Laser Scanning Microscopy Research Articles

Multi-scale tissue fluorescence mapping with fiber optic ultraviolet excitation and generative modeling

Cellular imaging of thick samples requires physical sectioning or laser scanning microscopy, which can be restrictive, involved, and generally incompatible with high-throughput requirements. We developed fiber optic microscopy with ultraviolet (UV) surface excitation (FUSE), a portable and quantitative fluorescence imaging platform for thick tissue that enabled quick sub-cellular imaging without thin sections. We substantially advanced prior UV excitation approaches with illumination engineering and computational methods. Optical fibers delivered <300nm light with directional control, enabling unprecedented 50× widefield imaging on thick tissue with sub-nuclear clarity, and 3D topography of surface microstructure. Probabilistic modeling of high-magnification images using our normalizing flow architecture FUSE-Flow (made freely available as open-source software) enhanced low-magnification imaging with measurable localized uncertainty via variational inference. Comprehensive validation comprised multi-scale fluorescence histology compared with standard H&E histology, and quantitative analyses of senescence, antibiotic toxicity, and nuclear DNA content in tissue models via efficient sampling of thick slices from entire murine organs up to 0.4×8×12mm and 1.3 million cells per surface. This technology addresses long-standing laboratory gaps in high-throughput studies for rapid cellular insights.

Effect of Repolishing on Color Stability, Translucency, and Surface Roughness of Aged Monochromatic Dental Composites

Abstract Objective This study aimed to test repolishing effect on color stability, translucency, and surface roughness of aged monochromatic dental composite in artificial saliva, tea, mouthwash, and coffee after 1 month, simulating 2.5 years of clinical use. Materials and Methods Omnichroma, a monochromatic dental composite, and Z250 XT a nanohybrid multishade dental composite were used in this study. Specimens (n = 80) from each dental composite were prepared to determine color stability, translucency parameter, and surface roughness after repolishing of aged specimens in artificial saliva, tea, mouthwash, and coffee for 1 month. Scanning electron microscope and laser scanning microscope were used to study surface topography after repolishing. Data was analyzed using analysis of variance, Scheffe test, and independent t-test. Results A significant difference between both dental composites after repolishing in terms of color stability and translucency parameter as that of Omnichroma was higher but sill color changes of Omnichroma were perceptible. There was no difference between both composites in regard to surface roughness after repolishing; however, values of both composites were above the accepted value of 0.2 µm and laser scanning microscope images confirmed these findings. Conclusions Repolishing did not enhance color and surface roughness of aged monochromatic dental composites.

Study of proanthocyanidin promotes osteogenic differentiation of human periodontal ligament stem cells through the transcription factor EB-induced autophagy-lysosome pathway

Objective: To investigate the mechanism of proanthocyanidin (PA) in regulating the osteogenic differentiation of human periodontal ligament stem cells (PDLSCs), and to explore the effects of PA on the expression and nuclear translocation of transcription factor EB (TFEB) and on the autophagy-lysosome pathway. Methods: PDLSCs were divided into control group and PA group, which were subjected to RNA sequencing analysis (RNA Seq) to detect differentially expressed genes. The osteogenic differentiation ability and autophagy level were observed by real-time fluorescence quantitative PCR (RT-qPCR) analysis, alkaline phosphatase (ALP) staining and transmission electron microscope (TEM), respectively. Scratch assay and Transwell assay were used to detect the migration ability of PDLSCs. Lysotracker and immunofluorescence staining were used to detect the biogenesis of lysosomes. The total protein expression of transcription factor EB (TFEB) as well as that in cytoplasm and nucleus were detected by Western blotting. Confocal laser scanning microscope (CLSM) was used to observe the nuclear translocation of TFEB. The PDLSCs were treated with small interfering RNA (siRNA) technology to knock down the expression levels of TFEB gene with or without PA treatment. Western blotting was used to analyze the expressions of autophagy-related proteins Beclin1 and microtubule-associated protein 1 light chain 3 (LC3B), as well as osteogenic-related proteins runt-related transcription factor 2 (RUNX2), ALP, and osteocalcin in PDLSCs. Results: Compared with the control group, the osteogenic-related and autophagy-related genes showed differential expression in PDLSCs after PA treatment (P<0.05). The mRNA expression levels of osteogenic-related genes RUNX2 (2.32±0.15) and collagen type Ⅰ alpha 1 (COL1α1) (1.80±0.18), as well as the autophagy related genes LC3B (1.87±0.08) and Beclin1 (1.63±0.08) were significantly increased in the PA group, compared with the control group (1.01±0.16, 1.00±0.10, 1.00±0.07, 1.00±0.06, respectively, all P<0.01). Compared with the control group, the PA group had higher ALP activity, and more autophagosomes and autophagolysosomes observed by TEM. PA promoted the migration of PDLSCs (P<0.05) and the increased number of lysosomes and the expression of lysosomal associated membrane protein 1 (LAMP1). In the PA group, the relative expression level of total TFEB protein (1.49±0.07) and the nuclear/cytoplasmic expression of TFEB protein (1.52±0.12) were significantly higher than the control group (1.00±0.11, 1.00±0.13, respectively) (t=6.43, P<0.01; t=5.07, P<0.01). The relative nuclear/cytoplasmic fluorescence intensity of TFEB in the PA group (0.79±0.09) was increased compared with the control group (0.11±0.08) (t=8.32, P<0.01). Knocking down TFEB significantly reduced the expression of TFEB (1.00±0.15 vs 0.64±0.04), LAMP1 (1.00±0.10 vs 0.69±0.09), Beclin1 (1.00±0.05 vs 0.60±0.05), and LC3B Ⅱ/Ⅰ (1.00±0.06 vs 0.73±0.07) in PDLSCs (P<0.05, P<0.05, P<0.01, P<0.01). When TFEB gene was knocked down, the expression levels of Beclin1 (1.05±0.11), LC3B Ⅱ/Ⅰ (1.02±0.09), RUNX2 (1.04±0.10), ALP (1.04±0.16), and osteocalcin (1.03±0.15) proteins were significantly decreased in the PA group compared with the pre-knockdown period (1.28±0.03, 1.44±0.11, 1.38±0.11, 1.62±0.11, 1.65±0.17, respectively) (P<0.05, P<0.01, P<0.05, P<0.01, and P<0.01, respectively). Conclusions: PA promotes the osteogenic differentiation of PDLSCs through inducing the expression and nuclear translocation of TFEB and activating the autophagy-lysosome pathway.

Spectroscopic analysis of the sum-frequency response of the carbon-hydrogen stretching modes in collagen type I.

We studied the origin of the vibrational signatures in the sum-frequency generation (SFG) spectrum of fibrillar collagen type I in the carbon-hydrogen stretching regime. For this purpose, we developed an all-reflective, laser-scanning SFG microscope with minimum chromatic aberrations and excellent retention of the polarization state of the incident beams. We performed detailed SFG measurements of aligned collagen fibers obtained from rat tail tendon, enabling the characterization of the magnitude and polarization-orientation dependence of individual tensor elements Xijk2 of collagen's nonlinear susceptibility. Using the three-dimensional atomic positions derived from published crystallographic data of collagen type I, we simulated its Xijk2 elements for the methylene stretching vibration and compared the predicted response with the experimental results. Our analysis revealed that the carbon-hydrogen stretching range of the SFG spectrum is dominated by symmetric stretching modes of methylene bridge groups on the pyrrolidine rings of the proline and hydroxyproline residues, giving rise to a dominant peak near 2942cm-1 and a shoulder at 2917cm-1. Weak asymmetric stretches of the methylene bridge group of glycine are observed in the region near 2870cm-1, whereas asymmetric CH2-stretching modes on the pyrrolidine rings are found in the 2980 to 3030cm-1 range. These findings help predict the protein's nonlinear optical properties from its crystal structure, thus establishing a connection between the protein structure and SFG spectroscopic measurements.

The effect of different irrigation activation methods on the penetration of the irrigation solution into the dentinal tubules in teeth with calcium hydroxide applied and oval-shaped canals: Confocal laser scanning microscope analysis.

This study aimed to evaluate the efficacy of different irrigation techniques including standard needle irrigation (SNI), passive ultrasonic irrigation (PUI), EndoActivator (EA), XP-Endo Finisher (XPF), photon-induced photoacoustic streaming (PIPS), and shock wave enhanced emission photo-acoustic streaming (SWEEPS) systems on the penetration of irrigation solutions into dentinal tubules in teeth with calcium hydroxide (CH) applied and oval-shaped canals by confocal laser scanning microscope (CLSM). Ninety mandibular incisor teeth with oval-shaped canals were included in this study. After the preparation of teeth using Resiproc Blue R25 (VDW, Munich, Germany) canals were filled with a paste based on CH. The teeth were stored in 100% humidity at a temperature of 37°C for 14 days. According to the irrigation activation systems, the teeth were divided into six groups (n = 15); SNI, PUI, EA, XPF, PIPS, and SWEEPS. In each group, 3 irrigation/activation cycles of 20 s of irrigation and 20 s of activation were performed with the relevant activation method. The root canals were filled with fluorescein sodium (Sigma, Alldrich Co., St. Louis, MO, USA) and activated with the relevant activation method for 30 s. Specimens were sectioned horizontally to 1 ± 0.1 mm at 2, 5, and 8 mm from the apex and then examined under the CLSM. Maximum penetration depth, maximum penetration area and penetration percentage were measured by using Image J software. Data were analyzed using a two-way analysis of variance (ANOVA) and posthoc Tukey tests (p = .05). No difference was found between the activation systems in terms of maximum penetration depth and maximum penetration area of irrigation solutions in the apical section (p > .05). The penetration percentage of irrigation solutions was higher in PUI and PIPS compared with EA and XPF at the apical section (p < .05). No difference was found between SNI, PUI, PIPS, and SWEEPS in terms of the penetration percentage of irrigation solutions in all sections (p > .05). The penetration percentage of irrigation solutions was higher in the middle and coronal sections compared to the apical section in EA and XPF (p < .05). Tubule penetration of irrigation solutions in SNI was similar between sections. The Tubule penetration area of irrigation solutions in PUI, EA, XPF, PIPS, and SWEEPS was lower in the apical section than in the middle and coronal sections. RESEARCH HIGHLIGHTS: The tubule penetration depth and area of the irrigation solutions were similar between the activation systems tested in the apical sections. The Tubule penetration area of irrigation solutions in middle and coronal sections was higher in PUI and PIPS than in SNI.

Comparative evaluation of different surface pretreatment methods on the depth of penetration of adhesive resin in sandwich technique: A confocal laser scanning microscopy study.

The aim of the study was to evaluate and compare the depth of penetration of adhesive resin in resin-modified glass ionomer cement (RMGIC) after different surface pretreatment methods using a confocal laser scanning microscope (CLSM). Class I cavity preparation was done in 45 freshly extracted premolars and divided into three groups, according to the surface pretreatment of RMGIC which was placed in a thickness of 1 mm in the cavity. In the control group, no pretreatment was done, and other two groups were pretreated with acid etching and air abrasion (AA). The fifth-generation dentin bonding agent mixed with rhodamine B dye 0.1% was applied on RMGIC in all the samples and cured for 15 s. Samples were restored using composite resin and sectioned longitudinally. The depth of penetration of adhesive resin was evaluated using CLSM. Depth of penetration of adhesive resin was highest with AA (153.70 ± 10.23), followed by acid etching (122.71 ± 12.25) and control group (77.12 ± 6.37). Based on the findings of this research, AA enhances the depth of penetration of adhesive resin in the RMGIC surface. Thus, AA before placement of composite resin on the RMGIC can be effective in a clinical scenario.

Natural product nobiletin-loaded Pickering emulsion stabilized by bovine serum albumin/carboxymethyl inulin complexes: preparation and digestive characteristics.

To expand the application of nobiletin (NOB) in semi-solid functional foods, bovine serum albumin (BSA)/carboxymethyl inulin (CMI) complexes-stabilized Pickering emulsion (BCPE) (φoil = 60%, v/v) was fabricated, and the swallowing index and bioavailability of the NOB-loaded Pickering emulsion was evaluated. Confocal laser scanning microscope (CLSM) and cryo-scanning electron microscopy (cryo-SEM) images revealed that BSA/CMI complexes attached to the oil-water interface. NOB-loaded BCPE exhibited a viscoelastic and shear-thinning behavior. Fork drip test results suggested that the textural value of unloaded and NOB-loaded emulsions was International Dysphagia Diet Standardisation Initiative Level 4, which could be swallowed directly without chewing. The in vitro lipolysis model suggested that NOB had a faster digestive profile and a higher bioaccessibility in the BCPE than in the oil suspension. The in vivo rat model revealed that the oral bioavailability of NOB was increased by 2.07 folds in BCPE compared to its bioavailability in unformulated oil. Moreover, BCPE led to a higher plasma concentration of the major demethylated metabolite of NOB (4'-demethylnobiletin) than the unformulated oil. Accordingly, BCPE enhanced the oral bioavailability of NOB by improving bioaccessibility, absorption, and biotransformation.

Influence of root canal moisture on the penetration of TotalFill bioceramic sealer into the dentinal tubules: A confocal laser scanning microscopy study

Abstract Introduction: Bioceramic (BC) sealers exhibit multiple desirable biologic and physical properties. The aim of this study was to evaluate the effect of root canal moisture on the depth of TotalFill BC sealer penetration into dentinal tubules at different levels using a confocal laser scanning microscope (CLSM). Materials and Methods: The manuscript of this laboratory study has been written according to preferred reporting items for laboratory studies in endodontology 2021 guidelines. Forty single-rooted extracted human premolars were decoronated using a low-speed diamond disc, followed by root canal preparation using nickel–titanium files under copious irrigation. The samples were randomly allocated for obturation into four groups n = 10 based on the sealer and root canal moisture status. Group A: TotalFill BC sealer in wet canal, Group B: TotalFill BC sealer in dry canal, Group C: AH Plus sealer in wet canal, and Group D: AH Plus sealer in dry canal. The roots were sectioned at three different distances from the apex. The root segments were examined using CLSM to measure the extent of sealer penetration into the dentinal tubules. The data underwent analysis through a three-way mixed ANOVA test, followed by post hoc assessments (P &lt; 0.05). Results: In wet canals, the tubular penetration of TotalFill BC sealer was notably greater than that of AH plus, with a statistically significant difference. However, AH Plus sealer had a statistically significant tubular penetration in the dry canals compared to the wet. Greater sealer penetration was observed at 5 mm from the apex, followed by 3 mm, and the least penetration depth was found at 1 mm from the apex. Conclusions: The presence of root canal moisture and distance from the apex had a significant impact on the penetration of TotalFill BC sealer into the dentinal tubules.

Purification, characterization, and identification of a novel bacteriocin produced by Lacticaseibacillus casei KLS1, and its antimicrobial mechanism against Staphylococcus aureus

Staphylococcus aureus (S. aureus) and its biofilm were recognized as one of the major threats to food safety and human health. Bacteriocins produced by Lacticaseibacillus species have been demonstrated to inhibit the growth of foodborne pathogenic bacteria and their biofilm formation. Herein, a novel bacteriocin KLS1 from Lacticaseibacillus casei KLS1 was extracted by ethyl acetate and purified using an AKTA Pure 25 protein chromatography purification system. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analyses revealed that the molecular mass of bacteriocin KLSI was 3786.91 Da and the amino acid sequence was DTSASIASNKSETNDLLKQIEAANTEVINLNKQID. Bacteriocin KLS1 was stable at pH 2.0–6.0, sensitive to proteases, and showed excellent thermal stability. Furthermore, bacteriocin KLS1 exhibited broad-spectrum antimicrobial activity against both Gram-positive and Gram-negative bacteria, even fungi. The crystal violet staining assay indicated that bacteriocin KLS1 significantly reduced the relative percentage of S. aureus biofilm (P < 0.05). Meanwhile, bacteriocin KLS1 was found to significantly decrease the metabolic activity of S. aureus by Cell Counting Kit-8 assay (P < 0.05). In addition, scanning electron microscopy and confocal laser scanning microscope showed that bacteriocin KLS1 disrupted the structure of S. aureus biofilm and cell membrane integrity, leading to cell deformation and leakage of contents. These results suggested that bacteriocin KLS1 might have the potential to serve as a biological preservative in the food industry.

Fangchinoline inhibits growth and biofilm of Candida albicans by inducing ROS overproduction.

Infections caused by Candida species, especially Candida albicans, threaten the public health and create economic burden. Shortage of antifungals and emergence of drug resistance call for new antifungal therapies while natural products were attractive sources for developing new drugs. In our study, fangchinoline, a bis-benzylisoquinoline alkaloid from Chinese herb Stephania tetrandra S. Moore, exerted antifungal effects on planktonic growth of several Candida species including C. albicans, with MIC no more than 50 μg/mL. In addition, results from microscopic, MTT and XTT reduction assays showed that fangchinoline had inhibitory activities against the multiple virulence factors of C. albicans, such as adhesion, hyphal growth and biofilm formation. Furthermore, this compound could also suppress the metabolic activity of preformed C. albicans biofilms. PI staining, followed by confocal laser scanning microscope (CLSM) analysis showed that fangchinoline can elevate permeability of cell membrane. DCFH-DA staining suggested its anti-Candida mechanism also involved overproduction of intracellular ROS, which was further confirmed by N-acetyl-cysteine rescue tests. Moreover, fangchinoline showed synergy with three antifungal drugs (amphotericin B, fluconazole and caspofungin), further indicating its potential use in treating C. albicans infections. Therefore, these results indicated that fangchinoline could be a potential candidate for developing anti-Candida therapies.

The influence mechanisms of re-fused scanning on the surface roughness, microstructural evolution and mechanical properties of laser powder bed fusion processed AlMgScZr alloy

Laser powder bed fusion (LPBF) stands out as the foremost method for achieving intricate structure with superior design flexibility among additive manufacturing technologies. Within the LPBF process, surface roughness serves as a critical metric governing both fabrication precision and build quality. This study delves into the evolution of the top and side surface roughness as re-fused scanning times increase in LPBF-fabricated AlMgScZr alloy. Concurrently, the investigation evaluates alterations in microstructural characteristics and mechanical properties. Various characterization techniques, including confocal laser scanning microscope, scanning electron microscope, X-ray diffraction, electron backscatter diffraction, as well as tensile and microhardness tests, are employed. The findings concerning the top surface reveal that the initial application of re-fused scanning contributes to enhanced surface roughness. This improvement stems from refined grain structures and induced lattice distortions, facilitated by rapid heat dissipation and thermal recycling within the solidified layer. Consequently, microhardness increases. However, introducing the subsequent re-fused scanning exacerbates surface roughness and instigates Marongoni flow instability, magnesium depletion, and alterations in lattice structures. Moreover, with increasing the re-fused times, there is a noticeable enhancement in preferential crystallite growth orientation along the (200)α-Al crystal plane. Regarding the side surface, escalating re-fused times correlate with deteriorating roughness and alterations in molten pool shape and size, attributable to successive energy input. Despite these surface variations, minimal differences are observed in yield strength, tensile strength, and elongation. In essence, this research offers valuable insights into selecting optimal scanning methodologies during the LPBF process, thus guiding improvements in fabrication precision and component quality.

Investigation of Mechanical Properties of Polymer-Infiltrated Tetrapodal Zinc Oxide in Different Variants.

The aim of this study was to evaluate the influence of weight ratio, the shape of the precursor particles, and the application of a phosphate-monomer-containing primer on the mechanical properties of polymer infiltrated ceramic networks (PICNs) using zinc oxide. Two different types of zinc oxide particles were used as precursors to produce zinc oxide networks by sintering, each with two different densities resulting in two different weight ratios of the PICNs. For each of these different networks, two subgroups were built: one involving the application of a phosphate-monomer-containing primer prior to the infiltration of Bis-GMA/TEGDMA and one without. Elastic modulus and flexural strength were determined by using the three-point bending test. Vertical substance loss determined by the chewing simulation was evaluated with a laser scanning microscope. There was a statistically significant influence of the type of precursor particles on the flexural strength and in some cases on the elastic modulus. The application of a primer lead to a significant increase in the flexural strength and in most cases also in the elastic modulus. A higher weight ratio of zinc oxide led to a significantly higher elastic modulus. Few statistically significant differences were found for the vertical substance loss. By varying the shape of the particles and the weight fraction of zinc oxide, the mechanical properties of the investigated PICN can be controlled. The use of a phosphate-monomer-containing primer strengthens the bond between the infiltrated polymer and the zinc oxide, thus increasing the strength of the composite.

Spinning Disk Confocal Microscopy for Optimized and Quantified Live Imaging of 3D Mitochondrial Network.

Mitochondria are the energy factories of a cell, and depending on the metabolic requirements, the mitochondrial morphology, quantity, and membrane potential in a cell change. These changes are frequently assessed using commercially available probes. In this study, we tested the suitability of three commercially available probes-namely 5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolo-carbocyanine iodide (JC-1), MitoTracker Red CMX Rox (CMXRos), and tetramethylrhodamine methyl ester (TMRM)-for assessing the mitochondrial quantity, morphology, and membrane potential in living human mesoangioblasts in 3D with confocal laser scanning microscope (CLSM) and scanning disk confocal microscope (SDCM). Using CLSM, JC-1, and CMXRos-but not TMRM-uncovered considerable background and variation. Using SDCM, the background signal only remained apparent for the JC-1 monomer. Repetitive imaging of CMXRos and JC-1-but not TMRM-demonstrated a 1.5-2-fold variation in signal intensity between cells using CLSM. The use of SDCM drastically reduced this variation. The slope of the relative signal intensity upon repetitive imaging using CLSM was lowest for TMRM (-0.03) and highest for CMXRos (0.16). Upon repetitive imaging using SDCM, the slope varied from 0 (CMXRos) to a maximum of -0.27 (JC-1 C1). Conclusively, our data show that TMRM staining outperformed JC-1 and CMXRos dyes in a (repetitive) 3D analysis of the entire mitochondrial quantity, morphology, and membrane potential in living cells.

Nucleation of Pitting and Evolution of Stripping on Lithium-Metal Anodes.

The stripping reaction of lithium (Li) will greatly impact the cyclability and safety of Li-metal batteries. However, Li pits' nucleation and growth, the origin of uneven stripping, are still poorly understood. In this study, we analyze the nucleation mechanism of Li pits and their morphology evolution with a large population and electrode area (>0.45 cm2). We elucidate the dependence of the pit size and density on the current density and overpotential, which are aligned with classical nucleation theory. With a confocal laser scanning microscope, we reveal the preferential stripping on certain crystal grains and a new stripping mode between pure pitting and stripping without pitting. Descriptors like circularity and the aspect ratio (R) of the pit radius to depth are used to quantify the evolution of Li pits in three dimensions. As the pits grow, growth predominates along the through-planedirection, surpassing the expanding rate in the in-plane direction. After analyzing more than 1000 pits at each condition, we validate that the overpotential is inversely related to the pit radius and exponentially related to the rate of nucleation. With this established nucleation-overpotential relationship, we can better understand and predict the evolution of the surface area and roughness of Li electrodes under different stripping conditions. The knowledge and methodology developed in this work will significantly benefit Li-metal batteries' charging/discharging profile design and the assessment of large-scale Li-metal foils.

Rheological Investigation of Highly Filled Copper(II) Oxide Nanosuspensions to Optimize Precursor Particle Content in Reductive Laser-Sintering

In this article, the particle concentration of finely dispersed copper(II) oxide nanosuspensions as precursors for reductive laser sintering (RLS) is optimized on the basis of rheological investigations. For this metallization process, a smooth, homogeneous and defect-free precursor layer is a prerequisite for adherent and reproducible copper structures. The knowledge of the rheological properties of an ink is crucial for the selection of a suitable coating technology as well as for the adjustment of the ink formulation. Different dilutions of the nanosuspension were examined for their rheological behavior by recording flow curves. A strong shear thinning behavior was found and the viscosity decreases exponentially with increasing dilution. The viscoelastic behavior was investigated by a simulated doctor blade coating process using three-interval thixotropy tests. An overshoot in viscosity is observed, which decreases with increasing thinning of the precursor. As a comparison to these results, doctor blade coating of planar glass and polymer substrates was performed to prepare precursor layers for reductive laser sintering. Surface morphology measurements of the resulting coatings using laser scanning microscopy and rheological tests show that homogeneous precursor layers with constant thickness can be produced at a particle–solvent ratio of 1.33. A too-high particle content results in an irregular coating layer with deep grooves and a peak-to-valley height Sz of up to 7.8 μm. Precise dilution control allows the fabrication of smooth surfaces with a Sz down to 1.5 μm.

Adsorption and inhibition of rosin thiourea imidazole quaternary ammonium salt on steel surface in HCl solution

Rosin thiourea imidazole quaternary ammonium salt (RTIQAs) was synthesized by connecting rosin and imidazole units via thiourea linker, and its inhibitive action on the corrosion of CRS (cold rolled steel) in 1.0 M HCl solution was fully investigated. The results demonstrate that RTIQAs is an efficient mixed-type inhibitor with the maximum inhibition efficiency of 95%. The adsorption rule on the CRS surface fully complies with Langmuir isotherm. The efficient inhibition of RTIQAs can also be verified by AFM, SEM and CLSM (confocal laser scanning microscope). The direct adsorption proof of RTIQAs molecules on steel surface can be further presented in EDX and XPS.

In situ study of microstructural evolution of borosilicate glass–sulfate mixture at high temperatures

AbstractMicrostructural evolution of borosilicate glass–sulfate (BaSO4 or Na2SO4) mixture was observed in situ using a high‐temperature confocal scanning laser microscope. The phase compositions and sulfate retention of the mixture at different temperatures were also examined. The results indicate that for the glass–BaSO4 sample (abbreviated as BS), the borosilicate glass begins to melt at about 900°C, and the triangular BaSO4 crystals grow obviously between 900°C and 1100°C. At temperatures above 1100°C, the BaSO4 crystals gradually melt and decompose. For the glass–Na2SO4 sample (abbreviated as NS), strip‐shaped BaSO4 crystals begin to form at about 820°C after the borosilicate glass melting, and then gradually melt with further increasing temperature. When the temperature increases to 1000°C, the upper layer of the NS sample possesses Na2SO4, LiNaSO4, and BaSO4 phases, while the lower layer possesses an amorphous phase. Moreover, the SO3 retention in the BS sample remains almost unchanged with temperature increases from 900°C to 1100°C, and then decreases with further increasing temperature. The SO3 retention retained in the NS sample decreases rapidly at temperatures above 900°C. These findings can provide a guidance for vitrifying sulfur‐rich high‐level liquid waste.

High glucose-induced p66Shc mitochondrial translocation regulates autophagy initiation and autophagosome formation in syncytiotrophoblast and extravillous trophoblast.

p66Shc, as a redox enzyme, regulates reactive oxygen species (ROS) production in mitochondria and autophagy. However, the mechanisms by which p66Shc affects autophagosome formation are not fully understood. p66Shc expression and its location in the trophoblast cells were detected in vivo and in vitro. Small hairpin RNAs or CRISPR/Cas9, RNA sequencing, and confocal laser scanning microscope were used to clarify p66Shc's role in regulating autophagic flux and STING activation. In addition, p66Shc affects mitochondrial-associated endoplasmic reticulum membranes (MAMs) formation were observed by transmission electron microscopy (TEM). Mitochondrial function was evaluated by detected cytoplastic mitochondrial DNA (mtDNA) and mitochondrial membrane potential (MMP). High glucose induces the expression and mitochondrial translocation of p66Shc, which promotes MAMs formation and stimulates PINK1-PRKN-mediated mitophagy. Moreover, mitochondrial localized p66Shc reduces MMP and triggers cytosolic mtDNA release, thus activates cGAS/STING signaling and ultimately leads to enhanced autophagy and cellular senescence. Specially, we found p66Shc is required for the interaction between STING and LC3II, as well as between STING and ATG5, thereby regulates cGAS/STING-mediated autophagy. We also identified hundreds of genes associated several biological processes including aging are co-regulated by p66Shc and ATG5, deletion either of which results in diminished cellular senescence. p66Shc is not only implicated in the initiation of autophagy by promoting MAMs formation, but also helps stabilizing active autophagic flux by activating cGAS/STING pathway in trophoblast.

Phytometabolome of Psidium guajava inhibits biofilm formation of Escherichia coli and augmented acute wound repair

Psidium guajava is an equatorial fern tree and nutritive plant with medicinal properties including antimicrobial, antioxidant, antitumor, anti-inflammatory activities, etc., In the present study, the untargeted phytometabolome of ethyl acetate and acetone extracts of P. guajava showed 107 compounds were identified using UPLC-ESI-Q-TOF-MSE analysis and assessed for anti-biofilm activity using various assays. The results clearly indicated that the ethyl acetate and acetone extracts significantly inhibit the growth and obliterate the metabolic activity of E. coli. The confocal laser scanning microscope (CSLM) analysis also confirmed the biofilm inhibitory activity of extracts against E. coli. Further, the ethyl acetate and acetone extracts displayed significant wound healing activity in BALB/c mice than the control treatment. The Doppler imaging analysis depicted that the extracts increase the blood circulation towards the wound to accelerate wound repair. Together, the UPLC-Q-TOF-MSE analysis clearly showed the metabolomics profiling of P. guajava and these metabolomes could be responsible for their potential biofilm obliterating and augmenting acute wound repair.

Characterisation of a casein-/whey protein concentrate–Antarctic krill oil emulsion system and improvement of its storage stability

Aims To develop Antarctic krill oil emulsions with casein and whey protein concentrate (WPC) and study their physicochemical properties and storage stability. Methods Emulsions were prepared by homogenisation and ultrasonication. The properties of the emulsions were investigated via ultraviolet ray spectroscopy, dynamic light scattering, confocal laser scanning microscope, sodium dodecyl sulphate–polyacrylamide gel electrophoresis, Fourier transform infra-red spectrometer, and fluorescence spectrum. Shelf life was predicted by the Arrhenius model. Results Casein- and WPC-krill oil emulsions were well formed; the mean particle diameters were less than 128.19 ± 0.64 nm and 158 ± 1.56 nm, the polymer dispersity indices were less than 0.26 ± 0.01 and 0.27 ± 0.01, and the zeta potential were around −46.88 ± 5.02 mV and −33.51 ± 2.68 mV, respectively. Shelf life was predicted to be 32.67 ± 1.55 days and 29.62 ± 0.65 days (40 °C), 27.69 ± 1.15 days and 23.58 ± 0.14 days (50 °C), 24.02 ± 0.15 days and 20.1 ± 0.08 days (60 °C). Conclusion The prepared krill oil emulsions have great potential to become a new krill oil supplement.