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At least 1,783 records · Page 99

Muscone suppresses inflammatory responses and neuronal damage in a rat model of cervical spondylotic myelopathy by regulating Drp1‐dependent mitochondrial fission

Abstract Cervical spondylotic myelopathy (CSM) is a common cause of disability with few treatments. Aberrant mitochondrial dynamics play a crucial role in the pathogenesis of various neurodegenerative diseases. Thus, regulation of mitochondrial dynamics may offer therapeutic benefit for the treatment of CSM. Muscone, the active ingredient of an odoriferous animal product, exhibits anti‐inflammatory and neuroprotective effects for which the underlying mechanisms remain obscure. We hypothesized that muscone might ameliorate inflammatory responses and neuronal damage by regulating mitochondrial dynamics. To this end, the effects of muscone on a rat model of chronic cervical cord compression, as well as activated BV2 cells and injured neurons, were assessed. The results showed that muscone intervention improved motor function compared with vehicle‐treated rats. Indeed, muscone attenuated pro‐inflammatory cytokine expression, neuronal‐apoptosis indicators in the lesion area, and activation of the nod‐like receptor family pyrin domain‐containing 3 inflammasome, nuclear transcription factor‐κB, and dynamin‐related protein 1 in Iba1‐ and βIII‐tubulin‐labeled cells. Compared with vehicle‐treated rats, compression sites of muscone‐treated animals exhibited elongated mitochondrial morphologies in individual cell types and reduced reactive oxygen species. In vitro results indicated that muscone suppressed microglial activation and neuronal damage by regulating related‐inflammatory or apoptotic molecules. Moreover, muscone inhibited dynamin‐related protein 1 activation in activated BV2 cells and injured neurons, whereby it rescued mitochondrial fragmentation and reactive oxygen species production, which regulate a wide range of inflammatory and apoptotic molecules. Our findings reveal that muscone attenuates neuroinflammation and neuronal damage in rats with chronic cervical cord compression by regulating mitochondrial fission events, suggesting its promise for CSM therapy. image

Zhou, Long‐yun↗

Identification of new marker genes from plant single‐cell RNA‐seq data using interpretable machine learning methods

Summary An essential step in the analysis of single‐cell RNA sequencing data is to classify cells into specific cell types using marker genes. In this study, we have developed a machine learning pipeline called single‐cell predictive marker (SPmarker) to identify novel cell‐type marker genes in the Arabidopsis root. Unlike traditional approaches, our method uses interpretable machine learning models to select marker genes. We have demonstrated that our method can: assign cell types based on cells that were labelled using published methods; project cell types identified by trajectory analysis from one data set to other data sets; and assign cell types based on internal GFP markers. Using SPmarker, we have identified hundreds of new marker genes that were not identified before. As compared to known marker genes, the new marker genes have more orthologous genes identifiable in the corresponding rice single‐cell clusters. The new root hair marker genes also include 172 genes with orthologs expressed in root hair cells in five non‐ Arabidopsis species, which expands the number of marker genes for this cell type by 35–154%. Our results represent a new approach to identifying cell‐type marker genes from scRNA‐seq data and pave the way for cross‐species mapping of scRNA‐seq data in plants.

54 ENVIRONMENTAL SCIENCES↗

The soil pore structure encountered by roots affects plant‐derived carbon inputs and fate

Summary Plant roots are the main supplier of carbon (C) to the soil, the largest terrestrial C reservoir. Soil pore structure drives root growth, yet how it affects belowground C inputs remains a critical knowledge gap. By combining X‐ray computed tomography with 14 C plant labelling, we identified root–soil contact as a previously unrecognised influence on belowground plant C allocations and on the fate of plant‐derived C in the soil. Greater contact with the surrounding soil, when the growing root encounters a pore structure dominated by small (< 40 μm Ø) pores, results in strong rhizodeposition but in areas of high microbial activity. The root system of Rudbeckia hirta revealed high plasticity and thus maintained high root–soil contact. This led to greater C inputs across a wide range of soil pore structures. The root–soil contact Panicum virgatum , a promising bioenergy feedstock crop, was sensitive to the encountered structure. Pore structure built by a polyculture, for example, restored prairie, can be particularly effective in promoting lateral root growth and thus root–soil contact and associated C benefits. The findings suggest that the interaction of pore structure with roots is an important, previously unrecognised, stimulus of soil C gains.

Plant Sciences↗

Arbuscular mycorrhiza convey significant plant carbon to a diverse hyphosphere microbial food web and mineral‐associated organic matter

Summary Arbuscular mycorrhizal fungi (AMF) transport substantial plant carbon (C) that serves as a substrate for soil organisms, a precursor of soil organic matter (SOM), and a driver of soil microbial dynamics. Using two‐chamber microcosms where an air gap isolated AMF from roots, we 13 CO 2 ‐labeled Avena barbata for 6 wk and measured the C Rhizophagus intraradices transferred to SOM and hyphosphere microorganisms. NanoSIMS imaging revealed hyphae and roots had similar 13 C enrichment. SOM density fractionation, 13 C NMR, and IRMS showed AMF transferred 0.77 mg C g −1 of soil (increasing total C by 2% relative to non‐mycorrhizal controls); 33% was found in occluded or mineral‐associated pools. In the AMF hyphosphere, there was no overall change in community diversity but 36 bacterial ASVs significantly changed in relative abundance. With stable isotope probing (SIP)‐enabled shotgun sequencing, we found taxa from the Solibacterales, Sphingobacteriales, Myxococcales, and Nitrososphaerales (ammonium oxidizing archaea) were highly enriched in AMF‐imported 13 C (> 20 atom%). Mapping sequences from 13 C‐SIP metagenomes to total ASVs showed at least 92 bacteria and archaea were significantly 13 C‐enriched. Our results illustrate the quantitative and ecological impact of hyphal C transport on the formation of potentially protective SOM pools and microbial roles in the AMF hyphosphere soil food web.

13C SIP↗

Cell wall ester modifications and volatile emission signatures of plant response to abiotic stress

Abstract Growth suppression and defence signalling are simultaneous strategies that plants invoke to respond to abiotic stress. Here, we show that the drought stress response of poplar trees ( Populus trichocarpa ) is initiated by a suppression in cell wall derived methanol (MeOH) emissions and activation of acetic acid (AA) fermentation defences. Temperature sensitive emissions dominated by MeOH (AA/MeOH <30%) were observed from physiologically active leaves, branches, detached stems, leaf cell wall isolations and whole ecosystems. In contrast, drought treatment resulted in a suppression of MeOH emissions and strong enhancement in AA emissions together with volatiles acetaldehyde, ethanol, and acetone. These drought‐induced changes coincided with a reduction in stomatal conductance, photosynthesis, transpiration, and leaf water potential. The strong enhancement in AA/MeOH emission ratios during drought (400%–3500%) was associated with an increase in acetate content of whole leaf cell walls, which became significantly 13 C 2 ‐labelled following the delivery of 13 C 2 ‐acetate via the transpiration stream. The results are consistent with both enzymatic and nonenzymatic MeOH and AA production at high temperature in hydrated tissues associated with accelerated primary cell wall growth processes, which are downregulated during drought. While the metabolic source(s) require further investigation, the observations are consistent with drought‐induced activation of aerobic fermentation driving high rates of foliar AA emissions and enhancements in leaf cell wall O ‐acetylation. We suggest that atmospheric AA/MeOH emission ratios could be useful as a highly sensitive signal in studies investigating environmental and biological factors influencing growth‐defence trade‐offs in plants and ecosystems.

59 BASIC BIOLOGICAL SCIENCES↗

Virocell Necromass Provides Limited Plant Nitrogen and Elicits Rhizosphere Metabolites That Affect Phage Dynamics

Bacteriophages impact soil bacteria through lysis, altering the availability of organic carbon and plant nutrients. However, the magnitude of nutrient uptake by plants from lysed bacteria remains unknown, partly because this process is challenging to investigate in the field. In this study, we extend ecosystem fabrication (EcoFAB 2.0) approaches to study plant-bacteria-phage interactions by comparing the impact of virocell (phage-lysed) and uninfected 15 N-labelled bacterial necromass on plant nitrogen acquisition and rhizosphere exometabolites composition. We show that grass Brachypodium distachyon derives some nitrogen from amino acids in uninfected Pseudomonas putida necromass lysed by sonication but not from virocell necromass. Additionally, the bacterial necromass elicits the formation of rhizosphere exometabolites, some of which (guanosine), alongside tested aromatic acids ( p -coumaric and benzoic acid), show bacterium-specific effects on bacteriophage-induced lysis when tested in vitro. The study highlights the dynamic feedback between virocell necromass and plants and suggests that root exudate metabolites can impact bacteriophage infection dynamics.

Brachypodium↗

Root‐Pore Interactions, the Underestimated Driver for Rhizosphere Structure and Rhizosheath Development

Physical characteristics of rhizosphere and rhizosheath, that is, root-adhering soil, are crucial for plant performance. Yet, the drivers of the rhizosphere's structural properties and their relationships with rhizosheath development remain unclear. We used X-ray computed micro-tomography (i) to explore two drivers of rhizosphere porosity: root-induced changes vs. preferential root growth into soil with certain pore characteristics and (ii) to estimate their contributions to rhizosphere macroporosity gradients and rhizosheath formation. Rhizosheath development was assessed in relation to rhizosphere macroporosity and rhizodeposition after ¹⁴C labeling. Our results confirmed that both root-induced changes and growth preferences shape rhizosphere structure, with their relative significance depending on the inherent macropore availability. In intact soils, growth preferences were the dominant factor, while in sieved soils the root-induced changes became equally important. Rhizosheath formation was associated with roots compacting their surrounding and releasing carbon. However, no correlation was found between rhizosheath formation and the actual rhizosphere, that is, the volume of soil adjacent to the roots. The study offers new process-level understanding of rhizosphere porosity gradients, while emphasizing caution in interpreting root growth data from sieved soil studies. Similarly, traditional destructively sampled rhizosheath may not fully capture the true characteristics of the actual rhizosphere, underscoring importance of intact-soil analyses.

macroporosity gradients↗

A chloroplast diacylglycerol lipase modulates glycerolipid pathway balance in Arabidopsis

SUMMARY Two parallel pathways compartmentalized in the chloroplast and the endoplasmic reticulum contribute to thylakoid lipid synthesis in plants, but how these two pathways are coordinated during thylakoid biogenesis and remodeling remains unknown. We report here the molecular characterization of a homologous ADIPOSE TRIGLYCERIDE LIPASE‐LIKE gene, previously referred to as ATGLL . The ATGLL gene is ubiquitously expressed throughout development and rapidly upregulated in response to a wide range of environmental cues. We show that ATGLL is a chloroplast non‐regioselective lipase with a hydrolytic activity preferentially towards 16:0 of diacylglycerol (DAG). Comprehensive lipid profiling and radiotracer labeling studies revealed a negative correlation of ATGLL expression and the relative contribution of the chloroplast lipid pathway to thylakoid lipid biosynthesis. Additionally, we show that genetic manipulation of ATGLL expression resulted in changes in triacylglycerol levels in leaves. We propose that ATGLL, through affecting the level of prokaryotic DAG in the chloroplast, plays important roles in balancing the two glycerolipid pathways and in maintaining lipid homeostasis in plants.

59 BASIC BIOLOGICAL SCIENCES↗

Deep Learning for Intelligent Bubble Size Detection in the Spallation Neutron Source Visual Target

The Spallation Neutron Source (SNS) at Oak Ridge National Laboratory (ORNL) will undergo proton power upgrade (PPU), increasing the proton beam power from 1.4 MW to 2.8 MW. From 2.8 MW, 2.0 MW will go to the current First Target Station and the rest will go to the future Second Target Station (STS). The First Target Station uses a liquid mercury target that is contained in a 316L stainless steel vessel. The proton beam is pulsed at 60 Hz, with a pulse of about 0.7μs. When the proton beam hits the target, the intense energy deposition leads to a rapid rise in temperature in the mercury. This temperature rise creates pressure waves that propagate through the mercury and cause cavitation erosion. The power upgrade will cause stronger pressure waves that will further increase damage because of cavitation. Injecting small helium bubbles in the mercury has been an efficient method of mitigating the pressure wave at 1.4 MW. However, at higher power, additional mitigation is necessary. Therefore, the 2 MW target vessel will be equipped with swirl bubblers and an additional gas injection port near the nose to inject more gas in the target. To develop a gas injection strategy and design, flow visualization in water with a transparent prototypical target (“visual target”) was performed. Bubble sizes and their spatial distribution in the flow loop are crucial to understanding the effectiveness of the bubbles in mitigating pressure waves. Bubbles were generated in the visual target under varied conditions of input pressures with helium and air. Images were captured using a high-speed camera at varied frame rates at different positions away from the swirl bubbler and different depths in the flow loop under varying lighting conditions. Initially, methods such as circular Hough transforms were applied after a series of images processing to obtain a general distribution of bubble sizes. Bubbles smaller than 500 μm are preferred to effectively mitigate the effect of pressure waves, which demands an accurate bubble detection and sizing system. Intelligent detection and identification of bubble sizes alleviate misdetection and improves accuracies. Employing neural networks, intelligent detection of bubble sizes and their distribution was developed and provides a robust alternative to traditional techniques. Human intervention was employed to label in-focus and out-of-focus bubbles in the set of training images. An object detection network using a pretrained convolutional neural network was created that extracted the features from the training images. Data augmentation was used to improve network accuracy through a random transformation of the original data.

Rasheed, Fayaz↗

Quantifying Trapped Powder in Electron Beam Powder Bed Fusion

Abstract Electron beam powder bed fusion (PBF-EB) shows great potential for manufacturing complex parts including those with internal cavities for heat exchanger, manifold systems, or energy absorption purposes. PBF-EB allows for the manufacture of channel geometries without the need for support structures. Due to the nature of the powder spreading process, powder feedstock is often trapped in intentionally manufactured cavities. This trapped powder can often be difficult to remove and can disturb the intended flow of fluid through the cavity or damage downstream components in its use case. These trapped powder particles present a risk of contamination and component failure if not completely evacuated. Ti6Al4V is a choice material for aerospace applications due to its high strength to weight ratio and its composition as a nonferrous metal; however, in weight sensitive applications excess entrapped powders or powders loosely attached to the surface could cause undesirable weight increases. The inherent spreading process of PBF-EB is different than laser powder bed fusion (PBF-LB) in its operational temperature, sintering. In addition, PBF-EB is less commonly studied in literature compared to its PBF-LB counterpart, and as a result the complexity of the semi-sintered powder and its spreading behavior are not well understood. Prior work has investigated the difficulty in removing trapped powder from PBF-EB, but these studies do not address how to quantify the amount of trapped powder in the cavity. Thus, an accurate method to measure the amount of trapped powder in the cavity must be investigated. In this work, Ti6Al4V coupons were manufactured with horizontal and vertical cavities of three different sizes. Archimedes testing allows for the determination of density differences caused by porosity and trapped powders by measuring mass and volumetric dispersion. Computed tomography (CT) is well suited for segmenting the internal structure and features of a part and has been studied for applications including voids, porosity, and dross. Thus, CT was explored as a method for evaluating trapped powder content in this work. The volumetric representation of the segmentation of the reconstructed CT volume can vary greatly depending on the input filter and thresholding methods. In this study, four different types of segmentation approaches were evaluated to determine the best approach for segmenting the volume as compared to an operator labeled ground truth. The percentage density results from the Archimedes testing were compared to the volumetric percent density from the computed tomography approach. Differences in packing density between two different internal channel features were investigated. Overall, this work sought to validate the use of computed tomography for the detection of trapped powders and present a framework for volumetric segmentation.

Johnstone, Brian↗

Adaptive noise canceling for transient absorption microscopy

Significance: Ultrafast fiber lasers are an attractive alternative to bulk lasers for nonlinear optical microscopy for their compactness and low cost. The high relative intensity noise (RIN) of these lasers poses a challenge for pump-probe measurements such as transient absorption and stimulated Raman scattering, along with modalities that provide label-free contrast from the vibrational and electronic structure of molecules. Aim: Digital adaptive filtering was applied to determine the applicability for canceling laser RIN in a transient absorption microscope with an ultrafast fiber laser source. Approach: Digitized signals from the transmitted probe and reference photodetectors were fed to an adaptive filter in MATLAB, running in a noise canceling configuration. This result was then fed to a software lock-in algorithm to demodulate the pump-probe signal. Images were built up one line scan at a time with a 3.5-kHz resonant scanner, with 100× averaging. The imaging target was Bi4Ge3O12, which exhibits nondegenerate two-photon absorption at the pump/probe wavelengths used (530-nm pump and 490-nm probe). Results: Without adaptive noise cancellation, the lock-in output primarily passes the laser RIN within its detection bandwidth, resulting in images that closely follow the linear transmissivity and lack sensitivity to pump-probe time delay. With adaptive noise cancellation in front of the lock-in, the RIN rejection is enough to restore the z-sectioning and sensitivity to pump-probe delay, as expected for transient absorption. Results were limited primarily by noise from the photodetector and analog-to-digital converter. Conclusions: Digital adaptive noise cancellation, even when limited by electronics noise, can recover pump-probe signals by removal of laser RIN, under conditions where averaging alone fails.

42 ENGINEERING↗

Survey of Image Denoising Methods for Medical Image Classification

Medical imaging devices, such as X-ray machines, inherently produce images that suffer from visual noise. Our objectives were to (i.) determine the effect of image denoising on a medical image classification task, and (ii.) determine if there exists a correlation between image denoising performance and medical image classification performance. We performed the medical image classification task on chest X-rays using the DenseNet-121 convolutional neural network (CNN) and used the peak signal-to-noise ratio (PSNR) and structural similarity (SSIM) metrics as the image denoising performance measures. We first found that different denoising methods can make a statistically significant difference in classification performance for select labels. We also found that denoising methods affect fine-tuned models more than randomly-initialized models and that fine-tuned models have significantly higher and more uniform performance than randomly-initialized models. Lastly, we found that there is no significant correlation between PSNR and SSIM values and classification performance for our task.

Michael, Peter↗

Training data selection for event classification in a highly variable environment

A problem of interest for nuclear nonproliferation is monitoring activities at nuclear facilities, where proliferation events may only take place a few times and often under variable conditions. Machine learning has revolutionized data analytics by enabling the use of measurable signatures to generate predictive models of facility operations. However, traditional methods for training these models require large, reliable data sets with labeled observations, a challenge for nonproliferation. Highly variable conditions further complicate this as events from training data may have occurred in conditions quite different from the event of interest. Our hypothesis is that when events occur in a highly variable environment, careful training data selection for each test event could outperform the standard approach of using all available training data. We developed a method to optimize training data selection for the given test event and applied it to predicting the power level of the High Flux Isotope Reactor (HFIR) at Oak Ridge National Laboratory. In this study, the reactor startup exhibits variability between occurrences due to natural variability in environmental conditions and operational procedures. Using a combination of analysis techniques, a similitude assessment was performed on data collected from HFIR to isolate clusters that were optimal for training a predictive model. Concepts such as dynamic time warping and Jaccard similarity were used in conjunction with clustering analysis. In order to validate this approach, the model was trained on every combination of unique training events and the predictive performance was compared to the performance using a subset of the training data selected by isolated clusters found through the similitude assessment.

Iyer, A↗

Nanoscale chromatin imaging and analysis platform bridges 4D chromatin organization with molecular function

Extending across multiple length scales, dynamic chromatin structure is linked to transcription through the regulation of genome organization. However, no individual technique can fully elucidate this structure and its relation to molecular function at all length and time scales at both a single-cell level and a population level. Here, we present a multitechnique nanoscale chromatin imaging and analysis (nano-ChIA) platform that consolidates electron tomography of the primary chromatin fiber, optical super-resolution imaging of transcription processes, and label-free nano-sensing of chromatin packing and its dynamics in live cells. Using nano-ChIA, we observed that chromatin is localized into spatially separable packing domains, with an average diameter of around 200 nanometers, sub-megabase genomic size, and an internal fractal structure. The chromatin packing behavior of these domains exhibits a complex bidirectional relationship with active gene transcription. Furthermore, we found that properties of PDs are correlated among progenitor and progeny cells across cell division.

59 BASIC BIOLOGICAL SCIENCES↗

Deconvolution of metal apportionment in bulk metal-organic frameworks

We report a general route to decipher the apportionment of metal ions in bulk metal-organic frameworks (MOFs) by solid-state nuclear magnetic resonance spectroscopy. We demonstrate this route in Mg 1-x Ni x -MOF-74, where we uncover all eight possible atomic-scale Mg/Ni arrangements through identification and quantification of the distinct chemical environments of 13 C-labeled carboxylates as a function of the Ni content. Here, we use magnetic susceptibility, bond pathway, and density functional theory calculations to identify local metal bonding configurations. The results refute the notion of random apportionment from solution synthesis; rather, we reveal that only two of eight Mg/Ni arrangements are preferred in the Ni-incorporated MOFs. These preferred structural arrangements manifest themselves in macroscopic adsorption phenomena as illustrated by CO/CO 2 breakthrough curves. We envision that this nondestructive methodology can be further applied to analyze bulk assembly of other mixed-metal MOFs, greatly extending the knowledge on structure-property relationships of MOFs and their derived materials.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Molecular insights into the evolution of woody plant decay in the gut of termites

Plant cell walls represent the most abundant pool of organic carbon in terrestrial ecosystems but are highly recalcitrant to utilization by microbes and herbivores owing to the physical and chemical barrier provided by lignin biopolymers. Termites are a paradigmatic example of an organism’s having evolved the ability to substantially degrade lignified woody plants, yet atomic-scale characterization of lignin depolymerization by termites remains elusive. We report that the phylogenetically derived termite Nasutitermes sp. efficiently degrades lignin via substantial depletion of major interunit linkages and methoxyls by combining isotope-labeled feeding experiments and solution-state and solid-state nuclear magnetic resonance spectroscopy. Exploring the evolutionary origin of lignin depolymerization in termites, we reveal that the early-diverging woodroach Cryptocercus darwini has limited capability in degrading lignocellulose, leaving most polysaccharides intact. Conversely, the phylogenetically basal lineages of “lower” termites are able to disrupt the lignin-polysaccharide inter- and intramolecular bonding while leaving lignin largely intact. These findings advance knowledge on the elusive but efficient delignification in natural systems with implications for next-generation ligninolytic agents.

59 BASIC BIOLOGICAL SCIENCES↗

Imaging beyond the surface region: Probing hidden materials via atomic force microscopy

Probing material properties at surfaces down to the single-particle scale of atoms and molecules has been achieved, but high-resolution subsurface imaging remains a nanometrology challenge due to electromagnetic and acoustic dispersion and diffraction. The atomically sharp probe used in scanning probe microscopy (SPM) has broken these limits at surfaces. Subsurface imaging is possible under certain physical, chemical, electrical, and thermal gradients present in the material. Of all the SPM techniques, atomic force microscopy has entertained unique opportunities for nondestructive and label-free measurements. Here, we explore the physics of the subsurface imaging problem and the emerging solutions that offer exceptional potential for visualization. We discuss materials science, electronics, biology, polymer and composite sciences, and emerging quantum sensing and quantum bio-imaging applications. The perspectives and prospects of subsurface techniques are presented to stimulate further work toward enabling noninvasive high spatial and spectral resolution investigation of materials including meta- and quantum materials.

74 ATOMIC AND MOLECULAR PHYSICS↗

Persistent urban heat

Urban surface and near-surface air temperatures are known to be often higher than their rural counterparts, a phenomenon now labeled as the urban heat island effect. However, whether the elevated urban temperatures are more persistent than rural temperatures at timescales commensurate to heat waves has not been addressed despite its importance for human health. Combining numerical simulations by a global climate model with a surface energy balance theory, it is demonstrated here that urban surface and near-surface air temperatures are significantly more persistent than their rural counterparts in cities dominated by impervious materials with large thermal inertia. Further use of these materials will result in even stronger urban temperature persistence, especially for tropical cities. The present findings help pinpoint mitigation strategies that can simultaneously ameliorate the larger magnitude and stronger persistence of urban temperatures.

54 ENVIRONMENTAL SCIENCES↗