Pulsed light treatment enhances starch hydrolysis and improves starch physicochemical properties of germinated brown rice.

BACKGROUND: Recently, germinated brown rice (GBR) has gained substantial attention as a functional food because of its nutritional attributes. Notably, pulsed light technology (PLT) has emerged as a promising tool for enhancing rice germination and, consequently, has improved the nutritional and functional qualities of GBR-derived products. However, further research is required to comprehensively understand the impact of PLT on GBR physicochemical properties. The present study aimed to investigate the stimulating effects of PLT on starch hydrolysis, starch structure and functional properties of GBR. RESULTS: The PLT substantially boosted α-amylase activity during brown rice germination, leading to a 10.9% reduction in total starch content and a 17.3% increase in reducing sugar content, accompanied by elevated free water levels. Structural analysis indicated no changes in starch crystalline types, whereas gelatinization temperature slightly increased. Pasting properties exhibited a significant drop in peak viscosity. Scanning electron microscopy showed surface erosion of starch granules with microstructural changes. Furthermore, correlation analysis established positive links between α-amylase activity, reducing sugar accumulation, starch structure and functional properties in GBR. CONCLUSION: The present study demonstrates that PLT enhanced the physicochemical properties of GBR starch, significantly improving the stability of GBR products, thereby contributing to expanded applicability of rice starch in the food industry. © 2023 Society of Chemical Industry.

Genomic analysis of a novel Aeromonas veronii phage pAEv1810, belonging to the genus Petsuvirus.

Phage-derived therapies are promising treatments in the fight against bacterial diseases as an alternative strategy nowadays. Species of Aeromonas veronii is an important pathogen causing freshwater fish diseases, the findings on genomic sequences of their bacteriophages are limited. In this work, a lytic bacteriophage capable of specifically infecting A. veronii strain AEv1810 was characterized at the gene level and was designated as pAEv1810. Transmission electron microscopic observation revealed that pAEv1810 belongs to the family of Myoviridae. The genome of phage pAEv1810 consists of 235,066 bp with 38.4% G + C content. Twenty-three of 249 putative proteins encoded by this phage have known functions, and four tRNA genes were found in phage genome. Phylogenetic analyses of RNA polymerase and Terminase large subunit revealed that phage pAEv1810 is closing to those phages classified to the genus Petsuvirus.

Characterization and genomic analysis of the novel Aeromonas veronii phage pAEv1812.

Bacteria of the species Aeromonas veronii are generally ubiquitous in different aquatic environments and are capable of causing a variety of diseases in aquatic animals. In this work, the phage isolate pAEv1812, which is lytic to A. veronii strain 1810, was characterized and sequenced. Morphological analysis revealed that pAEv1812 has a symmetrical head and a long non-contractile tail. A one-step growth curve analysis showed that the latent period and burst period of pAEv1812 were approximately 40 min and 80 min, respectively, with a burst size of 190 PFU/infected cell. The genome of phage pAEv1812 is 61,461 bp in length with a G+C content of 61.4%. Eighteen of the 75 putative proteins encoded by this phage have known functions, and there were no rRNA and tRNA genes in the genome. Phylogenetic analysis based on amino acid sequences of the major capsid protein and terminase large subunit suggested that phage pAEv1812 should be included as a member of the genus Chivirus.

CD205+ polymorphonuclear myeloid-derived suppressor cells suppress antitumor immunity by overexpressing GLUT3.

Myeloid-derived suppressor cells (MDSCs) are responsible for antitumor immunodeficiency in tumor-bearing hosts. Primarily, MDSCs are classified into 2 groups: monocytic (M)-MDSCs and polymorphonuclear (PMN)-MDSCs. In most cancers, PMN-MDSCs (CD11b+ Ly6Clow Ly6G+ cells) represent the most abundant MDSC subpopulation. However, the functional and phenotypic heterogeneities of PMN-MDSC remain elusive, which delays clinical therapeutic targeting decisions. In the 4T1 murine tumor model, CD11b+ Ly6Glow PMN-MDSCs were sensitive to surgical and pharmacological interventions. By comprehensively analyzing 64 myeloid cell-related surface molecule expression profiles, cell density, nuclear morphology, and immunosuppressive activity, the PMN-MDSC population was further classified as CD11b+ Ly6Glow CD205+ and CD11b+ Ly6Ghigh TLR2+ subpopulations. The dichotomy of PMN-MDSCs based on CD205 and TLR2 is observed in 4T07 murine tumor models (but not in EMT6). Furthermore, CD11b+ Ly6Glow CD205+ cells massively accumulated at the spleen and liver of tumor-bearing mice, and their abundance correlated with in situ tumor burdens (with or without intervention). Moreover, we demonstrated that CD11b+ Ly6Glow CD205+ cells were sensitive to glucose deficiency and 2-deoxy-d-glucose (2DG) treatment. Glucose transporter 3 (GLUT3) knockdown by siRNA significantly triggered apoptosis and reduced glucose uptake in CD11b+ Ly6Glow CD205+ cells, demonstrating the dependence of CD205+ PMN-MDSCs survival on both glucose uptake and GLUT3 overexpression. As GLUT3 has been recognized as a target for the rescue of host antitumor immunity, our results further directed the PMN-MDSC subsets into the CD205+ GLUT3+ subpopulation as future targeting therapy.

[Optimization of culture condition for cultivation of influenza virus A on MDCK cell].

OBJECTIVE: To optimize the cultivation condition of influenza virus A on MDCK cells and to improve the separation rate and the inspected effect of influenza virus. METHODS: The virus specimens which came from three kinds of the influenza virus A, swine-origin H1N1, seasonal H1N1 and seasonal H3N2, were cultivated by MDCK cells. It will make the most suitable condition by means of comparing the different inoculum size (25, 50, 75, 100, 125 and 150 microl/cm2), the different concentration of TPCK-Trypsin (1.0, 1.5, 2.0, 2.5, 3.0 and 3.5 microg/ml) as an addition and the different susceptibility of cell generations for serial passage culture of four generations of these three influenza virus. RESULTS: Throughout the comparison the three kinds of the influenza virus, swine-origin H1N1, seasonal H1N1 and seasonal H3N2, which had the most suitable inoculum size 100,75, 75 microl/cm2 and the most suitable concentration of TPCK-Trypsin as an addition 2.5, 2.0, 2.0 microg/ml and the most suitable generations of cell 1, 2, 2 respectively. Earlier generations of MDCK cells of the swine-origin H1N1 were more susceptible than the other influenza virus A. The hemagglutination inhibition (HI) of swine-origin H1N1 after cultivated was lower than the other of seasonal H1N1 and seasonal H3N2. CONCLUSION: The separating effect of the swine-origin H1N1 virus in the MDCK cell was inferior to the seasonal H1N1 and seasonal H3N2. The culture condition of seasonal H1N1 and seasonal H3N2 were roughly similar.

Enhancing Lactobacillus plantarum delivery: Impact of gluconolactone concentration on high-internal-phase emulsion gels and gastrointestinal viability.

In this study, the impact of Gluconolactone (GDL) concentration on the formation of high-internal-phase emulsion gels (HIPEGs) and the gastrointestinal digestive viability of Lactobacillus plantarum encapsulated within these HIPEGs were demonstrated. Increasing GDL concentrations led to cross-linking of particles at the oil-water interface, thereby stabilizing smaller oil droplets. The addition of GDL to HIPEs results in a significant increase in the secondary structure of SPI, specifically in ß-sheet and ß-turn formations, accompanied by a reduction in α-helix percentage. This alteration enhanced the binding effect of protein on water, leading to changes in intermolecular force. Notably, HIPEGs containing 3.0% GDL demonstrated superior encapsulation efficiency and delivery efficiency, reaching 99.0% and 84.5%, respectively. After 14 d of continuous zebrafishs feeding, the intestinal viable cells count of Lactobacillus plantarum reached 1.18 × 107 CFU/mL. This finding supports the potential use of HIPEGs as a probiotic delivery carrier, effectively enhancing the intestinal colonization rate.

Purification and properties of a novel trypsin inhibitor from ginkgo fruits and its antiproliferative effect in triple-negative breast cancer cells.

A novel low molecular mass ginkgo biloba trypsin inhibitor (GBTI) was isolated from ginkgo fruits (GF) by trypsin inhibitory activity-guided fractionation by using ammonium sulphate precipitation, followed by ultra-filtration, affinity chromatography and RP-HPLC. The molecular mass and amino acid sequence of GBTI was determined using ESI-MS and ESI-MS/MS, respectively. The structure of GBTI was identified as MKNLTVIPPICLKFPN, with a molecular mass of 1826 Da. GBTI was stable in the pH range of 4-8 and in the temperature range of 0-80 °C for 30 min. However, the inhibitory activity of the GBTI reduced when incubated with various metalions (K+, Na+, Fe2+, Mg2+ and Ca2+) . Finally, GBTI exhibited significant antiproliferative effect in human MDA-MB-231 and mouse 4 T-1 triple-negative breast cancer cells and without toxicity to MCF-10A normal breast cells. Our results suggest that GBTI could be exploited as a natural and hyperstable anticancer agent for triple-negative breast cancer patients.

Effect of the Hf content on the microstructure and ferroelectric properties of HfxZr1-xO2 thin films using an all-inorganic aqueous precursor solution.

The synthesis of ferroelectric HfxZr1-xO2 (0 ≤ x ≤ 0.50) thin films by chemical solution deposition (CSD) on the surface of Si (100) substrates using all-inorganic salt precursors, is demonstrated in this study. The effects of the Hf content on the microstructure and ferroelectric properties of the films were investigated. The results showed that as the Hf component increased, the root mean square (RMS) roughness as well as the m-phase proportion gradually improved. Near the main diffraction peak of 30.7°, a phase transition from the orthorhombic to the cubic phase, and then to the tetragonal phase occurred. The best ferroelectric behaviour was obtained in the HfxZr1-xO2 film with a Hf content of 14% after 103 field cycling. The H0.14Z0.86O2 thin film exhibited the highest remanent polarization of 12.1 µC cm-2, accompanied by a relative permittivity of 31.8.

Anti-migration and anti-invasion effects of 2-hydroxy-6-tridecylbenzoic acid is associated with the enhancement of CYP1B1 expression through activating the AMPK signaling pathway in triple-negative breast cancer cells.

2-hydroxy-6-tridecylbenzoic acid is alkylsalicylic acid monomer compound, abundantly existed in the ginkgo biloba extracts, however, the underlying mechanism of its anti-migration and anti-invasion effects in triple-negative breast cancer (TNBC) is not clear. Here, 2-hydroxy-6 -tridecylbenzoic acid inhibited MDA-MB-231 and 4 T-1 cells growth without toxicity to MCF-10A normal breast cells. Meanwhile, 2-hydroxy-6-tridecylbenzoic acid inhibited cells migration and invasion as well as EMT with the increase of E-cadherin expression accompanied by the decrease of N-cadherin, Vimentin, Snail, MMP-2 and MMP-9 expression. The inhibition was further demonstrated by the enhancement of cytochrome P450 (CYP) 1B1 expression through the activation of AMP activated protein kinase (AMPK) in MDA-MB-231 and 4 T-1 cells. Silencing of CYP1B1 and AMPK with siRNA blocked the inhibitory effects of migration and invasion, and reversed the EMT related genes. These findings may provide a novel mechanism of the 2-hydroxy-6-tridecylbenzoic acid as a molecular-targeted therapeutic drug for TNBC patients.

Antiproliferative effect of 2-Hydroxy-6-tridecylbenzoic acid from ginkgo biloba sarcotestas through the aryl hydrocarbon receptor pathway in triple-negative breast cancer cells.

This study aims to isolate the potential antiproliferative and cytotoxic compounds from ginkgo biloba sarcotestas (GBS) and investigates the underlying mechanism in human MDA-MB-231 and mouse 4T-1 triple-negative breast cancer cells. Our results showed that 2-Hydroxy-6-tridecylbenzoic acid was isolated by cytotoxicity-guided fractionation where different fractions were assessed using MTT assay against MDA-MB-231 and 4T-1 cells. Colony formation assay showed that 2-Hydroxy-6-tridecylbenzoic acid significantly inhibited cell proliferation. The inhibition was associated with the enhancement of cytochrome P450 (CYP) 1B1 expression in a dose- and time-dependent manner and no significant change of CYP1A1 expression by qPCR and Western blot assays in MDA-MB-231 and 4T-1 cells. The mechanism was further demonstrated by the activation of aryl hydrocarbon receptor (AhR) pathway with the upregulation of AhR, AhR nuclear translocator (ARNT) and AhR-dependent xenobiotic response elements (XRE) activity. These findings may have implications for development of anticancer agents containing 2-Hydroxy-6-tridecylbenzoic acid as functional additives.

Development of a high efficient promoter finding method based on transient transfection.

In metazoan genome, the mechanism of gene expression regulation between transcriptional regulatory elements and their target gene is spatiotemporal. Active promoters possess many specific chromosomal features, such as hypersensitive to DNaseI and enrichment of specific histone modifications. In this article, we proposed a novel method which possesses a high efficiency to find promoters in vitro. A promoter-trap library was constructed with totally 706 random mouse genomic DNA fragment clones, and 260 promoter-active fragments of the library were screened by transient transfection into 4T1 cells. To demonstrate the accuracy of this promoter finding method, 13 fragments with promoter activities were randomly selected for published DNase-seq and ChIP-seq data analysis, downstream transcripts prediction and expression confirmation. qRT-PCR results showed that six predicted transcription units were successfully amplified in different mouse tissues/cells or in reconstituted mouse mammary tumors. Our results indicate that this promoter finding method can successfully detect the promoter-active fragments and their downstream transcripts.

Development of a high efficient promoter finding method based on transient transfection.

In metazoan genome, the mechanism of gene expression regulation between transcriptional regulatory elements and their target gene is spatiotemporal. Active promoters possess many specific chromosomal features, such as hypersensitive to DNaseI and enrichment of specific histone modifications. In this article, we proposed a novel method which possesses a high efficiency to find promoters in vitro. A promoter-trap library was constructed with totally 706 random mouse genomic DNA fragment clones, and 260 promoter-active fragments of the library were screened by transient transfection into 4T1 cells. To demonstrate the accuracy of this promoter finding method, 13 fragments with promoter activities were randomly selected for published DNase-seq and ChIP-seq data analysis, downstream transcripts prediction and expression confirmation. qRT-PCR results showed that six predicted transcription units were successfully amplified in different mouse tissues/cells or in reconstituted mouse mammary tumors. Our results indicate that this promoter finding method can successfully detect the promoter-active fragments and their downstream transcripts.

Ganoderma lucidum polysaccharide inhibits prostate cancer cell migration via the protein arginine methyltransferase 6 signaling pathway.

Prostate cancer is one of the most common types of malignant tumor of men worldwide and the incidence and mortality rate is gradually increasing. At present, the molecular mechanisms of growth and migration in human prostate cancer have not been completely elucidated. Studies have demonstrated that Ganoderma lucidum polysaccharides (GLP) can inhibit cancer. Therefore the present study investigated the effect and molecular mechanism of GLP on cell growth and migration of LNCaP human prostate cancer cells. LNCaP cells were transfected with either a protein arginine methyltransferase 6 (PRMT6) overexpression plasmid or PRMT6 small interfering (si)RNA. The cell growth and migration, and the expression of PRMT6 signaling­associated proteins, were investigated following treatment with 5 and 20 µg/ml GLP. The results demonstrated that GLP inhibited cell growth, induced cell cycle arrest, decreased PRMT6, cyclin­dependent kinase 2 (CDK2), focal adhesion kinase (FAK) and steroid receptor coactivator, (SRC) expression, and increased p21 expression in LNCaP cells, as determined by using a Coulter counter, flow cytometry, and reverse transcription­quantitative polymerase chain reaction and western blotting, respectively. Furthermore, GLP significantly inhibited cell migration, as determined by Transwell migration and scratch assays, and altered CDK2, FAK, SRC and p21 expression in LNCaP cells transfected with the PRMT6 overexpression plasmid. By contrast, PRMT6 knockdown by siRNA reduced the effect of GLP on cell migration. These results indicate that GLP was effective in inhibiting cell growth, the cell cycle and cell migration, and the suppressive effect of GLP on cell migration may occur via the PRMT6 signaling pathway. Therefore, it is suggested that GLP may act as a tumor suppressor with applications in the treatment of prostate cancer. The results of the present study provide both the preliminary theoretical and experimental basis for the investigation of GLP as a therapeutic agent.

Leaching behaviour and environmental risk assessment of heavy metals from electronic solder in acidified soil.

The leaching behaviour of Sn and Pb elements from eutectic SnPb solder of electronic waste in acidic soil was investigated through acidification with HCl-H2SO4 solution and compared with saline solution. The amounts of Sn and Pb elements leached, when subjected to acidic soil, are higher than those with saline soil. Evidence for the significantly preferential release of Sn into the leachate is provided; the galvanic couple accelerated such preferential release. Surface product analysis reveals the slight damage of SnPb in saline soil. Serious dissolution due to electrochemical reaction and a thick, porous PbSO4 surface layer are observed in acidified soil, suggesting more severe toxicity potential of Pb in soil rather than in water.