Recent Applications of Aggregation Induced Emission Probes for Antimicrobial Peptide Studies

Antimicrobial peptides (AMPs) are being intensively investigated as they are considered promising alternatives to antibiotics where their clinical efficacy is dwindling due to the emergence of antimicrobial resistance (AMR). Accompanying with the development of AMPs, a number of fluorescent probes have been developed to facilitate the understanding the modes of action of AMPs. These probes have been used to monitor the binding process, determine the working mechanism and evaluate the antimicrobial properties of AMPs. In particular, with the recent advance of aggregation-induced emission (AIE) fluorophores, that show many advantageous properties over traditional probes, there is an increasing research interest in using AIE probes for AMP studies. In this review, we give an overview of AMP development, highlight the recent progress of using fluorescence probes in particularly AIE probes in the AMP field and propose the future perspective of developing potent antimicrobial agents to combat AMR.     

The Effects of Natural Products and Environmental Conditions on Antimicrobial Resistance

Due to the extensive application of antibiotics in medical and farming practices, the continued diversification and development of antimicrobial resistance (AMR) has attracted serious public concern. With the emergence of AMR and the failure to treat bacterial infections, it has led to an increased interest in searching for novel antibacterial substances such as natural antimicrobial substances, including microbial volatile compounds (MVCs), plant-derived compounds, and antimicrobial peptides. However, increasing observations have revealed that AMR is associated not only with the use of antibacterial substances but also with tolerance to heavy metals existing in nature and being used in agriculture practice. Additionally, bacteria respond to environmental stresses, e.g., nutrients, oxidative stress, envelope stress, by employing various adaptive strategies that contribute to the development of AMR and the survival of bacteria. Therefore, we need to elucidate thoroughly the factors and conditions affecting AMR to take comprehensive measures to control the development of AMR.     

Selenium Nanomaterials to Combat Antimicrobial Resistance

The rise of antimicrobial resistance to antibiotics (AMR) as a healthcare crisis has led to a tremendous social and economic impact, whose damage poses a significant threat to future generations. Current treatments either are less effective or result in further acquired resistance. At the same time, several new antimicrobial discovery approaches are expensive, slow, and relatively poorly equipped for translation into the clinical world. Therefore, the use of nanomaterials is presented as a suitable solution. In particular, this review discusses selenium nanoparticles (SeNPs) as one of the most promising therapeutic agents based in the nanoscale to treat infections effectively. This work summarizes the latest advances in the synthesis of SeNPs and their progress as antimicrobial agents using traditional and biogenic approaches. While physiochemical methods produce consistent nanostructures, along with shortened processing procedures and potential for functionalization of designs, green or biogenic synthesis represents a quick, inexpensive, efficient, and eco-friendly approach with more promise for tunability and versatility. In the end, the clinical translation of SeNPs faces various obstacles, including uncertain in vivo safety profiles and mechanisms of action and unclear regulatory frameworks. Nonetheless, the promise possessed by these metalloid nanostructures, along with other nanoparticles in treating bacterial infections and slowing down the AMR crisis, are worth exploring.     

Utilizing Paper-Based Devices for Antimicrobial-Resistant Bacteria Detection

Antimicrobial resistance (AMR), the ability of a bacterial species to resist the action of an antimicrobial drug, has been on the rise due to the widespread use of antimicrobial agents. Per the World Health Organization, AMR has an estimated annual cost of USD 34 billion in the US and is predicted to be the number one cause of death worldwide by 2050. One way AMR bacteria can spread, and by which individuals can contract AMR infections, is through contaminated water. Monitoring AMR bacteria in the environment currently requires that samples be transported to a central laboratory for slow and labor intensive tests. We have developed an inexpensive assay using paper-based analytical devices (PADs) that can test for the presence of β-lactamase-mediated resistance. To demonstrate viability, the PAD was used to detect β-lactam resistance in wastewater and sewage and identified resistance in individual bacterial species isolated from environmental water sources.     

The upsurge of photocatalysts in antibiotic micropollutants treatment: Materials design,recovery, toxicity and bioanalysis

The excessive use of antimicrobial agents such as antibiotics and disinfectants for domestic purposes and industries polluted the water bodies severely in the recent past. Thus released antimicrobial agents negatively impact the environment and human health as it induce antimicrobial resistance (AMR) to microbes in the environment. Conventional biodegradation routes showed feasible antibiotics pollutants degradation. Nonetheless, they often demand a long time of operation (usually in days) and a major portion of the antimicrobial agents is left untreated unlike the complete oxidation with advanced oxidation processes. The residues of antibiotics left in the water bodies accelerate growth of microorganisms (bacterial, fungal, and viral) with AMR. In virtue of avoiding the catastrophe of widespread AMR, photocatalysis assisted antibiotic pollutant treatment is recently gaining a great popularity as an advanced oxidation process and has shown to be useful for the removal of antimicrobial compounds, mainly antibiotics. Recent review reports on photocatalytic antibiotic degradation focus on summarizing materials progress and antibiotics pollutants in chronological viewpoints. However, the relationship between photocatalytic materials and antibiotics oxidation reaction pathways and the toxicity of by-products are needed to be shown with better clarity to transfer the photocatalysis technique from lab to market in a safe way. This review critically analyzes the insights of energetic semiconductor structure lacking to achieve hydroxyl and superoxide radicals mediated antibiotics degradation, recommends new materials design (Z scheme) and standardization in the experimental designs, and also informs the influencing parameters on antibiotic degradation. It further assesses the possibility of recovering value-added chemicals from the photocatalytic treatment process and highlights the importance of environmental toxicity analysis. Overall, this review will be a resourceful guide for interdisciplinary researchers working on advanced photocatalysis and pharmaceutical pollutant treatment for achieving a sustainable ecology and initiating a circular economy in chemical industries.     

Utilizing Paper‐Based Devices for Antimicrobial‐Resistant Bacteria Detection

Antimicrobial resistance (AMR), the ability of a bacterial species to resist the action of an antimicrobial drug, has been on the rise due to the widespread use of antimicrobial agents. Per the World Health Organization, AMR has an estimated annual cost of USD 34 billion in the US and is predicted to be the number one cause of death worldwide by 2050. One way AMR bacteria can spread, and by which individuals can contract AMR infections, is through contaminated water. Monitoring AMR bacteria in the environment currently requires that samples be transported to a central laboratory for slow and labor intensive tests. We have developed an inexpensive assay using paper‐based analytical devices (PADs) that can test for the presence of β‐lactamase‐mediated resistance. To demonstrate viability, the PAD was used to detect β‐lactam resistance in wastewater and sewage and identified resistance in individual bacterial species isolated from environmental water sources.     

Interplay between amphiphilic peptides and nanoparticles for selective membrane destabilization and antimicrobial effects

As a result of an increasing number of bacteria developing resistance against antibiotics, antimicrobial peptides (AMPs) are attracting significant interest, particularly in relation to identification of peptides displaying potent but selective effects. Much less focus has been placed on delivery systems for AMPs, despite AMPs suffering from delivery challenges related to their size, cationicity, and amphiphilicity. Inorganic nanoparticles may provide opportunities for controlling peptide release, reducing infection-related AMP degradation, or increasing bioavailability. Numerous such nanomaterials display potent and triggerable antimicrobial effects on their own. When combined with AMPs, combinatorial and synergistic effects in relation to the behavior of such mixed systems as antimicrobials have been observed. The mechanistic origin of these effects are poorly understood that at present, however, precluding rational design of mixed nanoparticle antimicrobials/AMPs and nanoparticulate delivery systems for AMPs. Here, the area of membrane interactions and antimicrobial effects of inorganic nanomaterials are briefly outlined, in combination with AMPs.     

Nano-enabled Quenching of Bacterial Communications for the Prevention of Biofilm Formation

Biofilm formation is a major threat to industry, the environment and human health. While killing of embedded microbes in biofilms may inevitably lead to the evolution of antimicrobial resistance (AMR), catalytic quenching of bacterial communications by lactonase is a promising antifouling approach. Given the shortcomings of protein enzymes, it is attractive to engineer synthetic materials to mimic the activity of lactonase. Herein, an efficient lactonase-like Zn−N_x−C nanomaterial was synthesized by tuning the coordination environment around zinc atoms to mimic the active domain of lactonase for catalytical interception of bacterial communications in biofilm formation. The Zn−N_x−C material could selectively catalyze 77.5 % hydrolysis of N-acylated-L-homoserine lactone (AHL), a critical bacterial quorum sensing (QS) signal in biofilm construction. Consequently, AHL degradation downregulated the expression of QS-related genes in antibiotic resistant bacteria and significantly prevented biofilm formation. As a proof of concept, Zn−N_x−C-coated iron plates prevented 80.3 % biofouling after a month exposure in river. Overall, our study provides a nano-enabled contactless antifouling insight to avoid AMR evolution by engineering nanomaterials for mimicking the key bacterial enzymes (e.g., lactonase) functioning in biofilm construction.     

Carbon nanomaterials in microbial sensing and bactericidal applications

The spread of antimicrobial resistance and lesser development of new antibiotics have intensified the search for new antimicrobial and diagnostic vehicles. Carbon nanomaterials (CNMs), which broadly include carbon dots, carbon nanotubes, and graphene/graphene oxide nanostructures, have emerged as promising theranostic materials exhibiting, in many instances, potent antibacterial activities and diagnostic capabilities. Ease of synthesis, tunable physicochemical properties, biocompatibility, and diverse modes of action make CNMs a powerful class of theranostic nanomaterials. This review discusses recent studies illuminating innovative new CNMs and their applications in bacterial theranostics. We particularly emphasize the relationship between the structural parameters and overall chemical properties of CNMs and their biological impact and utilization. Overall, the expanding work on the development and use of CNMs in therapeutic, sensing, and diagnostic applications in the microbial world underscores the considerable potential of these nanomaterials.     

Mass spectrometry profiling of single bacterial cells reveals metabolic regulation during antibiotics induced bacterial filamentation

Bacterial antimicrobial resistance (AMR) is a severe threat to global health and development. Under the stimulation of antibiotics, bacterial cells can undergo filamentation and generate daughter cells with stronger AMR. The current research on bacterial AMR mechanism is mainly conducted with a population of cells. However, bacterial cells exhibit heteroresistance, making the study at population level not reliable. Herein, we developed single bacterial cell metabolic profiling by mass spectrometry (MS) to study bacterial AMR at single-cell level. By utilizing a microprobe controlled by a microoperation platform, single filamentous extended spectrum beta-lactamase (ESBL) producing Escherichia coli (ESBL-E. coli) cells generated by ceftriaxone sodium stimulation can be extracted and spray-ionized for MS analysis. Heterogeneous among ESBL-E. coli cells under the same antibiotic stimulus condition was observed from mass spectra as well as cell morphology. The metabolic profiles by MS of different individual cells can be clustered into subgroups well in accordance with bacterial cell length. Metabolic pathways including arginine and proline metabolism, as well as cysteine and methionine metabolism were disclosed to play an important role in the bacterial SOS-associated filamentation against antibiotics. The microprobe electrospray ionization-MS-based single bacterial cell analysis method is promising in the study of various bacterial AMR mechanism and can reveal the heterogeneity of bacterial AMR from-cell-to-cell.     

Emerging markers for antimicrobial resistance monitoring

The appearance and spread of antibiotic-resistant pathogens known as antimicrobial resistance (AMR) is one of the major worldwide health crises that humanity have to deal with over the next decades. One of the main methods for addressing AMR is the effective screening for antimicrobial insensitivity in clinical and environmental monitoring. Current clinical laboratory procedures use traditional culture-based antibiotic susceptibility testing (AST) methods, which can take up to 24 h to identify which drug is suitable for the infection inhibition. Therefore, it is vital to develop novel strategies that offer quick, simple, affordable, reliable, sensitive and accurate AMR monitoring. Sensors for AMR markers detection could possess the essential qualities for quickly identifying resistant microorganisms and could give vital data for the selection of antibacterial drugs administration. This review offers a summary of the innovative application of these AMR markers detection strategies focusing on healthcare and environmental surveillance for the AMR genotypic or phenotypic assessment.     

Cheminformatic Characterization of Natural Antimicrobial Products for the Development of New Lead Compounds

The growing antimicrobial resistance (AMR) of pathogenic organisms to currently prescribed drugs has resulted in the failure to treat various infections caused by these superbugs. Therefore, to keep pace with the increasing drug resistance, there is a pressing need for novel antimicrobial agents, especially from non-conventional sources. Several natural products (NPs) have been shown to display promising in vitro activities against multidrug-resistant pathogens. Still, only a few of these compounds have been studied as prospective drug candidates. This may be due to the expensive and time-consuming process of conducting important studies on these compounds. The present review focuses on applying cheminformatics strategies to characterize, prioritize, and optimize NPs to develop new lead compounds against antimicrobial resistance pathogens. Moreover, case studies where these strategies have been used to identify potential drug candidates, including a few selected open-access tools commonly used for these studies, are briefly outlined.     

Assays Evaluating Antimicrobial Activity of Nanoparticles: A Myth Buster

The kingdom of interdisciplinary research is advancing forcefully; nanotechnology is one discipline that has impressed biologist, physicist, chemists, engineers and physicians. For studying the antimicrobial activity of nanomaterials, standard protocols that were used earlier for assessing the efficiency of antibiotics and compounds were taken for granted. The disc diffusion and well diffusion methods were ideally designed for studying bactericidal activity of diffusible compounds, mainly antibiotics and not for a particulate interaction based systems. A mere extrapolation of the testing method for testing nanoparticles, with an exclusion of the plate count method, appears to have stepped out of the fundamental principle whereby nanomaterials and microbes are required to interact. This report highlights the flaw in the results obtained from using disc/well diffusion methods for determining the bioactivity of nanomaterials, owing to the physical barrier between bacteria and the nanoparticles. A more realistic and reliable method, where the nanoparticles and microbial cells are put into an open base where they can freely interact is mandatory to assess toxicity/compatibility of nanomaterials. Failure in utilizing the apt testing mode, may eventually lead to declaring toxic nanomaterials as non-toxic and underestimating the antimicrobial ability of few other potent nanomaterials.     

Design principles for bacteria-responsive antimicrobial nanomaterials

Despite advancements in biomedical sciences and medicine, bacterial infection remains a leading cause of death globally. The conventional treatment of bacterial infections through general administration of antibiotics has been challenged by the emergence of antibiotics resistance. An exciting direction to solve current challenges that attracted enormous interest in the last decade is focused on designing stimuli-responsive systems incorporating a wide range of antimicrobial nanomaterials. The aim of this review is to highlight fundamental principles involved in the design of bacteria-responsive nanosystems that release their antibacterial load in response only to the specific environment and factors produced endogenously by bacteria. Such specific changes to the micro-environment include changes in pH, reactive oxygen species, and production of enzymes specific to bacteria. We provide examples and a critical review of such systems and finish with the authors’ perspective for the future of the field.     

Approaches for Mitigating Microbial Biofilm-Related Drug Resistance: A Focus on Micro- and Nanotechnologies

Biofilms play an essential role in chronic and healthcare-associated infections and are more resistant to antimicrobials compared to their planktonic counterparts due to their (1) physiological state, (2) cell density, (3) quorum sensing abilities, (4) presence of extracellular matrix, (5) upregulation of drug efflux pumps, (6) point mutation and overexpression of resistance genes, and (7) presence of persister cells. The genes involved and their implications in antimicrobial resistance are well defined for bacterial biofilms but are understudied in fungal biofilms. Potential therapeutics for biofilm mitigation that have been reported include (1) antimicrobial photodynamic therapy, (2) antimicrobial lock therapy, (3) antimicrobial peptides, (4) electrical methods, and (5) antimicrobial coatings. These approaches exhibit promising characteristics for addressing the impending crisis of antimicrobial resistance (AMR). Recently, advances in the micro- and nanotechnology field have propelled the development of novel biomaterials and approaches to combat biofilms either independently, in combination or as antimicrobial delivery systems. In this review, we will summarize the general principles of clinically important microbial biofilm formation with a focus on fungal biofilms. We will delve into the details of some novel micro- and nanotechnology approaches that have been developed to combat biofilms and the possibility of utilizing them in a clinical setting.     

Identification and design of antimicrobial peptides for therapeutic applications

Indiscriminate use of antibiotics has led to a rapid increase of antibiotic resistance among microbes which has increased the need to develop novel antimicrobial agents to fight various infectious diseases. Peptide antibiotics signify a novel class of therapeutic agents and have been isolated from a wide variety of multi-cellular organisms. Peptide antibiotics have shown broad-spectrum antimicrobial activity and they not only kill different bacteria, but also kill various fungi, parasites, protozoans and cancerous cells. Peptides bear several properties that make them particularly attractive such as their small size, rapid activity and a low chance for development of resistance. Because of these distinct properties, the focus for research on antimicrobial peptides has increased tremendously in the recent years. Despite their potential, only selected cationic antimicrobial peptides have been able to enter in clinical trials. Therefore, there is a pressing need to develop new approaches to identify novel antimicrobial peptide therapeutics replacing conventional antibiotics. Recent findings strongly suggest that one can design a new generation of antimicrobials peptides with a wide range of systemic and topical applications against bacterial infections. In this review, we focus on the identification and design of novel antimicrobial peptides for therapeutic applications based on different approaches and strategies. This review also highlights some recent advances in the study of the molecular basis of anti-microbial activity in these peptides, their current pharmacological and clinical development and future directions and applications.     

Broad spectrum antibacterial zinc oxide-reduced graphene oxide nanocomposite for water depollution

Antimicrobial-resistance (AMR) is a global health challenge arising from the evolution of bacteria, viruses, fungi, and parasites, such that pathogenic microorganisms no longer respond to classical therapies. AMR and the rise of so-called ‘superbugs’ requires innovative nanomaterials and biostatic strategies. Here we report a broad spectrum, antimicrobial nanomaterial integrating light-responsive ZnO nanoparticles (NP) and reduced graphene oxide (rGO) into a heterojunction semiconductor nanocomposite for water depollution. Simultaneous chemical reduction of Zn sulphate and GO sheets yields a low concentration (0.5 mol%) of 10 nm ZnO nanoparticles decorating fragmented rGO nanosheets, with a total surface area of 12 m²/g and optical band gap of 1.6 eV. Antimicrobial performance of the ZnO-rGO nanocomposite was evaluated against methicillin-resistant Staphylococcus aureus (MRSA), Escherichia coli 0157:H7 and Salmonella typhimurium bacteria, which are prevalent in contaminated aquatic systems; antimicrobial efficacy against these organisms was 96%, 97%, and 73%, respectively, for a loading of 2 mg/mL, evidencing a strong synergy compared with pure ZnO or rGO components. ZnO-rGO was also an effective photocatalyst for the aqueous degradation of Malachite Green dye, suggesting that its mode of antibacterial action reflects the production of reactive oxygen species under ambient illumination.     

Semi-Rational Engineering of Leucine Dehydrogenase for <Emphasis Type="Italic">L</Emphasis>-2-Aminobutyric Acid Production

Rapid increase in antibiotic resistance has posed a worldwide threat, due to increased mortality, morbidity, and expenditure caused by antibiotic-resistant microbes. Recent development of the antimicrobial peptides like viscotoxin (Vt) has been successfully comprehended as a substitute for classical antibiotics. A structurally stable peptide, Vt can enhance antimicrobial property and can be used for various developmental purposes. Thus, structural stability among the antimicrobial peptides, Vt A1 (3C8P), A2 (1JMN), A3 (1ED0), B (1JMP), and C (1ORL) of Viscus album was computationally analyzed. In specific, the static confirmation of VtA3 showed high number of intramolecular interactions, along with an increase in hydrophobicity than others comparatively. Further, conformational sampling was used to analyze various geometrical parameters such as root mean square deviation, root mean square fluctuation, radius of gyration, and ovality which also revealed the structural stability of VtA3. Moreover, the statistically validated contours of surface area, lipophilicity, and distance constraints of disulfide bonds also supported the priority of VtA3 with respect to stability. Finally, the functional activity of peptides was accessed by computing their free energy of membrane association and membrane interactions, which defined VtA3 as functionally stable. Currently, peptide-based antibiotics and nanoparticles have attracted the pharmaceutical industries for their potential therapeutic applications. Thereby, it is proposed that viscotoxin A3 (1ED0) could be used as a preeminent template for scaffolding potentially efficient antimicrobial peptide-based drugs and nanomaterials in future.     

Design of Photosensitizing Agents for Targeted Antimicrobial Photodynamic Therapy

Photodynamic inactivation of microorganisms has gained substantial attention due to its unique mode of action, in which pathogens are unable to generate resistance, and due to the fact that it can be applied in a minimally invasive manner. In photodynamic therapy (PDT), a non-toxic photosensitizer (PS) is activated by a specific wavelength of light and generates highly cytotoxic reactive oxygen species (ROS) such as superoxide (O²⁻, type-I mechanism) or singlet oxygen (¹O₂*, type-II mechanism). Although it offers many advantages over conventional treatment methods, ROS-mediated microbial killing is often faced with the issues of accessibility, poor selectivity and off-target damage. Thus, several strategies have been employed to develop target-specific antimicrobial PDT (aPDT). This includes conjugation of known PS building-blocks to either non-specific cationic moieties or target-specific antibiotics and antimicrobial peptides, or combining them with targeting nanomaterials. In this review, we summarise these general strategies and related challenges, and highlight recent developments in targeted aPDT.     

Facet-dependent antibacterial activity of Au nanocrystals

Engineered nanomaterials have attracted significantly attention as one of the most promising antimicrobial agents for against multidrug resistant infections. The toxicological responses of nanomaterials are closely related to their physicochemical properties, and establishment of a structure-activity relationship for nanomaterials at the nano-bio interface is of great significance for deep understanding antibacterial toxicity mechanisms of nanomaterials and designing safer antibacterial nanomaterials. In this study, the antibacterial behaviors of well-defined crystallographic facets of a series of Au nanocrystals, including {100}-facet cubes, {110}-facet rhombic dodecahedra, {111}-facet octahedra, {221}-facet trisoctahedra and {720}-facet concave cubes, was investigated, using the model bacteria Staphylococcus aureus. We find that Au nanocrystals display substantial facet-dependent antibacterial activities. The low-index facets of cubes, octahedra, and rhombic dodecahedra show considerable antibacterial activity, whereas the high-index facets of trisoctahedra and concave cubes remained inert under biological conditions. This result is in stark contrast to the previous paradigm that the high-index facets were considered to have higher bioactivity as compared with low-index facets. The antibacterial mechanism studies have shown that the facet-dependent antibacterial behaviors of Au nanocrystals are mainly caused by differential bacterial membrane damage as well as inhibition of cellular enzymatic activity and energy metabolism. The faceted Au nanocrystals are unique in that they do not induce generation of reactive oxygen species, as validated for most antibiotics and antimicrobial nanostructures. Our findings may provide a deeper understanding of facet-dependent toxicological responses and suggest the complexities of the nanomaterial-cell interactions, shedding some light on the development of high performance Au nanomaterials-based antibacterial therapeutics.