Eco-Friendly Textile-Based Wearable Humidity Sensor with Multinode Wireless Connectivity for Healthcare Applications.

Textile-based wearable humidity sensors are of great interest for human healthcare monitoring as they can provide critical human-physiology information. The demand for wearable and sustainable sensing technology has significantly promoted the development of eco-friendly sensing solutions for potential real-world applications. Herein, a biodegradable cotton (textile)-based wearable humidity sensor has been developed using fabsil-treated cotton fabric coated with a poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) sensing layer. The structural, chemical composition, hygroscopicity, and morphological properties are examined using X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), contact angle measurement, and scanning electron microscopy (SEM) analysis. The developed sensor exhibited a nearly linear response (Adj. R-square value observed as 0.95035) over a broad relative humidity (RH) range from 25 to 91.5%RH displaying high sensitivity (26.1%/%RH). The sensor shows excellent reproducibility (on replica sensors with a margin of error ±1.98%) and appreciable stability/aging with time (>4.5 months), high flexibility (studied at bending angles 30°, 70°, 120°, and 150°), substantial response/recovery durations (suitable for multiple applications), and highly repeatable (multicyclic analysis) sensing performance. The prospective relevance of the developed humidity sensor toward healthcare applications is demonstrated via breathing rate monitoring (via a sensor attached to a face mask), distinguishing different breathing patterns (normal, deep, and fast), skin moisture monitoring, and neonatal care (diaper wetting). The multinode wireless connectivity is demonstrated using a Raspberry Pi Pico-based system for demonstrating the potential applicability of the developed sensor as a real-time humidity monitoring system for the healthcare sector. Further, the biodegradability analysis of the used textile is evaluated using the soil burial degradation test. The work suggests the potential applicability of the developed flexible and eco-friendly humidity sensor in wearable healthcare devices and other humidity sensing applications.

Toward quantitative super-resolution methods for cryo-CLEM.

Cryogenic ultrastructural imaging techniques such as cryo-electron tomography have produced a revolution in how the structure of biological systems is investigated by enabling the determination of structures of protein complexes immersed in a complex biological matrix within vitrified cell and model organisms. However, so far, the portfolio of successes has been mostly limited to highly abundant complexes or to structures that are relatively unambiguous and easy to identify through electron microscopy. In order to realize the full potential of this revolution, researchers would have to be able to pinpoint lower abundance species and obtain functional annotations on the state of objects of interest which would then be correlated to ultrastructural information to build a complete picture of the structure-function relationships underpinning biological processes. Fluorescence imaging at cryogenic conditions has the potential to be able to meet these demands. However, wide-field images acquired at low numeric aperture (NA) using air immersion objective have a low resolving power and cannot provide accurate enough three-dimensional (3D) localization to enable the assignment of functional annotations to individual objects of interest or target sample debulking to ensure the preservation of the structures of interest. It is therefore necessary to develop super-resolved cryo-fluorescence workflows capable of fulfilling this role and enabling new biological discoveries. In this chapter, we present the current state of development of two super-resolution cryogenic fluorescence techniques, superSIL-STORM and astigmatism-based 3D STORM, show their application to a variety of biological systems and discuss their advantages and limitations. We further discuss the future applicability to cryo-CLEM workflows though examples of practical application to the study of membrane protein complexes both in mammalian cells and in Escherichia coli.

The two C. elegans class VI myosins, SPE-15/HUM-3 and HUM-8, share similar motor properties, but have distinct developmental and tissue expression patterns.

Myosins of class VI move toward the minus-end of actin filaments and play vital roles in cellular processes such as endocytosis, autophagy, protein secretion, and the regulation of actin filament dynamics. In contrast to the majority of metazoan organisms examined to date which contain a single MYO6 gene, C. elegans, possesses two MYO6 homologues, SPE-15/HUM-3 and HUM-8. Through a combination of in vitro biochemical/biophysical analysis and cellular assays, we confirmed that both SPE-15/HUM-3 and HUM-8 exhibit reverse directionality, velocities, and ATPase activity similar to human MYO6. Our characterization also revealed that unlike SPE-15/HUM-3, HUM-8 is expressed as two distinct splice isoforms, one with an additional unique 14 amino acid insert in the cargo-binding domain. While lipid and adaptor binding sites are conserved in SPE-15/HUM-3 and HUM-8, this conservation does not enable recruitment to endosomes in mammalian cells. Finally, we performed super-resolution confocal imaging on transgenic worms expressing either mNeonGreen SPE-15/HUM-3 or wrmScarlet HUM-8. Our results show a clear distinction in tissue distribution between SPE-15/HUM-3 and HUM-8. While SPE-15/HUM-3 exhibited specific expression in the gonads and neuronal tissue in the head, HUM-8 was exclusively localized in the intestinal epithelium. Overall, these findings align with the established tissue distributions and localizations of human MYO6.

The Effect of Chitosan Incorporation on Physico-Mechanical and Biological Characteristics of a Calcium Silicate Filling Material.

OBJECTIVES: A tricalcium silicate-based cement, Biodentine™, has displayed antibiofilm activity when mixed with chitosan powder. This study aimed to assess the effect of chitosan incorporation on the physico-mechanical and biological properties of Biodentine™. METHODS: In this study, medium molecular weight chitosan powder was incorporated into Biodentine™ in varying proportions (2.5 wt%, 5 wt%, 10 wt%, and 20 wt%). The setting time was determined using a Vicat apparatus, solubility was assessed by calculating weight variation after water immersion, radiopacity was evaluated and expressed in millimeters of aluminum, the compressive strength was evaluated using an Instron testing machine, and the microhardness was measured with a Vickers microhardness tester. In addition, surface topography of specimens was analyzed using scanning electron microscopy, and the effect of chitosan on the viability of human embryonic kidney (HEK 293) cells was measured by a colorimetric MTT assay. RESULTS: Incorporation of 2.5 wt% and 5 wt% chitosan powder delivered an advantage by speeding up the setting time of Biodentine material. However, the incorporation of chitosan compromised all other material properties and the crystalline structure in a dose-dependent manner. The chitosan-modified material also showed significant decreases in the proliferation of the HEK 293 cells, signifying decreased biocompatibility. SIGNIFICANCE: Chitosan incorporation into calcium silicate materials adversely affects the physical and biological properties of the material. Despite the increased antimicrobial activity of the modified material, the diminution in these properties is likely to reduce its clinical value.

A mechanistic study identifying improved technology critical metal delamination from printed circuit boards at lower power sonications in a deep eutectic solvent.

Deep eutectic solvents (DESs) are an emerging class of ionic liquids that offer a solution to reclaiming technology critical metals (TCMs) from electronic waste, with potential for improved life cycle analysis. The high viscosities typical of DESs, however, impose mass transport limitations such that passive TCM removal generally requires immersion over extended durations, in some cases in the order of hours. It is postulated that, through the targeted application of power ultrasound, delamination of key structures in electronic components immersed in DESs can be significantly accelerated, thereby enabling rapid recovery of TCMs. In this paper, we fully characterise cavitation in a Choline Chloride-Ethylene Glycol DES as a function of sonotrode input power, by the acoustic detection of the bubble collapse shockwave content generated during sonications at more than 20 input powers over the available range. This justifies the selection of two powers for a detailed study of ultrasonically enhanced TCM-delamination from printed circuit boards (PCBs). Dual-perspective high-speed imaging is employed, which facilitates simultaneous observation of TCM removal, and the cavitation evolution and interaction with the PCB surface. Bubble jetting is identified as a key contributor to initial pitting of the TCM layers, exposing the larger underlying copper layer, with the contributions of additional inertial cavitation-mediated phenomena such as bubble-collapse shockwaves also demonstrated as important for delamination. Optimal cavitation activity throughout the sonication then promotes etching of the copper base layer of the PCB structure targeted by the DES, liberating the overlaying TCMs in sections as large as 0.79 mm2. We report a thirtyfold improvement in processing time compared to passive delamination, with sonications at the lower power outperforming those at the higher power. The results demonstrate the potential for industrially scalable recovery of TCMs from the growing quantities of global e-waste, using combined power ultrasonics and DESs.

Optimized Production and Analysis of Recombinant Protein-Filled Vesicles from E. coli.

This innovative system, using a short peptide tag, that exports multiple recombinant proteins in membrane bound vesicles from E. coli, provides an effective solution to a range of problems associated with bacterial recombinant protein expression. These recombinant vesicles compartmentalise proteins within a micro-environment that facilitates the production of otherwise challenging, toxic, insoluble, or disulfide-bond containing proteins from bacteria. Protein yield is increased considerably when compared to typical bacterial expression in the absence of the vesicle-nucleating peptide tag. The release of vesicle-packaged proteins supports isolation from the culture medium and permits long-term active protein storage. This technology gives rise to increased yields of vesicle-packaged, functional proteins for simplified downstream processing for a diverse range of applications from applied biotechnology to discovery science and medicine. In the present article and the associated video, a detailed protocol of the method is provided, which highlights key steps in the methodology to maximize recombinant protein-filled vesicle production.

Conductive polymer based hydrogels and their application in wearable sensors: a review.

Hydrogels have been attracting increasing attention for application in wearable electronics, due to their intrinsic biomimetic features, highly tunable chemical-physical properties (mechanical, electrical, etc.), and excellent biocompatibility. Among many proposed varieties of hydrogels, conductive polymer-based hydrogels (CPHs) have emerged as a promising candidate for future wearable sensor designs, with capability of realizing desired features using different tuning strategies ranging from molecular design (with a low length scale of 10-10 m) to a micro-structural configuration (up to a length scale of 10-2 m). However, considerable challenges remain to be overcome, such as the limited strain sensing range due to the mechanical strength, the signal loss/instability caused by swelling/deswelling, the significant hysteresis of sensing signals, the de-hydration induced malfunctions, and the surface/interfacial failure during manufacturing/processing. This review aims to offer a targeted scan of recent advancements in CPH based wearable sensor technology, from the establishment of dedicated structure-property relationships in the lab to the advanced manufacturing routes for potential scale-up production. The application of CPHs in wearable sensors is also explored, with suggested new research avenues and prospects for CPHs in the future also included.

Controlling the structure of supramolecular fibre formation for benzothiazole based hydrogels with antimicrobial activity against methicillin resistant Staphylococcus aureus.

Antimicrobial resistance is one of the greatest threats to human health. Gram-positive methicillin resistant Staphylococcus aureus (MRSA), in both its planktonic and biofilm form, is of particular concern. Herein we identify the hydrogelation properties for a series of intrinsically fluorescent, structurally related supramolecular self-associating amphiphiles and determine their efficacy against both planktonic and biofilm forms of MRSA. To further explore the potential translation of this hydrogel technology for real-world applications, the toxicity of the amphiphiles was determined against the eukaryotic multicellular model organism, Caenorhabditis elegans. Due to the intrinsic fluorescent nature of these supramolecular amphiphiles, material characterisation of their molecular self-associating properties included; comparative optical density plate reader assays, rheometry and widefield fluorescence microscopy. This enabled determination of amphiphile structure and hydrogel sol dependence on resultant fibre formation.

High-yield vesicle-packaged recombinant protein production from E. coli.

We describe an innovative system that exports diverse recombinant proteins in membrane-bound vesicles from E. coli. These recombinant vesicles compartmentalize proteins within a micro-environment that enables production of otherwise challenging insoluble, toxic, or disulfide-bond containing proteins from bacteria. The release of vesicle-packaged proteins supports isolation from the culture and allows long-term storage of active protein. This technology results in high yields of vesicle-packaged, functional proteins for efficient downstream processing for a wide range of applications from discovery science to applied biotechnology and medicine.

Fluorescence and phosphorescence lifetime imaging reveals a significant cell nuclear viscosity and refractive index changes upon DNA damage.

Cytoplasmic viscosity is a crucial parameter in determining rates of diffusion-limited reactions. Changes in viscosity are associated with several diseases, whilst nuclear viscosity determines gene integrity, regulation and expression. Yet how drugs including DNA-damaging agents affect viscosity is unknown. We demonstrate the use of a platinum complex, Pt[L]Cl, that localizes efficiently mostly in the nucleus as a probe for nuclear viscosity. The phosphorescence lifetime of Pt[L]Cl is sensitive to viscosity and provides an excellent tool to investigate the impact of DNA damage. We show using Fluorescence Lifetime Imaging (FLIM) that the lifetime of both green and red fluorescent proteins (FP) are also sensitive to changes in cellular viscosity and refractive index. However, Pt[L]Cl proved to be a more sensitive viscosity probe, by virtue of microsecond phosphorescence lifetime versus nanosecond fluorescence lifetime of FP, hence greater sensitivity to bimolecular reactions. DNA damage was inflicted by either a two-photon excitation, one-photon excitation microbeam and X-rays. DNA damage of live cells causes significant increase in the lifetime of either Pt[L]Cl (HeLa cells, 12.5-14.1 µs) or intracellularly expressed mCherry (HEK293 cells, 1.54-1.67 ns), but a decrease in fluorescence lifetime of GFP from 2.65 to 2.29 ns (in V15B cells). These values represent a viscosity change from 8.59 to 20.56 cP as well as significant changes in the refractive index (RI), according to independent calibration. Interestingly DNA damage localized to a submicron region following a laser microbeam induction showed a whole cell viscosity change, with those in the nucleus being greater than the cytoplasm. We also found evidence of a by-stander effect, whereby adjacent un-irradiated cells also showed nuclear viscosity change. Finally, an increase in viscosity following DNA damage was also observed in bacterial cells with an over-expressed mNeonGreen FP, evidenced by the change in its lifetime from 2.8 to 2.4 ns.

High-Performance Triboelectric Nanogenerators Based on Commercial Textiles: Electrospun Nylon 66 Nanofibers on Silk and PVDF on Polyester.

A high-performance textile triboelectric nanogenerator is developed based on the common commercial fabrics silk and polyester (PET). Electrospun nylon 66 nanofibers were used to boost the tribo-positive performance of silk, and a poly(vinylidene difluoride) (PVDF) coating was deployed to increase the tribo-negativity of PET. The modifications confer a very significant boost in performance: output voltage and short-circuit current density increased ∼17 times (5.85 to 100 V) and ∼16 times (1.6 to 24.5 mA/m2), respectively, compared with the Silk/PET baseline. The maximum power density was 280 mW/m2 at a 4 MΩ resistance. The performance boost likely results from enhancing the tribo-positivity (and tribo-negativity) of the contact layers and from increased contact area facilitated by the electrospun nanofibers. Excellent stability and durability were demonstrated: the nylon nanofibers and PVDF coating provide high output, while the silk and PET substrate fabrics confer strength and flexibility. Rapid capacitor charging rates of 0.045 V/s (2 µF), 0.031 V/s (10 µF), and 0.011 V/s (22 µF) were demonstrated. Advantages include high output, a fully textile structure with excellent flexibility, and construction based on cost-effective commercial fabrics. The device is ideal as a power source for wearable electronic devices, and the approach can easily be deployed for other textiles.

Distinct actin-tropomyosin cofilament populations drive the functional diversification of cytoskeletal myosin motor complexes.

The effects of N-terminal acetylation of the high molecular weight tropomyosin isoforms Tpm1.6 and Tpm2.1 and the low molecular weight isoforms Tpm1.12, Tpm3.1, and Tpm4.2 on the actin affinity and the thermal stability of actin-tropomyosin cofilaments are described. Furthermore, we show how the exchange of cytoskeletal tropomyosin isoforms and their N-terminal acetylation affects the kinetic and chemomechanical properties of cytoskeletal actin-tropomyosin-myosin complexes. Our results reveal the extent to which the different actin-tropomyosin-myosin complexes differ in their kinetic and functional properties. The maximum sliding velocity of the actin filament as well as the optimal motor density for continuous unidirectional movement, parameters that were previously considered to be unique and invariant properties of each myosin isoform, are shown to be influenced by the exchange of the tropomyosin isoform and the N-terminal acetylation of tropomyosin.

Electrode materials for stretchable triboelectric nanogenerator in wearable electronics.

Stretchable Triboelectric Nanogenerators (TENGs) for wearable electronics are in significant demand in the area of self-powered energy harvesting and storage devices. Designing a suitable electrode is one of the major challenges in developing a fully wearable TENG device and requires research aimed at exploring new materials and methods to develop stretchable electrodes. This review article is dedicated to presenting recent developments in exploring new materials for flexible TENGs with special emphasis on electrode components for wearable devices. In addition, materials that can potentially deliver properties such as transparency, self-healability and water-resistance are also reviewed. Inherently stretchable materials and a combination of soft and rigid materials including polymers and their composites, inorganic and ceramic materials, 2D materials and carbonaceous nanomaterials are also addressed. Additionally, various fabrication strategies and geometrical patterning techniques employed for designing highly stretchable electrodes for wearable TENG devices are also explored. The challenges reflected in the present approaches as well as feasible suggestions for future advancements are discussed.

Acetylation stabilises calmodulin-regulated calcium signalling.

Calmodulin is a conserved calcium signalling protein that regulates a wide range of cellular functions. Amino-terminal acetylation is a ubiquitous post-translational modification that affects the majority of human proteins, to stabilise structure, as well as regulate function and proteolytic degradation. Here, we present data on the impact of amino-terminal acetylation upon structure and calcium signalling function of fission yeast calmodulin. We show that NatA-dependent acetylation stabilises the helical structure of the Schizosaccharomyces pombe calmodulin, impacting its ability to associate with myosin at endocytic foci. We go on to show that this conserved modification impacts both the calcium-binding capacity of yeast and human calmodulins. These findings have significant implications for research undertaken into this highly conserved essential protein.

Di-anionic self-associating supramolecular amphiphiles (SSAs) as antimicrobial agents against MRSA and Escherichia coli.

Herein, we report a series of di-anionic supramolecular self-associating amphiphiles (SSAs). We elucidate the antimicrobial properties of these SSAs against both methicillin resistant Staphylococcus aureus and Escherichia coli. In addition, we show this class of compound to form both intra- and intermolecular hydrogen bonded macrocyclic structures in the solid state.

Identification of sequence changes in myosin II that adjust muscle contraction velocity.

The speed of muscle contraction is related to body size; muscles in larger species contract at slower rates. Since contraction speed is a property of the myosin isoform expressed in a muscle, we investigated how sequence changes in a range of muscle myosin II isoforms enable this slower rate of muscle contraction. We considered 798 sequences from 13 mammalian myosin II isoforms to identify any adaptation to increasing body mass. We identified a correlation between body mass and sequence divergence for the motor domain of the 4 major adult myosin II isoforms (ß/Type I, IIa, IIb, and IIx), suggesting that these isoforms have adapted to increasing body mass. In contrast, the non-muscle and developmental isoforms show no correlation of sequence divergence with body mass. Analysis of the motor domain sequence of ß-myosin (predominant myosin in Type I/slow and cardiac muscle) from 67 mammals from 2 distinct clades identifies 16 sites, out of 800, associated with body mass (padj < 0.05) but not with the clade (padj > 0.05). Both clades change the same small set of amino acids, in the same order from small to large mammals, suggesting a limited number of ways in which contraction velocity can be successfully manipulated. To test this relationship, the 9 sites that differ between human and rat were mutated in the human ß-myosin to match the rat sequence. Biochemical analysis revealed that the rat-human ß-myosin chimera functioned like the native rat myosin with a 2-fold increase in both motility and in the rate of ADP release from the actin-myosin crossbridge (the step that limits contraction velocity). Thus, these sequence changes indicate adaptation of ß-myosin as species mass increased to enable a reduced contraction velocity and heart rate.

Identification of organophosphorus simulants for the development of next-generation detection technologies.

Organophosphorus (OP) chemical warfare agents (CWAs) represent an ongoing threat but the understandable widespread prohibition of their use places limitations on the development of technologies to counter the effects of any OP CWA release. Herein, we describe new, accessible methods for the identification of appropriate molecular simulants to mimic the hydrogen bond accepting capacity of the P[double bond, length as m-dash]O moiety, common to every member of this class of CWAs. Using the predictive methodologies developed herein, we have identified OP CWA hydrogen bond acceptor simulants for soman and sarin. It is hoped that the effective use of these physical property specific simulants will aid future countermeasure developments.

Yeasts as Complementary Model Systems for the Study of the Pathological Repercussions of Enhanced Synphilin-1 Glycation and Oxidation.

Synphilin-1 has previously been identified as an interaction partner of α-Synuclein (αSyn), a primary constituent of neurodegenerative disease-linked Lewy bodies. In this study, the repercussions of a disrupted glyoxalase system and aldose reductase function on Synphilin-1 inclusion formation characteristics and cell growth were investigated. To this end, either fluorescent dsRed-tagged or non-tagged human SNCAIP, which encodes the Synphilin-1 protein, was expressed in Saccharomyces cerevisiae and Schizosaccharomyces pombe yeast strains devoid of enzymes Glo1, Glo2, and Gre3. Presented data shows that lack of Glo2 and Gre3 activity in S. cerevisiae increases the formation of large Synphilin-1 inclusions. This correlates with enhanced oxidative stress levels and an inhibitory effect on exponential growth, which is most likely caused by deregulation of autophagic degradation capacity, due to excessive Synphilin-1 aggresome build-up. These findings illustrate the detrimental impact of increased oxidation and glycation on Synphilin-1 inclusion formation. Similarly, polar-localised inclusions were observed in wild-type S. pombe cells and strains deleted for either glo1+ or glo2+. Contrary to S. cerevisiae, however, no growth defects were observed upon expression of SNCAIP. Altogether, our findings show the relevance of yeasts, especially S. cerevisiae, as complementary models to unravel mechanisms contributing to Synphilin-1 pathology in the context of neurodegenerative diseases.

Supramolecular self-associating amphiphiles (SSAs) as nanoscale enhancers of cisplatin anticancer activity.

Many chemotherapeutic drugs have a narrow therapeutic window due to inefficient tumour cell permeation. Supramolecular self-associating amphiphilic salts (SSAs) are a unique class of small molecules that offer potential as next generation cancer drugs and/or therapeutic enhancement agents. Herein, we demonstrate the cytotoxicity of seven SSAs towards both ovarian and glioblastoma cancer cells. We also utilize the intrinsic fluorescent properties of one of these lead SSAs to provide evidence for this class of compound to both bind to the exterior cancer cell surface and permeate the cell membrane, to become internalized. Furthermore, we demonstrate synergistic effects of two lead SSAs on cisplatin-mediated cytotoxicity of ovarian cancer cells and show that this correlates with increased DNA damage and apoptosis versus either agent alone. This work provides the first evidence that SSAs interact with and permeate cancer cell membranes and enhance the cytotoxic activity of a chemotherapeutic drug in human cancer cells.

Towards the Prediction of Antimicrobial Efficacy for Hydrogen Bonded, Self-Associating Amphiphiles.

Herein we report 50 structurally related supramolecular self-associating amphiphilic (SSA) salts and related compounds. These SSAs are shown to act as antimicrobial agents, active against model Gram-positive (methicillin-resistant Staphylococcus aureus) and/or Gram-negative (Escherichia coli) bacteria of clinical interest. Through a combination of solution-state, gas-phase, solid-state and in silico measurements, we determine 14 different physicochemical parameters for each of these 50 structurally related compounds. These parameter sets are then used to identify molecular structure-physicochemical property-antimicrobial activity relationships for our model Gram-negative and Gram-positive bacteria, while simultaneously providing insight towards the elucidation of SSA mode of antimicrobial action.