Microbial flow cytometry: An ideal tool for prospective antimicrobial drug development

doi:10.1016/j.ab.2016.05.025

Analytical Biochemistry

Volume 509, 15 September 2016, Pages 89-91

https://doi.org/10.1016/j.ab.2016.05.025 Get rights and content

Abstract

Flow cytometry has tremendous applications in qualitative and quantitative analysis of characteristics of single microbial cells. Its ability to efficiently discriminate and quantify multiple parameters of microbial cells has made it a powerful tool to catalog the mechanism of action of antimicrobial peptides (AMPs) on target cells. Here, we provide a comprehensive overview and strategic design on how multi-parametric analysis of flow cytometry is unsurpassed in studying the antimicrobial process of AMPs in an accurate and rapid way. This strategy provides a conceptual framework for understanding distinct classes of AMPs and getting insights into antimicrobial mechanisms of novel AMPs.

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Cited by (4)

Current strategies to determine antifungal and antimicrobial activity of natural compounds
2021, Microbiological Research
Citation Excerpt :
This ability makes it possible to explore unique functional mechanisms of AMPs, natural occurring molecules that are important components of a host defense system and have great potential to serve as a basis for a new antimicrobial drug class targeting infectious pathogens that are difficult to treat, with unsurpassed sensitivity and accuracy compared with other techniques. A study where optimized CFM was used to examine the antimicrobial process of AMPs resulted in a highly suitability of the assay for the rapid and precise understanding of the mechanism and could have a great impact on antimicrobial drug development (Balouiri et al., 2016; Muthirulan and Chandrasekaran, 2016). Brown et al. (2019) used the FCM to quantify bacteria and estimate the biomass in activated sludge.
Fungal and microbial infections are increasingly common diseases affecting not only humans, but also animals. Despite the fact that there are wide ranges of antifungal drugs that can be used as therapy against different types of mycosis, the large-scale needed for new antifungal and antimicrobial agents is undeniable. The reasons for a great demand for new agents are low effectiveness due to the development of resistance, host toxicity and various side effects of currently used therapeutics. In order to develop novel drugs against fungal infections, scientists need to search for new molecules that show antimicrobial activity. However, there are various methods to determine antifungal and antimicrobial activity such as diffusion methods, bioautography methods, dilution methods and other frequently used methods. This review aims to explain the methodologies mentioned, to highlight the functioning, usage, advantages and disadvantages and to compare the techniques using different sources of the last years. Additionally, some of the currently investigated natural compounds such as essential oils, which show promising results in the medication of fungal diseases, are mentioned.
The structure-mechanism relationship and mode of actions of antimicrobial peptides: A review
2021, Trends in Food Science and Technology
Citation Excerpt :
According to different strategies, flow cytometry can figure out the membrane acting AMP and cell-translocating AMP, membrane acting AMP can be sub-divided into membrane pore-forming AMPS and membrane disrupting AMPs. For cell translocating AMPs, it can be sub-divided into AMPs inducing ROS, DNA damage, transcription inhibition, mitochondrial function disruption and DNA synthesis inhibition (Muthirulan & Chandrasekaran, 2016), Fig. 2 displays the strategy for discriminate membrane acting and cell-ranslocating AMP. Atomic force microscopy is a tool of great power to depict the dynamics of biologically relevant processes at aqueous interfaces with high spatial resolution.
The rapid increase of drug resistance in conventional antibiotics promotes the discovery and development of antimicrobial peptides (AMPs), which are important components of natural immunity. Due to the specific action modes targeting for cell membrane, AMPs have become a promising substance for bacterium inhibitions barely with no drug resistance. Here we reviewed the structure characteristics related with bactericide actions, including peptides constituents, molecular length, molecular charges and so on. And we also summarized the common mode of actions of AMPs raised by researchers. The structure-mechanism relationship was discussed. The commonly used methods for mechanism study were also listed.
The structures of AMPs were strongly related with its efficacy, such as constituents, molecular length, charges, secondary structures. The main mechanisms of AMPs were divided into membrane targets and intracellular targets. Some advanced and new techniques were applied in the mechanism study. Most of the existed mechanisms are still suppositional, and techniques were mainly focused on membrane-associated mechanism, only few of them were related with intracellular factors. Therefore, more new methods are expected to explore the authentic mechanisms and better results as well as more intracellular targets. This review provides a reference for future AMPs chemical modification and mechanism exploration.
Gene expressions and biological activities of two novel antimicrobial peptides (AMPs) isolated from leucocytes of the rock bream, Oplegnathus fasciatus
2018, Aquaculture
Citation Excerpt :
In a previous study, we also identified and reported on the molecular characterisation and biological activity of the moronecidin and piscidin in the rock bream, O. fasciatus (Bae et al., 2014, 2016). The existence of multiple isoforms of piscidins in fish suggests a putative association between the roles of the various piscidins in innate immunity and invading pathogens, particularly in environments that are rich in microbial pathogens (Marcellini et al., 2009; Shai, 2009; Muthirulan and Chandrasekaran, 2016). In this study, we identified additionally two novel piscidin isoforms, rock bream piscidin 6 and piscidin 7 (RP6 and RP7) and described their molecular characterisation.
Antimicrobial peptides (AMPs) process strong antimicrobial activity and play a major role in protect host against pathogens. Piscidins, one of main AMPs of fish, are considered as crucial effectors of innate immune response as well as modulators of adaptive immune response. In this study, we discovered two rock bream piscidin paralogs (RP6 and RP7) from leucocytes of the rock bream and investigated their gene expression patterns and antimicrobial and cytotoxic activities. Their open reading frames (ORF) consisted of 240 and 249 base pairs (bp), encoding 80 and 83 amino acid residues, respectively, and both conserved AMP 12 domain. The predicted tertiary structure of the AMP 12 domain of RP6 and RP7 is an amphipathic alpha-helix structure. RP6 and RP7 mRNAs are ubiquitously expressed in healthy fish, the most expression of both in the PBLs. RP6 expression was more induced after pathogen infection relative to that of RP7, while RP7 exhibited stronger antimicrobial and haemolytic activity compared with RP6. Ultimately, the powerful antibacterial effect of RP7 would allow a small amount of RP7 to protect the host from pathogens and RP6 may have other immune response function in rock bream in addition to direct pathogen killing, suggesting that RP6 and RP7 have important function in rock bream immune system. Also, the studies on these RP6 and RP7 will be useful as important data on the substitution of antimicrobials in aquaculture of rock bream.
Antimicrobial Peptides and Macromolecules for Combating Microbial Infections: From Agents to Interfaces
2022, ACS Applied Bio Materials

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Technical noteMicrobial flow cytometry: An ideal tool for prospective antimicrobial drug development

Abstract

Drug Discov. Today

Biophys. J.

Peptides

Methods

Cathelicidins, multifunctional peptides of the innate immunity

J. Leukoc. Biol.

Antimicrobial peptides: new drugs for bad bugs?

Expert Opin. Biol. Ther.

Antimicrobial peptides: versatile biological properties

Int. J. Pept.

APD3: the antimicrobial peptide database as a tool for research and education

Nucleic Acids Res.

Database-guided discovery of potent peptides to combat HIV-1 or superbugs

Pharmaceuticals

Prediction of antimicrobial peptides based on sequence alignment and feature selection methods

PLoS One