Search Results (351)

Biofilms cause a variety of problems, such as food spoilage, food poisoning, infection, tooth decay, periodontal disease, and metal corrosion, so knowledge on biofilm prevention and removal is important. A detailed observation of the three-dimensional structure of biofilms on the nanoscale is expected to provide insight into this. In this study, we report on the successful in situ nanoscale observations of a marine bacterial biofilm on glass in phosphate buffer solution (PBS) using both scanning ion conductance microscopy (SICM) and confocal laser scanning microscopy (CLSM) over the same area. By observing the same area by SICM and CLSM, we were able to clarify the three-dimensional morphology of the biofilm, the arrangement of bacteria within the biofilm, and the difference in local ion conductivity within the biofilm simultaneously, which could not be achieved by observation using a microscope alone. Full article

Although corn germ meal is a rich source of dietary fiber, it contains a relatively low proportion of soluble dietary fiber (SDF) and is frequently contaminated with high levels of zearalenone (ZEN). Solid-state fermentation has the dual effects of modifying dietary fiber (DF) and degrading mycotoxins. This study optimized the solid-state fermentation process of corn germ meal using Bacillus subtilis K6 through response surface methodology (RSM) to enhance SDF yield while efficiently degrading ZEN. Results indicated that fermentation solid-to-liquid ratio and time had greater impacts on SDF yield and ZEN degradation rate than fermentation temperature. The optimal conditions were determined as temperature 36.5 °C, time 65 h, and solid-to-liquid ratio 1:0.82 (w/v). Under these conditions, the ZEN degradation rate reached 96.27 ± 0.53%, while the SDF yield increased from 9.47 ± 0.68% to 20.11 ± 1.87% (optimizing the SDF/DF ratio from 1:7 to 1:3). Scanning electron microscopy (SEM) and confocal laser scanning microscope (CLSM) revealed the structural transformation of dietary fiber from smooth to loose and porous forms. This structural modification resulted in a significant improvement in the physicochemical properties of dietary fiber, with water-holding capacity (WHC), oil-holding capacity (OHC), and water-swelling capacity (WSC) increasing by 34.8%, 16.4%, and 15.2%, respectively. Additionally, the protein and total phenolic contents increased by 23.0% and 82.61%, respectively. This research has achieved efficient detoxification and dietary fiber modification of corn germ meal, significantly enhancing the resource utilization rate of corn by-products and providing technical and theoretical support for industrial production applications. Full article

The continuous casting of Ti-Nb microalloyed steel was simulated with high temperature confocal laser scanning microscopy (HTCLSM). Evolution of the sample surface morphology was observed in-situ, during cooling conditions chosen to represent different locations in a cast slab. Calculations with a thermodynamics model of carbonitride precipitate formation agreed with the transmission electron microscopy (TEM) analysis that fine reliefs observed on the sample surface were actually caused by interior precipitation of (Ti,Nb)(C,N). Precipitation and the resulting reliefs changed with location beneath the slab surface, simulated casting speed, and steel composition. With the same casting speed and steel composition, reliefs in the simulated slab surface sample appeared earlier and were larger than in the slab center. With increased casting speed, reliefs were observed later and decreased in size. With increased titanium or niobium content, reliefs appeared earlier and increased in number. TEM measurement showed that the precipitate diameters were mainly smaller than 4 nm, with a few between 4 and 8 nm. The property of surface reliefs observed via HTCLSM correlated qualitatively with the number and size of internal precipitates measured with TEM, showing this to be an effective tool for indirectly characterizing nanoscale secondary phase precipitation inside the sample. Full article

Fresh apple pulp from the Granny Smith variety was used at different levels (5–15% w/w) for yogurt production. Color, texture, microstructure, aroma, and sensory analyses were used to evaluate the effect of the apple pulp on the main characteristics of yogurt. Yogurts with apple pulp presented a lower brightness (L*) and an increased redness (a*) and yellowness (b*), which were significantly affected by the apple pulp concentration. The texture analysis revealed an improved consistency and reduced syneresis, leading to a creamier and more stable product. The aroma profile of yogurts was enriched, presenting higher ester contents. Confocal laser scanning microscopy showed that the incorporation of modest quantities of apple pulp resulted in the formation of initially denser networks, while at elevated levels, an enhanced microscopic phase separation occurred. A 5% apple pulp addition achieved a balance between enhancing flavor and texture retention while maintaining high overall acceptability, as was also confirmed by the sensory evaluation. Full article

Cardiotoxicity related to anthracyclines and trastuzumab represents a significant clinical challenge in cancer therapy, often limiting treatment efficacy and patient survival. The underlying mechanisms of cardiotoxicity involve the activation of NLRP3 and the MyD88-dependent signaling pathway. Proprotein convertase subtilisin/kexin type 9 inhibitors (PCSK9i), such as inclisiran, are known for their lipid-lowering effects, but emerging data indicate that they may also exert pleiotropic benefits beyond cholesterol reduction. This study investigates whether inclisiran can mitigate the cardiotoxic effects of anthracyclines and trastuzumab through reduction of NLRP3 activation and MyD88 signaling, independently of its effects on dyslipidemia. Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) were exposed to subclinical concentrations of doxorubicin (1 µM) and trastuzumab in sequential therapy (200 nM), alone or in combination with inclisiran (100 nM) for 24 h. After the incubation period, we performed the following tests: determination of cardiomyocytes apoptosis, analysis of intracellular reactive oxygen species, lipid peroxidation products (including malondialdehyde and 4-hydroxynonenal), intracellular mitofusin-2 and Ca⁺⁺ levels. Troponin and BNP were quantified through selective ELISA methods. A confocal laser scanning microscope was used to study cardiomyocyte morphology and F-actin staining after treatments. Moreover, pro-inflammatory studies were also performed, including the intracellular expression of NLRP-3, MyD-88 and twelve cytokines/growth factors involved in cardiotoxicity (IL-1α, IL-1β, IL-2, IL-4, IL-6, IL-10, IL-12, IL17-α, IFN-γ, TNF-α, G-CSF, GM-CSF). Inclisiran co-incubated with doxorubicin and trastuzumab exerts significant cardioprotective effects, enhancing cell viability by 88.9% compared to only DOXO/TRA treated cells (p < 0.001 for all). Significant reduction of oxidative stress, and intracellular levels of NLRP-3, MyD88, IL-1α, IL-1β, IL-6, IL-12, IL17-α, TNF-α, G-CSF were seen in the inclisiran group vs. only DOXO/TRA (p < 0.001). For the first time, PCSK9i inclisiran has been shown to exert significant anti-inflammatory effects to reduce anthracycline-HER-2 blocking agent-mediated cardiotoxicity through NLRP-3 and Myd-88 related pathways. The overall conclusions of the study warrant further investigation of the use of PCSK9i in primary prevention of CTRCD in cancer patients, independently from dyslipidemia. Full article

This study is based on the laser-induced thermal-crack propagation (LITP) technology, focusing on the issues of deviation and thermal damage during the transverse crack propagation process, with the aim of achieving high-purity, non-destructive, and high-precision cutting of glass. A 50 W, 1064 nm fiber laser is used for S-pattern scanning cutting of soda–lime glass. A moving heat source model is established and analyzed via MATLAB R2022a numerical simulation. Combined with the ABAQUS 2019 software, the relationships among temperature field, stress field, crack propagation, and deviation during laser-induced thermal crack cutting are deeply explored. Meanwhile, laser thermal fracture experiments are also carried out. A confocal microscope detects glass surface morphology, cross-sectional roughness and hardness under different heat flux densities (HFLs), determining the heat flux density threshold affecting the glass surface quality. Through a comprehensive study of theory, simulation, and experiments, it is found that with an increase in the HFL value of the material, the laser-induced thermal crack propagation can be divided into four stages. When the heat flux density value is in the range of 47.2 to 472 W/m², the glass substrate has good cross-sectional characteristics. There is no ablation phenomenon, and the surface roughness of the cross-section is lower than 0.15 mm. The hardness decreases by 9.19% compared with the reference value. Full article

The peening process plays a pivotal role in enhancing the properties of aluminium alloys across various industries, including aerospace, automotive, and construction. Among the critical factors influencing this process, the shot peening time is of paramount importance for studying material characteristics. In the present study, we undertook a comprehensive investigation into the mechanical properties, surface roughness, and damage evolution behaviour of 7075 aluminium alloy subjected to different shot peening durations. This investigation was conducted using a microhardness tester, laser confocal microscope, scanning electron microscope, and other advanced equipment, in conjunction with digital image correlation methods and temperature evolution analysis. Our findings demonstrate that the shot peening time has a profound impact on the mechanical properties of the 7075 alloy. Specifically, the microhardness, tensile strength, and surface roughness of the alloy increased with increasing shot peening time, whereas the elongation rate exhibited a non-monotonic trend, initially decreasing and then increasing. Utilising DIC and temperature evolution analysis, we analysed the influence of shot peening time on the damage evolution behaviour of the alloy and developed tensile damage evolution equations tailored to different shot peening durations. The damage evolution of the 7075 alloy under various shot peening times was observed to proceed through two distinct stages: smooth development and rapid damage. Notably, the damage evolution laws derived from both techniques exhibited good consistency and agreement. The present study serves as a theoretical foundation for exploring the surface peening and damage evolution of 7075 aluminium alloy, which holds significant implications for optimising peening parameters and predicting material life in engineering applications. Full article

Background: Cancer vaccines represent a groundbreaking advancement in cancer immunotherapy, utilizing tumor antigens to induce tumor-specific immune responses. However, challenges like tumor-induced immune resistance and technical barriers limit the widespread application of predefined antigen vaccines. Here, we investigated the potential of viral protein R (Vpr) peptides as effective candidates for constructing anonymous antigen vaccines in situ by directly injecting at the tumor site and releasing whole-tumor antigens, inducing robust anti-tumor immune responses to overcome the limitations of predefined antigen vaccines. Methods: The cytotoxic effects of Vpr peptides were evaluated using the CCK8 reagent kit. Membrane penetration ability of Vpr peptides was observed using a confocal laser scanning microscope and quantitatively analyzed using flow cytometry. EGFR levels in the cell culture supernatants of cells treated with Vpr peptides were evaluated using an ELISA. Surface exposure of CRT on the tumor cell surface was observed using a confocal laser scanning microscope and quantitatively analyzed using flow cytometry. The secretion levels of ATP from tumor cells were evaluated using an ATP assay kit. HMGB1 release was evaluated using an ELISA. Mouse (Male C57BL/6 mice aged 4 weeks) MC38 and LLC bilateral subcutaneous tumor models were established to evaluate the therapeutic effects of Vpr peptides through in situ vaccination. Proteomic analysis was performed to explore the mechanism of anti-tumor activity of Vpr peptides. Results: Four Vpr peptides were designed and synthesized, with P1 and P4 exhibiting cytotoxic effects on tumor cells, inducing apoptosis and immunogenic cell death. In mouse tumor models, in situ vaccination with Vpr peptide significantly inhibited tumor growth and activated various immune cells. High-dose P1 monotherapy demonstrated potent anti-tumor effects, activating DCs, T cells, and macrophages. Combining ISV of P1 with a CD47 inhibitor SIRPαFc fusion protein showed potent distant tumor suppression effects. Proteomic analysis suggested that Vpr peptides exerted anti-tumor effects by disrupting tumor cell morphology, movement, and adhesion, and promoting immune cell infiltration. Conclusions: The designed Vpr peptides show promise as candidates for in situ vaccination, with significant anti-tumor effects, immune activation, and favorable safety profiles observed in mouse models. In situ vaccination with Vpr-derived peptides represents a potential approach for cancer immunotherapy. Full article

Microbially induced concrete deterioration (MID) poses a significant and urgent challenge to urban sewerage systems globally, particularly in tropical coastal regions. Despite the acknowledged importance of biofilms in MICC, limited research on sewer pipe biofilms has hindered a comprehensive understanding of their deterioration mechanisms. To overcome this limitation, our research employed multiple staining techniques and digital volume correlation (DVC) technology, creating a new method to analyze the microstructure of biofilms, precisely identify the components of EPSs, and quantitatively examine MID mechanisms from a microscopic viewpoint. Our results revealed that the biofilm on concrete surfaces regulates the types of amino acids, thereby creating an environment conducive to microbial aggregate survival. Additionally, salinity significantly influences biofilm component distribution, while proteins play a pivotal role in biofilm mechanical stability. Notably, a high salinity fosters microbial migration within the biofilm, exacerbating deterioration. Through this multidimensional inquiry, our study established an advanced echelon of comprehension concerning the intricate mechanisms underpinning MICC. Meanwhile, by peering into the biofilms and elucidating their interplay with concrete, our findings offer profound insights, which can aid in devising strategies to counter urban sewer system deterioration. Full article

With the rapid development of the livestock farming industry, the treatment of livestock farming wastewater has become increasingly important. The microbial-algal biofilm method has gained widespread attention for cattle wastewater treatment owing to its non-toxic nature, resistance to shock loading, and high treatment efficiency. In this study, three types of substrates—polyurethane sponge, ceramic material, and moving bed biofilm reactor media—were evaluated. The formation of biofilms was assessed through variations in chlorophyll content, microscopic observations, and measurements of biofilm dry weight and attachment rate. Biofilm characterization on the different substrates was conducted via Fourier transform infrared spectroscopy, confocal laser scanning microscopy, and scanning electron microscopy. The results demonstrated that polyurethane sponge was the most effective substrate. Furthermore, a single-factor experiment was conducted to optimize the cultivation conditions for the microbial-algal biofilms and identify the optimal parameters based on the ability of the biofilm to remove COD, TN, TP, and NH₄⁺-N. The optimal conditions were as follows: an illumination intensity of 8000 lux, red light, a temperature of 20 °C, a pH of 7, and an aeration intensity of 8 L/min. Under these conditions, the pollutant removal rates were exceptionally high: ~73.4% for COD, 51.8% for TP, 57.0% for TN, and 75.1% for NH₄⁺-N. Full article

The martensitic transformation mechanism in the heat-affected zone of DP1180 steel plays a decisive role in the strength of welded joints. In this work, the nucleation and growth kinetics of martensite laths in the coarse grain heat-affected zone (CGHAZ) are analyzed by a high-temperature laser scanning confocal microscope (LSCM). The grain distribution and stress distribution of the samples after in situ observation are analyzed by electron backscatter diffraction (EBSD). The results reveal that austenite grain growth is realized by continuous grain boundary annexation and grain boundary migration of small grains by large grains during the heating process. Seven growth modes of CGHAZ martensitic laths under laser welding conditions are proposed. Additionally, the end growth of martensitic laths is mostly attributed to the collision with grain boundaries or other laths to form a complex interlocking structure. The results of this study could provide important data support for the development of dual-phase steel materials and improvement of welding quality. Full article

Drought and metal pollution severely impact plant growth. Root-associated extremophilic fungi can improve plant performance, and their encapsulation improves protection and effectiveness. This study optimized the encapsulation conditions for an extremophilic fungus with plant growth-promoting traits using alginate–chitosan capsules. An endophytic fungus was isolated from the roots of Neltuma chilensis from the Atacama Desert and identified via internal transcribed spacer (ITS) sequencing. Its plant growth-promoting traits, including exopolysaccharide, ammonium, siderophore, and indole acetic acid production and phosphorus solubilization, were evaluated. Freeze-dried Penicillium nalgiovense was encapsulated using jet-breaking extrusion, and capsule morphology and fungal survival were assessed via scanning electron microscope (SEM), confocal laser scanning microscopy (CLSM), and viability tests. Using Taguchi’s design, optimal conditions for sphericity (0.914 ± 0.002) and mean size (3.232 ± 0.087 mm) were achieved with 1% chitosan, a 5 cm distance to the gelation bath, and a 40 Hz vibration frequency. CLSM analysis confirmed the presence of the chitosan outer layer, revealing the capsule’s coating material encapsulating the fungus P. nalgiovense. The encapsulated fungus remained viable across disinfection times, demonstrating effective protection and gradual release. These findings emphasize the need for precise parameter control in fungal encapsulation, providing a basis for developing robust bioinoculants to support plant resilience in extreme environments. Full article

The microstructures of microalgae biofilms affect biofilms’ growth, which is critical in developing efficient microalgae biofilm-based culture systems. Herein, the microstructures of Tetradesmus obliquus biofilms cultured under different nutrients were in situ observed with confocal laser scanning microscopes. The surface structures and internal pore structures for these biofilms were determined quantitatively. The results indicated that the surfaces of these biofilms were all wrinkled with many folds in micrometres. The biofilms cultured under BG11, BG11^+glycerine, BG11^+urea, and BG11^+NH₄Cl nutrients had small pores with diameters of 15~30 μm, whereas the biofilms formed under BG11^+NaHCO₃ and BG11^+NaNO₃ presented many large pores with diameters of 50~150 μm. The mechanism of forming different microstructures of these nutrients was interpreted by analyzing cell surface properties. We found that the cells cultured under BG11^+NaHCO₃ and BG11^+NaNO₃ had more hydrophobic surface groups. The high cell–cell interactions between hydrophobic cells made cells tend to aggregate together and form biofilms with more inner pores, which may be conducive to cell growth and biofilm development. The study offers new insights into understanding the microstructural characteristics of microalgae biofilms cultured under different nutrients, providing important guidance for the development of biofilm-based culture systems. Full article

As the crucial core material in aluminum–plastic-laminated films, aluminum foil serves as a barrier and shaping element for lithium-ion battery soft packaging. However, its thinness, measuring only tens of microns, makes it susceptible to the formation of pinholes during the manufacturing process, which can significantly impact the barrier performance and properties of the aluminum–plastic-laminated film. The morphology and composition of foreign particles that lead to pinholes were analyzed using scanning electron microscopy (SEM) with energy-dispersive spectroscopy (EDS). Additionally, the formation mechanism and evolution law of pinholes were investigated using a laser scanning confocal microscope (LSCM). The results revealed that foreign particles responsible for pinholes originated from the inclusions in the aluminum alloy melt, filter aid particles from rolling oil, and environmental dust particles. To address this issue, potential strategies for controlling foreign particles were proposed. These included purifying the aluminum alloy melt, filtering the rolling oil, and maintaining a clean production environment. The simulated experiments showed that foreign particles were gradually embedded in the aluminum matrix during plastic deformation, leading to damage in the aluminum matrix. When the cumulative rolling reduction ratio exceeded 38%, the aluminum foil and foreign particles began to separate along the rolling direction, resulting in the formation of pinholes. The mechanism of uncoordinated deformation between foreign particles and aluminum foil was elaborated in detail. In addition, the simulation experiment indicated that once the cumulative reduction ratio surpassed 50%, the aspect ratio of the pinhole increased rapidly. When the cumulative reduction ratio increased to 83%, the pinhole began to gradually heal. Consequently, a quantitative relationship model between the pinhole area and the rolling reduction ratio was constructed. The pinhole evolution model enables a rough prediction of the actual pinhole area change and meets the requirements for engineering applications. This research provides both engineering applications and theoretical prediction approaches that can aid in the production of high-quality aluminum foil for lithium-ion battery soft packaging. Full article

This paper aims to present the phenotypic characteristics, such as length, width, circular diameter, volume, surface area to cross-sectional area ratio, surface uniformity, and surface texture, of the foodborne pathogens Salmonella enterica serovar Agona, Salmonella enterica serovar Enteritidis, Listeria monocytogenes, and Campylobacter jejuni. It is a novel “field-guide” presentation of the observable morphological characteristics of these four species in the form of a Dichotomous Key. Phenotypic values of the pathogens were measured using a high-accuracy, nano-level-resolution 3D laser confocal scanning microscope. All samples were fixed with glutaraldehyde and stained with methylene blue, ensuring morphological preservation. The images of the pathogens were captured under various imaging modes, including 3D, laser, and transmission electron microscopy resolutions. Results show that these pathogens exhibit distinct morphological and surface properties, with Campylobacter jejuni showing unique features. The dichotomous key shows a simplified way of classifying selected foodborne pathogens from their morphometric properties. The morphometric measurements, in many instances first reported in the literature, and images provide a readily observable way to identify and classify microorganisms, allowing researchers to potentially study evolutionary relationships, assess species diversity, and understand how organisms interact with their environment, especially when genetic information is limited or difficult to obtain. Full article