Aptamer based tools for environmental and therapeutic monitoring: A review of developments, applications, future perspectives

Nucleic acids in the form of aptamers play a growing and significant role in the targeted and rapid analysis of environmental sample composition and medical analyses. In this paper, the review of both aptamers synthesis methods as well as application of these short chain oligonucleotides (with critical comments on their strong and weak features) are given. The first ones include: systematic evolution of ligands by exponential enrichment (SELEX), high throughput aptamer identification screen (HAPIscreen), and a non-equilibrium capillary electrophoresis of equilibrium mixture (NECEEM). Afterwards, manuscript describes variety of sensors and biotests utilizing aptamers as active part of its action starting from electrochemical aptasensors, through optical to piezo-electric ones. Described biotests present basic developments in enzymelinked apta-sorbent assays (ELASA) that can be performed with different variations (enzyme-linked aptamer assay (ELAA), enzyme-linked oligonucleotide assay (ELONA) and aptamerlinked immobilized sorbent assay (ALISA)). Next, the review presents advantages and drawbacks of recent aptameric developments in versatile laboratory applications, namely medical ones, as well as analytical and bioassays. Utilitarian development of aptasensors and aptamers would strongly benefit from an assembly of interdisciplinary teams containing chemists, physicists, biologists, medical doctors, and material and electronic scientists, to determine the most effective application methodologies.

Ecotoxicological equilibria of triclosan inMicrotox, XenoScreen YES/YAS, Caco2, HEPG2 and liposomal systems are affected by the occurrence of other pharmaceutical and personal care emerging contaminants

M. Oliver, B. Kudłak , M. Wieczerzak , S. Reis, S. Lima, M. Segundo, M. Miro SCIENCE OF THE TOTAL ENVIRONMENT – 2020
Contaminants of emerging concernmay be considered as any chemicals or factorswhose unintended continuous release and persistence in the environmentmay lead to any observable undesirable response of living beings. Still not much is known on reciprocal toxicological impact of given chemicals when present in binary or more complex mixtures. In thiswork, an attemptwas thus undertaken to study the impact of butylparaben,methylparaben and diclofenac on toxicological behavior and properties of triclosan (at varying concentration levels)with respect to Microtox, XenoScreen YES/YAS, Caco-2, HEPG2, and liposomal systems.

Ultrasound assisted solvent extraction of porous membrane-packed samples followed by liquid chromatography-tandem mass spectrometry for determination of BADGE, BFDGE and their derivatives in packed vegetables

The problem of the presence of trace organic pollutants in food is of growing importance due to increasing awareness about their impact on newborns, infants and adults of reproductive age. Despite the fact that packaged food products offer many advantages, packaging can be a source of contamination for stored food. Thus, monitoring such pollution in food is of high importance. In this work, a novel methodology based on the solvent extraction of porous membrane-packed samples followed by liquid chromatography-tandem mass spectrometry was applied for the determination of bisphenol A diglycidyl ether (BADGE), bisphenol F diglycidyl ether (BFDGE) and their derivatives in packed vegetables. Several parameters of the extraction process were optimized, including the volume and type of extraction solvent as well as the sonication time. Due to advantages such as simplicity of use, short analysis time, and a reduction in the required amount solvent, the developed procedure can be considered green. In addition, the developed methodology was characterized by good validation parameters. Limit if quantitation (LOQ) was found to be in the range of 0.8 to 1.5 ng/g. The obtained recoveries varied from 78.3% to 111.2%. The repeatability of the extraction ranged between 0.6% and 5.8% (RSD). The proposed method was successfully applied to determine the presence of BADGE, BFDGE and their derivative compounds in the vegetable samples stored in different types of containers. The obtained data indicate that the majority of investigated samples were contaminated by chlorinated and hydroxyl derivatives of BADGE.

Assessment of the Bulgarian Wastewater Treatment Plants’ Impact on the Receiving Water Bodies

G. Yotova, S. Lazarova, B. Kudłak , B. Zlateva, V. Mihaylova, M. Wieczerzak , T. Venelinov, S. Tsakovski – MOLECULES – 2019
Deterioration of water quality is a major problem world widely according to many international non-governmental organizations (NGO). As one of the European Union (EU) countries, Bulgaria is also obliged by EU legislation to maintain best practices in assessing surface water quality and the efficiency of wastewater treatment processes. For these reasons studies were undertaken to utilize ecotoxicological (Microtox®, Phytotoxkit FTM, Daphtoxkit FTM), instrumental (to determine pH, electrical conductivity (EC), chemical oxygen demand, total suspended solids (TSS), total nitrogen (N) and phosphorus (P), chlorides, sulphates, Cr, Co, Cu, Cd, Ba, V, Mn, Fe, Ni, Zn, Se, Pb), as well as advanced chemometric methods (partial least squares–discriminant analysis (PLS-DA)) in data evaluation to comprehensively assess wastewater treatment plants' (WWTPs) effluents and surface waters quality around 21 major Bulgarian cities. The PLS-DA classification model for the physicochemical parameters gave excellent discrimination between WWTP effluents and surface waters with 93.65% correct predictions (with significant contribution of EC, TSS, P, N, Cl, Fe, Zn, and Se). The classification model based on ecotoxicological data identifies the plant test endpoints as having a greater impact on the classification model efficiency than bacterial, or crustaceans’ endpoints studied.

Bisphenols (A, S, and F) affect the basic hormonal activity determined for pharmaceuticals – Study of Saccharomyces cerevisiae

Pharmaceuticals and analogs of bisphenol A (BPA) are increasingly threatening environmental pollutants. In this study, mixtures of selected pharmaceuticals (diclofenac sodium salt, chloramphenicol, oxytetracycline hydrochloride, fluoxetine hydrochloride, estrone, ketoprofen, progesterone, gemfibrozil and androstenedione) were prepared with BPA and its two analogs (namely, bisphenols F and S) at such ratios to reflect environmentally detectable levels. Then, the mixture solutions were studied with a XenoScreen YES/YAS assay to determine the variations in the initial hormonal response of each pharmaceutical compound due to the presence of a bisphenol analog. The results obtained were modeled with the concentration addition (CA) and independent action (IA) approaches, the trueness of which was studied with model deviation ratios (MDR). The estrogenic agonistic activity of the drugs studied was most strongly affected by the presence of BPA in solution (twenty-one cases of synergy observed for CA models versus twelve cases of antagonism in the case of IA predictions). BPS shows a strong agonistic estrogenic impact on most of the drugs studied at medium and high concentration levels; androgenic agonistic activity was also impaired with elevated concentrations of BPS.

Genotoxicity of selected pharmaceuticals, their binary mixtures, and varying environmental conditions – study with human adenocarcinoma cancer HT29 cell line

Pharmaceutical residues are present in the environment in mixtures and their adverse effects may also result from interactions that occur between compounds. Studies presented in this work focus on genotoxicity of pharmaceuticals from different therapeutic groups in mixtures and in individual solutions impacted with different environmental conditions assessed using comet assay (alkaline approach). Binary mixtures of pharmaceuticals (in different concentration ratios) and in individual solutions impacted with pH change (range from 5.5 to 8.5) or addition of inorganic ions, were incubated with HT29 cells and after 24 h time period cells were tested for the presence of DNA damage. To estimate whether mixtures act more (synergistic) or less (antagonistic) efficiently Concentrations Addition (CA) and Independent Action (IA) approaches were applied followed by a calculation of the Model Deviation Ratio (MDR) to determine deviation from the predicted values. Addition of inorganic ions mainly reduced their genotoxicity. Diclofenac s. was the most susceptible to potassium, fluoride, and bromide ions. Change of the pH of pharmaceutical solutions had significant impact on genotoxicity of diclofenac s. and fluoxetine h. Among mixtures, more commonly observed interactions were synergistic ones, exactly twenty-five cases (ten pairs containing chloramphenicol or oxytetracycline h.) and ten cases of antagonism (four for pairs containing chloramphenicol or fluoxetine h.). The results obtained indicate that interactions between tested compounds occur frequently and can lead to DNA damage. This topic especially concerning in vitro tests using cells is still rare, however, it should not be neglected.

Influence of Storage Time and Temperature on the Toxicity, Endocrine Potential, and Migration of Epoxy Resin Precursors in Extracts of Food Packaging Materials

B. Kudłak , N. Jatkowska , P. Kubica , G. Yotova, T. Stefan – MOLECULES – 2019
The aim of the present study was to establish a standard methodology for the extraction of epoxy resin precursors from several types of food packages (cans, multi-layered composite material, and cups) with selected simulation media (distilled water, 5% ethanol, 3% dimethyl sulfoxide, 5% acetic acid, artificial saliva) at different extraction times and temperatures (factors). Biological analyses were conducted to determine the acute toxicity levels of the extracts (with Vibrio fischeri bacteria) and their endocrine potential (with Saccharomyces cerevisiae yeasts). In parallel, liquid chromatography-tandem mass spectrometry was performed to determine levels of bisphenol A diglycidyl ether (BADGE), bisphenol F diglycidyl ether (mixture of isomers, BFDGE), ring novolac glycidyl ether (3-ring NOGE), and their derivatives. The variation induced by the different experimental factors was statistically evaluated with analysis of variance simultaneous component analysis (ASCA). Our findings demonstrate the value of using a holistic approach to best partition the effects contributing to the end points of these assessments, and offer further guidance for adoptingsuch a methodology, thus being a broadly useful reference for understanding the phenomena related to the impacts of food packaging materials on quality for long- and short-term storage, while offering a general method for analysis.

Stabilities of bisphenol A diglycidyl ether, bisphenol F diglycidyl ether, and their derivatives under controlled conditions analyzed using liquid chromatography coupled with tandem mass spectrometry

Bisphenol A diglycidyl ether (BADGE), bisphenol F diglycydyl ether (BFDGE), and their related compounds are widely used as precursors in production of epoxy resins. The high reactivity of these compounds makes the development of analytical methodologies that ensure appropriate metrological accuracy crucial. Consequently, we aimed to determine whether and to what extent the composition of the solution and storage conditions affect the stability of selected BADGE and BFDGE derivatives. The stabilities of these compounds were studied using liquid chromatography–tandem mass spectrometry with electrospray ionization (HPLC-ESI–MS/MS). The chromatographic method elaborated here has allowed for separation of the analytes in time shorter than 6 min, for both methanol and acetonitrile-based mobile phases. The obtained calibration curves for all analytes were linear in the range tested. The values of limit of detection (LODs) were in the range of 0.91–2.7 ng/mL, while values of limit of quantitation (LOQs) were in the range of 2.7–5.7 ng/mL. The chosen experimental conditions were compared in terms of the content of organic solvent in solution, storage temperature, and time. Our results show that the content of BADGE, BADGE·HCl, BFDGE, three-ring NOGE decreased with increasing water content (> 40% v/v). For BADGE and three-ring NOGE, significant changes in concentration were noted as early as 24 h after the test solutions had been prepared. In addition, a reduction in the storage temperature (4 to − 20 °C) reduced the rate of transformation of the monitored analytes. Our study will increase quality control in future research and may increase the reliability of the obtained results.

Assessing Acute Toxicity and Endocrine Disruption Potential of Selected Packages Internal Layers Extracts

N. Jatkowska , B. Kudłak , G. Yotova, S. Tsakovski, J. Namieśnik – 2018
The focus of the present study is migration of toxic substances from food contact materials and its actual influence on the health of the final consumer. Two food packagings (metal cans, TetraPack®) and five simulants medias (water, ethanol, acetic acid, DMSO, artificial saliva) were selected for simulation studies. For assessment of acute toxicity and endocrine disruption potential of extract samples, two biotests (Microtox® and XenoScreen YES/YAS) were performed. Multi-factor analysis of variation (MANOVA) was used to evaluate the effects of the three main factors - solvent, temperature, contact time and their interactions on the respected dependent variable (acute toxicity or estrogen disruption potential).

Assessing ecotoxicity and the endocrine potential of selected phthalates, BADGE and BFDGE derivatives in relation to environmentally detectable levels

There is no doubt that the subject area of plasticmaterials (e.g., production of epoxy resins or polyesters) is inherently connected to issues concerning bisphenol A (BPA) and its analogues. Unfortunately, much less attention has been given to other compounds, which are also used for the production of these materials. Bisphenol A diglycidyl ether (BADGE) is a synthetic industrial compound obtained by a condensation reaction between epichlorohydrin (ECH) and BPA. Similarly, novolac glycidyl ether (BFDGE) is produced in thereactionbetween novolac and epichlorohydrin. Nevertheless, there is a lack of information on the combined effects of BADGE derivatives at environmentally relevant levels. In the current study, toxicity levels inMicrotox® and XenoScreen YES/YAS assayswere determined for several analogues alone, then the biological effects of compound pairs mixed in 33, 66 and 100% of each compounds' EC50 ratios were evaluated. The Microtox® test has been chosen as a relevant tool, and the results were referred to the Xenoscreen YES/YAS assay,which has been chosen for the fast determination of the endocrine potential of the compounds tested. The results obtained constitutes the basis for model studies, with Concentration Addition (CA) and Independent Action (IA), followed by Model Deviation Ratio (MDR) interpretation, to evaluate the possible interactions occurring between analytes when present inmixtures. The results indicate that the hydrochloric derivatives of BADGE and BFDGE are of the greatest toxicological and endocrine threat. Thus, their presence inmixtures under certain environmental conditions (including presence in the tissues of living organisms) should be strictly monitored and reported, especially in acidic environments. Strong evidence on the synergic behaviors of these analytes, which expressed high toxicity (EC50 2.69–117.49 μg/mL), is demonstrated with Model Deviation Ratio (MDR).