Transmission Electron Microscopy Research Articles

Novel Bacteriophage KG853 Exhibits Potent Lytic Activity and Biofilm Inhibition Against Pseudomonas aeruginosa

This study reports the isolation and characterization of a novel bacteriophage, KG853, specifically targeting Pseudomonas aeruginosa ATCC 27853. Morphological analysis using transmission electron microscopy revealed that bacteriophage KG853 belongs to the Bruynoghevirus genus. The phage demonstrated favorable characteristics for potential therapeutic applications, including a short latent period of 30 minutes and a large burst size of 136 plaque-forming units (PFU) per cell. KG853 exhibited stability across various temperatures and pH values, indicating its robustness under various environmental conditions. Genomic analysis showed that KG853 possesses a circular DNA genome of 45,390 base pairs with a GC content of 52.2%. No lysogenic or virulence genes were detected among the 84 open reading frames annotated in the genome, suggesting its safety for potential therapeutic use. Phylogenetic analysis revealed that phage KG853 is closely related to phage PaP3. Notably, KG853 demonstrated the ability to inhibit the formation of 4-hour biofilms by P. aeruginosa, a critical virulence factor in many infections. Host range analysis showed that KG853 is specific to P. aeruginosa, an important characteristic for targeted therapy. These findings suggest that bacteriophage KG853 represents a promising candidate for combating drug-resistant P. aeruginosa infections. Its specific host range, robust physical characteristics, lack of harmful genes, and anti-biofilm activity make it a potential alternative to conventional antibiotics. Further research is warranted to explore its efficacy in in vivo models and potential clinical applications.

Real-time four-dimensional scanning transmission electron microscopy through sparse sampling

Abstract Four-dimensional scanning transmission electron microscopy (4-D STEM) is a state-of-theart image acquisition mode used to reveal high and low mass elements at atomic resolution. The acquisition of the electron momenta at each real space probe location allows for various analyses to be performed from a single dataset, including virtual imaging, electric field analysis, as well as analytical or iterative extraction of the object induced phase shift. However, the limiting factor in 4-D STEM is the speed of acquisition which is bottlenecked by the read-out speed of the camera, which must capture a convergent beam electron diffraction (CBED) pattern at each probe position in the scan. Recent developments in sparse sampling and image inpainting (a branch of compressive sensing) for STEM have allowed for real-time recovery of sparsely acquired data from fixed monolithic detectors, Further developments in compressive sensing for 4-D STEM have also demonstrated that acquisition speeds can be increased i.e. live video rate 4D imaging is now possible. In this work, we demonstrate the first practical implementations of compressive 4-D STEM for real-time inference on two different scanning transmission electron microscopes.

Effect of dietary glycerol monolaurate supplementation on gene expression, intestinal mucosa and microbiota of juvenile black sea bream (Acanthopagrus schlegelii).

Glycerol monolaurate (GML) is a chemical compound derived from the combination of lauric acid and glycerol, exhibiting potent antimicrobial properties. An eight-week feeding trial was conducted to evaluate the effects of dietary supplementation of Glycerol Monolaurate (GML) on juvenile black sea bream. The control diet included 24% fish meal, while five additional diets were formulated with increasing GML concentrations: 0.01% (GML1), 0.02% (GML2), 0.04% (GML3), 0.08% (GML4), and 0.16% (GML5). Each diet was assigned to triplicate tanks. At the end of the experiment, serum immune and antioxidant in liver were non-significantly different among the groups. The relative gene expression of IGF-1 and GH in liver was statistically higher in the GML3 treated. The relative gene expression of IL-1, TGF, TNF-1, CCK, CLDN1, CASS-3 and GPR41 in intestine were statistically higher in the GML3 group. The photomicrographs of transmission electron microscope of the intestine were also studied. The GML supplemented groups had longer brush borders. The cells seemed to be joined by an intact junctional complex without visible intercellular spaces. The observed variations in phyla, class, and genus did not demonstrate statistical significance. Proteobacteria, Cyanobacteria, Firmicutes and Actinobacteria in the phylum, and Oxyphotobacteria, Bacteroidia, Holophagae and Negativicutes in the class, Brevundimonas, and Achromobacter in the genera were all substantially increased with higher levels of GML supplementation. Based on these results, it is suggested that glycerol monolaurate (GML) may serve as a potential dietary supplement for promoting different gene expression and intestinal health in black sea bream.

Application of Advanced Microscopy Techniques to The Characterization of Mixed Matrix Membranes.

Mixed matrix membranes (MMMs) have emerged as promising materials for various separation processes due to their tunable properties, enhanced separation performance and reproducibility. In this review paper, we provide a comprehensive overview of the methodologies, challenges, and applications associated with the characterization of MMMs using two advanced imaging techniques: Focused Ion Beam Scanning Electron Microscopy (FIB-SEM) and Transmission Electron Microscopy (TEM). We begin by outlining the principles and capabilities of FIB-SEM and TEM, emphasizing their suitability for studying the microstructure, morphology, and composition of MMMs at nanoscale resolution. Subsequently, we discuss the specific challenges and limitations encountered in the characterization of MMMs using these techniques, including sample preparation, image acquisition, and data interpretation. Furthermore, we review the diverse applications of FIB-SEM and TEM in elucidating the structure-property relationships of MMMs. Through illustrative examples, we highlight the valuable insights gained from these imaging techniques in optimizing MMMs for various separation applications. Finally, we propose future directions and emerging trends in MMM characterization, including the integration of lasers into FIB-SEM and in situ characterization techniques, to address current challenges and push the boundaries of MMM design and performance.

Depth profiling of implanted D in silicates: Contribution of solar wind protons to water in the Moon and terrestrial planets

The solar wind protons implanted in silicate material and combined with oxygen are considered crucial for forming OH/H O on the Moon and other airless bodies. This process may also have contributed to hydrogen delivery to planetary interiors through the accretion of micrometre-sized dust and planetesimals during early stages of the Solar System. This paper experimentally investigates the depth distribution of solar wind protons in silicate materials and explores the mechanisms that influence this profile. We simulated solar wind irradiation by implanting 3 keV D ions in three typical silicates (olivine, pyroxene, and plagioclase) at a fluence of sim 1.4 × 10 ions/cm . Fourier transform infrared spectroscopy was used to analyse chemical bond changes, while transmission electron microscopy (TEM) characterised microstructural modifications. Nanoscale secondary ion mass spectrometry (NanoSIMS) was employed to measure the D/ O ratio and determine the depth distribution of implanted deuterium. The newly produced OD band (at 2400–2800 cm –1 ) in the infrared spectrum reveals the formation of O–D bonds in the irradiated silicates. The TEM and NanoSIMS results suggest that over 73<!PCT!> of the implanted D accumulated in fully amorphous rims with a depth of 70 nm, while 25<!PCT!> extended inwards to sim 190 nanometres, resulting in partial amorphisation. The distribution of these deuterium particles is governed by the collision processes of the implanted particles, which involve factors such as initial energy loss, cascade collisions, and channelling effects. Furthermore, up to 2<!PCT!> of the total implanted D penetrated the intact lattice via diffusion, reaching depths ranging from hundreds of nanometres to several micrometres. Our results suggest that implanted solar wind protons can be retained in silicate interiors, which may significantly affect the hydrogen isotopic composition in extraterrestrial samples and imply an important source of hydrogen during the formation of terrestrial planets.

Lhx6 deficiency causes human embryonic palatal mesenchymal cell mitophagy dysfunction in cleft palate

BackgroundOverconsumption of retinoic acid (RA) or its analogues/derivatives has been linked to severe craniomaxillofacial malformations, such as cleft palate and midface hypoplasia. It has been noted that RA disturbed the proliferation and migration of embryonic palatal mesenchymal (EPM) cells in these malformations, yet the exact mechanisms underlying these disruptions remained unclear.MethodsA model of retinoic acid (RA)-induced cleft palate in fetal mice was successfully established. Histological alterations in the palate were evaluated using Hematoxylin and Eosin (H&E) staining and RNA in situ hybridization (RNAscope). Cellular proliferation levels were quantified via the Cell Counting Kit-8 (CCK-8) assay and EdU incorporation assay, while cell migration capabilities were investigated using wound healing and Transwell assays. Mitochondrial functions were assessed through Mito-Tracker fluorescence, mitochondrial reactive oxygen species (ROS) measurement, ATP level quantification, and mitochondrial DNA (mtDNA) copy number analysis. Differential gene expression and associated signaling pathways were identified through bioinformatics analysis. Alterations in the transcriptional and translational levels of Lhx6 and genes associated with mitophagy were quantified using quantitative PCR (qPCR) and Western blot analysis, respectively. Mitochondrial morphology and the mitochondrial autophagosomes within cells were examined through transmission electron microscopy (TEM).ResultsAbnormal palatal development in mice, along with impaired proliferation and migration of human embryonic palatal mesenchymal (HEPM) cells, was associated with RA affecting mitochondrial function and concomitant downregulation of Lhx6. Knockdown of Lhx6 in HEPM cells resulted in altered cell proliferation, migration, and mitochondrial function. Conversely, the aberrant mitochondrial function, proliferation, and migration observed in RA-induced HEPM cells were ameliorated by overexpression of Lhx6. Subsequent research demonstrated that Lhx6 ameliorated RA-induced dysfunction in HEPM cells by modulating PINK1/Parkin-mediated mitophagy, thereby activating the MAPK signaling pathways.ConclusionLhx6 is essential for mitochondrial homeostasis via tuning PINK1/Parkin-mediated mitophagy and MAPK signaling pathways. Downregulation of Lhx6 by RA transcriptionally disturbs the mitochondrial homeostasis, which in turn leads to the proliferation and migration defect in HEPM cells, ultimately causing the cleft palate.Graphical abstract

Aqueous phase transfer of near-infrared ZnCuInS2/ZnS quantum dots: Synthesis and characterization

Cadmium-free and NIR fluorescent QDs are promising candidates for bio-application. Thus, we present the synthesis of ternary ZnCuInS2/ZnS (ZCIS/ZnS) quantum dots (QDs) where the molar variation of Cu/Zn of the precursors was used to tune the optical and structural properties. QDs with Cu/Zn molar ratio of 2/1 passivated with ZnS exhibited the best optical properties. They showed dominant near-infrared photoluminescence (approx. 850 nm) and highest quantum yield (approx. 52 %, λexc = 500 nm). Therefore, they were further subject to modification to ensure their transfer to the aqueous phase and improve biocompatibility. For this, different functionalization approaches were used. The first method relied on encapsulation with polymers like PSMA (poly(styrene co-maleic anhydride)) and PMAO (poly(maleic anhydride-alt-1-octadecene) coupled with polyetheramine (JEFF; Jeffamine M-1000), and the second relied on hydrophilization with PMAO. Furthermore, we also applied a surface ligand exchange process using DHLA (dihydrolipoic acid) and polyethylene glycol (PEG)-appended DHLA. The comprehensive study indicated that ZnCuInS2/ZnS QDs functionalized with PMAO (ZnCuInS2/ZnS@PMO) exhibited the highest photoluminescence (PL QY) along with ensured high colloidal stability in aqueous media. Moreover, no noticeable deterioration of the photoluminescence profile was observed for all used functionalization approaches. However, a significant decrease in QY was observed for almost all functionalized QDs except those that were PMO-capped. The synthesized QDs were systematically characterized by transmission electron microscopy (TEM), powder X-ray diffraction (XRD), UV–Vis absorption spectroscopy, and fluorescence spectroscopy. Biological studies indicate that the obtained hydrophilic ZCIS QDs are biocompatible and localized intracellularly inside endosomes.

Hydrothermal Synthesis of Magnetic Cube‐Like Copper Ferrite Nanocrystallines for Reduction 4‐Nitrophenol

AbstractThe cube‐like copper ferrite (CuFe2O4) nanocrystallines are produced by the hydrothermal method without using any surfactant. X‐ray diffraction (XRD), transmission electron microscopy (TEM), and Fourier transform infrared spectroscopy (FT‐IR) are used to characterize the morphology, structure, and phase of the samples. CuFe2O4 nanocubes show the superior catalytic performance of reduction 4‐nitrophenol (4‐NP) to 4‐aminophenol (4‐AP). 20 mg CuFe2O4 nanocubes can totally convert 4‐NP to 4‐AP in 30 s at room temperature. Moreover, the reaction kinetics of 4‐NP catalytic reduction are also studied. Hence, this innovative CuFe2O4 nanocube has a lot of potential for organic pollutant destruction.

High‐temperature deformation and consolidation of polycrystalline α‐SiC by spark plasma sintering

AbstractThe main objective of this investigation is the consolidation and flexural strength of alpha silicon carbide ceramics produced without additives by spark plasma sintering. Using the design of the experiment method, we optimized temperature and dwell to achieve fully dense silicon carbide bulks. The consolidation process of silicon carbide was analyzed similarly to the creep of bulk ceramics, and it was determined that the activation energy for the densification process was 596 ± 39 kJ/mol, while the stress exponent n was below 2. Bulk additive‐free silicon carbide ceramics gradually increased flexural strength as the temperature rose to 2000°C. The flexural strength at 2000°C was influenced by the loading rate, and under 2.5 mm/min, it reached a maximum of 2.08 GPa. To explain this phenomenon, a deformation mechanisms map was created, indicating that diffusion creep is the most probable mechanism for the strain sensitivity of SiC at 2000°C. Transmission electron microscopy indicated a substantial rise in twin density within the α‐SiC grains at 2000°C, indicating the activation of a previously unreported self‐reinforcing mechanism.

Evaluating fructose content in poultry feed: electrochemical insights

AbstractThe global demand for poultry meat has increased, but consumers have become more picky in their nutritional choices. The fat content of poultry meat has risen since genetic advancement has resulted in higher rates of body weight gain and fat deposition, which are closely linked. The increase in further processing for fast-food poultry items has resulted in larger birds with more body fat. Usually, carbohydrates are used for easy weight gain in poultry. This study focuses on the electrochemical detection of fructose as a feed additive in poultry. The catalyst used was silver–silver oxide–zinc oxide (Ag–AgO–ZnO) and it was prepared by sol–gel method. Further, the surface morphology of the catalyst was analysed using various techniques, including X-ray diffraction, Fourier-transform infrared spectroscopy and transmission electron microscopy (XRD, FTIR and TEM). Cyclic voltammetry was carried out to find out the effectiveness of this catalyst in detecting fructose and our results revealed a sensitivity of 0.3 M with an electrochemical current of 1 mA cm−2, indicating the effectiveness of the chosen electrochemical approach. And further investigation was carried out to monitor effects of various parameters including scan rate, catalyst loading and fructose concentration. This research contributes valuable insights into assessing fructose levels in poultry feed, with potential implications for optimising nutritional formulations and enhancing overall poultry health. The electrochemical method demonstrates promise as a reliable tool for sensitively analysing fructose in feed additives. Precision and accuracy assessments further underscore the reliability of our electrochemical approach in differentiating fructose content within the poultry feed matrix. Graphical Abstract

Effect of CTAB and NaBH4 Solutions on Ti and Cu Nanoparticles Prepared by Pulsed Laser Ablation

The effect of different media, including cetyltrimethylammonium bromide (CTAB) and sodium borohydride (NaBH4), on the size, shape, optical absorption, and stability of titanium (Ti) and copper (Cu) nanoparticles (NPs) synthesized by laser ablation in liquid, was investigated. Ti and Cu NPs were fabricated by irradiating Ti and Cu targets using a Nd:YAG pulsed laser with a wavelength of 1064 nm, an energy of 500 mJ, repetition rate of 1 Hz, and a number of pulses of 1000. Results showed the direct effect of the type of surfactant on the size and size distribution of Ti and Cu NPs under the same laser ablation parameters. Morphological properties were described by field emission scanning electron microscope (FESEM) images, which revealed the formation of semispherical particles. The EDS results confirmed the spike-induced synthesis of Ti and Cu in CTAB and NaBH4 solutions. The transmission electron microscopy (TEM) images showed the nanostructures of Ti and Cu with spherical particles. The UV-vis absorption spectra showed that the absorption peak of Ti was almost constant at 289 nm, while the peak of Cu disappeared for both solutions. Zeta potential analysis showed that NPs prepared in CTAB solution were more stable than those in NaBH4 solution.

Recent Advances in Zinc Oxide Nanoparticles: Synthesis Methods, Characterization Techniques, and Emerging Applications

Abstract: Zinc oxide (ZnO) is an inorganic compound with unique physicochemical characteris-tics that make it versatile and suitable for various applications, especially in the form of nanoparti-cles (NPs). ZnO nanoparticles (ZnO NPs) exhibit distinct properties and are produced through diverse techniques, making them valuable for applications ranging from consumer goods to medi-cal and catalytic uses. The increasing popularity of ZnO NPs is driven by novel synthesis methods that allow for modification of chemical composition and control over size and shape, thereby en-hancing their properties and expanding their applications. The catalytic activity of ZnO NPs is influenced by parameters such as oxophilicity, large surface area, amphoteric nature, and the zinc cation's ability to approach activated starting material supports, making them viable heterogeneous catalysts for a variety of applications. Various analytical techniques, including X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM), transmission electron microscopy (TEM) analysis, atomic force microscopy (AFM), and many more, are used to characterize the nanoparticles. This article explores various synthesis methods and characterization techniques and focuses on the catalytic activities of ZnO NPs

Synthesis of tin selenide nanoparticles using Polygonum avicular extract decorated on graphitic carbon nitride for enhancing photodegradation of amoxicillin trihydrate and photoproduction of hydrogen peroxide

In this research, tin selenide nanoparticles (SnSe-NPs) were synthesized using Polygonum avicular extract via the hydrothermal method and decorated on graphitic carbon nitride (g-C3N4) by the simple sonicated method to produce SnSe/g-C3N4 composites. The characterization and properties of the SnSe/g-C3N4 composites, as compared to pristine SnSe and g-C3N4, were determined by Fourier-transform infrared spectroscopy, X-ray powder diffraction, scanning electron microscope, transmission electron microscopy, field emission scanning electron microscopy, X-ray photoelectron spectroscopy, ultraviolet–visible diffuse reflectance spectroscopy, Mott-Schottky, and electrochemical impedance spectroscopy. As a result, the SnSe-NPs possessed the rod-like shape and after the decoration onto the g-C3N4 sheet surface, it reduced the bandgap energy of the g-C3N4 from 2.58 downward 1.43 eV. Besides, the photoactivities of the 10-SnSe/g-C3N4 catalyst were investigated via both sides of amoxicillin trihydrate (AMXT) photodegradation and hydrogen peroxide (H2O2) photoproduction, in which the results were achieved 87.84 % AMXT and 85.48 μM H2O2, respectively, under visible light. Furthermore, the photodegradation was consistent with the pseudo-first-order kinetics and it performed the highest photodegradation at pH ∼5, while the importance of isopropyl alcohol as a trapping agent and •O2− radicals was affirmed in the H2O2 photoproduction. Moreover, the photoproduction of H2O2 was remained after 4 cycles achieved at 62.70 % in the final cycle, which demonstrated the stability and reusability of the 10-SnSe/g-C3N4. These aforementioned results demonstrate that the SnSe/g-C3N4 material is a promising candidate for addressing environmental pollution and energy scarcity issues.

Design and Synthesis of Fluorine-Containing Embedded Carbon Dots Stationary Phase for Separation of Versatile Analytes.

Thus far, numerous new stationary phases have been developed. A fluorine-containing embedded carbon dots (F3-CDs-SiO2) stationary phase was first designed and synthesized. The resulting F3-CDs-SiO2 stationary phase was characterized carefully by scanning electron microscopy, transmission electron microscopy, elemental analysis, X-ray photoelectron spectroscopy, and Brunauer-Emmett-Teller experiment. The F3-CDs-SiO2 stationary phase was slurry packed into the HPLC column (150 × 2.1 mm) for evaluation. Furthermore, the F3-CDs-SiO2 column was successfully used for separation of pesticides, nucleosides, sulfonamides, alkaloids, and alkylbenzenes. The retention mechanism (including hydrophobic interaction, F-F, hydrogen bond interaction, ion-exchange, dipole-dipole interaction, electrostatic interaction, etc.) was investigated carefully. Meanwhile, the F3-SiO2 stationary phase was synthesized and used to evaluate the role of CDs. Furthermore, various commercial stationary phases (including amino-SiO2, diol-SiO2, C18-SiO2, and PFP-SiO2) were used for comparison. Moreover, the F3-CDs-SiO2 column possessed good repeatability, reproducibility, and stability in separation of versatile analytes.

The synergistic effect of α-tocopherol and phloretin-loaded nanoemulsions on improvement of the stability, antioxidant, and tyrosinase inhibitory potentiality.

The purpose of this study was to prepare and evaluate the formulation of nanoemulsions (NEs) to encapsulate phloretin (PT) to improve its stability, antioxidant, and tyrosinase inhibitory competence. The aim of this study was to improve the stability, antioxidant, and tyrosinase inhibitory effects of PT via NEs. The formulations were prepared using low energy emulsification method for PT-VE-NEs, α-tocopherol (Vitamin E) and medium chain triglycerides (MCT) were used as the oil phase, and Tween 60 was used as the emulsifier and PEG-400 as the co-emulsifier. The droplet size and zeta potential of oil-in-water NEs were evaluated using dynamic light scattering. The PT-VE-NEs were also characterized by transmission electron microscopy and Fourier transform infrared spectroscopy. The mean droplet diameter was 14.85±0.14nm, with a zeta potential of -2.47±0.51mV. Fourier transform infrared spectroscopy revealed the formation of molecular interactions in the NEs formulations. PT-VE-NEs size was maintained the same during the in vitro digestion study. The particle size of PT-VE-NE remained stable during in vitro digestion. The addition of VE significantly improved the antioxidant, tyrosinase inhibitory effects, as well as thelion and physical stability of PT-VE-NE. The results revealed that NEs is a promising strategy to improve the functionality and stability of PT and VE. PT-VE-NEs will be applied for the preservation of fruits.

The Synthesis and Application of Chitosan@Ag2O@Mn3O4 Three‐Phase Nanocomposite as a Heterogeneous Catalyst in the Synthesis of 2‐Hydroxynaphthalenepyrimidine Trione

ABSTRACTThe catalyst is often the most expensive element in a chemical process, rather than the reactants or products. Apart from the financial benefits, it is equally crucial to recover and reuse the catalyst to prevent waste and enhance the process's greenness. This study successfully synthesized the new, superior, and recyclable heterogeneous chitosan@Ag2O@Mn3O4 three‐phase nanocatalyst with high surface area and stability using Ag2O and Mn3O4 nanoparticles that stabilized on chitosan by copreparation method and was characterized by Fourier transform infrared (FT‐IR), scanning electron microscopy (SEM), energy dispersive X‐ray (EDX), transmission electron microscopy (TEM), surface mapping (mapping), X‐ray diffraction (XRD), Brunner–Emmett–Teller (BET), thermal gravimetric analysis (TGA), and differential scanning calorimetry (DSC) techniques in detail. Following, the efficiency of the synthesized nanocatalyst in the preparation of 2‐hydroxynaphthalenepyrimidine triones using the reaction of aromatic aldehydes, β‐naphthol, and barbituric acid derivatives was investigated and proved. The advantages of this method include performing reactions at room temperature, green media, short reaction times, and the easy purification of the products.

Bortezomib (BOR)‐Pegylated‐Gold Nanovector: Synthesis, Spectroscopic Evaluation and Diagnostic Tool as Galectin‐1 Biomarker

ABSTRACTIn this paper, we applied an original chemical methodology in which gold salt (HAuCl4) interacts with the chemotherapeutic drug (bortezomib; i.e., BOR) by chelation and then stacked with dicarboxylic acid‐terminated polyethylene‐glycol (PEG‐diacide) as a biocompatible surfactant. The suggested chemical protocol is rapid (“one‐pot”) and reproducible, providing the formation of a hybrid‐nanovector named BOR IN PEG‐AuNPs. In order to prove a therapeutic approach, our hybrid‐nanovector (BOR IN PEG‐AuNPs) interacts with Galectin‐1 (Gal‐1) protein biomarker under specific concentrations. The efficient concentration range of this nanovector is obviously profiled by tumor microenvironment (TME) heterogeneity, optimizing cells access to the interaction region. Considering several influential factors related to spatial mapping and physical profile in all extracellular matrix (ECM), drive a change in neighborhood electrical potential configuration, leading the nanovector response with biomarkers transcriptions, hence, patterning TME leads to promote antitumor immunity in favor of tumor suppression. Each step of chemical synthesis and detection was monitored by spectroscopic techniques (Raman; UV‐Vis spectroscopies) and transmission electron microscopy (TEM). Our study demonstrated that hybrid‐nanoparticle system represents a key to further synergic chemotherapeutic and diagnostic tools for the treatment of cancer.

Isolation and Characterization of Exosomes from Ocular Samples.

Exosomes are microsize vesicles secreted by nearly all cells to the extracellular space. The vesicles transport cell signaling and communicate with other cells. Ultracentrifugation is the standard method to isolate exosomes from culture media or body fluid. Without ultracentrifuge, exosomes can be precipitated by polyethylene glycol or separated by size exclusion chromatography. After isolation, nanoparticle tracking analysis can help to estimate the size and concentration of exosome samples. Transmission electron microscopy can directly show the size and morphology of exosomes. Moreover, the sample should be characterized by the expression of several exosome biomarker proteins. Exosomal contents such as proteins and miRNAs could be profiled using appropriate technologies.

Multiplex antimicrobial activities of the self-assembled amphiphilic polypeptide β nanofiber KF-5 against vaginal pathogens

BackgroundVaginal infections caused by multidrug-resistant pathogens such as Candida albicans and Gardnerella spp. represent a significant health challenge. Current treatments often fail because of resistance and toxicity. This study aimed to synthesize and characterize a novel amphiphilic polypeptide, KF-5, and evaluate its antibacterial and antifungal activities, biocompatibility, and potential mechanisms of action.ResultsThe KF-5 peptide was synthesized via solid-phase peptide synthesis and self-assembled into nanostructures with filamentous and hydrogel-like configurations. Characterization by scanning electron microscopy (SEM), transmission electron microscopy (TEM), and atomic force microscopy (AFM) confirmed the unique nanostructural properties of KF-5. KF-5 (125, 250, or 500 µg/ml) demonstrated potent antibacterial and antifungal activities, with significant inhibitory effects on drug-resistant Candida albicans and Gardnerella spp. (P < 0.05). In vitro assays revealed that 500 µg/ml KF-5 disrupted microbial cell membranes, increased membrane permeability, and induced lipid oxidation, leading to cell death (P < 0.05). Cytotoxicity tests revealed minimal toxicity in human vaginal epithelial cells, keratinocytes, and macrophages, with over 95% viability at high concentrations. Molecular dynamics simulations indicated that KF-5 interacts with phospholipid bilayers through electrostatic interactions, causing membrane disruption. In vivo studies using a mouse model of vaginal infection revealed that 0.5, 1, and 2 mg/ml KF-5 significantly reduced fungal burden and inflammation, and histological analysis confirmed the restoration of vaginal mucosal integrity (P < 0.01). Compared with conventional antifungal treatments such as miconazole, KF-5 exhibited superior efficacy (P < 0.01).ConclusionsKF-5 demonstrates significant potential as a safe and effective antimicrobial agent for treating vaginal infections. Its ability to disrupt microbial membranes while maintaining biocompatibility with human cells highlights its potential for clinical application. These findings provide a foundation for further development of KF-5 as a therapeutic option for combating drug-resistant infections.

Water vapor oxidation of SiC layer of tristructural isotropic particles

AbstractTristructural isotropic (TRISO) fuel particles, developed for use in high‐temperature gas‐cooled nuclear reactors, can be subjected to oxidizing environments in off‐normal accident scenarios. In this study, surrogate TRISO fuel particles were oxidized at 1000–1350°C for 4 h in 20 kPa water vapor atmosphere balanced with ultrahigh‐purity helium gas. The oxide scale morphology and thickness were studied via scanning electron microscopy, focused ion beam, and transmission electron microscopy. The oxide thickness increased as the oxidation temperature was increased. Although the oxide scale at 1000°C was completely amorphous, pockets of crystalline oxide were observed on particles oxidized at 1200 and 1300°C. Kinetic analysis was performed to deduce the oxidation mechanism by determining the apparent activation energy using linear regression analysis. The oxidation mechanism was consistent for temperatures from 1000 to 1200°C with an activation energy of 412.4 ± 8.8 kJ/mol. The activation energy then dropped to approximately 201 ± 7.1 kJ/mol for temperatures 1200–1350°C. Therefore, there is a change in oxidation mechanism at ∼1200°C. This change is attributed to the existence of a network of significant bubbles in the oxide layer at higher temperatures, which serve as rapid diffusion pathways for the transport of oxidants from the surface to the SiC/SiO2 interface, instead of relying on the bulk diffusion through the SiO2 layer at lower temperatures where only small bubbles are present along the SiC/SiO2 interface.