Contribution of Invariant Natural Killer T Cells to the Clearance of Pseudomonas aeruginosa from Skin Wounds

Chronic infections are considered one of the most severe problems in skin wounds, and bacteria are present in over 90% of chronic wounds. Pseudomonas aeruginosa is frequently isolated from chronic wounds and is thought to be a cause of delayed wound healing. Invariant natural killer T (iNKT) cells, unique lymphocytes with a potent regulatory ability in various inflammatory responses, accelerate the wound healing process. In the present study, we investigated the contribution of iNKT cells in the host defense against P. aeruginosa inoculation at the wound sites. We analyzed the re-epithelialization, bacterial load, accumulation of leukocytes, and production of cytokines and antimicrobial peptides. In iNKT cell–deficient (Jα18KO) mice, re-epithelialization was significantly decreased, and the number of live colonies was significantly increased, when compared with those in wild-type (WT) mice on day 7. IL-17A, and IL-22 production was significantly lower in Jα18KO mice than in WT mice on day 5. Furthermore, the administration of α-galactosylceramide (α-GalCer), a specific activator of iNKT cells, led to enhanced host protection, as shown by reduced bacterial load, and to increased production of IL-22, IL-23, and S100A9 compared that of with WT mice. These results suggest that iNKT cells promote P. aeruginosa clearance during skin wound healing.     

Acquisition and Spread of Antimicrobial Resistance: A tet(X) Case Study

Understanding the mechanisms leading to the rise and dissemination of antimicrobial resistance (AMR) is crucially important for the preservation of power of antimicrobials and controlling infectious diseases. Measures to monitor and detect AMR, however, have been significantly delayed and introduced much later after the beginning of industrial production and consumption of antimicrobials. However, monitoring and detection of AMR is largely focused on bacterial pathogens, thus missing multiple key events which take place before the emergence and spread of AMR among the pathogens. In this regard, careful analysis of AMR development towards recently introduced antimicrobials may serve as a valuable example for the better understanding of mechanisms driving AMR evolution. Here, the example of evolution of tet(X), which confers resistance to the next-generation tetracyclines, is summarised and discussed. Initial mechanisms of resistance to these antimicrobials among pathogens were mostly via chromosomal mutations leading to the overexpression of efflux pumps. High-level resistance was achieved only after the acquisition of flavin-dependent monooxygenase-encoding genes from the environmental microbiota. These genes confer resistance to all tetracyclines, including the next-generation tetracyclines, and thus were termed tet(X). ISCR2 and IS26, as well as a variety of conjugative and mobilizable plasmids of different incompatibility groups, played an essential role in the acquisition of tet(X) genes from natural reservoirs and in further dissemination among bacterial commensals and pathogens. This process, which took place within the last decade, demonstrates how rapidly AMR evolution may progress, taking away some drugs of last resort from our arsenal.     

Microbial susceptibility of various polymers and evaluation of thermoplastic elastomers with antimicrobial additives

The incursion of microbial growth on polymeric products can deteriorate their performance and lead to the development of undesirable staining and odors. A growing trend in the industry has aimed to reduce microbial populations on high-touch surfaces via the use of antimicrobials to protect material aesthetics and durability or to prevent the spread of pathogenic microorganisms. In this study, a variety of plastic substrates (30 unique polymer compounds), including poly(acrylonitrile-co-butadiene-co-styrene), poly(butylene terephthalate), poly(etherimide), various thermoplastic elastomers (TPEs), poly(carbonates), and poly(amides), were screened for susceptibility to microbial attack using American Society for Testing and Materials (ASTM) G21 (fungi susceptibility), Japanese Industrial Standard (JIS) Z2801, and modified ASTM E1428-15a (bacterial susceptibility) test standards. TPEs were determined to be most susceptible to microbial attack under the appropriate environmental conditions. Subsequent studies assessed the use of an antimicrobial additive, zinc pyrithione (ZPT), for potential efficacy in a variety of TPE blends for diverse target market applications. ZPT proved to be very effective in protecting TPEs, reducing Staphylococcus aureus and Escherichia coli populations by 99.9% or more in JIS Z2801 testing and inhibiting fungal growth (rating = 0) according to the ASTM G21 standard.     

Bioinspired Supramolecular Slippery Organogels for Controlling Pathogen Spread by Respiratory Droplets

Surface-deposited pathogens are sources for the spread of infectious diseases. Protecting public facilities with a replaceable or recyclable antifouling coating is a promising approach to control pathogen transmission. However, most antifouling coatings are less effective in preventing pathogen-contained respiratory droplets because these tiny droplets are difficult to repel, and the deposited pathogens can remain viable from hours to days. Inspired by mucus, an antimicrobial supramolecular organogel for the control of microdroplet-mediated pathogen spread is developed. The developed organogel coating harvests a couple of unique features including localized molecular control-release, readily damage healing, and persistent fouling-release properties, which are preferential for antifouling coating. Microdroplets deposited on the organogel surfaces will be spontaneously wrapped with a thin liquid layer, and will therefore be disinfected rapidly due to a mechanism of spatially enhanced release of bactericidal molecules. Furthermore, the persistent fouling-release and damage-healing properties will significantly extend the life-span of the coating, making it promising for diverse applications.     

Aminoacyl-tRNA Synthetases as Valuable Targets for Antimicrobial Drug Discovery

Aminoacyl-tRNA synthetases (aaRSs) catalyze the esterification of tRNA with a cognate amino acid and are essential enzymes in all three kingdoms of life. Due to their important role in the translation of the genetic code, aaRSs have been recognized as suitable targets for the development of small molecule anti-infectives. In this review, following a concise discussion of aaRS catalytic and proof-reading activities, the various inhibitory mechanisms of reported natural and synthetic aaRS inhibitors are discussed. Using the expanding repository of ligand-bound X-ray crystal structures, we classified these compounds based on their binding sites, focusing on their ability to compete with the association of one, or more of the canonical aaRS substrates. In parallel, we examined the determinants of species-selectivity and discuss potential resistance mechanisms of some of the inhibitor classes. Combined, this structural perspective highlights the opportunities for further exploration of the aaRS enzyme family as antimicrobial targets.     

Antithrombin and Its Role in Host Defense and Inflammation

Antithrombin (AT) is a natural anticoagulant that interacts with activated proteases of the coagulation system and with heparan sulfate proteoglycans (HSPG) on the surface of cells. The protein, which is synthesized in the liver, is also essential to confer the effects of therapeutic heparin. However, AT levels drop in systemic inflammatory diseases. The reason for this decline is consumption by the coagulation system but also by immunological processes. Aside from the primarily known anticoagulant effects, AT elicits distinct anti-inflammatory signaling responses. It binds to structures of the glycocalyx (syndecan-4) and further modulates the inflammatory response of endothelial cells and leukocytes by interacting with surface receptors. Additionally, AT exerts direct antimicrobial effects: depending on AT glycosylation it can bind to and perforate bacterial cell walls. Peptide fragments derived from proteolytic degradation of AT exert antibacterial properties. Despite these promising characteristics, therapeutic supplementation in inflammatory conditions has not proven to be effective in randomized control trials. Nevertheless, new insights provided by subgroup analyses and retrospective trials suggest that a recommendation be made to identify the patient population that would benefit most from AT substitution. Recent experiment findings place the role of various AT isoforms in the spotlight. This review provides an overview of new insights into a supposedly well-known molecule.     

Practical in-storage interventions to control foodborne pathogens on fresh produce

Although tremendous efforts have been made to ensure fresh produce safety, various foodborne outbreaks and recalls occur annually. Most of the current intervention strategies are evaluated within a short timeframe (less than 1 h), leaving the behavior of the remaining pathogens unknown during subsequent storages. This review summarized outbreak and recall surveillance data from 2009 to 2018 obtained from government agencies in the United States to identify major safety concerns associated with fresh produce, discussed the postharvest handling of fresh produce and the limitations of current antimicrobial interventions, and reviewed the intervention strategies that have the potential to be applied in each storage stage at the commercial scale. One long-term (up to 12 months) prepacking storage (apples, pears, citrus among others) and three short-term (up to 3 months) postpacking storages were identified. During the prepacking storage, continuous application of gaseous ozone at low doses (≤1 ppm) is a feasible option. Proper concentration, adequate circulation, as well as excess gas destruction and ventilation systems are essential to commercial application. At the postpacking storage stages, continuous inhibition can be achieved through controlled release of gaseous chlorine dioxide in packaging, antimicrobial edible coatings, and biocontrol agents. During commercialization, factors that need to be taken into consideration include physicochemical properties of antimicrobials, impacts on fresh produce quality and sensory attributes, recontamination and cross-contamination, cost, and feasibility of large-scale production. To improve fresh produce safety and quality during storage, the collaboration between researchers and the fresh produce industry needs to be improved.     

Modification of narrow‐spectrum peptidomimetic polyurethanes with fatty acid chains confers broad‐spectrum antibacterial activity

Each year, thousands of patients die from antimicrobial‐resistant bacterial infections that fail to respond to conventional antibiotic treatment. Antimicrobial polymers are a promising new method of combating antibiotic‐resistant bacterial infections. We have previously reported the synthesis of a series of narrow‐spectrum peptidomimetic antimicrobial polyurethanes that are effective against Gram‐negative bacteria, such as Escherichia coli; however, these polymers are not effective against Gram‐positive bacteria, such as Staphylococcus aureus. With the aim of understanding the correlation between chemical structure and antibacterial activity, we have subsequently developed three structural variants of these antimicrobial polyurethanes using post‐polymerization modification with decanoic acid and oleic acid. Our results show that such modifications converted the narrow‐spectrum antibacterial activity of these polymers into broad‐spectrum activity against Gram‐positive species such as S. aureus, however, also increasing their toxicity to mammalian cells. Mechanistic studies of bacterial membrane disruption illustrate the differences in antibacterial action between the various polymers. The results demonstrate the challenge of balancing antimicrobial activity and mammalian cell compatibility in the design of antimicrobial polymer compositions. © 2019 Society of Chemical Industry     

Peptide-Mimicking Poly(2-oxazoline)s Displaying Potent Antimicrobial Properties

Poly(2-oxazoline)s have excellent biocompatibility and have been used as FDA-approved indirect food additives. The inert property of the hydrophilic poly(2-oxazoline)s suggests them as promising substitutes for poly(ethylene glycol) (PEG) in various applications such as anti-biofouling agents. It was recently reported that poly(2-oxazoline)s themselves have antimicrobial properties as synthetic mimics of host defense peptides. These studies revealed the bioactive properties of poly(2-oxazoline)s as a new class of functional peptide mimics, by mimicking host defense peptides to display potent and selective antimicrobial activities against methicillin-resistant Staphylococcus aureus both in vitro and in vivo, without concerns about antimicrobial resistance. The high structural diversity, facile synthesis, and potent and tunable antimicrobial properties underscore the great potential of poly(2-oxazoline)s as a class of novel antimicrobial agents in dealing with drug-resistant microbial infections and antimicrobial resistance.     

Antimicrobial and Anticancer Properties of Synthetic Peptides Derived from the Wasp Parachartergus fraternus

Agelaia-MPI and protonectin are antimicrobial peptides isolated from the wasp Parachartergus fraternus that show antimicrobial and neuroactive activities. Previously, two analogues of these peptides, neuroVAL and protonectin-F, were designed to reduce nonspecific toxicity and improve potency. Here, the three-dimensional structures of neuroVAL, protonectin and protonectin-F were determined by using circular dichroism and NMR spectroscopy. Antibacterial, antifungal, cytotoxic and hemolytic activities were tested for the parent peptides and analogues. All peptides showed moderate antimicrobial activity against Gram-positive bacteria, with agelaia-MPI being the most active. Protonectin and protonectin-F were found to be toxic to cancerous and noncancerous cell lines. Internalization experiments revealed that these peptides accumulate inside both cell types. By contrast, neuroVAL was nontoxic to all tested cells and was able to enter cells without accumulating. In summary, neuroVAL has potential as a nontoxic cell-penetrating peptide, while protonectin-F needs further modification to realize its potential as an antitumor peptide.