Cell damage detection using Escherichia coli reporter plasmids: fluorescent and colorimetric assays

The metabolism of spoilage organism in the seafood is a major cause of fish spoilage. Lipid oxidation leads to production of intermediate peroxides causing bad odor and offflavour producing compounds. Hence, based on the hurdle technology method and applications of nanotechnology, the inhibitory effects of copper oxide nanoparticles coupled with vitamin-E (CuONPs+VE) was investigated with the aim of controlling the oxidative cell damage in seafood by estimating Reacting Oxygen Species (ROS) using 2’,7’- dichlorodihydrofluorescein diacetate (DCF) and preventing microbial spoilage by studying the cell viability using MTT assay in the present study. The MTT assay reveals that the CuONPs+VE inhibited the metabolic activity of spoilage bacteria which was confirmed from the inhibition percentage values against the test organisms. The CuONPs+VE exposed Escherichia coli reduced upto 86 ± 0.57% after enumerating the CFU in a Plate Count Agar media. Salmonella sp and Shigella sp exhibited 89 ± 1.04% and 85 ± 0.25% respectively. The qualitative inhibitory effect of Nanometal vitamin complex against three test bacteria, were evaluated by agar diffusion method. Among the four different concentrations of Nanometal vitamin complex, the higher concentration (4X – 40μg/ml) exhibited maximum inhibitory effects in terms of zone of clearance. Against Escherichia coli, 12.3 ± 1.25mm of inhibitory zones were found evident. Salmonella sp showed 15.6 ± 0.57mm of inhibitory zones and Shigella sp revealed the inhibition zone of about 16.6 ± 1.04mm. The anti-oxidative defence mechanisms operating in the cells for scavenging of ROS was performed using DCF. The cells which are pre-incubated with CuONPs+VE and then exposed to H2O2 show insignificant green emission indicating a clear advantage of the nanoparticle form of vitamin-E (CuONPs+VE) in preventing oxidative cell death.

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Effect of Gamma Radiation and Oregano Essential Oil on Murein and ATP Concentration of Escherichia coli O157:H7

Journal of Food Protection ◽

10.4315/0362-028x-68.12.2571 ◽

2005 ◽

Vol 68 (12) ◽

pp. 2571-2579 ◽

Cited By ~ 24

Author(s):

STÉPHANE CAILLET ◽

FRANÇOIS SHARECK ◽

MONIQUE LACROIX

Keyword(s):

Escherichia Coli ◽

Essential Oil ◽

Gamma Radiation ◽

Cell Damage ◽

Wall Structure ◽

High Dose ◽

Escherichia Coli O157 ◽

Oregano Essential Oil ◽

Intracellular Atp ◽

Oregano Oil

This study was carried out to evaluate the effects of gamma radiation alone or in combination with oregano essential oil on the murein composition of Escherichia coli O157:H7 and on the intracellular and extracellular concentrations of ATP. The bacterial strain was treated with three radiation doses: 0.4 kGy to induce cell damage, 1.1 kGy to obtain a viable but nonculturable state, and 1.3 kGy to cause cell death. Oregano essential oil was used at 0.006 and 0.025% (wt/vol), which is the MIC. All treatments had a significant effect (P ≤ 0.05) on the murein composition, although some muropeptides did not seem to be affected by the treatment. Each treatment had a different effect on the relative percentage and number of muropeptides. There was a significant correlation (P ≤ 0.05) between the decrease in intracellular ATP and the increase in extracellular ATP following treatment of the cells with oregano oil. The reduction of intracellular ATP was even more important when oregano oil was combined with irradiation, but irradiation alone at a high dose (≤1.1 kGy) significantly decreased (P ≤ 0.05) the internal ATP without affecting the external ATP. Transmission electron microscopic examination revealed that oregano oil and irradiation have an effect on cell wall structure.

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Comparison of intense pulsed light- and ultraviolet (UVC)-induced cell damage in Listeria monocytogenes and Escherichia coli O157:H7

Food Control ◽

10.1016/j.foodcont.2011.11.032 ◽

2012 ◽

Vol 25 (2) ◽

pp. 654-659 ◽

Cited By ~ 62

Author(s):

Chan-Ick Cheigh ◽

Mi-Hyun Park ◽

Myong-Soo Chung ◽

Jung-Kue Shin ◽

Young-Seo Park

Keyword(s):

Escherichia Coli ◽

Listeria Monocytogenes ◽

Cell Damage ◽

Intense Pulsed Light ◽

Escherichia Coli O157 ◽

Pulsed Light

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Autoinducer-2 of quorum sensing is involved in cell damage caused by avian pathogenic Escherichia coli

Microbial Pathogenesis ◽

10.1016/j.micpath.2016.08.033 ◽

2016 ◽

Vol 99 ◽

pp. 247-252 ◽

Cited By ~ 4

Author(s):

Zhen-Qiang Cui ◽

Zong-Mei Wu ◽

Yun-Xing Fu ◽

Dao-Xiu Xu ◽

Xun Guo ◽

...

Keyword(s):

Escherichia Coli ◽

Quorum Sensing ◽

Cell Damage ◽

Avian Pathogenic Escherichia Coli ◽

Autoinducer 2 ◽

Pathogenic Escherichia Coli

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Enhanced Adhesion and Cell Damage by Escherichia coli Harboring hly, papC and cnf-1genes to the Uroepithelium in Diabetic Mouse Bladder Model

Journal of Pure and Applied Microbiology ◽

10.22207/jpam.11.1.75 ◽

2017 ◽

Vol 11 (1) ◽

pp. 577-583

Author(s):

H Anandkumar ◽

CS Vinodkumar ◽

G Soham ◽

B Basavaraj ◽

Achut Rao ◽

...

Keyword(s):

Escherichia Coli ◽

Cell Damage ◽

Diabetic Mouse ◽

Bladder Model

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A comprehensive conceptual and computational dynamics framework for Autonomous Regeneration Systems

10.1101/820613 ◽

2019 ◽

Author(s):

Tran Nguyen Minh-Thai ◽

Sandhya Samarasinghe ◽

Michael Levin

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Damage Detection ◽

Cell Damage ◽

Entropy Change ◽

Cell Network ◽

Computational Dynamics ◽

Differentiated Cells ◽

Tissue Structures ◽

Pattern Information

AbstractThis paper presents a new conceptual and computational dynamics framework for damage detection and regeneration in multicellular structures similar to living animals. The model uniquely achieves complete and accurate regeneration from any damage anywhere in the system. We demonstrated the efficacy of the proposed framework on an artificial organism consisting of three tissue structures corresponding to the head, body and tail of a worm. Each structure consists of a stem cell surrounded by a tissue of differentiated cells. We represent a tissue as an Auto-Associative Neural Network (AANN) with local interactions and stem cells as a self-repair network with long-range interactions. We also propose another new concept, Information Field which is a mathematical abstraction over traditional components of tissues, to keep minimum pattern information of the tissue structures to be accessed by stem cells in extreme cases of damage. Through entropy, a measure of communication between a stem cell and differentiated cells, stem cells monitor the tissue pattern integrity, violation of which triggers damage detection and tissue repair. Stem cell network monitors its state and invokes stem cell repair in the case of stem cell damage. The model accomplishes regeneration at two levels: In the first level, damaged tissues with intact stem cells regenerate themselves. Here, stem cell identifies entropy change and finds the damage and regenerates the tissue in collaboration with the AANN. In the second level, involving missing whole tissues and stem cells, the remaining stem cell(s) access the information field to restore the stem cell network and regenerate missing tissues. In the case of partial tissue damage with missing stem cells, the two levels collaborate to accurately restore the stem cell network and tissues. This comprehensive hypothetical framework offers a new way to conceptualise regeneration for better understanding the regeneration processes in living systems. It could also be useful in biology for regenerative medicine and in engineering for building self-repairing biobots.

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Secreted autotransporter toxin (Sat) induces cell damage during enteroaggregative Escherichia coli infection

PLoS ONE ◽

10.1371/journal.pone.0228959 ◽

2020 ◽

Vol 15 (2) ◽

pp. e0228959 ◽

Cited By ~ 1

Author(s):

Paulo C. G. Vieira ◽

Abraham O. Espinoza-Culupú ◽

Roberto Nepomuceno ◽

Marina R. Alves ◽

Ivo Lebrun ◽

...

Keyword(s):

Escherichia Coli ◽

Cell Damage ◽

Escherichia Coli Infection

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Goat milk with and without increased concentrations of lysozyme improves repair of intestinal cell damage induced by enteroaggregative Escherichia coli

BMC Gastroenterology ◽

10.1186/1471-230x-12-106 ◽

2012 ◽

Vol 12 (1) ◽

Cited By ~ 9

Author(s):

Eunice B Carvalho ◽

Elizabeth A Maga ◽

Josiane S Quetz ◽

Ila FN Lima ◽

Hemerson YF Magalhães ◽

...

Keyword(s):

Escherichia Coli ◽

Cell Damage ◽

Goat Milk ◽

Intestinal Cell

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Inhibition of P2X Receptors Protects Human Monocytes against Damage by Leukotoxin from Aggregatibacter actinomycetemcomitans and α-Hemolysin from Escherichia coli

Infection and Immunity ◽

10.1128/iai.00674-16 ◽

2016 ◽

Vol 84 (11) ◽

pp. 3114-3130 ◽

Cited By ~ 12

Author(s):

Steen K. Fagerberg ◽

Martin R. Jakobsen ◽

Marianne Skals ◽

Helle A. Praetorius

Keyword(s):

Escherichia Coli ◽

Plasma Membranes ◽

Cell Damage ◽

Aggregatibacter Actinomycetemcomitans ◽

P2x Receptors ◽

Receptor Expression ◽

Human Monocytes ◽

P2 Receptor ◽

Content Type ◽

Tract Infections

α-Hemolysin (HlyA) from Escherichia coli and leukotoxin A (LtxA) from Aggregatibacter actinomycetemcomitans are important virulence factors in ascending urinary tract infections and aggressive periodontitis, respectively. The extracellular signaling molecule ATP is released immediately after insertion of the toxins into plasma membranes and, via P2X receptors, is essential for the erythrocyte damage inflicted by these toxins. Moreover, ATP signaling is required for the ensuing recognition and phagocytosis of damaged erythrocytes by the monocytic cell line THP-1. Here, we investigate how these toxins affect THP-1 monocyte function. We demonstrate that both toxins trigger early ATP release and a following increase in the intracellular Ca 2+ concentration ([Ca 2+ ] i ) in THP-1 monocytes. The HlyA- and LtxA-induced [Ca 2+ ] i response is diminished by the P2 receptor antagonist in a pattern that fits the functional P2 receptor expression in these cells. Both toxins are capable of lysing THP-1 cells, with LtxA being more aggressive. Either desensitization or blockage of P2X 1 , P2X 4 , or P2X 7 receptors markedly reduces toxin-induced cytolysis. This pattern is paralleled in freshly isolated human monocytes from healthy volunteers. Interestingly, only a minor fraction of the toxin-damaged THP-1 monocytes eventually lyse. P2X 7 receptor inhibition generally prevents cell damage, except from a distinct cell shrinkage that prevails in response to the toxins. Moreover, we find that preexposure to HlyA preserves the capacity of THP-1 monocytes to phagocytose damaged erythrocytes and may induce readiness to discriminate between damaged and healthy erythrocytes. These findings suggest a new pharmacological target for protecting monocytes during exposure to pore-forming cytolysins during infection or injury.

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