Deep learning-based image analysis predicts PD-L1 status from 18F-FDG PET/CT images in non-small-cell lung cancer.

Background: There is still a lack of clinically validated biomarkers to screen lung cancer patients suitable for programmed dead cell-1 (PD-1)/programmed dead cell receptor-1 (PD-L1) immunotherapy. Detection of PD-L1 expression is invasively operated, and some PD-L1-negative patients can also benefit from immunotherapy; thus, the joint modeling of both deep learning images and clinical features was used to improve the prediction performance of PD-L1 expression in non-small cell lung cancer (NSCLC). Methods: Retrospective collection of 101 patients diagnosed with pathology in our hospital who underwent 18F FDG PET/CT scans, with lung cancer tissue Tumor Propulsion Score (TPS) ≥1% as a positive expression. Lesions were extracted after preprocessing PET/CT images, and using deep learning 3D DenseNet121 to learn lesions in PET, CT, and PET/CT images, 1,024 fully connected features were extracted; clinical features (age, gender, smoking/no smoking history, lesion diameter, lesion volume, maximum standard uptake value of lesions [SUVmax], mean standard uptake value of lesions [SUVmean], total lesion glycolysis [TLG]) were combined for joint modeling based on the structured data Category Embedding Model. Results: Area under a receiver operating characteristic (ROC) curve (AUC) and accuracy of predicting PD-L1 positive for PET, CT, and PET/CT test groups were 0.814 ± 0.0152, 0.7212 ± 0.0861, and 0.90 ± 0.0605, 0.806 ± 0.023, 0.70 ± 0.074, and 0.950 ± 0.0250, respectively. After joint clinical feature modeling, the AUC and accuracy of predicting PD-L1 positive for PET/CT were 0.96 ± 0.00905 and 0.950 ± 0.0250, respectively. Conclusion: This study combines the features of 18F-FDG PET/CT images with clinical features using deep learning to predict the expression of PD-L1 in NSCLC, suggesting that 18F-FDG PET/CT images can be conducted as biomarkers for PD-L1 expression.

Ac-HSP20 regulates autophagy and promotes the encystation of Acanthamoeba castellanii by inhibiting the PI3K/AKT/mTOR signaling pathway.

BACKGROUND: The encystation of Acanthamoeba castellanii has important ecological and medical significance. Blocking encystation is the key to preventing transmission and curing infections caused by A. castellanii. The formation of autophagosomes is one of the most important changes that occur during the encystation of Acanthamoeba. Our previous studies have shown that the heat shock protein 20 of A. castellanii (Ac-HSP20) is involved in its encystation. This study aimed to determine the role and mechanism of Ac-HSP20 in regulating autophagy involved in the encystation of A. castellanii. METHODS: Immunofluorescence assay, western blotting and transmission electron microscopy were used to analyze the dynamic changes in autophagy during the initiation and continuation of encystation. The knockdown of Ac-HSP20 was performed to clarify its regulation of encystation and autophagy and to elucidate the molecular mechanism by which Ac-HSP20 participates in autophagy to promote cyst maturation. RESULTS: The encystation rates and autophagosomes were significantly decreased by treatment with the autophagy inhibitor 3-MA. The autophagy marker LC3B and autophagic lysosomes increased with the induced duration of encystation and reached the maximum at 48 h. The encystation rate, LC3B expression and autophagosomes decreased when Ac-HSP20 was knocked down by siRNA transfection. In addition, the expression levels of Ac-HSP20 and LC3B increased and the expressions of p-AKT and p-mTOR decreased after 48 h of encystation without knockdown. However, the expressions of p-AKT and p-mTOR increased while the expression of LC3B decreased under the knockdown of Ac-HSP20. Furthermore, the protein expression of LC3B increased when the PI3K/AKT/mTOR signaling pathway was inhibited but decreased when the pathway was activated. CONCLUSIONS: The results demonstrated that autophagy is positively correlated with the encystation of A. castellanii, and Ac-HSP20 regulates autophagy to maintain the homeostasis of A. castellanii by inhibiting the PI3K /AKT /mTOR signaling pathway, thus promoting the maturation and stability of encystation.

Inhibitive Mechanism of Loquat Flower Isolate on Tyrosinase Activity and Melanin Synthesis in Mouse Melanoma B16 Cells.

Melanin naturally exists in organisms and is synthetized by tyrosinase (TYR); however, its over-production may lead to aberrant pigmentation and skin conditions. Loquat (Eriobotrya japonica (Thunb.) Lindl.) flowers contain a variety of bioactive compounds, while studies on their suppressive capabilities against melanin synthesis are limited. Loquat flower isolate product (LFP) was obtained by ethanol extraction and resin purification, and its inhibitory efficiency against TYR activity was investigated by enzyme kinetics and multiple spectroscopy analyses. In addition, the impact of LFP on melanin synthesis-related proteins' expression in mouse melanoma B16 cells was analyzed using Western blotting. HPLC-MS/MS analysis indicated that LFP was composed of 137 compounds, of which 12 compounds, including flavonoids (quercetin, isorhamnoin, p-coumaric acid, etc.) and cinnamic acid and its derivatives, as well as benzene and its derivatives, might have TYR inhibitory activities. LFP inhibited TYR activity in a concentration-dependent manner with its IC50 value being 2.8 mg/mL. The inhibition was an anti-competitive one through altering the enzyme's conformation rather than chelating copper ions at the active center. LFP reduced the expression of TYR, tyrosinase-related protein (TRP) 1, and TRP2 in melanoma B16 cells, hence inhibiting the synthesis of melanin. The research suggested that LFP had the potential to reduce the risks of hyperpigmentation caused by tyrosinase and provided a foundation for the utilization of loquat flower as a natural resource in the development of beauty and aging-related functional products.

Physicochemical properties and flavor substances analyses of refined beef tallow with dry fractionation treatment.

Dry fractionation represents a significant technique for separation of diverse fractions from beef tallow. The objective of this study was to undertake a systematic investigation of alterations in physicochemical properties, crystallization behavior, thermal properties, and flavor compounds that occur during the beef tallow dry fractionation process. The solid component yielded at 40, 30, and 15 °C were 44.88%, 33.72%, and 13.04% respectively, with an 8.36% liquid content at 15 °C, which was consistent with the characteristics of saturated fatty acids content. The ß - ß' transformation in the dry fractionation process was clearly revealed by X-ray diffraction. Differential scanning calorimetry curves exhibited alterations in exothermic and endothermic peak, as well as enthalpy. Electronic nose identified short-chain compounds, aldehydes, ketones, and nitrogen-containing substances as flavor compounds. Volatile compounds were quantified using HS-SPME-GC-MS. Overall, dry fractionation produces beef tallow fractionated compounds with diverse physicochemical properties and aromatic-active substances, thereby expanding its potential utilization.

Differential impacts of porous starch versus its octenyl succinic anhydride-modified counterpart on naringin encapsulation, solubilization, and in vitro release.

The aim of this work was to evaluate the potentials of porous starch (PS) and its octenyl succinic anhydride modified product (OSAPS) as efficient carriers for loading naringin (NA), focusing on encapsulation efficiency (EE, the percentage of adsorbed naringin relative to its initial amount), drug loading (DL, the percentage of naringin in the complex), structural alterations, solubilization and in vitro release of NA using unmodified starch (UMS) and NA as controls. Both the pore diameter and SBET value of PS decreased after esterification with OSA, and a thinner strip-shaped NA (∼145 nm) was observed in the OSAPS-NA complex and (∼150 nm) in the PS-NA complex. OSAPS exhibited reduced short-range ordered structure, as indicated by a lower R1047/1022 (0.73) compared to PS (0.77). Meanwhile, lowest crystallinity (12.81 %) of NA was found in OSAPS-NA. OSAPS-NA exhibited higher EE and DL for NA than PS-NA and a significant increase in NA saturated solubility in deionized water (by 11.63-fold) and simulated digestive fluids (by 24.95-fold) compared to raw NA. OSAPS contained higher proportions of slowly digestible starch and exhibited a lower digestion rate compared to PS, resulting in a longer time for NA release from its complex during the digestion.

Recent Advances in Microfluidic-Based Extracellular Vesicle Analysis.

Extracellular vesicles (EVs) serve as vital messengers, facilitating communication between cells, and exhibit tremendous potential in the diagnosis and treatment of diseases. However, conventional EV isolation methods are labor-intensive, and they harvest EVs with low purity and compromised recovery. In addition, the drawbacks, such as the limited sensitivity and specificity of traditional EV analysis methods, hinder the application of EVs in clinical use. Therefore, it is urgent to develop effective and standardized methods for isolating and detecting EVs. Microfluidics technology is a powerful and rapidly developing technology that has been introduced as a potential solution for the above bottlenecks. It holds the advantages of high integration, short analysis time, and low consumption of samples and reagents. In this review, we summarize the traditional techniques alongside microfluidic-based methodologies for the isolation and detection of EVs. We emphasize the distinct advantages of microfluidic technology in enhancing the capture efficiency and precise targeting of extracellular vesicles (EVs). We also explore its analytical role in targeted detection. Furthermore, this review highlights the transformative impact of microfluidic technology on EV analysis, with the potential to achieve automated and high-throughput EV detection in clinical samples.

Nanoscale dihydroartemisinin@zeolitic imidazolate frameworks for enhanced antigiardial activity and mechanism analysis.

An artificial semisynthetic material can be derived from artemisinin (ART) called dihydroartemisinin (DHA). Although DHA has enhanced antigiardial potential, its clinical application is limited because of its poor selectivity and low solubility. The drug's absorption has a direct impact on the cell, and mechanism research is limited to its destruction of the cytoskeleton. In this study, we used the zeolitic imidazolate framework-8 and loaded it with DHA (DHA@Zif-8) to improve its antigiardial potential. DHA@Zif-8 can enhance cellular uptake, increase antigiardial proliferation and encystation, and expand the endoplasmic reticulum compared with the DHA-treated group. We used RNA sequencing (RNA-seq) to investigate the antigiardial mechanism. We found that 126 genes were downregulated and 123 genes were upregulated. According to the KEGG and GO pathway analysis, the metabolic functions in G. lamblia are affected by DHA@Zif-8 NPs. We used real-time quantitative reverse transcription polymerase chain reaction to verify our results using the RNA-seq data. DHA@Zif-8 NPs significantly enhanced the eradication of the parasite from the stool in vivo. In addition, the intestinal mucosal injury caused by G. lamblia trophozoites markedly improved in the intestine. This research provided the potential of utilizing DHA@Zif-8 to develop an antiprotozoan drug for clinical applications.

Characterization and expression profiles of the ZmLBD gene family in Zea mays.

BACKGROUND: The Lateral Organ Boundaries Domain (LBD) gene family is a family of plant-specific transcription factors (TFs) that are widely involved in processes such as lateral organ formation, stress response, and nutrient metabolism. However, the function of LBD genes in maize remains poorly understood. METHODS AND RESULTS: In this study, a total of 49 ZmLBD genes were identified at the genome-wide level of maize, they were classified into nine branches based on phylogenetic relationships, and all of them were predicted to be nuclear localized. The 49 ZmLBD genes formed eight pairs of segmental duplicates, and members of the same branches' members had similar gene structure and conserved motif composition. The promoters of ZmLBD genes contain multiple types of cis-acting elements. In addition, by constructing the regulatory network of ZmLBD and other genes and miRNAs, 12 and 22 ZmLBDs were found to be involved in the gene regulatory network and miRNA regulatory network, respectively. The expression pattern analysis suggests that ZmLBD genes may be involved in different biological pathways, and drought stress induced the expressions of two inbred lines. CONCLUSIONS: The findings enhance our comprehension of the potential roles of the ZmLBD gene family in maize growth and development, which is pivotal for genetic enhancement and breeding efforts pertaining to this significant crop.

Design of a synthetic enzyme cascade for the in vitro fixation of formaldehyde to acetoin.

Formaldehyde (FALD) has gained prominence as an essential C1 building block in the synthesis of valuable chemicals. However, there are still challenges in converting FALD into commodities. Recently, cell-free biocatalysis has emerged as a popular approach for producing such commodities. Acetoin, also known as 3-hydroxy-2-butanone, has been widely used in food, cosmetic, agricultural and the chemical industry. It is valuable to develop a process to produce acetoin from FALD. In this study, a cell-free multi-enzyme catalytic system for the production of acetoin using FALD as the substrate was designed and constructed. It included three scales: FALD utilization pathway, glycolysis pathway and acetoin synthesis pathway. After the optimization of the reaction system, 20.17 mM acetoin was produced from 122 mM FALD, with a yield of 0.165 mol/mol, reaching 99.0% of the theoretical yield. The pathway provides a new approach for high-yield acetoin production from FALD, which consolidates the foundation for the production of high value-added chemicals using cheap one-carbon compounds.

The characterization of the pectin/alginate nanoparticle for encapsulation of hydroxypropyl-ß-cyclodextrin-complexed naringin and its effects on cellular uptake and oxidative stress in Caco-2 cells.

Naringin (NR) and hydroxypropyl-ß-cyclodextrin (HPCD) can form a water-soluble complex, but it is unstable. This study aimed to investigate the characterization of the pectin/alginate hydrogel nanoparticles (HNPs) loading HPCD-complexed naringin. The encapsulation efficiency and loading capacity of the HNPs for NR were found to be 79.23 % ± 1.31 % and 23.79 % ± 0.67 %, respectively. HNPs had an average diameter of 409.5 ± 8.5 nm, a PDI of 0.237 ± 0.014, and a zeta-potential of -33.5 ± 0.2. FTIR, XRD, and DSC analysis confirmed that the NR-HPCD complex was embedded into the HNPs. In simulated gastrointestinal digestion, the HNPs exhibited a lower cumulative release rate compared to free NR. In Caco-2 cells, the HNPs were more efficiently transported into the cells. Consequently, the HNPs resulted in a greater decrease in ROS levels, more recovery of mitochondrial membrane potential and higher content of glutathione. This study provided a carrier for encapsulating NR, making it possible for use in food or functional food.

The Progressive Utilization of Ponkan Peel Residue for Regulating Human Gut Microbiota through Sequential Extraction and Modification of Its Dietary Fibers.

As a by-product of citrus processing, ponkan (Citrus reticulata Blanco, cv. Ponkan) peel residue is a source of high quality dietary fiber (DF). To make a full utilization of this resource and give a better understanding on the probiotic function of its DF, soluble dietary fiber (SDF) and insoluble dietary fiber (IDF) were extracted from ponkan peel residue (after flavonoids were extracted) using an alkaline method, followed by modifications using a composite physical-enzymatic treatment. The in vitro fermentation properties of the modified SDF and IDF (namely, MSDF and MIDF) and their effects on short-chain fatty acids (SCFA) production and changes in the composition of human gut microbiota were investigated. Results showed that MSDF and MIDF both significantly lowered the pH value and enhanced total SCFA content in the broths after fermented for 24 h by fecal inocula (p < 0.05) with better effects found in MSDF. Both MSDF and MIDF significantly reduced the diversity, with more in the latter than the former, and influenced the composition of human gut microbiota, especially increasing the relative abundance of Bacteroidetes and decreasing the ratio of Firmicutes to Bacteroidetes (F/B) value. The more influential microbiota by MSDF were g-Collinsella, p-Actinobacteria and g-Dialister, while those by MIDF were f-Veillonellaceae, c-Negativicutes and f-Prevotellacese. These results suggested that the modified ponkan peel residue DF can be utilized by specific bacteria in the human gut as a good source of fermentable fiber, providing a basis for the exploitation of the citrus by-product.

Melatonin Promotes Mitochondrial Biogenesis and Mitochondrial Degradation in Hepatocytes During Sepsis.

Objective: This study aimed to investigate the protective mechanisms of melatonin in an in vitro model of sepsis-induced hepatocyte injury, specifically focusing on mitophagy and mitochondrial biogenesis. Methods: In this study, we utilized lipopolysaccharide (LPS)-treated AML12 cells to establish an in vitro model of sepsis-induced hepatocyte injury. The effects of melatonin pretreatment were examined through various analyses, including assessments of oxidative stress, inflammation, mitophagy, mitochondrial biogenesis, and adenosine triphosphate (ATP) levels. Results: The results revealed that LPS-treated AML12 cells exhibited elevated levels of tumor necrosis factor (TNF)-α, interleukin (IL)-6 protein, intracellular reactive oxygen species (ROS), and lipid peroxidation, specifically malondialdehyde (MDA). Moreover, the levels of key markers associated with mitophagy, including PTEN-induced putative kinase 1 (PINK1), parkin, and LC3, were significantly increased (P < .05). Similarly, markers of mitochondrial biogenesis, such as peroxisome proliferator-activated receptor-gamma coactivator 1α (PGC-1α), nuclear respiratory factor 1 (NRF1), and mitochondrial transcription factor A (TFAM), were also significantly increased (P < .05). Conversely, superoxide dismutase (SOD) activity and ATP levels were significantly decreased in LPS-treated AML12 cells compared to the control group (P < .05). However, melatonin pretreatment led to a significant decrease in TNF-α and IL-6 protein levels, intracellular ROS, and MDA levels (P < .05), along with a significant increase in SOD activity, ATP levels, and markers of mitophagy and mitochondrial. Conclusions: Our findings demonstrate that melatonin plays a role in regulating mitochondrial quality control in sepsis-induced hepatocytes. It achieves this result by promoting mitophagy and inducing mitochondrial biogenesis, thereby selectively eliminating dysfunctional mitochondria.

Erratum - Transcription factor nuclear factor erythroid 2 p45-related factor 2 (NRF2) ameliorates sepsis-associated acute kidney injury by maintaining mitochondrial homeostasis and improving the mitochondrial function.

This corrects the article published in European Journal of Histochemistry 2022;66:3412. doi: 10.4081/ejh.2022.3412.

Efficient production of acetoin from lactate by engineered Escherichia coli whole-cell biocatalyst.

Acetoin, an important and high-value added bio-based platform chemical, has been widely applied in fields of foods, cosmetics, chemical synthesis, and agriculture. Lactate is a significant intermediate short-chain carboxylate in the anaerobic breakdown of carbohydrates that comprise ~ 18% and ~ 70% in municipal wastewaters and some food processing wastewaters, respectively. In this work, a series of engineered Escherichia coli strains were constructed for efficient production of acetoin from cheaper and abundant lactate through heterogenous co-expression of fusion protein (α-acetolactate synthetase and α-acetolactate decarboxylase), lactate dehydrogenase and NADH oxidase, and blocking acetate synthesis pathways. After optimization of whole-cell bioconversion conditions, the engineered strain BL-11 produced 251.97 mM (22.20 g/L) acetoin with a yield of 0.434 mol/mol in shake flasks. Moreover, a titer of 648.97mM (57.18 g/L) acetoin was obtained in 30 h with a yield of 0.484 mol/mol lactic acid in a 1-L bioreactor. To the best of our knowledge, this is the first report on the production of acetoin from renewable lactate through whole-cell bioconversion with both high titer and yield, which demonstrates the economy and efficiency of acetoin production from lactate. Key Points • The lactate dehydrogenases from different organisms were expressed, purified, and assayed. • It is the first time that acetoin was produced from lactate by whole-cell biocatalysis. • The highest titer of 57.18 g/L acetoin was obtained with high theoretical yield in a 1-L bioreactor.

Microencapsulation with Different Starch-Based Polymers for Improving Oxidative Stability of Cold-Pressed Hickory (Carya cathayensis Sarg.) Oil.

Hickory (Carya cathayensis Sarg.) oil is a nutrient-dense edible woody oil, with its unsaturated fatty acids accounting for more than 90% of total ones, and liable to oxidation spoilage. To efficiently improve its stability and expand its application fields, the microencapsulation of cold-pressed hickory oil (CHO) by the molecular embedding method and freeze-drying technique was performed using malt dextrin (MD), hydroxylpropyl-ß-cyclodextrin (HP-ß-CD), ß-cyclodextrin (ß-CD), or porous starch (PS) as a wall material. Two wall materials and/or their CHO microcapsulates (CHOM) with higher encapsulation efficiencies (EE) were selected to carry out physical and chemical characterizations using laser particle size diffractometer, scanning electron microscopy, Fourier-transform infrared spectroscopy, X-ray diffraction, thermogravimetric analysis, derivative thermogravimetry, and oxidative stability tests. Results indicated ß-CDCHOM and PSCHOM had significantly higher EE values (80.40% and 75.52%) than MDCHOM and HP-ß-CDCHOM (39.36% and 48.32%). The particle sizes of the two microcapsules selected were both widely distributed with their spans being more than 1 µm and a certain degree of polydispersity. Microstructural and chemical characterizations indicated that ß-CDCHOM had comparatively stable structure and good thermal stability compared with PSCHOM. Storage performances under light, oxygen, and temperature showed that ß-CDCHOM was superior to PSCHOM, especially in terms of thermal and oxidative stability. This study demonstrates that ß-CD embedding can be applied to improve the oxidative stability of vegetable oils such as hickory oil and act as a means of preparing functional supplementary material.

Lung ultrasound score as a predictor of ventilator use in preterm infants with dyspnea within 24 h after dhospitalization.

BACKGROUND: Selecting the correct ventilation strategy is crucial for the survival of preterm infants with dyspnea in NICU. Lung ultrasound score (LUSsc) is a potential predictor for respiratory support patterns in preterm infants. METHODS: We prospectively included 857 preterm infants. LUS was performed in the first 2 h after admission, and LUSsc was determined by two specialist sonographers. Participants were divided into two categories according to gestational age (<32+0 weeks and 32+0-36+6 weeks) and randomly divided into a training set and a validation set. There were two main outcomes: invasive and non-invasive respiratory support. In the training set, clinical factors were analyzed to find the best cut-off value of LUSsc, and consistency was verified in the verification set. The choice of invasive respiratory support was based on neonatal mechanical ventilation strategies. RESULTS: Preterm infants with invasive respiratory support had a higher LUSsc, greater use of Pulmonary Surfactant（PS）, and lower Oxygenation Index（OI）、birth weight than those with non-invasive support. In the <32+0 weeks group, the area under the curve (AUC) for the receiver operating characteristic curve plotted with 2-h LUSsc was 0.749 (95% CI: 0.689-0.809), the cut-off point of LUSsc was 8, and the sensitivity and specificity were 74.0% and 68.3%, respectively. In the 32+0-36+6 weeks group, the AUC was 0.863 (95% CI: 0.811-0.911), with a cut-off point of 7. Sensitivity and specificity were 75.3% and 0.836%, respectively. In the validation set, using the actual clinical respiratory support selection results for verification, the validation results showed for the <32+0 weeks group (Kappa value 0.660, P < 0.05, McNemar test P > 0.05) for preterm 32+0-36+6 weeks (Kappa value 0.779, P < 0.05, McNemar test P > 0.05). CONCLUSION: The LUSsc showed good reliability in predicting respiratory support mode for preterm infants with dyspnea. Registered at ClinicalTrials.gov (identifier: chiCTR1900023869).

Modified porous starch for enhanced properties: Synthesis, characterization and applications.

Native starches have low water solubility at room temperature and poor stability, which demand modifications to overcome. Porous starch as a modified one shows enhanced adsorptive efficiency and solubility compared with its native starch. In contrast, some inherent disadvantages exist, such as weak mechanical strength and low thermal resistance. Fortunately, modified porous starches have been developed to perform well in adsorption capacity and stability. Modified porous starch can be prepared by esterification, crosslinking, oxidation and multiple modifications to the porous starch. The characterization of modified porous starch can be achieved through various analytical techniques. Modified porous starch can be utilized as highly efficient adsorbents and encapsulants for various compounds and applied in various fields. This review dealt with the progress in the preparation, structural characterization and application of modified porous starch. The objective is to provide a reference for its development, utilization, and future research directions.

Hypoglycemic effect of the Phellinus baumii extract with α-glucosidase-inhibited activity and its modulation to gut microbiota in diabetic patients.

Phellinus baumii extract (PBE) possesses considerable α-glucosidase-inhibited activity. This study investigated the hypoglycemic effect in vitro and in vivo using a glucose consumption assay in HepG2 cells, intragastric administration for ten weeks in STZ-induced mice, and intestinal flora fermentation in patients with type 2 diabetes to reveal the possible underlying mechanisms. PBE was prepared, including α-glucosidase-inhibited ethanol extract (EE) and aqueous extract (AE). In vitro, PBE promoted glucose consumption and enhanced glycogen content and hexokinase activity but lowered phosphoenolpyruvate carboxylase kinase activity in HepG2 cells. In vivo, PBE treatment significantly reduced the body weight (p < 0.05) and fasting blood glucose levels of diabetic mice (p < 0.01), with the lowest blood glucose level observed in the EE+AE group. Furthermore, the serum insulin levels and insulin resistance index (HOMA) of PBE-treated groups decreased significantly (p < 0.01). Moreover, gene expression levels of the IRS-1/PI3K/AKT pathway were significantly upregulated by PBE treatment (p < 0.01). In vitro fermentation demonstrated that EE significantly inhibited the production of H2S and NH3 in the intestinal flora fermentation model in diabetic patients (p < 0.05). In addition, the ratio of Firmicutes to Bacteroidetes was reduced, the growth of Lactobacillus and Prevotella 9 was promoted, and Pseudomonas aeruginosa was inhibited. This study provides new insights and clues for using PBE as a functional food and clinical drug for glycemic control.

Protective effect of amino acids on the stability of bayberry anthocyanins and the interaction mechanism between l-methionine and cyanidin-3-O-glycoside.

The protective effects of three amino acids (l-phenylalanine, l-tryptophan and l-methionine) on the stability of bayberry anthocyanins were investigated. The anthocyanin stability under constant illumination (5000 Lux, 50 Hz) or in the presence of ascorbic acid were evaluated by degradation kinetic parameters, and the interaction between l-methionine and cyanidin-3-O-glucoside (C3G) in a model beverage system was analyzed using Fourier transform infrared spectroscopy, 1H nuclear magnetic resonance, X-ray diffraction, molecular docking, and molecular dynamics simulation. Results indicated that the three amino acids significantly reduced the degradation rate of bayberry anthocyanins (p < 0.05), with the most effect by l-methionine. l-methionine could bind to C3G via hydrogen bonds and Van der Waals forces. This study suggested that l-methionine could well protect anthocyanin against degradation in the aqueous solution and have the potential to be used as a co-pigment to improve the sensory property and extend the shelf life of anthocyanin rich berry products.

Transcription factor nuclear factor erythroid 2 p45-related factor 2 (NRF2) ameliorates sepsis-associated acute kidney injury by maintaining mitochondrial homeostasis and improving the mitochondrial function.

Mitochondrial dysfunction has a role in sepsis-associated acute kidney injury (S-AKI), so the restoration of normal mitochondrial homeostasis may be an effective treatment strategy. Transcription factor nuclear factor erythroid 2 p45-related factor 2 (NRF2) is a main regulator of cell-redox homeostasis, and recent studies reported that NRF2 activation helped to preserve mitochondrial morphology and function under conditions of stress. However, the role of NRF2 in the process of S-AKI is still not well understood. The present study investigated whether NRF2 regulates mitochondrial homeostasis and influences mitochondrial function in S-AKI. We demonstrated activation of NRF2 in an in vitro model: lipopolysaccharide (LPS) challenge of ductal epithelial cells of rat renal tubules (NRK-52e cells), and an in vivo model: cecal ligation and puncture (CLP) of rats. Over-expression of NRF2 attenuated oxidative stress, apoptosis, and the inflammatory response; enhanced mitophagy and mitochondrial biogenesis; and mitigated mitochondrial damage in the in vitro model. In vivo experiments showed that rats treated with an NRF2 agonist had higher adenosine triphosphate (ATP) levels, lower blood urea nitrogen and creatinine levels, fewer renal histopathological changes, and higher expression of mitophagy-related proteins [PTEN-induced putative kinase 1 (PINK1), parkin RBR E3 ubiquitin protein ligase (PRKN), microtubule-associated protein 1 light chain 3 II (LC3 II)] and mitochondrial biogenesis-related proteins [peroxisome proliferator-activated receptor γ coactivator-1 (PGC-1α) and mitochondrial transcription factor A (TFAM)]. Electron microscopy of kidney tissues showed that mitochondrial damage was alleviated by treatment with an NRF2 agonist, and the opposite response occurred upon treatment with an NRF2 antagonist. Overall, our findings suggest that mitochondria have an important role in the pathogenesis of S-AKI, and that NRF2 activation restored mitochondrial homeostasis and function in the presence of this disease. This mitochondrial pathway has the potential to be a novel therapeutic target for the treatment of S-AKI.