Beyond Traditional Antimicrobials: A Caenorhabditis elegans Model for Discovery of Novel Anti-infectives

The spread of antibiotic resistance amongst bacterial pathogens has led to an urgent need for new antimicrobial compounds with novel modes of action that minimize the potential for drug resistance. To date, the development of new antimicrobial drugs is still lagging far behind the rising demand, partly owing to the absence of an effective screening platform. Over the last decade, the nematode Caenorhabditis elegans has been incorporated as a whole animal screening platform for antimicrobials. This development is taking advantage of the vast knowledge on worm physiology and how it interacts with bacterial and fungal pathogens. In addition to allowing for in vivo selection of compounds with promising anti-microbial properties, the whole animal C. elegans screening system has also permitted the discovery of novel compounds targeting infection processes that only manifest during the course of pathogen infection of the host. Another advantage of using C. elegans in the search for new antimicrobials is that the worm itself is a source of potential antimicrobial effectors which constitute part of its immune defense response to thwart infections. This has led to the evaluation of effector molecules, particularly antimicrobial peptides (AMPs), as candidates for further development as therapeutic agents. In this review, we provide an overview on the use of the C. elegans model for identification of novel anti-infectives. We highlight some highly potential lead compounds obtained from C. elegans-based screens, particularly those that target bacterial virulence or host defense to eradicate infections, a mechanism distinct from the action of conventional antibiotics. We also review the prospect of using C. elegans AMPs as an antimicrobial strategy to treat infections

Extrusion-Based Bioprinted Boron Nitride Nanotubes Reinforced Alginate Scaffolds: Mechanical, Printability and Cell Viability Evaluation

Alginate (Alg) hydrogels are commonly used as bioinks in 3D bioprinting. However, one of the significant drawbacks of using Alg hydrogels is their unstable mechanical properties. In this study, a novel hydrogel-based ink composed of Alg reinforced with functionalised boron nitride nanotubes (f-BNNTs) was developed and systematic quantitative characterisation was conducted to validate its printability, physiochemical properties and biocompatibility. The printability, contact angle and mechanical test results indicated good structural stability of the scaffolds. The thermal stability of the scaffolds increased with the incorporation of f-BNNTs into Alg. Human embryonic kidney cells (HEK 293T) were seeded on the scaffolds and the cell viability was recorded for 24, 48 and 72 h. Quantitative studies showed a slight effect on toxicity with a higher concentration of BNNTs in scaffolds. The results suggest that the 3D printable f-BNNTs reinforced Alg could be used as bioink for tissue engineering applications with further studies on biocompatibility

Orthosiphon stamineus protects Caenorhabditis elegans against Staphylococcus aureus infection through immunomodulation

Amidst growing concerns over the spread of antibiotic-resistant Staphylococcus aureus strains, the identification of alternative therapeutic molecules has become paramount. Previously, we utilized a Caenorhabditis elegans–S. aureus screening platform to identify potential anti-infective agents from a collection of natural extracts and synthetic compounds. One of the hits obtained from the screen was the aqueous extract of Orthosiphon stamineus leaves (UE-12) that enhanced the survival of infected nematodes without interfering with bacterial growth. In this study, we used a fluorescent transgenic reporter strain and observed that the repressed expression of the lys-7 defense gene in infected nematodes was restored in the presence of UE-12. Analysis of a selected panel of PMK-1 and DAF-16-regulated transcripts and loss-of-function mutants in these pathways indicates that the protective role of UE-12 is mediated via the p38 MAP kinase and insulin-like signaling pathways. Further analysis of a panel of known bioactive compounds of UE-12 proposed eupatorin (C18H16O7) as the possible candidate active molecule contributing to the anti-infective property of UE-12. Taken together, these findings strongly suggest that the O. stamineus leaf extract is a promising anti-infective agent that confers an advantage in survival against S. aureus infection by modulating the immune response of the infected host

Targeting Staphylococcus aureus Toxins: A Potential form of Anti-Virulence Therapy

Staphylococcus aureus is an opportunistic pathogen and the leading cause of a wide range of severe clinical infections. The range of diseases reflects the diversity of virulence factors produced by this pathogen. To establish an infection in the host, S. aureus expresses an inclusive set of virulence factors such as toxins, enzymes, adhesins, and other surface proteins that allow the pathogen to survive under extreme conditions and are essential for the bacteria’s ability to spread through tissues. Expression and secretion of this array of toxins and enzymes are tightly controlled by a number of regulatory systems. S. aureus is also notorious for its ability to resist the arsenal of currently available antibiotics and dissemination of various multidrug-resistant S. aureus clones limits therapeutic options for a S. aureus infection. Recently, the development of anti-virulence therapeutics that neutralize S. aureus toxins or block the pathways that regulate toxin production has shown potential in thwarting the bacteria’s acquisition of antibiotic resistance. In this review, we provide insights into the regulation of S. aureus toxin production and potential anti-virulence strategies that target S. aureus toxins

Extrusion of Cell Encapsulated in Boron Nitride Nanotubes Reinforced Gelatin—Alginate Bioink for 3D Bioprinting

Three-dimensional (3D) bioprinting, an innovative technology, has gained the attention of researchers as a promising technique for the redevelopment of complex tissue or organ structures. Despite significant advancements, a major challenge in 3D bioprinting is the limited number of suitable bioinks that fulfil the physiochemical requirements to produce complicated structures. Therefore, there is a demand for the production of bioinks for 3D bioprinting techniques. In this short communication, THP-1 cells encapsulated in boron nitride nanotubes (BNNTs) reinforced gelatin and alginate bioink was prepared. The study investigated the impact on the cells during printing using a fluorescence cell image. The results showed that the pure polymer bioinks demonstrated poor printability properties with the incorporation of cells. However, BNNT-combined bioink showed a significant increase in structural integrity even after the incorporation of cells. Furthermore, the scaffold structure was successfully printed with the cells incorporated bioink, and a considerable number of live cells were observed. With further studies, BNNTs as a promising nanomaterial for formulating bioink encapsulated with cells can be understood fully

Detection of Burkholderia pseudomallei toxin-mediated inhibition of protein synthesis using a Caenorhabditis elegans ugt-29 biosensor

International audienceToxins are believed to play a crucial role in Burkholderia pseudomallei pathogenicity, however to date, only a few have been identified. The discovery of additional toxic molecules is limited by the lack of a sensitive indicator of B. pseudomallei toxicity. Previously, from a whole genome transcriptome analysis of B. pseudomallei-infected Caenorhabditis elegans, we noted significant overexpression of a number of worm genes encoding detoxification enzymes, indicating the host's attempt to clear bacterial toxic molecules. One of these genes, ugt-29, a family member of UDP-glucuronosyltransferases, was the most robustly induced phase II detoxification gene. In this study, we show that strong induction of ugt-29 is restricted to infections by the most virulent species among the pathogens tested. We also noted that ugt-29 is activated upon disruption of host protein synthesis. Hence, we propose that UGT-29 could be a promising biosensor to detect B. pseudomallei toxins that compromise host protein synthesis. The identification of bactobolin, a polyketide-peptide hybrid molecule, as a toxic molecule of B. pseudomallei further verifies the utilization of this surveillance system to search for bacterial toxins. Hence, a ugt-29 based reporter should be useful in screening for other molecules that inhibit host protein synthesis