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.

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.

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.

Determination of Toxicological Parameters of Selected Bioactive Organic Chemicals Using the Ostracodtoxkit FTM

Assessment of the impact of pharmaceutical residues on living organisms is very complex subject. Apart from taking into account the toxicity of individual compounds also their presence in mixtures should be taken into account. In this work, attempts were made to assess the ecotoxicity of biologically active substances (with 50 % effective concentration (EC50) values growing from fluoxetine (EC50 = 4.431 nM) >> gemfibrozil ≈ 17α-ethinylestradiol ≈ ketorolac > indomethacin > theophylline ≈ progesterone > naproxen ≈ trypsin > 2-(2,4,5-trichlorophenoxy)propionic acid > chloramphenicol > acetylsalicylic acid > ibuprofen > ketoprofen > 19-norethindrone to bezafibrate as the least toxic drug among studied ones) to the ISO standardized Ostracodtoxkit FTM bioassay. The Ostracodtoxkit FTM was proven to be very sensitive tool with respect to responding to presence of pharmaceuticals.

Impact of inorganic ions and pH variations on toxicity and endocrine potential of selected environmentally relevant pharmaceuticals

M. Wieczerzak , B. Kudłak , G. Yotova, S. Tsakovski, V. Simeonov, J. Namieśnik ENVIRONMENTAL POLLUTION – 2018
Assessment of the impact of pharmaceutical residues on living organisms is a very complex subject. Apart from taking into account the toxicity of individual compounds, environmental factors should also be taken into account. In this paper, attempts were made to assess the impact of coexisting inorganic ions and changes in pH on the toxicity of ten selected pharmaceuticals. Two bioassays were used to measure the estrogenic and androgenic effects (XenoScreen YES/YAS - Saccharomyces cerevisiae) and acute toxicity (Microtox® - Vibrio fischeri). The Microtox® test gave the most definitive outputs concerning the determination of interaction type between drugs and chemical species. Synergism was proven for almost all drugs and chemical species, and only two cases of antagonism were found. Significant drug/pH interactions were rare. Regarding the XenoScreen YES/YAS bioassay, when estrogenic and androgenic agonistic effects (YES+ and YAS+, respectively) were studied, many cases of well-expressed synergism for all inorganic ions with limited number of drugs (diazepam, fluoxetine, estrone, chloramphenicol for the YES+ test and diazepam, progesterone, androstenedione, and estrone for the YAS+ test) were found. Antagonism was also proven for the YES+ test, especially for diclofenac and androstenedione interacting with cations. On the other hand, the YES- and YAS- tests (estrogenic and androgenic, respectively, antagonistic effects) did not indicate cases of synergetic interaction except for the couples Br-/diazepam and NH4+/ketoprofen. Antagonistic drug/ion interactions were detected only with diclofenac and fluoxetine. It is interesting that well-expressed (antagonism or synergism) drug/pH interactions were rare. Both tests were found utilizable in performing studies on impact of ions/pH fluctuations on drugs mixtures' toxicity confirming in most cases synergic impact of parameters studied on toxicity. The approach proposed in the paper seems to be proven as a reliable tool in assessing impact of abiotic factors on toxicity and endocrine potential of complex mixtures of pharmaceuticals' mixtures.

Impact of selected drugs and their binary mixtures on the germination of Sorghum bicolor (sorgo) seeds

The aim of this study was to assess the toxicological potential of binary drug mixtures and individual drugs under different pH conditions with different inorganic ion additions on the germination of Sorghum bicolor (sorgo) seeds. To assess whether the given drug mixtures were more phytotoxic than the individual compounds, concentration addition (CA) and independent action (IA) approaches were applied to estimate the predicted phytotoxicity values, followed by calculations of the MDR (model deviation factor) to determine the deviation from the predicted values. Binary mixtures of chloramphenicol with ketoprofen, diclofenac sodium, and oxytetracycline hydrochloride proved to be synergistic. The pH changes had the largest impact on the phytotoxicity of the diclofenac sodium and ketoprofen solutions, elevating their toxicity toward S. bicolor, and the co-presence of inorganic ions was shown to have an impact on ketoprofen, chloramphenicol, and oxytetracycline hydrochloride. Most of the interactions between sorgo plants and pharmaceuticals with added ions were antagonistic in nature, particularly those calculated using the IA model, with a few cases (one case for ketoprofen and chloramphenicol, two cases for oxytetracycline hydrochloride, and four cases for diclofenac sodium) of overestimation.

Cyanogenic Compounds and Estrogen Disruptors

Balanced diet consists largely of plants containing cyanogenic compounds in the form of the more common cyanogenic glycosides and sometimes lipids. Maize, wheat, rye, apples, barley, oats, sugar cane and yet many other plants consumed by humans contain cyanogenic compounds. However the risk of poisoning is negligible as it is very easy to remove the toxic HCN by grinding and drying in air or soaking in water and an additional thermal treatment contributes to the denaturation of proteins (enzymes) that initiate process of cyanogenesis. Furthermore, man is provided with a quite effective cyanide detoxifying mechanisms, functioning effectively with adequate protein diet. In 1990, scientists suggested that the role of cyanogenic compounds can only come down to defense against fungi because their harmfulness is negligible. Since little evidences exist on possibility of cyanogenic compounds as direct EDC, studies are needed to confirm possibility of their impact on hormonal action of higher animals due to binding with hormones, their degradation or metabolic shifts after HCN release into circulation system.

Toksyczność - parametr sumaryczny do oceny obciążenia danego elementu środowiska przez ksenobityki

Wyemitowane zanieczyszczenia pochodzenia antropogenicznego przedostają się do poszczególnych elementów środowiska, gdzie mogą negatywnie oddziaływać na organizmy zwierzęce i roślinne. Systematyczny monitoring stopnia zanieczyszczenia środowiska powiązany z szybkim systemem reagowania wydaje się jednym z lepszych rozwiązań kwestii kontroli jakości środowiska. Jednak klasyczne metody instrumentalnej analizy próbek środowiskowych nie zawsze są środowiskowo przyjazne za to często są kosztowne oraz charakteryzują się potrzebą użycia toksycznych rozpuszczalników, dodatkowo nie zawsze istnieje konieczność i możliwość oznaczenia wszystkich składników badanej próbki. Z tej przyczyny wydaje się być zasadnym wykorzystanie testów toksykologicznych, jako parametru sumarycznego do określenia stanu danego elementu środowiska. Nowoczesne techniki bioanalityczne umożliwiają uzyskanie ilościowych i jakościowych informacji o badanej próbce środowiskowej, nie wyklucza to jednak zastosowania technik instrumentalnych, dlatego najwydajniejszym rozwiązaniem byłoby połączenie danych uzyskanych z obu źródeł, celem uzyskania pełniejszej informacji o stanie środowiska.