Intrinsic Sensitivity Research Articles

Graphene oxide modified tilted fiber Bragg grating for 10<sup>–12</sup> level heavy metal ion sensing

Graphene oxide (GO) is an ideal label-free sensing material with its super large specific surface area and abundant surface functional groups. Considering its insulating characteristic, the GO is suitable for optics-based heavy metal ion sensing. However, given the large surface tension of water and the hydrophilicity of GO, the agglomeration or wrinkles of GO nanosheets is usually inevitable during coating with aqueous dispersion. This reduces the accessible surface area and surface functional groups of GO, thereby degrading the sensing performance. Here, an ultra-sensitive GO functionalized tilted fiber Bragg grating (TFBG) sensor is designed to detect heavy metal ions in aqueous solutions. Firstly, a strategy of free energy manipulation is employed to avoid the wrinkles and agglomeration of GO nanosheets. In the scenario of aqueous dispersion, the GO nanosheets will wrinkle as the water droplets evaporate and shrink. In contrast, using the lower-surface-tension ethanol as the dispersant and a high-surface-energy substrate processed by oxygen plasma, the dispersion will evenly spread on the substrate instead of forming droplets. When ethanol evaporates, GO nanosheets are attached to the substrate in largest possible area to reduce the free energy of the system, by which a GO film without agglomeration or wrinkles can be obtained. Secondly, the intrinsic sensitivity of TFBG is conducive to the detection of heavy metal ions in water. Mode interference occurs between the cladding mode and the core mode in the TFBG, and the wavelength and intensity of the interference are highly sensitive to the surrounding temperature, stress, and refractive index. Combining the above characteristics, the GO functionalized TFBG is highly sensitive to Pb<sup>2+</sup>, Cd<sup>2+</sup>, and Cu<sup>2+</sup> ions in water. These heavy metal ions are adsorbed by the GO, and thus causing the effective refractive index to increase. The results show that the adsorption of heavy metal ions makes the interference peaks red-shifted in the transmission spectrum. The lowest detection limit for Pb<sup>2+</sup> and Cd<sup>2+</sup> can reach 10<sup>–10</sup> mol/L (ng/L level), and the corresponding sensitivities are 0.426 and 0.385 dB/(nmol·L<sup>–1</sup>) (2.06 and 3.43 dB/(μg·L<sup>–1</sup>)), respectively. These superior sensing performances benefit from the high specific surface area and accessible carbonyl groups of the unfolded GO, and also rely on the excellent intrinsic sensitivity of TFBG. The GO functionalized TFBG sensor has a promising potential application in environment monitoring.

Standardization of molecular techniques for the detection and characterization of intestinal protozoa and other pathogens in humans.

Background: The intrinsic sensitivity limitations of basic parasitological methods, along with the particular biological characteristics of parasites, make these methods ineffective to differentiate morphologically indistinguishable species. Molecular detection and characterization techniques could be used to overcome these problems. The purpose of this work was to standardize molecular polymerase chain reaction (PCR) techniques, described in the literature, for the detection and molecular characterization of intestinal protozoa and other pathogens in humans. Methods: DNA was extracted from human or animal feces, previously washed or cultured in Boeck Drbohlav's Modified Medium. DNA extraction was performed with Machery-Nagel extraction kits. The standardization of the PCR, nested-PCR or RFLP techniques was carried out according to the literature. For each molecular technique performed, the sensitivity of the test was determined based on the minimun quantity required of DNA (sensitivity A) and the minimum quantity of life forms that the test detected (sensitivity B). Results: Sensitivity A was 10 fg for G. duodenalis, 12.5 pg for Entamoeba histolytica or Entamoeba dispar, 50 fg for Cryptosporidium spp., 225 pg for Cyclospora spp. and 800 fg or 8 fg for Blastocystis spp. after performing a 1780 bp PCR or 310 bp nested PCR, respectively. The sensitivity B was 100 cysts for G. duodenalis, 500 cysts for E. histolytica or E. dispar, 1000 oocysts for Cyclospora spp. and 3600 or four vegetatives forms for PCR or nested PCR of Blastocystis spp., respectively. Conclusions: The molecular detection of protozoa and chromist was achieved and the molecular characterization allowed the genotyping of some of the parasites such as Giardia duodenalis, Cryptosporidium spp., and Blastocystis spp. This study summarizes the molecular techniques for epidemiological studies in humans and animals, and helps in the investigation of their transmission sources in countries where intestinal parasites are a public health problem.

Highly bioresistant, hydrophilic and rigidly linked trityl-nitroxide biradicals for cellular high-field dynamic nuclear polarization.

Cellular dynamic nuclear polarization (DNP) has been an effective means of overcoming the intrinsic sensitivity limitations of solid-state nuclear magnetic resonance (ssNMR) spectroscopy, thus enabling atomic-level biomolecular characterization in native environments. Achieving DNP signal enhancement relies on doping biological preparations with biradical polarizing agents (PAs). Unfortunately, PA performance within cells is often limited by their sensitivity to the reductive nature of the cellular lumen. Herein, we report the synthesis and characterization of a highly bioresistant and hydrophilic PA (StaPol-1) comprising the trityl radical OX063 ligated to a gem-diethyl pyrroline nitroxide via a rigid piperazine linker. EPR experiments in the presence of reducing agents such as ascorbate and in HeLa cell lysates demonstrate the reduction resistance of StaPol-1. High DNP enhancements seen in small molecules, proteins and cell lysates at 18.8 T confirm that StaPol-1 is an excellent PA for DNP ssNMR investigations of biomolecular systems at high magnetic fields in reductive environments.

Frozen water NMR lineshape analysis enables absolute polarization quantification.

Typical magnetic resonance experiments are routinely limited by weak signal responses. In some cases, the low intrinsic sensitivity can be alleviated by the implementation of hyperpolarization technologies. Dissolution-dynamic nuclear polarization offers a means of hyperpolarizing small molecules. Hyperpolarized water is employed in several dynamic nuclear polarization studies, and hence accurate and rapid quantification of the 1H polarization level is of utmost importance. The solid-state nuclear magnetic resonance spectrum of water acquired under dissolution-dynamic nuclear polarization conditions has revealed lineshapes which become asymmetric at high levels of 1H polarization, which is an interesting fundamental problem in itself, but also complicates data interpretation and can prevent correct estimations of polarization levels achieved. In previous studies, attempts to simulate the 1H spectral lineshape of water as a function of the 1H polarization led to significant disagreement with the experimental results. Here we propose and demonstrate that such simulations, and therefore polarization quantification, can be implemented accurately, in particular by taking into account the detector dead time during 1H signal acquisition that can lead to severe spectral distortions. Based on these findings, we employed an echo-based radiofrequency pulse sequence to achieve distortion-free 1H spectra of hyperpolarized water, and adequate simulations of these echo-based spectra were implemented to extract the absolute 1H polarization level from the hyperpolarized water signal only, thus alleviating the need for lengthy and insensitive measurements of thermal equilibrium signals.

The effect of catalyst formulation and Rh dispersion on the performance of a CPO fuel processor investigated by operando sampling technique and predictive modelling analysis

The catalytic partial oxidation (CPO) of methane and iso-octane on Rh-coated monoliths is studied in an adiabatic reactor where axial temperature and concentration profiles are collected by a spatially resolved sampling technique. In CH4-CPO, the Rh/MgAl2O4 outperforms the Rh/α-Al2O3 formulation, due to a significant improvement of Rh dispersion. In iso-octane CPO, the beneficial effect of the improved Rh surface is less important due to the intrinsic lower sensitivity of the system. However, a non-negligible impact of Rh dispersion on the extent of hydrocarbon side-products is observed. This factor, together with the lower acidity of the spinel support, contributes to limit the C build-up. Reactor model and kinetic schemes allow to rationalize the measurements and explore the more general effect of Rh specific surface on the key performance indicators of the CPO reformer, that is syngas productivity and hot-spot temperature. Gas-solid diffusion rate makes such indicators strictly fuel-specific.

New Strategies to Promote Macrophage Cholesterol Efflux.

The capacity of macrophages to dispose of cholesterol deposited in the atherosclerotic plaque depends on their ability to activate cholesterol efflux pathways. To develop athero-protective therapies aimed at promoting macrophage cholesterol efflux, cholesterol metabolism in THP-1 monocyte-derived macrophages has been extensively studied, but the intrinsic sensitivity of monocytes and the lack of a standardized procedure to differentiate THP-1 monocytes into macrophages have made it difficult to utilize THP-1 macrophages in the same or similar degree of differentiation across studies. The variability has resulted in lack of understanding of how the differentiation affects cholesterol metabolism, and here we review and investigate the effects of THP-1 differentiation on cholesterol efflux. The degree of THP-1 differentiation was inversely associated with ATP binding cassette A1 (ABCA1) transporter-mediated cholesterol efflux. The differentiation-associated decrease in ABCA1-mediated cholesterol efflux occurred despite an increase in ABCA1 expression. In contrast, DSC1 expression decreased during the differentiation. DSC1 is a negative regulator of the ABCA1-mediated efflux pathway and a DSC1-targeting agent, docetaxel showed high potency and efficacy in promoting ABCA1-mediated cholesterol efflux in THP-1 macrophages. These data suggest that pharmacological targeting of DSC1 may be more effective than increasing ABCA1 expression in promoting macrophage cholesterol efflux. In summary, the comparison of THP-1 macrophage subtypes in varying degrees of differentiation provided new insights into cholesterol metabolism in macrophages and allowed us to identify a viable target DSC1 for the promotion of cholesterol efflux in differentiated macrophages. Docetaxel and other pharmacological strategies targeting DSC1 may hold significant potential for reducing atherogenic cholesterol deposition.

Responsive Sensors of Upconversion Nanoparticles.

Upconversion nanoparticles are a class of luminescent materials that convert longer-wavelength near-infrared photons into visible and ultraviolet emissions. They can respond to various external stimuli, which underpins many opportunities for developing the next generation of sensing technologies. In this perspective, the unique stimuli-responsive properties of upconverting nanoparticles are introduced, and their recent implementations in sensing are summarized. Promising material development strategies for enhancing the key sensing merits, including intrinsic sensitivity, biocompatibility and modality, are identified and discussed. The outlooks on future technological developments, novel sensing concepts, and applications of nanoscale upconversion sensors are provided.

A Microfluidic Cancer-on-Chip Platform Predicts Drug Response Using Organotypic Tumor Slice Culture.

Optimal treatment of cancer requires diagnostic methods to facilitate therapy choice and prevent ineffective treatments. Direct assessment of therapy response in viable tumor specimens could fill this diagnostic gap. Therefore, we designed a microfluidic platform for assessment of patient treatment response using tumor tissue slices under precisely controlled growth conditions. The optimized Cancer-on-Chip (CoC) platform maintained viability and sustained proliferation of breast and prostate tumor slices for 7 days. No major changes in tissue morphology or gene expression patterns were observed within this time frame, suggesting that the CoC system provides a reliable and effective way to probe intrinsic chemotherapeutic sensitivity of tumors. The customized CoC platform accurately predicted cisplatin and apalutamide treatment response in breast and prostate tumor xenograft models, respectively. The culture period for breast cancer could be extended up to 14 days without major changes in tissue morphology and viability. These culture characteristics enable assessment of treatment outcomes and open possibilities for detailed mechanistic studies. SIGNIFICANCE: The Cancer-on-Chip platform with a 6-well plate design incorporating silicon-based microfluidics can enable optimal patient-specific treatment strategies through parallel culture of multiple tumor slices and diagnostic assays using primary tumor material.

When does temperature matter for ecosystem respiration?

The temperature response of ecosystem processes is key to understand and predict impacts of climate change. This is especially true for respiration, given its high temperature sensitivity and major role in the global carbon cycle. However, similar intrinsic temperature sensitivity for respiration does not mean comparable temperature effects across ecosystems and biomes because non-temperature factors can be more important. Here we analyzed soil and sediment respiration data and found that in temperature ranges corresponding to high latitude mean temperatures, absolute respiration rates are more sensitive to non-temperature factors than to projected direct temperature effects. However, at higher temperatures (>20 °C) the direct effect of temperature mediated by temperature sensitivity will likely be more important over changes in non-temperature factors in shaping how respiration change over time. This supports past suggestions that the relatively small projected temperature increase at low (tropical) latitudes may have a large direct impact on absolute respiration. In contrast, absolute respiration rates at high (boreal/arctic) latitudes will likely be more sensitive on the development of the non-temperature factors than on the direct effects of the large projected temperature increase there. Social media abstract. Respiration may be less dependent to changes in temperature at higher than lower latitudes.

Temperature and pH mediate stoichiometric constraints of organically derived soil nutrients.

It remains unclear how warming will affect resource flows during soil organic matter (SOM) decomposition, in part due to uncertainty in how exoenzymes produced by microbes and roots will function. Rising temperatures can enhance the activity of most exoenzymes, but soil pH can impose limitations on their catalytic efficiency. The effects of temperature and pH on enzyme activity are often examined in environmental samples, but purified enzyme kinetics reveal fundamental attributes of enzymes’ intrinsic temperature responses and how relative release of decay‐liberated resources (their flow ratios) can change with environmental conditions. In this paper, we illuminate the principle that fundamental, biochemical limitations on SOM release of C, N, and P during decay, and differential exoenzymes’ responses to the environment, can exert biosphere‐scale significance on the stoichiometry of bioavailable soil resources. To that end, we combined previously published intrinsic temperature sensitivities of two hydrolytic enzymes that release C and N during decay with a novel data set characterizing the kinetics of a P‐releasing enzyme (acid phosphatase) across an ecologically relevant pH gradient. We use these data to estimate potential change in the flow ratios derived from these three enzymes’ activities (C:N, C:P, and N:P) at the global scale by the end of the century, based on temperature projections and soil pH distribution. Our results highlight how the temperature sensitivity of these hydrolytic enzymes and the influence of pH on that sensitivity can govern the relative availability of bioavailable resources derived from these enzymes. The work illuminates the utility of weaving well‐defined kinetic constraints of microbes’ exoenzymes into models that incorporate changing SOM inputs and composition, nutrient availability, and microbial functioning into their efforts to project terrestrial ecosystem functioning in a changing climate.

Sensitivity of soil respiration rate with respect to temperature, moisture and oxygen under freezing and thawing

In alpine environments, the decomposition rate of soil organic carbon (SOC) is controlled by several biotic and abiotic factors, which mostly change simultaneously and often lead to freezing and thawing cycles. However, it is highly uncertain whether the temperature sensitivity of decomposition around the freezing point of water is similar as in higher temperature ranges. In this study, we conducted a full factorial incubation experiment using soil samples from a grassland site in the Tibetan Plateau. A manipulative freeze-thaw cycle was imposed to these soils by continuously changing temperature, from −5 to 10 °C. Additional treatments included 4 levels of soil moisture at 15, 30, 60 and 90% of water-filled pore space (WFPS), and two levels of O2 concentration at 0 and 20%. We fitted the Arrhenius equation into the flux data to estimate the activation energy (Ea) and base flux rate (A) for each treatment level. Then, we predicted the dependence and sensitivity of decomposition rate (k) by implementing the Dual Arrhenius and Michaelis-Menten (DAMM) model using a Bayesian optimization approach. While soil temperature had the strongest control on SOC decomposition rate at all soil moisture and O2 levels, its intrinsic temperature sensitivity (Δk/ΔT) remained nearly constant across the entire temperature range except around 0 °C. We found that Ea was higher in nearly dry or anoxic conditions, suggesting that in these extremes more energy is required for microbial activity to take place. These intrinsic sensitivities revealed that temperature (energy) is the main factor that limits decomposition in cold environments provided that moisture and oxygen are sufficiently available. Intrinsic sensitivities with respect to soil moisture and oxygen concentration were only relevant at very narrow ranges, when soils were almost dry or partially anoxic, and small changes within these narrow ranges may lead to very strong changes in decomposition rates.

Abstract 12178: Photon Counting Computed Tomography for Aortic Valve Calcium Scoring in Contrast- Enhanced Datasets of the Heart

Introduction: The prognostic value of aortic valve calcium in patients prior to TAVR is well established. Its reliable quantification requires a non-contrast CT of the heart (‘true non contrast’, TNC) which in this cohort is not regularly performed at all centers. Photon Counting Detector CT (PCD-CT) permits the routine derivation of ‘virtual non-contrast’ (VNC) series from CT Angiography (CTA) acquisitions due to its intrinsic spectral sensitivity. These might render separate non-contrast acquisitions unnecessary. The aim of our study was to evaluate the feasibility of aortic valve calcium quantification on VNC series derived from CTA scans. Methods: All patients underwent TAVR planning CT on a novel PCD-CT (NAEOTOM Alpha, Siemens Healthineers, Erlangen, Germany) comprising a non-contrast CT of the heart, followed by a CTA of the heart, aorta and iliac arteries. VNC series were derived from CTA acquisitions and two readers independently quantified aortic valve calcium scores and volumes on VNC- and TNC-series. Results: Here, we report initial results of the first 5 of 100 patients. In comparison with those derived from TNC series, calcium scores and volumes derived from VNC series were consistently lower (mean 713 vs. 1110 and 563 vs. 894 mm 3 ) but correlation was excellent for both calcium score and volume (r’s: 0.945 and 0.938, respectively, both p’s=0.01). Conclusions: Our initial patient experience with TAVR planning CT on a novel PCD-CT shows excellent correlation of aortic valve calcium scores and volumes derived from VNC series, as compared with those derived from TNC series. These will most likely be interchangeable, thus rendering non-contrast acquisitions redundant for TAVR planning. Figure 1 Corresponding series of a TAVR patient with high-grade aortic stenosis demonstrating severe aortic valve calcifications on (A) true non-contrast CT (TNT-Score: 3772), (B) CTA for annulus measurement and (C) VNC-reconstructions derived from CTA (VNC-Score: 3099).

Ion-Selective Membrane-Coated Graphene-Hexagonal Boron Nitride Heterostructures for Field-Effect Ion Sensing.

An intrinsic ion sensitivity exceeding the Nernst–Boltzmann limit and an sp2-hybridized carbon structure make graphene a promising channel material for realizing ion-sensitive field-effect transistors with a stable solid–liquid interface under biased conditions in buffered salt solutions. Here, we examine the performance of graphene field-effect transistors coated with ion-selective membranes as a tool to selectively detect changes in concentrations of Ca2+, K+, and Na+ in individual salt solutions as well as in buffered Locke’s solution. Both the shift in the Dirac point and transconductance could be measured as a function of ion concentration with repeatability exceeding 99.5% and reproducibility exceeding 98% over 60 days. However, an enhancement of selectivity, by about an order magnitude or more, was observed using transconductance as the indicator when compared to Dirac voltage, which is the only factor reported to date. Fabricating a hexagonal boron nitride multilayer between graphene and oxide further increased the ion sensitivity and selectivity of transconductance. These findings incite investigating ion sensitivity of transconductance in alternative architectures as well as urge the exploration of graphene transistor arrays for biomedical applications.

Rapid detection of novel coronavirus SARS-CoV-2 by RT-LAMP coupled solid-state nanopores

The current pandemic of COVID-19 caused by SARS-CoV-2 (severe acute respiratory syndrome coronavirus-2) has raised significant public health concerns. Rapid and accurate testing of SARS-CoV-2 is urgently needed for early detection and control of the disease spread. Here, we present an RT-LAMP coupled glass nanopore digital counting method for rapid detection of SARS-CoV-2. We validated and compared two one-pot RT-LAMP assays targeting nucleocapsid (N) and envelop (E) genes. The nucleocapsid assay was adopted due to its quick time to positive and better copy number sensitivity. For qualitative positive/negative classification of a testing sample, we used the glass nanopore to digitally count the RT-LAMP amplicons and benchmarked the event rate with a threshold. Due to its intrinsic single molecule sensitivity, nanopore sensors could capture the amplification dynamics more rapidly (quick time to positive). We validated our RT-LAMP coupled glass nanopore digital counting method for SARS-CoV-2 detection by using both spiked saliva samples and COVID-19 clinical nasopharyngeal swab samples. The results obtained showed excellent agreement with the gold standard RT-PCR assay. With its integration capability, the electronic nanopore digital counting platform has significant potential to provide a rapid, sensitive, and specific point-of-care assay for SARS-CoV-2.

Mismatch Repair Status in Patient-Derived Colorectal Cancer Organoids Does Not Affect Intrinsic Tumor Cell Sensitivity to Systemic Therapy.

Simple SummaryThe large majority of patients with metastatic colorectal cancer (mCRC) receive chemotherapy with or without targeted therapy. Tumors with a deficient DNA mismatch repair (dMMR) system respond poorly to systemic therapy and are associated with poor prognosis. However, it is unclear whether dMMR causes therapy resistance in a tumor cell-intrinsic manner, or whether other mechanisms underlie this association. We address this issue by exposing a panel of MMR-deficient and -proficient Patient-Derived Organoids (PDOs) to a series of clinically relevant drugs. We show that MMR status did not correlate with the response of PDOs to any of the drugs tested. By contrast, the presence of activating mutations in the KRAS and BRAF oncogenes was significantly associated with resistance to chemotherapy and sensitivity to drugs targeting oncogene-activated pathways. We conclude that tumor cell-intrinsic signals link oncogene status, but not MMR status, to variation in therapy response in CRC.DNA mismatch repair deficiency (dMMR) in metastatic colorectal cancer (mCRC) is associated with poor survival and a poor response to systemic treatment. However, it is unclear whether dMMR results in a tumor cell-intrinsic state of treatment resistance, or whether alternative mechanisms play a role. To address this, we generated a cohort of MMR-proficient and -deficient Patient-Derived Organoids (PDOs) and tested their response to commonly used drugs in the treatment of mCRC, including 5-fluorouracil (5-FU), oxaliplatin, SN-38, binimetinib, encorafenib, and cetuximab. MMR status did not correlate with the response of PDOs to any of the drugs tested. In contrast, the presence of activating mutations in the KRAS and BRAF oncogenes was significantly associated with resistance to chemotherapy and sensitivity to drugs targeting oncogene-activated pathways. We conclude that mutant KRAS and BRAF impact the intrinsic sensitivity of tumor cells to chemotherapy and targeted therapy. By contrast, tumor cell-extrinsic mechanisms—for instance signals derived from the microenvironment—must underlie the association of MMR status with therapy response. Future drug screens on rationally chosen cohorts of PDOs have great potential in developing tailored therapies for specific CRC subtypes including, but not restricted to, those defined by BRAF/KRAS and MMR status.

Dose enhancement factor caused by gold nanoparticles: influence of the dosimetric sensitivity and radiation dose assessed by electron spin resonance dosimetry

Gold nanoparticles have been extensively used to increase the sensitivity of radiation dosimeters. In this work, nanocomposites of alanine (Ala), 2-methylalanine (2MA), asparagine (Asn) and monosodium glutamate (MSG) containing gold nanoparticles were prepared. The influence of the mass percentage of gold (0.1% up to 3%), absorbed dose (2 Gy–10 kGy) and the intrinsic sensitivity of these materials on the dose enhancement factor (DEF) were investigated. The prepared nanocomposites were characterized by UV–vis absorption spectroscopy and dynamic light scattering technique. Electron spin resonance spectroscopy was employed to assess the dosimetric response. The results revealed that the gold nanoparticles aggregated in the nanocomposites of MSG and Asn but not in the Ala and 2MA samples. Higher DEFs were observed for materials with lower intrinsic sensitivities (Asn and MSG) and for lower doses of radiation, suggesting that the dosimetric response of the nanocomposite dosimeters is governed by the probability of radical recombination. The higher the radiation dose, gold mass percentage and/or intrinsic sensitivity of the dosimetric material, the higher the production of radiation-induced free-radicals, enhancing the probability of radical recombination and resulting in lower DEFs. These results bring new insights about the use of gold nanoparticles to the construction of more sensitive radiation dosimeters.

Specialized Mechanosensory Epithelial Cells in Mouse Gut Intrinsic Tactile Sensitivity

The gastrointestinal (GI) tract extracts nutrients from ingested meals while protecting the organism from infectious agents frequently present in meals. Consequently, most animals conduct the entire digestive process within the GI tract while keeping the luminal contents entirely outside the body, separated by the tightly sealed GI epithelium. Therefore, like the skin and oral cavity, the GI tract must sense the chemical and physical properties of the its external interface to optimize its function. Specialized sensory enteroendocrine cells (EECs) in GI epithelium interact intimately with luminal contents. A subpopulation of EECs express the mechanically gated ion channel Piezo2 and are developmentally and functionally like the skin's touch sensor- the Merkel cell. We hypothesized that Piezo2+ EECs endow the gut with intrinsic tactile sensitivity. We generated transgenic mouse models with optogenetic activators in EECs and Piezo2 conditional knockouts. We used a range of reference standard and novel techniques from single cells to living animals, including single-cell RNA sequencing and opto-electrophysiology, opto-organ baths with luminal shear forces, and invivo studies that assayed GI transit while manipulating the physical properties of luminal contents. Piezo2+ EECs have transcriptomic features of synaptically connected, mechanosensory epithelial cells. EEC activation by optogenetics and forces led to Piezo2-dependent alterations in colonic propagating contractions driven by intrinsic circuitry, with Piezo2+ EECs detecting the small luminal forces and physical properties of the luminal contents to regulate transit times in the small and large bowel. The GI tract has intrinsic tactile sensitivity that depends on Piezo2+ EECs and allows it to detect luminal forces and physical properties of luminal contents to modulate physiology.

Battery Separator Characterization Strategy via Electron Microscopes

Lithium Ion (Li-ion) batteries store energy and power billions of devices globally, so manufacturers face pressure to ensure high energy density while maintaining safety. Among a battery’s components, a thin separator material prevents physical contact between electrodes while directing ion transport within a cell. Both processes impact cell rate performance, cycle life, and safety [1]. Therefore, an in-depth understanding of separator structure is essential to improving batteries [2-4]. 2D and 3D electron microscopy (EM) effectively characterize separator structure; however, due to intrinsic beam sensitivity to most separators, care is needed to accurately characterize separator structure in its native state. Our work tested 2D scanning EM (SEM) and 3D DualBeamTM (Focused Ion Beam-Scanning Electron Microscope, FIB-SEM) imaging conditions to understand their effect on recovered separator structure.We investigated battery separator structure using a Thermo Scientific ApreoTM SEM and Thermo Scientific DualBeam systems equipped with CryoMAT. The SEM assisted with understanding surface-level electron beam damage in 2D and the DualBeam uncovered 3D structure using FIB milling. In the 2D case, we found the accelerating voltage could warp the separator. Low kV imaging is thus recommended to minimize structure damage.Additionally, in 2D SEM cross-section studies, sample preparation could affect separator structure. We tested an Al2O3-coated composite separator by FIB milling at three different temperatures (Figure 1). We found that cryogenic temperatures (-80° C; i.e., cryo-FIB) achieved the highest quality cross-section (Figure 1). FIB milling at either room temperature (25° C) or ultracold temperature (-180° C) both introduced defects on the microstructure.Finally, given the previous result, we collected 3D separator structure via cryo-FIB. With the 3D image stack, we used Thermo Scientific Avizo™ Software to recover structural parameters: open pore volume fraction, closed pore count and volume fraction, pore connectivity, tortuosity, and permeability. Such information should be useful for understanding battery separator performance. It is expected the strategies discussed in this talk will facilitate separator technology development.

Unshielded magnetocardiography contributes to non-invasively rule-out ischemic heart disease in patients with chest pain of uncertain origin

Abstract Background Out of millions of patients (pts) screened every year for chest pain in emergency departments (EDs) only 10% to 20% are affected by an acute coronary syndrome (ACS) or angina due to ischemic heart disease (IHD). Unless ECG recording and serum markers are clearly positive, most of them must undergo complete clinical workout to exclude IHD prior to discharge. Moreover, since even effort-ECG and echocardiography don't exhaustively rule-out IHD, second-level exams are frequently needed, enhancing hospitalization time and related costs. Magnetocardiography (MCG) is increasingly reported as a sensitive, non-invasive, radiation-free method with high negative predictive value (PV) to rule out IHD. Purpose Retrospective cohort study to evaluate the reliability likelihood of unshielded MCG (uMCG) as a tool for rapid rule-out of IHD. Method Rest uMCG data of 263 patients (pts), all chest-pain free at the moment of MCG scan, were retrospectively analyzed. uMCG was recorded with a 36-channel SQUID system (intrinsic sensitivity 30 fT/√Hz, above 1Hz), for 90 seconds. With proprietary software, 13 MCG parameters were automatically calculated from T-wave magnetic field (T-MF): 5 parameters quantifying the dynamics of the MF extrema and eight, obtained from inverse solution with the equivalent magnetic dipole (EMD) model, quantifying the spatial dynamics of the T-wave effective magnetic vector (T-EMV). Pts were stratified as follows: – Group A, chest pain pts, with IHD confirmed by subsequent SPECT and coronary angiography (mean age 66±11 years, 36.7% female); – Group B, chest pain pts, with IHD excluded by subsequent SPECT or coronary angiography (mean age 59±10 years, 28.8% female); – Group C, healthy control subjects (mean age 25±7 years, 48.6% female). Only 60 pts in each group were randomly selected for the analysis. Non-parametric Kruskal-Wallis tests were used to compare the 13 MCG predictors among groups. Linear discriminant analysis was employed to assess likelihood ratios (LHR) and PVs, considering a mean IHD prevalence of 13% (range 10–20%) in chest-pain pts. Results Positive LHR was 9, while the negative LHR was 0.3, resulting in a helpful reduction of the pre-test probability of IHD. Whereas positive PV ranges from 51 to 70%, hardly confirming IHD diagnosis, negative PV ranges from 92 to 96%, giving confidence for a safe IHD rule-out. Conclusions Automatic analysis of T-MF and of T-EMV dynamics from rest MCG provided impressive negative LHR and PV values that, if confirmed in prospective multicenter trials, would rank MCG as a useful tool for quick and early triage of pts with acute chest pain and still non-diagnostic ECG and enzyme patterns, to rule-out those whose symptoms are of non-ischemic origin. Interestingly, a similar high negative PV has been recently reported using MCG QRS predictors. Thus, even better results could be expected by combining algorithms for automatic assessment of both T-wave and QRS predictors. Funding Acknowledgement Type of funding sources: None.

An alpha II spectrin mutant peptide with unstable scaffold structure and increased sensitivity to calpain cleavage

A spontaneous missense mutation in the alpha II spectrin (αII) gene, replacing a highly conserved arginine 1098 with the glutamine (R1098Q), causes progressive neurodegeneration in heterozygous mutant mice. The molecular mechanism underlying this phenotype is unknown but the accumulation of 150kD αII breakdown products in brains of homozygous mutant embryos suggests an imbalance in the substrate level control of αII cleavage by calpains. This is further supported by in silico simulation predicting unmasked calpain target site and increased spectrin scaffold bending and flexibility of R1098Q mutant peptide. Here, using spectroscopic and in situ enzymatic techniques, we aimed at obtaining direct experimental support for the impact of R1098Q mutation on the αII stability and its propensity for calpain-mediated degradation. Thermal circular dichroism analyses performed on recombinant wildtype and R1098Q mutant αII peptides, composed of spectrin repeat 9–10 revealed that although both had very similar secondary structure contents, thermal stability curve profiles varied and the observed midpoint of the unfolding transition (Tm) was 5.5 °C lower for the R1098Q peptide. Yet, the dynamic light scattering profiles of both peptides closely overlapped, implying the same thermal propensity to aggregate. Calpain digestion of plate-bound αII peptides with and without added calmodulin revealed an enhancement of the R1098Q peptide digestion rate relative to WT control. In summary, these results support the unstable scaffold structure of the R1098Q peptide as contributing to its enhanced intrinsic sensitivity to calpain and suggest physiologic relevance of a proper calpain/spectrin balance in preventing neurodegeneration.