Probe Design Research Articles

Exploring the Effects of Augmented Reality on the Emotion Recognition Skills of Autistic Children

Autistic children have significant difficulties in recognizing and labeling emotions, difficulties which may hinder the natural development of their ability to express and regulate further emotions. The purpose of this study was to evaluate the effect of an augmented reality emotion recognition program (ARERP) delivered in a group format on the emotion recognition of young autistic children. This study employed a concurrent multiple probe design across behaviors to evaluate the effects of the training on children’s emotion recognition. The target behavior was recognizing three emotion sets: positive, negative, and complex. The sequence of conditions included pre-test, baseline, training, post-test, and follow-up sessions. Positive results were found across all four children who all achieved 100% accuracy in identifying emotions during the training. Further, the emotion recognition skills were successfully maintained for 4–7 weeks post-training, with observed generalization to untrained still pictures and GIFs. This study provides preliminary evidence supporting the effectiveness of AR-based group training for young autistic children.

Enantioselective Synthesis of Chiral Sulfonimidoyl Fluorides Facilitates Stereospecific SuFEx Click Chemistry.

Sulfur-centered electrophilic 'warheads' have emerged as key components for chemical proteomic probes through sulfur-exchange chemistry (SuFEx) with protein nucleophiles. Among these functional groups, sulfonimidoyl fluorides (SIFs) stand out for their modifiable sites, tunable electrophilicities, and chiral sulfur-center, presenting exciting possibilities for new covalent chemical probes. However, the synthetic access to chiral SIFs has been a challenge, limiting their exploration and applications. In this study, we describe a convenient route to obtain chiral SIFs from readily available sulfenamides via a series of one-pot tandem reactions with high enantiomeric excess (ees). The resulting chiral SIFs were further converted into a diverse array of chiral S(VI) derivatives under mild conditions or in buffer solutions. Most significantly, the specificity of the chiral SIFs in protein ligation experiments underscored the critical role of sulfur-center chirality in the design and screening of more-selective covalent probes and therapeutics.

Rapid-response RNA-fluorescence in situ hybridization (FISH) assay platform for coronavirus antiviral high-throughput screening

Over the past 25 years, the global community has faced challenges posed by three distinct outbreaks of coronaviruses including severe acute respiratory syndrome coronavirus (SARS-CoV), Middle East respiratory syndrome coronavirus (MERS-CoV), and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The identification of a novel alphacoronavirus canine CoV (CCoV-HuPn2018) in human patients in Malaysia underscores the potential for crossover infections to humans. The threat of the ever-evolving nature of viral infections as well as the lingering health and socioeconomic effects of the recent SARS-CoV-2 pandemic emphasize the urgent need for advanced antiviral drug screening tools that can be quickly implemented to strengthen preparedness and preventive measures against future outbreaks. Here, we present the development and validation of a novel RNA-fluorescence in situ hybridization (FISH) imaging assay as a 384-well, high-throughput rapid response platform for antiviral drug discovery. RNA-FISH is a powerful tool to visualize specific mRNA in cultured cells using a high-content imaging platform. The flexibility of RNA-FISH probe sets allows for the rapid design of viral genome-specific probes, enabling in vitro assay development to test for inhibition of viral replication by either biologic or small molecule inhibitors. Screening of 170 antiviral compounds in concentration-response demonstrates a strong correlation between the RNA-FISH assay and an immunofluorescence assay (IFA) for both human coronaviruses HCoV-OC43 and HCoV-229E. Additionally, we successfully applied this methodology in the context of CCoV strain 1–71, proving rapid development and deployment, opening new avenues for the evaluation of antiviral drugs to potential future emerging threats.

Insights into the Structure and Dynamics of Proteins from 19F Solution NMR Spectroscopy.

19F NMR spectroscopy has recently witnessed a resurgence as an attractive analytical tool for the study of the structure and dynamics of biomolecules in vitro and in cells, despite reports of its applications in biomolecular NMR since the 1970s. The high gyromagnetic ratio, large chemical shift dispersion, and complete absence of the spin 1/2 19F nucleus from biomolecules results in background-free, high-resolution 19F NMR spectra. The introduction of 19F probes in a few selected locations in biomolecules reduces spectral crowding despite its increased line width in comparison to typical 1H NMR line widths and allows rapid site-specific measurements from simple 1D spectra alone. The design and synthesis of novel 19F probes with reduced line widths and increased chemical shift sensitivity to the surrounding environment, together with advances in labeling techniques, NMR methodology, and hardware, have overcome several drawbacks of 19F NMR spectroscopy. The increased interest and widespread use of 19F NMR spectroscopy of biomolecules is gradually establishing it as a sensitive and high-resolution probe of biomolecular structure and dynamics, supplementing traditional 13C/15N-based methods. This Review focuses on the advances in 19F solution NMR spectroscopy of proteins in the past 5 years, with an emphasis on novel 19F tags and labeling techniques, NMR experiments to probe protein structure and conformational dynamics in vitro, and in-cell NMR applications.

Design and Application of Nuclear and Molecular Theragnostic Probes

AbstractWith growing propensity towards personalised medication, the application and impact of molecular theragnostic (or, theranostic) is expected to increase dramatically. Molecular theragnostic is the combination of diagnostics and therapeutics in single targeted molecule. If successful, molecular theragnostic will determine the treatment strategy and therapeutic outcome apart from favourably influencing the time of treatment and comfort of the patient. Recent developments such as [64Cu]Cu‐ATSM for imaging tumor‐hypoxia, 99Y silicate for arthritis, 1⁷⁷Lu‐PSMA‐617 for prostate cancer etc has been discussed in detail in the literature.[1,2] This sectioned review deals in particular with the development of nuclear molecular theragnostic agents. The genesis and design strategies are presented after compiling recent reports.

Design and synthesis of naphthalimide-based CO fluorescent probes realizing tricolor detection and turn-on/off response

Carbon monoxide (CO) fluorescent probes have attracted tremendous attention attributed to their low detection limit, high specificity, simple operation and good biocompatibility. However, the relationship between the molecular structure and the fluorescence response signal still needs to be systematically elucidated for different applications. Herein, three different CO fluorescent probes based on naphthalimide derivative with blue emission (B-NIA), green emission (G-NIA), and near-infrared emission (R-NIA) were designed and synthesized. Three-primary colors and opposite turn-on or turn-off fluorescent responses to CO were achieved by regulating the conjugation system of fluorophores and the position of fluorescent recognition group. The change of fluorescence response and sensing mechanism for CO detection was studied by theory calculation and mass spectrometry analysis. All the B/G/R-NIA possessed high sensitivity for CO detection. Notably, the R-NIA emerged with a noticeable NIR fluorescence response to CO with a low detection limit (0.61 µM) and high selectivity and relative pH stability. In addition, the R-NIA showed particularly low cytotoxicity and has been successfully used to detect CO in living cells. These studies provided the theoretical reference and technical route to synthesize fluorescent probes with different emission wavelengths and opposite fluorescent responses to CO for various application scenarios

Development and optimization of the qPCR-based rapid detection method for Chinese species of Sanghuangporus

Sanghuangporus, well known as ‘Sanghuang’, is a macrofungal genus accommodating 11 edible and medicinal species in China. With the prosperous development of ‘Sanghuang’ business in China, the fraudulent practice naming the products of other morphologically and phylogenetically related species as ‘Sanghuang’ happens frequently, which disturbs the market order of ‘Sanghuang’ business. To promote the healthy industrial development of Sanghuangporus and also prevent the loss of this macrofungal resource, we develop a qPCR-based rapid detection method in this study. Internal transcribed spacer sequences of Fuscoporia, Inonotus, Phellinus, Sanghuangporus and Tropicoporus were aligned to design Sanghuangporus-specific amplification primers and probe, which can specifically distinguish all 11 Chinese species of Sanghuangporus from species of other closely related genera. The concentrations of Sanghuangporus-specific primers and probe were optimized as 0.7 μmol/L and 0.5 μmol/L, respectively, where the qPCR has the minimum detectable DNA template concentration of 10-3 ng/μL. In summary, the current qPCR-based detection method was specific, sensitive and rapid for Chinese species of Sanghuangporus.

Teaching students with autism spectrum disorder how to use an augmented reality application and the vocabulary of the natural sciences

Technology-based interventions are considered evidence-based for teaching students with Autism Spectrum Disorder (ASD). The main purpose of this study is to teach students with ASD the use of augmented reality (AR) mobile applications through systematic instruction and to test the effectiveness of the presented AR mobile application in teaching science concepts. The secondary aim of this study was to examine the students’ and teachers’ views on the acceptability of the AR mobile application in teaching science vocabulary. Three primary and one secondary school students with ASD, aged 10-13 years, participated in the study. A multiple probe design across participants was used to evaluate the effects of the intervention package. The findings showed that systematic instruction using an AR mobile application is effective in science vocabulary for students with ASD. All students reported using an AR mobile application to learn science vocabulary words was socially acceptable. These results suggest that the AR mobile applications may be a promising instruction instrument that is effective, efficient, and cost-effective. Implications for future research and practice are then discussed.

Effectiveness of a Three-Step Decoding Strategy with Constant Time-Delay to Teach Word Reading to Students with Mild and Moderate Intellectual Disability

The purpose of this study was to examine the effects of a three-step decoding strategy with constant time delay on teaching and acquisition of decoding and word reading skills to four students with mild and moderate intellectual disability and to measure the generalization of those skills to untaught words. A multiple probe design across participants was used to examine the percentage of sounds correctly decoded and words correctly read. The results indicate that all four participants learned to decode and read words using the two strategies. Furthermore, all participants showed an increased learning efficiency in decoding and reading when teaching the second set of similar words. Generalization data showed that only two participants were able to decode and read some of the new words before teaching them, while all participants' ability to decode the letters of the new words was improved. Keywords: Reading, Phonics, Three-step decoding, Time delay, Intellectual disability

Quantitative estimation of optical properties in bilayer media within the subdiffusive regime using tilted fiber-optic probe diffuse reflectance spectroscopy, part 2: probe design, realization, and experimental validation.

Tissues like skin have a layered structure where each layer's optical properties vary significantly. However, traditional diffuse reflectance spectroscopy assumes a homogeneous medium, often leading to estimations that reflects the properties of neither layer. There's a clear need for probes that can precisely measure the optical properties of layered tissues. This paper aims to design a diffuse reflectance probe capable of accurately estimating the optical properties of bilayer tissues in the subdiffusive regime. Using Monte Carlo simulations, we evaluated key geometric factors-fiber placement, tilt angle, diameter, and numerical aperture-on optical property estimation, following the methodology in Part I. A robust design is proposed that balances accurate intrinsic optical property (IOP) calculations with practical experimental constraints. The designed probe, featuring eight illumination and eight detection fibers with varying spacings and tilt angles. The estimation error of the IOP calculation for bilayer phantoms is less than 20% for top layers with thicknesses between 0.2 and 1.0mm. Building on the approach from Part I and using a precise calibration, the probe effectively quantified and distinguished the IOPs of bilayer samples, particularly those relevant to early skin pathology detection and characterization.

Rationally Designed G-Quadruplex Selective "Turn-On" NIR Fluorescent Probe with Large Stokes Shift for Nucleic Acid Research-Based Applications.

Guanine-rich DNA/RNA sequences can form Hoogsteen bonds to adopt noncanonical secondary structures called G-quadruplexes, and these have been associated with diverse cellular processes. There has been considerable research interest in the design of G4-interacting ligands for cellular probing of the G4 structure and understanding its associated biological function. Most of the fluorescent G4 ligands either do not have significant selectivity over other nucleic acid structures, have high Stokes shift, or are not in the near-infrared (NIR) region, which limits its cellular visualization. The current work involves the rational design and synthesis of NIR fluorescent probes comprising a (Z)-1-methyl-2-((3-methylbenzo[d]thiazol-2(3H)-ylidene)methyl)quinolin-1-ium scaffold. Among the designed molecules, 4a exhibited far-red fluorescence (λmax = 680 nm) with large Stokes shift (∼182 nm) upon selective binding to human telomeric G-quadruplexes. The dye 4a does not disturb the conformation and stability of G-quadruplexes, thereby making it suitable for nucleic acid research based applications. Interestingly, 4a showed remarkable selectivity over single- and double-stranded structures in contrast to a commercially available quadruplex binding probe, Thiazole orange (TO). The molecular docking studies indicate that 4a binds at the groove region of the telomeric DNA G-quadruplex through π-π stacking interactions with the quinoline and amine-substituted phenyl ring and with the phosphate backbone through anion-π interactions with the benzothiazole ring. The designed molecule 4a has interesting photophysical properties, cell permeability, and biocompatibility with minimal cytotoxicity. Fluorescence imaging studies in live HeLa cells showed that probe 4a binds to the transient population of the DNA G-quadruplex in the nucleus and RNA quadruplexes in the cytoplasm. In brief, G-quadruplex NIR fluorescent probe 4a with a higher signal/noise ratio has significant potential for cellular imaging studies and thus opens avenues to decipher the biological pathways for better understanding of G-quadruplex biology.

Efficient and reliable corrosion control for subsea assets: challenges in the design and testing of corrosion probes in aggressive marine environments

Abstract This review discusses the challenges in designing and testing corrosion probes for aggressive marine environments. The objectives are to analyze existing literature, identify methodological problems, and highlight research gaps in subsea corrosion control. To achieve these, a comprehensive review of relevant literature was conducted, focusing on factors like high salinity, fluctuating temperatures, and the presence of corrosive agents. The methods involved synthesizing information from peer-reviewed articles, industry reports, and academic publications to thoroughly analyze current state of knowledge. The findings of this review highlight the need for standardized testing protocols, improved understanding of material compatibility, and consideration of real-world conditions in corrosion probe design and testing. Methodological problems include the lack of standardized testing protocols, limited understanding of material compatibility, and insufficient consideration of real-world conditions. These findings emphasize the challenges researchers and practitioners face in developing efficient and reliable corrosion control strategies for subsea assets. In terms of novelty and improvement, this manuscript contributes to improving corrosion control practices in aggressive marine environments by synthesizing existing literature, identifying methodological problems, and highlighting gaps. By addressing these challenges, future research can focus on developing innovative solutions and methodologies to enhance the durability and effectiveness of corrosion probes in subsea environments.

Near-Infrared Unimolecular Chemiluminescence Probes for Deep-Tissue Imaging.

Chemiluminescence (CL) imaging has emerged as a promising optical imaging technique due to minimal background autofluorescence and being excitation-free. However, the emission of most chemiluminescent probes was concentrated in the visible light region, which limited the tissue penetration. Although some NIR chemiluminescence probes have been reported based on the chemiluminescence resonance energy transfer (CRET) strategy, the energy loss was inevitable. Thus, it is crucial to develop near-infrared (NIR) unimolecular probes with direct chemiluminescence. Herein, we propose a strategy of increasing conjugation for designing and synthesizing novel NIR chemiluminescence unimolecular probes that consist of luminol, electron acceptor, π-bridge, and electron donor. Luminol was conjugated to the unimolecular backbone to produce direct NIR chemiluminescence. Notably, the direct CL mechanism of probes was investigated. Compared with CRET-based chemiluminescence, this direct CL was more advantageous to immediately convert the chemical energy into chemiluminescence, avoiding energy degradation. Furthermore, the corresponding nanoparticles with great biosafety were prepared by self-assembly with amphiphilic DSPE-PEG. Especially, TTBL@PEG-NPs with NIR-I emission were successfully used in the sensitive in vivo chemiluminescence imaging of various inflammation models, such as peritonitis, ear swelling, and colitis. This study paves the way for the design of NIR unimolecular chemiluminescence probes and deep-tissue imaging.

Lanthanide-Doped Nanoparticles Anchoring on Metal-Organic Frameworks with Thermally Enhanced Upconversion Luminescence for Sensitive Nanothermometers.

Nanothermometers can detect changes in the local temperature in living cells and in vivo, revealing fundamental biological properties. Despite the exploration of different temperature-responsive materials, the design and development of temperature-sensing probes with high brightness and high sensitivity remain a daunting challenge. Here, we employed the UiO-66 type metal-organic frameworks (MOFs) to anchor UNCPs on the surface of the MOFs for constructing MOF@UCNPs nanohybrids. The in situ composite method with MOFs leads to the coordination interaction between the ligands and the surface of UCNPs, enabling controlled composite formation between different MOFs and UCNPs. Remarkably, the surface interaction favors the anomalous thermo-enhanced luminescence, achieving a 35-fold enhancement of UiO-66@NaYF4:Yb/Tm at 413 K. Furthermore, these MOF@UCNPs nanohybrids with thermo-enhanced luminescence are developed as multifunctional biological probes for bioimaging and intracellular temperature sensing, demonstrating a high thermal sensitivity of 1.92% K-1 in the physiological temperature range. Based on these findings, temperature monitoring of the local position was successfully carried out by the designed MOF@UCNPs nanoprobes in vivo. These findings underscore the potential of MOF@UCNPs nanohybrids, opening up new avenues for the development of a multifunctional platform for biological analysis.

Investigation on Understanding the Numeracy Capacity of Intellectual Disabled Students using Enabling Technology Tools: Web Application, AR and UI/UX

The population of individuals with intellectual disabilities (ID) is increasing, necessitating assistance with a wide range of daily activities. Acquiring and assessing numeracy and communication skills are critical for this demographic, requiring tools and techniques tailored to their specific needs. Effective educational tools must employ multi-modal and multi-sensory approaches to cater to diverse learning styles and incorporate assistive technological solutions. Despite the availability of numerous tools, there is a need to enhance their utility and effectiveness. This study aims to identify and refine the requirements for an innovative educational tool that employs Two-dimensional (2D) and Augmented Reality (AR) technologies. To achieve this, we conducted semi-structured interviews and surveys with teachers working with students with ID, gaining insights into the current solutions, advantages, and limitations. Additionally, we used physical props as design probes in a co-design methodology to better understand and elicit the true needs of individuals with ID. The findings from this research will inform the development of a 2D/AR tool designed to make learning mathematics more engaging and effective for individuals with ID, contributing to the advancement of inclusive education practices. Enabling Technology plays a significant role in the numeracy ability among people with ID. Generative AI and Explainable AI shall further improve learning ability in the years to come.

Polydimethylsiloxanes - Based Fluorescent Probe for H2S Detection in Living Cells.

The development of fluorescent probes for H2S detection especially in living cells is of great significance due to its fundamental role as signal molecule. A promising scaffold for the development of such probes is polydimethylsiloxanes (PDMS), which is cost-effectiveness, non-toxicity, flexibility, and biocompatibility and easy to post-functionalize. Surprisingly, fluorescent probes for H2S detection based on PDMS have not been investigated. Moreover, 4-nitro - 2,1,3-benzoxadiazole (NBD) derivates provides high fluorescence quantum yield, a large molar absorption coefficient, and sensitivity to environmental changes. Through reasonable design and adjustment of substituents on the NBD group, precise control of its fluorescence properties can be achieved. Herein, a novel H2S fluorescent probe, P-NBD, was designed and synthesized by a one-step aromatic ring nucleophilic substitution of Cl-NBD with a PDMS derivative. Due to the occurrence of the cleavage reaction strategy and the intramolecular charge transfer process, P-NBD can detect H2S via a colorimetric method. The limit of detection is down to 93 nM. P-NBD demonstrated considerable detection capability comparable to other reported types of H2S probes. Moreover, the probe can also be utilized for H2S imaging in HeLa cells. This work provides new insights into the design and synthesis of novel H2S probes while also offering experimental evidence for the application of PDMS in cellular imaging.

A "Goldilocks Zone" for Recruiting BET Proteins with Bromodomain-1-Selective Ligands.

Synthetic genome readers/regulators (SynGRs) are bifunctional molecules that are rationally designed to bind specific genomic sequences and engage cellular machinery that regulates the expression of targeted genes. The prototypical SynGR1 targets GAA trinucleotide repeats and recruits the BET family of transcriptional regulatory proteins via a flexibly tethered ligand, JQ1. This pan-BET ligand binds both tandem bromodomains of BET proteins (BD1 and BD2). Second-generation SynGRs, which substituted JQ1 with bromodomain-selective ligands, unexpectedly revealed that BD1-selective ligands failed to functionally engage BET proteins in living cells despite displaying the ability to bind BD1 in vitro. Mechanistically, recruiting a BET protein via BD1- or BD2-selective SynGRs should have resulted in indistinguishable functional outcomes. Here we report the conversion of inactive BD1-targeting SynGRs into functional gene regulators by a structure-guided redesign of the chemical linker that bridges the DNA-binding molecule to the highly selective BD1 ligand GSK778. The results point to an optimal zone for positioning the BD1-selective ligand for functional engagement of BET proteins on chromatin, consistent with the preferred binding of BD1 domains to distal acetyllysine residues on histone tails. The results not only resolve the mechanistic conundrum but also provide insight into domain-selective targeting and nuanced design of chemo probes and therapeutics.

A Macrocyclic Hybrid PET/MRI Probe for Quantitative Perfusion Imaging In Vivo.

Perfusion dynamics play a vital role in delivering essential nutrients and oxygen to tissues while removing metabolic waste products. Imaging techniques such as magnetic resonance imaging (MRI), computed tomography (CT), and positron emission tomography (PET) use contrast agents to visualize perfusion and clearance patterns; however, each technique has specific limitations. Hybrid PET/MRI combines the quantitative power and sensitivity of PET with the high functional and anatomical detail of MRI and holds great promise for precision in molecular imaging. However, the development of dual PET/MRI probes has been hampered by challenging synthesis and radiolabeling. Here, we present a novel PET/MRI probe, [18F][Gd(FL1)], which exhibits excellent stability comparable to macrocyclic MRI contrast agents used in clinical practice. The unique molecular design of [18F][Gd(FL1)] allows selective and expeditious radiolabeling of the gadolinium chelate in the final synthetic step. Leveraging the strengths of MRI and PET signals, the probe enables quantitative in vivo mapping of perfusion and excretion dynamics through an innovative voxel-based analysis. The diagnostic capabilities of [18F][Gd(FL1)] were demonstrated in a pilot study on healthy mice, successfully detecting early cases of unilateral renal dysfunction, a condition that is typically challenging to diagnose. This study introduces a new approach for PET/MRI and emphasizes a streamlined probe design for practical synthesis and improved diagnostic accuracy.

Circumferential Crack Detection in Ultra-High-Pressure Tubular Reactors with Pulsed Eddy Current Testing.

Ultra-high-pressure tubular reactors are crucial pieces of equipment for polyethylene production. Long-term operation under high temperature, high pressure, and other extremely harsh conditions can lead to various defects, with circumferential cracks posing a major safety risk. Detecting cracks is challenging, particularly when they are under a protective layer of a certain thickness. This study designed a pulsed eddy current differential probe to detect circumferential cracks in ultra-high-pressure tubular reactors, with the lift-off distance acting as a protective layer. Detection models for traditional cylindrical and semi-circular excitation differential probes were established using finite element simulations. Corresponding experiments under different lift-off conditions were carried out, and the model's accuracy was verified by the consistency between the simulation results and experimental data. The distribution of the eddy current field under different conditions and the disturbances caused by cracks at various positions to the detection signal were then calculated in the simulations. The simulation results showed that the cracks significantly disturbed the eddy current field of the semi-circular excitation differential probe compared with that of the traditional cylindrical probe. The designed differential probe effectively detected circumferential cracks of specific lengths and depths using the difference in the voltage signals. The experimental results were in agreement with the simulation results, showing that the designed probe could effectively detect 20 mm-long circumferential cracks at a lift-off of 60 mm. The experimental results also show that the probe's detection coverage area in the axial direction varied with the lift-off height. The probe design and findings are valuable for detecting cracks in ultra-high-pressure tubular reactors with protective layers.

Design and Synthesis of Small Molecule Probes of MDA-9/Syntenin.

MDA-9/Syntenin, a key scaffolding protein and a molecular hub involved in a diverse range of cell signaling responses, has proved to be a challenging target for the design and discovery of small molecule probes. In this paper, we report on the design and synthesis of small molecule ligands of this key protein. Genetic algorithm-based computational design and the five-eight step synthesis of three molecules led to ligands with affinities in the range of 1-3 µM, a 20-60-fold improvement over literature reports. The design and synthesis strategies, coupled with the structure-dependent gain or loss in affinity, afford the deduction of principles that should guide the design of advanced probes of MDA-9/Syntenin.