Altered Membrane Permeability Research Articles

Thiazine-derived compounds in inhibiting efflux pump in Staphylococcus aureus K2068, mepA gene expression, and membrane permeability alteration

Staphylococcus aureus is a bacterium responsible for resistance to multiple drugs and the efflux system is widely studied among the resistance mechanisms developed by this species. The present study evaluates the inhibition of the MepA efflux pump by thiadiazine-derived compounds. For this purpose, thiadiazine-derived compounds (IJ-14 to IJ-20) were tested against S. aureus K2068 strains. Microdilution tests were initially conducted to assess the Minimum Inhibitory Concentration (MIC) of the compounds and their efflux pump inhibition activity. In addition, fluorimetry tests were performed using BrEt emission and tests were conducted to inhibit the expression of the mepA gene. This involved comparing the bacterial gene expression with the antibiotic alone to the gene expression after combining compounds (IJ-17 and IJ-20) with the antibiotic. Furthermore, membrane permeability assessment tests and in silico molecular docking tests were performed. It was observed that the IJ17 and IJ20 compounds exhibited direct activity against the tested strain. The IJ17 compound produced significant results in the gene inhibition tests, which was also evidenced through the membrane permeability alteration test. These findings suggest that thiadiazine-derived compounds have promising effects against one of the main resistance mechanisms, with the IJ17 compound presenting observable mechanisms of action.

Antimicrobial activity and mechanisms of carbon quantum dot decorated modified zinc oxide nanoparticles against oral pathogenic bacteria

Caries represents one of the most prevalent bacterial infectious diseases in the oral cavity. Over decades, researchers have been actively seeking nanomaterials capable of inhibiting oral pathogenic bacteria. We utilized a one-pot hydrothermal method to synthesize CQDs with multicolor luminescence and CQD-loaded ZnO composite nanomaterials. FE-SEM revealed that the rod-like structure of CQDs/ZnO becomes finer with increased CQDs content. XRD and XPS confirmed the presence of all components within the nanocomposites. We compared the antimicrobial activities of ZnO, 7.7% and 14.3% CQDs/ZnO against Streptococcus mutans, Enterococcus faecalis, Escherichia coli, and Staphylococcus aureus. The 7.7% CQDs/ZnO composite exhibited the most effective antibacterial activity. The 7.7% CQDs/ZnO composite exhibited the most effective antibacterial activity. FE-SEM and ONPG assays demonstrated that 7.7% CQDs/ZnO disrupted the membrane structure and alters membrane permeability. ESR, DPBF and ROS assayed further confirmed that 1O2 produced by 7.7% CQDs/ZnO accumulated at the cell membrane and disrupts its structure.

The potential of antifungal peptides derived from Lactiplantibacillus plantarum WYH for biocontrol of Aspergillus flavus contamination

Aspergillus flavus is a notorious fungus that contaminates food crops with toxic aflatoxins, posing a serious threat to human health and the agricultural economy. To overcome the inadequacy of traditional control methods and meet consumer preferences for natural-sources additives, there is an urgent demand for novel biocontrol agents that are safe and efficient. This study aims to investigate the antifungal properties of a novel antifungal agent derived from the biologically safe Lactiplantibacillus plantarum WYH. Firstly, antifungal peptides (AFPs) with a molecular weight of less than 3kD, exhibiting remarkable temperature stability and effectively retarding fungal growth in a dose-dependent manner specifically against A. flavus, were concentrated from the fermentation supernatant of L. plantarum WYH and were named as AFPs-WYH. Further analysis demonstrated that AFPs-WYH might exert antifungal effects through the induction of oxidative stress, disruption of mitochondrial function, alteration of membrane permeability, and cell apoptosis in A. flavus. To further validate our findings, a transcriptomics analysis was conducted on A. flavus treated with 2 and 5 mg/mL of AFPs-WYH, which elucidated the potential effect of AFPs-WYH administration on the regulation of genes involved in impairing fungal development and preventing aflatoxin biosynthesis pathways. Overall, AFPs-WYH reduced the A. flavus proliferation and affected the AFB1 biosynthesis, exhibiting a promising potential for food industry applications as a biopreservative and biocontrol agent.

Exploring therapeutic approaches against Naegleria fowleri infections through the COVID box

Naegleria fowleri, known as the brain-eating amoeba, is the pathogen that causes the primary amoebic meningoencephalitis (PAM), a severe neurodegenerative disease with a fatality rate exceeding 95%. Moreover, PAM cases commonly involved previous activities in warm freshwater bodies that allow amoebae-containing water through the nasal passages. Hence, awareness among healthcare professionals and the general public are the key to contribute to a higher and faster number of diagnoses worldwide. Current treatment options for PAM, such as amphotericin B and miltefosine, are limited by potential cytotoxic effects. In this context, the repurposing of existing compounds has emerged as a promising strategy. In this study, the evaluation of the COVID Box which contains 160 compounds demonstrated significant in vitro amoebicidal activity against two type strains of N. fowleri. From these compounds, terconazole, clemastine, ABT-239 and PD-144418 showed a higher selectivity against the parasite compared to the remaining products. In addition, programmed cell death assays were conducted with these four compounds, unveiling compatible metabolic events in treated amoebae. These compounds exhibited chromatin condensation and alterations in cell membrane permeability, indicating their potential to induce programmed cell death. Assessment of mitochondrial membrane potential disruption and a significant reduction in ATP production emphasized the impact of these compounds on the mitochondria, with the identification of increased ROS production underscoring their potential as effective treatment options. This study emphasizes the potential of the mentioned COVID Box compounds against N. fowleri, providing a path for enhanced PAM therapies.

Treatment Strategies of Colistin Resistance Acinetobacter baumannii Infections.

Acinetobacter baumannii has emerged as a pressing challenge in clinical practice, mainly due to the development of resistance to multiple antibiotics, including colistin, one of the last-resort treatments. This review highlights all the possible mechanisms of colistin resistance and the genetic basis contributing to this resistance, such as modifications to lipopolysaccharide or lipid A structures, alterations in outer membrane permeability via porins and heteroresistance. In light of this escalating threat, the review also evaluates available treatment options. The development of new antibiotics (cefiderocol, sulbactam/durlobactam) although not available everywhere, and the use of various combinations and synergistic drug combinations (including two or more of the following: a polymyxin, ampicillin/sulbactam, carbapenems, fosfomycin, tigecycline/minocycline, a rifamycin, and aminoglycosides) are discussed in the context of overcoming colistin resistance of A. baumannii infections. Although most studied combinations are polymyxin-based combinations, non-polymyxin-based combinations have been emerging as promising options. However, clinical data remain limited and continued investigation is essential to determine optimal therapeutic strategies against colistin-resistant A. baumannii.

Protease SfpB plays an important role in cell membrane stability and immune system evasion in Streptococcus agalactiae

Bacteria possess the ability to develop diverse and ingenious strategies to outwit the host immune system, and proteases are one of the many weapons employed by bacteria. This study sought to identify S. agalactiae additional serine protease and determine its role in virulence. The S. agalactiae THN0901 genome features one S8 family serine peptidase B (SfpB), acting as a secreted and externally exposed entity. A S8 family serine peptidase mutant strain (ΔsfpB) and complement strain (CΔsfpB) were generated through homologous recombination. Compared to the wild-type strain THN0901, the absorption of EtBr dyes was significantly reduced (P < 0.01) in ΔsfpB, implying an altered cell membrane permeability. In addition, the ΔsfpB strain had a significantly lower survival rate in macrophages (P < 0.01) and a 61.85 % lower adhesion ability to the EPC cells (P < 0.01) compared to THN0901. In the in vivo colonization experiment using tilapia as a model, 210 fish were selected and injected with different bacterial strains at a concentration of 3 × 106 CFU/tail. At 6, 12, 24, 48, 72 and 96 h post-injection, three fish were randomly selected from each group and their brain, liver, spleen, and kidney tissues were isolated. Subsequently, it was demonstrated that the ΔsfpB strain exhibited a markedly diminished capacity for colonization in tilapia. Additionally, the cumulative mortality of ΔsfpB in fish after intraperitoneal injection was reduced by 19.92–23.85 %. In conclusion, the findings in this study have demonstrated that the SfpB plays a significant role in S. agalactiae cell membrane stability and immune evasion. The immune evasion is fundamental for the development and transmission of invasive diseases, the serine protease SfpB may be a promising candidate for the development of antimicrobial agents to reduce the transmission of S. agalactiae.

Insight into the synergistic antibacterial mechanism of bio ZnO nanoparticles synthesized from Sophora japonica flower buds extract

Developing green nanotechnology to produce safe and powerful nano-antimicrobials has become a hot direction for addressing the challenge of antibiotic resistance. In this study, to investigate the enhancement effect of bio-capping agents on the bioactivities of plant-mediated zinc oxide nanoparticles (ZnO NPs), we designed a facile one-step green method to obtain bio-functioned ZnO NPs using flavonoids-rich extract of S. japonica flower buds (ESJ). The characterization results revealed that crystalline ZnO NPs with sizes smaller than 15 nm were obtained and the resultant bio-NP consists of ZnO core capped by ESJ-derived biomolecules. The biomolecules promoted the ZnO NPs to adhere to the bacterial cells and penetrate into cells by trapping bacteria and altering membrane permeability, which enhanced the bioactivities of the green-synthesized bio ZnO NPs and reduced its cytotoxicity. And the bio ZnO NPs showed minimum inhibitory concentrations (MIC) of 0.5 and 0.125 mg/mL toward Staphylococci aureus and Escherichia coli, respectively, and had good ⋅OH and DPPH radical scavenging activities with IC50 of 0.31 and 1.55 mg/mL, respectively. In addition, the bio ZnO NPs could maintain 80% viability of NIH3T3 cells at 2*MIC (1 mg/mL). This study provides a thought for the design of highly effective antibacterial nanomaterials, which may kill bacteria by membrane-disruption action to evade drug-resistant antimicrobial mechanisms under the synergistic actions of plant-derived biomolecules.

Molecular mechanism of action of imidazolium carbosilane dendrimers on the outer bacterial membrane – From membrane damage to permeability to antimicrobial endolysin

The outer bacterial membrane of drug-resistant bacteria is a significant barrier to many antimicrobials. Therefore, the development of new antibacterials primarily focuses on damaging the outer bacterial membrane of Gram-negative bacteria. Among many membrane-disrupting substances, the most promising are cationic dendritic systems. However, the mode of action may vary among different strains due to variations in the lipid compositions of the membrane. Here, we investigated the interaction of two types of cationic imidazolium carbosilane dendrimers: one with a single cationic group (methyl imidazolium) and the other with the same cationic group but attached to a functional group (a pendant pyridyl moiety), capable of establishing interactions with membranes through H-bonding or ion–dipole electrostatic interactions. We used different models of the outer membrane of Gram-negative bacteria – Escherichia coli, Pseudomonas aeruginosa, and Acinetobacter baumannii. Additionally, we assessed the combined effect of the dendrimers and the antibacterial endolysin on P. aeruginosa. Our results show that the mechanism of action depends on the type of dendrimer and the lipid composition of the membrane. We also demonstrate that the alteration of membrane fluidity and permeability to endolysin by the methyl imidazolium and pyridyl imidazolium dendrimers may play a more significant role in antimicrobial activity compared to membrane damage caused by positively charged dendrimers.

Antibacterial Activities and Underlying Mechanisms of the Compound SYAUP-491 against Xanthomonas oryzae pv. oryzae.

SYAUP-491 is a novel alkyl sulfonamide. In this study, in vivo and in vitro tests were performed along with a proteomic analysis to determine the effects and underlying mechanisms of the antibacterial activity of SYAUP-491 against the causative agent of bacterial leaf blight in rice. The antibacterial test results suggested that SYAUP-491 exhibited significant activities against Xanthomonas oryzae pv. oryzae (Xoo) in vitro and in vivo. The minimal EC50 values reached 6.96 μg/mL and the curative activity reached 74.1%. Detailed studies demonstrated that SYAUP-491 altered membrane permeability and caused morphological changes. Based on proteomics results, SYAUP-491 might inhibit bacterial protein synthesis. SYAUP-491 may disrupt and alter cell membrane permeability and could further act on ribosomes in the bacterial body. Given the above results, SYAUP-491 could serve as a new lead compound in the research of antibacterial control of plant pathogenic bacterial disease.

Synergistic Antibacterial Activity of Curcuma domestica Val. Extract with Tetracycline Against Multidrug-resistant Acinetobacter baumannii

Infections caused by multidrug-resistant (MDR) bacteria are on the rise globally. MDR is facilitated by overexpression of efflux pump and permeability changes of membrane. Acinetobacter baumannii is a pathogenic germ that causes a major problem in infection, also, there has been an increase in the incidence of resistance to various antibiotics. The present study highlights the synergistic of ethanolic extract of Curcuma domestica (EECD) rhizome with tetracycline against multidrug-resistant A. baumannii (MDR-Ab). Assessment of Minimum Inhibitory Concentration (MIC) was determined by microdilution using 96-well plates. The synergistic effect of EECD and tetracycline was determined by checkerboard method. The effect of EECD and tetracycline combination was investigated by bacteriolytic activity and inhibition of efflux pump by Ethidium Bromide (EtBr) accumulation assay. EECD presented the MIC value 250 µg/mL against MDR-Ab. Fractional Inhibitory Concentration Index (FICI) value of EECD and tetracycline combination was 0.4, which showed their synergistic effect. Additionally, the combination of EECD and tetracycline could inhibit the efflux pump in MDR-Ab. This combination can also compromise cell integrity by altering membrane permeability thus lysing the bacteria cells. According to these results, EECD and tetracycline combination has synergistic effects at some sites of action, and thus could be used as a breakthrough to overcome infection problems due to MDR-Ab.&#x0D; &#x0D; Keywords: Acinetobacter baumannii, antibacterial, Curcuma domestica, multidrug-resistant, tetracycline

Polystyrene nanoparticles induce biofilm formation in Pseudomonas aeruginosa

In recent years, micro/nanoplastics have garnered widespread attention due to their ecological risks. In this study, we investigated the effects of polystyrene nanoparticles (PS-NPs) of different sizes on the growth and biofilm formation of Pseudomonas aeruginosa PAO1. The results demonstrated that exposure to certain concentrations of PS-NPs significantly promoted bacterial biofilm formation. Meanwhile, we comprehensively revealed its mechanism whereby PS-NPs induced oxidative stress and altered bacterial membrane permeability by contacting or penetrating bacterial membranes. To counteract the stimulation by PS-NPs and reduce their toxicity, bacteria enhanced biofilm formation by upregulating the expression of biofilm-related genes, increasing EPS and virulence factors secretion, and enhancing bacterial motility through the participation of the quorum sensing (QS) system. Additionally, we also found that exposure to PS-NPs enhanced bacterial antibiotic resistance, posing a challenge to antimicrobial therapy. Our study reveals the toxic effects of nanoplastics and the defense mechanisms of bacteria, which has important implications for the risk assessment and management of environmental nanoplastics.

Smilax aspera L. Leaf and Fruit Extracts as Antibacterial Agents for Crop Protection

Smilax aspera L. (commonly known as sarsaparilla) is recognized for its composition rich in flavonoids, phenylpropanoids, steroidal saponins, stilbenoids, and tannins, exhibiting anti-inflammatory, cytotoxic, and antimicrobial properties. This study investigates the hydromethanolic extracts of its leaves and fruits through vibrational spectroscopy and gas chromatography–mass spectrometry, evaluating their potential as biorationals for safeguarding crops. Analysis of S. aspera leaf and fruit extracts revealed the presence of phytochemicals such as lactones and other furan derivatives. In vitro assessments against three phytopathogens—Erwinia amylovora, Pseudomonas syringae pv. actinidiae, and Xanthomonas campestris pv. campestris—demonstrated strong antibacterial activity, with minimum inhibitory concentration (MIC) values of 1500 μg·mL−1 for both extracts. Biofilm tests indicated that the leaf extract reduced biofilm formation by 78–85%, while the fruit extract led to a reduction of 73–92.5%. At a concentration of 750 µg·mL−1, the extracts caused a decrease in amylovoran synthesis by 41–58%. Additionally, noticeable alterations in membrane permeability were observed at MIC and MIC/2 doses. Subsequent in vivo trials conducted on Pyrus communis L. trees utilizing the combined aerial part extract yielded substantial protection against E. amylovora at a dose of 1500 μg·mL−1, reaching 80% wilting reduction for the leaf extract. The findings presented herein cast S. aspera extracts as a promising natural-based treatment against these bacterial phytopathogens.

A translocation fluorescent probe for analyzing cellular physiological parameters in neurological disease models.

Neurological disorders are closely linked to the alterations in cell membrane permeability (CMP) and mitochondrial membrane potential (MMP). Changes in CMP and MMP may lead to damage and death of nerve cells, thus triggering the onset and progression of neurological diseases. Therefore, monitoring the changes of these two physiological parameters not only benefits the accurate assessment of nerve cell health status, but also enables providing key information for the diagnosis and treatment of neurological diseases. However, the simultaneous monitoring of these two cellular physiological parameters is still challenging. Herein, we design and synthesize two quinolinium-carbazole-derivated fluorescent probes (OQ and PQ). As isomers, the only difference in their chemical structures is the linking position of the carbazole unit in quinoline rings. Strikingly, such a subtle difference endows OQ and PQ with significantly different organelle-staining behaviors. PQ mainly targets at the nucleus, OQ can simultaneously stain cell membranes and mitochondria in normal cells, and performs CMP and MMP-dependent translocation from the cell membrane to mitochondria then to the nucleus, thus holding great promise as an intracellular translocation probe to image the changes of CMP and MMP. After unraveling the intrinsic mechanism of their different translocation abilities by combining experiments with molecular dynamics simulations and density functional theory calculations, we successfully used OQ to monitor the continuous changes of CMP and MMP in three neurological disease-related cell models, including oxidative stress-damaged, Parkinson's disease, and virus-infected ones. Besides providing a validated imaging tool for monitoring cellular physiological parameters, this work paves a promising route for designing intracellular translocation probes to analyze cellular physiological parameters associated with various diseases.

A membrane targeted multifunctional cationic nanoparticle conjugated fusogenic nanoemulsion (CFusoN): induced membrane depolarization and lipid solubilization to accelerate the killing of Staphylococcus aureus.

Bacterial infections caused by Staphylococcus aureus are one of the growing concerns for human health care management globally. Antibiotic-associated adverse effects and the emergence of bacterial resistant strains necessitate the development of an alternative yet effective approach. Nanoemulsion-based therapy has emerged as a potential therapeutic strategy to combat bacterial infestation. Herein, we designed a cationic metal nanoparticle-conjugated fusogenic nanoemulsion (CFusoN) as a lipid solubilizing nanovesicle for the effective treatment of S. aureus infection with a killing efficiency of 99.999%. The cationic nanoparticle-conjugated nanoemulsion (viz. NECNP) (24.4 ± 2.9 mV) electrostatically bound with the negatively charged bacterial cell membrane (-10.2 ± 3.7 mV) causing alteration of the bacterial surface charge. The fluorometric and flow cytometry studies confirmed the bacterial membrane depolarization and altered cell membrane permeability leading to cell death. The atomic force microscopic studies further demonstrated the damage of the cellular ultrastructure, while the transmission electron microscopic image and membrane lipid solubilization analysis depicted the solubilization of the bacterial membrane lipid bilayer along with the leakage of the intracellular contents. The cell membrane fatty acid analysis revealed that the methyl esters of palmitic acid, stearic acid and octadecadienoic acid isomers were solubilized after the treatment of S. aureus with CFusoN. The bactericidal killing efficiency of CFusoN is proposed to occur through the synergistic efficacy of the targeted attachment of CNP to the bacterial cells along with the lipid solubilization property of NE. Interestingly, NECNP didn't elicit any in vitro hemolytic activity or cytotoxicity against red blood cells (RBCs) and L929 fibroblast cells, respectively, at its bactericidal concentration. Furthermore, a porcine skin wound infection model exhibited the enhanced wound cleansing potency of CFusoN in comparison to the commercially available wound cleansers. The obtained antibacterial activity, biocompatibility and skin wound disinfection efficacy of the NECNP demonstrated the formulation of a cell targeted CFusoN as a promising translatable strategy to combat bacterial infection.

Synthesis and antibacterial activity of magnetic Fe3O4-loaded silver nanocomposites

The problem of drug-resistant bacteria arising from antibiotic misuse is a growing global health concern. Therefore, there is an urgent need to develop effective antimicrobial agents. In this study, Fe3O4 nanospheres with superparamagnetic properties were first prepared by the "solvothermal method" and then coated with dopamine. The coordination principle was then utilized for the adsorption of ultra-small Ag nanocomposites (Ag NPs) on the surface of Fe3O4 @PDA nanocomposite (FPN). The material morphology and crystallinity of the prepared composites were characterized using TEM, XRD, XPS, UV–vis, and FT-IR analysis techniques. The performance and mechanism of inhibition of Gram-negative Escherichia coli (E. coli), Gram-positive Bacillus subtilis (B. subtilis), and drug-resistant Salmonella typhi (S. typhi) by Fe3O4 @PDA@Ag nanocomposite (FAN) were also studied. The results showed that FAN with an inhibitory concentration of 200 μg/mL achieved 99.9% inhibition against E. coli, B. subtilis, and S. typhi at 60 min. Zeta potential, PI staining, ion leakage, bacterial growth curves, cytoplasmic leakage, and biocompatibility experiments were used further to explore the probable bacterial inhibition mechanisms of the composites. The results of the experiments done to determine the inhibition mechanism suggest that the composite material disrupts the negative environment of the bacterial cell wall, altering membrane permeability and ultimately leading to rupture and death. The findings of this study demonstrate that FAN possess excellent antibacterial properties and have a promising future in wastewater treatment and biomedical applications.

Regulation of the Gene for Alanine Racemase Modulates Amino Acid Metabolism with Consequent Alterations in Cell Wall Properties and Adhesive Capability in Brucella spp.

Brucella, a zoonotic facultative intracellular pathogenic bacterium, poses a significant threat both to human health and to the development of the livestock industry. Alanine racemase (Alr), the enzyme responsible for alanine racemization, plays a pivotal role in regulating virulence in this bacterium. Moreover, Brucella mutants with alr gene deletions (Δalr) exhibit potential as vaccine candidates. However, the mechanisms that underlie the detrimental effects of alr knockouts on Brucella pathogenicity remain elusive. Here, initially, we conducted a bioinformatics analysis of Alr, which demonstrated a high degree of conservation of the protein within Brucella spp. Subsequent metabolomics studies unveiled alterations in amino acid pathways following deletion of the alr gene. Furthermore, alr deletion in Brucella suis S2 induced decreased resistance to stress, antibiotics, and other factors. Transmission electron microscopy of simulated macrophage intracellular infection revealed damage to the cell wall in the Δalr strain, whereas propidium iodide staining and alkaline phosphatase and lactate dehydrogenase assays demonstrated alterations in cell membrane permeability. Changes in cell wall properties were revealed by measurements of cell surface hydrophobicity and zeta potential. Finally, the diminished adhesion capacity of the Δalr strain was shown by immunofluorescence and bacterial enumeration assays. In summary, our findings indicate that the alr gene that regulates amino acid metabolism in Brucella influences the properties of the cell wall, which modulates bacterial adherence capability. This study is the first demonstration that Alr impacts virulence by modulating bacterial metabolism, thereby providing novel insights into the pathogenic mechanisms of Brucella spp.

Sub-lethal concentrations of chlorhexidine inhibit Candida albicans growth by disrupting ROS and metal ion homeostasis

ABSTRACT Candida albicans is a normal resident of the human oral cavity. It is also the most common fungal pathogen, causing various oral diseases, particularly in immunocompromised individuals. Chlorhexidine digluconate (CHG) is a broad-spectrum antimicrobial agent widely used in dental practice and has been recommended to treat oral candidiasis. However, its action mechanism against the fungal pathogen C. albicans remains poorly understood. The aim of the present study was to investigate the effect of CHG at sub-lethal concentrations against C. albicans. CHG inhibited the growth of C. albicans in a dose- and time-dependent manner. Cells treated with CHG exhibited altered membrane permeability, reduced metabolic activity, and enhanced metal ion and reactive oxygen species (ROS) accumulation. Copper-sensing transcription factor Mac1, iron-sensing transcription factors Sfu1 and Sef2, and copper transporter Ctr1 regulated intracellular metal ion and ROS homeostasis in response to CHG. Deletion of MAC1, SFU1, or SEF2 increased intracellular ROS production and cell susceptibility to CHG. This study revealed a novel mechanism by which CHG induced apoptosis of C. albicans cells through the disruption of metal ion and ROS homeostasis, which may help to identify new targets for fungal infections.

The efficient activity of plant essential oils for inhibiting Botrytis cinerea and Penicillium expansum: Mechanistic insights into antifungal activity

Botrytis cinerea and Penicillium expansum produce deterioration in fruit quality, causing losses to the food industry. Thus, plant essential oils (EOs) have been proposed as a sustainable alternative for minimizing the application of synthetic fungicides due to their broad-spectrum antifungal properties. This study investigated the efficacy of five EOs in suppressing the growth of B. cinerea and P. expansum and their potential antifungal mechanisms. EOs of Mentha × piperita L., Origanum vulgare L., Thymus vulgaris L., Eucalyptus globules Labill., and Lavandula angustifolia Mill., were screened for both fungi. The results showed that the EO of T. vulgaris and O. vulgare were the most efficient in inhibiting the growth of B. cinerea and P. expansum. The concentration increase of all EO tested increased fungi growth inhibition. Exposure of fungi to EOs of T. vulgaris and O. vulgare increased the pH and the release of constituents absorbing 260 nm and soluble proteins, reflecting membrane permeability alterations. Fluorescence microscopic examination revealed that tested EOs produce structural alteration in cell wall component deposition, decreasing the hypha width. Moreover, propidium iodide and Calcein-AM stains evidenced the loss of membrane integrity and reduced cell viability of fungi treated with EOs. Fungi treated with EOs decreased the mitochondria activity and the respiratory process. Therefore, these EOs are effective antifungal agents against B. cinerea and P. expansum, which is attributed to changes in the cell wall structure, the breakdown of the cell membrane, and the alteration of the mitochondrial activity.

In vitro and in vivo anti-oomycetes activities and mechanisms of linalool against Saprolegnia ferax

Saprolegnia ferax (S. ferax) is one of the most highly pathogenic oomycetes species in freshwater aquatic environments causing serious saprolegniosis. Linalool is a kind of natural alternative of antibiotics existing in various plants essential oils. In this study, the in vitro and in vivo anti-oomycetes activity and mode of action of linalool against S. ferax CQT1 were investigated. For in vitro test, linalool exhibited mycelium growth and zoospore production inhibition at 0.1% (v/v, 0.862 mg/ml) and 0.025% (v/v, 0.216 mg/ml), respectively. The mycelium was wrinkled, deformed, and shrunken after treatment with linalool. Linalool altered membrane permeability, caused permanent damage, or membrane breakdown. Further, the transcriptomic analysis showed that: (1) Linalool affected the oxidation and metabolism of fatty acids, leading to mitochondrial dysfunction and the inhibition of energy supply. (2) The increase in glycerophospholipid metabolism is probably the cell response to deal with cellular damage. (3) The degradation of branched chain amino acids may inhibit amino acid metabolism and result in the dysfunction of protein synthesis. (4) ABC transporters contribute to metabolic resistance to linalool and efflux of linalool outside of cells. For in vivo test, the cumulative survival rate of grass carp treated with linalool was significantly higher than those untreated with linalool, moreover, microscopic histopathology analysis results showed that there was robust mucus cell density in fish skin, the dermis and epidermis were relatively intact after linalool treatment. In all, our findings would provide foundations for the potential use of linalool to prevent and control S. ferax infection in the aquaculture industry.

The blood-prostate barrier: an obstacle to drug delivery into the prostate.

The blood-prostate barrier (BPB), a non-static physical barrier, stands as an obstacle between the prostate stroma and the lumen of the prostate gland tube. The barrier has the ability to dynamically regulate and strictly control the mass exchange between the blood and the prostate, thereby limiting drug penetration into the prostate. The basement membrane, fibrous stromal layer, capillary endothelial cell, prostatic ductal epithelial cell, lipid layer, etc., have been confirmed to be involved in the composition of the barrier structure and altered membrane permeability mainly by regulating the size of paracellular ion pores. Various studies have been conducted to improve the efficiency of drug therapy for prostate diseases by changing the administration approaches, improving barrier permeability and increasing drug penetration. To gain a full understanding of BPB, the composition of BPB, the methodology for evaluating the permeability of BPB and alterations in barrier function under pathological conditions are summarized in this review. To find a shortcut for drug delivery across BPB, the biodistribution of drugs in the prostate and different methods of improving drug penetration across BPB are outlined. This review offers an applied perspective on recent advances in drug delivery across BPB.