Autophagic inhibitor ROC-325 ameliorates glomerulosclerosis and podocyte injury via inhibiting autophagic flux in experimental FSGS mice.

BACKGROUND: Autophagy plays an important role in the pathogenesis of focal segmental glomerulosclerosis (FSGS). Podocyte-specific Yes-associated protein (YAP) deletion mice, referred to as YAP-KO mice, is considered a new animal model to study the underlying mechanism of FSGS. ROC-325 is a novel small-molecule lysosomal autophagy inhibitor that is more effective than chloroquine (CQ) and hydroxychloroquine (HCQ) in suppressing autophagy. In this study, we sought to determine the therapeutic benefit and mechanism of action of ROC-325 in YAP-KO mice, an experimental FSGS model. METHODS AND RESULTS: YAP-KO mice were treated with ROC-325 (50 mg/kg, p.o.) daily for one month. Our results revealed that albuminuria, mesangial matrix expension, and focal segmental glomerulosclerosis in YAP-KO mice were significantly attenuated by ROC-325 administration. Transmission electron microscopy and immunofluorescence staining showed that ROC-325 treatment significantly inhibited YAP-KO-induced autophagy activation by decreasing autophagosome-lysosome fusion and increasing LC3A/B and p62/SQSTM. Meanwhile, Immunofluorescence staining revealed that preapplication of ROC-325 in podocyte with YAP-targeted siRNA and mRFP-GFP-LC3 adenovirus markedly suppressed autophagic flux in vitro, suggesting that autophagy intervention may serve as a target for FSGS. CONCLUSIONS: These results showed that the role of autophagic activity in FSGS mice model and ROC-325 could be a novel and promising agent for the treatment of FSGS.

Quantitative proteomics analysis reveals the protective role of S14G-humanin in septic acute kidney injury using 4D-label-free and PRM Approaches.

Mitochondrial dysfunction contributes to septic acute kidney injury (S-AKI), making mitochondrial protection a potential therapeutic strategy. This study investigates the effects of S14G-humanin (HNG) in S-AKI, utilizing 4D-label-free and parallel reaction monitoring (PRM) techniques for proteomic analysis. An S-AKI model was created in male C57BL/6 mice using lipopolysaccharide (LPS) injection, followed by HNG administration. After 24 h, kidney tissues were analyzed for histology, biochemistry, mitochondrial function, and proteomics. HNG treatment improved renal function, reduced tubular injury, and decreased pro-inflammatory cytokines and oxidative stress markers. Proteomic analysis identified 5900 proteins, with 5111 quantifiable. HNG altered the expression of 132 proteins, with 18 selected for PRM validation. Ten of these proteins were linked to key pathways, including fatty acid degradation and PPAR signaling. This study is the first to show HNG's protective effects in S-AKI, providing insights into its mechanisms through advanced proteomic techniques.

Effects of sodium lauryl sulfate and postbiotic toothpaste on oral microecology.

The diversity and delicate balance of the oral microbiome contribute to oral health, with its disruption leading to oral and systemic diseases. Toothpaste includes elements like traditional additives such as sodium lauryl sulfate (SLS) as well as novel postbiotics derived from probiotics, which are commonly employed for maintaining oral hygiene and a healthy oral cavity. However, the response of the oral microbiota to these treatments remains poorly understood. In this study, we systematically investigated the impact of SLS, and toothpaste containing postbiotics (hereafter, postbiotic toothpaste) across three systems: biofilms, animal models, and clinical populations. SLS was found to kill bacteria in both preformed biofilms (mature biofilms) and developing biofilms (immature biofilms), and disturbed the microbial community structure by increasing the number of pathogenic bacteria. SLS also destroyed periodontal tissue, promoted alveolar bone resorption, and enhanced the extent of inflammatory response level. The postbiotic toothpaste favored bacterial homeostasis and the normal development of the two types of biofilms in vitro, and attenuated periodontitis and gingivitis in vivo via modulation of oral microecology. Importantly, the postbiotic toothpaste mitigated the adverse effects of SLS when used in combination, both in vitro and in vivo. Overall, the findings of this study describe the impact of toothpaste components on oral microflora and stress the necessity for obtaining a comprehensive understanding of oral microbial ecology by considering multiple aspects.

Anti-hair loss effect of a shampoo containing caffeine and adenosine.

BACKGROUND: Hair loss is a widespread health problem that affects numerous individuals and is associated with age, lack of sleep, stress, endocrine problems, and other problems. Caffeine exerts various pharmacological effects, particularly after ingestion. The caffeine-induced inhibition of phosphodiesterases can increase intracellular cAMP concentrations, ultimately resulting in stimulatory effects on cell metabolism and proliferation. Hence, caffeine has been confirmed to inhibit hair loss caused by premature termination of the hair growth phase. Adenosine also improves hair loss by stimulating hair growth and thickening hair shafts. However, further empirical evidence is required to comprehensively assess the efficacy of hair loss treatment and prevention using a formulation of caffeine and adenosine in specific proportions in shampoos. OBJECTIVES: This study aimed to evaluate a shampoo with caffeine and adenosine as a daily scalp care product for hair loss in 77 subjects aged 18-60 years. METHODS: The overall and local hair densities were assessed using professional cameras and dermoscopes at different magnifications and distances. Five hairs that came off the participant's head were randomly selected to measure hair diameter. The self-assessment questionnaires were filled on third month of product use. RESULTS: The combination of caffeine and adenosine in the shampoo significantly enhanced hair density compared to that of the baseline. The results revealed a significant reduction in hair loss. The hair diameters of the subjects did not change significantly. Most of the participants (71.05%) were satisfied with their hair after using the product. CONCLUSIONS: Shampoos containing caffeine and adenosine have been demonstrated to exert therapeutic benefits for reducing hair loss.

Engineering Platelet Membrane-Coated Bimetallic MOFs as Biodegradable Nanozymes for Efficient Antibacterial Therapy.

Nanocatalytic-based wound therapeutics present a promising strategy for generating reactive oxygen species (ROS) to antipathogen to promote wound healing. However, the full clinical potential of these nanocatalysts is limited by their low reactivity, limited targeting ability, and poor biodegradability in the wound microenvironment. Herein, a bio-organic nanozyme is developed by encapsulating a FeZn-based bimetallic organic framework (MOF) (MIL-88B-Fe/Zn) in platelet membranes (PM@MIL-88B-Fe/Zn) for antimicrobial activity during wound healing. The introduction of Zn in MIL-88B-Fe/Zn modulates the electronic structure of Fe thus accelerating the catalytic kinetics of its peroxidase-like activity to catalytically generate powerful ROS. The platelet membrane coating of MOF innovatively enhanced the interaction between nanoparticles and the biological environment, further developing bacterial-targeted therapy with excellent antibacterial activity against both gram-positive and gram-negative bacteria. Furthermore, this nanozyme markedly suppressed the levels of inflammatory cytokines and promoted angiogenesis in vivo to effectively treat skin surface wounds and accelerate wound healing. PM@MIL-88B-Fe/Zn exhibited superior biodegradability, favourable metabolism and non-toxic accumulation, eliminating concerns regarding side effects from long-term exposure. The high catalytic reactivity, excellent targeting features, and biodegradability of these nanoenzymes developed in this study provide useful insights into the design and synthesis of nanocatalysts/nanozymes for practical biomedical applications.

Role of TFEB in regulation of the podocyte actin cytoskeleton.

Podocyte injury is linked to the pathogenesis and progression of renal disease. The Transcription Factor EB (TFEB), a master regulator of the autophagy and lysosomal pathways, has been found to exert cell- and tissue-specific biological function. To explore TFEB function and underlying mechanisms in podocytes, a total of 4645 differentially expressed genes (DEGs) were detected in TFEB-knockdown mouse podocytes by transcriptome sequencing. Gene Ontology, Kyoto Encyclopedia of Genes and Genomes, and Ingenuity Pathway Analysis showed that, apart from the enrichment in autophagy and lysosomal pathways, DEGs were enriched in cytoskeleton structure (Actin Cytoskeleton, Focal Adhesion, and Adherens Junction), as well as cytoskeleton regulatory molecular signaling (Hippo and Rho GTPase Signaling). In vitro, TFEB knockdown resulted in podocyte cytoskeletal rearrangement, which was disorganized with cortical distribution of actin filaments. Further, TFEB knockdown decreased mRNA and protein levels of Synaptopodin and led to the rearrangement of Synaptopodin. Inhibition of TFEB decreased mRNA levels for proteins involved in actin cytoskeleton dynamics. Moreover, apoptosis was increased by TFEB knockdown in podocyte. In summary, this study initiated a comprehensive analysis of the role of TFEB in podocyte function and the potential underlying mechanisms, and identified a novel role for TFEB in regulation of the podocyte actin cytoskeleton.

Clinical evidence of the efficacy and safety of a new multi-peptide anti-aging topical eye serum.

BACKGROUND: Skin aging is a complex multifactorial progressive process. With age, intrinsic and extrinsic factors cause the loss of skin elasticity, with the formation of wrinkles, resulting in skin sagging through various pathways. A combination of multiple bioactive peptides could be used as a treatment for skin wrinkles and sagging. OBJECTIVES: This study aimed to evaluate the cosmetic efficacy of a multi-peptide eye serum as a daily skin-care product for improving the periocular skin of women within the ages of 20-45 years. METHODS: The stratum corneum skin hydration and skin elasticity were assessed using a Corneometer CM825 and Skin Elastometer MPA580, respectively. The PRIMOS CR technique based on digital strip projection technology was used for skin image and wrinkle analysis around the "crow's feet" area. Self-assessment questionnaires were filled on Day 14 and 28 of product use. RESULTS: This study included 32 subjects with an average age of 28.5 years. On Day 28, there was a significant decrease in the number, depth, and volume of wrinkles. Skin hydration, elasticity, and firmness increased continuously during the study period, consistent with typical anti-aging claims. A majority of the participants (75.00%) expressed overall satisfaction with their skin appearance after using the product. Most participants noted a visible skin improvement, with an increase in skin elasticity and smoothness, and confirmed the extensibility, applicability, and temperance of the product. No adverse reactions related to product use were observed. CONCLUSIONS: The multi-peptide eye serum uses a multi-targeted mechanism against skin aging to improve the skin appearance, making it an ideal choice for daily skincare.

Growth associated protein 43 deficiency promotes podocyte injury by activating the calmodulin/calcineurin pathway under hyperglycemia.

Podocyte injury is a crucial factor in the pathogenesis of diabetic kidney disease (DKD), and finding potential therapeutic interventions that can mitigate podocyte injury holds significant clinical relevance. This study was to elucidate the role of growth associated protein-43(Gap43) in podocyte injury of high glucose (HG). We confirmed the expression of Gap43 in human glomerulus and found that Gap43 expression was downregulated in podocytes of patients with DKD and HG-treated podocytes in vitro. Gap43 knockdown in podocytes promoted podocyte apoptosis, increased migration ability and decreased nephrin expression, while overexpression of Gap43 markedly suppressed HG-induced injury. Moreover, the increased expression and activity of calcineurin (CaN) were also abrogated by overexpression Gap43 in HG. Pretreatment with a typical CaN inhibitor FK506 in Gap43 knockdown podocytes restored the injury. Mechanistically, co-immunoprecipitation experiments suggested that Gap43 could bind to calmodulin (CaM). Pull-down assay further demonstrated that Gap43 and CaM directly interacts with each other via amino acids 30-52 of Gap43 and amino acids 133-197 of CaM. In addition, we also identified Pax5 as potential transcription inhibitor factor mediating Gap43 expression. In conclusion, the study indicated that the Gap43/CaM-CaN pathway may be exploited as a promising therapeutic target for protecting against podocyte injury in high glucose.

Aberrant NAD synthetic flux in podocytes under diabetic conditions and effects of indoleamine 2,3-dioxygenase on promoting de novo NAD synthesis.

Nicotinamide adenine dinucleotide (NAD) is an essential coenzyme in the kidney. The first step in de novo NAD synthesis is regulated by indoleamine 2,3-dioxygenase (IDO), a tryptophan-catabolizing enzyme. Here, we investigated NAD synthetic flux and NAD levels in podocytes under diabetic conditions. We also studied the effects of IDO overexpression on NAD synthetic flux and high glucose (HG)-induced podocyte injury. NAD synthetases in the de novo, Preiss-Handler and salvage pathways were analyzed using in vivo single-nucleus RNA sequencing datasets (GSE131882) of control and diabetic kidney disease (DKD). The mRNA levels of these NAD synthetases were measured in vitro in HG-treated podocytes. The effects of IDO on NAD synthesis were examined by transducing cultured podocytes with an adenovirus encoding IDO, and apoptosis, podocyte markers and mobility were investigated. Cellular transcriptome analysis revealed that control podocytes had relatively low levels of NAD synthetases. In DKD podocytes, de novo NAD synthetase levels were further downregulated. IDO levels were virtually undetectable and did not increase in DKD. In vitro experiments confirmed aberrant de novo NAD synthetic flux and decreased IDO levels in HG-treated podocytes. Overexpression of IDO promoted NAD de novo synthesis, reduced NAD-bypass metabolic enzyme, increased NAD content and recovered podocyte injury markers under diabetic conditions. Taken together, our findings suggest that the de novo NAD synthetic flux is aberrant in DKD, and IDO promotes de novo NAD synthesis and NAD levels, as well as alleviates injury in HG-treated podocytes.

Enhanced removal of aged and differently functionalized polystyrene nanoplastics using ball-milled magnetic pinewood biochars.

In this study, simple and environmentally friendly magnetic biochars were successfully prepared by ball-milling biochar with Fe3O4 nanoparticles to remove NPs from water. The magnetic biochars synthesized at various pyrolysis temperatures of 300 °C (MBC300), 500 °C (MBC500), and 700 °C (MBC700) were used to eliminate the unmodified (PS), aged under UV radiation (UVPS), amine-modified (PS-NH2) and carboxylate-modified (PS-COOH) polystyrene NPs of 100 nm in size. Results showed that the removal efficiency of MBC300, MBC500, and MBC700 for PS were 43.67, 82.73 and 57.02%, which were 3.01, 5.76, and 3.10 times greater than that of corresponding pristine biochars at the same temperatures, respectively, and the strongest removal efficiency of MBC500 was 95.2% since it has the largest specific surface area and abundant oxygen-containing functional groups. The surface properties of the NPs affected their removal, and the PS-NH2 had the highest removal rate using magnetic biochars. Compared to pristine biochars, the magnetic biochars displayed faster adsorption kinetics. The Langmuir maximum adsorption capacity of magnetic biochars for NPs were 107.7181-229.5772 mg/g, much greater than those of the pristine biochars (55.4602-80.3096 mg/g). Mechanism analysis revealed that the hydrophobicity, electrostatic attraction, H-bonding formation and π-π conjunction between the NPs and MBCs contributed to the adsorption process. This work highlights the promising potential of ball milling to be used as a simple technique for the preparation of magnetic biochar to remove NPs, especially NPs with various surface groups.

Bacillus coagulans Alleviates Intestinal Damage Induced by TiO2 Nanoparticles in Mice on a High-Fat Diet.

Titanium dioxide nanoparticles (TiO2 NPs) are generally added in considerable amounts to food as a food additive. Oral exposure to TiO2 NPs could induce intestinal damage, especially in obese individuals with a high-fat diet. The probiotic Bacillus coagulans (B. coagulans) exhibits good resistance in the gastrointestinal system and is beneficial to intestinal health. In this study, B. coagulans was used to treat intestinal damage caused by TiO2 NPs in high-fat-diet mice via two intervention methods: administration of TiO2 NPs and B. coagulans simultaneously and administration of TiO2 NPs followed by that of B. coagulans. The intervention with B. coagulans was found to reduce the inflammatory response and oxidative stress. A 16S rDNA sequencing analysis revealed that B. coagulans had increased the diversity of gut microbiota and optimized the composition of gut microbiota. Fecal metabolomics analysis indicated that B. coagulans had restored the homeostasis of sphingolipids and amino acid metabolism. The intervention strategy of administering TiO2 NPs followed by B. coagulans was found to be more effective. In conclusion, B. coagulans could alleviate intestinal damage induced by TiO2 NPs in high-fat-diet mice TiO2B. coagulans. Our results suggest a new avenue for interventions against intestinal damage induced by TiO2 NPs.

Perfluorooctane sulfonate (PFOS) enhanced polystyrene particles uptake by human colon adenocarcinoma Caco-2 cells.

As microplastics and nanoplastics (MNPs) are widely distributed in the environment and can be transferred to human body through food chain, their potential impact on human health is of great concern. Perfluorooctane sulfonate (PFOS) is persistent, bioaccumulative and can be adsorbed by MNPs. However, there are few studies on the combined human health effects of MNPs with PFOS. In this study, the effects of polystyrene (PS) particles and PFOS on human colon adenocarcinoma cell Caco-2 were investigated in vitro to explore the combined toxicity from cellular level, and the toxic mechanism was further illustrated. Results showed that the presence of PFOS significantly increased the cell uptake of PS nanoparticles by >30 %, which is related to variations of the surface properties of PS particles, including the decrease of hydration kinetic diameter, the rise of surface potential and the adsorption of hydrophobic PFOS molecules. The toxic effect of PFOS was weakened in the presence of PS particles under low PFOS concentration (10 µg/mL), which is because the bioavailability of PFOS was reduced after adsorption. PS particles with small particle size (20 nm) showed higher cell uptake and ROS production, while PS particles with large particle size (1 µm) led to higher lipid oxidation degree and related membrane damage as well as mitochondrial stress. This study provides the first evaluation of combined toxicity of MNPs and PFOS on human intestinal cells, in order to support the risk assessment of combined pollution of MNPs and PFOS on human health.

The combined effects of nanoplastics and dibutyl phthalate on Streptomyces coelicolor M145.

The environmental and human health risks posed by nanoplastics have attracted considerable attention; however, research on the combined toxicity of nanoplastics and plasticizers is limited. This study analyzed the combined effects of nanoplastics and dibutyl phthalate (DBP) on Streptomyces coelicolor M145 (herein referred to as M145) and its mechanism. The results demonstrated that when the concentration of both nanoplastics and DBP was 1 mg/L, the co-addition was not toxic to M145. When the DBP concentration increased to 5 mg/L, the combined toxicity of 1 mg/L nanoplastics and 5 mg/L DBP reduced when compared to the 5 mg/L DBP treatment group. Similarly, the combined toxicity of 10 mg/L nanoplastics and 1 mg/L DBP on M145 was also lower than that of only 10 mg/L nanoplastics. The co-addition of 10 mg/L nanoplastics and 5 mg/L DBP resulted in the lowest survival rate (41.3%). The key reason for differences in cytotoxicity were variations in the agglomeration of nanoplastics and the adsorption of DBP on nanoplastics. The combination of 10 mg/L nanoplastics and 5 mg/L DBP maximized the production of antibiotics; actinorhodin and undecylprodigiosin yields were 3.5 and 1.8-fold higher than that of the control, respectively. This indicates that the excessive production of antibiotics may be a protective mechanism for bacteria. This study provides a new perspective for assessing the risk of co-exposure to nanoplastics and organic contaminants on microorganisms in nature.

Combined cytotoxicity of polystyrene nanoplastics and phthalate esters on human lung epithelial A549 cells and its mechanism.

Awareness of risks posed by widespread presence of nanoplastics (NPs) and bioavailability and potential to interact with organic pollutants has been increasing. Inhalation is one of the more important pathways of exposure of humans to NPs. In this study, combined toxicity of concentrations of polystyrene NPs and various phthalate esters (PAEs), some of the most common plasticizers, including dibutyl phthalate (DBP) and di-(2-ethyl hexyl) phthalate (DEHP) on human lung epithelial A549 cells were investigated. When co-exposed, 20 µg NPs/mL increased viabilities of cells exposed to either DBP or DEHP and the modulation of toxic potency of DEHP was greater than that of DBP, while the 200 µg NPs/mL resulted in lesser viability of cells. PAEs sorbed to NPs decreased free phase concentrations (Cfree) of PAEs, which resulted in a corresponding lesser bioavailability and joint toxicity at the lesser concentration of NPs. The opposite effect was observed at the greater concentration of NPs, which may result from the dominated role of NPs in the combined toxicity. Furthermore, our data showed that oxidative stress and inflammatory reactions were mechanisms for combined cytotoxicities of PAEs and NPs on A549 cells. Results of this study emphasized the combined toxic effects and mechanisms on human lung cells, which are helpful for assessing the risk of the co-exposure of NPs and organic contaminants in humans.

Cobalt-mediated multi-functional dressings promote bacteria-infected wound healing.

Wound dressings with multiple functions are required to meet the complexity of the wound healing process. The multifunctionality often leads to an increase in the complexity and difficulty in dressing preparation. To surmount this problem, we used a facile preparation and fabrication process to fabricate a multi-functional dressing by integrating four widely accessible materials: plain gauze, sodium alginate (SA), Ca2+ and Co2+. Firstly, mixed Ca2+/Co2+ ion solutions with different concentration were applied to gauzes. After drying, SA solution was added to ionized gauze and Co2+-Ca2+/Gauze/SA (Ion-GSA) composite dressings were formed easily. In vitro results showed that all Ion-GSA dressings exhibited strong mechanical properties, uniform dispersion and sustained release of Ca2+ and Co2+, and the ability to retain moisture and absorb wound exudate. Besides the above advantages, dressings prepared with 0.25 g/L Co2+ and 4 g/L Ca2+ (Co2+0.25-Ca2+4 GSA composite dressings) exhibited the best overall effect for inducing a hypoxia-like response, and favorable cytocompatibility, hemostatic property and antibacterial activity. In vivo wound healing assays revealed that Co2+0.25-Ca2+4 GSA composite dressings inhibited bacterial growth, increased local Hypoxia-inducible factor-1α (HIF-1α), vascular endothelial growth factor (VEGF), transforming growth factor-ß1 (TGF-ß1) protein expression, and accelerated full-thickness skin wound healing in mouse bacterial-infected wound model. The quick healing wounds had improved angiogenesis, macrophages regulation, re-epithelialization and dense collagen deposition. Collectively, our results indicated that Co2+0.25-Ca2+4 GSA composite dressings promote wound healing. STATEMENT OF SIGNIFICANCE: Wound dressings with integrated functionalities are required to meet complex clinical requirements. However, there is often a trade-off between reducing preparation complexity and increasing the multifunctionality of the dressing's properties. In this study, we prepared multifunctional composite dressings by a facile preparation process using widely accessible materials. The composite dressings possessed the mechanical strength of gauze, had the effective wound exudate absorption, moisture maintenance and hemostatic property capacity of calcium alginate hydrogels, and had the hypoxia-like induction and the antimicrobial effects of Co2+. These functions all together promote bacteria-infected wound healing. Thus, we believed that the composite dressings can be widely applied in skin wound repair duo to their facile preparation method and good therapeutic effect.