Evaluation of chrono-amperometric signal detection for the analysis of genotoxicity by a whole cell biosensor

Buchinger, Sebastian; Grill, Pia; Morosow, Valeri; Ben‐Yoav, Hadar; Shacham‐Diamand, Yosi; Biran, Alva; Pedahzur, Rami; Belkin, Shimshon; Reifferscheid, Georg

doi:10.1016/j.aca.2009.11.027

Cited by 16 publications

(9 citation statements)

References 37 publications

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“…Buchinger et al . proposed the umu test based on chronoamperometric detection using the screen printed electrode, and the Michaelis-Menten constant was determined to be 1.78 mM [39]. As for this study, the obtained Michaelis-Menten constant was 0.68 mM at the 3,000 rpm.…”

Section: Resultsmentioning

confidence: 72%

“…It is applicable to optically opaque media and allows detection of a small sample volume without loss of sensitivity, unlike optical detection techniques such as UV/vis spectrometry and fluorometric detection. In some recent works, the benefits of electrochemical detection have been employed into genotoxicity assays based on the umu test [38,39]. For these assays, p -aminophenyl-β-D-galactopyranoside (PAPG) is used as a substrate to evaluate the enzymatic reaction.…”

Section: Introductionmentioning

confidence: 99%

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Electrochemical Genotoxicity Assay Based on a SOS/umu Test Using Hydrodynamic Voltammetry in a Droplet

Kuramitz

Sazawa

Nanayama

et al. 2012

Sensors

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The SOS/umu genotoxicity assay evaluates the primary DNA damage caused by chemicals from the β-galactosidase activity of S. typhimurium. One of the weaknesses of the common umu test system based on spectrophotometric detection is that it is unable to measure samples containing a high concentration of colored dissolved organic matters, sediment, and suspended solids. However, umu tests with electrochemical detection techniques prove to be a better strategy because it causes less interference, enables the analysis of turbid samples and allows detection even in small volumes without loss of sensitivity. Based on this understanding, we aim to develop a new umu test system with hydrodynamic chronoamperometry using a rotating disk electrode (RDE) in a microliter droplet. PAPG when used as a substrate is not electroactive at the potential at which PAP is oxidized to p-quinone imine (PQI), so the current response of chronoamperometry resulting from the oxidation of PAP to PQI is directly proportional to the enzymatic activity of S. typhimurium. This was achieved by performing genotoxicity tests for 2-(2-furyl)-3-(5-nitro-2-furyl)-acrylamide (AF-2) and 2-aminoanthracene (2-AA) as model genotoxic compounds. The results obtained in this study indicated that the signal detection in the genotoxicity assay based on hydrodynamic voltammetry was less influenced by the presence of colored components and sediment particles in the samples when compared to the usual colorimetric signal detection. The influence caused by the presence of humic acids (HAs) and artificial sediment on the genotoxic property of selected model compounds such as 4-nitroquinoline-N-oxide (4-NQO), 3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone (MX), 1,8-dinitropyrene (1,8-DNP) and 1-nitropyrene (1-NP) were also investigated. The results showed that the genotoxicity of 1-NP and MX changed in the presence of 10 mg·L−1 HAs. The genotoxicity of tested chemicals with a high hydrophobicity such as 1,8-DNP and 1-NP were decreased substantially with the presence of 1 g·L−1 sediment. This was not observed in the case of genotoxins with a low log Kow value.

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Section: Resultsmentioning

confidence: 72%

Section: Introductionmentioning

confidence: 99%

Electrochemical Genotoxicity Assay Based on a SOS/umu Test Using Hydrodynamic Voltammetry in a Droplet

Kuramitz

Sazawa

Nanayama

et al. 2012

Sensors

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“…For example, P. putida has been used for the detection of aromatic hydrocarbons and organophosphate nerve agents [24], while S. cerevisiae Y190 has been used for determining endocrine disruptor compounds [25]. In the same context, amperometric measurements have been used with E.coli, Salmonella typhimurium and Ralstonia eutropha in genotoxicity assays [26][27][28]. Clearly, using mammalian cells has considerable advantages over precaryotic ones, especially allowing for a more realistic reflection of the investigated toxic effects on biologic species of interest, including humans.…”

Section: Discussionmentioning

confidence: 99%

Cellular bioelectric profiling: a novel method to differentially assess the in vitro toxicity of pesticides

2017

JCEBC

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We present a novel approach for assessing pesticide toxicity on mammalian cells, by measuring the response patterns of four cell lines (two neuroblastoma and two fibroblast) to three different pesticide groups (carbamates, organophosphates, pyrethroids) at a broad range of concentrations. Two different aspects of the cellular bioelectric response are measured, namely the potential and the combined resistance + capacitance of the cell suspension. Both the cell line and the mode of measurement affected the observed in vitro cellular responses, with each cell type providing a unique pattern. The measured of resistance + capacitance through amperometry provided more reproducible results than potentiometry. The results of the study demonstrate the potential of bioelectric profiling as a tool for developing novel toxicity assays and associated biosensors with superior analytical capacity, speed of assay and the ability to respond to a broad spectrum of toxicants, at the same time satisfying the demand for increased cost-efficiency and ease of operation.

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“…The major modes of signal transduction and detection mechanisms of the electrochemical microbial sensors include amperometric, potentiometric, or conductometric mechanisms [39] (Figure 1). It is to be noted that most of the toxic analytes detected by microbial cell sensors can be classified under the broad categories of metal (including heavy metals) [40,41,42], naphthalene and salicylate [43,44,45], BTEX compounds (benzene, toluene, ethylbenzene, and xylenes) [9,46,47], polychlorinated biphenyl (PCB) [48], phenols [49,50], surfactants [51], aromatic hydrocarbons [12,52], genotoxic compounds [22,53,54], to name a few. These types of sensors are employed mostly in ecotoxicological studies involving water and environmental monitoring.…”

Section: Choice Of Biological Cells For Toxin Testingmentioning

confidence: 99%

Biotoxin Detection Using Cell-Based Sensors

Banerjee

Kintzios

Prabhakarpandian

2013

Toxins

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Cell-based biosensors (CBBs) utilize the principles of cell-based assays (CBAs) by employing living cells for detection of different analytes from environment, food, clinical, or other sources. For toxin detection, CBBs are emerging as unique alternatives to other analytical methods. The main advantage of using CBBs for probing biotoxins and toxic agents is that CBBs respond to the toxic exposures in the manner related to actual physiologic responses of the vulnerable subjects. The results obtained from CBBs are based on the toxin-cell interactions, and therefore, reveal functional information (such as mode of action, toxic potency, bioavailability, target tissue or organ, etc.) about the toxin. CBBs incorporate both prokaryotic (bacteria) and eukaryotic (yeast, invertebrate and vertebrate) cells. To create CBB devices, living cells are directly integrated onto the biosensor platform. The sensors report the cellular responses upon exposures to toxins and the resulting cellular signals are transduced by secondary transducers generating optical or electrical signals outputs followed by appropriate read-outs. Examples of the layout and operation of cellular biosensors for detection of selected biotoxins are summarized.

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Evaluation of chrono-amperometric signal detection for the analysis of genotoxicity by a whole cell biosensor

Cited by 16 publications

References 37 publications

Electrochemical Genotoxicity Assay Based on a SOS/umu Test Using Hydrodynamic Voltammetry in a Droplet

Electrochemical Genotoxicity Assay Based on a SOS/umu Test Using Hydrodynamic Voltammetry in a Droplet

Cellular bioelectric profiling: a novel method to differentially assess the in vitro toxicity of pesticides

Biotoxin Detection Using Cell-Based Sensors

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