Single Probe Research Articles

A Single Electrochemical Biosensor Designed to Detect Any Virus.

Viruses are the primary cause of many infectious diseases in both humans and animals. Various testing methods require an amplification step of the viral RNA sample before detection, with quantitative reverse transcription polymerase chain reaction (RT-qPCR) being one of the most widely used along with lesser-known methods like Nucleic Acid Sequence-Based Amplification (NASBA). NASBA offers several advantages, such as isothermal amplification and high selectivity for specific sequences, making it an attractive option for low-income facilities. In this research, we employed a single electrochemical biosensor (E-Biosensor) designed for potentially detecting any virus by modifying the NASBA protocol. In this modified protocol, a reverse primer is designed with an additional 22-nucleotide sequence (tag region) at the 5'-end, which is added to the NASBA process. This tag region becomes part of the final amplicon generated by NASBA. It can hybridize with a single specific E-Biosensor probe set, enabling subsequent virus detection. Using this approach, we successfully detected three different viruses with a single E-Biosensor design, demonstrating the platform's potential for virus detection.

Cosmography with supernova Refsdal through time-delay cluster lensing: Independent measurements of the Hubble constant and geometry of the Universe

We present new measurements of the values of the Hubble constant, matter density, dark energy density, and dark energy density equation-of-state (EoS) parameters. These results have been obtained from a full strong-lensing analysis of the observed positions of 89 multiple images and 4 measured time delays of the supernova (SN) Refsdal in the Hubble Frontier Fields galaxy cluster MACS J1149.5+2223. By strictly following the identical modelling methodology (as done in our previous work undertaken before time delays were available), our cosmographic measurements are essentially blind, based on the frozen procedure. Without using any priors from other cosmological experiments, in an open wCDM cosmological model and via our reference cluster mass model, we measure the following values: H0 = 65.1−3.4+3.5 km s−1 Mpc−1, ΩDE = 0.76−0.10+0.15, and w = −0.92−0.21+0.15 (at the 68.3% confidence level). No other single cosmological probe has been able to simultaneously measure all these parameters. Remarkably, our estimated values of the cosmological parameters, in particular that of H0, are very robust and do not significantly depend on the assumed cosmological model or the cluster mass modelling details. The latter aspect introduces systematic uncertainties on the values of H0 and w, which are found to be largely subdominant compared to the statistical errors. The results of this study demonstrate that the combination of time delays in lens galaxy clusters with extensive photometric and spectroscopic information offers a novel and competitive cosmological tool.

Multimodal fiber probe for simultaneous mid-infrared and Raman spectroscopy

A fiber probe has been developed that enables simultaneous acquisition of mid-infrared (MIR) and Raman spectra in the region of 3100–2600 cm−1. Multimodal measurement is based on a proposed ZrO2 crystal design at the tip of an attenuated total reflection (ATR) probe. Mid-infrared ATR spectra are obtained through a pair of chalcogenide infrared (CIR) fibers mounted at the base of the crystal. The probe enables both excitation and acquisition of a weak Raman signal from a portion of the sample in front of the crystal using an additional pair of silica fibers located in a plane perpendicular to the CIR fibers. The advantages of combining MIR and Raman spectra in a single probe have been discussed.

Lanthanide‐Based Probes for Imaging Detection of Enzyme Activities by NIR Luminescence, T1‐ and ParaCEST MRI

AbstractApplying a single molecular probe to monitor enzymatic activities in multiple, complementary imaging modalities is highly desirable to ascertain detection and to avoid the complexity associated with the use of agents of different chemical entities. We demonstrate here the versatility of lanthanide (Ln3+) complexes with respect to their optical and magnetic properties and their potential for enzymatic detection in NIR luminescence, CEST and T1 MR imaging, controlled by the nature of the Ln3+ ion, while using a unique chelator. Based on X‐ray structural, photophysical, and solution NMR investigations of a family of Ln3+ DO3A‐pyridine model complexes, we could rationalize the luminescence (Eu3+, Yb3+), CEST (Yb3+) and relaxation (Gd3+) properties and their variations between carbamate and amine derivatives. This allowed the design of probes which undergo enzyme‐mediated changes detectable in NIR luminescence, CEST and T1‐weighted MRI, respectively governed by variations in their absorption energy, in their exchanging proton pool and in their size, thus relaxation efficacy. We demonstrate that these properties can be exploited for the visualization of β‐galactosidase activity in phantom samples by different imaging modalities: NIR optical imaging, CEST and T1‐weighted MRI.

Identification of ADME genes polymorphic variants linked to trastuzumab-induced cardiotoxicity in breast cancer patients: Case series of mono-institutional experience

BackgroundLong-term survival induced by anticancer treatments discloses emerging frailty among breast cancer (BC) survivors. Trastuzumab-induced cardiotoxicity (TIC) is reported in at least 5% of HER2+BC patients. However, TIC mechanism remains unclear and predictive genetic biomarkers are still lacking. Interaction between systemic inflammation, cytokine release and ADME genes in cancer patients might contribute to explain mechanisms underlying individual susceptibility to TIC and drug response variability. We present a single institution case series to investigate the potential role of genetic variants in ADME genes in HER2+BC patients TIC experienced. MethodsWe selected data related to 40 HER2+ BC patients undergone to DMET genotyping of ADME constitutive variant profiling, with the aim to prospectively explore their potential role in developing TIC. Only 3 patients (“case series”), who experienced TIC, were compared to 37 “control group” matched patients cardiotoxicity-sparing. All patients underwent to left ventricular ejection fraction (LVEF) evaluation at diagnosis and during anti-HER2 therapy. Each single probe was clustered to detect SNPs related to cardiotoxicity. ResultsIn this retrospective analysis, our 3 cases were homogeneous in terms of clinical-pathological characteristics, trastuzumab-based treatment and LVEF decline. We identified 9 polymorphic variants in 8 ADME genes (UGT1A1, UGT1A6, UGT1A7, UGT2B15, SLC22A1, CYP3A5, ABCC4, CYP2D6) potentially associated with TIC. ConclusionReal-world TIC incidence is higher compared to randomized clinical trials and biomarkers with potential predictive value aren’t available. Our preliminary data, as proof of concept, could suggest a predictive role of pharmacogenomic approach in the identification of cardiotoxicity risk biomarkers for anti-HER2 treatment.

Design of dual circular polarized stacked patch antenna for BeiDou navigation satellite systems

In this article, a design of two-layered, dual circular-polarization (CP) and dual-band path antenna is presented for BeiDou navigation satellite (BDS) system applications. A single 50-Ω Co-axial probe feeding arrangement is employed to achieve dual-band operation and it feeds both the antennas simultaneously. This mode introduces the CP radiation on both upper (RHCP) and lower (LHCP) operation bands. The impedance bandwidth is further widened by accommodating two semi-circular stubs on the top patch. The fabricated and experimentally validated prototype radiates at 1.82 and 3.45 GHz (center frequency). The upper and lower operating band exhibits right and left-handed CP radiations, which constructs dual-band operation. The fabricated prototype has a physical size of πr2 × 0.024 λg with an antenna profile of 2.38 mm (0.024 λg). The experimentally validated upper and lower bands impedance bandwidth with |S11| < −10 dB are 21.1% and 11.1%, respectively. The fabricated prototype is a good candidate to use in BDS applications. [Q2

A bias-free test of human temporal bisection: Evidence against bisection at the arithmetic mean

The temporal bisection procedure has been used to assess theories of time perception. A problem with the procedure for measuring the perceived midpoint of two durations is that the spacing of probe durations affects the length of the bisection point. Linear spacing results in longer bisection points closer to the arithmetic mean of the durations than logarithmic spacing. In three experiments, the influence of probe duration distribution was avoided by presenting a single probe duration of either the arithmetic or geometric mean of the trained durations. It was found that the number of participants that categorised the arithmetic mean as long was significantly larger than those that categorised it as short. The number of participants that categorised the geometric mean as either short or long did not significantly differ. This was true for trained durations of 0.4 s vs. 1.6 s (Experiments 1–3), 0.2 s vs. 3.2 s (Experiment 2) and 0.4 s vs. 6.4 s (Experiment 3). In Experiment 4, the probe trial distribution effect was replicated with logarithmic and linearly distributed probe durations, demonstrating that bisection occurs close to the arithmetic mean with linearly spaced probe durations. The results provide evidence against bisection at the arithmetic mean when probe spacing bias is avoided and, instead, the results are consistent with logarithmic encoding of time, or a comparison rule based on relative rather than absolute differences.

Abstract 7114: Exploring the landscape of PARP and PARG inhibitor selectivity in live cells using NanoBRET™ target engagement assays

Abstract The 17-member poly(ADP-ribose) polymerase (PARP) family of proteins mediate a number of cellular processes including gene transcription, cell-division and DNA-repair. PARPs catalyze the addition of poly- or mono- ADP-ribose chains on target proteins, modifying their function and/or recruitment in various pathways. Poly(ADP-ribose) Glycohydrolase (PARG) catabolizes PAR units and facilitates the recycling of PARP and other modified effector proteins. A number of PARPS are implicated in human disease, and PARP inhibitors (PARPi’s) have seen major clinical success, however, many PARPs (especially mono PARPs) are understudied and chemical tools for understanding cellular PARPi selectivity and potency across the entire family are lacking. Similarly, several PARG inhibitors have been developed to date, but tools for directly assessing target engagement in a cellular context are sorely needed. Here we describe the development and application of target engagement assays for characterizing PARP and PARG inhibitor selectivity and affinity in live cells. This method is based on the NanoBRET™ assay platform, which is a proximity-based probe displacement assay that operates in live cells. Using NanoBRET™ technology, we have developed a single probe that covers 12 out of 17 PARP family members. We explored the family-wide selectivity of a diverse panel of clinical and preclinical PARP inhibitors in live cells, representing the first family-wide, cell-based method for profiling PARP selectivity. We also describe the development of a second probe that reports on PARG target engagement. Target engagement of PARG was observed for three out of five tested inhibitors, suggesting a potential indirect mode of action for certain inhibitors on PARG glycohydrolase activity. These assays offer a complementary approach for studying target engagement within the PARylation cycle. Citation Format: Ani Michaud, Kelly Teske, Jennifer Wilkinson, Chad Zimprich, James Vasta, Cesear Corona, Min Zhou, Elizabeth Dominguez, Kaitlin Dunn-Hoffman, Matthew Robers. Exploring the landscape of PARP and PARG inhibitor selectivity in live cells using NanoBRET™ target engagement assays [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7114.

Lanthanide-Based Probes for Imaging Detection of Enzyme Activities by NIR Luminescence, T1- and ParaCEST MRI.

Applying a single molecular probe to monitor enzymatic activities in multiple, complementary imaging modalities is highly desirable to ascertain detection and to avoid the complexity associated with the use of agents of different chemical entities. We demonstrate here the versatility of lanthanide (Ln3+) complexes with respect to their optical and magnetic properties and their potential for enzymatic detection in NIR luminescence, CEST and T1 MR imaging, controlled by the nature of the Ln3+ ion, while using a unique chelator. Based on X-ray structural, photophysical, and solution NMR investigations of a family of Ln3+ DO3A-pyridine model complexes, we could rationalize the luminescence (Eu3+, Yb3+), CEST (Yb3+) and relaxation (Gd3+) properties and their variations between carbamate and amine derivatives. This allowed the design of probes which undergo enzyme-mediated changes detectable in NIR luminescence, CEST and T1-weighted MRI, respectively governed by variations in their absorption energy, in their exchanging proton pool and in their size, thus relaxation efficacy. We demonstrate that these properties can be exploited for the visualization of β-galactosidase activity in phantom samples by different imaging modalities: NIR optical imaging, CEST and T1-weighted MRI.

Impact of High-Pressure-Turbine Purge Flow on the Evolution of Turbulence in a Turbine Vane Frame

Abstract This paper describes the measurement and postprocessing of turbulence data. The experiments were carried out in a two-stage, two-spool transonic turbine test rig at the Institute for Thermal Turbomachinery and Machine Dynamics at the Graz University of Technology, which includes relevant purge and turbine rotor tip leakage flows. The test setup consists of a high-pressure turbine (HPT) stage, a turbine vane frame (TVF) with turning struts and splitters, and a counter-rotating low-pressure turbine (LPT) to allow engine realistic measurements. Time-resolved area traverse measurements have been performed for three different operating conditions in three measurement planes downstream of the HPT rotor, which enable the measurement of the turbulence quantities at the TVF inlet and outlet as well as the LPT outlet. The turbulence quantities are evaluated using triaxial- and single hot-film probes by means of Constant-Temperature-Anemometry, and their results were validated with five-hole probe (FHP) measurements. Ensemble- and time-averaging, as well as Fourier transforms, were applied for data reduction. It is shown how the turbulence intensity and integral length scale vary over different purge flowrates (PFR). The acquired measurement data illustrate that the interaction of the ejected purge flow with mean flow enhances the turbulent mixing in the secondary flow structures at the TVF inlet and TVF outlet, respectively. Furthermore, the flow downstream of the LPT rotor is affected by TVF and LPT rotor secondary flow structures, which are identified as high turbulence regions. These regions strongly depend on the purge flowrate. These results acquired under engine realistic rig flow conditions enable a deeper understanding of the relationship between loss and turbulence quantities and will help improve the application of computational fluid dynamics turbulence models to turbomachinery modeling.

GNAS AS2 methylation status enables mechanism-based categorization of pseudohypoparathyroidism type 1B.

Pseudohypoparathyroidism type 1B (PHP1B) results from aberrant genomic imprinting at the GNAS gene. Defining the underlying genetic cause in new patients is challenging because various genetic alterations (e.g., deletions, insertions) within the GNAS genomic region, including the neighboring STX16 gene, can cause PHP1B, and the genotype-epigenotype correlation has not been clearly established. Here, by analyzing patients with PHP1B with a wide variety of genotypes and epigenotypes, we identified a GNAS differentially methylated region (DMR) of distinct diagnostic value. This region, GNAS AS2, was hypomethylated in patients with genetic alterations located centromeric but not telomeric of this DMR. The AS2 methylation status was captured by a single probe of the methylation-sensitive multiplex ligation-dependent probe amplification (MS-MLPA) assay utilized to diagnose PHP1B. In human embryonic stem cells, where NESP55 transcription regulates GNAS methylation status on the maternal allele, AS2 methylation depended on 2 imprinting control regions (STX16-ICR and NESP-ICR) essential for NESP55 transcription. These results suggest that the AS2 methylation status in patients with PHP1B reflects the position at which the genetic alteration affects NESP55 transcription during an early embryonic period. Therefore, AS2 methylation levels can enable mechanistic PHP1B categorization based on genotype-epigenotype correlation and, thus, help identify the underlying molecular defect in patients.

Reversible Dual Fluorescence-Lifetime Imaging of Mitochondrial GSH and Microviscosity: Real-Time Evaluation of Ferroptosis Status.

Ferroptosis, as a new form of regulated cell death, is implicated in various physiological and pathological processes. Developing a single probe for an independent analysis of multiple analytes related to ferroptosis can provide more accurate information and simplify the detection procedures, but it faces great challenges. In this work, we develop a fluorescent probe for the simultaneous detection of GSH through ratiometric fluorescence response and microviscosity via a fluorescence lifetime model. Based on the reversible Michael addition reaction between GSH and unsaturated C═C bond, the probe responds reversibly to GSH with a ratiometric fluorescence variation and a fast response time (t1/2 = 4.7 s). At the same time, the probe is sensitive to environmental viscosity by changing its fluorescence lifetimes. The probe was applied to monitor the drug-induced ferroptosis process through both the classical Xc-/GSH/GPX4- and DHODH-mediated defense mechanisms. We hope that the probe will provide a useful molecular tool for the real-time live-cell imaging of GSH dynamics, which is benefit to unveiling related physiological and pathological processes.

Single Fluorescent Probe for Multiple Tasks: Illuminating Lipid Droplets and Lysosomes in Dual Channels and Distinguishing Autophagy and Apoptosis.

Lipid droplets (LDs) and lysosomes play key roles in autophagy and cell apoptosis, and the discriminative visualization of the two organelles and simultaneously of autophagy and apoptosis is very helpful to understand their internal relationships. However, fluorescent probes that can concurrently achieve these tasks are not available currently. Herein, we delicately fabricate a robust probe CAQ2 for multiple tasks: illumination of LDs and lysosomes in dual emission colors as well as discriminative visualization of cell apoptosis and autophagy. The probe exhibited both lipophilic and basic properties and displayed different emission colors in neutral and protonated forms; thus, LDs and lysosomes emitted blue and red fluorescence colors, respectively. Because of the lysosomal acidification during autophagy, CAQ2 detected autophagy with evidently enhanced red emission. Because of the lysosomal alkalization during apoptosis, CAQ2 imaged apoptosis with a drastically decreased red fluorescence intensity. With the robust probe, the autophagy under starvation and lipidless conditions was visualized, and the apoptosis induced by H2O2, ultraviolet (UV) irradiation, and rotenone treatment was successfully observed. The efficient detoxification of Na2S against rotenone treatment was successfully revealed.

Fisher information rates in sequentially measured quantum systems

We consider the impact that temporal correlations in the measurement statistics can have on the achievable precision in a sequential metrological protocol. In this setting, and for a single quantum probe, we establish that it is the transitions between the measurement basis states that plays the most significant role in determining the precision, with the resulting conditional Fisher information being interpretable as a rate of information acquisition. Projective measurements are shown to elegantly demonstrate this in two disparate estimation settings. Firstly, in determining the temperature of an environment and, secondly, to ascertain a parameter of the system Hamiltonian. In both settings we show that the sequential estimation approach can provide a useful method to enhance the achievable precision.

A field programmable gate array based Langmuir probe system for measurement of plasma parameters at 500kHz in a high-power impulse magnetron sputtering plasma.

By utilizing Field Programmable Gate Arrays in a configuration similar to that of the Mirror Langmuir Probe, it is possible to bias a single probe at three precise voltages in sequence. These voltages can be dynamically adjusted in real-time based on the measured plasma electron temperature to ensure the transition region is always sampled. The first results have been obtained by employing this method and have generated real-time outputs of electron temperature, ion saturation current, and floating potential on a low temperature pulsed-DC magnetron at 500kHz. These results are in good agreement with the analysis of a conventionally swept Langmuir probe. This probe is designed with the intention of being implemented on MAST-U to aid in the study of exhaust physics and enable further investigation into filamentary behavior.

Modeling and prediction of surface free energy of pavement materials based on a single probe liquid using artificial neural network

Surface free energy (SFE) is a vital material property employed for the assessment of various pavement performances such as asphalt-aggregate adhesion, moisture resistance, material compatibility, etc. However, the conventional models for estimating the SFE of materials require the contact angles (CA) of multiple probe liquids (PL) on the material's surface. In addition, the process requires further computational effort to arrive at the SFE components. In an attempt to offer an accurate and quicker alternative method of estimating the SFE of materials with lesser effort, this study has explored the use of artificial neural network (ANN) to predict the SFE of pavement materials. A simple but novel data structuring was employed to predict the SFE using inputs from just a single PL. Variables needed for the successful prediction of SFE components were identified and studied. Sensitivity analysis along with relative importance was used to rank the predicting variables. The Lifshitz-van der Waals (LW) component ANN-model showed the best accuracy (RMSE = 1.652 J/m2, R = 0.984). The base and acid components models showed relatively lower performances, (RMSE = 2.560 J/m2, R = 0.943) and (RMSE = 0.575 J/m2, R = 0.818) respectively. However, the SFE components estimated from the ANN-models yielded a total SFE with very good accuracy (RMSE = 2.049 J/m2, R = 0.982). Further insight into which variables such as substrate-type, CA test method, PL type, etc. facilitate model accuracy was provided.

Single-Response Duplexing of Electrochemical Label-Free Biosensor from the Same Tag.

Multiplexing is a valuable strategy to boost throughput and improve clinical accuracy. Exploiting the vertical, meshed design of reproducible and low-cost ultra-dense electrochemical chips, the unprecedented single-response multiplexing of typical label-free biosensors is reported. Using a cheap, handheld one-channel workstation and a single redox probe, that is, ferro/ferricyanide, the recognition events taking place on two spatially resolved locations of the same working electrode can be tracked along a single voltammetry scan by collecting the electrochemical signatures of the probe in relation to different quasi-reference electrodes, Au (0V) and Ag/AgCl ink (+0.2V). This spatial isolation prevents crosstalk between the redox tags and interferences over functionalization and binding steps, representing an advantage over the existing non-spatially resolved single-response multiplex strategies. As proof of concept, peptide-tethered immunosensors are demonstrated to provide the duplex detection of COVID-19 antibodies, thereby doubling the throughput while achieving 100% accuracy in serum samples. The approach is envisioned to enable broad applications in high-throughput and multi-analyte platforms, as it can be tailored to other biosensing devices and formats.

A difunctional fluorescent probe to detect hypochlorite and hydrogen sulfide ions in ferroptosis and oxidative stress induced by Aspirin

HOCl and H2S frequently coexist in living cells and work in concert to maintain the redox dynamic balance in vivo. It is challenging to precisely and simultaneously trace both HOCl and H2S using a single recognition-type fluorescence probe. Here, a novel fluorescent probe (MB-HS) for the selective detection of hydrogen sulphide (H2S) and hypochlorous acid (HOCl) has been established based on the coupling of the methylene blue fraction and the indolium fraction. For HOCl, the dual-signal probe MB-HS displays quick turn-on fluorescence responses. However, for H2S, it exhibits turn-off fluorescence at 586 nm and turn-on at 513 nm, respectively. MB-HS showed many advantages such as quick response, strong selectivity, and high sensitivity. It was effectively used to detect endogenous HOCl/H2S in ferroptosis and oxidative stress induced by Aspirin in HepG-2 cells.

Core ion measurements with collective Thomson scattering for DEMO burn control

DEMO burn control will require measurements of a range of plasma parameters, but the suite of feasible diagnostics for this purpose is limited. Here we assess the accuracy with which a collective Thomson scattering (CTS) diagnostic can provide key measurements for burn control in the planned European DEMO (EU-DEMO). This is based on estimated signal-to-noise ratios for a conceptual diagnostic design and trial fits to synthetic DEMO CTS spectra. We show that a diagnostic with a single probe- and receiver beam setup will be able to provide simultaneous measurements of core fusion alpha density and ion temperature to mean accuracies better than 5% and 10%, respectively, along with detecting intrinsic toroidal rotation velocities down to within ∼5 km s−1. Adding a second CTS receiver view furthermore enables inference of the core fuel-ion ratio, allowing discrimination between, e.g. a 50%/50% and 55%/45% D/T mixture, while also providing useful information on the thermalized He content. A DEMO CTS diagnostic would thus be able to monitor fusion alpha densities as well as anomalous transport of fast alphas and heat from the plasma core, quantify plasma rotation for confinement enhancement, and track the core isotope mix for optimum fusion performance. This versatility makes such a diagnostic a potentially valuable tool for real-time burn control on DEMO.

Novel dual-responsive phenothiazine-based probes for moisture detection and Cu2+ sensing

Development of a fluorescent probe with an ability to detect two different analytes is challenging. Implanting two distinct properties into a single molecule is necessary to attain this dual-responsiveness. In the current study, we prepared two phenothiazine-based probes (probes 1 and 2) with the abilities to detect both moisture in organic solvents and Cu2+ in ethanol (EtOH). The presence of moisture in organic solvents is often detrimental for various applications including preparations of pharmaceuticals and fine chemicals, while high levels of Cu2+ are harmful to human health. Probe 1 exhibited luminescence with different colors in various solvents depending on the solvent polarity. Luminescence of this probe was responsive to a trace amount of moisture present in commonly used organic solvents such as DMSO, DMF, and EtOH. Due to a dramatic change in luminescence intensity and/or color, the probe showed the ability to differentiate water percentages in the organic solvents. The minimum limit of detection (LOD) of this probe for moisture sensing was 0.002 %. Intriguingly, the probe displayed selective sensing of Cu2+ in EtOH among various metal ions. Upon Cu2+ binding, the luminescence color of the probe changed from orange-red to greenish-blue, allowing us to use a ratio-metric approach to obtain a fluorescence-based calculated LOD of 1.0 nM and observed LOD of 0.2 µM. This dual-sensing ability achieved by a single probe (i.e., moisture sensing and Cu2+ detection) was well reproduced even in solid-state probe 1 using cotton and filter paper. Thus, the phenothiazine-cored probe holds significant potential in on-site monitoring of moisture in organic solvents and Cu2+ sensing.