Effects of saccharide type and extended heating on the Maillard reaction and physicochemical properties of high-solid gelatin gels.

This research delves into the Maillard reaction (MR) in high-solid gelatin-saccharide mixtures consisting of 8% and 72% of allulose, fructose, or fructo-oligosaccharides, which were subjected to varied duration (0-60min) of thermal processing prior to gelation. Physicochemical properties of the gels, including color, chemical composition, protein crosslinking, mechanical strength, in-vitro digestibility and antioxidant activities, were characterized. At pH ∼5.5 and intermediate water activities (0.6-0.7), fast browning was observed through sugar degradation and sugar-amine interactions, which were intensified by prolonged heating. The MR reactivity of saccharides followed: AL > FRU > FOS. Characteristic products (MRPs, e.g., α-dicarbonyls, 5-hydroxymethylfurfural, and advanced glycation end products) were identified, with the spectra of MRPs varying significantly between monosaccharides and oligosaccharides. The MR-induced protein glycation and crosslinking exhibited certain negative impacts on the gel strength and in-vitro protein digestibility. Furthermore, all gelatin-saccharide mixtures exhibited augmented antioxidant properties, with the gelatin-AL mixtures displaying the highest free radical scavenging rates.

Ultrasound-Assisted CRISPRi-Exosome for Epigenetic Modification of α-Synuclein Gene in a Mouse Model of Parkinson's Disease.

Currently, there is a lack of effective treatment for Parkinson's disease (PD). In PD patients, aberrant methylation of SNCA (α-synuclein gene) has been reported and may be a potential therapeutic target. In this study, we established an epigenetic regulation platform based on an exosomal CRISPR intervention system. With the assist of focused ultrasound (FUS) opening the blood-brain barrier, engineered exosomes carrying RVG (rabies viral glycoprotein) targeting peptide, sgRNA (single guide RNA), and dCas9-DNMT3A (named RVG-CRISPRi-Exo) were efficiently delivered into the brain lesions and induced specific methylation of SNCA. In vivo, FUS combined with RVG-CRISPRi-Exo significantly improved motor performance, balance coordination, and neurosensitivity in PD mice, greatly down-regulated the elevation of α-synuclein (α-syn) caused by modeling, rescued cell apoptosis, and alleviated the progression of PD in mice. [18F]-FP-DTBZ imaging suggested that the synaptic function of the nigrostriatal pathway could be restored, which was conducive to the control of motor behavior in PD mice. Pyrosequencing results showed that RVG-CRISPRi-Exo could methylate CpG at specific sites of SNCA, and this fine-tuned editing achieved good therapeutic effects in PD model mice. In vitro, RVG-CRISPRi-Exo down-regulated SNCA transcripts and α-syn expression and relieved neuronal cell damage. Collectively, our findings provide a proof-of-principle for the development of targeted brain nanodelivery based on engineered exosomes and provide insights into epigenetic regulation of brain diseases.

Extracellular Vehicles from Commensal Skin Malassezia restricta Inhibit Staphylococcus aureus Proliferation and Biofilm Formation.

The colonizing microbiota on the body surface play a crucial role in barrier function. Staphylococcus aureus (S. aureus) is a significant contributor to skin infection, and the utilization of colonization resistance of skin commensal microorganisms to counteract the invasion of pathogens is a viable approach. However, most studies on colonization resistance have focused on skin bacteria, with limited research on the resistance of skin fungal communities to pathogenic bacteria. Extracellular vehicles (EVs) play an important role in the colonization of microbial niches and the interaction between distinct strains. This paper explores the impact of Malassezia restricta (M. restricta), the fungus that dominates the normal healthy skin microbiota, on the proliferation of S. aureus by examining the distribution disparities between the two microorganisms. Based on the extraction of EVs, the bacterial growth curve, and biofilm formation, it was determined that the EVs of M. restricta effectively suppressed the growth and biofilm formation of S. aureus. The presence of diverse metabolites was identified as the primary factor responsible for the growth inhibition of S. aureus, specifically in relation to glycerol phospholipid metabolism, ABC transport, and arginine synthesis. These findings offer valuable experimental evidence for understanding microbial symbiosis and interactions within healthy skin.

Microbiomes in pancreatic cancer can be an accomplice or a weapon.

Recently, several investigations have linked the microbiome to pancreatic cancer progression. It is critical to reveal the role of different microbiomes in the occurrence, development, and treatment of pancreatic cancer. The current review summarizes the various microbiota types in pancreatic cancer while updating and supplementing the mechanisms of the representative gut, pancreatic, and oral microbiota, and their metabolites during its pathogenesis and therapeutic intervention. Several novel strategies have been introduced based on the tumor-associated microbiome to optimize the early diagnosis and prognosis of pancreatic cancer. The pros and cons involving different microbiomes in treating pancreatic cancer are discussed. The microbiome-related clinical trials for pancreatic cancer theranostics are outlined. This convergence of cutting-edge knowledge will provide feasible ideas for developing innovative therapies against pancreatic cancer.

Nanomedicine-mediated regulated cell death in cancer immunotherapy.

Immunotherapy has attracted widespread attention in cancer treatment and has achieved considerable success in the clinical treatment of some tumors, but it has a low response rate in most tumors. To achieve sufficient activation of the immune response, significant efforts using nanotechnology have been made to enhance cancer immune response. In recent years, the induction of various regulated cell death (RCD) has emerged as a potential antitumor immuno-strategy, including processes related to apoptosis, autophagy, necroptosis, pyroptosis, ferroptosis, and cuproptosis. In particular, damage-associated molecular patterns (DAMPs) released from the damaged membrane of dying cells act as in situ adjuvants to trigger antigen-specific immune responses by the exposure of an increased antigenicity. Thus, RCD-based immunotherapy offers a new approach for enhancing cancer treatment efficacy. Furthermore, incorporation with multimodal auxiliary therapies in cell death-based immunotherapy can trigger stronger immune responses, resulting in more efficient therapeutic outcome. This review discusses different RCD modalities and summarizes recent nanotechnology-mediated RCDs in cancer immunotherapy.

Effect of Different Degrees of Deep Freezing on the Quality of Snowflake Beef during Storage.

In order to elucidate whether deep freezing could maintain the quality of snowflake beef, three different deep freezing temperatures (-18 °C, -40 °C, and -60 °C) were used in order to evaluate the changes in tissue structures, quality characteristics and spoilage indexes, and their comparative effects on the quality of snowflake beef. Compared to samples frozen at -18 °C, those stored at -40 °C and -60 °C took a shorter time to exceed the maximum ice crystallization zone (significantly reduced by 2-6 h). In terms of short-term storage, samples frozen at -40 °C and -60 °C had better tissue structure and lower drip loss rate than those frozen at -18 °C; significant differences between groups in drip loss were observed between -18 °C and -60 °C. Moreover, a better bright red color and lower shear force were maintained at -40 °C and -60 °C, with significant differences in shear force between the -18 °C group and the other two groups on day 60. Although there were significant effects on the inhibition of lipid and protein oxidation at -40 °C and -60 °C; no significant variation was observed between these two groups throughout storage. A similar phenomenon was found in flavor, with 1-pentanol identified as an important potential indicator of flavor change in snowflake beef during storage. This study demonstrated that -40 °C and -60 °C had favorable impacts on the quality maintenance of snowflake beef compared to -18 °C. These findings provide a theoretical basis for effective stability of snowflake beef quality during frozen storage.

Synergistic gentamicin-photodynamic therapy against resistant bacteria in burn wound infections.

BACKGROUND: Multi-resistant bacteria, partially a result of the abuse of antibiotics, have greatly diminished the effectiveness of antibiotics. The combination of antibiotics with other therapies like antimicrobial photodynamic therapy (aPDT) may provide a useful strategy for fighting resistant bacteria. Here, the synergistic bactericidal effects of toluidine blue (TB)-aPDT and gentamicin (GEN) were evaluated in vitro and in vivo. METHODS: The Post-antibacterial effects were measured at 600 nm (OD600) by a microplate reader. The bacterial envelope and biofilm were observed by a field emission scanning electron microscope. The expression of oxidative stress and Agr system-related genes was analyzed by qRT-PCR after GEN combined with TB-aPDT (GEN&aPDT). Besides, the burn infection model was established to investigate the cloning efficiency of immobilized bacteria, wound healing and inflammatory factors in the lesions. RESULTS: GEN&aPDT could inhibit the growth of Staphylococcus aureus (S. aureus) and multidrug-resistant S. aureus (MDR S. aureus) for up to 15 h, and destroyed the cell envelope and biofilm structure of S. aureus and MDR S. aureus. During the process, ROS played an important role, inducing oxidative stress and downregulating the expression of AgrA, AgrB and PSM in the Agr system, resulting in decreased bacterial virulence and infectivity. In addition, GEN&aPDT cotreatment could effectively promoted wound healing in burn-infected mice by reducing the numbers of bacterial colonization in the wound, decreasing the content of inflammatory factors, and increasing the expression of growth factors. CONCLUSION: The present study confirmed a bactericidal synergy between GEN and aPDT in vitro and in vivo, therein, the oxidative stress exhibited an important role in decreasing bacterial virulence and infectivity, which may bring new ideas for the treatment of bacterial resistance.

TLR 2 induces vascular smooth muscle cell migration through cAMP response element-binding protein-mediated interleukin-6 production.

OBJECTIVE: Migration of vascular smooth muscle cells (VSMCs) from the media into intima contributes to the development of atherosclerosis. Gene deletion experiments implicate a role for toll-like receptor 2 (TLR2) in atherogenesis. However, the underlying mechanisms remain unclear. We postulate that TLR2 promotes VSMC migration by enhancing interleukin (IL)-6 production. METHODS AND RESULTS: Migration assays revealed that TLR2 agonists promoted VSMC migration but not cell proliferation or viability. TLR2 deficiency or inhibition of TLR2 signaling with anti-TLR2 antibody suppressed TLR2 agonist-induced VSMC migration and IL-6 production, which was mediated via p38 mitogen-associated protein kinase and extracellular signal-regulated kinase 1/2 signaling pathways. Neutralizing anti-IL-6 antibodies impaired TLR2-mediated VSMC migration and formation of filamentous actin fiber and lamellipodia. Blockade of p38 mitogen-associated protein kinase or extracellular signal-regulated kinase 1/2 activation inhibited TLR2 agonist pam3CSK4-induced phosphorylation of cAMP response element-binding protein, which regulates IL-6 promoter activity through the cAMP response element site. Moreover, cAMP response element-binding protein small interfering RNA inhibited pam3CSK4-induced IL-6 production and VSMC migration. Additionally, Rac1 small interfering RNA inhibited pam3CSK4-induced VSMC migration but not IL-6 production. CONCLUSIONS: Our results suggest that on ligand binding, TLR2 activates p38 mitogen-associated protein kinase and extracellular signal-regulated kinase 1/2 signaling in VSMCs. These signaling pathways act in concert to activate cAMP response element-binding protein and subsequent IL-6 production, which in turn promotes VSMC migration via Rac1-mediated actin cytoskeletal reorganization.