Bacterial Membrane Permeability Research Articles

Constituents, Antibacterial Effect, and Cytotoxicity of Essential Oil from Aerial Parts of Notopterygium incisum.

Notopterygium incisum Ting ex H. T. Chang (N. incisum) is a precious Chinese traditional medicine distributed in high-altitude regions of southwest China. The aim of this study was to investigate the composition, antibacterial activity, and cytotoxicity of essential oil from aerial parts of N. incisum. N. incisum essential oil (NI-EO) was extracted by hydro-distillation, and gas chromatography/mass spectrometry (GC-MS) analysis showed that the major components of NI-EO were D-limonene (18.42%) and γ-terpinene (15.03%). The antibacterial activity and mechanism study showed that the diameters of inhibition zone (DIZs) of NI-EO against E. coli and S. aureus were 14.63 and 11.25mm and the minimum inhibitory concentrations were 3.75 and 7.5μL/mL, respectively. NI-EO not only caused intracellular biomacromolecule leakage and cell deformation by destroying bacterial cell wall integrity and cell membrane permeability, but also degraded the mature biofilm. The low toxicity of NI-EO was demonstrated in an assay on bovine mammary epithelial cells. These results implied that NI-EO was mainly composed of monoterpenes and sesquiterpenes and had excellent antibacterial activity and showed low levels of cytotoxicity. It is expected to be applied as a natural antibacterial agent in the future.

Mechanistic Assessment of Metabolic Interaction between Single Oral Commensal Cells by Scanning Electrochemical Microscopy.

The human oral microbiome heavily influences the status of oral and systemic diseases through different microbial compositions and complex signaling between microbes. Recent evidence suggests that investigation of interactions between oral microbes can be utilized to understand how stable communities are maintained and how they may preserve health. Herein, we investigate two highly abundant species in the human supragingival plaque, Streptococcus mitis and Corynebacterium matruchotii, to elucidate their real-time chemical communication in commensal harmony. Specifically, we apply nanoscale scanning electrochemical microscopy (SECM) using a submicropipet-supported interface between two immiscible electrolyte solutions as an SECM probe not only to image the permeability of S. mitis and C. matruchotii membranes to tetraethylammonium (TEA+) probe ions but also to real-time visualize the metabolic interaction between two microbes via lactate production/consumption at a single-cell level. The metabolic relationship between two strains is quantitatively assessed by determining (1) the passive permeability of both bacterial membranes of 2.4 × 10-4 cm/s to the free diffusion of TEA+, (2) 0.5 mM of the lactate concentration produced by a single S. mitis strain at a rate of 2.7 × 10-4 cm/s, and (3) a lactate oxidation rate ≥5.0 × 106 s-1 by an individual C. matruchotii strain. Significantly, this study, for the first time, describes a mechanism of in situ metabolic interaction between oral commensals at the single-cell level through quantitative analysis, which supports the observed in vivo spatial arrangements of these microbes.

Antibacterial Activity and Mechanisms of TroHepc2-22, a Derived Peptide of Hepcidin2 from Golden Pompano (Trachinotus ovatus).

Hepcidin, a cysteine-rich antimicrobial peptide, has a highly conserved gene structure in teleosts, and it plays an essential role in host immune response against various pathogenic bacteria. Nonetheless, few studies on the antibacterial mechanism of hepcidin in golden pompano (Trachinotus ovatus) have been reported. In this study, we synthesized a derived peptide, TroHepc2-22, from the mature peptide of T. ovatus hepcidin2. Our results showed that TroHepc2-22 has superior antibacterial abilities against both Gram-negative (Vibrio harveyi and Edwardsiella piscicida) and Gram-positive (Staphylococcus aureus and Streptococcus agalactiae) bacteria. Based on the results of a bacterial membrane depolarization assay and propidium iodide (PI) staining assay in vitro, TroHepc2-22 displayed antimicrobial activity by inducing the bacterial membrane depolarization and changing the bacterial membrane permeability. Scanning electron microscopy (SEM) visualization illustrated that TroHepc2-22 brought about membrane rupturing and the leakage of the cytoplasm for the bacteria. In addition, TroHepc2-22 was verified to have hydrolytic activity on bacterial genomic DNA in view of the results of the gel retardation assay. In terms of the in vivo assay, the bacterial loads of V. harveyi in the tested immune tissues (liver, spleen, and head kidney) were significantly reduced in T. ovatus, revealing that TroHepc2-22 significantly enhanced the resistance against V. harveyi infection. Furthermore, the expressions of immune-related genes, including tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), interleukin 1-β (IL-1β), IL-6, Toll-like receptor 1 (TLR1), and myeloid differentiation factor 88 (MyD88) were significantly increased, indicating that TroHepc2-22 might regulate inflammatory cytokines and activate immune-related signaling pathways. To summarize, TroHepc2-22 possesses appreciable antimicrobial activity and plays a vital role in resisting bacterial infection. The observation of our present study unveils the excellent application prospect of hepcidin as a substitute for antibiotics to resist pathogenic microorganisms in teleosts.

Functional annotation of rhizospheric phageome of the wild plant species Moringa oleifera.

The study aims to describe phageome of soil rhizosphere of M.oleifera in terms of the genes encoding CAZymes and other KEGG enzymes. Genes of the rhizospheric virome of the wild plant species Moringa oleifera were investigated for their ability to encode useful CAZymes and other KEGG (Kyoto Encyclopedia of Genes and Genomes) enzymes and to resist antibiotic resistance genes (ARGs) in the soil. Abundance of these genes was higher in the rhizospheric microbiome than in the bulk soil. Detected viral families include the plant viral family Potyviridae as well as the tailed bacteriophages of class Caudoviricetes that are mainly associated with bacterial genera Pseudomonas, Streptomyces and Mycobacterium. Viral CAZymes in this soil mainly belong to glycoside hydrolase (GH) families GH43 and GH23. Some of these CAZymes participate in a KEGG pathway with actions included debranching and degradation of hemicellulose. Other actions include biosynthesizing biopolymer of the bacterial cell wall and the layered cell wall structure of peptidoglycan. Other CAZymes promote plant physiological activities such as cell-cell recognition, embryogenesis and programmed cell death (PCD). Enzymes of other pathways help reduce the level of soil H2O2 and participate in the biosynthesis of glycine, malate, isoprenoids, as well as isoprene that protects plant from heat stress. Other enzymes act in promoting both the permeability of bacterial peroxisome membrane and carbon fixation in plants. Some enzymes participate in a balanced supply of dNTPs, successful DNA replication and mismatch repair during bacterial cell division. They also catalyze the release of signal peptides from bacterial membrane prolipoproteins. Phages with the most highly abundant antibiotic resistance genes (ARGs) transduce species of bacterial genera Pseudomonas, Streptomyces, and Mycobacterium. Abundant mechanisms of antibiotic resistance in the rhizosphere include "antibiotic efflux pump" for ARGs soxR, OleC, and MuxB, "antibiotic target alteration" for parY mutant, and "antibiotic inactivation" for arr-1. These ARGs can act synergistically to inhibit several antibiotics including tetracycline, penam, cephalosporin, rifamycins, aminocoumarin, and oleandomycin. The study highlighted the issue of horizontal transfer of ARGs to clinical isolates and human gut microbiome.

CuCeO Bimetallic Oxide Rapidly Treats Staphylococcus aureus-Infected Osteomyelitis through Microwave Strengthened Microwave Catalysis and Fenton-Therapy.

Osteomyelitis caused by bacteria is a deep-seated lesion and is often treated clinically with antibiotics. Long-term use of antibiotics may predispose bacteria to develop resistance. Here, CuCeOx material is applied to treat infectious bacterial osteomyelitis using microwave (MW)-assisted bacterial killing. Heat generation occurs as a result of the dielectric properties of the material under MW irradiation, and the material generates reactive oxygen species (ROS) under MW irradiation. Heat and ROS increase the thermal sensitivity and permeability of bacterial cell membranes, and the released copper ions easily penetrate the bacterial membrane and react with H2 O2 to produce a toxic hydroxyl group inside the bacteria, leading to the bacteria's eventual death. This is due to the synergistic effect of the MW thermal effect, ROS, and the breaking of the equilibrium within the bacteria. CuCeOx can effectively treat osteomyelitis caused by Staphylococcus aureususing MW irradiation. This study can safely and effectively address the challenge of deep tissue infections by shedding light on non-invasive antimicrobial systems and using MW thermal therapy and MW dynamics to achieve therapeutic results.

Antibacterial Activity and Mechanism of Peptide PV-Q5 against Vibrio parahaemolyticus and Escherichia coli, Derived from Salt-Fermented Penaeus vannamei.

The increasing threat posed by antibiotic-resistant pathogens has prompted a shift to the use of naturally-derived antimicrobial peptides (AMPs) in place of chemical preservatives in controlling foodborne pathogens. In this study, ten peptides were identified from salt-fermented shrimps (Penaeus vannamei) using ultra-performance liquid chromatography-mass spectrometry. One of the peptides, designated PV-Q5 (QVRNFPRGSAASPSALASPR), with most features of an AMP, was further explored and found to possess strong antibacterial activity against Vibrio parahaemolyticus and Escherichia coli, with a minimum inhibitory concentration of 31.25 μg/mL. Moreover, PV-Q5 increased bacterial cell membrane permeability and ruptured bacteria cell membranes, as revealed by transmission electron microscopy. Circular dichroism analysis showed that the conformation of PV-Q5 was a random coil in phosphate-buffered saline and α-helical in sodium dodecyl sulfate, which is conducive for interaction with bacteria cell membranes. These findings indicated that PV-Q5 could find potential use in food preservation to control foodborne pathogenic bacteria.

Transition between Different Diffusion Modes of Individual Lipids during the Membrane-Specific Action of As-CATH4 Peptides.

The cell membrane permeabilization ability of immune defense antimicrobial peptides (AMPs) is widely applied in biomedicine. Although the mechanisms of peptide-membrane interactions have been widely investigated, analyses at the molecular level are still lacking. Herein, the membrane-specific action of a native AMP, As-CATH4, is investigated using a single-lipid tracking method in combination with live cell and model membrane assays conducted at different scales. The peptide-membrane interaction process is characterized by analyzing single-lipid diffusion behaviors. As-CATH4 exhibits potent antimicrobial activity through bacterial membrane permeabilization, with moderate cytotoxicity against mammalian cells. In-plane diffusion analyses of individual lipids show that the lipid molecules exhibit non-Gaussian and heterogeneous diffusion behaviors in both pristine and peptide-treated membranes, which can be decomposed into two Gaussian subgroups corresponding to normal- and slow-diffusive lipids. Assessment of the temporal evolution of these two diffusion modes of lipids reveal that the peptide action states of As-CATH4 include surface binding, transmembrane defect formation, and dynamic equilibrium. The action mechanisms of As-CATH4 at varying concentrations and against different membranes are distinguished. This work resolves the simultaneous mixed diffusion mechanisms of single lipids in biomimetic cell membranes, especially during dynamic membrane permeabilization by AMPs.

Na+/K+ carrier ionophore antibiotics valinomycin and monensin enhance the antibacterial activity of fluoride.

Fluoride is routinely used as a highly effective antibacterial agent that interferes with bacterial metabolism through fundamentally different mechanisms. One of the major bacterial evasion mechanisms against fluoride is the impermeability of cell envelope to the anion that limits its cellular uptake. Therefore, translating such compounds to clinical settings requires novel mechanisms to facilitate the uptake of membrane-impermeant molecules. Published data have indicated antibiotic synergy between fluoride and membrane destabilizing agents that induce strong fluoride toxicity in bacteria via enhancing the permeability of bacterial membranes to fluoride. Here, we report a similar mechanism of antibiotic synergy between fluoride and potassium ion carriers, valinomycin and monensin against Gram-positive bacteria, B. subtilis and S. aureus. Molecular dynamics simulations were performed to understand the effect of potassium on the binding affinity of fluoride to monensin and valinomycin. The trajectory results strongly indicated that the monensin molecules transport fluoride ions across the cell membrane via formation of ion-pair between the monensin-K+ complex and a fluoride. This study provides new insights to design novel compounds to enhance the uptake of small toxic anions via synergistic interactions and thus exert strong antibacterial activity against a wide variety of pathogens.

Thymol-Decorated Gold Nanoparticles for Curing Clinical Infections Caused by Bacteria Resistant to Last-Resort Antibiotics.

Multidrug-resistant bacteria pose a tremendous challenge to public health worldwide. Many bacteria resistant to last-resort antibiotics due to antibiotic misuse have been recently reported, which may give rise to serious infections without effective treatment. Therefore, it is imperative to develop novel antimicrobial strategies. Natural phenols are known to increase bacterial membrane permeability and are potential candidates for the development of new antimicrobial agents. In this study, gold nanoparticles (Au NPs) carrying natural phenols were synthesized to combat bacteria resistant to last-resort antibiotics. Transmission electron microscopy, dynamic light scattering, zeta potential, and UV-visible spectra were used to characterize the synthesized Au NPs, which showed good monodispersity and uniform particle size. Evaluation of antibacterial activity using the broth microdilution method revealed that thymol-decorated gold nanoparticles (Thymol_Au NPs) had a broad antibacterial spectrum and higher bactericidal effects than last-resort antibiotics against last-resort-antibiotic-resistant bacteria. Considering the underlying antibacterial mechanism, the results showed that Thymol_Au NPs destroyed bacterial cell membranes. Further, Thymol_Au NPs were effective in treating mouse abdominal infections and exhibited acceptable biocompatibility without any significant toxicity in cell viability and histopathological assays, respectively, at most bactericidal concentrations. However, attention should be paid to changes in white blood cells, reticulocyte percentages, and superoxide dismutase activity during Thymol_Au NP treatment. In conclusion, Thymol_Au NPs have the potential for treating clinical infections caused by bacteria resistant to last-resort antibiotics. IMPORTANCE Excessive use of antibiotics can lead to bacterial resistance and the development of multidrug-resistant bacteria. Antibiotic misuse can also promote resistance against last-resort antibiotics. It is thus crucial to develop alternatives to antibiotics to retard the development of multidrug resistance. In recent years, the use of several nanodosage forms of antibacterial drugs has been investigated. These agents kill bacteria through a variety of mechanisms and avoid the problem of resistance. Among them, Au NPs, which are safer to use for medical applications than other metal nanoparticles, have attracted interest as potential antibacterial agents. To combat bacterial resistance to last-resort antibiotics and mitigate the problem of antimicrobial resistance, it is important and meaningful to develop antimicrobial agents based on Au NPs.

Berberine disrupts staphylococcal proton motive force to cause potent anti-staphylococcal effects in vitro

The presence of antibiotic resistance has increased the urgency for more effective treatments of bacterial infections. Biofilm formation has complicated this issue as biofilm bacteria become tolerant to antibiotics due to environmental factors such as nutrient deprivation and adhesion. In septic arthritis, a disease with an 11% mortality rate, bacteria in synovial fluid organize into floating, protein-rich, bacterial aggregates (mm-cm) that display depressed metabolism and antibiotic tolerance. In this study, Staphylococcus aureus (S. aureus), which is the most common pathogen in septic arthritis, was tested against different inhibitors that modulate bacterial surface protein availability and that should decrease bacterial aggregation. One of these, berberine, a quaternary ammonium compound, was found to reduce bacterial counts by 3–7 logs in human synovial fluid (aggregating medium) with no effect in tryptic soy broth (TSB, non-aggregating). Unlike traditional antibiotics, the bactericidal activity of berberine appeared to be independent of bacterial metabolism. To elucidate the mechanism, we used synovial fluid fractionation, targeted MRSA transposon insertion mutants, dyes to assess changes in membrane potential (DiSC3(5)) and membrane permeability (propidium iodide (PI)), colony counting, and fluorescence spectroscopy. We showed that berberine's activity was dependent on an alkaline pH and berberine killed both methicillin-sensitive S. aureus and MRSA in alkaline media (pH 8.5–9.0; p < 0.0001 vs. same pH controls). Under these alkaline conditions, berberine localized to S. aureus where berberine was isolated in cytoplasmic (∼95%) and DNA (∼5%) fractions. Importantly, berberine increased bacterial cell membrane permeability, and disrupted the proton motive force, suggesting a mechanism whereby it may be able to synergize with other antibacterial compounds under less harsh conditions. We suggest that berberine, which is cheap and readily available, can be made into an effective treatment.

Short, Lipidated Dendrimeric γ-AApeptides as New Antimicrobial Peptidomimetics.

Antibiotic resistance is one of the most significant issues encountered in global health. There is an urgent demand for the development of a new generation of antibiotic agents combating the emergence of drug resistance. In this article, we reported the design of lipidated dendrimeric γ-AApeptides as a new class of antimicrobial agents. These AApeptides showed excellent potency and broad-spectrum activity against both Gram-positive bacteria and Gram-negative bacteria, including methicillin-resistant Staphylococcus aureus (MRSA). The mechanistic studies revealed that the dendrimeric AApeptides could kill bacteria rapidly through the permeabilization of bacterial membranes, analogous to host-defense peptides (HDPs). These dendrimers also did not induce antibiotic resistance readily. The easy access to the synthesis, together with their potent and broad-spectrum activity, make these lipidated dendrimeric γ-AApeptides a new generation of antibacterial agents.

Smart Trap‐Capture‐Kill Antibacterial System for Infected Microenvironment Improvement and Vascularized Bone Regeneration via Magnetic Thermotherapy

AbstractInfected bone defect has gradually become a disastrous complication in orthopedic surgery because of the bacterial biofilm formation both on the surface of the implants and surrounding tissues, which will cause antibiotic resistance and weak postoperative osseointegration. Herein, drug‐loaded hollow mesoporous ferrite nanoparticles coated by gelatin (GM/HMFNs) with pomegranate structure are prepared and compounded into calcium magnesium phosphate cement (MCPC) as a Trap‐Capture‐Kill system to kill bacteria and inhibit biofilm formation efficiently. Under the action of alternating magnetic field (AMF), MCPC/GM/HMFNs/Drug composites exhibit acute magnetic hyperthermal capacity and recruit the free bacteria. Then, the permeability of bacterial cell membrane is increased via reactive oxygen species (ROS) damage of nano‐MgO and HMFNs, providing channels through which vancomycin (Van) and ciprofloxacin (CPFX) encapsulated in GM/HMFNs can penetrate. Subsequently, the bacteria are efficiently killed under the synergetic cooperation of magnetothermal effect, ROS damage, and adsorption damage of nano‐MgO, and the bioactive ions and drugs. The antibacterial system also presents outstanding pro‐osteogenesis and pro‐angiogenic capacity in vitro. And it promotes the vascularized bone regeneration of infective femoral defect in SD rats in vivo, providing promising potential for the treatment of infected bone defects through magnetothermal therapy.

Water-Soluble Dicationic Deuteroporphyrin Derivative for Antimicrobial PDT: Singlet Oxygen Generation, Passive Carrier Interaction and Nosocomial Bacterial Strains Photoinactivation

Multidrug resistance of pathogenic microflora is a serious threat to the modern community looking for new approaches to treating superinfections. In this sense, antimicrobial photodynamic therapy (aPDT) is an effective and safe technique considered to be a promising alternative or an important supplement to the traditional clinically applied methods for inactivating antibiotic resistant pathogens. Macroheterocyclic photosensitizers (PS) of three generations are proposed for clinical practice. They are known as the key compounds for PDT able to be localized selectively in microbial cells and to be activated with the red light producing toxic reactive oxygen species (ROS). However, these neutral and anionic PSs possess low affinity towards the outer lipopolysaccharide membrane of Gram-negative bacteria and, consequently, poor ability to kill these pathogens under irradiation. In contrast, cationic PSs containing one or more charged groups, especially those bound to an appropriate carrier, provide efficient inactivation of microorganisms. In this paper, we focus on the study of photophysics, aggregation and photoinduced antimicrobial activity of the water-soluble derivative of deuteroporphyrin-IX, a blood group porphyrin, bearing two cationic trialkylammonium fragments. This potential photosensitizing agent is found to generate singlet oxygen in a non-polar environment and forms stable nano-sized molecular complexes with passive non-ionic carrier Tween 80, localizing in an aqueous surfactant solution as a non-aggregated form in the surface micellar layer. Two different modes of PS/Tween 80 binding characterized by their own stability constants and interaction stoichiometry are observed. Microbiological experiments clearly demonstrate that the increased permeability of the outer bacterial membrane caused by the application of the intramicellar form of the photosensitizer or addition of some potentiation agents leads to pronounced light phototoxicity of the pigment against antibiotic-resistant nosocomial strains of Gram-negative bacterial pathogens.

Antimicrobial Peptide Cec4 Eradicates Multidrug-Resistant Acinetobacter baumannii in vitro and in vivo.

Acinetobacter baumannii has become a major difficulty in the treatment of bacteria-associated infection. The previously reported antimicrobial peptide Cec4 exhibited good and stable activity against A. baumannii in vitro, but the mechanisms and effects in vivo are elusive. The effects of Cec4 on bacterial membrane permeability, membrane potential and bacterial reactive oxygen species were measured. The cell membrane localization of antimicrobial peptides was studied by fluorescence labelling. The ability of bacteria to develop resistance to antimicrobial peptides was studied by continuous induction, and transcriptome difference was analysed. The in vivo toxicity of Cec4 against nematodes and mice was studied, and the in vivo therapeutic potential of Cec4 against A. baumannii was assessed. Cec4 effectively cleared multidrug-resistant A. baumannii by altering bacterial cell membrane permeability, changing bacterial cell membrane polarity, and increasing bacterial intracellular reactive oxygen species. Cec4 affected the expression of the secretion system, outer membrane, and efflux pump genes of A. baumannii. In addition, the bacteria did not acquire stable drug-resistant ability. Cec4 at 1.024 mg/mL did not affect the proliferation of HeLa and HepG2 cells, and Cec4 at 45 mg/kg had little effect on the mortality of Caenorhabditis elegans, even the liver and kidney tissues of mouse. Most importantly, Cec4 could effectively improve the survival rates and reduce the bacterial load of various tissues in the mouse model of infection. In conclusion, Cec4 can damage the cell membrane of bacteria, and the bacteria is not easy to produce resistance to Cec4. Besides, Cec4 has good potential for the treatment of multidrug-resistant A. baumannii infections.

Comparative study on antibacterial characteristics of the multiple liver expressed antimicrobial peptides (LEAPs) in teleost fish.

Antimicrobial peptides are important components of the host innate immune system, forming the first line of defense against infectious microorganisms. Among them, liver-expressed antimicrobial peptides (LEAPs) are a family of antimicrobial peptides that widely exist in vertebrates. LEAPs include two types, named LEAP-1 and LEAP-2, and many teleost fish have two or more LEAP-2s. In this study, LEAP-2C from rainbow trout and grass carp were discovered, both of which are composed of 3 exons and 2 introns. The antibacterial functions of the multiple LEAPs were systematically compared in rainbow trout and grass carp. The gene expression pattern revealed that rainbow trout and grass carp LEAP-1, LEAP-2A, LEAP-2B and/or LEAP-2C were differentially expressed in various tissues/organs, mainly in liver. After bacterial infection, the expression levels of LEAP-1, LEAP-2A, LEAP-2B and/or LEAP-2C in the liver and gut of rainbow trout and grass carp increased to varying degrees. Moreover, the antibacterial assay and bacterial membrane permeability assay showed that rainbow trout and grass carp LEAP-1, LEAP-2A, LEAP-2B and LEAP-2C all have antibacterial activities against a variety of Gram-positive and Gram-negative bacteria with varying levels through membrane rupture. Furthermore, cell transfection assay showed that only rainbow trout LEAP-1, but not LEAP-2, can lead to the internalization of ferroportin, the only iron exporter on cell surface, indicating that only LEAP-1 possess iron metabolism regulation activity in teleost fish. Taken together, this study systematically compared the antibacterial function of LEAPs in teleost fish and the results suggest that multiple LEAPs can enhance the immunity of teleost fish through different expression patterns and different antibacterial activities to various bacteria.

Antibacterial Components and Modes of the Methanol-Phase Extract from Commelina communis Linn.

Infectious diseases caused by pathogenic bacteria severely threaten human health. Traditional Chinese herbs are potential sources of new or alternative medicine. In this study, we analyzed for the first time antibacterial substances in the methanol-phase extract from a traditional Chinese herb-Commelina communis Linn-which showed an inhibition rate of 58.33% against 24 species of common pathogenic bacteria. The extract was further purified using preparative high-performance liquid chromatography (Prep-HPLC), which generated four single fragments (Fragments 1 to 4). The results revealed that Fragment 1 significantly increased bacterial cell surface hydrophobicity and membrane permeability and decreased membrane fluidity, showing disruptive effects on cell integrity of Gram-positive and Gram-negative bacteria, such as Bacillus cereus, Enterococcus faecalis, Staphylococcus aureus, and Salmonella enterica subsp., compared to the control groups (p < 0.05). In sum, 65 compounds with known functions in Fragment 1 were identified using liquid chromatography and mass spectrometry (LC-MS), of which quercetin-3-o-glucuronide was predominant (19.35%). Comparative transcriptomic analysis revealed multiple altered metabolic pathways mediated by Fragment 1, such as inhibited ABC transporters, ribosome, citrate cycle and oxidative phosphorylation, and upregulated nitrogen metabolism and purine metabolism, thereby resulting in the repressed bacterial growth and even death (p < 0.05). Overall, the results of this study demonstrate that Fragment 1 from C. communis Linn is a promising candidate against common pathogenic bacteria.

Study of Antibacterial Properties of Cinnamaldehyde against Aeromonas hydrophila

Antibacterial properties of cinnamaldehyde against Aeromonas hydrophila were assayed in this study. To investigate the action mechanisms of cinnamaldehyde against A. hydrophila, we examined the antibacterial activity, bacterial membrane permeability, and ultrastructure of A. hydrophila cells treated with cinnamaldehyde. The results showed that the minimum inhibitory concentration (MIC) value of cinnamaldehyde against A. hydrophila NJ-35 was found to be 0.039 mg/mL. The trends of the growth curve of A. hydrophila treated with different concentrations of cinnamaldehyde (from 1/4 MIC to 2 MIC) were different, and there was a significant difference in the growth curve of different groups of treatment. There were significant differences in the K+ concentration among all treatment groups from 1 h to 5 h after incubation compared with that of the control. The highest K+ concentration was observed in the 1 MIC group of cinnamaldehyde. The ultrastructure of A. hydrophila cells treated with cinnamaldehyde was destroyed, and the morphology changed. These results indicated that cinnamaldehyde could inhibit the growth of A. hydrophila, increase bacterial membrane permeability, and damage cell membrane integrity, resulting in leakage of the A. hydrophila cell contents.

Alkaline "Nanoswords" Coordinate Ferroptosis-like Bacterial Death for Antibiosis and Osseointegration.

Ferroptosis is an iron-dependent cell death and is associated with cancer therapy. Can it play a role in resistance of postoperative infection of implants, especially with an extracellular supplement of Fe ions in a non-cytotoxic dose? To answer this, "nanoswords" of Fe-doped titanite are fabricated on a Ti implant surface to resist bacterial invasion by a synergistic action of ferroptosis-like bacteria killing, proton disturbance, and physical puncture. The related antibiosis mechanism is explored by atomic force microscopy and genome sequencing. The nanoswords induce an increased local pH value, which not only weakens the proton motive force, reducing adenosine triphosphate synthesis of Staphylococcus aureus, but also decreases the membrane modulus, making the nanoswords distort and even puncture a bacterial membrane easily. Simultaneously, more Fe ions are taken by bacteria due to increased bacterial membrane permeability, resulting in ferroptosis-like death of bacteria, and this is demonstrated by intracellular iron enrichment, lipid peroxidation, and glutathione depletion. Interestingly, a microenvironment constructed by these nanoswords improves osteoblast behavior in vitro and bone regeneration in vivo. Overall, the nanoswords can induce ferroptosis-like bacterial death without cytotoxicity and have great promise in applications with clinical implants for outstanding antibiosis and biointegration performance.

Gossypol acetate: A natural polyphenol derivative with antimicrobial activities against the essential cell division protein FtsZ.

Antimicrobial resistance has attracted worldwide attention and remains an urgent issue to resolve. Discovery of novel compounds is regarded as one way to circumvent the development of resistance and increase the available treatment options. Gossypol is a natural polyphenolic aldehyde, and it has attracted increasing attention as a possible antibacterial drug. In this paper, we studied the antimicrobial properties (minimum inhibitory concentrations) of gossypol acetate against both Gram-positive and Gram-negative bacteria strains and dig up targets of gossypol acetate using in vitro assays, including studying its effects on functions (GTPase activity and polymerization) of Filamenting temperature sensitive mutant Z (FtsZ) and its interactions with FtsZ using isothermal titration calorimetry (ITC), and in vivo assays, including visualization of cell morphologies and proteins localizations using a microscope. Lastly, Bacterial membrane permeability changes were studied, and the cytotoxicity of gossypol acetate was determined. We also estimated the interactions of gossypol acetate with the promising target. We found that gossypol acetate can inhibit the growth of Gram-positive bacteria such as the model organism Bacillus subtilis and the pathogen Staphylococcus aureus [both methicillin-sensitive (MSSA) and methicillin-resistant (MRSA)]. In addition, gossypol acetate can also inhibit the growth of Gram-negative bacteria when the outer membrane is permeabilized by Polymyxin B nonapeptide (PMBN). Using a cell biological approach, we show that gossypol acetate affects cell division in bacteria by interfering with the assembly of the cell division FtsZ ring. Biochemical analysis shows that the GTPase activity of FtsZ was inhibited and polymerization of FtsZ was enhanced in vitro, consistent with the block to cell division in the bacteria tested. The binding mode of gossypol acetate in FtsZ was modeled using molecular docking and provides an understanding of the compound mode of action. The results point to gossypol (S2303) as a promising antimicrobial compound that inhibits cell division by affecting FtsZ polymerization and has potential to be developed into an effective antimicrobial drug by chemical modification to minimize its cytotoxic effects in eukaryotic cells that were identified in this work.

Antibacterial activity of Chaga mushroom (Inonotus obliquus) extract against Staphylococcus aureus and Salmonella typhimurium: An In Vitro Study

Inonotus obliquus (Chaga mushroom) is a herbal and edible mushroom widely distributed in different regions, especially temperate regions. Chaga parasites on the trunk of angiosperms and grows, forming an irregular black mass. The medical importance of Chaga has been reported from ancient cultures to data due to its activity against a wide range of diseases. In the present study, the ethyl acetate extract of the Chaga mushroom has been tested against two bacterial pathogens, Staphylococcus aureus ATCC25923 and Salmonella typhimurium ATCC14028. The antimicrobial activity of the extract showed high activity with inhibition zone diameters of 25 mm for S.aureus and 28 mm for S.typhimurium and minimum inhibitory concentration (MICs) of 3.125 µg/ml for both bacterial pathogens. GC-MS analysis of the extract revealed several compounds with seven major compounds; they are 3-pentanone, propyl butyrate, Ethyl propionate, Trimethylsilymethanol, Neopentyl glycol, n-butyl acetate, and 1-butanol-3-methyl acetate. The antibacterial mode of action of Chaga extract was also elucidated by studying the effect of the extract on cell membrane permeability and recoded high effect on bacterial cell membrane permeability, which was confirmed by transmission electron microscope (TEM) images that revealed leakage of bacterial cell content outside the cells and death.