Quantitative Electron Microscopy Research Articles

Complete miscibility of immiscible elements at the nanometre scale.

Understanding the mixing behaviour of elements in a multielement material is important to control its structure and property. When the size of a multielement material is decreased to the nanoscale, the miscibility of elements in the nanomaterial often changes from its bulk counterpart. However, there is a lack of comprehensive and quantitative experimental insight into this process. Here we explored how the miscibility of Au and Rh evolves in nanoparticles of sizes varying from 4 to 1 nm and composition changing from 15% Au to 85% Au. We found that the two immiscible elements exhibit a phase-separation-to-alloy transition in nanoparticles with decreased size and become completely miscible in sub-2 nm particles across the entire compositional range. Quantitative electron microscopy analysis and theoretical calculations were used to show that the observed immiscibility-to-miscibility transition is dictated by particle size, composition and possible surface adsorbates present under the synthesis conditions.

Effect of gypsum content in cement on its adaptability to alkali-free accelerator at different temperatures

The setting times and early strength of shotcrete are greatly affected by the gypsum content of the cement used. In this work, the setting times, compressive strength and microscopic analysis (X-ray diffraction quantitative analysis, thermogravimetric analysis and scanning electron microscopy) were used to determine the effects of changes in gypsum content on the hydration and mechanical properties of cement mixed with an accelerator at different temperatures. The results showed that an increase in the gypsum content of the cement promoted the formation of ettringite (AFt) and decreased the setting times of the cement pastes at 0°C, 20°C and 40°C. Mortar compressive strength tests showed that, after curing at 0°C, 20°C and 40°C, the highest compressive strength was exhibited with a final molar aluminate-to-sulfate ratio (C3A/SO3; henceforth called the MASR) of 0.84, 0.84 and 1.18, respectively. The mechanical properties of the hardened cement pastes were adversely affected by MASRs that were too high or too low. When the MASR was too high, the conversion of AFt to monosulfate (AFm) was facilitated and C3S hydration was slowed. Conversely, when the MASR was too low, this enabled large amounts of AFt to occupy the space where calcium silicate hydrate (C-S-H) gels were formed, resulting in decalcification of the C-S-H gels.

Microstructure-Based Creep Life Assessment of 1CrMoV Turbine Rotor Steels After Long-Term Service

This study evaluates the use of quantitative analytical electron microscopy for microstructure-based creep life assessment of service-exposed 1CrMoV steel turbine rotors. Changes in the microstructure (bainitic laths, carbide phases) were related to creep life exhaustion estimations done using conventional methods based on cavities and hardness. The volume-weighted average size and surface density of bainitic laths correlate with cavity-based estimated creep life exhaustion. However, the heterogeneity of grain structure limits the use of bainitic lath parameters for assessing creep life based on microstructure. The phase proportions of M3C, M2C, and M7C3 carbides, as determined by TKD-EDS mapping, show a strong correlation (R2: 0.64, 0.61, and 0.86) with creep life exhaustion estimations and could potentially be used as an additional indicator of the material state of the critical failure region in 1CrMoV turbine rotors.

A study of fluid overpressure microstructures from the creeping segment of the San Andreas fault

Evidence of episodic fluid overpressure events noted in samples from the San Andreas Fault Observatory at Depth (SAFOD) have remained largely uncorrelated in terms of their collective significance for seismic history of the fault zone. The compositional and microstructural correlations sought in this study could shed light on questions about potential for major seismic events in the creeping segment of the SAF in central California. We used quantitative energy dispersive spectroscopy (EDS), Cathodoluminescence (CL) and Scanning Electron Microscope (SEM) imaging, and electron backscatter diffraction (EBSD) analysis to acquire geochemical and microstructural data from a suite of twenty SAFOD core samples including the damage zone and the active core of the fault. The results indicate intermittent coseismic fluid overpressure events that overprint the background aseismic creep across the fault. Analysis of trace elements and deformation in the coseismic calcite vein generations and their associated hydrothermal mineral phases indicate progressive uplift and exhumation followed by an asymmetric incursion of meteoric water into the damage zone. The same analysis suggests that the actively creeping intervals act as permeability barriers. Our results are in overall agreement with recent studies of the SAF in central California that indicate large seismic events have occurred intermittent with aseismic creep in recent geological time or suggest future potential for such events.

BNIP3 in hypoxia-induced mitophagy: Novel insights and promising target for non-alcoholic fatty liver disease

BNIP3 localizes to the outer mitochondrial membrane, has been demonstrated to be extensively involved in abnormalities to mitochondrial metabolic function and dynamicsand in non-alcoholic fatty liver disease (NAFLD). However, its role in NAFLD under hypoxia remains unclear. This study aimed to investigate the expression and the role of BNIP3 in NAFLD under hypoxia, and explore its involvement in regulating NAFLD mitophagy, fatty acid β-oxidation both in vivo and in vitro. BNIP3-mediated mitophagy level was analyzed using real-time quantitative polymerase chain reaction, Western blotting, immunofluorescence and electron microscopy. The role of BNIP3 in fatty acid β-oxidation was evaluated using lipid droplet staining, triglyceride content determination, and cellular energy metabolism. The results showed that compared with the HFD-2200 m, the body weight, inflammatory liver injury, and lipid deposition were significantly reduced in the HFD-4500 m group (P < 0.05), but autophagy and mitophagy were increased, and the expression of the mitophagy receptor BNIP3 was increased (P < 0.05). Compared to the control group, BNIP3 knockdown in the hypoxia group resulted in decreased levels of CPT1, ATGL, and p-HSL in lipid-accumulating hepatocytes, lipid droplet accumulation and triglyceride content increased (P < 0.05). Moreover, the ability of lipid-accumulating hepatocytes to oxidize fatty acids was reduced by BNIP3 knockdown in the hypoxia group (P < 0.05). Therefore, it can be concluded that, in NAFLD mice under hypoxia, BNIP3-mediated mitophagy promotes fatty acid β-oxidation. This study elucidated the role of BNIP3 in promoting fatty acid β-oxidation in NAFLD under hypoxia, and suggests BNIP3 may serve as a novel potential therapeutic target for NAFLD.

Electrolyzed Saline Targets Biofilm Periodontal Pathogens In Vitro

Preventing the development and recurrence of periodontal diseases often includes antimicrobial mouthrinses to control the growth of the periodontal pathogens. Most antimicrobials are nonselective, targeting the symbiotic oral species as well as the dysbiosis-inducing ones. This affects the overall microbial composition and metabolic activity and consequently the host–microbe interactions, which can be detrimental (associated with inflammation) or beneficial (health-associated). Consequently, guiding the antimicrobial effect for modulating the microbial composition to a health-associated one should be considered. For such an approach, this study investigated electrolyzed saline as a novel rinse. Electrolyzed saline was prepared from sterile saline using a portable electrolysis device. Multispecies oral homeostatic and dysbiotic biofilms were grown on hydroxyapatite discs and rinsed daily with electrolyzed saline (EOS). Corresponding positive (NaOCl) and negative (phosphate-buffered saline) controls were included. After 3 rinses, biofilms were analyzed with viability quantitative polymerase chain reaction and scanning electron microscopy. Supernatants of rinsed biofilms were used for metabolic activity analysis (high-performance liquid chromatography) through measuring organic acid content. In addition, human oral keratinocytes (HOKs) were exposed to EOS to test biocompatibility (cytotoxicity and inflammation induction) and also to rinsed biofilms to assess their immunogenicity after rinsing. Rinsing the dysbiotic biofilms with EOS could reduce the counts of the pathobionts (>3 log10 Geq/mm2 reduction) and avert biofilm dysbiosis (≤1% pathobiont abundance), leading to the dominance of commensal species (≥99%), which altered both biofilm metabolism and interleukin 8 (IL-8) induction in HOKs. EOS had no harmful effects on homeostatic biofilms. The scanning electron micrographs confirmed the same. In addition, tested concentrations of EOS did not have any cytotoxic effects and did not induce IL-8 production in HOKs. EOS showed promising results for diverting dysbiosis in in vitro rinsed biofilms and controlling key periopathogens, with no toxic effects on commensal species or human cells. This novel rinsing should be considered for clinical applications.

Vertical transmission and prevalence of white spot syndrome virus (WSSV) in the wild spawning population of the Indian white shrimp, Penaeus indicus

White spot disease, caused by white spot syndrome virus (WSSV), has historically been the most devastating disease in shrimp aquaculture industry across the world. The mode of virus transmission is the most crucial stage in the dynamics and management of virus infection. This study explored the mechanism of vertical transmission of WSSV in Indian white shrimp, Penaeus indicus, potential native species for domestication and genetic improvement, using quantitative real time PCR (q RT PCR), light and electron microscopy, and in situ hybridization. Wild brooders of P. indicus (n = 2576) were sampled along the South east coast of India, during 2016 to 2021. Of these ∼ 58 % of the brooders were positive for WSSV, and almost 50 % of infected wild brooders were at the various stages of reproductive maturation. WSSV-PCR positive brooders (n = 200) were analysed for vertical WSSV transmission. The q RT PCR studies of reproductive tissues revealed that 61 % (n = 13) of spermatophore, 54 % (n = 28) of immature ovaries and 48 % (n = 27) of ripe ovaries were infected with WSSV. The lowest level of infection was recorded in females with ripe ovaries (6.84 × 101 ± 9.79 × 100 ng genomic DNA) followed by fertilized eggs (1.59 × 102 ± 3.69 × 101 ng genomic DNA), and larvae (nauplius and zoea). The histology of gravid females with high WSSV copies showed pyknotic and karyorrhectic germinal vesicle with degenerated cortical rods. Conversely, the gravid females with low WSSV copies showed fully developed ovary without characteristic signs of WSSV infection. Transmission electron microscopic studies clearly established the presence of WSSV particles in both ovaries and spermatophores. When subjected to in situ hybridization, WSSV-specific signals were observed in connective tissues of spermatophore, although gravid ovary and fertilized eggs were failed to produce WSSV specific signals. The present study provides the first molecular and histological evidence for trans-ovarian vertical transmission of WSSV. Development of disease-free base population being the cornerstone and first step in establishing the breeding program, the present findings could be a basis for development of such programs.

Translational Potential of Baicalein in Mitigating RSL3-Induced Ferroptosis in Fibroblasts: Implications for Therapeutic Interventions.

Background: Ferroptosis is an iron-driven cell-death mechanism that plays a central role in various diseases. Recent studies have suggested that baicalein inhibits ferroptosis, making it a promising therapeutic candidate. Materials and Methods: Fibroblast cultures were treated with different agents to determine the effects of baicalein on ferroptosis. Ferroptosis-related gene expression, lipid peroxidation, and post-treatment cellular structural changes were measured using real-time quantitative polymerase chain reaction, C11-BODIPY dye, and transmission electron microscopy, respectively. Results: Baicalein significantly inhibited rat sarcoma virus selective lethal 3-induced ferroptosis in fibroblasts. Moreover, in baicalein-treated groups, reduced ferroptosis-related gene expression, decreased lipid peroxidation, and maintained cell structure was observed when compared with those of the controls. Discussion: The ability of baicalein to counteract RSL3-induced ferroptosis underscores its potential protective effects, especially in diseases characterized by oxidative stress and iron overload in fibroblasts. Conclusion: Baicalein may serve as a potent therapeutic agent against conditions in which ferroptosis is harmful. The compound's efficacy in halting RSL3-triggered ferroptosis in fibroblasts paves the way for further in vivo experiments and clinical trials.

Identifying the microstructure and mechanical properties of expansive soil stabilized using fly ash and waste foundry sand

Expansive soil causes road construction damage in the form of cracks or deformations due to changes in moisture content. One method to address this issue is soil stabilization using additive materials. Special additives can sometimes be expensive; hence it is necessary to use additive materials based on industrial waste as a waste management solution to solve the problem of expansive soil as a road construction subgrade. This study aims to analyze the effects of adding two types of industrial waste, namely fly ash (FA) and waste foundry sand (WFS), regarding the microstructure, physical, and mechanical properties of expansive soil. The method of stabilizing expansive soil involves mixing the soil with 9 % FA and various levels of WFS (0 %, 7.5 %, 10 %, 15 %) based on the dry weight of the soil. Microstructure testing includes quantitative X-ray diffraction and scanning electron microscopy. Physical property testing includes specific gravity, Atterberg limits and sieve analysis. Mechanical testing, i.e. compaction, California Bearing Ratio, swelling, and Triaxial tests were conducted. Soil stabilized with 9 % FA and 15 % WFS shows a significant increase in the internal friction angle and cohesion, reducing swelling by 67.18 % compared to the original soil swelling. The addition of 9 % fly ash and 15 % WFS to expansive soil reduces the content of montmorillonite. Natural expansive soils have a very poor soaked CBR of 0.94 %, while stabilized soils with 9 % FA and 15 % WFS have a soaked CBR of 6.46 %. This means that the soaked CBR of the stabilized soil meets the minimum CBR required for road subgrade construction. A mixture of 9 % FA and 15 % WFS in expansive soil can be recommended as a material for stabilizing expansive soil due to its ability to improve microstructure and mechanical properties

Atom counting based on Voronoi averaged STEM intensities using a crosstalk correction scheme

If quantitative scanning transmission electron microscopy is used for very precise thickness measurements with atomic resolution, it is commonly referred to as »atom counting«. Due to scattering and the finite probe extent the signal recorded in one atomic column is dependent not only on its own height but also on the height of its neighbours. Especially for thicker specimens this crosstalk effect can have significant impact on the measured intensity. If it is not appropriately accounted for in the evaluation, it can result in a deterioration of accuracy that impedes the possibility of actual atom counting. However, as the number of possible neighbour configurations can be excessively large, a comprehensive consideration of all in the evaluation reference is neigh impossible. This work proposes a method that allows for the a-posteriori reduction of crosstalk during the evaluation by algebraic means. Based on a parametric model, which is described in detail in the article, the crosstalk is expressed by an invertible matrix. Applying the inverted matrix to the measurement yields crosstalk corrected intensity values with very little computational effort. These can subsequently be evaluated by direct comparison to simple reference data. The working principle of the method is presented on the example of crystalline gold. The crosstalk parametrisation is found by fitting a model to sets of specifically created multislice simulations. The parameters are given for both aberration corrected and uncorrected STEM. Subsequently the abilities and potential of the technique are assessed in simulative studies on multiple model systems including gold nanoparticles. Overall a significant and robust improvement of the attainable precision can be demonstrated making the proposed method a promising tool for reference-based atom counting.

Cell surface plasticity in response to shape change in the whole organism

Plasma membrane rupture can result in catastrophic cell death. The skeletal muscle fiber plasma membrane, the sarcolemma, provides an extreme example of a membrane subject to mechanical stress since these cells specifically evolved to generate contraction and movement. A quantitative model correlating ultrastructural remodeling of surface architecture with tissue changes invivo is required to understand how membrane domains contribute to the shape changes associated with tissue deformation in whole animals. We and others have shown that loss of caveolae, small invaginations of the plasma membrane particularly prevalent in the muscle sarcolemma, renders the plasma membrane more susceptible to rupture during stretch.1,2,3 While it is thought that caveolae are able to flatten and be absorbed into the bulk membrane to buffer local membrane expansion, a direct demonstration of this model invivo has been unachievable since it would require measurement of caveolae at the nanoscale combined with detailed whole-animal morphometrics under conditions of perturbation. Here, we describe the development and application of the "active trapping model" where embryonic zebrafish are immobilized in a curved state that mimics natural body axis curvature during an escape response. The model is amenable to multiscale, multimodal imaging including high-resolution whole-animal three-dimensional quantitative electron microscopy. Using the active trapping model, we demonstrate the essential role of caveolae in maintaining sarcolemmal integrity and quantify the specific contribution of caveolar-derived membrane to surface expansion. We show that caveolae directly contribute to an increase in plasma membrane surface area under physiologically relevant membrane deformation conditions.

Defining Mitochondrial Cristae Morphology Changes Induced by Aging in Brown Adipose Tissue.

Mitochondria are required for energy production and even give brown adipose tissue (BAT) its characteristic color due to their high iron content and abundance. The physiological function and bioenergetic capacity of mitochondria are connected to the structure, folding, and organization of its inner-membrane cristae. During the aging process, mitochondrial dysfunction is observed, and the regulatory balance of mitochondrial dynamics is often disrupted, leading to increased mitochondrial fragmentation in aging cells. Therefore, it is hypothesized that significant morphological changes in BAT mitochondria and cristae will be present with aging. A quantitative 3D electron microscopy approach is developed to map cristae network organization in mouse BAT to test this hypothesis. Using this methodology, the 3D morphology of mitochondrial cristae is investigated in adult (3-month) and aged (2-year) murine BAT tissue via serial block face-scanning electron microscopy (SBF-SEM) and 3D reconstruction software for manual segmentation, analysis, and quantification. Upon investigation, an increase is found in mitochondrial volume, surface area, and complexity and decreased sphericity in aged BAT, alongside significant decreases in cristae volume, area, perimeter, and score. Overall, these data define the nature of the mitochondrial structure in murine BAT across aging.

Quantitative and functional characterisation of extracellular vesicles after passive loading with hydrophobic or cholesterol-tagged small molecules

Extracellular vesicles (EVs) are nanosized intercellular messengers that bear enormous application potential as biological drug delivery vehicles. Much progress has been made for loading or decorating EVs with proteins, peptides or RNAs using genetically engineered donor cells, but post-isolation loading with synthetic drugs and using EVs from natural sources remains challenging. In particular, quantitative and unambiguous data assessing whether and how small molecules associate with EVs versus other components in the samples are still lacking. Here we describe the systematic and quantitative characterisation of passive EV loading with small molecules based on hydrophobic interactions - either through direct adsorption of hydrophobic compounds, or by membrane anchoring of hydrophilic ligands via cholesterol tags. As revealed by single vesicle imaging, both ligand types bind to CD63 positive EVs (exosomes), however also non-specifically to other vesicles, particles, and serum proteins. The hydrophobic compounds Curcumin and Terbinafine aggregate on EVs with no apparent saturation up to 106–107 molecules per vesicle as quantified by liquid chromatography – high resolution mass spectrometry (LC-HRMS). For both compounds, high density EV loading resulted in the formation of a population of large, electron-dense vesicles as detected by quantitative cryo-transmission electron microscopy (TEM), a reduced EV cell uptake and a toxic gain of function for Curcumin-EVs. In contrast, cholesterol tagging of a hydrophilic mdm2-targeted cyclic peptide saturated at densities of ca 104–105 molecules per vesicle, with lipidomics showing addition to, rather than replacement of endogenous cholesterol. Cholesterol anchored ligands did not change the EVs' size or morphology, and such EVs retained their cell uptake activity without inducing cell toxicity. However, the cholesterol-anchored ligands were rapidly shed from the vesicles in presence of serum. Based on these data, we conclude that (1) both methods allow loading of EVs with small molecules but are prone to unspecific compound binding or redistribution to other components if present in the sample, (2) cholesterol anchoring needs substantial optimization of formulation stability for in vivo applications, whereas (3) careful titration of loading densities is warranted when relying on hydrophobic interactions of EVs with hydrophobic compounds to mitigate changes in physicochemical properties, loss of EV function and potential cell toxicity.

Impact of wet-dry cycles on enhanced rock weathering of brucite, wollastonite, serpentinite and kimberlite: Implications for carbon verification

Enhanced rock weathering is a proposed CO2 removal strategy for mitigating climate change; its implementation can be facilitated by improving carbon verification methods. In this study, weathering experiments exposed brucite, wollastonite skarn, serpentinite, and kimberlite residues (Venetia Diamond Mine, South Africa), to wetting and drying cycles (4/day) for 1 yr to elucidate reaction pathways and rates while evaluating carbon verification tools, including mineral quantification, total inorganic carbon (TIC), and stable and radiogenic carbon isotopes. Two primary reaction pathways were identified: A) silicate and hydroxide dissolution leading to carbonate precipitation (desirable), and B) dissolution and reprecipitation of carbonates (undesirable). The kimberlite residues, containing serpentine (19–33 wt%) and calcite (4–5 wt%), experienced variable and minor changes in total inorganic carbon. The δ13C and δ18O values of the carbonate minerals approached those expected for atmospheric-derived CO2 and are best explained by the exchange of carbonate CO2 with atmospheric CO2 (pathway A) as opposed to net CO2 sequestration. In contrast, the reaction of brucite (1.22% to 5.98%) and wollastonite skarn (0.22% to 1.01%) with atmospheric CO2 was substantial, as indicated by the increases in TIC (pathway B) yet decreases in δ13C values were inconsistent with the incorporation of atmospheric CO2 as a result of kinetic isotope fractionation due to carbonation being CO2 supply limited. The pulverized serpentinite experience insufficient carbonation to be detected by TIC. Unweathered (blue ground) and naturally weathered (yellow ground) kimberlite from the Voorspoed Diamond Mine, South Africa were compared to kimberlite residues used in experiments. Yellow ground contained twice the TIC content compared to blue ground whereas quantitative scanning electron microscopy revealed calcite veinlets that are characteristic of secondary mineralization. While the blue and yellow ground samples were essentially radiocarbon dead (0.03 and 0.05 F14C, respectively), substantial modern carbon was incorporated into the kimberlite (up to +0.27 F14C; CO2 exchange), brucite (+0.55 F14C), and wollastonite skarn (+0.53 F14C; CO2 sequestration) powders during weathering experiments. Although stable and radiogenic carbon isotopes have been used to verify the mineralization of atmospheric CO2, these methods are affected by CO2 exchange and kinetic fractionation effects during weathering that can be misleading and, thus, should not be relied on as sole methods for carbon verification.

Effects of small pre-stretching on the microstructural evolution and strengthening of an Al–Cu–Mg–Ag alloy

Pre-stretching prior to aging is known to degrade the performance of Al–Cu–Mg–Ag alloys at elevated temperatures. In this work, small pre-stretching by 0.5% and 1.8% is selected to explore its potential influences on the microstructure and strengthening capacity of an Al–Cu–Mg–Ag alloy. The strength and microstructure of Al–Cu–Mg–Ag alloy are found to be sensitive to the pre-stretching. Even 0.5% pre-stretching can reduce the alloy strength by more than 10 MPa. Quantitative transmission electron microscopy (TEM) and atom probe tomography (APT) analysis indicate that this detrimental effect of pre-stretching on strength is directly associated with the suppressed Mg–Ag co-clustering, which greatly delays the Ω precipitation and induces a weaker strengthening effect. Quantitative calculation reveals that Mg–Ag co-clusters exhibit the greatest strengthening contribution to the yield strength of the underaged material because of its strongest cluster hardening, which is mainly originated from its order hardening capacity. As the pre-stretching increases from 0.5% to 1.8%, the θ coarsening is progressively intensified during exposing at 200 °C, accompanied by a gradual drop in the number density of Ω phase. Besides, the formation of a cubic σ phase is found to be considerably enhanced after exposing at 200 °C for 500h as the pre-stretching increases.

Specialized Cellular Domains for ABCA1-mediated Cholesterol Efflux Detected by Quantitative Electron Microscopy.

Specialized Cellular Domains for ABCA1-mediated Cholesterol Efflux Detected by Quantitative Electron Microscopy.

Maximizing Modalities: Accelerating Quantitative Multimodal Electron Microscopy.

Maximizing Modalities: Accelerating Quantitative Multimodal Electron Microscopy.

The dynamics and removal efficiency of antibiotic resistance genes by UV-LED treatment: An integrated research on single- or dual-wavelength irradiation

Antimicrobial resistance has gained increasing attention, because of the awareness of its potential health risks. Strategies for the removal of antibiotic resistance genes (ARGs) are urgently required. In the present study, UV-LEDs at wavelength of 265 and 285 nm were integrated at five conditions, including single 265 nm UV-LED, single 285 nm UV-LED, and combined 265 nm and 285 nm UV-LED at different intensities, to remove tet A, cat 1, and amp C. The ARGs removal efficiency, gene behavior, and possible cellular mechanism were analyzed using real-time quantitative PCR, flow cytometry, and transmission electron microscopy (TEM). The 265 nm UV-LED is more effective than the 285 nm UV-LED and their combinations in terms of ARGs control, in which 1.91, 1.71, and 1.45 log were removed for tet A, cat 1, and amp C, respectively, at a UV dosage of 500 mJ/cm2. The intracellular gene leakage was detected in all five UV-LED experiment scenarios even when the cell membrane damage was insignificant with the highest increase of 0.69 log ARGs. ROS was generated during the irradiation, and the ROS was strongly negative correlated with intracellular ARGs, which could promote the degradation and removal of ARGs. This study provides a new insight of intracellular ARGs removal, because direct irradiation, ROS oxidation, and leakage to the extracellular serve as the three main pathways under high-dosage UV-LED irradiation. Further research should be focused on the mechanism and optimization of UV technology with 265 nm UV-LED for ARG control.

Selective autophagy associated with iron overload aggravates non-alcoholic steatohepatitis via ferroptosis.

Non-alcoholic steatohepatitis (NASH) is a progressive form of non-alcoholic fatty liver disease (NAFLD) that causes cirrhosis and hepatocellular carcinoma. Iron is an essential trace element in the body; however, excess iron can cause tissue damage and dysfunction. Iron overload is often observed in patients with NASH, and the amount of iron accumulated in the liver positively correlates with the histological severity of NASH. Ferroptosis, a novel form of iron-dependent cell death, is caused by the accumulation of lipid peroxidation and oxidative stress and is related to NASH. In addition, ferroptosis is closely related to autophagy, an intracellular self-degradation process. Although autophagy has many beneficial effects, it may also be harmful to the organism, for example, inducing ferroptosis. It is unclear whether iron overload aggravates NASH via autophagy. The aim of this research is to determine the mechanism by which iron overload induces ferroptosis via autophagy and aggravates NASH. Stroke-prone spontaneously hypertensive rats (SHRSP5/Dmcr) were divided into two groups and fed a high-fat and high-cholesterol (HFC) diet for eight weeks. Iron dextran was administered to the Fe group in addition to the HFC diet. Blood analysis, histological staining, calcineurin activity assay, quantitative reverse transcription polymerase chain reaction (RT-PCR), immunofluorescence staining, and electron microscopy were performed. The results showed that iron overload promoted autophagy via nuclear translocation of transcription factor EB (TFEB) and induced ferritinophagy, which is the autophagic degradation of ferritin. In addition, the HFC diet induced lipophagy, the autophagic degradation of lipid droplets. The Fe group also exhibited promoted ferroptosis and aggravated hepatic inflammation and fibrosis. In conclusion, iron overload accelerates ferritinophagy and lipophagy, aggravating NASH pathology via ferroptosis. These findings indicate the therapeutic potential of inhibiting autophagy and ferroptosis for treating NASH.

Interpretation of Hyperspectral Shortwave Infrared Core Scanning Data Using SEM-Based Automated Mineralogy: A Machine Learning Approach

Understanding the mineralogy and geochemistry of the subsurface is key when assessing and exploring for mineral deposits. To achieve this goal, rapid acquisition and accurate interpretation of drill core data are essential. Hyperspectral shortwave infrared imaging is a rapid and non-destructive analytical method widely used in the minerals industry to map minerals with diagnostic features in core samples. In this paper, we present an automated method to interpret hyperspectral shortwave infrared data on drill core to decipher major felsic rock-forming minerals using supervised machine learning techniques for processing, masking, and extracting mineralogical and textural information. This study utilizes a co-registered training dataset that integrates hyperspectral data with quantitative scanning electron microscopy data instead of spectrum matching using a spectral library. Our methodology overcomes previous limitations in hyperspectral data interpretation for the full mineralogy (i.e., quartz and feldspar) caused by the need to identify spectral features of minerals; in particular, it detects the presence of minerals that are considered invisible in traditional shortwave infrared hyperspectral analysis.