NCX1/Ca2+ promotes autophagy and decreases bortezomib activity in multiple myeloma through non-canonical NFκB signaling pathway.

Although bortezomib (BTZ) is the cornerstone of anti-multiple myeloma (MM) therapy, the inevitable primary and secondary drug resistance still seriously affects the prognosis of patients. New treatment strategies are in need. Sodium-calcium exchanger 1 (NCX1) is a calcium-permeable ion transporter on the membrane, and our previous studies showed that low NCX1 confers inferior viability in MM cells and suppressed osteoclast differentiation. However, the effect of NCX1 on BTZ sensitivity of MM and its possible mechanism remain unclear. In this study, we investigated the effect of NCX1 on BTZ sensitivity in MM, focusing on cellular processes of autophagy and cell viability. Our results provide evidence that NCX1 expression correlates with MM disease progression and low NCX1 expression increases BTZ sensitivity. NCX1/Ca2+ triggered autophagic flux through non-canonical NFκB pathway in MM cells, leading to attenuated the sensitivity of BTZ. Knockdown or inhibition of NCX1 could potentiate the anti-MM activity of BTZ in vitro and vivo, and inhibition of autophagy sensitized NCX1-overexpressing MM cells to BTZ. In general, this work implicates NCX1 as a potential therapeutic target in MM with BTZ resistance and provides novel mechanistic insights into its vital role in combating BTZ resistance.

Proteomics analysis of enzyme systems and pathway changes during the moromi fermentation of soy sauce mash.

BACKGROUND: During the brewing of soy sauce, the conversion of multiple substances is driven by various microorganisms and their secreted enzyme systems. Soy sauce mash is an important source of enzyme systems during moromi fermentation, but the changes of enzyme systems in soy sauce mash during moromi fermentation are poorly understood. In order to explore the predominant enzyme systems existing during moromi fermentation and to explain the characteristics of the enzyme system changes, an enzymatic activities assay and 4D-label-free proteomics analysis were conducted on soy sauce mash at different stages of fermentation. RESULTS: The activities of hydrolytic enzymes in soy sauce mash decreased continuously throughout the fermentation process, while most of the characteristic physicochemical substances in soy sauce mash supernatant had already accumulated at the early stage of fermentation. Four hydrolytic enzymes were found to be positively correlated with important physicochemical indexes by principal component analysis and Pearson correlation analysis. The proteomics analysis revealed three highly upregulated enzymes and two enzymes that were present in important metabolic pathways throughout the fermentation process. Furthermore, it was found that Aspergillus oryzae was able to accumulate various nutrients in the soy sauce mash by downregulating most of its metabolic pathways. CONCLUSION: Enzymes present with excellent properties during the moromi fermentation period could be obtained from these results. Meanwhile, the characterization of the metabolic pathways of microorganisms during the moromi fermentation period was revealed. The results provide a basis for more scientific and purposeful improvement of moromi fermentation in the future. © 2024 Society of Chemical Industry.

Submerged Culture of Edible and Medicinal Mushroom Mycelia and Their Applications in Food Products: A Review.

Edible mushrooms have rich nutrition (e.g., proteins, dietary fibers, polysaccharides) and they can be potential sources of important ingredients in food processing. However, the cultivation of mushroom fruiting bodies needs a relatively long time, and they can be easily polluted during the growth process. At the same time, a lot of labor and larger planting areas are also required. As we all know, submerged fermentation is a good way to produce edible mushroom mycelia with less environmental pollution and small footprint, which are also rich in nutrition and bioactive components that are used as dietary supplements or health care products in the food industry. Therefore, it can be considered that the replacement of edible mushroom fruiting bodies with edible mushroom mycelia produced through submerged fermentation has great application potential in food production. At present, most of the research about edible mushroom mycelia focuses on the production of bioactive metabolites in fermentation liquid, but there are few reports that concentrate on their applications in food. This paper reviews the research progress of submerged culture of edible mushroom mycelia and their applications in food products.

[68Ga]Pentixafor PET/CT for staging and prognostic assessment of newly diagnosed multiple myeloma: comparison to [18F]FDG PET/CT.

PURPOSE: To evaluate the prognostic performance of [68Ga]Pentixafor PET/CT at baseline for staging of patients with newly diagnosed multiple myeloma (MM) and to compare it with [18F]FDG PET/CT and the Revised-International Staging System (R-ISS). METHODS: Patients who underwent [68Ga]Pentixafor and [18F]FDG PET/CT imaging were retrospectively included. Patient staging was performed according to the Durie-Salmon PLUS staging system based on [68Ga]Pentixafor PET/CT and [18F]FDG PET/CT images, and the R-ISS. Progression-free survival (PFS) at patient follow-up was estimated using the Kaplan-Meier estimator and compared using the log-rank test. Area under the receiver operating characteristic curve (AUC) was calculated to assess predictive performance. RESULTS: Fifty-five MM patients were evaluated. Compared with [18F]FDG PET, [68Ga]Pentixafor PET detected 25 patients as the same stage, while 26 patients were upstaged and 4 patients were downstaged (P = 0.001). After considering the low-dose CT data, there was no statistically significant difference in the number of patients classified in each stage using [68Ga]Pentixafor PET/CT and [18F]FDG PET/CT (P = 0.091). [68Ga]Pentixafor PET/CT-based staging discriminated PFS outcomes in patients with different disease stages (stage I vs. stage II, stage I vs. stage III, and stage II vs. stage III; all P < 0.05), whereas for [18F]FDG PET/CT, there was only a difference in median PFS between stage I and III (P = 0.021). When staged by R-ISS, the median PFS for stage III was significantly lower than that for stage I and II (P = 0.008 and 0.035, respectively). When predicting 2-year PFS based on staging, the AUC of [68Ga]Pentixafor PET/CT was significantly higher than that of [68Ga]Pentixafor PET (0.923 vs. 0.821, P = 0.002), [18F]FDG PET (0.923 vs. 0.752 P = 0.002), and R-ISS (0.923 vs. 0.776, P = 0.005). CONCLUSIONS: [68Ga]Pentixafor PET/CT-based staging possesses substantial potential to predict disease progression in newly diagnosed MM patients.

Ganoderma Fusions with High Yield of Ergothioneine and Comparative Analysis of Its Genomics.

Ergothioneine (EGT), an exceptional antioxidant found ubiquitously across diverse living organisms, plays a pivotal role in various vital physiological regulatory functions. Its principal natural sources are mushrooms and animal liver tissues. Ganoderma spp., a traditional Chinese food and medicinal mushroom, boasts high concentrations of EGT. To advance the development of novel Ganoderma spp. strains with enhanced EGT yields, we employed an efficient Ganoderma spp. protoplasmic fusion system. Through molecular and biological characterization, we successfully generated seven novel fusion strains. Notably, fusion strain RS7 demonstrated a remarkable increase in mycelial EGT production (12.70 ± 1.85 mg/L), surpassing the parental strains FQ16 and FQ23 by 34.23% and 39.10%, respectively. Furthermore, in the context of the fruiting body, fusion strain RS11 displayed a notable 53.58% enhancement in EGT production (11.24 ± 1.96 mg/L) compared to its parental strains. Genomic analysis of the RS7, the strain with the highest levels of mycelial EGT production, revealed mutations in the gene EVM0005141 associated with EGT metabolism. These mutations led to a reduction in non-productive shunts, subsequently redirecting more substrate towards the EGT synthesis pathway. This redirection significantly boosted EGT production in the RS7 strain. The insights gained from this study provide valuable guidance for the commercial-scale production of EGT and the selective breeding of Ganoderma spp. strains.

The beneficial potential of protein hydrolysates as prebiotic for probiotics and its biological activity: a review.

Probiotics are not only a food supplement, but they have shown great potential in their nutritional, health and therapeutic effects. To maximize the beneficial effects of probiotics, it is commonly achieved by adding prebiotics. Prebiotics primarily comprise indigestible carbohydrates, specific peptides, proteins, and lipids, with oligosaccharides being the most extensively studied prebiotics. However, these rapidly fermenting oligosaccharides have many drawbacks and can cause diarrhea and flatulence in the body. Hence, the exploration of new prebiotic is of great interest. Besides oligosaccharides, protein hydrolysates have been demonstrated to enhance the expression of beneficial properties of probiotics. Consequently, this paper outlines the mechanism underlying the action of protein hydrolysates on probiotics, as well as the advantageous impacts of proteins hydrolysates derived from various food sources on probiotics. In addition, this paper also reviews the currently reported biological activities of protein hydrolysates. The aim is a theoretical basis for the development and implementation of novel prebiotics.

Revised spectral optimization approach to remove surface-reflected radiance for the estimation of remote-sensing reflectance from the above-water method.

The effective sea-surface skylight reflectance (ρ) is an important parameter for removing the contribution of surface-reflected radiance when measuring water-leaving radiance (Lw) using the above-water approach (AWA). Radiative simulations and field measurements show that ρ varies spectrally. To improve the determination of Lw (and then remote sensing reflectance, Rrs) from the AWA, we further developed a wavelength-dependent model for ρ to remove surface-reflected radiance, which is applied with a spectral optimization approach for the determination of Rrs. Excellent agreement was achieved between the AWA-derived and skylight-blocked approach (SBA)-obtained Rrs (coefficient of determination > 0.92, mean absolute percentage deviation < ∼ 11% for Rrs > 0.0005 sr-1), even during high wave conditions. We found that the optimization approach with the new ρ model worked very well for a wide range of water types and observation geometries. For developing remote sensing algorithms and evaluating satellite products, it would be beneficial to apply this approach to current and historical above-water in situ measurements of Rrs to improve the quality of these data. In addition, this approach could also increase the number of useable spectra where previously rendered unusable when processed with a traditional scheme.

Cordyceps militaris polysaccharides modulate gut microbiota and improve metabolic disorders in mice with diet-induced obesity.

BACKGROUND: Cordyceps militaris is an edible and medicinal fungus, and its polysaccharides are among its main pharmacological components. They can display immunomodulation, anti-oxidation, anti-inflammation, anti-hypolipidemic, and other functions. The anti-obesity effect of C. militaris polysaccharides (CMP) is not yet fully understood, however. RESULTS: In this study, a CMP diet intervention was applied over a 4 week period to mice with obesity induced by a high-fat diet (HFD), followed by profiling of obesity-induced dyslipidemia, low-grade inflammation, and gut dysbiosis. The results suggested that CMP could significantly reduce HFD-induced obesity, alleviate obesity-induced hyperlipidemia and insulin resistance, and ameliorate systemic inflammation, showing a promising ability to protect mice from obesity. Further analyses revealed that CMP could regulate obesity-induced gut dysbiosis by restoring the phylogenetic diversity of gut microbiota. It could also increase the relative abundance of short-chain fatty acid (SCFA)-producing bacteria, while down-regulating the level of bacteria that were positively related to the development of obesity. A correlation analysis showed that Helicobacter, Allobaculum, Clostridium XVIII, Parabacteroides, Ligilactobacillus, Faecalibaculum, Adlercreutzia, and Mediterraneibacter were positively related to obese phenotypes. CONCLUSION: This study highlights the potential of CMP as a prebiotic agent to protect obese individuals from metabolic disorders and gut dysbiosis. © 2022 Society of Chemical Industry.

Rheological and microstructural properties of polysaccharide obtained from the gelatinous Tremella fuciformis fungus.

The gelatinous feature of Tremella fuciformis polysaccharide (TFP) has attracted growing interest in its application as a thickening agent in the food industry. This study aims to reveal the microstructure and rheological properties of TFP. Results showed that TFP randomly distributed in aqueous solutions in an irregular worm-like morphology and formed a more extensive entangled network and stiffer chains at higher concentration solutions. The further rheological study indicated that the TFP solutions exhibited a shear-thinning behavior. Multiple results of dynamic oscillation tests confirmed the viscoelastic properties of TFP. Frequency sweep data display that TFP solutions exhibit solid-like behavior at high frequencies, showing the oscillatory behavior of entangled polymers. The temperature sweep demonstrated that the rheological behavior of TFP is thermally reversible. These results enriched the understanding of the rheology-microstructure relationship of TFP solution and were beneficial to expanding the application of TFP in food processing.

Progress in biological activities and biosynthesis of edible fungi terpenoids.

The edible fungi have both edible and medicinal functions, in which terpenoids are one of the most important active ingredients. Terpenoids possess a wide range of biological activities and show great potential in the pharmaceutical and healthcare industries. In this review, the diverse biological activities of edible fungi terpenoids were summarized with emphasis on the mechanism of anti-cancer and anti-inflammation. Subsequently, this review focuses on advances in knowledge and understanding of the biosynthesis of terpenoids in edible fungi, especially in the generation of sesquiterpenes, diterpenes, and triterpenes. This paper is aim to provide an overview of biological functions and biosynthesis developed for utilizing the terpenoids in edible fungi.

A review on recent advances in LED-based non-thermal technique for food safety: current applications and future trends.

Light-emitting diodes (LEDs) is an eco-friendly light source with broad-spectrum antimicrobial activity. Recent studies have extensively been conducted to evaluate its efficacy in microbiological safety and the potential as a preservation method to extend the shelf-life of foods. This review aims to present the latest update of recent studies on the basics (physical, biochemical and mechanical basics) and antimicrobial activity of LEDs, as well as its application in the food industry. The highlight will be focused on the effects of LEDs on different types (bacteria, yeast/molds, viruses) and forms (planktonic cells, biofilms, endospores, fungal toxin) of microorganisms. The antimicrobial activity of LEDs on various food matrices was also evaluated, together with further analysis on the food-related factors that lead to the differences in LEDs efficiency. Besides, the applications of LEDs on the food-related conditions, packaged food, and equipment that could enhance LEDs efficiency were discussed to explore the future trends of LEDs technology in the food industry. Overall, the present review provides important insights for future research and the application of LEDs in the food industry.

Physicochemical and functional properties of Pleurotus geesteranus proteins.

Edible mushroom protein has been regarded as a promising protein source due to its nutritional value and sustainability. In the present study, Pleurotus geesteranus proteins were extracted with alkaline solution and then precipitated with salting out (PPS) and isoelectric point precipitation (PPI), respectively. The influences of precipitation method on the physicochemical and functional properties of these two kinds of proteins were studied. The results showed that both PPS and PPI had a good balance of essential amino acids. These two proteins were mainly consisted of polypeptides with a molecular weight lower than 70 kDa. Using proteome analysis, a number of 772 and 459 protein compositions were identified in PPS and PPI, respectively. Compared to PPS, PPI showed a higher zeta potential, higher surface hydrophobicity, lower content of ß-sheet and ß-turn secondary structure, as well as lower denaturation temperature (Td) and enthalpy change of the denaturation (ΔH). These differences in the physicochemical properties between PPS and PPI resulted in the occurrence of differences in their functional and digestive properties. For example, PPS showed obviously higher protein solubility in water than PPI, especially at natural pH, PPS solution was clear, while PPI showed precipitates. PPI had higher foam capability (FC), lower foaming stability (FS), and lower emulsion stability index (ESI) as compared to PPS. PPI was easier to digest in the pepsin digestion period, while PPS showed a higher nitrogen release after trypsin digestion. These findings on the physicochemical and functional properties of P. geesteranus proteins will help to broaden their applications as protein ingredient in food industry.

Derivation of uncertainty budgets for continuous above-water radiometric measurements along an Atlantic Meridional Transect.

Fiducial reference measurements are in-situ data traceable to metrology standards, with associated uncertainties. This paper presents the methodology used to derive the uncertainty budget for underway, above-water measurements from the Seabird Hyperspectral Surface Acquisition System deployed on an Atlantic Meridional Transect in 2018. The average uncertainty of remote sensing reflectance for clear sky days was ∼ 6% at wavelengths < 490 nm and ∼ 12% at wavelengths > 550 nm. The environmental variability such as sun position, wind speed and skylight distribution caused the greatest uncertainty. The different components of the uncertainty budget are critically assessed to indicate how the measurement procedure could be improved through reducing the principal uncertainty sources.

Amelioration effect of water extract from Ganoderma resinaceum FQ23 solid-state fermentation fungal substance with high-yield ergothioneine on anxiety-like insomnia mice.

Herein, a solid-state fermentation (SSF) system of Ganoderma resinaceum FQ23 with high-yield ergothioneine (EGT) was established, and the amelioration effect of the water extract from its fungal substance on anxiety-like insomnia mice was studied. The content of EGT in the G. resinaceum FQ23 SSF fungal substance increased to 1.146 ± 0.066 mg g-1 DW in the optimization tests. Besides EGT, the common functional components of the water extract from the G. resinaceum FQ23 SSF fungal substance (GSW) were determined, including triterpenoids, polysaccharides, phenols, proteins and amino acids. The animal experiments showed that GSW could alleviate the anxiety-like behavior, improve the antioxidant capacity and protect the organ structure of the anxiety-like insomnia mice. With an increase in the dose of GSW given to the anxiety-like insomnia mice, their serum 5-HT and GABA levels increased, HPA axis hormone levels significantly decreased, BDNF level notably increased, and the response level of the BDNF/CREB signaling pathway was significantly enhanced, indicating that GSW may improve neuroendocrine regulation and neuroprotection in anxiety-like insomnia mice. A 30-times dose of GSW had no acute toxicity in the normal mice. Therefore, the SSF fungal substance of G. resinaceum FQ23 is a potential dietary source for improving sleep. It can be used as a solid drink to help people who are poor sleepers and as a substitute for tea or coffee to help people who are like to drink tea or coffee and cannot sleep.

The in Vitro Fermentation of Cordyceps militaris Polysaccharides Changed the Simulated Gut Condition and Influenced Gut Bacterial Motility and Translocation.

The motility ability of intestinal lipopolysaccharide (LPS)-producing bacteria determines their translocation to the enterohepatic circulation and works as an infectious complication. In this study, the health effects of Cordyceps militaris polysaccharides (CMPs) were re-evaluated based on whether these polysaccharides could affect the motility of gut commensal LPS-producing bacteria and impede their translocation. The results showed that CMP-m fermentation in the gut could change the chemical environment, leading to a decrease in velocity and a shift in the motility pattern. Further study suggested that detachment/fragmentation of flagella, decreased motor forces, and changed chemical conditions might account for this weakened motility. The adhesion and invasion abilities of gut bacteria were also reduced, with lower expression of virulence-related genes. These results indicated that the health regulation effects of CMP-m might be through decreasing the motility of LPS-producing bacteria, hindering their translocation and therefore reducing the LPS level in the enterohepatic circulation.

Multiplex gene precise editing and large DNA fragment deletion by the CRISPR-Cas9-TRAMA system in edible mushroom Cordyceps militaris.

The medicinal mushroom Cordyceps militaris contains abundant valuable bioactive ingredients that have attracted a great deal of attention in the pharmaceutical and cosmetic industries. However, the development of this valuable mushroom faces the obstacle of lacking powerful genomic engineering tools. Here, by excavating the endogenous tRNA-processed element, introducing the extrachromosomal plasmid and alongside with homologous template, we develop a marker-free CRISPR-Cas9-TRAMA genomic editing system to achieve the multiplex gene precise editing and large synthetic cluster deletion in C. militaris. We further operated editing in the synthetases of cordycepin and ergothioneine to demonstrate the application of Cas9-TRAMA system in protein modification, promoter strength evaluation and 10 kb metabolic synthetic cluster deletion. The Cas9-TRAMA system provides a scalable method for excavating the valuable metabolic resource of medicinal mushrooms and constructing a mystical cellular pathway to elucidate the complex cell behaviours of the edible mushroom.

Enhancement of ergothioneine production by discovering and regulating its metabolic pathway in Cordyceps militaris.

BACKGROUND: Cordyceps militaris is a traditional medicinal fungus contains a variety of functional ingredients and has been developed as an important mushroom food recently. Ergothioneine, one of the antioxidative compounds in C. militaris, is benefits on aging-related diseases and therefore became a novel functional food nutritive fortifier. Currently, the main diet source of ergothioneine is mushroom food. However, the mushroom farming faces the problems such as rather low ingredient yield and spontaneous degeneration associated fruiting body that restricts large scale production of ergothioneine. RESULTS: In this study, we excavated the ergothioneine synthetases in mushroom and modified the genes in C. militaris to construct a new ergothioneine synthesis pathway. By further introducing this pathway into C. militaris genome, we succeeded to increase the ingredients' production of engineering strain, the highest amount of ergothioneine and cordycepin were up to 2.5 g/kg dry weight and 2 g/L, respectively. Additionally, the expression of ergothioneine synthetase genes in the shape-mutated degenerative C. militaris could recover the ability of degenerative strain to produce high amount of ingredients, suggesting the metabolic regulation of ergothioneine might release the symptom of mushroom degeneration. CONCLUSION: This study reveals a new pathway to fulfill the market needs of functional mushroom food and food fortifier ergothioneine. It implied the mycelium of C. militaris could be engineered as a novel medicinal mushroom food which could produce higher amount of valuable ingredients.

Fabrication of gel-like emulsions with γ-zein particles using microfluidization: Structure formation and rheological properties.

In this work, γ-zein, which is a type of cysteine rich prolamine in corn, was prepared into particles for fabricating stable gel-like emulsions. The effects of homogenization pressure (0.1-120 MPa) on structure formation and rheological properties of emulsions were systematically studied. Microscopy showed that γ-zein particles provided stabilization at droplet interfaces, and that excess protein particles provided a particle network in continuous phase, and microfluidization significantly decreased droplet size and induced to form droplet clusters and gel-like network. With an increase of homogenization pressure, protein content adsorbed at droplet interfaces and entrapped within gel-like structure of emulsions increased, and more hydrophobic interactions and disulfide bonds between protein particles were formed. As a result, stronger gel strength of emulsions were observed, for example, apparent viscosity, storage modulus (G'), loss modulus (G''), and G'/ G'' crossover strain (γco) increased, the frequency dependency of G' (n, G' ∼ ωn) decreased. Also, Lissajous curves were used to further understand the non-linear viscoelastic behavior of these emulsions. Emulsions prepared without microfluidization (control, 0.1 MPa) showed a weak gel structure with viscous-dominating behavior at high strain, while emulsions prepared with microfluidization showed predominantly elastic behavior at low strain, and plastic-dominating behavior at high strain. These findings showed that cysteine rich protein based- emulsion gels with tunable microstructure and rheological properties can be easily produced by controlling the homogenization pressure.

Overlapping promoter library designed for rational heterogenous expression in Cordyceps militaris.

BACKGROUND: Cordyceps militaris, a kind of edible and medicinal fungus widely accepted in East Asia, has attracted much attention as a potential cell factory for producing adenosine analogs. Despite the rapid development in gene editing techniques and genome modeling, the diversity of DNA elements in C. militaris was too short to achieve rational heterogeneous expression for metabolic engineering studies. RESULTS: In this study, PtrpC, a kind of promoter with a relatively appropriate expression level and small size, was selected as a monomer for promoter library construction. Through in vitro BioBricks assembly, 9 overlapping PtrpC promoters with different copy numbers as well as reporter gene gfp were connected and subsequently integrated into the genome of C. militaris. Both the mRNA transcription level and the expression level of gene gfp gradually increased along with the copy number of the overlapping promoter NPtrpC and peaked at 7. In the meantime, no significant difference was found in either the biomass or morphological characteristic of engineered and wild-type strains. CONCLUSIONS: This study firstly expanded the overlapping promoter strategy used in model microorganism in C. militaris. It was a proof-of-concept in fungi synthetic biology and provide a general method to pushed the boundary of promoter engineering in edible mushroom.

Ergothioneine exhibits longevity-extension effect in Drosophila melanogaster via regulation of cholinergic neurotransmission, tyrosine metabolism, and fatty acid oxidation.

Many studies have demonstrated the protective effect of ergothioneine (EGT), the unique sulfur-containing antioxidant found in mushrooms, on several aging-related diseases. Nevertheless, to date, no single study has explored the potential role of EGT in the lifespan of animal models. We show here that EGT consistently extends fly lifespan in diverse genetic backgrounds and both sexes, as well as in a dose and gender-dependent manner. Additionally, EGT is shown to increases the climbing activity of flies, enhance acetylcholinesterase (AchE) activity, and maintain the ratio of reduced glutathione (GSH) to oxidized glutathione (GSSG)of aged flies. The increase in lifespan by EGT is gut microorganism dependent. We proposed potential mechanisms of lifespan extension in Drosophila by EGT through RNA-seq analysis: preservation of the normal status of the central nervous system via the coordination of cholinergic neurotransmission, tyrosine metabolism, and peroxisomal proteins, regulation of autophagic activity by altering the lysosomal protein CTSD, and the preservation of normal mitochondrial function through controlled substrate feeding into the tricarboxylic acid (TCA) cycle, the major energy-yielding metabolic process in cells.