Previous studies clearly indicated that membrane enzymes - ATPases i.e. Na, K-ATPase and Mg-ATPase, responded to presence of various organic and inorganic pollutants. In this work effect of mercury and cadmium on these enzymes activities was investigated in synaptic plasma membranes partially immobilized on microliter plate. Comparing those activities with the control enzyme activities obtained with native and partially immobilized mambranes it was concluded that: a. both metals exerted a concentration-dependent inhibition of investigated enzymes, b. for partially immobilized membranes estimated half maximum inhibition (IC50) values for Na, K-ATPase were IC50 (Hg) = 0.9 micromol/cubic cm, IC50 (cd) = 35 micromol/cubic cm and for Mg - ATPase IC50 (Hg) = 3.5 micromol/cubic cm, IC50 (Cd) = 36 micromol/cubic cm; for native membranes IC50 for Na,K-ATPase were IC50 (Hg) =3.3 micromol/cubic cm, IC50 (Cd) = 2 micromol/cubic cm and for Mg ATPase IC50 (Hg) = 2.3 micromol/cubic cm, IC50 (Cd) = 0.2 mmol/cubic cm. Obtained results indicate avaibility of microtitar plates for partially immobilization of membranes with aim to form a new biosensor for heavy metals detection.

Possibilities of application of transmembrane enzymes as a biological component of a biosensor for water quality control and detection of toxical substances were performed. Synaptic plasma membranes (SPMs) were adsorbed on nitrocellulose filters. The adsorption of SPMs was followed by determination of the transmembrane enzyme Na,K-ATPase. The optimal conditions for SPM adsorption on nitrocellulose filters were determined: 25 microgram per nitrocellulose filter disc during 1 hour of incubation, on - 20 deg C. The ATPase activity of adsorbed SPM showed, that almost 30% of enzymic activity was detected on nitrocellulose filters in mentioned conditions. This results showed that adsorption of SPM on solid support enhancing enzymatic stability and enable its industrial and analytical application.

In our earlier work, we have shown that enzymes from rat brain synaptosomal membranes, Na(+)/K(+)-ATPase and Mg(2+)-ATPase, are promising biological components of a biosensor for lead detection. In this work, we represent our results of investigation with the same enzymes as biological components for the biosensor in presence of Hg(2+) ions in water. It was established that IC50 for Na(+)/K(+)-ATPase and Mg(2+)-ATPase is 6.9 and 5.5 x 10E-6 M, and the percentages of inhibition are 96% and 77% respectively. We concluded that these enzymes could be the base for developing biosensors for the presence of Hg(2+) ion in water. Since these enzymes maintain a stable activity for a longer period of time, they could be appropriate as components of biosensors for monitoring water quality.

Spectrophotometric method for determination of inorganic phosphate liberated in hydrolysis of ATP catalyzed by ATPase was modified in order to obtain faster procedure, which could also be used in none laboratory conditions. The modification has some advantages compared to the most used Pennial method: a) the reagents are stable for several months; b) the 45 min procedure of phosphomolibdate extraction by isobutanole-benzene is ommited, and the method is not dangerous for the analyst; c) color develops after 20 min. The method was tested on the determination of inorganic phosphate in the presence of cadmium nitrate as inhibitor of ATPase activity. The results were compared to the results obtained by Pennial method. The results obtained have shown some good agreements.

Activities of rat brain synaptic plasma membrane (SPM)Na(+)/K(+)-ATPase and Mg(2+)-ATPase were investigated by in vitro individual and simultaneous exposure to ions of the first transition series metals (Cu(2+), Zn(2+), Fe(2+)). All investigated metals produced a larger maximum inhibition of Na(+)/K(+)-ATPase than Mg(2+)-ATPase activity. Metal concentrations causing 50% inhibition of Na(+)/K(+)-ATPase activities were: Cu(2+) 7.1 microM Zn(2+) 19 microM Fe(2+) 25 microM. Simulataneous exposure to metal combinations: Cu(2+)/Zn(2+), Cu(2+)/Fe(2+) and Zn(2+)/Fe(2+) inhibited both the Na(+)/K(+)-ATPase and Mg(2+)-ATPase activity synergistically, i.e., more than the sum of the metal induced inhibitions assayed separately. It is also established that the plot of logarithm IC50 values of Na(+)/K(+)-ATPase activity vs. the ionic radius of (Cu(2+), Zn(2+), Fe(2+)) is a straight line that enables to predict the value IC50 for other metals of the first transition series. The results obtained show that Na(+)/K(+)-ATPase is promising as a biological component of biosensor.

Glyphosate (GLY) is a broad-spectrum herbicide used worldwide to control broadleaf sedge, and grass weeds to control non-specific vegetation. Although it was evaluated as non-toxic agent in 20ᵗʰ century, its carcinogenic and genotoxic potential has being intensively investigated all over the world in the last decade. Moreover, the combination of GLY and 2,4-dichlorophenoxyacetic acid (2,4-D) has been widely applied. Although genotoxicity of GLY has been evaluated in vivo studies, there is no report in the literature for the monitoring of in vitro biointeraction of GLY and double stranded DNA, or how effect the combination of GLY and 2,4-D onto DNA. Herein, an electrochemical biosensor platform was developed for detection of the pesticide-DNA interaction by using disposable pencil graphite electrodes (PGEs). First, voltammetric detection of the interaction between GLY and DNA was investigated and the electrochemical characterization of the interaction was achieved. Taking a step further, the synergistic genotoxic effect of the mixture of GLY and 2,4-dichlorophenoxyacetic acid (2,4-D) or the mixture of their herbicide forms onto DNA could be monitored. This effect was concentration dependent, and the herbicide of GLY or the use of mixture of herbicides of GLY and 2,4-D had more genotoxic effect than analytical grade of the active molecules, GLY and 2,4-D. The single-use PGEs provided to fabricate robust, eco-friendly and time saver recognition platform for monitoring of herbicide-DNA interaction with the sensitive and reliable results. It is expected that this study will lead to be designed miniaturized lab-on-a chip platforms for on-line analysis of the pesticide-nucleic acid interactions.

The ubiquity of pollutants, such as agrochemicals and heavy metals, constitute a serious risk to human health. To evaluate the induction of DNA damage and programmed cell death (PCD), root cells of Allium cepa and Vicia faba were treated with two organophosphate insecticides (OI), fenthion and malathion, and with two heavy metal (HM) salts, nickel nitrate and potassium dichromate. An alkaline variant of the comet assay was performed to identify DNA breaks; the results showed comets in a dose-dependent manner, while higher concentrations induced clouds following exposure to OIs and HMs. Similarly, treatments with higher concentrations of OIs and HMs were analyzed by immunocytochemistry, and several structural characteristics of PCD were observed, including chromatin condensation, cytoplasmic vacuolization, nuclear shrinkage, condensation of the protoplast away from the cell wall, and nuclei fragmentation with apoptotic-like corpse formation. Abiotic stress also caused other features associated with PCD, such as an increase of active caspase-3-like protein, changes in the location of cytochrome C (Cyt C) toward the cytoplasm, and decreases in extracellular signal-regulated protein kinase (ERK) expression. Genotoxicity results setting out an oxidative via of DNA damage and evidence the role of the high affinity of HM and OI by DNA molecule as underlying cause of genotoxic effect. The PCD features observed in root cells of A. cepa and V. faba suggest that PCD takes place through a process that involves ERK inactivation, culminating in Cyt C release and caspase-3-like activation. The sensitivity of both plant models to abiotic stress was clearly demonstrated, validating their role as good biosensors of DNA breakage and PCD induced by environmental stressors.

Polycyclic Aromatic Hydrocarbons (PAHs) are among the most ubiquitous environmental pollutants of high global concern. PAHs belong to a diverse family of hydrocarbons with over one hundred compounds known, each containing at least two aromatic rings in their structure. Due to hydrophobic nature, PAHs tend to accumulate in the aquatic sediments, leading to bioaccumulation and elevated concentrations over time. In addition to their well-manifested mutagenic and carcinogenic effects in humans, they pose severe detrimental effects to aquatic life. The high eco-toxicity of PAHs has attracted a number of reviews, each dealing specifically with individual aspects of this global pollutant. However, efficient management of PAHs warrants a holistic approach that combines a thorough understanding of their physico-chemical properties, modes of environmental distribution and bioaccumulation, efficient detection, and bioremediation strategies. Currently, there is a lack of a comprehensive study that amalgamates all these aspects together. The current review, for the first time, overcomes this constraint, through providing a high level comprehensive understanding of the complexities faced during PAH management, while also recommending future directions through potentially viable solutions. Importantly, effective management of PAHs strongly relies upon reliable detection tools, which are currently non-existent, or at the very best inefficient, and therefore have a strong prospect of future development. Notably, the currently available biosensor technologies for PAH monitoring have not so far been compiled together, and therefore a significant focus of this article is on biosensor technologies that are critical for timely detection and efficient management of PAHs. This review is focussed on inland aquatic ecosystems with an emphasis on fish biodiversity, as fish remains a major source of food and livelihood for a large proportion of the global population. This thought provoking study is likely to instigate new collaborative approaches for protecting aquatic biodiversity from PAHs-induced eco-toxicity.

There are strong drivers to increasingly adopt bioremediation as an effective technique for risk reduction of hydrocarbon impacted soils. Researchers often rely solely on chemical data to assess bioremediation efficiently, without making use of the numerous biological techniques for assessing microbial performance. Where used, laboratory experiments must be effectively extrapolated to the field scale. The aim of this research was to test laboratory derived data and move to the field scale. In this research, the remediation of over thirty hydrocarbon sites was studied in the laboratory using a range of analytical techniques. At elevated concentrations, the rate of degradation was best described by respiration and the total hydrocarbon concentration in soil. The number of bacterial degraders and heterotrophs as well as quantification of the bioavailable fraction allowed an estimation of how bioremediation would progress. The response of microbial biosensors proved a useful predictor of bioremediation in the absence of other microbial data. Field-scale trials on average took three times as long to reach the same endpoint as the laboratory trial. It is essential that practitioners justify the nature and frequency of sampling when managing remediation projects and estimations can be made using laboratory derived data. The value of bioremediation will be realised when those that practice the technology can offer transparent lines of evidence to explain their decisions. Detailed biological, chemical and physical characterisation reduces uncertainty in predicting bioremediation.

Photoelectrocatalysis driven by visible light offers a new and potentially powerful technology for the remediation of water contaminated by organo-xenobiotics. In this study, the performance of a visible light-driven photoelectrocatalytic (PEC) batch reactor, applying a tungsten trioxide (WO3) photoelectrode, to degrade the model pollutant 2,4-dichlorophenol (2,4-DCP) was monitored both by toxicological assessment (biosensing) and chemical analysis. The bacterial biosensor used to assess the presence of toxicity of the parent molecule and its breakdown products was a multicopy plasmid lux-marked E. coli HB101 pUCD607. The bacterial biosensor traced the removal of 2,4-DCP, and in some case, its toxicity response suggests the identification of transient toxic intermediates. The loss of the parent molecule, 2,4-DCP determined by HPLC, corresponded to the recorded photocurrents. Photoelectrocatalysis offers considerable potential for the remediation of chlorinated hydrocarbons, and that the biosensor based toxicity results identified likely compatibility of this technology with conventional, biological wastewater treatment. Visible light-driven photoelectrocatalysis has potential as a remediation technology in wastewater treatment.