scholarly journals Physics Comes to the Aid of Medicine—Clinically-Relevant Microorganisms through the Eyes of Atomic Force Microscope

Pathogens ◽  
2020 ◽  
Vol 9 (11) ◽  
pp. 969
Author(s):  
Mateusz Cieśluk ◽  
Piotr Deptuła ◽  
Ewelina Piktel ◽  
Krzysztof Fiedoruk ◽  
Łukasz Suprewicz ◽  
...  

Despite the hope that was raised with the implementation of antibiotics to the treatment of infections in medical practice, the initial enthusiasm has substantially faded due to increasing drug resistance in pathogenic microorganisms. Therefore, there is a need for novel analytical and diagnostic methods in order to extend our knowledge regarding the mode of action of the conventional and novel antimicrobial agents from a perspective of single microbial cells as well as their communities growing in infected sites, i.e., biofilms. In recent years, atomic force microscopy (AFM) has been mostly used to study different aspects of the pathophysiology of noninfectious conditions with attempts to characterize morphological and rheological properties of tissues, individual mammalian cells as well as their organelles and extracellular matrix, and cells’ mechanical changes upon exposure to different stimuli. At the same time, an ever-growing number of studies have demonstrated AFM as a valuable approach in studying microorganisms in regard to changes in their morphology and nanomechanical properties, e.g., stiffness in response to antimicrobial treatment or interaction with a substrate as well as the mechanisms behind their virulence. This review summarizes recent developments and the authors’ point of view on AFM-based evaluation of microorganisms’ response to applied antimicrobial treatment within a group of selected bacteria, fungi, and viruses. The AFM potential in development of modern diagnostic and therapeutic methods for combating of infections caused by drug-resistant bacterial strains is also discussed.

Coatings ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 121
Author(s):  
Ecaterina Matei ◽  
Carmen Gaidau ◽  
Maria Râpă ◽  
Laura Mihaela Stefan ◽  
Lia-Mara Ditu ◽  
...  

In this study, sodium alginate film (Alg) was coated with electrospun collagen glue (Col) extracted from rabbit skin waste, loaded with different commercial antimicrobial agents (chitosan, AG425K and ZnONPs) and investigated in terms of morphological, structural and biological properties. The coated nanostructures were characterized using scanning electron microscopy coupled with the energy-dispersive X-ray (SEM/EDS), Attenuated Total Reflectance Fourier-Transform Infrared spectroscopy (ATR FT-IR), and Atomic Force Microscopy (AFM) tests. The cytotoxicity was investigated on murine L929 fibroblasts using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide salt (MTT) and lactate dehydrogenase (LDH) assays. Microbiological tests were performed against Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853 and Candida albicans ATCC 27853 standard strains. In vitro cell culture tests showed a good cytocompatibility of the coated nanostructured systems, except the sample loaded with ZnONPs, which exhibited a highly cytotoxic effect. Alg-Col-ZnONPs nanostructure inhibited the growth and multiplication of the Staphylococcus aureus ATCC 25923 and Escherichia coli ATCC 25922 bacterial strains. The results of new coated nanostructures may be useful for the development of sustainable biomaterials in a circular economy, with bioactive properties for medical wound dressings.


2013 ◽  
Vol 56 (9) ◽  
pp. 811-817 ◽  
Author(s):  
Mi Li ◽  
LianQing Liu ◽  
Ning Xi ◽  
YueChao Wang ◽  
ZaiLi Dong ◽  
...  

2021 ◽  
Author(s):  
Iris K Lee ◽  
Daniel A Jacome ◽  
Joshua K Cho ◽  
Vincent Tu ◽  
Anthony Young ◽  
...  

Recently, several molecular imaging strategies have developed to image bacterial infections in humans. Nuclear approaches, specifically positron emission tomography (PET), affords sensitive detection and the ability to non-invasively locate infections deep within the body. Two key radiotracer classes have arisen: metabolic approaches targeting bacterial specific biochemical transformations, and antibiotic-based approaches that have inherent selectivity for bacteria over mammalian cells. A critical question for clinical application of antibiotic radiotracers is whether resistance to the template antibiotic abrogates specific uptake, thus diminishing the predictive value of the diagnostic test. We recently developed small-molecule PET radiotracers based on the antibiotic trimethoprim (TMP), including [11C]-TMP, and have shown their selectivity for imaging bacteria in preclinical models. Here, we measure the in vitro uptake of [11C]-TMP in pathogenic susceptible and drug-resistant bacterial strains. Both resistant and susceptible bacteria showed similar in vitro uptake, which led us to perform whole genome sequencing of these isolates to identify the mechanisms of TMP resistance that permit retained radiotracer binding. By interrogating these isolate genomes and a broad panel of previously sequenced strains, we reveal mechanisms where uptake or binding of TMP radiotracers can potentially be maintained despite the annotation of genes conferring antimicrobial resistance. Finally, we present several examples of patients with both TMP-sensitive and drug-resistant infections in our first-in-human experience with [11C]-TMP. This work underscores the ability of an antibiotic radiotracer to image bacterial infection in patients, which may allow insights into human bacterial pathogenesis, infection diagnosis, and antimicrobial response monitoring.


2021 ◽  
Vol 12 ◽  
Author(s):  
Claudia Müller ◽  
Caillan Crowe-McAuliffe ◽  
Daniel N. Wilson

Ribosomes that become stalled on truncated or damaged mRNAs during protein synthesis must be rescued for the cell to survive. Bacteria have evolved a diverse array of rescue pathways to remove the stalled ribosomes from the aberrant mRNA and return them to the free pool of actively translating ribosomes. In addition, some of these pathways target the damaged mRNA and the incomplete nascent polypeptide chain for degradation. This review highlights the recent developments in our mechanistic understanding of bacterial ribosomal rescue systems, including drop-off, trans-translation mediated by transfer-messenger RNA and small protein B, ribosome rescue by the alternative rescue factors ArfA and ArfB, as well as Bacillus ribosome rescue factor A, an additional rescue system found in some Gram-positive bacteria, such as Bacillus subtilis. Finally, we discuss the recent findings of ribosome-associated quality control in particular bacterial lineages mediated by RqcH and RqcP. The importance of rescue pathways for bacterial survival suggests they may represent novel targets for the development of new antimicrobial agents against multi-drug resistant pathogenic bacteria.


2019 ◽  
Author(s):  
Peter Spanogiannopoulos ◽  
Patrick H. Bradley ◽  
Jonathan Melamed ◽  
Ysabella Noelle Amora Malig ◽  
Kathy N. Lam ◽  
...  

Microbiome surveys indicate that pharmaceuticals are the top predictor of inter-individual variations in gut microbial community structure1, consistent with in vitro evidence that non-antibiotic (i.e. host-targeted) drugs inhibit gut bacterial growth2and are subject to extensive metabolism by the gut microbiome3,4. In oncology, bacterial metabolism has been implicated in both drug efficacy5,6and toxicity7,8; however, the degree to which bacterial sensitivity and metabolism can be driven by conserved pathways also found in mammalian cells remains poorly understood. Here, we show that anticancer fluoropyrimidine drugs broadly inhibit the growth of diverse gut bacterial strains. Media supplementation, transcriptional profiling (RNA-seq), and bacterial genetics implicated pyrimidine metabolism as a key target in bacteria, as in mammalian cells. Drug resistant bacteria metabolized 5FU to its inactive metabolite dihydrofluorouracil (DHFU) mimicking the major host pathway for drug clearance. Functional orthologs of the bacterial operon responsible (preTA) are widespread across human gut bacteria from the Firmicutes and Proteobacteria phyla. The observed conservation of both the targets and pathways for metabolism of therapeutics across domains highlights the need to distinguish the relative contributions of human and microbial cells to drug disposition9, efficacy, and side effect profiles.


2013 ◽  
Vol 2013 ◽  
pp. 1-10 ◽  
Author(s):  
Emilia Tomaszewska ◽  
Katarzyna Soliwoda ◽  
Kinga Kadziola ◽  
Beata Tkacz-Szczesna ◽  
Grzegorz Celichowski ◽  
...  

Dynamic light scattering is a method that depends on the interaction of light with particles. This method can be used for measurements of narrow particle size distributions especially in the range of 2–500 nm. Sample polydispersity can distort the results, and we could not see the real populations of particles because big particles presented in the sample can screen smaller ones. Although the theory and mathematical basics of DLS technique are already well known, little has been done to determine its limits experimentally. The size and size distribution of artificially prepared polydisperse silver nanoparticles (NPs) colloids were studied using dynamic light scattering (DLS) and ultraviolet-visible (UV-Vis) spectroscopy. Polydisperse colloids were prepared based on the mixture of chemically synthesized monodisperse colloids well characterized by atomic force microscopy (AFM), transmission electron microscopy (TEM), DLS, and UV-Vis spectroscopy. Analysis of the DLS results obtained for polydisperse colloids reveals that several percent of the volume content of bigger NPs could screen completely the presence of smaller ones. The presented results could be extremely important from nanoparticles metrology point of view and should help to understand experimental data especially for the one who works with DLS and/or UV-Vis only.


2020 ◽  
Vol 12 (19) ◽  
pp. 1709-1727 ◽  
Author(s):  
Yuan-Yuan Hu ◽  
Juan Wang ◽  
Tie-Jun Li ◽  
Rammohan R Yadav Bheemanaboina ◽  
Mohammad Fawad Ansari ◽  
...  

Aim: With the increasing emergence of drug-resistant bacteria, the need for new antimicrobial agents has become extremely urgent. This work was to develop sulfonyl thiazoles as potential antibacterial agents. Results & methodology: Novel hybrids of sulfonyl thiazoles were developed from commercial acetanilide and acetylthiazole. Hybrids 6e and 6f displayed excellent inhibitory efficacy against clinical methicillin-resistant Staphylococcus aureus (MRSA) (minimum inhibitory concentration = 1 μg/ml) without obvious toxicity toward normal mammalian cells (RAW 264.7). The combination uses were found to improve the antimicrobial ability. Further preliminary antibacterial mechanism experiments showed that the active molecule 6f could effectively interfere with MRSA membrane and insert into MRSA DNA. Conclusion: Compounds 6e and 6f could serve as potential DNA-targeting templates toward the development of promising antimicrobial agents.


2005 ◽  
Vol 11 (S03) ◽  
pp. 78-81
Author(s):  
R. P. Santos ◽  
L. M. Rebelo ◽  
E. F. Costa ◽  
A. A. X. Santiago ◽  
V. N. Freire ◽  
...  

Pollens appear like a fine to coarse powder that is liberated by the microsporangia of Gimnosperms and Angiosperms. The pollen grain wall, the sporoderm, envelopes the microgametophytes (male gametophytes), which produce the male gametes of seed plants. Pollen grains are interesting from the material science point of view since the native polymer, the sporopollenin, found in the sporoderm outer layer (exine), is one of the toughest known materials which is degraded by oxidation but is resistant to reduction. This property permits the sporopollenin persistence as an unaltered polymer in sediments of great age, e.g the Ordovician period, 400 million years ago. Sporopollenin is a mixture of fatty acids, phenyl-derivatives as p-coumaric acid, and carotenes [1]. Its nanostructure is not yet completed revealed. Therefore, more studies must be performed. A number of models have been proposed for the sporopollenin nanostructure of spores and pollen grains [2]. Rowley et al. [3-4] interpret exine structure as being formed by helical subunits, based on transmission and scanning electron microscope (TEM and SEM) studies. The atomic force microscopy (AFM) is the ideal method to study the sporopollenin nanostructure [5] since the arrangement of components is not visualized easily through other microscope techniques (e.g. TEM and SEM). In the present work, we used AFM to study the sporopollenin nanostructure of the Ilex paraguariensis A.St.Hil. exine, an Angiosperm (Aquifoliaceae).


1994 ◽  
Vol 332 ◽  
Author(s):  
R. Giles ◽  
S. Manne ◽  
C.M. Zaremba ◽  
A. Belcher ◽  
S. Mann ◽  
...  

ABSTRACTAfter describing some recent developments in atomic force microscopy (AFM), a specific application to the study of shell ultrastructure is examined in detail. By embedding bleached nacreous tablets in epoxy and imaging them with the atomic force microscope (AFM) during in situ dissolution, it was possible to visualize the topography of both the top faces of the tablets and the impressions in epoxy made by the bottom faces of the tablets. This epoxy imprint reproduced tablet features down to the 10 nm scale. Using this technique it should be possible to measure correspondence between topographic features on the proximal and distal faces of tablets, which is necessary to form a three-dimensional picture of the nacreous region. In addition to these dissolution experiments, growth experiments (in modified sea water) on bleached, embedded tablets indicated that aragonite grows on a tablet as asperities oriented along the c axis, normal to the tablet surface. No change was seen on the surface of the epoxy, which confirmed that the crystals were growing on the tablet surface, not spontaneously nucleating out of solution.


Sign in / Sign up

Export Citation Format

Share Document