Cell Membrane Permeability Research Articles

The effects of small plastic particles on antibiotic resistance gene transfer revealed by single cell and community level analysis

Small plastic particles with sizes comparable to bacterial cells, widely exist in environment. However, their effects on antibiotic resistance gene (ARG) dissemination remain unclear. Using polystyrene (PS) particles (0.2 µm, 2 µm, 5 µm, 10 µm, 15 µm, 20 µm) as models, conjugative transfer of ARGs between the donor E. coli and different recipients (E. coli or sludge bacterial community) was investigated. Compared to the pure strain, the sludge bacterial community exposed to PS particles showed higher transfer frequencies (1.67 to 14.31 times the blank control). The transfer frequencies first decreased and then increased with particle size, and plastics similar in size to bacteria (e.g., 2 µm) appear to be a transitional zone with minimal impact on ARG transmission. Furthermore, using microfluidics, in-situ observation at single cell level found that 2 µm plastics can act as barriers between donor and recipient bacteria inhibiting growth, but conjugation events mostly occurred around them. Conversely, nanoplastics (e.g., 0.2 µm) and larger microplastics (e.g., 20 µm) significantly promote conjugation, mainly due to increased reactive oxygen species production and cell membrane permeability, or facilitating bacterial adhesion and biofilm formation, respectively. This study aids in assessing environmental risks of small plastic particles on ARG dissemination.

Antifungal Effect of Oregano Essential Oil Against Penicillium expansum on Pyrus sinkiangensis.

Given the increasing demand for fruit safety assurance and environmental protection, plant essential oils have gained significant attention as natural alternatives for control of postharvest decay caused by various pathogens. In postharvest management, it is particularly important to effectively control postharvest decay without compromising the quality attributes of fruits. Although oregano essential oil (OEO) has been extensively studied for its antimicrobial properties against various postharvest pathogens, few studies have focused on its interactions with postharvest fruits. In this study, OEO was applied for management of postharvest decay of Pyrus sinkiangensis caused by Penicillium expansum, and its antifungal mechanisms and impacts on the quality attributes of pears were investigated. The OEO exhibited notable inhibitory effects, with determined MIC (0.02%) and MFC (0.04%) against P. expansum, which highlighted its potential as a viable alternative to synthetic fungicides. Our findings revealed that OEO disrupted P. expansum by compromising the integrity of the fungal plasma membrane, as evidenced by changes in plasma membrane permeability and the leakage of cellular components. The OEO treatment significantly reduced weight loss, maintained firmness, and preserved soluble-solid content in the treated pears. In addition, OEO treatment stimulated the intrinsic antioxidant mechanisms of pears, as demonstrated by elevated activities of superoxide dismutase and catalase during storage. This study provides compelling evidence for the antifungal and quality-preserving properties of OEO in the postharvest management of pears, underscoring its potential as an alternative to synthetic fungicides for controlling postharvest decay. The elucidation of the interaction between OEO and P. sinkiangensis would greatly enhance our comprehensive understanding of the biological activities of OEO and facilitate its practical application in the postharvest management of pears.

Effect of High CO2 Controlled Atmosphere Storage on Postharvest Quality of Button Mushroom (Agaricus bisporus).

The Agaricus bisporus (Button mushroom) stands out as one of the most prolific edible fungi which offers robust flavor and nutrition. Nonetheless, this mushroom contains high moisture levels and intense respiration. Without appropriate postharvest preservation techniques, the button mushroom readily experiences browning and senescence. To ensure optimum quality, prompt cooling and appropriate storage conditions are essential. This present research investigated the postharvest quality of button mushrooms stored in a controlled atmosphere (CA) with different initial gas compositions. The findings revealed that button mushrooms in the CA group demonstrated considerable enhancements in appearance and overall quality, effectively delaying browning and senescence compared to those in the control group. The optimal gas composition is 1-3% O2 and 15-17% CO2 (CAII), which effectively inhibited the expression of polyphenol oxidase (PPO)- and lactase (LAC)-related genes in the button mushroom, maintaining a high L* value. Furthermore, the application of 1-3% O2 and 15-17% CO2 (CAII) not only preserved visual quality but also extended the postharvest shelf life of the button mushroom by minimizing metabolic activities that contribute to senescence. Moreover, 1-3% O2 and 15-17% CO2 (CAII) storage also reduced the expression levels of genes associated with ethylene synthesis, which is reflected in the gradual decrease in cell membrane permeability. Consequently, this research underscores the critical importance of controlled atmosphere storage in improving the marketability and sustainability of this widely consumed mushroom.

Meta-analysis of SnRK2 gene overexpression in response to drought and salt stress.

SNF1-RELATED KINASE 2 (SnRK2) plays a crucial role in plants' stress response. Although studies have reported that the overexpression of several SnRK2 family members in different plants leads to improved stress tolerance, it is difficult to elucidate the mechanisms by which SnRK2s regulate stress tolerance due to the variability of experimental variables in these studies. Therefore, we used meta-analysis to comprehensively analyze 22 parameters that can reflect drought tolerance and salinity tolerance in SnRK2s-transformed plants and to explore the effects that different experimental variables between studies have on the relevant plant parameters. The results showed that the overexpression of SnRK2s mainly improved plants' drought and salinity tolerance by reducing their osmotic stress and oxidative damage, improving photosynthesis and other biochemical and physiological processes. Out of the 22 physiological parameters, 17 and 19 were significantly affected by drought and salt stress, respectively, and 10 indicators were also significantly changed under non-stress conditions. Under salt stress, the cell membrane permeability among these parameters shows the most significant changes, increasing by 506.57% in SnRK2-overexpressing plants compared to wild type (WT). Therefore, although plants overexpressing SnRK2s respond positively to both drought and salt stress, they demonstrated greater tolerance to salt stress. In addition, among the detected regulatory variables, donor-acceptor type, promoter type, stress type, experimental medium, and duration all affected the extent of SnRK2s overexpression and affected the physiological characteristics of the transgenic plants. Also, different stress conditions (salt, drought stress) led to different degrees of transformation. These studies provide new research directions for studying crop stress tolerance and help to better explore the functions played by SnRK2s in external plant stresses.

Cuminaldehyde in combination with tetracycline shows promising antibiofilm activity against drug-resistant Pseudomonas aeruginosa

Pseudomonas aeruginosa, an opportunistic pathogen often causes biofilm-linked infections. A combinatorial approach involving tetracycline (antibiotic) and cuminaldehyde (phytochemical) was explored to combat this infectious pathogen. The results showed that both tetracycline and cuminaldehyde individually demonstrated antibacterial effects. However, when the compounds were applied together, there was a significant increase in their antimicrobial potential. The determined fractional inhibitory concentration index of 0.43 indicated a synergistic interaction between the two compounds. Furthermore, a series of experiments demonstrated that the combined application of cuminaldehyde and tetracycline could lead to a significant enhancement of their antibiofilm potential. This enhanced antibiofilm potential was attributed to the accumulation of reactive oxygen species and increased cell membrane permeability. Besides, this combinatorial application reduced the secretion of various virulence factors from P. aeruginosa. Therefore, this combined approach holds promise for effectively treating P. aeruginosa biofilms.

Performance and potential mechanisms for reactive electrochemical ceramic membrane system to inhibit resistance transmission in antibiotic-resistant contaminated wastewater: From a microbial perspective

Due to the widespread presence of antibiotic resistance genes (ARGs) in traditional biological wastewater treatment systems, electrochemical disinfection techniques have received extensive attention. The rTNA/Sb-SnO2/PbO2 Reactive Electrochemical Ceramic Membrane (RECM) system has demonstrated outstanding performance in effectively removing refractory antibiotics. However, its capacity to remove antibiotic resistant bacteria (ARB) remains unclear. In this study, an antibiotic resistant strain of Escherichia coli (AR E. coli) was employed to investigate the disinfection efficiency of the RECM system at a macroscopic level, along with its disinfection mechanism at a microscopic level. Furthermore, the secondary effluent from an antibiotic production wastewater treatment plant was collected to validate the disinfection performance of this system against ARB in actual wastewater. In the RECM system, the inactivation of AR E. coli and the removal of ARGs were primarily governed by the electrocatalytic process, with notable influences from critical factors such as current density, electrolyte type, and hydraulic retention time. Morphological observation and flow cytometry experiments confirmed that the AR E. coli in the RECM system was inactivated by alterations in cell morphology, leading to fragmentation, and by improvement in cell membrane permeability. More significantly, this system exhibited noteworthy efficiency in treating wastewater contaminated with antibiotic resistant pollutants, effectively eliminating both antibiotics and ARGs from the secondary effluent. Overall, our study comprehensively investigated the performance and underlying mechanisms of the RECM system in mitigating the dissemination of ARGs, as well as its potential feasibility of practical application.

MG53 Protein-Mediated biomimetic nanotherapeutics for the treatment of acute myocardial infarction by driving the impaired plasma membrane resealing

Myocardial infarction (MI) remains a leading global cause of mortality. The frequent disruptions of cardiomyocyte plasma membranes underscore the pivotal role played by MG53, a tissue repair protein secreted by skeletal muscle in response to physical exercise, in cardioprotection. However, the clinical application of recombinant human MG53 (rhMG53) is challenged by its short circulation half-life, necessitating frequent administration, and its low cell membrane permeability. Here we introduce an innovative MG53 protein-mediated biomimetic nanotherapeutic (Rm@mSiO2@MG53) to address the poor delivery. Rm@mSiO2@MG53 nanotherapeutics, attributed to the high specific surface area and adjustable pore size of mSiO2 and the expression of CD47 on erythrocyte membranes, enable sustained protein delivery, enhance their ability to evade the mononuclear phagocytic system (MPS), and extend the half-life of rhMG53 proteins in circulation. In a mouse MI model, Rm@mSiO2@MG53 demonstrated superior efficacy in protecting injured cardiomyocytes and preserving heart function post-MI. Our findings developed a novel protein biomimetic nanotherapeutics that mediates heart repair by driving cell membrane resealing, offering the potential to expedite the clinical deployment of rhMG53 as a viable therapeutic for treating MI and other tissue injuries.

Supramolecular Self-Assembled Hydrogel for Antiviral Therapy through Glycyrrhizic Acid-Enhanced Zinc Absorption and Intracellular Accumulation.

Respiratory syncytial virus (RSV) is a common pathogen that causes respiratory infections in infants and children worldwide, significantly impacting hospitalization rates in this age group. Zinc ions are considered to have broad-spectrum antiviral potential against RNA viruses, including RSV. However, poor organism absorption and low intracellular accumulation of zinc require repeated high-dose supplementation, which may lead to unnecessary toxic side effects. In this research, a Zn2+-mediated glycyrrhizinic acid (GA)-based hydrogel (ZnGA Gel) was introduced and potentially developed to be a clinically available drug candidate for RSV therapy. ZnGA Gel was fabricated based on the cooperation of two potential RSV inhibiting molecules (Zn2+ and GA), where Zn2+ promoted self-assembly of GA and reduced its gel concentration and GA promoted zinc absorption and distribution in lung tissue in vivo. The facile construction of supramolecular hydrogel by the self-assembled coordination complex made it an injectable, temperature-sensitive, and pH-responsive controlled-release drug delivery for Zn2+. Most importantly, GA was observed to enhance organism absorption and intracellular accumulation of Zn2+ and was identified as a zinc ionophore for the first time. GA can colonize on the cell membrane and disturb cell membrane potential, resulting in an enhanced cell membrane permeability. In the presence of GA, more than 4.7-fold increasing Zn2+ concentrations materialized in the intracellular cytoplasm, compared to Zn2+ alone administration. This intracellular Zn2+ accumulation directly boosted the antiviral activities through improved inhibition of RSV replication-associated proteins and significantly inhibited RSV replication. Oral administration of ZnGA Gel on the RSV-infected mice model achieved an ideal therapeutic effect by effectively lowering viral load in the lungs, alleviating lung injury symptoms, and reducing inflammatory cell infiltration at pathological sites. The mechanism involved the inhibition of RSV replication-related proteins, aligning with our in vitro results. Additionally, ZnGA Gel had demonstrated biocompatibility, and reasonable supplementation of zinc was acceptable and effective for infants and children in clinical practice. Hence, the ZnGA Gel developed by us holds promise as an effective anti-RSV medicine in the future.

Design, synthesis and antifungal activity of arylhydrazine analogs containing diphenyl ether fragments.

Succinate dehydrogenase (SDH) represents a critical target in the development of novel fungicides. To address the growing issue of resistance and safeguard the economic viability of agricultural production, the pursuit of new succinate dehydrogenase inhibitors (SDHIs) has emerged as a significant focus of contemporary research. In this project, 32 arylhydrazine derivatives containing diphenyl ether structural units were synthesized and evaluated for their fungicidal activities against Rhizoctonia solani, Sclerotinia sclerotiorum, Alternaria alternata, Gibberella zeae, Alternaria solani and Colletotrichum gloeosporioides. In an in vitro fungicidal activity assay, compound D6 showed significant inhibitory activity against R. solani with a half-maximum effective concentration (EC50) of 0.09 mg L-1. The in vivo fungicidal activity demonstrated that compound D6 inhibited R. solani by 95.39% in rice leaves, which was significantly better than that of boscalid (85.76%). The results of SDH enzyme assay, molecular docking simulation, mitochondrial membrane potential assay, cytoplasmic release studies and morphological observations demonstrated that the target compound D6 not only had significant SDH inhibitory activity, but also affected the membrane integrity of mycelium. Bioactivity screening and validation of the mechanism of action indicated that compound D6 was a potentially unique SDHI, acting on SDH while also affecting cell membrane permeability, which deserved further study. © 2024 Society of Chemical Industry.

A respiratory Streptococcus strain inhibits Acinetobacter baumannii from causing inflammatory damage through ferroptosis

BackgroundMicroecological equilibrium is essential for human health. Previous research has demonstrated that Streptococcus strain A, the main bacterial group in the respiratory tract, can suppress harmful microbes and protect the body. In this study, Streptococcus strain D19T was isolated from the oral and pharyngeal cavities of healthy children. Its antibacterial mechanism against Acinetobacter baumannii was examined, as well as its potential to prevent inflammatory damage to cells. We evaluated the effect of the fermentation conditions of D19T on inhibition of Acinetobacter baumannii growth; Isolation and purification of antibacterial active components of strain D19T and molecular mechanism of inhibition of Acinetobacter baumannii; Molecular mechanism of D19T antibacterial protein reversing cellular inflammatory injury induced by Acinetobacter baumannii.ResultsThe supernatant of fermentation broth of Streptococcus D19T was the active component against Acinetobacter baumannii, but the bacteria had no antibacterial activity. The supernatant of D19T fermentation broth was precipitated by (NH4)2SO4 solution, and the protein was the active antibacterial component. After gel filtration chromatography and anion gel filtration chromatography, the molecular weight of antibacterial protein was 53kD. D19T antibacterial protein can improve cell membrane permeability, limit extracellular soluble protein release, inhibit Acinetobacter baumannii biofilm formation, and prevent Acinetobacter baumannii adhesion. Acinetobacter baumannii induces inflammatory damage to respiratory cells via ferroptosis, and the D19T antibacterial protein can counteract this damage, protecting the respiratory tract.ConclusionStreptococcus strain D19T, as a potential probiotic, inhibits the growth of Acinetobacter baumannii and the inflammatory damage of respiratory cells, playing a protective role in human respiratory health.

Efficient electroporation in primary cells with PEDOT:PSS electrodes.

Precise and efficient delivery of macromolecules into cells enhances basic biology research and therapeutic applications in cell therapies, drug delivery, and personalized medicine. While pulsed electric field electroporation effectively permeabilizes cell membranes to deliver payloads without the need for toxic chemical or viral transduction agents, conventional bulk electroporation devices face major challenges with cell viability and heterogeneity due to variations in fields generated across cells and electrochemistry at the electrode-electrolyte interface. Here, we introduce the use of microfabricated electrodes based on the conducting polymer poly(3,4-ethylenedioxythiophene) doped with polystyrene sulfonate (PEDOT:PSS), which substantially increases cell viability and transfection efficiency. As a proof of concept, we demonstrate the enhanced delivery of Cas9 protein, guide RNA, and plasmid DNA into cell lines and primary cells. This use of PEDOT:PSS enables rapid modification of difficult-to-transfect cell types to accelerate their study and use as therapeutic platforms.

Baseline Sensitivity and Toxicity Mechanisms of Prochloraz to Alternaria alternata Strains Associated with Maize Leaf Blight in Heilongjiang Province in China.

Alternaria species are fungal pathogens that can infect maize, causing leaf blight disease and significant economic losses. This study aimed to determine the baseline sensitivity to prochloraz of A. alternata isolates obtained from diseased maize leaves collected from Heilongjiang Province by assessing the half-maximal effective concentration (EC50) values. The EC50 values of prochloraz ranged from 0.0550 to 2.3258 μg/ml, with an average of 0.9995 ± 0.5192 μg/ml. At EC50 (1.2495 μg/ml) and 2EC50 (2.4990 μg/ml), prochloraz increased the number of mycelial offshoots, disrupted the cell membrane integrity of conidia and mycelia, and resulted in a reduced ergosterol content in the mycelia. Prochloraz significantly affected the mycelial cell membrane permeability and increased the malondialdehyde content and superoxide dismutase activity. No cross-resistance was detected between prochloraz and other fungicides. These data demonstrate that prochloraz is a promising fungicide for managing maize leaf blight caused by A. alternata and provide novel insights into understanding the mechanism of prochloraz toxicity against A. alternata isolates.

Lipid bilayer permeabilities and antibiotic effects of tetramethylguanidinium and choline fatty acid ionic liquids

Ionic liquids (ILs) have been studied as potential components in antibiotic formulations based on their abilities to permeabilize and penetrate lipid bilayer, which correlate with their antibacterial effects. Fatty acid-based ILs (FAILs), in which the anion is a long-chain fatty acid, can permeabilize lipid membranes and have been used in biomedical applications since they have low human cell cytotoxicity. In this work we investigated the abilities of several different FAILs to permeabilize lipid bilayers and how that permeabilization correlates with antibacterial activity, cell membrane permeability, and cytotoxicity. The FAILs consisted of the cations tetramethylguanidinium (TMG) or choline combined with octanoate or decanoate. These FAILs were tested on model bilayer vesicles with three different lipid compositions for membrane permeabilization using a leakage assay. They were then tested for antibiotic and membrane permeabilization on bacterial and mammalian cells. The results show that while the octanoate-based FAILs do not form micelles and have low activities on vesicles and biological cells, the decanoate-based FAILs can permeabilize bilayers and have biological activities that correlate with the model vesicle results. The ILs with both cation and fatty-acid anion have strong activities while the decanoate alone has only minimal permeabilization and antibiotic activity. Membrane permeabilization occurs at FAIL concentrations below their CMC values which suggests that their mechanism of action may not involve micelle formation.

Immunomodulation of human T cells by microbubble-mediated focused ultrasound.

While met with initial and ground-breaking success targeting blood borne cancers, cellular immunotherapy remains significantly hindered in the context of solid tumors by the tumor microenvironment. Focused ultrasound, in conjunction with microbubbles, has found tremendous potential as a targeted and local drug/gene delivery technique for cancer therapy. The specific immunomodulating effects of this technique on immune cells, including T-cells, remain unexplored. Here, with freshly isolated human immune cells, we examine how focused ultrasound can viably modulate immune cell membrane permeability and influence the secretion of over 90 cytokines, chemokines and other analytes relevant to a potent immune response against cancer. We determine that microbubble-mediated focused ultrasound modulates the immune cell secretome in a time-dependent manner - ranging in ~0.1-3.6-fold changes in the concentration of a given cytokine compared to sham controls over 48 hours post-treatment (e.g. IL-1β, TNF-α, CX3CL1, CCL21). Further, we determine the general trend of a negative correlation between secreted cytokine concentration and viable ultrasound-assisted membrane permeability with negligible loss of cell viability. Taken together, the data presented here highlights the potential of microbubble-mediated focused ultrasound to viably enhance T-cell permeability and modulate key pro-immune pathways, offering a novel approach to augment targeted cellular therapies for solid tumors.

Effect of low-frequency alternating magnetic field on exopolysaccharide production and antioxidant capacity of Pleurotus citrinopileatus by submerged fermentation.

The objective of this study was to investigate the effect of low-frequency alternating magnetic field (LF-AMF) on the production of extracellular polysaccharide (EPS) by submerged fermentation of Pleurotus citrinopileatus. The fermentation conditions optimized by the central composite design method were as follows: fermentation time of 6.18days, temperature of 28.28°C, shaking speed of 149.04 r/min, and inoculum amount of 8.43%. Under these conditions, a LF-AMF was applied to the submerged fermentation of P. citrinopileatus. When the intensity of LF-AMF was 40 Gs, the initial intervention time was 24h after inoculation, and the treatment time was 6h at one time, the mycelial biomass of P. citrinopileatus increased by 11.30%, and the EPS yield increased by 23.09% compared with the fermentation without LF-AMF treatment. The morphology of mycelium after LF-AMF treatment was observed by scanning electron microscopy. It was found that the surface of mycelium was wrinkled, and the structure of mycelium was loose, which might be more conducive to the production of EPS. Mycelium diameter decreased, and ATPase activity increased, indicating that LF-AMF had a positive effect on the production of EPS by P. citrinopileatus fermentation. Moreover, LF-AMF could improve the permeability of the mycelial cell membrane, facilitate the exchange of intracellular and extracellular substances, and increase the metabolic capacity of P. citrinopileatus. In vitro antioxidant test of EPS showed that LF-AMF treatment also improved its antioxidant capacity.

Novel Flavonol Derivatives Containing Quinoxaline: Insights into the Antifungal Mechanism against Sclerotinia sclerotiorum.

In this study, 12 pairs of tautomeric flavonol derivatives containing quinoxaline were synthesized. The results of antifungal activity showed that in the enol-keto tautomerism, the target compounds containing keto (YB series) had better inhibitory activity against Sclerotinia sclerotiorum (S.s.) than compounds containing enol (YA series). YB9 showed the strongest antifungal activity against S.s., and the median effective concentration (EC50) value was 1.0 μg/mL, which was better than azoxystrobin (Az, 35.3 μg/mL). In vivo fungal inhibition experiments showed that the protective activity of YB9 against rape leaves was 83.4% at 200 μg/mL, which was superior to that of Az (70.2%). The activity of succinate dehydrogenase and molecular docking results showed that YB9 had a stronger antifungal effect than YA9. The results of oxalic acid content determination showed that YB9 could reduce the pathogenic ability of S.s. Then, the inhibitory effect of YB9 against S.s. was further verified by scanning electron microscopy, fluorescence microscopy, cell membrane permeability, cell content leakage, and malondialdehyde content.

Effects of Clausena lansium leaves volatile oil emulsion against Staphylococcus aureus in mice via autophagy modulation

BackgroundVolatile oil from fresh Clausena lansium (Lour.) Skeels (Rutaceae) (common name Wampee) has been previously extracted by our group from fresh C. lansium leaf and its components were qualitative and quantitatively analyzed by GC–MS. It altered the cell membrane permeability of Staphylococcus aureus and reduced the levels of inflammation factors. However, previous in vivo reports on the anti-inflammatory and the antibacterial properties against S. aureus are scarce. Hypothesis/PurposeTo evaluate the protective in vivo effects of Wampee leaves volatile oil emulsion (WVOE) against S. aureus-induced pneumonia and elucidate the underlying mechanisms of action. MethodsWild-type and nucleotide oligomerization domain-like receptor protein 3 (NLRP3)-deficient mice were used. Mice were treated with WVOE for 7 days, and subjected to S. aureus infection by nasal administration on day 5 for 48 h. Lung and blood samples were collected for assessing lung damage and protein abundance. Lung bacterial load, wet/dry ratio, C-reactive protein (CRP) levels, inflammatory cytokines secretion, and lung histopathological injury were examined. ResultsWVOE effectively reduced lung bacterial load, wet/dry ratio, and CRP levels increased following S. aureus infection in mice. WVOE decreased the secretion of inflammatory cytokines (IL-6 and TNF-α) and lung histopathological injury, and suppressed the NF-κB pathway and NLRP3 inflammasome activation. NLRP3-/- mice exhibited lower bacterial load, inflammatory cytokines levels and lung histopathological injury compared with mice in the model group. Autophagy was enhanced in S. aureus-infected mice, with higher levels of p-mTOR, Beclin-1, Atg 16L1, Atg7, p62, p-p62, and LC3II. WVOE administration restored the autophagy related protein levels. Autophagy was inhibited in NLRP3-/- mice of the control and model groups, and WVOE lost its ability to regulate the autophagy-related proteins enhanced upon S. aureus infection. WVOE enhanced autophagy to alleviate lung injury by inhibiting NLRP3-targeted P62. Furthermore, compared with the 3MA + model group, WVOE reduced the bacterial load and CRP levels, pulmonary septa narrowing, and congestion. NLRP3 protein expression increased due to autophagy inhibition. WVOE exerted a pharmacological effect through the PI3K/AKT/mTOR pathway. ConclusionWVOE regulated the PI3K/AKT/mTOR pathway and enhanced autophagy, with NLRP3 playing a crucial role. WVOE exhibited protective effects against S. aureus-induced pneumonia by inhibiting NLRP3 inflammasome activation and enhancing autophagy. These findings expand the understanding of antibacterial properties of WVOE, and provide novel insights into the therapeutic potential of WVOE in managing S. aureus infections.

Unlocking Intracellular Protein Delivery by Harnessing Polymersomes Synthesized at Microliter Volumes using Photo-PISA.

Efficient delivery of therapeutic proteins and vaccine antigens to intracellular targets is challenging due to generally poor cell membrane permeation and endolysosomal entrapment causing degradation. Herein, these challenges are addressed by developing an oxygen-tolerant photoinitiated polymerization-induced self-assembly (Photo-PISA) process, allowing for the microliter-scale (10µL) synthesis of protein-loaded polymersomes directly in 1536-well plates. High-resolution techniques capable of analysis at a single particle level are employed to analyze protein encapsulation and release mechanisms. Using confocal microscopy and super-resolution stochastic optical reconstruction microscopy (STORM) imaging, their ability to deliver proteins into the cytosol following endosomal escape is subsequently visualized. Lastly, the adaptability of these polymersomes is exploited to encapsulate and deliver a prototype vaccine antigen, demonstrating its ability to activate antigen-presenting cells and support antigen cross-presentation for applications in subunit vaccines and cancer immunotherapy. This combination of ultralow volume synthesis and efficient intracellular delivery holds significant promise for unlocking the high throughput screening of a broad range of otherwise cost-prohibitive or early-stage therapeutic protein and vaccine antigen candidates that can be difficult to obtain in large quantities. The versatility of this platform for rapid screening of intracellular protein delivery can result in significant advancements across the fields of nanomedicine and biomedical engineering.

Precision Delivery of Binary Cooperative Nanodrugs Self-Assembled by Berberine Glycyrrhetinic Acid Salts for Hepatocellular Carcinoma Treatment.

Berberine (BB) has demonstrated significant inhibitory effects on tumorigenesis and progression. However, its clinical application is hindered by challenges such as low bioavailability due to limited cell membrane permeability, nontargeted accumulation away from solid tumors, and increased toxicity associated with intravenous administration. To overcome these challenges, a lipophilic salt-forming strategy was employed to enhance lipophilicity, thereby promoting membrane permeability and targetability for tumor accumulation while simultaneously mitigating the toxicity associated with intravenous injection. In vitro findings revealed an almost 10-fold increase in fluorescence intensity with BB-GA NDs compared to BB alone. Furthermore, selective cytotoxicity against tumor cells exhibited a 4-fold elevation compared to normal cells. In vivo results underscored the remarkable tumor-selective accumulation of BB-GA NDs, effectively mitigating the intravenous injection toxicity associated with pure BB. The self-assembly of binary cooperative nanodrugs utilizing berberine glycyrrhetinic acid salts opens up innovative possibilities for drug delivery systems in traditional Chinese medicine.

Unveiling the roles of CaSDH8 in Candida albicans: Implications for virulence and azole resistance

ABSTRACT Candida albicans is the most common pathogen in systemic fungal diseases, exhibits a complex pathogenic mechanism, and is increasingly becoming drug tolerant. Therefore, it is particularly important to study the genes associated with virulence and resistance of C. albicans. Here, we identified a gene (orf19.1588) that encodes a conserved mitochondrial protein known as CaSDH8, upon deletion of CaSdh8, the deleted strain (Casdh8Δ/Δ) experienced impaired growth, hyphal development, and virulence. Casdh8Δ/Δ displayed a reduced capacity to utilize alternative carbon sources, along with detrimental alterations in reactive oxygen species (ROS), mitochondrial membrane potential (MMP) depolarization, and adenosine triphosphate (ATP) levels. Interestingly, Casdh8Δ/Δ demonstrated resistance to azole drugs, and under the influence of fluconazole, the cell membrane permeability and mitochondrial function of Casdh8Δ/Δ were less compromised than those of the wild type, indicating a reduction in the detrimental effects of fluconazole on Casdh8Δ/Δ. These findings highlight the significance of CaSDH8 as a crucial gene for the maintenance of cellular homoeostasis. Our study is the first to document the effects of the CaSDH8 gene on the virulence and azole resistance of C. albicans at both the molecular and animal levels, providing new clues and directions for the antifungal infection and the discovery of antifungal drug targets.