Induction of Different Sensitization Patterns of MRSA to Antibiotics Using Electroporation

Novickij, Vitalij; Švedienė, Jurgita; Paškevičius, Algimantas; Markovskaja, Svetlana; Lastauskienė, Eglė; Zinkevičienė, Auksė; Girkontaitė, Irutė; Novickij, Jurij

doi:10.3390/molecules23071799

Cited by 10 publications

(13 citation statements)

References 44 publications

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“…As it can be seen in Figure 3-left, electroporation can be effectively used for protein extraction; moreover, different electroporation parameters affect the protein yield. As expected, electroporation protocol 32 × 100 µs, 20 kV/cm, 1 Hz most strongly affected bacterial viability (Figure 3-right) due to higher energy input (379 kJ/L) (Novickij et al, 2018). Nevertheless protein yields were lower compared to electroporation protocol where lower energy of the pulses was applied -65 kJ/L (8 × 100 µs, 20 kV/cm, 1 kHz), which suggests that the risen temperature during the treatment possibly damaged proteins.…”

Section: Comparison Of Extraction Of Proteins By Means Of Electroporasupporting

confidence: 75%

Evaluation and Optimization of Protein Extraction From E. coli by Electroporation

Meglič

Janež

Peterka

et al. 2020

Front. Bioeng. Biotechnol.

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Growing diversity of protein-based technologies dictates further development of bio manufacturing to lower the cost of production and maximize yields. Intracellularly expressed recombinant proteins must be extracted from production host prior to purification. Use of electroporation to obtain proteins from bacteria and yeasts has been demonstrated in several studies for different modes of operation and formats. Here we tested various protocols for protein extraction from Escherichia coli by means of electroporation. The tested protocols were compared to established extraction methods of ultrasonication and glass-bead milling in terms of protein yields and content of impurities such as host cell DNA and endotoxins in the lysate. Protein extraction yield was maximal when exponentially growing bacteria were treated at 37 • C, regardless of the electroporation mode of operation (batch or flow). We were unable to eliminate co-extraction of host DNA and endotoxins, but with 8 × 1 ms, 5 kV/cm, 1 Hz pulses they were minimized. Yields with optimized electroporation (up to 86 g protein/kg dry weight) were inferior to those in ultrasonication (up to 144 g protein/kg dry weight) and glass-bead milling (up to 280 g protein/kg dry weight). Nevertheless, electroporation largely avoids cell lysis and disintegration with which the extract is a mix of extracted proteins with debris of the bacterial envelope and bacterial DNA, which necessitates further purification.

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Section: Comparison Of Extraction Of Proteins By Means Of Electroporasupporting

confidence: 75%

Evaluation and Optimization of Protein Extraction From E. coli by Electroporation

Meglič

Janež

Peterka

et al. 2020

Front. Bioeng. Biotechnol.

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“…It seems that the reason for this phenomenon is the ability of amphotericin B [46] and aminopenicillins [47] to interfere with the integrity of the cellular structures and a disintegration-dependent increase in uptake of 3-BP into microbial cells. An analogous mechanism of sensitizing bacterial cells to antibiotics has been proven against methicillin-resistant S. aureus exposed to electroporation [48]. In the Visca et al [26] study, the existence of an additive interaction of 3-BP with TET and chloramphenicol against S. aureus has also been demonstrated.…”

Section: Discussionmentioning

confidence: 96%

In Vitro Activity of 3-Bromopyruvate, an Anticancer Compound, Against Antibiotic-Susceptible and Antibiotic-Resistant Helicobacter pylori Strains

Krzyżek

Franiczek

Krzyżanowska

et al. 2019

Cancers

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Helicobacter pylori (H. pylori) is a bacterium capable of inducing chronic active gastritis, which in some people, develops into gastric cancers. One of the substances that may be useful in the eradication of this microorganism is 3-Bromopyruvate (3-BP), an anticancer compound with antimicrobial properties. The aim of this article was to determine the activity of 3-BP against antibiotic-susceptible and antibiotic-resistant H. pylori strains. The antimicrobial activity was determined using a disk-diffusion method, broth microdilution method, time-killing assay, and checkerboard assay. The research was extended by observations using light, fluorescence, and scanning electron microscopy. The growth inhibition zones produced by 2 mg/disk with 3-BP counted for 16–32.5 mm. The minimal inhibitory concentrations (MICs) ranged from 32 to 128 μg/mL, while the minimal bactericidal concentrations (MBCs) for all tested strains had values of 128 μg/mL. The time-killing assay demonstrated the concentration-dependent and time-dependent bactericidal activity of 3-BP. The decrease in culturability below the detection threshold (<100 CFU/mL) was demonstrated after 6 h, 4 h, and 2 h of incubation for MIC, 2× MIC, and 4× MIC, respectively. Bacteria treated with 3-BP had a several times reduced mean green/red fluorescence ratio compared to the control samples, suggesting bactericidal activity, which was independent from an induction of coccoid forms. The checkerboard assay showed the existence of a synergistic/additive interaction of 3-BP with amoxicillin, tetracycline, and clarithromycin. Based on the presented results, it is suggested that 3-BP may be an interesting anti-H. pylori compound.

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“…Also, where stated in these previous studies, the post-pulse incubation times before diluting out of the antibiotics have been as long as 24 h ( Novickij et al, 2018a ; Kuyukina et al, 2020 ; Martens et al, 2020 ). Implementation of such a long incubation for on-site hospital wastewater treatment would require a reservoir in which the undiluted antibiotic concentration would be retained until release into the communal sewage (and thus dilution), and the required reservoir volume would likely be prohibitive.…”

Section: Introductionmentioning

confidence: 96%

“…To date, the majority of studies that have combined electroporation and antibacterials [see Table 2 in Garner (2019) ] have focused on substances permissible in food and beverage processing. Still, five recent studies have quantified the potentiation of inactivation rates for combination of electroporation and clinical antibiotics ( Korem et al, 2018 ; Novickij et al, 2018a ; Vadlamani et al, 2018 , 2020 ; Rubin et al, 2019 ), with their exposure parameters and results summarized in Table 1 . Two further studies ( Kuyukina et al, 2020 ; Martens et al, 2020 ) used the disk diffusion test to study electroporation-induced increase in susceptibility of Rhodococcus ruber to five and Escherichia coli to four different antibiotics, at increasing concentrations, and confirmed that such susceptibility increases are generally achievable.…”

Section: Introductionmentioning

confidence: 99%

Electroporation as an Efficacy Potentiator for Antibiotics With Different Target Sites

2021

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Antibiotic resistance is a global health threat, and there is ample motivation for development of novel antibacterial approaches combining multiple strategies. Electroporation is among the promising complementary techniques – highly optimizable, effective against a broad range of bacteria, and largely impervious to development of resistance. To date, most studies investigating electroporation as an efficacy potentiator for antibacterials used substances permissible in food industry, and only few used clinical antibiotics, as acceptable applications are largely limited to treatment of wastewaters inherently contaminated with such antibiotics. Moreover, most studies have focused mainly on maximal achievable effect, and less on underlying mechanisms. Here, we compare Escherichia coli inactivation potentiation rates for three antibiotics with different modes of action: ampicillin (inhibits cell wall synthesis), ciprofloxacin (inhibits DNA replication), and tetracycline (inhibits protein synthesis). We used concentrations for each antibiotic from 0 to 30× its minimum inhibitory concentration, a single 1-ms electric pulse with amplitude from 0 to 20 kV/cm, and post-pulse pre-dilution incubation either absent (≲1 min) or lasting 60 min, 160 min, or 24 h. Our data show that with incubation, potentiation is significant for all three antibiotics, increases consistently with pulse amplitude, and generally also with antibiotic concentration and incubation time. With incubation, potentiation for ampicillin was rather consistently (although with weak statistical significance) superior to both ciprofloxacin and tetracycline: ampicillin was superior to both in 42 of 48 data points, including 7 with significance with respect to both, while at 60- and 160-min incubation, it was superior in 31 of 32 data points, including 6 with significance with respect to both. This suggests that electroporation potentiates wall-targeting antibiotics more than those with intracellular targets, providing motivation for in-depth studies of the relationship between the mode of action of an antibiotic and its potentiation by electroporation. Identification of substances permissible in foods and targeting the cell wall of both Gram-negative and Gram-positive bacteria might provide candidate antibacterials for broad and strong potentiation by electroporation applicable also for food preservation.

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Induction of Different Sensitization Patterns of MRSA to Antibiotics Using Electroporation

Cited by 10 publications

References 44 publications

Evaluation and Optimization of Protein Extraction From E. coli by Electroporation

Evaluation and Optimization of Protein Extraction From E. coli by Electroporation

In Vitro Activity of 3-Bromopyruvate, an Anticancer Compound, Against Antibiotic-Susceptible and Antibiotic-Resistant Helicobacter pylori Strains

Electroporation as an Efficacy Potentiator for Antibiotics With Different Target Sites

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