Fluorescence assay for aflatoxin B1 based on aptamer-binding triggered DNAzyme activity.

As a kind of mycotoxin, aflatoxin B1 (AFB1), which is often found in agricultural products, poses a threat to human health. Developing a simple sensitive method for AFB1 detection is in great demand. Here, we reported an aptamer-based fluorescence assay for AFB1 detection by using DNAzyme to generate and amplify a signal. We redesigned a pair of DNA sequences, which originated from the anti-AFB1 aptamer and RNA-cleaving DNAzyme 10-23. In the absence of AFB1, the aptamer hybridized with the region of the substrate-binding arm of the DNAzyme, inhibiting the activity of the DNAzyme. In the presence of AFB1, the binding of AFB1 to the aptamer led to the displacement of the DNAzyme from the aptamer. The substrate-binding arm was unblocked, and the activity of the DNAzyme was restored for the hydrolysis of the fluorophore and quencher-labeled substrate, causing a significant fluorescence increase. This assay could detect AFB1 in the dynamic range from 0.98 to 2000 nmol/L with high selectivity, and the detection limit was 0.98 nmol/L. Moreover, the assay was able to detect AFB1 in a complex sample matrix. This work provides a useful tool for the analysis of AFB1.

Plasmonic Nanotrenches with 1 nm Nanogaps for Surface-Enhanced Raman Scattering-Based Screening of His-Tagged Proteins.

This study represents a highly sensitive and selective approach to protein screening using surface-enhanced Raman scattering (SERS) facilitated by octahedral Au nanotrenches (OANTs). OANTs are a novel class of nanoparticles characterized by narrow, trench-like excavations indented into the eight facets of a Au octahedron. This unique configuration maximizes electromagnetic near-field focusing as the gap distance decreases to ∼1 nm. Owing to geometrical characteristics of the OANTs, near-field focusing can be maximized through the confinement and reflectance of light trapped within the trenches. We used Ni ions and molecular linkers to confer selective binding affinity for His-tagged proteins on the surfaces of the OANTs for SERS-based protein screening. Remarkably, SERS-based protein screening with the surface-modified OANTs yielded outstanding screening capabilities: 100% sensitivity and 100% selectivity in distinguishing His-tagged human serum albumin (HSA) from native HSA. This highlights the significantly enhanced protein screening capabilities achieved through the synergistic combination of SERS and the OANTs.

Liver protects neuron viability and electrocortical activity in post-cardiac arrest brain injury.

Brain injury is the leading cause of mortality among patients who survive cardiac arrest (CA). Clinical studies have shown that the presence of post-CA hypoxic hepatitis or pre-CA liver disease is associated with increased mortality and inferior neurological recovery. In our in vivo global cerebral ischemia model, we observed a larger infarct area, elevated tissue injury scores, and increased intravascular CD45+ cell adhesion in reperfused brains with simultaneous hepatic ischemia than in those without it. In the ex vivo brain normothermic machine perfusion (NMP) model, we demonstrated that addition of a functioning liver to the brain NMP circuit significantly reduced post-CA brain injury, increased neuronal viability, and improved electrocortical activity. Furthermore, significant alterations were observed in both the transcriptome and metabolome in the presence or absence of hepatic ischemia. Our study highlights the crucial role of the liver in the pathogenesis of post-CA brain injury.

Shugan Jieyu capsule effects on peripheral blood micro-124, micro-132, and brain-derived neurotrophic factor in patients with mild to moderate depression.

BACKGROUND: To assess the effectiveness of Shugan Jieyu capsules on peripheral blood miR-124, miR-132, and brain-derived neurotrophic factor (BDNF) levels in patients with mild to moderate depression following coronary artery intervention [percutaneous coronary intervention (PCI)] for coronary heart disease. AIM: To evaluate the therapeutic efficacy of Shugan Jieyu capsules and their effects on the peripheral blood levels of miR-124, miR-132, and BDNF in patients with mild to moderate depression following PCI for coronary heart disease. METHODS: Patients with mild-to-moderate depression of the liver-qi stagnation type after PCI for coronary heart disease at the 305th Hospital of the People's Liberation Army were enrolled from June 2022 to November 2023 and randomly assigned to two groups: Experimental (treated with Shugan Jieyu capsules) and control (treated with escitalopram oxalate tablets). This study compared the antidepressant effects of these treatments using 17-item Hamilton Rating Scale for Depression (HAMD-17) scores, metabolic equivalents, low-density lipoprotein cholesterol, BDNF, high-sensitivity C-reactive protein levels, miR-124 and miR-132 levels, distribution of immune-related lymphocyte subsets, and traditional Chinese medicine syndrome scores before and after 6 weeks of treatment. RESULTS: No significant difference was observed in any index between the two groups before treatment (P > 0.05). After treatment, the total efficacy rates were 93.33% and 90.00% in the experimental and control groups, respectively. Experimental group had significantly lower scores for the main and secondary syndromes compared to the control group (P < 0.05). No significant difference was observed in the metabolic equivalents between the two groups before and after treatment (P > 0.05). The levels of low-density lipoprotein cholesterol, high-sensitivity C-reactive protein, and miR-132 were significantly lower, whereas those of miR-124, BDNF, CD3+T lymphocytes, CD3+CD4+T helper lymphocytes, and CD3+CD4+/CD3+CD8+ cells were significantly higher in the experimental group compared to the control group (P < 0.05). The incidence of adverse reactions during experimental group was significantly lower than that in control group (P < 0.05). CONCLUSION: Shugan Jieyu capsules have good efficacy in patients with mild-to-moderate depression after PCI, and its mechanism may contribute to the regulation of miR-124, miR-132, BDNF levels, and lymphoid immune cells.

Enhancing Gene Transfection Efficiency of Star-Shaped Poly(ß-amino ester)s via "Top-down" Hydrolysis.

Gene therapy has emerged as a potent tool for treating a wide range of hereditary and acquired disorders. However, the development of high-performance nonviral gene delivery vectors remains a significant challenge. Here we report the development of a new type of star-shaped poly(ß-amino ester) (SPAE) through a "top-down" hydrolysis approach and demonstrate its exceptional DNA transfection efficiency and safety profiles. Two SPAEs with different monomer combinations are first synthesized using an "arm first" strategy and then hydrolyzed sequentially to produce h-SPAEs with varied chemical compositions and molecular weights. Results demonstrate that hydrolysis significantly influences the physiological characteristics of the resulting h-SPAEs and h-SPAE/DNA polyplexes. Dependent on the chemical composition, h-SPAEs with low to moderate hydrolysis degrees exhibit superior gene transfection efficiency and cell viability across various cell types. Notably, the leading candidate, h-SPAE-1-5h, achieves up to 88.8% gene transfection efficiency, which was 154-257% higher compared to SPAE-1. This study not only establishes an easy-to-operate "top-down" approach for reshaping the topological structure and chemical composition of SPAEs, but also identifies promising candidates for effective gene transfection. This strategy can be applied to other cationic polymers to enhance their gene transfection performance.

The First Case of Intra-portal Islet Implantation During Liver Machine Perfusion Allowing Simultaneous Islet-liver Transplantation in A Human: A New and Safe Treatment for End-stage Liver Disease Combined With Diabetes Mellitus.

OBJECTIVE: Evaluating the safety and efficacy of implanting a liver with islet grafts into patients with end-stage liver disease and diabetes mellitus (DM). BACKGROUND: DM and end-stage liver diseases are significant health concern worldwide, often coexisting and mutually influencing each other. Addressing both diseases simultaneously is paramount. METHODS: We utilized the islet transplantation combined ischemia-free liver transplantation (ITIFLT) technique to treat a patient with hepatocellular carcinoma (HCC) and type 2 diabetes mellitus (T2DM). The liver was procured and preserved using the ischemia-free liver transplantation (IFLT) technique, and during normothermic machine perfusion (NMP), isolated and purified islet grafts were transplanted into the liver through the portal vein. Finally, the liver, incorporating with the transplant islet grafts, was implanted into the recipient without interruption of blood supply. RESULTS: The patient received both liver and islet graft from the same donor. The patient achieved insulin-independence by post-transplant day (PTD) 9, and both liver and islet function remained robust. The patient was discharged on PTD 18 and experienced no surgical or transplantation-related complications during the follow-up period. Furthermore, islet grafts presence was observed in liver biopsies after islet transplantation. CONCLUSIONS: This landmark case marks the inaugural application of ITIFLT in humans, signifying its potential as a promising treatment modality for end-stage liver disease with DM.

Silica-induced ROS in alveolar macrophages and its role on the formation of pulmonary fibrosis via polarizing macrophages into M2 phenotype: a review.

Alveolar macrophages (AMs), the first line against the invasion of foreign invaders, play a predominant role in the pathogenesis of silicosis. Studies have shown that inhaled silica dust is recognized and engulfed by AMs, resulting in the production of large amounts of silica-induced reactive oxygen species (ROS), including particle-derived ROS and macrophage-derived ROS. These ROS change the microenvironment of the AMs where the macrophage phenotype is stimulated to swift from M0 to M1 and/or M2, and ultimately emerge as the M2 phenotype to trigger silicosis. This is a complex process accompanied by various molecular biological events. Unfortunately, the detailed processes and mechanisms have not been systematically described. In this review, we first systematically introduce the process of ROS induced by silica in AMs. Then, describe the role and molecular mechanism of M2-type macrophage polarization caused by silica-induced ROS. Finally, we review the mechanism of pulmonary fibrosis induced by M2 polarized AMs. We conclude that silica-induced ROS initiate the fibrotic process of silicosis by inducing macrophage into M2 phenotype, and that targeted intervention of silica-induced ROS in AMs can reprogram the macrophage polarization and ameliorate the pathogenesis of silicosis.

Construction of a 12-Gene Prognostic Risk Model and Tumor Immune Microenvironment Analysis Based on the Clear Cell Renal Cell Carcinoma Model.

OBJECTIVES: Accurate survival predictions and early interventional therapy are crucial for people with clear cell renal cell carcinoma (ccRCC). METHODS: In this retrospective study, we identified differentially expressed immune-related (DE-IRGs) and oncogenic (DE-OGs) genes from The Cancer Genome Atlas (TCGA) dataset to construct a prognostic risk model using univariate Cox regression and least absolute shrinkage and selection operator (LASSO) analysis. We compared the immunogenomic characterization between the high- and low-risk patients in the TCGA and the PUCH cohort, including the immune cell infiltration level, immune score, immune checkpoint, and T-effector cell- and interferon (IFN)-γ-related gene expression. RESULTS: A prognostic risk model was constructed based on 9 DE-IRGs and 3 DE-OGs and validated in the training and testing TCGA datasets. The high-risk group exhibited significantly poor overall survival compared with the low-risk group in the training (P < 0.0001), testing (P = 0.016), and total (P < 0.0001) datasets. The prognostic risk model provided accurate predictive value for ccRCC prognosis in all datasets. Decision curve analysis revealed that the nomogram showed the best net benefit for the 1-, 3-, and 5-year risk predictions. Immunogenomic analyses of the TCGA and PUCH cohorts showed higher immune cell infiltration levels, immune scores, immune checkpoint, and T-effector cell- and IFN-γ-related cytotoxic gene expression in the high-risk group than in the low-risk group. CONCLUSION: The 12-gene prognostic risk model can reliably predict overall survival outcomes and is strongly associated with the tumor immune microenvironment of ccRCC.

Near Infrared-Triggered Nitric Oxide-Release Nanovesicles with Mild-Photothermal Antibacterial and Immunomodulation for Healing MRSA-Infected Diabetic Wounds.

Bacterial infection-induced excessive inflammation is a major obstacle in diabetic wound healing. Nitric oxide (NO) exhibits significant antibacterial activity but is extremely deficient in diabetes. Hence, a near-infrared (NIR)-triggered NO release system is constructed through codelivery of polyarginine (PArg) and gold nanorods (Au) in an NIR-activatable methylene blue (MB) polypeptide-assembled nanovesicle (Au/PEL-PBA-MB/PArg). Upon NIR irradiation, the quenched MB in the nanovesicles is photoactivated to generate more reactive oxygen species (ROS) to oxidize PArg and release NO in an on-demand controlled manner. With the specific bacterial capture of phenylboronic acid (PBA), NO elevated membrane permeability and boosted bacterial vulnerability in the photothermal therapy (PTT) of the Au nanorods, which is displayed by superior mild PTT antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA) at temperatures < 49.7 °C in vitro. Moreover, in vivo, the antibacterial nanovesicles greatly suppressed the burst of MRSA-induced excessive inflammation, NO relayed immunomodulated macrophage polarization from M1 to M2, and the excessive inflammatory phase is successfully transferred to the repair phase. In cooperation with angiogenesis by NO, tissue regeneration is accelerated in MRSA-infected diabetic wounds. Therefore, nanoplatform has considerable potential for accelerating the healing of infected diabetic wounds.

All-in-One 2D Molecular Crystal Optoelectronic Synapse for Polarization-Sensitive Neuromorphic Visual System.

Neuromorphic visual systems (NVSs) hold the potential to not only preserve but also enhance human visual capabilities. One such augmentation lies in harnessing polarization information from light reflected or scattered off surfaces like bees, which can disclose unique characteristics imperceptible to the human eyes. While creating polarization-sensitive optoelectronic synapses presents an intriguing avenue for equipping NVS with this capability, integrating functions like polarization sensitivity, photodetection, and synaptic operations into a singular device has proven challenging. This integration typically necessitates distinct functional components for each performance metric, leading to intricate fabrication processes and constraining overall performance. Herein, a pioneering linear polarized light sensitive synaptic organic phototransistor (OPT) based on 2D molecular crystals (2DMCs) with highly integrated, all-in-one functionality, is demonstrated. By leveraging the superior crystallinity and molecular thinness of 2DMC, the synaptic OPT exhibits comprehensive superior performance, including a linear dichroic ratio up to 3.85, a high responsivity of 1.47 × 104 A W-1, and the adept emulation of biological synapse functions. A sophisticated application in noncontact fingerprint detection achieves a 99.8% recognition accuracy, further highlights its potential. The all-in-one 2DMC optoelectronic synapse for polarization-sensitive NVS marks a new era for intelligent perception systems.

Structure-Assisted Design of Chitosanase Product Specificity for the Production of High-Degree Polymerization Chitooligosaccharides.

Chitosanases are valuable enzymatic tools in the food industry for converting chitosan into functional chitooligosaccharides (COSs). However, most of the chitosanases extensively characterized produced a low degree of polymerization (DP) COSs (DP = 1-3, LdpCOSs), indicating an imperative for enhancements in the product specificity for the high DP COS (DP >3, HdpCOSs) production. In this study, a chitosanase from Methanosarcina sp. 1.H.T.1A.1 (OUC-CsnA4) was cloned and expressed. Analysis of the enzyme-substrate interactions and the subsite architecture of the OUC-CsnA4 indicated that a Ser49 mutation could modify its interaction pattern with the substrate, potentially enhancing product specificity for producing HdpCOSs. Site-directed mutagenesis provided evidence that the S49I and S49P mutations in OUC-CsnA4 enabled the production of up to 24 and 26% of (GlcN)5 from chitosan, respectively─the wild-type enzyme was unable to produce detectable levels of (GlcN)5. These mutations also altered substrate binding preferences, favoring the binding of longer-chain COSs (DP >5) and enhancing (GlcN)5 production. Furthermore, molecular dynamics simulations and molecular docking studies underscored the significance of +2 subsite interactions in determining the (GlcN)4 and (GlcN)5 product specificity. These findings revealed that the positioning and interactions of the reducing end of the substrate within the catalytic cleft are crucial factors influencing the product specificity of chitosanase.

Unveiling the potential of triphenylphosphine salts in tuning organic room temperature phosphorescence.

Triphenylphosphine (TPP) salt derivatives, with their rich chemistry of core-substitution, have emerged as promising candidates for ultralong room temperature phosphorescence (RTP) materials owing to their distinct molecular structures, high quantum efficiency and exceptional phosphorescence properties. This feature article highlights the vast potential of TPP salt derivatives in tunable RTP properties by exploring some factors such as the alkyl chains, halogen anions, through-space charge transfer states, etc., and recent advancements in multi-level information encryption, high-level anticounterfeiting tags and X-ray imaging applications. We anticipate that this article will assist in directing future analyses based on the mechanisms underlying the RTP behavior of TPP derivatives and offer guidance for the rational design of high-performance RTP materials.

Clinical impact of metagenomic next-generation sequencing of bronchoalveolar lavage fluids for the diagnosis of pulmonary infections in respiratory intensive care unit.

Background: The real-world clinical impact of mNGS on BALF in the respiratory intensive care unit (RICU) is not yet fully understood. Methods: We investigated the clinical impact of mNGS on BALF samples obtained from 92 patients admitted to the RICU over a 2-year period. We utilized both mNGS and culture methods to evaluate the effectiveness of mNGS in diagnosing pulmonary infections. The clinical impact of mNGS were evaluated by the clinician committees. Results: Among the 92 diagnosed patients, 78 cases (84.7 %) were determined to have infectious diseases caused by pathogenic microorganisms, and the bacterial infections constituted the most prevalent diagnostic category. For mixed infection, the most common type was the Pneumocystis jironecii and cytomegalovirus co-infection. The mNGS results had a positive impact on the clinical management of 43 cases (46.7 %). Moreover, 19 cases (44.2 %) of positive clinical impacts were solely based on new diagnoses made possible by mNGS results. These new diagnoses were particularly helpful for identifying rare pathogens, which could not be detected by conventional diagnostic methods. Conclusions: The BALF mNGS has a positive real-world impact in RICU. Clinician committee play a critical role in ensuring the appropriate use of mNGS.

Multitasking Integrated Metasurface for Electromagnetic Wave Modulation with Reflection, Transmission, and Absorption.

Accommodating multiple tasks within a tiny metasurface unit cell without them interfering with each other is a significant challenge. In this paper, an electromagnetic (EM) wave modulation metasurface capable of reflection, transmission, and absorption is proposed. This multitasking capability is achieved through a cleverly designed multi-layer structure comprising an EM Wave Shield Layer (ESL), a Polarization Modulation Layer (PML), and a Bottom Plate Layer (BPL). The functionality can be arbitrarily switched by embedding control materials within the structure. Depending on external excitation conditions, the proposed metasurface can realize reflection-type co-planar polarization to cross-polarization conversion, transmission-type electromagnetically induced transparency-like (EIT-like) modes, and broadband absorption. Notably, all tasks operate approximately within the same operating frequency band, and their performance can be regulated by the intensity of external excitation. Additionally, the operating principle of the metasurface is analyzed through impedance matching, an oscillator coupling model, and surface current distribution. This metasurface design offers a strategy for integrated devices with multiple functionalities.

Isotropic Quantum Griffiths Singularity in Nd_{0.8}Sr_{0.2}NiO_{2} Infinite-Layer Superconducting Thin Films.

This work reports on the emergence of quantum Griffiths singularity (QGS) associated with the magnetic field induced superconductor-metal transition (SMT) in unconventional Nd_{0.8}Sr_{0.2}NiO_{2} infinite layer superconducting thin films. The system manifests isotropic SMT features under both in-plane and perpendicular magnetic fields. Importantly, after scaling analysis of the isothermal magnetoresistance curves, the obtained effective dynamic critical exponents demonstrate divergent behavior when approaching the zero-temperature critical point B_{c}^{*}, identifying the QGS characteristics. Moreover, the quantum fluctuation associated with the QGS can quantitatively explain the upturn of the upper critical field around zero temperature for both the in-plane and perpendicular magnetic fields in the phase boundary of SMT. These properties indicate that the QGS in the Nd_{0.8}Sr_{0.2}NiO_{2} superconducting thin film is isotropic. Moreover, a higher magnetic field gives rise to a metallic state with the resistance-temperature relation R(T) exhibiting lnT dependence among the 2-10 K range and T^{2} dependence of resistance below 1.5 K, which is significant evidence of Kondo scattering. The interplay between isotropic QGS and Kondo scattering in the unconventional Nd_{0.8}Sr_{0.2}NiO_{2} superconductor can illustrate the important role of rare region in QGS and help to uncover the exotic superconductivity mechanism in this system.

Shape-Regulated Motion and Energy Conversion of Polyelectrolyte Membrane Actuators.

Smart actuators hold great potential in soft robotics and sensors, but their movement at the fluid interface is less understood and controlled, hindering their performances and applications in complicated fluids. Here an ethanol-containing polyelectrolyte actuator is prepared that demonstrates excellent actuating performance via the Marangoni effect. These actuators exhibit enduring (17 min), repeatable (50 cycles), and autonomous motion on the water surface. More importantly, the motion of actuators are dependent on their shapes. Polygonal actuators with more edges exhibit round motion attached to walls of containers, while the actuators with few edges move randomly. On the basis of this property, the circular actuators can pass through pipe bends with S-shaped complex geometry. These unique advantages lend the actuators to successful applications in wireless sensing (standard 0-5 V level signals) for locating obstructions inside invisible pipes and continuous energy harvesting (7700 nC per cycle) for micro mechanical energy.

A matching and localization study of iliac bone graft for repair of talar cartilage injury secondary to lateral ankle instability.

Background: To investigate the search for an Iliac-Talar Grafts on the iliac bone that is morphologically matched to a multiplanar injury lesion of the talus; while utilizing a bone-harvesting guide to ensure precise positioning of the Iliac-Talar Grafts. Methods: A total of twenty-two cases with both talar CT data and iliac CT data were collected from January 2019 to June 2023. One case each of talar deformity injury and bone disease were excluded, resulting in a selection of 20 cases. The medial and lateral target repair areas of the talus were formulated, and virtual surgery was performed by using digital orthopedic technology to locate an iliac-talar restoration on the iliac bone that matched the morphology of the multiplanar injury lesion of the talus. 3D chromatographic deviation analysis was used to assess the accuracy of Iliac-Talar Grafts in terms of morphometric matching and positioning, while personalized iliac bone extraction guides were designed to ensure accurate positioning of the Iliac-Talar Grafts. Results: The best fitting point for repairing the medial talar lesion is determined to be medial to the anterior iliac crest, specifically 2.935 ± 0.365 cm posterior to the anterior superior iliac spine, and 2.550 ± 0.559 cm anterior to the valgus-iliac crest point (VICP). Similarly, for the repair of the lateral talar lesion, the ideal position is found to be lateral to the posterior iliac crest, approximately 2.695 ± 0.640 cm posterior to the valgus-iliac crest point (VICP). Utilizing bone extraction guides enables precise positioning for iliac bone extraction. Conclusion: This study utilizes virtual surgery, 3D chromatographic deviation analysis, and guide plate techniques in digital orthopedics to precisely locate the Iliac-Talar Graft on the iliac bone, matching the morphology of the talar lesion; it provides a new solution for cutting the iliac bone implant that matches the the multifaceted talar lesion to be repaired.

Frequent gene mutations and the correlations with clinicopathological features in clear cell renal cell carcinoma: preliminary study based on Chinese population and TCGA database.

BACKGROUND: Large-scale sequencing plays important roles in revealing the genomic map of ccRCC and predicting prognosis and therapeutic response to targeted drugs. However, the relevant clinical data is still sparse in Chinese population. METHODS: Fresh tumor specimens were collected from 66 Chinese ccRCC patients, then the genomic RNAs were subjected to whole transcriptome sequencing (WTS). We comprehensively analyzed the frequently mutated genes from our hospital's cohort as well as TCGA-KIRC cohort. RESULTS: VHL gene is the most frequently mutated gene in ccRCC. In our cohort, BAP1 and PTEN are significantly associated with a higher tumor grade and DNM2 is significantly associated with a lower tumor grade. The mutant type (MT) groups of BAP1 or PTEN, BAP1 or SETD2, BAP1 or TP53, BAP1 or MTOR, BAP1 or FAT1 and BAP1 or AR had a significantly correlation with higher tumor grade in our cohort. Moreover, we identified HMCN1 was a hub mutant gene which was closely related to worse prognosis and may enhance anti-tumor immune responses. CONCLUSIONS: In this preliminary research, we comprehensively analyzed the frequently mutated genes in the Chinese population and TCGA database, which may bring new insights to the diagnosis and medical treatment of ccRCC.

Valorizing Arthrospira cell residues into polysaccharides: Characterization and application in yogurt.

This study aimed to extract and characterize polysaccharides from Arthrospira cell residue and evaluate their application in yogurt. Four Arthrospira polysaccharides (APP-50, APP-60, APP-70, and APP-80) were obtained by different ethanol concentrations. With the increase in ethanol concentration, the component peaks of polysaccharide became less and the components were simpler. The results showed that APP-60 had the highest neutral sugar content and the densest spherical structure. APP-50 had the highest protein content, the strongest antioxidant capacity, the porous structure, and the structure was incomplete. The addition of polysaccharides increased the viscosity, storage modulus, loss modulus, and particle size of yogurt, and improved the stability of yogurt during long-term storage. The microstructure of yogurt with added polysaccharides was tighter and more orderly than that of the control yogurt. This study demonstrated that Arthrospira polysaccharides could be used as functional ingredients to enhance the quality and nutritional value of yogurt.

Correlation of Vanillin-Induced Cytotoxicity with CYP3A-Mediated Metabolic Activation.

Vanillin (VAN) is a common flavoring agent that can cause liver damage when ingested in large amounts. Nevertheless, the precise processes responsible for its toxicity remain obscure. The present research aimed to examine the metabolic activation of VAN and establish a potential correlation between its reactive metabolites and its cytotoxicity. In rat liver microsomes incubated with VAN, reduced glutathione/N-acetylcysteine (GSH/NAC), and nicotinamide adenine dinucleotide phosphate (NADPH), two conjugates formed from GSH and one conjugate derived from NAC were identified. We also discovered one GSH conjugate in both the bile obtained from rats and the rat primary hepatocytes that were subjected to VAN exposure. Additionally, the NAC conjugate exerted in the urine of VAN-treated rats was observed. These results indicate that a quinone intermediate was produced from VAN both in vitro and in vivo. Next, we identified CYP3A as the main enzyme that initiated the bioactive pathway of VAN. After the activity of CYP3A was selectively inhibited by ketoconazole (KTZ), the generation of the GSH conjugate declined in hepatocytes exposed to VAN. Furthermore, the vulnerability to VAN-induced toxicity was alleviated by KTZ in hepatocytes. Thus, we propose that the cytotoxicity of VAN may derive from metabolic activation triggered by CYP3A.