Confocal Laser Scanning Microscopy Images Research Articles

CASTING: A Potent Supramolecular Strategy to Cytosolically Deliver STING Agonist for Cancer Immunotherapy and SARS-CoV-2 Vaccination

CASTING: A Potent Supramolecular Strategy to Cytosolically Deliver STING Agonist for Cancer Immunotherapy and SARS-CoV-2 Vaccination

3D-Cultured Vascular-Like Networks Enable Validation of Vascular Disruption Properties of Drugs In Vitro.

Vascular-disrupting agents are an interesting class of anticancer compounds because of their combined mode of action in preventing new blood vessel formation and disruption of already existing vasculature in the immediate microenvironment of solid tumors. The validation of vascular disruption properties of these drugs in vitro is rarely addressed due to the lack of proper in vitro angiogenesis models comprising mature and long-lived vascular-like networks. We herein report an indirect coculture model of human umbilical vein endothelial cells (HUVECs) and human dermal fibroblasts (HDFs) to form three-dimensional profuse vascular-like networks. HUVECs embedded and sandwiched in the collagen scaffold were cocultured with HDFs located outside the scaffold. The indirect coculture approach with the vascular endothelial growth factor (VEGF) producing HDFs triggered the formation of progressively maturing lumenized vascular-like networks of endothelial cells within less than 7 days, which have proven to be viably maintained in culture beyond day 21. Molecular weight-dependent Texas red-dextran permeability studies indicated high vascular barrier function of the generated networks. Their longevity allowed us to study the dose-dependent response upon treatment with the three known antiangiogenic and/or vascular disrupting agents brivanib, combretastatin A4 phosphate (CA4P), and 6´-sialylgalactose (SG) via semi-quantitative brightfield and qualitative confocal laser scanning microscopic (CLSM) image analysis. Compared to the reported data on in vivo efficacy of these drugs in terms of antiangiogenic and vascular disrupting effects, we observed similar trends with our 3D model, which are not reflected in conventional in vitro angiogenesis assays. High-vascular disruption under continuous treatment of the matured vascular-like network was observed at concentrations ≥3.5 ng·ml−1 for CA4P and ≥300 nM for brivanib. In contrast, SG failed to induce any significant vascular disruption in vitro. This advanced model of a 3D vascular-like network allows for testing single and combinational antiangiogenic and vascular disrupting effects with optimized dosing and may thus bridge the gap between the in vitro and in vivo experiments in validating hits from high-throughput screening. Moreover, the physiological 3D environment mimicking in vitro assay is not only highly relevant to in vivo studies linked to cancer but also to the field of tissue regeneration.

Fabrication and stabilization mechanisms of Pickering emulsions based on gliadin/arabinoxylan complexes

In the present work, the Pickering emulsions with enhanced oxidation stability were fabricated using gliadin (G)/arabinoxylan nanoparticles (GANPs). The influence of different G/AX ratios on the properties of GANPs and corresponding physicochemical characteristics of Pickering emulsions were investigated. Results indicated that the droplet size and ζ-potential of Pickering emulsions declined with the decrease of G/AX ratios. Pickering emulsion with the smallest G/AX ratio (1:4) exhibited excellent oxidative and coalescence stability due to the formation of viscoelastic gel network, which was supported by confocal laser scanning microscopy (CLSM) images. Furthermore, the increase of salt ions in a lower concentration (0–0.2 M) was conducive to the flocculation of the droplets, while further increasing the NaCl concentration impaired the emulsion stability. Such elements revealed that G/AX complex is a promising stabilizer of Pickering emulsions with prominent antioxidant activity, which have favorable potential applications in protecting the functional properties of polyunsaturated fatty acids (PUFAs).

Green synthesis of DOX-loaded hollow MIL-100 (Fe) nanoparticles for anticancer treatment by targeting mitochondria

Fe-based metal-organic frameworks (MOFs) are promising drug delivery materials due to their large surface area, high stability, and biocompatibility. However, their drug loading capacity is constrained by their small pore size, and a further improvement in their drug capacity is needed. In this work, we report an effective and green structural modification strategy to improve drug loading capacity for Fe-based MOFs. Our strategy is to grow MIL-100 (Fe) on carboxylate-terminated polystyrene (PS-COOH) via a sustainable route, which creates a large inner cavity as well as exposure to more functional groups that benefit drug loading capacity. We employ the scanning electron microscope and transmission electron microscope to confirm the hollow structure of MIL-100 (Fe). Up to 30% of drug loading capacity has been demonstrated in our study. We also conduct cell viability tests to investigate its therapeutic effects on breast cancer cells (MDA-MB-231). Confocal laser scanning microscopy imaging confirms cellular uptake and mitochondrial targeting function of doxorubicin-loaded H-M (DOX@H-M) nanoparticles. JC-1 staining of cancer cells reveals a significant change in the mitochondrial membrane potential, indicating the mitochondrial dysfunction and apoptosis of tumor cells. Our study paves the way for the facile synthesis of hollow structural MOFs and demonstrates the potential of applying Fe-based MOFs in breast cancer treatment.

The elimination effects of lavender essential oil on Listeria monocytogenes biofilms developed at different temperatures and the induction of VBNC state.

Listeria monocytogenes is a typical foodborne pathogen that causes hard-to-treat bacterial infections, mainly due to its ability to form biofilm and enter into a viable but non-culturable state (VBNC). In this study, we investigated the removal effects of four antimicrobial agents on L. monocytogenes biofilms formed at 32°C and 10°C, analysed the resistances of the mature biofilms to antimicrobial agents, and explored the VBNC state of cells in mature biofilms induced by lavender essential oil (LEO). The results showed that the growth of L. monocytogenes was completely inhibited when 1·6% (v/v) of the LEO was added. Meanwhile, the results of the crystal violet staining and XTT reduction method indicated that different concentrations of LEO significantly reduced L. monocytogenes biofilms biomass and metabolic activities, followed by sodium hypochlorite, lactic acid, and hydrogen peroxide. Moreover, the confocal laser scanning microscopy (CLSM) images confirmed that the treated biofilms became thinner, the structure was sparse, and the appearance was blurry. More interestingly, L. monocytogenes biofilms developed at 10°C were less susceptible to the sanitizers than those formed at 32°C. In addition, LEO presented a more significant dispersing effect on the biofilm cells, and 1/2 MIC to 4 MIC of LEO could induce fewer VBNC state cells in biofilm and plankton compared with sodium hypochlorite. This study indicated that the LEO could be considered as an ideal antibiofilm agent for controlling L. monocytogenes. But we should pay attention to the resistance of the biofilms developed at low temperatures.

Evaluation of pretreatments on intra-radicular dentin bond strength of self-adhesive resin cements.

To evaluate the influence of dentin pretreatments on the push-out bond strength (POBS) of self-adhesive resin cements (SARCs) to radicular dentin. Two experimental pretreatments (2.5% titanium tetrafluoride (TiF4 ) and 26% polyacrylic acid (PA)) and two SARCs: Maxcem Elite (MAX) and Calibra Universal (CAL) were used. For each cement, a control group was applied as indicated by the manufacturer. Sixty bovine incisors were restored (n = 10) and subjected to POBS evaluation. Failure mode, adhesive interface and surface morphology were analyzed by a Scanning Electron Microscopy (SEM) and resin infiltration was performed by confocal laser scanning microscopy (CLSM). Data for POBS and CLSM were analyzed by three-way ANOVA and Bonferroni post-hoc (α = 0.05). For MAX, both experimental pretreatments resulted in increased POBS in the cervical third, and for CAL, only the PA resulted in higher POBS in the cervical third. The most failures occurred between the dentin and the resin cement, except when TiF4 was applied. For PA, analysis of surface morphology showed open dentinal tubules, while TiF4 presented particle agglomerates. SEM and CLSM images confirmed presence of resin tags for both pretreatments. Pretreating radicular dentin with PA and TiF4 solutions improves the bond strength of SARCs.

Niclosamide as a Repurposing Drug against Corynebacterium striatum Multidrug-Resistant Infections.

Corynebacterium striatum (C. striatum) is an emerging multidrug-resistant (MDR) pathogen associated with nosocomial infections. In this scenario, we screened the antimicrobial activity of the anthelmintic drugs doramectin, moxidectin, selamectin and niclosamide against 20 C. striatum MDR clinical isolates. Among these, niclosamide was the best performing drug against C. striatum. Niclosamide cytotoxicity was evaluated by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay on immortalized human keratinocyte cells (HaCaT). After 20 h of treatment, the recorded 50% cytotoxic concentration (CC50) was 2.56 μg/mL. The antibacterial efficacy was determined via disc diffusion, broth microdilution method and time-killing. Against C. striatum, niclosamide induced a growth inhibitory area of 22 mm and the minimum inhibitory concentration that inhibits 90% of bacteria (MIC90) was 0.39 μg/mL, exhibiting bactericidal action. The biofilm biomass eradicating action was investigated through crystal violet (CV), MTT and confocal laser scanning microscopy (CLSM). Niclosamide affected the biofilm viability in a dose-dependent manner and degraded biomass by 55 and 49% at 0.39 μg/mL and 0.19 μg/mL. CLSM images confirmed the biofilm biomass degradation, showing a drastic reduction in cell viability. This study could promote the drug-repurposing of the anthelmintic FDA-approved niclosamide as a therapeutic agent to counteract the C. striatum MDR infections.

Formulation and evaluation of embelin loaded nanoliposomes: Optimization, in vitro and ex vivo evaluation

The present research work designed to prepare and optimize Embelin (EMB) loaded liposomes. The liposomes were prepared by solvent evaporation method and optimized using 3-factors at 3- levels Box-Behnken design (Design-Expert software., Version 12, Stat-Ease, USA). The effect of independent variables was taken as Leciva-S70 (X1), cholesterol (X2), sonication time (X3) and their individual as well combined effects were observed on vesicle size (Y1), entrapment efficiency (Y2), and in-vitro release (Y3). The optimized formulation (EMB-NLop) was further characterized for drug release, DPPH assay, confocal laser scanning microscopy (CLSM), and ex-vivo permeation study. EMB-NLop depicted the vesicle size of 70.36 nm, PDI (0.11), entrapment efficiency of 78.22 ± 0.93% and drug release of 86.41 ± 3.34%. A better antioxidant activity of 89.27% was found as compared to standard ascorbic acid. Ex-vivo permeation studies showed significantly enhanced permeation from EMB-NLop (75.02 ± 2.05%) than EMB suspension (34.56 ± 1.73%). Further, the CLSM image of the nasal mucosa suggested rhodamine B loaded EMB-NLop showed better penetration as compared to control (rhodamine B- solution). From all experimental data, it was concluded that EMB-NLop is a promising and effective formulation for intranasal delivery.

Nanomechanical subsurface characterisation of cellulosic fibres

AbstractThe mechanical properties of single fibres are highly important in the paper production process to produce and adjust properties for the favoured fields of application. The description of mechanical properties is usually characterised via linearized assumptions and is not resolved locally or spatially in three dimensions. In tensile tests or nanoindentation experiments on cellulosic fibres, only mechanical parameter for the whole fibre, such as elastic modulus or hardness, is usually obtained. To obtain a more detailed mechanical picture of the fibre, it is crucial to determine mechanical properties in depth. To this end, we discuss an atomic force microscopy-based approach to examine stepwise the local stiffness as a function of indentation depth via static force-distance curves. To our knowledge, we are the first authors to apply this method cellulosic fibres. The method was applied to linter fibres (extracted from a finished paper sheet) as well as to natural raw cotton fibres to better understand the influence of the pulp treatment process in paper production on the mechanical properties. Both types of fibres were characterised in dry and wet conditions with respect to alterations in their mechanical properties. The used stepwise analysis method of the force-distance curves allowed subsurface imaging of the fibres. It could be revealed how the walls in the fibre structure protects the fibre against mechanical loading. Via a combined 3D display of the mapped topography and the fitted elastic moduli in z-direction, a spatially resolved mechanical map of the fibre interior near the surface could be established. Additionally, we labelled the fibres with different carbohydrate binding modules tagged with fluorescent proteins to compare the AFM results with fluorescence confocal laser scanning microscopy imaging. Nanomechanical subsurface imaging in combination with fluorescent protein labelling is thus a tool to better understand the mechanical behaviour of cellulosic fibres, which have a complex, hierarchical structure.Graphical abstract

Polyurethane-Based Porous Carbons Suitable for Medical Application.

The main aim of the study was to synthesize and analyze spectral data to determine the structure and stereometry of the carbon-based porous material internal structure. Samples of a porous biomaterial were synthesized through anionic polymerization following our own patent and then carbonized. The samples were investigated using MALDI ToF MS, FTIR ATR spectroscopy, optic microscopy, SEM, confocal laser scanning microscopy and CMT imaging. The analysis revealed the chemical and stereological structure of the obtained porous biomaterial. Then, the parameters characterizing the pore geometry and the porosity of the samples were calculated. The developed material can be used to collect adsorption of breathing phase samples to determine the parity composition of exhaled air.

Functional and molecular characterization of a cold-active lipase from Psychrobacter celer PU3 with potential antibiofilm property

The lipase gene from Psychrobacter celer PU3 was cloned into pET-28a(+) expression vector and overexpressed in E. coli BL21 (DE3) pLysS cells. The purified Psychrobacter celer lipase (PCL) was characterized as an alkaline active enzyme and has a molecular mass of around 30 kDa. The PCL was active even at a low temperature and the optimum range was observed between 10 and 40 °C temperatures. MALDI-TOF and phylogenetic analysis ensured that Psychrobacter celer PU3 lipase (PCL) was closely related to P. aureginosa lipase (PAL). MD simulation results suggest that temperature change did not affect the overall structure of PCL, but it might altered the temperature-dependent PCL functional changes. R1 (129–135 AA) and R2 (187–191 AA) regions could be important for temperature-dependent PCL function and they fluctuated much at 35 °C temperature. PMSF completely inhibited PCL lipase activity and it demonstrates the presence of serine residues in the active site of PCL. PCL is moderately halophilic and most of the tested organic solvents found to be inhibiting the lipase activity except the solvents ethanol and methanol. PCL activity was increased with surfactants (SDS and CTAB) and bleaching agents (hydrogen peroxide). The effect of different metal ions on PCL resulted that only mercuric chloride was found as the enhancer of the lipase activity. Antibiofilm property of PCL was evaluated against pathogenic Vibrio parahaemolyticus isolated from the diseased shrimp and MIC value was 500 U. PCL significantly altered the morphology and biofilm density of V. parahaemolyticus and the same was observed through scanning electron microscope (SEM) and confocal laser scanning microscope (CLSM) imaging. RT-PCR analysis revealed that the mRNA expression level of biofilm, colony morphology and major toxin-related (aphA, luxS, opaR, tolC, toxR) genes of V. parahaemolyticus were significantly downregulated with PCL treatment.

Combined Anti-Biofilm Enzymes Strengthen the Eradicate Effect of Vibrio parahaemolyticus Biofilm: Mechanism on cpsA-J Expression and Application on Different Carriers.

Vibrio parahaemolyticus is a human foodborne pathogen, and it can form a mature biofilm on food and food contact surfaces to enhance their resistance to antibacterial agents. In this study, the effect of anti-biofilm enzymes (combined lipase, cellulase and proteinase K) on the inhibition and eradication of pathogen biofilm was evaluated. The biofilm content of V. parahaemolyticus showed the highest level at the incubation time of 24 h, and the combined enzymes significantly inhibited the biofilm’s development. The biofilm’s inhibition and eradication rate at an incubation time of 24 h was 89.7% and 66.9%, respectively. The confocal laser scanning microscopic images confirmed that the microcolonies’ aggregation and the adhesion of biofilm were inhibited with the combined enzyme treatment. Furthermore, combined enzymes also decreased the concentration of exopolysaccharide (EPS) and disrupted the EPS matrix network, wherein the expression of the EPS-related gene, cpsA-J, was likewise suppressed. The combined enzymes showed an excellent inhibition effect of V. parahaemolyticus biofilm on different carriers, with the highest inhibition rate of 59.35% on nonrust steel plate. This study demonstrates that the combined enzyme of lipase, cellulase and proteinase K could be a novel candidate to overcome biofilm’s problem of foodborne pathogens in the food industry.

Efficacy of Salvadora persica root extract as an endodontic irrigant – An in-vitro evaluation

IntroductionThe main reason for endodontic failure is bacteria and chemical irrigation using chemicals in the form of irrigation has been deemed effective. The objectives of this study were to evaluate the in vitro antimicrobial effects of Salvadora persica extract as an endodontic irrigant and to compare it with sodium hypochlorite (NaOCl). MethodsIn this study, stock solutions of 10 mg/ml of ethanolic Salvadora persica extract and 1% of sodium hypochlorite were serially diluted three times and then exposed to alamarBlue® cell viability testing on human gingival fibroblast cells. Antimicrobial investigations used Crystal violet assay and live/dead® Baclight™ assay on Pseudomonas aeruginosa and E. faecalis biofilms followed by confocal laser scanning microscopy (CLSM) imaging. The results were further analyzed using appropriate statistical tests. Values of p ≤ 0.05 were considered statistically significant. ResultsThe Salvadora persica root extract was cytotoxic to gingival fibroblasts but significantly weaker than NaOCl at its highest concentration. Antimicrobial activity was observed for both irrigation solutions, but NaOCl has significantly stronger antimicrobial effects in the live/dead® assay. NaOCl could reduce the bacterial viability for both species more than Salvadora persica extract in the Crystal violet assay. There were limited effects of Salvadora persica extract on Pseudomonas aeruginosa; these effects were confirmed on CLSM images. ConclusionEthanolic Salvadora persica root extract demonstrated cytotoxic and antimicrobial effects that were significantly weaker than NaOCl exposure. These variations in effects might introduce the herbal extract as an alternative irrigant especially for regenerative endodontic treatments that require great care for cellular components.

Preparation of Natural Food-Grade Core-Shell Starch/Zein Microparticles by Antisolvent Exchange and Transglutaminase Crosslinking for Reduced Digestion of Starch.

The purpose of this study was to slow down the digestibility of starch granules by encapsulating it in zein shells. Drop of the preformed swollen corn starch (CS) granule suspension into thermal-treated zein ethanolic solution enables antisolvent precipitation of thermal-treated zein on the surface of the preformed swollen CS granules, leading to the formation of core-shell starch/zein microparticles. Confocal laser scanning microscopy images showed that the preformed swollen CS granules were coated by thermal-treated zein shells with a thickness of 0.48–0.95 μm. The volume average particle diameter of core-shell starch/zein microparticles was 14.70 μm and reached 18.59–30.98 μm after crosslinking by transglutaminase. The results of X-ray diffraction and Fourier transform infrared spectroscopy demonstrated that an interaction occurred between the preformed swollen CS granules and the thermal-treated zein. The results for thermodynamic characteristics, pasting properties, and swelling power indicated that the compact network structure of core-shell starch/zein microparticles crosslinked by transglutaminase could improve starch granule thermal stability and resistance to shearing forces. Compared to native CS, the peak gelatinization temperatures of core-shell starch/zein microparticles increased significantly (p < 0.05), with a maximum value of 76.64°C. The breakdown values and the swelling power at 95°C of core-shell starch/zein microparticles significantly (p < 0.05) decreased by 52.83–85.66% and 0.11–0.28%, respectively. The in vitro digestibility test showed that the contents of slowly digestible starch and resistant starch in the core-shell starch/zein microparticles increased to ∼42.66 and ∼34.75%, respectively, compared to those of native CS (9.56 and 2.48%, respectively). Our research supports the application of food-grade core-shell starch/zein microparticles to formulate low-digestibility food products.

Stabilisation of water in water Pickering emulsion containing gelatin and maltodextrin by bitter vetch protein nanoparticles

SummaryThe incompatibility between water soluble polymers is one of the interactions between the different constituents of the food systems, which eventually leads to system instability due to phase separation. The use of natural nanoparticles under the Pickering mechanism is a promising technique for the stabilisation of water‐in‐water (W/W) emulsions. In this study, bitter vetch protein nanoparticles (BPNs), which was produced by ultrasonic treatment, was used to stabilise W/W emulsion. Then additional tests were performed to investigate the structure and properties of the nanoparticles and stabilised emulsion. Scanning electron microscopy (SEM) images showed the size of BPNs to be about 60 nm. The zeta potential of protein nanoparticle solution was about 50 mV. Fourier transform infrared spectroscopy (FTIR), X‐ray diffraction (XRD) and simultaneous thermal analysis (STA) confirmed that chemical structure and thermal behaviour of BPNs did not change during nanoparticle preparation process. BPNs could stabilise W/W emulsion containing gelatin and maltodextrin. Confocal laser scanning microscopy (CLSM) images confirmed the stability of gelatin/maltodextrin emulsion containing BPNs. The drop test confirmed the presence of maltodextrin in the continuous phase of the emulsion. The stabilised W/W emulsion had 100% stability against stress and temperature of 80 °C. Rheology analysis showed the viscoelastic nature and pseudoplastic behaviour of the stabilised emulsion.

Toxic effects of microplastics in plants depend more by their surface functional groups than just accumulation contents

Differentially charged microplastics (MPs) engendered by plastic aging (e.g., plastic film) widely existed in the agricultural ecosystem, yet minimal was known about the toxic effects of MPs on plants and their absorption and accumulation characteristics. Root absorption largely determined the migration and accumulation risks of MPs in the soil-crop food chain. Here, five types of MPs exposure experiments of leaf lettuce were implemented to simulate root absorption by hydroponics. MPs exposure caused different degrees of growth inhibition, root lignification, root cell apoptosis, and oxidative stress responses; accelerated chlorophyll decomposition and hampered normal electron transfer within the PSII photosystem. Moreover, the uptake of essential elements by roots was inhibited to varying degrees due to the pore blockage in the cell wall and the hetero-aggregation of opposite charges after MPs exposure. MPs exposure observably up-regulated the organic metabolic pathways in roots, thus affecting MPs mobility and absorption through the electrostatic and hydrophobic interactions between the root exudations and MPs. Importantly, MPs penetrated the root extracellular cortex into the stele and were transported to the shoots by transpiration through xylem vessels based on confocal laser scanning microscopy and scanning electron microscopy images. Quantitative analysis of MPs indicated that their toxic effects on plants were determined to a greater extent by the types of surface functional groups than just their accumulation contents, that is, MPs were confirmed edible risks through crop food chain transfer, but bioaccumulation varied by surface functional groups.

Chemical composition, antimicrobial, and antioxidant cytotoxic activities of essential oil from Actinidia arguta

Chemical composition, antimicrobial, antioxidant, and cytotoxic properties of Actinidia arguta essential oil (AEO) were evaluated. Gas chromatography-mass spectrometry analysis identified 56 chemical compounds, with the most abundant being Squalene (23.08%), γ-sitrostorol (8.10%), and β-Tocopherol (7.01%). Whereas the AEO had significant antimicrobial activity against Staphylococcus aureus and Saccharomyces cerevisiae, it showed mild efficacy against Bacillus subtilis and Microsporum canis. On the contrary, the Gram-negative bacteria, Escherichia coli and Pseudomonas aeruginosa, were not susceptible to the AEO pressure. On the other hand, the AEO exhibited strong antioxidant activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH), β-carotene, and hydroxyl radicals, with IC50 values of 117.60, 73.60, and 35.15μg/mL, respectively. Additionally, compared to the PC-3 or HT-29 cell lines, the A549 cells were more susceptible to the AEO (IC50; 6.067mg/mL). Besides, the confocal laser scanning microscopy imaging showed that 16mg/mL of the AEO-induced apoptosis in the A549 cell lines. Our data indicated that the AEO might be useful in the food and pharmaceutical industry. Preparation of Actinidia arguta essential oil (a) and schematic overview of the experiment (b).

Low-sodium salt mediated aggregation behavior of gluten in wheat dough.

Reducing sodium in foods has attracted the attention of consumers, it is therefore necessary to explore sodium alternatives (i.e., low-sodium salt). However, the mechanism of low-sodium salt on gluten in dough remains unclear. Effect of low-sodium salt on the aggregation behaviors of gluten in dough was investigated and compared with those with NaCl and KCl in this study. The results showed that low-sodium salt enhanced gluten strength and prolonged gluten aggregation time. Low-sodium salt decreased the content of SDS extractable protein under non-reducing conditions. Low-sodium salt changed the spatial conformation of gluten by reducing β-turn structure and increasing β-sheet structure. Confocal laser scanning microscopy images indicated that low-sodium salt promoted the formation of a larger and dense gluten network. In summary, this study showed that low-sodium salt promoted the aggregation of gluten in dough, and the change of gluten structure explained this aggregation mechanism. Its mode of action was similar to NaCl and KCl, which provided a theoretical basis for the study of sodium substitutes in flour products.

High-humidity hot air impingement blanching (HHAIB): An emerging technology for tomato peeling

Peeling is an essential operation for tomato processing. A new peeling method, high-humidity hot air impingement blanching (HHAIB) heating technology, was developed as an alternative to the conventional lye and hot-water peeling to eliminate the use of chemicals and the discharge of wastewater. The current work explored the feasibility of HHAIB for tomato peeling. The effects of heating temperature (100–120 °C), relative humidity (20%–40%) and heating time (0–180 s) on the peeling performance were investigated. The optimum treatment was found to be 110 °C heating temperature in combination with 40% of relative humidity and 75 s treatment time, which resulted in lower peeling loss, firmness loss and color deterioration compared with other HHAIB conditions that achieved 100% peelability. The comparative study of optimized HHAIB peeling with conventional lye and hot-water peeling showed that HHAIB peeled tomato obtained lower peeling loss and firmness loss, and higher preservation of phytochemicals, antioxidant capacity and color. In addition, compared with fresh tomatoes, HHAIB processing increased the antioxidant activity, lycopene, and total phenolic content in peeled tomatoes by 16.01%, 10.46%, 12.80%, respectively. The laser scanning confocal microscopy image of fresh tomato skin surface and the scanning electron microscope images of peels and flesh showed that HHAIB caused cracking of the epidermis and melting of the cuticular membrane while reduced the serious damage of flesh. Industrial practicePeeling is a necessary step in tomato processing, which impacts subsequent processing efficiency and product quality. At present, the most common used peeling methods in the industry are hot water or/and alkali peeling, but it induces the loss of water-soluble nutrients, chemical residues and waste liquid treatment. Therefore, the industry urgently needs an alternative peeling technology. The current work shows that HHAIB is a very promising peeling technology as it not only has an excellent peeling performance, but also enhances the preservation of phytochemicals, antioxidant capacity and quality attributes compared to conventional lye and hot-water peeling.

Controlling Listeria monocytogenes Growth and Biofilm Formation Using Flavonoids

The aim of this study was to investigate the ability of natural plant-derivate products (flavonoid compounds) to inhibit the growth and biofilm-forming ability of Listeria monocytogenes. A collection of 500 synthetic and natural flavonoids were tested individually on strains of L. monocytogenes for their antimicrobial and antibiofilm activity. The flavonoids were tested against a L. monocytogenes cocktail of five strains at a concentration of 100 μM to determine their effect on planktonic growth. The optical density was measured every hour for 24 h at 37°C, and every hour for 48 h at 22°C. A total of 17 flavonoids were chosen for further study because of their ability to significantly reduce the growth of L. monocytogenes up to 97%. An additional two flavonoids that increased planktonic growth were chosen as well to investigate whether they had the same effect on biofilm growth. A lower concentration of flavonoid compounds (50 μM) was selected to investigate the individual effects on L. monocytogenes biofilm formation using (i) stainless steel coupons to quantify biomass using crystal violet staining and (ii) glass slides using confocal laser scanning microscopic (CLSM) imaging to observe the biofilm architecture. The 19 flavonoids showed various levels of L. monocytogenes biofilm growth inhibition, ranging from 2 to 100% after 48 h of incubation at 22 or 10°C. This includes 18 of the 19 flavonoids significantly (P ≤ 0.05) inhibiting L. monocytogenes biofilm formation on stainless steel coupons under at least one of the testing conditions. However, only one flavonoid compound demonstrated significant biofilm inhibition (P ≤ 0.05) under all conditions tested. Furthermore, 8 of the selected 19 flavonoid compounds showed visible reductions through CLSM in L. monocytogenes biofilm formation. Overall, we identified five flavonoid compounds to be promising antibiofilm and antimicrobial agents against L. monocytogenes.