Rapid Signal Research Articles

Fast signaling by glucocorticoids shapes neural representations of behaviors

Glucocorticoids are stress hormones that play central roles in the immediate and slower adaptive responses of the brain and body to new behavioral experience. The exact mechanisms by which the rapid and slow processes underlying glucocorticoid mnemonic effects unfold are under intensive scrutiny. It is possible that glucocorticoids rapidly modify memory representations in the brain by interfering with synaptic functions between inhibitory and excitatory neurons in a timing and context dependent manner. In particular, activity-dependent trans-synaptic messengers appear to have all the necessary attributes to engage in the rapid signaling by glucocorticoids and regulate the brain and behaviors. Novel frameworks for the treatment of stress-related disorders could emerge from a better characterization of the dynamic interplay between the rapid and slow signaling components by glucocorticoids on large-scale brain networks. Here I present some of the exact factors that could help reach this objective.

The Effect of Substrate Stiffness on Elastic Force Transmission in the Epithelial Monolayers over Short Timescales

PurposeThe importance of mechanical forces and microenvironment in guiding cellular behavior has been widely accepted. Together with the extracellular matrix (ECM), epithelial cells form a highly connected mechanical system subjected to various mechanical cues from their environment, such as ECM stiffness, and tensile and compressive forces. ECM stiffness has been linked to many pathologies, including tumor formation. However, our understanding of the effect of ECM stiffness and its heterogeneities on rapid force transduction in multicellular systems has not been fully addressed.MethodsWe used experimental and computational methods. Epithelial cells were cultured on elastic hydrogels with fluorescent nanoparticles. Single cells were moved by a micromanipulator, and epithelium and substrate deformation were recorded. We developed a computational model to replicate our experiments and quantify the force distribution in the epithelium. Our model further enabled simulations with local stiffness gradients.ResultsWe found that substrate stiffness affects the force transduction and the cellular deformation following an external force. Also, our results indicate that the heterogeneities, e.g., gradients, in the stiffness can substantially influence the strain redistribution in the cell monolayers. Furthermore, we found that the cells’ apico-basal elasticity provides a level of mechanical isolation between the apical cell–cell junctions and the basal focal adhesions.ConclusionsOur simulation results show that increased ECM stiffness, e.g., due to a tumor, can mechanically isolate cells and modulate rapid mechanical signaling between cells over distances. Furthermore, the developed model has the potential to facilitate future studies on the interactions between epithelial monolayers and elastic substrates.

The ABI3-ERF1 module mediates ABA-auxin crosstalk to regulate lateral root emergence

Abscisic acid (ABA) is involved in lateral root (LR) development, but how ABA signaling interacts withauxin signaling to regulate LR formation is not well understood. Here, we report that ABA-responsive ERF1 mediates the crosstalk between ABA and auxin signaling to regulate Arabidopsis LR emergence. ABI3 is a negative factor in LR emergence and transcriptionally activates ERF1 by binding to its promoter, and reciprocally, ERF1 activates ABI3, which forms a regulatory loop that enables rapid signal amplification. Notably, ABI3 physically interacts with ERF1, reducing the cis element-binding activities of bothERF1 and ABI3 and thus attenuating the expression of ERF1-/ABI3-regulated genes involved inLR emergence and ABA signaling, such as PIN1, AUX1, ARF7, and ABI5, which may provide a molecular rheostat to avoid overamplification of auxin and ABA signaling. Taken together, our findings identify the role of the ABI3-ERF1 module in mediating crosstalk between ABA and auxin signaling in LR emergence.

4-ATP-modified CNTs@NiO–Fe2O3–Ag SERS filter membrane for rapid in-situ detection of furfural in mineral oil

Evaluating the transformer aging state and detecting multi-aging characteristics in transformer oil with high sensitivity and fast speed has become a key challenge. This study introduces a P–N heterojunction (CNTs@NiO-α-Fe2O3) fabricated through electroless nickel plating and a one-step hydrothermal method. Additionally, silver nanoparticles (AgNPs) with adjustable particle sizes are grown on the surface using a chemical reduction method. To obtain high sensitivity and rapid SERS signal, CNTs@NiO-α-Fe2O3–Ag gel is adsorbed on a disposable needle filter (220 nm) surface, and 4-aminothiophene (4-ATP) is grafted onto the surface of SERS substrate. The minimum detection limit was 0.025 mg/L (EF = 5.22 × 104), and the response time of SERS best signal could be shortened to 3 min. Density functional theory (DFT) calculations reveal that by constructing a P–N heterostructure of NiO–Fe2O3 and assessing the adsorption energies of furfural, acetone, and methanol on the surface of the P–N heterojunction. This SERS strategy has a huge application prospect in the aging diagnosis of oil-paper insulation systems in a transformer.

Differential regulation of BIRC2 and BIRC3 expression by inflammatory cytokines and glucocorticoids in pulmonary epithelial cells.

Roles for the baculoviral inhibitor of apoptosis repeat-containing (BIRC) genes, BIRC2 and BIRC3, may include signaling to the inflammatory transcription factor, nuclear factor-κB (NF-κB) and protection from cell death. However, distinct functions for each BIRC are not well-delineated. Given roles for the epithelium in barrier function and host defence, BIRC2 and BIRC3 expression was characterized in pulmonary epithelial cell lines and primary human bronchial epithelial cells (pHBECs) grown as undifferentiated cells in submersion culture (SC) or as highly differentiated cells at air-liquid interface (ALI). In A549 cells, interleukin-1β (IL1B) and tumor necrosis factor α (TNF) induced BIRC3 mRNA (~20-50-fold), with maximal protein expression from 6-24 h. Similar effects occurred in BEAS-2B and Calu-3 cells, as well as SC and ALI pHBECs. BIRC2 protein was readily detected in unstimulated cells, but was not markedly modulated by IL1B or TNF. Glucocorticoids (dexamethasone, budesonide) modestly increased BIRC3 mRNA and protein, but showed little effect on BIRC2 expression. In A549 cells, BIRC3 mRNA induced by IL1B was unchanged by glucocorticoids and showed supra-additivity with TNF-plus-glucocorticoid. Supra-additivity was also evident for IL1B-plus-budesonide induced-BIRC3 in SC and ALI pHBECs. Using A549 cells, IL1B- and TNF-induced BIRC3 expression, and to a lesser extent, BIRC2, was prevented by NF-κB inhibition. Glucocorticoid-induced BIRC3 expression was prevented by silencing and antagonism of the glucocorticoid receptor. Whereas TNF, but not IL1B, induced degradation of basal BIRC2 and BIRC3 protein, IL1B- and TNF-induced BIRC3 protein remained stable. Differential regulation by cytokines and glucocorticoids shows BIRC2 protein expression to be consistent with roles in rapid signaling events, whereas cytokine-induced BIRC3 may be more important in later effects. While TNF-induced degradation of both BIRCs may restrict their activity, cytokine-enhanced BIRC3 expression could prime for its function. Finally, shielding from glucocorticoid repression, or further enhancement by glucocorticoid, may indicate a key protective role for BIRC3.

Cost-effective selective deuteration of aromatic amino acid residues produces long-lived solution 1H NMR magnetization in proteins

Solution NMR studies of large proteins are hampered by rapid signal decay due to short-range dipolar 1H–1H and 1H–13C interactions. These are attenuated by rapid rotation in methyl groups and by deuteration (2H), so selective 1H,13C-isotope labelling of methyl groups in otherwise perdeuterated proteins, combined with methyl transverse relaxation optimized spectroscopy (methyl-TROSY), is now standard for solution NMR of large protein systems > 25 kDa. For non-methyl positions, long-lived magnetization can be introduced as isolated 1H–12C groups. We have developed a cost-effective chemical synthesis for producing selectively deuterated phenylpyruvate and hydroxyphenylpyruvate. Feeding these amino acid precursors to E. coli in D2O, along with selectively deuterated anthranilate and unlabeled histidine, results in isolated and long-lived 1H magnetization in the aromatic rings of Phe (HD, HZ), Tyr (HD), Trp (HH2, HE3) and His (HD2 and HE1). We are additionally able to obtain stereoselective deuteration of Asp, Asn, and Lys amino acid residues using unlabeled glucose and fumarate as carbon sources and oxalate and malonate as metabolic inhibitors. Combining these approaches produces isolated 1H–12C groups in Phe, Tyr, Trp, His, Asp, Asn, and Lys in a perdeuterated background, which is compatible with standard 1H–13C labeling of methyl groups in Ala, Ile, Leu, Val, Thr, Met. We show that isotope labeling of Ala is improved using the transaminase inhibitor L-cycloserine, and labeling of Thr is improved through addition of Cys and Met, which are known inhibitors of homoserine dehydrogenase. We demonstrate the creation of long-lived 1H NMR signals in most amino acid residues using our model system, the WW domain of human Pin1, as well as the bacterial outer membrane protein PagP.

Temporal Tracking of Insulin Action on the Cell Surface of Proteins at a Resolution of Ten Seconds.

The ligand-receptor signaling occurring on the cell surface governs cell growth, proliferation, and survival via rapidly triggering a cascade of events. Here, we for the first time report an in situ perturbation-free and rapid surface proteomic profiling at a temporal resolution of ten seconds. By this innovation, about 1022 cell surface-associated proteins were reproducibly identified and quantified. It is noteworthy that, upon a model ligand insulin stimulus, a few rapid-responding proteins at 10 s to 2 min were identified, e.g., CNNM3. Moreover, temporal response patterns were established for the members of GLUT4 storage vesicles (GSVs; responsible for glucose transportation) and confirmed with five known GSV proteins. This pattern was then exploited to uncover seven new regulatory proteins (LDLR, HFE, ECE1, MRC2, CORO1C, CPD, and BST2). Collectively, we showed a powerful surface proteomic tool to decipher rapid signaling of cell-surface proteins and to uncover new subunits involved in rapidly trafficking vesicles.

Electrochemical characterization of 17β‐estradiol with fast‐scan cyclic voltammetry

AbstractWe have developed a sensitive and stable electrochemical method for 17β‐estradiol (E2) detection using fast‐scan cyclic voltammetry (FSCV). Recently, E2 was proposed to function as a rapid synaptocrine signaling molecule in the brain; however, methods to directly monitor subsecond fluctuations in E2 are currently unavailable, limiting our understanding of the dynamics and mechanism of rapid E2 release. FSCV at carbon‐fiber microelectrodes enables subsecond detection of electroactive neurochemicals directly in tissues like the brain. Here, we have electrochemically characterized E2 using FSCV for use in a tissue matrix. The limit of detection of E2 is 31.2±2.5 nM with FSCV, which will enable low nanomolar fluctuations in extracellular E2 to be monitored with hundred millisecond temporal resolution. We also identify specific parameters for waveform modification to improve future detection. This method will significantly improve E2 sensing capabilities and will have far‐reaching impacts on improving our understanding of dynamic E2 signaling in the brain.

Basic study on calibration using waveform variation in space charge measurement by pulsed electroacoustic method

AbstractThe pulsed electroacoustic method determines the distribution of space charge in an insulation system by applying a pulse voltage and detecting the acoustic response. It estimates the location and magnitude of charges from time‐domain signals. For quantitative analysis, a bias voltage is applied to a specimen with no space charge, and the acoustic signal generated from the electrodes only is used as a reference waveform for calibration. However, a full‐scale, extra‐high voltage insulation system often already contains space charges in it. Therefore, a method of acquiring a reference signal under such a condition was studied. We studied methods to compare voltage variations and response signal variations. We proposed a method that measures the signal difference at different voltages. In addition, we proposed a method of correlating voltage and signal variations that may allo for more rapid signal acquisition. The feasibility was proved using an 11 mm‐thick mimic polyethylene specimen.

CRISPR/deadCas9-based high-throughput gene doping analysis (HiGDA): A proof of concept for exogenous human erythropoietin gene doping detection.

A genetic approach targeted toward improving athletic performance is called gene doping and is prohibited by the World Anti-Doping Agency. Currently, the clustered regularly interspaced short palindromic repeats-associated protein (Cas)-related assays have been utilized to detect genetic deficiencies or mutations. Among the Cas proteins, deadCas9 (dCas9), a nuclease-deficient mutant of Cas9, acts as a DNA binding protein with a target-specific single guide RNA. On the basis of the principles, we developed a dCas9-based high-throughput gene doping analysis for exogenous gene detection. The assay comprises two distinctive dCas9s, a magnetic bead immobilized capture dCas9 for exogenous gene isolation and a biotinylated dCas9 with streptavidin-polyHRP that enables rapid signal amplification. For efficient biotin labeling via maleimide-thiol chemistry, two cysteine residues of dCas9 were structurally validated, and the Cys574 residue was identified as an essential labeling site. As a result, we succeeded in detecting the target gene in a concentration as low as 12.3fM (7.41×105 copies) and up to 10nM (6.07×1011 copies) in a whole blood sample within 1h with HiGDA. Assuming an exogenous gene transfer scenario, we added a direct blood amplification step to establish a rapid analytical procedure while detecting target genes with high sensitivity. Finally, we detected the exogenous human erythropoietin gene at concentrations as low as 2.5 copies within 90min in 5μL of the blood sample. Herein, we propose that HiGDA is a very fast, highly sensitive, and practical detection method for actual doping field in the future.

Long-Term Health Monitoring Data Processing on Post-Tensioned Concrete Box-Girder Bridge by Wavelet-Based

The concrete box-girder bridge is designed to have a long service life of around 100 years. To ensure safety and performance degradation during long service life, a Structural Health Monitoring System (SHMS) has been implemented in the box-girder bridge. SHMS can reliably assess structural response due to real-time applied loads, detect anomaly activities and locate the structural damage in the structure. Several sensors have been implemented in the bridge to continuously record the behavior of the bridge in all environmental conditions. Due to real-time natural conditions, false alarms occur frequently in SHM due to the disruption of noises and lead to misunderstanding of who is evaluating. Nevertheless, numerous SHM data that have been collected make it complicated to determine the anomaly of the structures. Therefore, it required signal processing to maximize the potentialities of the massive SHM data, as well as the efficiency of the time work. In this study, wavelet transformation, a rapid and unsupervised signal processing approach, was used to analyze the huge signal data by removing noise, and separating different signal sources as well. Further, with time-frequency analysis and multi-resolution capabilities, the transformation of wavelet is a promising tool for analyzing long-term SHM data. The suggested approach is shown by using long-term strain data from a 40 m concrete box-girder bridge in 24h. The results showed that after the denoising process, the highest discrepancy between the reconstructed and original strain signal is 2.73 μƐ and lost their energy less than 1%. Hence, the strain gauge sensor was successfully able to eliminate the noise through wavelet technology.

JAK/STAT Signaling and Cervical Cancer: From the Cell Surface to the Nucleus.

The Janus kinase (JAK)/signal transducer and activator of transcription (STAT) signaling pathway constitutes a rapid signaling module from the cell surface to the nucleus, and activates different cellular responses, such as proliferation, survival, migration, invasion, and inflammation. When the JAK/STAT pathway is altered, it contributes to cancer progression and metastasis. STAT proteins play a central role in developing cervical cancer, and inhibiting the JAK/STAT signaling may be necessary to induce tumor cell death. Several cancers show continuous activation of different STATs, including cervical cancer. The constitutive activation of STAT proteins is associated with a poor prognosis and overall survival. The human papillomavirus (HPV) oncoproteins E6 and E7 play an essential role in cervical cancer progression, and they activate the JAK/STAT pathway and other signals that induce proliferation, survival, and migration of cancer cells. Moreover, there is a crosstalk between the JAK/STAT signaling cascade with other signaling pathways, where a plethora of different proteins activate to induce gene transcription and cell responses that contribute to tumor growth. Therefore, inhibition of the JAK/STAT pathway shows promise as a new target in cancer treatment. In this review, we discuss the role of the JAK/STAT pathway components and the role of the HPV oncoproteins associated with cellular malignancy through the JAK/STAT proteins and other signaling pathways to induce tumor growth.

Different evolutionary patterns of TIR1/AFBs and AUX/IAAs and their implications for the morphogenesis of land plants

BackgroundThe plant hormone auxin is widely involved in plant growth, development, and morphogenesis, and the TIR1/AFB and AUX/IAA proteins are closely linked to rapid auxin response and signal transmission. However, their evolutionary history, historical patterns of expansion and contraction, and changes in interaction relationships are still unknown.ResultsHere, we analyzed the gene duplications, interactions, and expression patterns of TIR1/AFBs and AUX/IAAs to understand their underlying mechanisms of evolution. The ratios of TIR1/AFBs to AUX/IAAs range from 4:2 in Physcomitrium patens to 6:29 in Arabidopsis thaliana and 3:16 in Fragaria vesca. Whole-genome duplication (WGD) and tandem duplication have contributed to the expansion of the AUX/IAA gene family, but numerous TIR1/AFB gene duplicates were lost after WGD. We further analyzed the expression profiles of TIR1/AFBs and AUX/IAAs in different tissue parts of Physcomitrium patens, Selaginella moellendorffii, Arabidopsis thaliana and Fragaria vesca, and found that TIR1/AFBs and AUX/IAAs were highly expressed in all tissues in P. patens, S. moellendorffii. In A. thaliana and F. vesca, TIR1/AFBs maintained the same expression pattern as the ancient plants with high expression in all tissue parts, while AUX/IAAs appeared tissue-specific expression. In F. vesca, 11 AUX/IAAs interacted with TIR1/AFBs with different interaction strengths, and the functional specificity of AUX/IAAs was related to their ability to bind TIR1/AFBs, thus promoting the development of specific higher plant organs. Verification of the interactions among TIR1/AFBs and AUX/IAAs in Marchantia polymorpha and F. vesca also showed that the regulation of AUX/IAA members by TIR1/AFBs became more refined over the course of plant evolution.ConclusionsOur results indicate that specific interactions and specific gene expression patterns both contributed to the functional diversification of TIR1/AFBs and AUX/IAAs.

Sensitive Fluorescent Determination of Silver Ion and Glutathione in Human Serum Using Polydopamine Nanodots as the Probe

Glutathione (GSH)-switched fluorescent assays have received attention due to their rapid signal changes of fluorescent nanoprobes. Small polydopamine nanodots (PDs) exhibit remarkable photoluminescent properties. In this work, fluorescent PDs were synthesized through oxidation using dopamine as precursor. The prepared PDs had rich surface functionalities and exhibited good optical properties with maximum emission using 330 nm excitation. The fluorescence was quenched by Ag+ through the interaction with catechols in the PDs. After the addition of GSH, Ag was bound with sulfhydryl groups in GSH and the fluorescence of the PDs was restored. Therefore, a fluorescence turn on-off-on strategy was constructed for Ag+ and GSH using the synthesized PDs as the probe. The PDs showed good performance for Ag+ from 0.01 to 1 μM, with a limit of detection (LOD) equal to 2.82 nM. The system also exhibited a linear relationship for GSH from 0.01 to 0.5 μM with a LOD equal to 1.93 nM. GSH was determined in human serum to demonstrate a practical application of the assay. This work expands the application of PDs in biological analysis.

The G protein-coupled oestrogen receptor GPER in health and disease: an update.

Oestrogens and their receptors contribute broadly to physiology and diseases. In premenopausal women, endogenous oestrogens protect against cardiovascular, metabolic and neurological diseases and are involved in hormone-sensitive cancers such as breast cancer. Oestrogens and oestrogen mimetics mediate their effects via the cytosolic and nuclear receptors oestrogen receptor-α(ERα) and oestrogen receptor-β (ERβ) and membrane subpopulations as well as the 7-transmembrane G protein-coupled oestrogen receptor (GPER). GPER, which dates back more than 450 million years in evolution, mediates both rapid signalling and transcriptional regulation. Oestrogen mimetics (such as phytooestrogens and xenooestrogens including endocrine disruptors) and licensed drugs such as selective oestrogen receptor modulators(SERMs) and downregulators(SERDs) also modulate oestrogen receptor activity in both health and disease. Following up on our previous Review of 2011, we herein summarize the progress made in the field of GPER research over the past decade. We will review molecular, cellular and pharmacological aspects of GPER signalling and function, its contribution to physiology, health and disease, and the potential of GPER to serve as a therapeutic target and prognostic indicator of numerous diseases. We also discuss the first clinical trial evaluating a GPER-selective drug and the opportunity of repurposing licensed drugs for the targeting of GPER in clinical medicine.

Ten-kilohertz two-photon microscopy imaging of single-cell dendritic activity and hemodynamics in vivo.

The studying of rapid neuronal signaling across large spatial scales in intact, living brains requires both high temporal resolution and versatility of the measurement device. We introduce a high-speed two-photon microscope based on a custom-built acousto-optic deflector (AOD). This microscope has a maximum line scan frequency of 400kHz and a maximum frame rate of 10,000 frames per second (fps) at . For stepwise magnification from population view to subcellular view with high spatial and temporal resolution, we combined the AOD with resonance-galvo (RS) scanning. With this combinatorial device that supports both large-view navigation and small-view high-speed imaging, we measured dendritic calcium propagation velocity and the velocity of single red blood cells (RBCs). We measured dendritic calcium propagation velocity ( ) in OGB-1-labeled single cortical neurons in mice in vivo. To benchmark the spatial precision and detection sensitivity of measurement in vivo, we also visualized the trajectories of single RBCs and found that their movement speed follows Poiseuille's law of laminar flow. This proof-of-concept methodological development shows that the combination of AOD and RS scanning two-photon microscopy provides both versatility and precision for quantitative analysis of single neuronal activities and hemodynamics in vivo.

Multicolor flow cytometry in clinical samples for platelet signaling assessment

BackgroundAvailability of multichannel cytometers and specific commercial antibodies makes flow cytometry a new option to simultaneously assess multiple intracellular platelet signaling pathways for clinical purposes, in small volume of blood or low platelet count. ObjectivesTo describe a multicolor flow cytometry with fluorescent barcoding technique for screening signaling pathways downstream membrane receptors of major platelet agonists (adenosine diphosphate, thrombin, thromboxane, and collagen). MethodsBy comparison with immunoblotting, we first selected the target phosphoproteins, AKT, P38MAPK, LIMK, and SPL76; the times of stimulation; and phosphoflow barcoding conditions. We then performed a clinical study on whole blood of patients without evidence of blood platelet disorder on standard biological screening, consulting for trivial or occasionally provoked bleeds without familial antecedent (bleeding of unknown origin, n = 23) or type-1 von Willebrand disease (n = 9). In addition, we included a small group of patients with definite platelet disorders (Glanzmann thrombasthenia, δ-storage pool deficiency, and immune glycoprotein VI–related disease with granule secretion defect). ResultsThe range, kinetics, and distribution of fluorescence intensity were established for each agonist-target protein combination. Principal component analysis indicates a correlation in response to a target phosphoprotein (AKT and P38MAPK) to different agonists but no correlation in the response of different target phosphoproteins to the same agonist. The heterogeneity of individual responses in the whole population displayed was analyzed using clustering algorithm. Patients with platelet storage pool deficiency were positioned as lowest responders on the heatmap. ConclusionIn complement of functional tests, this study introduces a new approach for rapid platelet signaling profiling in clinical practice.

Thermoluminescence properties of TiO2/Cu/TiO2 multilayer thin films fabricated by (RF/DC) sputtering for radiation dosimetry

The most popular TLDs are composed of phosphors, ceramics, and crystals. Despite having a high light output per unit of absorbed dose, they suffer from rapid signal fading, light scattering, saturation at higher doses, and difficulties annealing for re-use. Here, we report the characteristics of a novel TiO2/Cu/TiO2 multilayer thin film dosimeter fabricated using radio-frequency (RF) and direct current (DC) sputtering methods with a faster deposition rate, high purity, and uniformity. The multilayer sequence was set to TiO2 (150 nm)/Cu (x)/TiO2 (50 nm), at varying Cu thicknesses (x = 5, 10, 25, 40, and 50 nm). The structure, morphology, elemental composition, and optical properties of the films were characterized by the XRD, FESEM, EDX, and UV–Vis, respectively. Next, the best TiO2 multilayer films with 5 nm Cu were tested for morphological, structural, and optical properties at 600 and 1200 mGy of x-ray radiation. The dosimetric properties of samples irradiated with X-ray doses ranging from 100 to 1500 mGy were then investigated. The best annealing time, reproducibility, fading, low minimum detectable dose as well as the effect of different heating rates on the TL glow curve have all been studied. Our results indicate that the films are amorphous, porous with spherically shaped nanoparticles, have a uniform distribution of elements, and are transparent in the visible region. The optimum TL intensity was found at TiO2 (445 nm)/14.2 nm Cu/TiO2 (130 nm) sequence. The dose-response has a good linear correlation to x-ray radiation with a sensitivity that is 0.444 times less than that of TLD-100 sensitivity within the dose range 100–1500 mGy. Upon irradiation, the band gap decreases, crystallinity increases, pores increase, films become rougher, and the absorption band edge extends to higher wavelengths. The single and multilayer TiO2 films have high-intensity peaks at 270 and 274 ∘C, respectively, which is typical of TL glow curve peaks used in TLD applications. There were no significant changes in the TL reading after six times of reuse, and the fading was observed at 49.57% after 15 days of irradiation. The findings indicate that TiO2/Cu/TiO2 multilayer films are appropriate for thermoluminescence dosimetric applications.

Enzymatic Catalysis in Size and Volume Dual-Confined Space of Integrated Nanochannel-Electrodes Chip for Enhanced Impedance Detection of Salmonella.

Nanochannel-based confinement effect is a fascinating signal transduction strategy for high-performance sensing, but only size confinement is focused on while other confinement effects are unexplored. Here, a highly integrated nanochannel-electrodes chip (INEC) is created and a size/volume-dual-confinement enzyme catalysis model for rapid and sensitive bacteria detection is developed. The INEC, by directly sandwiching a nanochannel chip (60µm in thickness) in nanoporous gold layers, creates a micro-droplet-based confinement electrochemical cell (CEC). The size confinement of nanochannel promotes the urease catalysis efficiency to generate more ions, while the volume confinement of CEC significantly enriches ions by restricting diffusion. As a result, the INEC-based dual-confinement effects benefit a synergetic enhancement of the catalytic signal. A 11-times ion-strength-based impedance response is obtained within just 1min when compared to the relevant open system. Combining this novel nanoconfinement effects with nanofiltration of INEC, a separation/signal amplification-integrated sensing strategy is further developed for Salmonella typhimurium detection. The biosensor realizes facile, rapid (<20min), and specific signal readout with a detection limit of 9 CFU mL-1 in culturing solution, superior to most reports. This work may create a new paradigm for studying nanoconfined processes and contribute a new signal transduction technique for trace analysis application.

Hypothesis paper: the development of a regulatory layer in P2B autoinhibited Ca2+-ATPases may have facilitated plant terrestrialization and animal multicellularization

ABSTRACT With the appearance of plants and animals, new challenges emerged. These multicellular eukaryotes had to solve for example the difficulties of multifaceted communication between cells and adaptation to new habitats. In this paper, we are looking for one piece of the puzzle that made the development of complex multicellular eukaryotes possible with a focus on regulation of P2B autoinhibited Ca2+-ATPases. P2B ATPases pump Ca2+ out of the cytosol at the expense of ATP hydrolysis, and thereby maintain a steep gradient between the extra- and intracytosolic compartments which is utilized for Ca2+-mediated rapid cell signaling. The activity of these enzymes is regulated by a calmodulin (CaM)-responsive autoinhibitory region, which can be located in either termini of the protein, at the C-terminus in animals and at the N-terminus in plants. When the cytoplasmic Ca2+ level reaches a threshold, the CaM/Ca2+ complex binds to a calmodulin-binding domain (CaMBD) in the autoinhibitor, which leads to the upregulation of pump activity. In animals, protein activity is also controlled by acidic phospholipids that bind to a cytosolic portion of the pump. Here, we analyze the appearance of CaMBDs and the phospholipid-activating sequence and show that their evolution in animals and plants was independent. Furthermore, we hypothesize that different causes may have initiated the appearance of these regulatory layers: in animals, it is linked to the appearance of multicellularity, while in plants it co-occurs with their water-to-land transition.