NeuroSeg-III: efficient neuron segmentation in two-photon Ca2+ imaging data using self-supervised learning.

Two-photon Ca2+ imaging technology increasingly plays an essential role in neuroscience research. However, the requirement for extensive professional annotation poses a significant challenge to improving the performance of neuron segmentation models. Here, we present NeuroSeg-III, an innovative self-supervised learning approach specifically designed to achieve fast and precise segmentation of neurons in imaging data. This approach consists of two modules: a self-supervised pre-training network and a segmentation network. After pre-training the encoder of the segmentation network via a self-supervised learning method without any annotated data, we only need to fine-tune the segmentation network with a small amount of annotated data. The segmentation network is designed with YOLOv8s, FasterNet, efficient multi-scale attention mechanism (EMA), and bi-directional feature pyramid network (BiFPN), which enhanced the model's segmentation accuracy while reducing the computational cost and parameters. The generalization of our approach was validated across different Ca2+ indicators and scales of imaging data. Significantly, the proposed neuron segmentation approach exhibits exceptional speed and accuracy, surpassing the current state-of-the-art benchmarks when evaluated using a publicly available dataset. The results underscore the effectiveness of NeuroSeg-III, with employing an efficient training strategy tailored for two-photon Ca2+ imaging data and delivering remarkable precision in neuron segmentation.

Hypothalamic supramammillary neurons that project to the medial septum modulate wakefulness in mice.

The hypothalamic supramammillary nucleus (SuM) plays a crucial role in controlling wakefulness, but the downstream target regions participating in this control process remain unknown. Here, using circuit-specific fiber photometry and single-neuron electrophysiology together with electroencephalogram, electromyogram and behavioral recordings, we find that approximately half of SuM neurons that project to the medial septum (MS) are wake-active. Optogenetic stimulation of axonal terminals of SuM-MS projection induces a rapid and reliable transition to wakefulness from non-rapid-eye movement or rapid-eye movement sleep, and chemogenetic activation of SuMMS projecting neurons significantly increases wakefulness time and prolongs latency to sleep. Consistently, chemogenetically inhibiting these neurons significantly reduces wakefulness time and latency to sleep. Therefore, these results identify the MS as a functional downstream target of SuM and provide evidence for the modulation of wakefulness by this hypothalamic-septal projection.

Holistic bursting cells store long-term memory in auditory cortex.

The sensory neocortex has been suggested to be a substrate for long-term memory storage, yet which exact single cells could be specific candidates underlying such long-term memory storage remained neither known nor visible for over a century. Here, using a combination of day-by-day two-photon Ca2+ imaging and targeted single-cell loose-patch recording in an auditory associative learning paradigm with composite sounds in male mice, we reveal sparsely distributed neurons in layer 2/3 of auditory cortex emerged step-wise from quiescence into bursting mode, which then invariably expressed holistic information of the learned composite sounds, referred to as holistic bursting (HB) cells. Notably, it was not shuffled populations but the same sparse HB cells that embodied the behavioral relevance of the learned composite sounds, pinpointing HB cells as physiologically-defined single-cell candidates of an engram underlying long-term memory storage in auditory cortex.

The maximal contrast-enhanced range of CT for differentiating the WHO pathological subtypes and risk subgroups of thymic epithelial tumors.

OBJECTIVE: To explore the value of maximal contrast-enhanced (CEmax) range using contrast-enhanced CT (CECT) imaging in differentiating the pathological subtypes and risk subgroups of thymic epithelial tumors (TETs). METHODS: The pre-treatment-CECT images of 319 TET patients from May 2012 to November 2021 were analyzed retrospectively. The CEmax was defined as the maximum difference between the CT value of the solid tumor on pre-contrast and contrast-enhanced images. The mean CEmax value was calculated at three different tumor levels. RESULTS: There was a significant difference in the CEmax among the eight main pathological subtypes [types A, AB, B1, B2, and B3 thymoma, thymic carcinoma (TC), low-grade neuroendocrine tumor (NET) and high-grade NET] (p < 0.001). Among the eight subtypes, the CEmax values of types A, AB, and low-risk NET were higher than those of the other subtypes (all p < 0.001), and there was no difference among types B1-B3 and high-risk NET (all p > 0.05). There was no difference for CEmax values between NET and TC (p = 0.491). For the risk subgroups, the CEmax of TC (including NET) was 35.35 ± 11.41 HU, which was lower than that of low-risk thymoma (A and AB) (57.73±21.24 HU) (P < 0.001) and was higher than that of high-risk thymoma (B1-B3) (27.37±8.27 HU) (P < 0.001). The CEmax cut-off values were 38.5 HU and 30.5 HU respectively (AUC: 0.829 and 0.712; accuracy, 72.4% and 67.7%). CONCLUSION: The tumor CEmax on CECT helps differentiate the pathological subtypes and risk subgroups of TETs. ADVANCES IN KNOWLEDGE: In this study, an improved simplified risk grouping method was proposed based on the traditional (2004 edition) simplified risk grouping method for TETs. If Type B1 thymoma is classified as high-risk, radiologists using this improved method may improve the accuracy in differentiating risk level of TETs compared with the traditional method.

Spatial multi-omics revealed the impact of tumor ecosystem heterogeneity on immunotherapy efficacy in patients with advanced non-small cell lung cancer treated with bispecific antibody.

BACKGROUND: Immunotherapy for malignant tumors has made great progress, but many patients do not benefit from it. The complex intratumoral heterogeneity (ITH) hindered the in-depth exploration of immunotherapy. Conventional bulk sequencing has masked intratumor complexity, preventing a more detailed discovery of the impact of ITH on treatment efficacy. Hence, we initiated this study to explore ITH at the multi-omics spatial level and to seek prognostic biomarkers of immunotherapy efficacy considering the presence of ITH. METHODS: Using the segmentation strategy of digital spatial profiling (DSP), we obtained differential information on tumor and stromal regions at the proteomic and transcriptomic levels. Based on the consideration of ITH, signatures constructed by candidate proteins in different regions were used to predict the efficacy of immunotherapy. RESULTS: Eighteen patients treated with a bispecific antibody (bsAb)-KN046 were enrolled in this study. The tumor and stromal areas of the same samples exhibited distinct features. Signatures consisting of 11 and 18 differentially expressed DSP markers from the tumor and stromal areas, respectively, were associated with treatment response. Furthermore, the spatially resolved signature identified from the stromal areas showed greater predictive power for bsAb immunotherapy response (area under the curve=0.838). Subsequently, our stromal signature was validated in an independent cohort of patients with non-small cell lung cancer undergoing immunotherapy. CONCLUSION: We deciphered ITH at the spatial level and demonstrated for the first time that genetic information in the stromal region can better predict the efficacy of bsAb treatment. TRIAL REGISTRATION NUMBER: NCT03838848.

In Situ Adjustable Nanogaps and In-Plane Break Junctions.

The ability to precisely regulate the size of a nanogap is essential for establishing high-yield molecular junctions, and it is crucial for the control of optical signals in extreme optics. Although remarkable strategies for the fabrication of nanogaps are proposed, wafer-compatible nanogaps with freely adjustable gap sizes are not yet available. Herein, two approaches for constructing in situ adjustable metal gaps are proposed which allow Ångstrom modulation resolution by employing either a lateral expandable piezoelectric sheet or a stretchable membrane. These in situ adjustable nanogaps are further developed into in-plane molecular break junctions, in which the gaps can be repeatedly closed and opened thousands of times with self-assembled molecules. The conductance of the single 1,4-benzenediamine (BDA) and the BDA molecular dimer is successfully determined using the proposed strategy. The measured conductance agreeing well with the data by employing another well-established scanning tunneling microscopy break junction technique provides insight into the formation of molecule dimer via hydrogen bond at single molecule level. The wafer-compatible nanogaps and in-plane dynamical break-junctions provide a potential approach to fabricate highly compacted devices using a single molecule as a building block and supply a promising in-plane technique to address the dynamical properties of single molecules.

Graphene oxide promotes V-Cu-Ce-ZSM-5 to catalyze SO2 and NO at low temperature: performance and mechanism.

A catalyst (V-Cu-Ce-ZSM-5) was explored to simultaneously remove the SO2 and NOx from flue gas by use of the ZSM-5 molecular sieve as the carrier, V and Cu as the active components, and Ce as the additive in low temperature of 150 °C. The performance of V-Cu-Ce-ZSM-5 was evaluated for the oxidation of NO and SO2 before and after the addition of graphene oxide (GO). The results showed that V-Cu-Ce-ZSM-5@GO0.5 had the best performance at a reaction temperature of 150 °C, and the oxidation efficiency of SO2 and NO was 94.60% and 83.64%, respectively. The multiple structural characterizations (BET, SEM, Raman, XRD, and XPS) revealed that the loading of V and Cu with the additive Ce expanded the specific surface area and pore volume of ZSM-5, provided more adsorption sites for SO2 and NO, and had good desulfurization and denitration activity. The addition of GO further improved the dispersibility of active components and auxiliaries, increased the number of active sites in the reaction process, and significantly improved catalytic activity.

Contrast-enhanced CT-based radiomics model for differentiating risk subgroups of thymic epithelial tumors.

BACKGROUND: To validate a contrast-enhanced CT (CECT)-based radiomics model (RM) for differentiating various risk subgroups of thymic epithelial tumors (TETs). METHODS: A retrospective study was performed on 164 patients with TETs who underwent CECT scans before treatment. A total of 130 patients (approximately 79%, from 2012 to 2018) were designated as the training set, and 34 patients (approximately 21%, from 2019 to 2021) were designated as the testing set. The analysis of variance and least absolute shrinkage and selection operator algorithm methods were used to select the radiomics features. A logistic regression classifier was constructed to identify various subgroups of TETs. The predictive performance of RMs was evaluated based on receiver operating characteristic (ROC) curve analyses. RESULTS: Two RMs included 16 and 13 radiomics features to identify three risk subgroups of traditional risk grouping [low-risk thymomas (LRT: Types A, AB and B1), high-risk thymomas (HRT: Types B2 and B3), thymic carcinoma (TC)] and improved risk grouping [LRT* (Types A and AB), HRT* (Types B1, B2 and B3), TC], respectively. For traditional risk grouping, the areas under the ROC curves (AUCs) of LRT, HRT, and TC were 0.795, 0.851, and 0.860, respectively, the accuracy was 0.65 in the training set, the AUCs were 0.621, 0.754, and 0.500, respectively, and the accuracy was 0.47 in the testing set. For improved risk grouping, the AUCs of LRT*, HRT*, and TC were 0.855, 0.862, and 0.869, respectively, and the accuracy was 0.72 in the training set; the AUCs were 0.778, 0.716, and 0.879, respectively, and the accuracy was 0.62 in the testing set. CONCLUSIONS: CECT-based RMs help to differentiate three risk subgroups of TETs, and RM established according to improved risk grouping performed better than traditional risk grouping.

Prestin-Mediated Frequency Selectivity Does not Cover Ultrahigh Frequencies in Mice.

In mammals, the piezoelectric protein, Prestin, endows the outer hair cells (OHCs) with electromotility (eM), which confers the capacity to change cellular length in response to alterations in membrane potential. Together with basilar membrane resonance and possible stereociliary motility, Prestin-based OHC eM lays the foundation for enhancing cochlear sensitivity and frequency selectivity. However, it remains debatable whether Prestin contributes to ultrahigh-frequency hearing due to the intrinsic nature of the cell's low-pass features. The low-pass property of mouse OHC eM is based on the finding that eM magnitude dissipates within the frequency bandwidth of human speech. In this study, we examined the role of Prestin in sensing broad-range frequencies (4-80 kHz) in mice that use ultrasonic hearing and vocalization (to >100 kHz) for social communication. The audiometric measurements in mice showed that ablation of Prestin did not abolish hearing at frequencies >40 kHz. Acoustic associative behavior tests confirmed that Prestin-knockout mice can learn ultrahigh-frequency sound-coupled tasks, similar to control mice. Ex vivo cochlear Ca2+ imaging experiments demonstrated that without Prestin, the OHCs still exhibit ultrahigh-frequency transduction, which in contrast, can be abolished by a universal cation channel blocker, Gadolinium. In vivo salicylate treatment disrupts hearing at frequencies <40 kHz but not ultrahigh-frequency hearing. By pharmacogenetic manipulation, we showed that specific ablation of the OHCs largely abolished hearing at frequencies >40 kHz. These findings demonstrate that cochlear OHCs are the target cells that support ultrahigh-frequency transduction, which does not require Prestin.

Brain-wide projection reconstruction of single functionally defined neurons.

Reconstructing axonal projections of single neurons at the whole-brain level is currently a converging goal of the neuroscience community that is fundamental for understanding the logic of information flow in the brain. Thousands of single neurons from different brain regions have recently been morphologically reconstructed, but the corresponding physiological functional features of these reconstructed neurons are unclear. By combining two-photon Ca2+ imaging with targeted single-cell plasmid electroporation, we reconstruct the brain-wide morphologies of single neurons that are defined by a sound-evoked response map in the auditory cortices (AUDs) of awake mice. Long-range interhemispheric projections can be reliably labelled via co-injection with an adeno-associated virus, which enables enhanced expression of indicator protein in the targeted neurons. Here we show that this method avoids the randomness and ambiguity of conventional methods of neuronal morphological reconstruction, offering an avenue for developing a precise one-to-one map of neuronal projection patterns and physiological functional features.

Fibroblast Activation Protein-α as a Target in the Bench-to-Bedside Diagnosis and Treatment of Tumors: A Narrative Review.

Fibroblast activation protein-α (FAP) is a type II integral serine protease that is specifically expressed by activated fibroblasts. Cancer-associated fibroblasts (CAFs) in the tumor stroma have an abundant and stable expression of FAP, which plays an important role in promoting tumor growth, invasion, metastasis, and immunosuppression. For example, in females with a high incidence of breast cancer, CAFs account for 50-70% of the cells in the tumor's microenvironment. CAF overexpression of FAP promotes tumor development and metastasis by influencing extracellular matrix remodeling, intracellular signaling, angiogenesis, epithelial-to-mesenchymal transition, and immunosuppression. This review discusses the basic biological characteristics of FAP and its applications in the diagnosis and treatment of various cancers. We review the emerging basic and clinical research data regarding the use of nanomaterials that target FAP.

PIEZO2 mediates ultrasonic hearing via cochlear outer hair cells in mice.

Ultrasonic hearing and vocalization are the physiological mechanisms controlling echolocation used in hunting and navigation by microbats and bottleneck dolphins and for social communication by mice and rats. The molecular and cellular basis for ultrasonic hearing is as yet unknown. Here, we show that knockout of the mechanosensitive ion channel PIEZO2 in cochlea disrupts ultrasonic- but not low-frequency hearing in mice, as shown by audiometry and acoustically associative freezing behavior. Deletion of Piezo2 in outer hair cells (OHCs) specifically abolishes associative learning in mice during hearing exposure at ultrasonic frequencies. Ex vivo cochlear Ca2+ imaging has revealed that ultrasonic transduction requires both PIEZO2 and the hair-cell mechanotransduction channel. The present study demonstrates that OHCs serve as effector cells, combining with PIEZO2 as an essential molecule for ultrasonic hearing in mice.

In situ photoconductivity measurements of imidazole in optical fiber break-junctions.

We developed a method based on the mechanically controllable break junction technique to investigate the electron transport properties of single molecular junctions upon fiber waveguided light. In our strategy, a metal-coated tapered optical fiber is fixed on a flexible substrate, and this tapered fiber serves as both the optical waveguide and metal electrodes after it breaks. For an imidazole bridged single-molecule junction, two probable conductance values below 1G0 are observed. The higher value shows an approximately 40% enhancement under illumination, while the lower one does not show distinguishable difference under illumination. Theoretical calculations reveal these two conductance values resulting from the imidazole monomer junction and the imidazole dimer junction linked via a hydrogen bond, respectively. In imidazole monomer junctions, the absorption of a single photon strongly shifts the transmission function resulting in optical-induced conductance enhancement. In contrast, the transmission function of imidazole dimer junctions remains at the same level in the bias window despite the light illumination. This work provides a robust experimental framework for studying the underlying mechanisms of photoconductivity in single-molecule junctions and offers tools for tuning the optoelectronic performance of single-molecule devices in situ.

Single-neuron representation of learned complex sounds in the auditory cortex.

The sensory responses of cortical neuronal populations following training have been extensively studied. However, the spike firing properties of individual cortical neurons following training remain unknown. Here, we have combined two-photon Ca2+ imaging and single-cell electrophysiology in awake behaving mice following auditory associative training. We find a sparse set (~5%) of layer 2/3 neurons in the primary auditory cortex, each of which reliably exhibits high-rate prolonged burst firing responses to the trained sound. Such bursts are largely absent in the auditory cortex of untrained mice. Strikingly, in mice trained with different multitone chords, we discover distinct subsets of neurons that exhibit bursting responses specifically to a chord but neither to any constituent tone nor to the other chord. Thus, our results demonstrate an integrated representation of learned complex sounds in a small subset of cortical neurons.

Simultaneous Desulfurization and Denitrification Using La-Ce-V-Cu-ZSM-5 Catalysts in an Electrostatic Precipitator.

Different catalysts were loaded onto the collecting plate of an electrostatic precipitator to achieve the simultaneous removal of multiple pollutants from coal-fired gas. The synergistic desulfurization and denitrification effect of the catalyst and the effect of corona discharge on the activity of the catalyst were studied. The La(6%)-Ce(8%)-V(7%)-Cu(8%)-ZSM-5 catalyst prepared by successive impregnation methods had the optimum simultaneous desulfurization and denitrification efficiency at a roasting temperature of 600 °C. The desulfurization and denitrification rates reached 97.09 and 83.30%, respectively. BET and SEM characterization results showed that the loading of active components and additives improved the pore structure of the molecular sieve, which contributed to the high stability of the catalyst's internal structure and large surface area, as well as better desulfurization and denitrification efficiency. Corona discharge can significantly improve the catalytic effect.

Author Correction: Dual-energy CT perfusion imaging for differentiating WHO subtypes of thymic epithelial tumors.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Dual-energy CT perfusion imaging for differentiating WHO subtypes of thymic epithelial tumors.

To evaluate the role of conventional contrast-enhanced CT (CECT) imaging and dual-energy spectral CT (DECT) perfusion imaging in differentiating the WHO histological subtypes of thymic epithelial tumours (TETs). Eighty-eight patients with TETs who underwent DECT perfusion scans (n = 51) and conventional CT enhancement scans (n = 37) using a GE Discovery CT750 HD scanner were enrolled in this study. The mean maximal contrast-enhanced range (mean CEmax) and the perfusion and spectral parameters of the lesions were analysed. Among the six WHO subtypes (Type A, AB, B1, B2, and B3 thymoma and thymic carcinoma), the mean CEmax values and most of the perfusion and spectral parameter values of Type A and Type AB were significantly higher than those of the other subtypes (all P < 0.05), and there was no difference among Type B1, B2 and B3 (all P > 0.05). The mean CEmax value was not different between Type B (including Type B1, B2, and B3) and thymic carcinoma (P = 1.000). The PS, IC, NIC and λHU values in the optimal venous phase of thymic carcinoma were higher than those of Type B (all P < 0.05). The parameters of conventional CECT imaging and DECT perfusion imaging can help identify the subtype of TETs, especially those of DECT perfusion imaging in type B thymomas and thymic carcinomas.

Atomic switches of metallic point contacts by plasmonic heating.

Electronic switches with nanoscale dimensions satisfy an urgent demand for further device miniaturization. A recent heavily investigated approach for nanoswitches is the use of molecular junctions that employ photochromic molecules that toggle between two distinct isoforms. In contrast to the reports on this approach, we demonstrate that the conductance switch behavior can be realized with only a bare metallic contact without any molecules under light illumination. We demonstrate that the conductance of bare metallic quantum contacts can be reversibly switched over eight orders of magnitude, which substantially exceeds the performance of molecular switches. After the switch process, the gap size between two electrodes can be precisely adjusted with subangstrom accuracy by controlling the light intensity or polarization. Supported by simulations, we reveal a more general and straightforward mechanism for nanoswitching behavior, i.e., atomic switches can be realized by the expansion of nanoelectrodes due to plasmonic heating.

Neuregulin-1 Accelerates Functional Motor Recovery by Improving Motoneuron Survival After Brachial Plexus Root Avulsion in Mice.

Brachial plexus root avulsion (BPRA) results in the complete loss of motor function in the upper limb, mainly due to the death of spinal motoneurons (MNs). The survival of spinal MNs is the key to the recovery of motor function. Neuregulin-1 (Nrg1) plays fundamental roles in nervous system development and nerve repair. However, its functional role in BPRA remains unclear. On the basis of our findings that Nrg1 is down-regulated in the ventral horn in a mouse model of BPRA, Nrg1 may be associated with BPRA. Here, we investigated whether recombinant Nrg1ß (rNrg1ß) can enhance the survival of spinal MNs and improve functional recovery in mice following BPRA. In vitro studies on primary cultured mouse MNs showed that rNrg1ß increased the survival rate in a dose-dependent manner, reaching a peak at 5 nM, which increased the survival rate and enhanced the pERK levels in MNs under H2O2-induced oxidative stress. In vivo studies revealed that rNrg1ß improved the functional recovery of elbow flexion, promoted the survival of MNs, enhanced the re-innervation of biceps brachii, and decreased the muscle atrophy. These results suggest that Nrg1 may provide a potential therapeutic strategy for root avulsion.