation Properties Using the Volumetric Analysis Technique for Polyethylene Materials Enriched with Pure Gases under High Pressure: H2, He, N2, O2 and Ar.

Polyethylene (PE) is widely used as a gas-sealing material in packing films and gas transport pipes. A technique for evaluating the permeability of water-insoluble gases has recently been developed. This technique is a volumetric analysis that is used to calculate the gas permeability by measuring the gas uptake and diffusivity. With this technique, we investigated the permeability of pure gases, such as H2, He, N2, O2 and Ar, enriched under high pressure up to 9 MPa in low-density polyethylene (LDPE), ultrahigh molecular weight polyethylene (UHMWPE) and high-density polyethylene (HDPE). The gas uptake showed a linear pressure-dependent behavior that followed Henry's law, and the diffusivity was independent of the pressure. Furthermore, the logarithmic diffusivity values of the five gases linearly decreased as their molecular kinetic diameters increased. The logarithmic solubility values linearly increased as the critical temperatures of the gases increased. The calculated permeability results were correlated with the volume fraction of the amorphous phase and the fractional free volume. This result newly showed that the amorphous phase was directly correlated to the fractional free volume.

Metaverse Wearables for Immersive Digital Healthcare: A Review.

The recent exponential growth of metaverse technology has been instrumental in reshaping a myriad of sectors, not least digital healthcare. This comprehensive review critically examines the landscape and future applications of metaverse wearables toward immersive digital healthcare. The key technologies and advancements that have spearheaded the metamorphosis of metaverse wearables are categorized, encapsulating all-encompassed extended reality, such as virtual reality, augmented reality, mixed reality, and other haptic feedback systems. Moreover, the fundamentals of their deployment in assistive healthcare (especially for rehabilitation), medical and nursing education, and remote patient management and treatment are investigated. The potential benefits of integrating metaverse wearables into healthcare paradigms are multifold, encompassing improved patient prognosis, enhanced accessibility to high-quality care, and high standards of practitioner instruction. Nevertheless, these technologies are not without their inherent challenges and untapped opportunities, which span privacy protection, data safeguarding, and innovation in artificial intelligence. In summary, future research trajectories and potential advancements to circumvent these hurdles are also discussed, further augmenting the incorporation of metaverse wearables within healthcare infrastructures in the post-pandemic era.

One-Step Synthesis of Transition Metal Dichalcogenide Quantum Dots Using Only Alcohol Solvents for Indoor-Light Photocatalytic Antibacterial Activity.

In this study, we report a one-step direct synthesis of molybdenum disulfide (MoS2) and tungsten disulfide (WS2) quantum dots (QDs) through a solvothermal reaction using only alcohol solvents and efficient Escherichia coli (E. coli) decompositions as photocatalytic antibacterial agents under visible light irradiation. The solvothermal reaction gives the scission of molybdenum-sulfur (Mo-S) and tungsten-sulfur (W-S) bonding during the synthesis of MoS2 and WS2 QDs. Using only alcohol solvent does not require a residue purification process necessary for metal intercalation. As the number of the CH3 groups of alcohol solvents among ethyl, isopropyl, and tert(t)-butyl alcohols increases, the dispersibility of MoS2/WS2 increases. The CH3 groups of alcohols minimize the surface energy, leading to the effective exfoliation and disintegration of the bulk under heat and pressure. The bulky t-butyl alcohol with the highest number of methyl groups shows the highest exfoliation and yield. MoS2 QDs with a lateral size of about 2.5 nm and WS2 QDs of about 10 nm are prepared, exhibiting a strong blue luminescence under 365 nm ultraviolet (UV) light irradiation. Their heights are 0.68-3 and 0.72-5 nm, corresponding to a few layers of MoS2 and WS2, respectively. They offer a highly efficient performance in sterilizing E. coli as the visible-light-driven photocatalyst.