Confocal Microscopy Observations Research Articles

Encapsulation of zingerone by self-assembling peptides derived from fish viscera: Characterization, interaction and effects on colon epithelial cells

The purpose of the present work was to encapsulate zingerone (a bioactive compound from ginger) by self-assembling peptides derived from fish viscera. The encapsulation conditions were investigated and the structure of fish peptides-zingerone complex was characterized. The interaction between zingerone and fish peptides was investigated using fluorescence spectroscopy. Further research was performed on the in vitro release of zingerone and fish peptide-zingerone as well as their antiproliferative effects on colon epithelial Caco-2 cells. The results demonstrated that zingerone can be successfully encapsulated by self-assembling peptides derived from fish viscera with high encapsulation efficiency and loading capacity. Furthermore, transmission electron microscope and confocal laser scanning microscope observations revealed the successful encapsulation of zingerone by fish viscera peptides. In addition, in vitro release and antiproliferative activity against Caco-2 cells can be significantly increased by encapsulating zingerone via peptide self-assembly. The current study advances knowledge of encapsulation of bioactive compounds through peptide self-assembly.

Investigation on the Preparation and Performances of Epoxy-Modified Asphalt Binder and Its Mixtures.

Epoxy-modified asphalt binder has been widely used in steel deck pavement due to its excellent properties and it is a potential candidate for long life pavements. However, its short reserve time limits its widespread application in pavement engineering. Therefore, this work developed a novel epoxy-modified asphalt binder composed of a laboratory-made curing agent as a solution. Firstly, optimization of preparation temperature of this new material was studied to balance the requirements of enough construction time and the material strength and elongation. The epoxy-modified asphalt binder, prepared at the optimal temperature of 140 °C, had a reserve time exceeding 120 min, whereas the tensile strength and the elongation at failure were 2.22 MPa and 216%, respectively, which satisfied the standard requirements of paving epoxy material well. Secondly, the asphalt mixture property tests demonstrate excellent high-temperature rutting resistance, water stability and low-temperature anti-cracking ability. Additionally, the compatibility and colloidal stability of this epoxy-modified asphalt binder were analyzed in terms of microphase structure. The uniform microphase distribution of this binder showed by the laser confocal microscope observation in both short-term aging case and long-term aging case, indicates the great compatibility between asphalt and epoxy resin during paving process and service life. Furthermore, fatigue tests were conducted to evaluate the long-term durability. The fatigue life of epoxy-modified asphalt mixtures increased by 435%, 427%, 342%, and 276% under the stress ratios of 0.3, 0.4, 0.5, and 0.6, respectively, compared to those of SBS-modified asphalt mixtures. All these results indicate that the new epoxy-modified asphalt material is promising for applications in pavement engineering, especially suitable for long-life road pavement.

Micro tensile tester measurement of biomechanical properties and adhesion force of microtubule-polymerization-inhibited cancer cells

Both mechanical and adhesion properties of cancer cells are complex and reciprocally related to migration, invasion, and metastasis with large cell deformation. Therefore, we evaluated these properties for human cervical cancer cells (HeLa) simultaneously using our previously developed micro tensile tester system. For efficient evaluation, we developed image analysis software to modify the system. The software can analyze the tensile force in real time. The modified system can evaluate the tensile stiffness of cells to which a large deformation is applied, also evaluate the adhesion strength of cancer cells that adhered to a culture substrate and were cultured for several days with their adhesion maturation. We used the modified system to simultaneously evaluate the stiffness of the cancer cells to which a large deformation was applied and their adhesion strength. The obtained results revealed that the middle phase of tensile stiffness and adhesion force of the microtubule-depolymerized group treated with colchicine (an anti-cancer drug) (stiffness, 13.4 ± 7.5 nN/%; adhesion force, 460.6 ± 258.2 nN) were over two times larger than those of the control group (stiffness, 5.0 ± 3.5 nN/%; adhesion force, 168.2 ± 98.0 nN). Additionally, the same trend was confirmed with the detailed evaluation of cell surface stiffness using an atomic force microscope. Confocal fluorescence microscope observations showed that the stress fibers (SFs) of colchicine-treated cells were aligned in the same direction, and focal adhesions (FAs) of the cells developed around both ends of the SFs and aligned parallel to the developed direction of the SFs. There was a possibility that the microtubule depolymerization by the colchicine treatment induced the development of SFs and FAs and subsequently caused an increment of cell stiffness and adhesion force. From the above results, we concluded the modified system would be applicable to cancer detection and anti-cancer drug efficacy tests.

Folic acid-mediated enhancement of the diagnostic potential of luminescent europium-doped hydroxyapatite nanocrystals for cancer biomaging

Early and accurate cancer diagnosis is crucial for improving patient survival rates. Luminescent nanoparticles have emerged as a promising tool in fluorescence bioimaging for cancer diagnosis. To enhance diagnostic accuracy, ligands promoting endocytosis into cancer cells are commonly incorporated onto nanoparticle surfaces. Folic acid (FA) is one such ligand, known to specifically bind to folate receptors (FR) overexpressed in various cancer cells such as cervical and ovarian carcinoma. Therefore, surface modification of luminescent nanoparticles with FA can enhance both luminescence efficiency and diagnostic accuracy. In this study, luminescent europium-doped hydroxyapatite (EuHAp) nanocrystals were prepared via hydrothermal method and subsequently modified with (3-Aminopropyl)triethoxysilane (APTES) followed by FA to target FR-positive human cervical adenocarcinoma cell line (HeLa) cells. The sequential grafting of APTES and then FA formed a robust covalent linkage between the nanocrystals and FA. Rod-shaped FA-modified EuHAp nanocrystals, approximately 100 nm in size, exhibited emission peaks at 589, 615, and 650 nm upon excitation at 397 nm. Despite a reduction in photoluminescence intensity following FA modification, fluorescence microscopy revealed a remarkable 120-fold increase in intensity compared to unmodified EuHAp, attributed to the enhanced uptake of FA-modified EuHAp. Additionally, confocal microscope observations confirmed the specificity and the internalization of FA-modified EuHAp nanocrystals in HeLa cells. In conclusion, the modification of EuHAp nanocrystals with FA presents a promising strategy to enhance the diagnostic potential of cancer bioimaging probes.

From Antagonism to Enhancement: Triton X-100 Surfactant Affects Phenanthrene Interfacial Biodegradation by Mycobacteria through a Shift in Uptake Mechanisms.

Polycyclic aromatic hydrocarbons (PAHs), as persistent environmental pollutants, often reside in nonaqueous-phase liquids (NAPLs). Mycobacterium sp. WY10, boasting highly hydrophobic surfaces, can adsorb to the oil-water interface, stabilizing the Pickering emulsion and directly accessing PAHs for biodegradation. We investigated the impact of Triton X-100 (TX100) on this interfacial uptake of phenanthrene (PHE) by Mycobacteria, using n-tetradecane (TET) and bis-(2-ethylhexyl) phthalate (DEHP) as NAPLs. Interfacial tension, phase behavior, and emulsion stability studies, alongside confocal laser scanning microscopy and electron microscope observations, unveiled the intricate interplay. In surfactant-free systems, Mycobacteria formed stable W/O Pickering emulsions, directly degrading PHE within the NAPLs because of their intimate contact. Introducing low-dose TX100 disrupted this relationship. Preferentially binding to the cells, the surfactant drastically increased the cell hydrophobicity, triggering desorption from the interface and phase separation. Consequently, PAH degradation plummeted due to hindered NAPL access. Higher TX100 concentrations flipped the script, creating surfactant-stabilized O/W emulsions devoid of interfacial cells. Surprisingly, PAH degradation remained efficient. This paradox can be attributed to NAPL emulsification, driven by the surfactant, which enhanced mass transfer and brought the substrate closer to the cells, despite their absence at the interface. This study sheds light on the complex effect of surfactants on Mycobacteria and PAH uptake, revealing an antagonistic effect at low concentrations that ultimately leads to enhanced degradation through emulsification at higher doses. These findings offer valuable insights into optimizing bioremediation strategies in PAH-contaminated environments.

In Vivo and In Vitro Interactions between Exopolysaccharides from Bacillus thuringensis HD270 and Vip3Aa11 Protein.

Bacillus thuringiensis (Bt) secretes the nutritional insecticidal protein Vip3Aa11, which exhibits high toxicity against the fall armyworm (Spodoptera frugiperda). The Bt HD270 extracellular polysaccharide (EPS) enhances the toxicity of Vip3Aa11 protoxin against S. frugiperda by enhancing the attachment of brush border membrane vesicles (BBMVs). However, how EPS-HD270 interacts with Vip3Aa11 protoxin in vivo and the effect of EPS-HD270 on the toxicity of activated Vip3Aa11 toxin are not yet clear. Our results indicated that there is an interaction between mannose, a monosaccharide that composes EPS-HD270, and Vip3Aa11 protoxin, with a dissociation constant of Kd = 16.75 ± 0.95 mmol/L. When EPS-HD270 and Vip3Aa11 protoxin were simultaneously fed to third-instar larvae, laser confocal microscopy observations revealed the co-localization of the two compounds near the midgut wall, which aggravated the damage to BBMVs. EPS-HD270 did not have a synergistic insecticidal effect on the activated Vip3Aa11 protein against S. frugiperda. The activated Vip3Aa11 toxin demonstrated a significantly reduced binding capacity (548.73 ± 82.87 nmol/L) towards EPS-HD270 in comparison to the protoxin (34.96 ± 9.00 nmol/L). Furthermore, this activation diminished the affinity of EPS-HD270 for BBMVs. This study provides important evidence for further elucidating the synergistic insecticidal mechanism between extracellular polysaccharides and Vip3Aa11 protein both in vivo and in vitro.

An ATP-binding cassette transporter, OsABCB24, is involved in female gametophyte development and early seed growth in rice.

Female gametogenesis has been rarely studied due to gametophyte lethality and the unavailability of related genetic resources. In this study, we identified a rice ATP-binding cassette transporter, OsABCB24, whose null function displayed a significantly reduced seed setting rate by as much as 94%-100% compared with that of the wild type (WT). The reciprocal cross of WT and mutant plants demonstrated that the female reproductive organs in mutants were functionally impaired. Confocal microscopy observations revealed that, although megasporogenesis remained unaffected in CRISPR/Cas9 osabcb24 mutants, the formation of female gametophytes was interrupted. Additionally, the structure of the syncytial nucleus was impaired during the initial stages of endosperm formation. Histochemical analysis showed that OsABCB24 was preferentially expressed at the conjunction of receptacle and ovary, spanning from the functional megaspore stage to the two-nucleate embryo sac stage. Further, OsABCB24 was identified as an endoplasmic reticulum membrane-localized protein. Notably, the overexpression of OsABCB24 triggered a 1.5- to 2-fold increase in grain production compared to the WT. Our findings showed that OsABCB24 plays a key role in both female gametophyte development and the early development of seeds.

Reactive oxygen species-sensitive chondroitin sulfate A-cholesteryl hemisuccinate micelles for targeted doxorubicin delivery in tumor therapy

The efficient drug release triggered by the tumor microenvironment is challenging in the development of stimuli-sensitive nanomedicine. In this study, we developed novel polymeric micelles sensitive to reactive oxygen species (ROS) for targeted delivery of doxorubicin (DOX) in tumor therapy. Hydrophilic chondroitin sulfate A (CSA) was conjugated with cholesteryl hemisuccinate (CHS) via a ROS-cleavable thioketal (TK) bond. The resulting CSA-TK-CHS could form self-assembled micelles, and DOX-loaded CSA-TK-CHS (CSA-TK-CHS/DOX) micelles displayed a nearly spherical shape with an average hydrodynamic diameter of 324 nm. The bioresponsive DOX release of CSA-TK-CHS/DOX micelles was evaluated in an H2O2-containing phosphate buffer solution (PBS). CSA-TK-CHS/DOX micelles showed higher cellular uptake than free DOX in 4T1 cells by confocal laser scanning microscopy (CLSM) observation. And CSA-TK-CHS/DOX improved DOX cytotoxicity against CD44-overexpressed 4T1 and CT26 tumor cells. Notably, CSA-TK-CHS/DOX micelles displayed significantly stronger potency (P < 0.01) in antitumor activity than DOX·HCl in orthotropic 4T1-bearing Balb/c mice. Overall, these findings indicate that amphiphilic CSA-TK-CHS micelles are a promising targeted and intelligent drug carrier for tumor therapy.

Exploring Citronella's inhibitory mechanism against Listeria monocytogenes and its utilization in preserving cheese

Listeria monocytogenes presents significant risk to human health due to its high resistance and capacity to form toxin-producing biofilms that contaminate food. The objective of this study was to assess the inhibitory effect of citronella aldehyde (CIT) on L. monocytogenes and investigate the underlying mechanism of inhibition. The results indicated that the minimum inhibitory concentration (MIC) and Minimum sterilisation concentration (MBC) of CIT against L. monocytogenes was 2 μL/mL. At this concentration, CIT was able to effectively suppress biofilm formation and reduce metabolic activity. Crystalline violet staining and MTT reaction demonstrated that CIT was able to inhibit biofilm formation and reduce bacterial cell activity. Furthermore, the motility assessment assay revealed that CIT inhibited bacterial swarming and swimming. Scanning electron microscopy (SEM) and laser confocal microscopy (LSCM) observations revealed that CIT had a significant detrimental effect on L. monocytogenes cell structure and biofilm integrity. LSCM also observed that nucleic acids of L. monocytogenes were damaged in the CIT-treated group, along with an increase in bacterial extracellular nucleic acid leakage. The proteomic results also confirmed the ability of CIT to affect the expression of proteins related to processes including metabolism, DNA replication and repair, transcription and biofilm formation in L. monocytogenes. Consistent with the proteomics results are ATPase activity and ATP content of L. monocytogenes were significantly reduced following treatment with various concentrations of CIT. Notably, CIT showed good inhibitory activity against L. monocytogenes on cheese via fumigation at 4 °C.This study establishes a foundation for the potential application of CIT in food safety control.

Cell density and extracellular matrix composition mitigate bacterial biofilm sensitivity to UV-C LED irradiation

Ultraviolet-C light-emitting diodes (UV-C LEDs) are an emerging technology for decontamination applications in different sectors. In this study, the inactivation of bacterial biofilms was investigated by applying an UV-C LED emitting at 280 nm and by measuring both the influence of the initial cell density (load) and presence of an extracellular matrix (biofilm). Two bacterial strains exposing diverging matrix structures and biochemical compositions were used: Pseudomonas aeruginosa and Leuconostoc citreum. UV-C LED irradiation was applied at three UV doses (171 to 684 mJ/cm2) on both surface-spread cells and on 24-h biofilms and under controlled cell loads, and bacterial survival was determined. All surface-spread bacteria, between 105 and 109 CFU/cm2, and biofilms at 108 CFU/cm2 showed that bacterial response to irradiation was dose-dependent. The treatment efficacy decreased significantly for L. citreum surface-spread cells when the initial cell load was high, while no load effect was observed for P. aeruginosa. Inactivation was also reduced when bacteria were grown under a biofilm form, especially for P. aeruginosa: a protective effect could be attributed to abundant extracellular DNA and proteins in the matrix of P. aeruginosa biofilms, as revealed by Confocal Laser Scanning Microscopy observations. This study showed that initial cell load and exopolymeric substances are major factors influencing UV-C LED antibiofilm treatment efficacy.Key points• Bacterial cell load (CFU/cm2) could impact UV-C LED irradiation efficiency• Characteristics of the biofilm matrix have a paramount importance on inactivation• The dose to be applied can be predicted based on biofilm propertiesGraphical

Physicochemical and foam properties of ovalbumin-carboxymethylcellulose mixtures after mild heat treatment: Comparison of electrostatic repulsion and attraction

The effects of electrostatic repulsion (pH6.5) and attraction (pH4.6) between biopolymers on the physicochemical and foam properties of ovalbumin (OVA)/carboxymethylcellulose (CMC) mixtures subjected to mild heat treatment (60–75 °C) were comparatively investigated. It was found that electrostatic binding between OVA and CMC at pH 4.6 resulted in an increase in the peak temperature of thermal denaturation (Td) of OVA from 71.3 °C to 74.4 °C, while no significant effect was observed at pH 6.5. Thus, 60 and 65 °C below Td were chosen to further investigate the effects of heating on the degree of thermal aggregation, protein conformation and foam properties of OVA/CMC mixtures. Confocal laser scanning microscopy observations and rheological results indicated that electrostatic binding between OVA and CMC at pH 4.6 significantly reduced the degree of protein thermal aggregation, while on the contrary, electrostatic repulsion between proteins and polysaccharides at pH 6.5 promoted OVA thermal aggregation. Raman spectroscopic characterisation indicated that mild heat treatment mainly led to a change in the microenvironment polarity of OVA tyrosine and tryptophan residues, as well as an increase in the content of random coil in the amide III band irrespective of pH. Moreover, after heating at 65 °C for 20 min, The foaming ability of the OVA/CMC mixtures at pH 6.5 and 4.6 increased by approximately 32% and 36% respectively compared to pure OVA. These findings have a positive value for enhancing the heat sterilisation strength of egg white proteins.

Hitchhiking of Cas9 with nucleus-localized proteins impairs its controllability and leads to efficient genome editing of NLS-free Cas9

CRISPR/Cas9 is the most commonly used genome editing tool in eukaryotic cells. To modulate Cas9 entry into the nucleus to enable control of genome editing, we constructed a light-controlled CRISPR/Cas9 system to control exposure of the Cas9 protein NLS. Although blue light irradiation was found to effectively control the entry of Cas9 protein into the nucleus with confocal microscopy observation, effective gene editing occurred in controls with next-generation sequencing analysis. To further clarify this phenomenon, a CRISPR/Cas9 editing system without the NLS and a CRISPR/Cas9 editing system containing an NES were also constructed. Interestingly, both Cas9 proteins could achieve effective editing of target sites with significantly reduced off-target effects. Thus, we speculated that other factors might mediate Cas9 entry into the nucleus. However, NLS-free Cas9 was found to produce effective target gene editing even following inhibition of cell mitosis to prevent nuclear import caused by nuclear membrane disassembly. Furthermore, multiple nucleus-localized proteins were found to interact with Cas9, which could mediate the hitchhiking of NLS-free Cas9 into the nucleus. These findings will inform future attempts to construct controllable gene editing systems, as well as provide new insights into the evolution of the nucleus and of compatible protein functions.

Chaperonin TRiC/CCT subunit CCT7 is involved in the replication of canine parvovirus in F81 cells.

Canine parvovirus (CPV) is one of the most common lethal viruses in canines. The virus disease is prevalent throughout the year, with high morbidity and mortality rate, causing serious harm to dogs and the dog industry. Previously, yeast two hybrid method was used to screen the protein chaperonin containing TCP-1 (CCT7) that interacts with VP2. However, the mechanism of interactions between CCT7 and VP2 on CPV replication remains unclear. In this study, we first verified the interaction between CCT7 and viral VP2 proteins using yeast one-to-one experiment and co-immunoprecipitation (CoIP) experiment. Laser confocal microscopy observation showed that CCT7 and VP2 were able to co-localize and were mostly localized in the cytoplasm. In addition, the study of VP2 truncated mutant found that the interaction region of VP2 with CCT7 was located between amino acids 231 and 320. Cycloheximide (CHX) chase experiments showed that CCT7 can improve the stability of VP2 protein. After further regulation of CCT7 expression in F81 cells, it was found that the expression level of VP2 protein was significantly reduced after knocking down CCT7 expression by RNA interference (RNAi) or HSF1A inhibitor, and increased after overexpressing host CCT7. The study reveals the role of VP2 interacting protein CCT7 in the replication process of CPV, which could provide a potential target for the prevention and control of CPV.

Singapore grouper iridovirus VP20 interacts with grouper TBK1 and IRF3 to attenuate the interferon immune response

Singapore grouper iridovirus (SGIV), belonging to genus Ranavirus, family Iridoviridae, is a highly pathogenic agent and causes heavy economic losses in the global grouper aquaculture. Recent studies demonstrated that SGIV infection attenuated antiviral immune and inflammatory response induced by poly (I:C) in vitro. However, little was known about the potential functions of the immune regulatory proteins encoded by SGIV. Here, we identified the detailed roles of VP20 and clarified the potential mechanism underlying its immune regulatory function during SGIV infection. Our results showed that VP20 was an IE gene, and partially co-localized with Golgi apparatus and lysosomes in grouper cells. Overexpression of VP20 enhanced SGIV replication, demonstrated by the increase in the transcription levels of viral core genes and the protein synthesis of MCP. Reporter gene assays showed that SGIV VP20 overexpression significantly reduced the IFN promoter activity induced by poly (I:C), grouper stimulator of interferon genes (EcSTING) and TANK-binding kinase 1 (EcTBK1). Consistently, the transcription levels of IFN related genes were significantly decreased in VP20 overexpressing cells compared to those in control cells. Co-IP assay and confocal microscopy observations indicated that VP20 co-localized and interacted with EcTBK1 and EcIRF3, but not EcSTING. In addition, VP20 was able to degrade EcIRF3 and attenuate the antiviral action of EcIRF3, while had no effect on EcTBK1. Together, SGIV VP20 was speculated to promote viral replication through attenuating the IFN response mediated by TBK1-IRF3 in vitro. Our findings provided new insights into the immune regulatory function of SGIV encoded unknown proteins.

Polyhexamethylene biguanide promotes adaptive cross-resistance to gentamicin in Escherichia coli biofilms.

Antimicrobial resistance is a critical public health issue that requires a thorough understanding of the factors that influence the selection and spread of antibiotic-resistant bacteria. Biocides, which are widely used in cleaning and disinfection procedures in a variety of settings, may contribute to this resistance by inducing similar defense mechanisms in bacteria against both biocides and antibiotics. However, the strategies used by bacteria to adapt and develop cross-resistance remain poorly understood, particularly within biofilms -a widespread bacterial habitat that significantly influences bacterial tolerance and adaptive strategies. Using a combination of adaptive laboratory evolution experiments, genomic and RT-qPCR analyses, and biofilm structural characterization using confocal microscopy, we investigated in this study how Escherichia coli biofilms adapted after 28 days of exposure to three biocidal active substances and the effects on cross-resistance to antibiotics. Interestingly, polyhexamethylene biguanide (PHMB) exposure led to an increase of gentamicin resistance (GenR) phenotypes in biofilms formed by most of the seven E. coli strains tested. Nevertheless, most variants that emerged under biocidal conditions did not retain the GenR phenotype after removal of antimicrobial stress, suggesting a transient adaptation (adaptive resistance). The whole genome sequencing of variants with stable GenR phenotypes revealed recurrent mutations in genes associated with cellular respiration, including cytochrome oxidase (cydA, cyoC) and ATP synthase (atpG). RT-qPCR analysis revealed an induction of gene expression associated with biofilm matrix production (especially curli synthesis), stress responses, active and passive transport and cell respiration during PHMB exposure, providing insight into potential physiological responses associated with adaptive crossresistance. In addition, confocal laser scanning microscopy (CLSM) observations demonstrated a global effect of PHMB on biofilm architectures and compositions formed by most E. coli strains, with the appearance of dense cellular clusters after a 24h-exposure. In conclusion, our results showed that the PHMB exposure stimulated the emergence of an adaptive cross-resistance to gentamicin in biofilms, likely induced through the activation of physiological responses and biofilm structural modulations altering gradients and microenvironmental conditions in the biological edifice.

Antibiofouling polyethersulfone ultrafiltration membranes functionalized with MoS2 nanosheets and self-cleaning with hydrogen peroxide

The development of biofouling mitigation and cleaning strategy is of paramount importance for ultrafiltration (UF) membrane biofouling. In this study, polyethersulfone (PES) UF membrane was functionalized, namely MoS2/PES composite membrane. The MoS2/PES composite membrane exhibited superior antifouling property as evidenced by antibacterial circle, static and dynamic biofouling experiments. The permeate fluxes of the pristine PES and MoS2/PES composite membrane respectively decreased by 76% and 31% at the end of 3-h filtration. After exposure of E. coli to MoS2, inhibited cell growth led to reduced membrane biofouling. The biofilm formation was retarded under the dynamic flow condition, which were further confirmed by Scanning Electron Microscopy (SEM) and Confocal Laser Scanning Microscopy (CLSM) observations. As for the enhanced biofouling removal by cleaning with H2O2, 62% of the flux recovery rate for the MoS2/PES composite membrane was much higher than the pristine PES membrane (24%). The peroxidase-like properties of MoS2 were responsible for the enhancement of the superior cleaning performance. The use of H2O2 to trigger the in-situ catalytic function of the MoS2 nano-enzyme generated more ROS and O2 for inducing bacterial cells death and facilitating detachment of biofilm. This work will be helpful to provide important insights into developing more effective strategies for biofouling control and cleaning using advanced nanomaterials in membrane-based water treatment processes.

Effect of roasting and high-pressure homogenization on texture, rheology, and microstructure of walnut yogurt

The effect of roasting and high-pressure homogenization on the quality of yogurt made from peeled walnut kernels was explored in this study. The G' and G'' values of yogurt made from walnuts roasted at high temperatures were reduced. The water-holding capacity and hardness of walnut yogurt were reduced to 47.73% and 24.22 g, respectively. Increasing the homogenization pressure reduced the particle size of the walnut yogurt to 20.50 μm. Homogenized walnut milk at 150 MPa increased the viscosity, hardness, and consistency of yogurt product from 11.71 to 16.74 Pa.s, from 30.01 to 71.63 g and from 283.17 to 455.24 g·s, respectively. The confocal laser scanning microscope observation demonstrated a reduction in the size of fat and protein micelles in the homogenized yogurt samples, resulting in a compact structure. This study will contribute valuable scientific insights to the advancement of plant-based yogurt quality.

Efficient wastewater disinfection using a novel microwave discharge electrodeless ultraviolet system with ozone at an ultra-low dose

Microwave discharge electrodeless lamp (MDEL) is a novel ultraviolet (UV) light source. Synergistic disinfection using UV light emitted by MDEL (MWUV) coupled with ozone (O3) at an ultra-low dose was investigated. Escherichia coli and Bacillus subtilis were deactivated more effectively by MWUV/O3 than by either MWUV or O3 alone. MWUV/O3 treatment using an O3 concentration of 0.4 mg/L gave an E. coli inactivation rate of 5.52 log. The photoreactivation degree and rate of E. coli were lower after inactivation by MWUV/O3 treatment than after MWUV treatment alone. The maximum photoreactivation rates after the MWUV/O3 and MWUV treatments were 2.90% and 16.08%, respectively. MWUV/O3 disinfection also inhibited dark resurrection of E. coli and gave a maximum dark resurrection rate of 0.0036%. Electron paramagnetic resonance spectroscopy indicated that more hydroxyl radicals were generated during MWUV/O3 treatment. Scanning electron microscopy and laser confocal scanning microscopy observations indicated that O3 played a key role in breaking down the cell structure. MWUV/O3 treatment gave a good disinfection effect on fecal coliform bacteria in actual domestic wastewater. The results indicated that inactivation of bacteria can be more effectively achieved by MWUV treatment with O3.

Some Considerations on Austenite Grain Growth Kinetics from High‐Temperature Laser Scanning Confocal Microscopy Observations

For more than 20 years, high‐temperature laser scanning confocal microscopy (HT‐LSCM) has become worldwide a proven technique for observing various metallurgical phenomena in situ. HT‐LSCM turns out to be a reliable and effective method for the observation of quantification austenite grain growth kinetics in steel, being highly relevant in casting and steel processing. In the present article, the measurement technique is briefly introduced followed by some basics on austenite grain growth. Finally, selected examples of recent research activities are discussed, including investigations into pure iron and the systems Fe–P, Fe–C, Fe–Al–N, and Fe–C–Nb–N. The influence of P on the grain growth kinetics is remarkable. At 1350 °C, the final grain size decreases from 255 μm in pure iron to 145 μm by adding 0.044 wt% P. In contrast, C enhances the grain growth in specific alloying ranges (0–0.30 wt%C), particularly at elevated temperatures. For a Al content of 0.025 wt% and only 50 wt ppm N, AlN loses the pinning effect above 1150 °C. In case of Nb(C,N), an elevated Nb content (0.085 wt%) provides stabilization of Nb(C,N) at 1150 °C, but at 1250 °C, no pinning of Nb(C,N) is visible anymore.

The seven rice vacuolar sorting receptors localize to prevacuolar compartments

Vacuolar sorting is critically important in plants as it regulates the mobilization of proteins and plays a major role in important agricultural traits like yield and seed protein content. Vacuolar sorting receptors (VSRs) are integral membrane proteins that mediate protein trafficking from the Golgi apparatus to the vacuole via the intermediate membrane-bound prevacuolar compartment (PVC)/multivesicular body (MVB). VSR proteins, such as an 80 kD (BP-80) from pea, also serve as markers for PVC/MVB. Dissecting VSR-mediated protein trafficking pathways may provide ways to enhance agronomic traits and crop yield. Green fluorescence protein (GFP) fusions with the seven Arabidopsis (Arabidopsis thaliana) VSRs were previously shown to localize to PVCs in transgenic tobacco BY-2 cells. The Rice (Oryza sativa) genome contains seven VSRs (OsVSR1-7), but little is known about their subcellular localizations. Here we studied the subcellular localization of OsVSR1-7 b y using a reporter approach, in which GFP-OsVSR1-7 fusions containing the transmembrane domain (TMD) and cytoplasmic tail (CT) of individual OsVSR were expressed in the protoplasts of rice, transgenic tobacco BY-2 cells and transgenic rice plants. Immunofluorescent labelling studies and confocal laser scanning microscope observation demonstrated that the seven OsVSRs are localized to PVCs and form ring-like structures upon wortmannin treatment. Therefore, we have verified the subcellular localization of OsVSR1-7 in this study. The OsVSRs tagged with GFP can serve as PVCs/MVBs markers in rice for the future studies.