Acute and sub-chronic toxicity of four cytostatic drugs in zebrafish

The acute and sub-chronic effects of four cytostatic drugs—5-fluorouracil (5-FU), cisplatin (CisPt), etoposide (ET) and imatinib mesylate (IM)—on zebrafish (Danio rerio) were investigated. Acute tests were carried out in a static system in accordance with the OECD guideline 203 for adult fish and the draft guideline for fish embryos (FET test) in order to find the LC50 values of the four cytostatic drugs. Early-life stage toxicity test on zebrafish was conducted according the OECD guideline 210 using the cytostatic drugs 5-FU and IM in a semistatic system with the objective of investigating the sub-chronic effects of the cytostatic drugs on fish. In adult fish, the cytostatic drugs 5-FU and ET did not pass the limit test, thus, are considered non-toxic. In case of cisplatin, LC50 was calculated at 64.5 mg L^?1, whereas in case of IM, LC50 was at 70.8 mg L^?1. In the FET test, LC50 of 5-FU at 72-h post fertilization (hpf) was 2441.6 mg L^?1. In case of CisPt, LC50 was 349.9 mg L^?1 at 48 hpf and it progressively decreased to 81.3 mg L^?1 at 120 hpf. In addition, CisPt caused a significant delay in the hatch of larvae. In case of ET, LC50 values were not calculable as they were higher than 300 mg L^?1 at which concentration the substance crystallized in the solution. LC50 values of IM were 48 hpf; 158.3 mg L^?1 , 72 hpf; 141.6 mg L^?1, 96 hpf; 118.0 mg L^?1, and 120 hpf; 65.9 mg L^?1. In the Early-life Stage Test with 5-FU, embryonic deformities were not detected during the tests. Regarding mortalities, the 10 mg L^?1 concentration can be considered as LOEC, as statistically significant difference in mortalities was detected in this group alone. Concerning dry body weight and standard length, 1 mg L^?1 is the LOEC. In case of IM, the highest tested concentration (10 mg L^?1) can be considered LOEC for mortalities, however, the treatment did not have an effect on the other investigated parameters (dry and wet weight, standard length). All four cytostatic drugs were characterized by low toxicity in zebrafish in acute and sub-chronic tests.     

Genotoxic and teratogenic potential of marine sediment extracts investigated with comet assay and zebrafish test

Organic extracts of marine sediments from the North Sea and the Baltic Sea were investigated with two toxicity assays. The comet assay based on the fish cell line Epithelioma papulosum cyprini (EPC) was applied to determine the genotoxic potential; zebrafish embryos (Danio rerio) were used to quantify the teratogenic potential of the samples. EC(50) values were calculated from dose-response curves for both test systems. Highest teratogenic and genotoxic effects normalised to total organic carbon (TOC) content were detected in sediment samples of different origins. Polychlorinated biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs) are not likely to be the causes of the observed effects, as demonstrated by a two-step fractionation procedure of selected extracts. The toxic potential was more pronounced in fractions having polarity higher than those possessed by PAHs and PCBs. The suitability of the two in vitro test systems for assessing genotoxic and teratogenic effects of marine sediment extracts could be demonstrated.     

Impact of common cytostatic drugs on pollen fertility in higher plants

Cytostatic drugs are among the most toxic chemicals which are produced. Many of them cause damage of the genetic material which may affect the fertility of higher organisms. To study the impact of the widely used anticancer drugs [cisplatin (CisPt), etoposide (Et), and 5-fluorouracil (5-FU)] on the reproduction of higher plants, pollen abortion experiments were conducted with species which belong to major plant families, namely with Tradescantia paludosa (Commelinaceae), Arabidopsis thaliana (Brassicaceae), Chelidonium majus (Papaveraceae), and Alisma plantago-aquatica (Alismataceae). All compounds increased the frequencies of abortive grains. The lowest effective doses were in general in a narrow range (i.e., 1 and 10 mg/kg of dry soil). The effects of the individual drugs were similar in T. paludosa, A. plantago-aquatica, and Ch. majus, while A. thaliana was consistently less sensitive. The highest abortion rate was obtained in most experiments with CisPt, followed by 5-FU and Et. Comparisons of the doses which caused effects in the present experiments in the different species with the predicted environment concentrations and with the levels of the cytostatics which were detected in hospital wastewaters show that the realistic environmental concentrations of the drugs are 4–6 orders of magnitude lower. Therefore, it is unlikely that these drugs affect the fertility of higher plants in aquatic and terrestrial ecosystems.     

Influence of selected anti-cancer drugs on the induction of DNA double-strand breaks and changes in gene expression in human hepatoma HepG2 cells

In chemotherapy, various anti-cancer drugs with different mechanisms of action are used and may represent different risk of undesirable delayed side effects in treated patients as well as in occupationally exposed populations. The aim of the present study was to evaluate genotoxic potential of four widely used anti-cancer drugs with different mechanisms of action: 5-fluorouracil (5-FU), cisplatin (CDDP) and etoposide (ET) that cause cell death by targeting DNA function and imatinib mesylate (IM) that inhibits targeted protein kinases in cancer cells in an experimental model with human hepatoma HepG2 cells. After 24 h of exposure all four anti-cancer drugs at non-cytotoxic concentrations induced significant increase in formation of DNA double strand breaks (DSBs), with IM being the least effective. The analysis of the changes in the expression of genes involved in the response to DNA damage (CDKN1A, GADD45A, MDM2), apoptosis (BAX, BCL2) and oncogenesis (MYC, JUN) showed that 5-FU, CDDP and ET upregulated the genes involved in DNA damage response, while the anti-apoptotic gene BCL2 and oncogene MYC were downregulated. On the contrary, IM did not change the mRNA level of the studied genes, showing different mechanism of action that probably does not involve direct interaction with DNA processing. Genotoxic effects of the tested anti-cancer drugs were observed at their therapeutic concentrations that may consequently lead to increased risk for development of delayed adverse effects in patients. In addition, considering the genotoxic mechanism of action of 5-FU, CDDP and ET an increased risk can also not be excluded in occupationally exposed populations. The results also indicate that exposure to 5-FU, CDDP and ET represent a higher risk for delayed effects such as cancer, reproductive effects and heritable disease than exposure to IM.     

An evaluation of the cytochrome P450 inhibition potential of selected pesticides in human hepatic microsomes

The goal of this work was to study the ability of 18 pesticides to inhibit selective model activities for all major xenobiotic-metabolizing enzymes, namely CYP1A1/2, 2A6, 2B6, 2C8, 2C9, 2C19, 2D6, 2E1 and 3A4. Generally organophosphorus insecticides were the most potent and extensive inhibitors, especially towards CYP1A1/2 (IC₅₀ values of chlorpyrifos, fenitrothion and profenofos ～3 μ M), CYP2B6 (IC₅₀ values of chlorpyrifos and fenitrothion 2.5 μ M), CYP2C8 (fenitrothion 4.3 μ M), CYP2C9 (fenitrothion and malathion 4.8 and 2.5 μ M, respectively), CYP2D6 (chlorpyrifos and phenthoate ～ 3 μ M) and CYP3A4 (chlorpyrifos, fenitrothion and phenthoate 3–4 μ M). Otherwise there were quite considerable differences in potency and extent of inhibition between different organophosphates. Pyrethroids were in general very weak or inactive. Deltamethrin and fenvalerate were potent inhibitors of CYP2D6 (IC₅₀ values of ～ 3 μ M) while lambda-cyhalothrin potently inhibited both CYP2D6 and CYP3A4-mediated activities (IC₅₀'s about 3–4 μ M). Some pesticides caused relatively potent inhibitions sporadically (carbendazim, CYP2D6, IC₅₀ = 12 μ M; atrazine, CYP3A4, IC₅₀ = 2.8 μ M; glyphosate, CYP2C9, IC₅₀ = 3.7 μ M; hexaflumuron, IC₅₀ = 6.0 μ M). With the exceptions of alpha-cypermethrin, cypermethrin, isoproturon, carbaryl and abamectin, most pesticides inhibited relatively potently at least one CYP-selective activity, which may have relevance for potential interactions in occupational exposures and for further studies on the CYP-associated metabolism of respective pesticides.     

Assessing copper thresholds for phytotoxicity and potential dietary toxicity in selected vegetable crops

Copper pollution in soils is widespread, and its accumulation in crop products could pose a risk on human health. In this paper, bioavailability of added copper (Cu) and critical Cu concentrations in a vegetable garden soil was evaluated for Chinese cabbage (Brassica chinensis L.), pakchoi (Brassica chinensis L.), and celery (Apiumg graveolens L. var. dulce DC) based on human dietary toxicity. The availability of added Cu in the soil decreased with incubation time, and had minimal change after 10-12 weeks. After incubated for 12 weeks, about 60% of added Cu was not extractable by DTPA. The same crops were also grown in sand culture to determine their responses to solution Cu. Shoot growth was significantly inhibited at Cu concentrations above 10 mg kg(-1) in the solution or above 150 mg kg(-1) (DTPA-Cu) in the soil. The sensitivity of the crops to Cu toxicity differed among the three vegetable crops. Copper concentration in shoots and edible parts varied with Cu supply levels and type of the vegetables. Negative correlations (r=-0.90-0.99**) were noted between Cu concentration in shoots and fresh matter yields, but Cu concentrations in the edible parts were positively correlated with available and total Cu in the soil (r=0.91-0.99**). The critical tissue Cu concentrations at 10% shoot DM reduction were 19.4, 5.5, 30.9 mg kg(-1) for Chinese cabbage, pakchoi, and celery, respectively. Based on the threshold of human dietary toxicity for Cu (10 mg kg(-1)), the critical concentrations of total and available Cu in the soil were 430 and 269 mg kg(-1) for pakchoi, 608 and 313 mg kg(-1) for celery, and 835 and 339 mg kg(-1) for Chinese cabbage, respectively.     

Toxicity of the mixture of selected antineoplastic drugs against aquatic primary producers

The residues of antineoplastic drugs are considered as new and emerging pollutants in aquatic environments. Recent experiments showed relatively high toxicity of 5-fluorouracil (5-FU), imatinib mesylate (IM), etoposide (ET) and cisplatin (CP) that are currently among most widely used antineoplastic drugs, against phytoplankton species. In this study, we investigated the toxic potential of the mixture of 5-FU?+?IM?+?ET against green alga Pseudokirchneriella subcapitata and cyanobacterium Synechococcus leopoliensis, and the stability and sorption of these drugs to algal cells. Toxic potential of the mixture was predicted by the concepts of ‘concentration addition’ and ‘independent action’ and compared to the experimentally determined toxicity. In both test species, the measured toxicity of the mixture was at effects concentrations EC₁₀–EC₅₀ higher than the predicted, whereas at higher effect concentration (EC₉₀), it was lower. In general, P. subcapitata was more sensitive than S. leopoliensis. The stability studies of the tested drugs during the experiment showed that 5-FU, IM and CP are relatively stable, whereas in the cultures exposed to ET, two transformation products with the same mass as ET but different retention time were detected. The measurements of the cell-linked concentrations of the tested compounds after 72 h exposure indicated that except for CP (1.9 % of the initial concentration), these drugs are not adsorbed or absorbed by algal cells. The results of this study showed that in alga and cyanobacteria exposure to the mixture of 5-FU?+?ET?+?IM, in particular at low effect concentration range, caused additive or synergistic effect on growth inhibition, and they suggest that single compound toxicity data are not sufficient for the proper toxicity prediction for aquatic primary producers.     

Adsorption,desorption, potential and selective distribution op heavy metals in selected soils of japan

Abstract

Adsorption, desorption, potential and selective distribution of Cu, Zn, Cd, Pb and Ni were investigated in three typical soils of Japan under flooded condition.

The results indicate that the sorption of all heavy metals was linear upto the maximum concentration (500 μg/g soil) employed in the present studies in all the soils. The magnitude of sorption in general was in the order of Pb > Cu > Zn > Cd > Ni. The adsorption coefficients showed wide variations among different soils as well as metal ions. The hysteresis of sorption and desorption by KNO₃ was well pronounced for both the metal ions and the soils. The desorption rate was greater than the fixation rate indicating the predominance of the chemosorption over physical processes. The major portion of sorbed metals were retained in the unextractable form, which over all accounted for more than 50% of the sorbed metals.     

The antiproliferative and apoptotic potential of Clinacanthus nutans against human breast cancer cells through targeted apoptosis pathway

Environmental Science and Pollution Research - Clinacanthus nutans dichloromethane fraction (CN-Dcm) extract has previously been proven to suppress breast cancer (MCF7) cell proliferation. Despite...     

The relation between levels of selected PCB congeners in human serum and follicular fluid

Individual congener and total PCB concentrations were determined in serum and follicular fluid obtained from women undergoing assisted reproductive technologies (in-vitro fertilization and embryo replacement). Although the mean individual PCB levels revealed varying degrees of contamination, the results fall in the same range as that observed by other investigators. Except for PCB 118, correlations between levels in serum and follicular fluid were strong, and statistically significant at p < 0.05. Moreover PCB 153, a major and very stable PCB congener has been shown to correlate to the total amount of PCBs (r = 0.994, and r = 0.987, for serum and follicular fluid, respectively). The same accumulation patterns of PCBs for serum and follicular fluid have been observed.     

Genotoxic and cytotoxic effects of pethoxamid herbicide on Allium cepa cells and its molecular docking studies to unravel genotoxicity mechanism

Environmental Science and Pollution Research - Pethoxamid is chloroacetamide herbicide. Pethoxamid is commonly used to kill different weeds in various crops. Pethoxamid can leach in the water and...     

Genotoxic effect of ciprofloxacin during photolytic decomposition monitored by the in vitro micronucleus test (MNvit) in HepG2 cells

Purpose

Ciprofloxacin (CIP), a broad-spectrum, second-generation fluoroquinolone, has frequently been found in hospital wastewaters and effluents of sewage treatment plants. CIP is scarcely biodegradable, has toxic effects on microorganisms and is photosensitive. The aim of this study was to assess the genotoxic potential of CIP in human HepG2 liver cells during photolysis.

Methods

Photolysis of CIP was performed in aqueous solution by irradiation with an Hg lamp, and transformation products were monitored by HPLC-MS/MS and by the determination of dissolved organic carbon (DOC). The cytotoxicity and genotoxicity of CIP and of the irradiated samples were determined after 24?h of exposure using the WST-1 assay and the in vitro micronucleus (MN) test in HepG2 cells.

Results

The concentration of CIP decreased during photolysis, whereas the content of DOC remained unchanged. CIP and its transformation products were not cytotoxic towards HepG2 cells. A concentration-dependent increase of MN frequencies was observed for the parent compound CIP (lowest observed effect level, 1.2???mol?L^?1). Furthermore, CIP and the irradiated samples were found to be genotoxic with a significant increase relative to the parent compound after 32?min (P?P?Conclusions Photolytic decomposition of aqueous CIP leads to genotoxic transformation products. This proves that irradiated samples of CIP are able to exert heritable genotoxic effects on human liver cells in vitro. Therefore, photolysis as a technique for wastewater treatment needs to be evaluated in detail in further studies, not only for CIP but in general.     

Genotoxic potentials and related mechanisms of bisphenol A and other bisphenol compounds: A comparison study employing chicken DT40 cells

Bisphenol A (BPA) has been found in plastic food containers, paper currencies and toys. BPA has been reported for various adverse health concerns including reproduction, development and carcinogenesis. These potential health implications have led to increasing use of alternative bisphenols such as bisphenol F and bisphenol S among many. However, little is known about the toxicity of alternative bisphenols and most of the toxicological information is limited to endocrine disrupting potentials. In this study, we evaluated cytotoxicity and the genotoxic potentials of several bisphenol compounds, and identified the mechanism of genotoxicity using a panel of mutant chicken DT40 cell lines deficient in DNA repair pathways. Several bisphenols including bisphenol AP, bisphenol M, or bisphenol P exerted genotoxic potentials that are greater than that of BPA. Generally RAD54^−/− mutant cells were the most sensitive to all bisphenols except for bisphenol F, suggesting the induction of DNA double-strand breaks that could be rescued by homologous recombination. Genotoxic potential of bisphenols was confirmed by chromosomal aberration assay and γ-H2AX foci forming assay between wild-type and RAD54^−/− mutant. Among the tested bisphenols, BPP at 12.5 μM showed the greatest genotoxic potency, inducing chromosomal aberration and γ-H2AX foci in RAD54^−/− mutant by 2.6 and 4.8 folds greater than those in wild-type, respectively. Our results clearly show several alternative bisphenols can cause genotoxicity that could be rescued by homologous recombination pathway, and some bisphenols induced even greater genotoxic potentials than that of BPA.     

Analyses of combined effects of cytostatic drugs on micronucleus formation in the <Emphasis Type="Italic">Tradescantia</Emphasis>

Recent experiments showed that 5-fluorouracil (5FU), cisplatin (CDDP), etoposide (ET), and imatinib mesylate (IM), which are currently among the most widely used anticancer drugs, cause damage of the genetic material in higher plants. The aim of the present study was to determine whether mixtures of these drugs cause synergistic or antagonistic effects which may have an impact on their environmental safety. Therefore, the effects of binary mixtures of these anticancer drugs on the induction of micronuclei (MN) which reflect structural and numerical chromosomal aberrations were assessed in Tradescantia tetrads. Synergistic/antagonistic effects were determined by comparison with single exposures that would be equally effective in a reference model of independent action. This comparison was performed at two distinct effect sizes. We found clear evidence for synergisms in combination experiments with IM and antagonism in a high-dose experiment with ET and 5FU. Our findings indicate that IM increases the genotoxic effects of other anticancer drugs. The maximal effects which we found were in the range between 19 and 38 % in the excess of effect sizes predicted under independent action. These effects may have an impact on the overall genotoxic activities of untreated hospital waste waters but not on the environment in general as the predicted environmental concentrations of the studied drugs are several orders of magnitude lower as the levels which are required to cause induction of MN in higher plants.     

Biofilm controlled sorption of selected acidic drugs on river sediments characterized by different organic carbon content

The sorption process of selected non-steroidal anti-inflammatory drugs (ibuprofen, naproxen, ketoprofen, diclofenac) on biofilm covered river sediments were investigated in laboratory. In the course of the experiments, the effect of pH of aqueous phase, the effect of TOC (total organic carbon) content of biofilm on the sorption processes were studied. The determination of concentration of drugs was performed by gas chromatography mass spectrometry (GC-MS) both in liquid and solid phases. The pseudo-first-order rate constant of the sorption was found to be 83 min(-1). The effect of pH on the sorption of diclofenac was significantly lower than the obtained values in case of the other three drugs. The calculated K(d) (sorption coefficient) values increased in the sequence of ibuprofen, naproxen, ketoprofen and diclofenac and varied between 0.1-0.4; 0.2-0.7; 0.2-1.2; 0.2-1.4 kg L(-1) respectively, depending on the characteristics of the sediments. The value of K(d)×f(oc) showed a straight line as function of f(oc) (fraction of organic carbon) therefore, instead of the widely distributed normalization process (K(d)/f(oc)), an empirical equation (K(d)=A/f(oc)+B) was suggested for estimation of the K(d) values in case of different TOC content sediments.     

Cypermethrin has the potential to induce hepatic oxidative stress, DNA damage and apoptosis in adult zebrafish (Danio rerio)

Cypermethrin (CYP), a widely used Type II pyrethroid pesticide, is one of the most common contaminants in the freshwater aquatic system. We studied the effects of CYP exposure on the induction of hepatic oxidative stress, DNA damage and the alteration of gene expression related to apoptosis in adult zebrafish. Hepatic mRNA levels for the genes encoding antioxidant proteins, such as Cu/Zn-Sod, Mn-Sod, Cat, and Gpx, were significantly upregulated when zebrafish were exposed to various concentrations of CYP for 4 or 8 days. In addition, the main genes related to fatty acid β-oxidation and the mitochondrial genes related to respiration and ATP synthesis were also significantly upregulated after exposure to high concentrations (1 and 3 μg L⁻¹) of CYP for 4 or 8 days. Moreover, in a comet assay of zebrafish hepatocytes, tail DNA, tail length, tail moment and Olive tail moment increased in a concentration-dependent manner. The significant induction (p < 0.01) of all four parameters observed with CYP concentrations of 0.3 μg L⁻¹ or higher suggests that heavy DNA damage was induced even at low levels. Furthermore, several apoptosis- related genes, such as p53, Apaf1 and Cas3, were significantly upregulated after CYP exposure, and Bcl2/Bax expression ratio decreased, especially in groups treated with 1 and 3 μg L⁻¹ CYP for 8 days. Taken together, our results suggested that CYP has the potential to induce hepatic oxidative stress, DNA damage and apoptosis in zebrafish. This information will be helpful in fully understanding the mechanism of aquatic toxicology induced by CYP in fish.     

Changes in extreme events and the potential impacts on human health

Extreme weather and climate-related events affect human health by causing death, injury, and illness, as well as having large socioeconomic impacts. Climate change has caused changes in extreme event frequency, intensity, and geographic distribution, and will continue to be a driver for change in the future. Some of these events include heat waves, droughts, wildfires, dust storms, flooding rains, coastal flooding, storm surges, and hurricanes. The pathways connecting extreme events to health outcomes and economic losses can be diverse and complex. The difficulty in predicting these relationships comes from the local societal and environmental factors that affect disease burden. More information is needed about the impacts of climate change on public health and economies to effectively plan for and adapt to climate change. This paper describes some of the ways extreme events are changing and provides examples of the potential impacts on human health and infrastructure. It also identifies key research gaps to be addressed to improve the resilience of public health to extreme events in the future.

Implications: Extreme weather and climate events affect human health by causing death, injury, and illness, as well as having large socioeconomic impacts. Climate change has caused changes in extreme event frequency, intensity, and geographic distribution, and will continue to be a driver for change in the future. Some of these events include heat waves, droughts, wildfires, flooding rains, coastal flooding, surges, and hurricanes. The pathways connecting extreme events to health outcomes and economic losses can be diverse and complex. The difficulty in predicting these relationships comes from the local societal and environmental factors that affect disease burden.     

Cellular uptake and mutagenic potential of metal oxide nanoparticles in bacterial cells

Extensive production and consumption of nanomaterials such as ZnO and TiO₂ has increased their release and disposal into the environment. The accumulation of nanoparticles (NPs) in ecosystem is likely to pose threat to non-specific targets such as bacteria. The present study explored the effect of ZnO and TiO₂ NPs in a model bacterium, Salmonella typhimurium. The uptake of ZnO and TiO₂ bare NPs in nano range without agglomeration was observed in S. typhimurium. TEM analysis demonstrated the internalization and uniform distribution of NPs inside the cells. Flow cytometry data also demonstrates that both ZnO and TiO₂ NPs were significantly internalized in the S. typhimurium cells in a concentration dependent manner. A significant increase in uptake was observed in the S. typhimurium treated even with 8 and 80 ng mL⁻¹ of ZnO and TiO₂ NPs with S9 after 60 min, possibly the formation of micelles or protein coat facilitated entry of NPs. These NPs exhibited weak mutagenic potential in S. typhimurium strains TA98, TA1537 and Escherichia coli (WP2uvrA) of Ames test underscoring the possible carcinogenic potential similar to certain mutagenic chemicals. Our study reiterates the need for re-evaluating environmental toxicity of ZnO and TiO₂ NPs presumably considered safe in environment.     

Global potential of phosphorus recovery from human urine and feces

This study geospatially quantifies the mass of an essential fertilizer element, phosphorus, available from human urine and feces, globally, regionally, and by specific country. The analysis is performed over two population scenarios (2009 and 2050). This important material flow is related to the presence of improved sanitation facilities and also considers the global trend of urbanization. Results show that in 2009 the phosphorus available from urine is approximately 1.68 million metric tons (with similar mass available from feces). If collected, the phosphorus available from urine and feces could account for 22% of the total global phosphorus demand. In 2050 the available phosphorus from urine that is associated with population increases only will increase to 2.16 million metric tons (with similar mass available from feces). The available phosphorus from urine and feces produced in urban settings is currently approximately 0.88 million metric tons and will increase with population growth to over 1.5 million metric tons by 2050. Results point to the large potential source of human-derived phosphorus in developing regions like Africa and Asia that have a large population currently unserved by improved sanitation facilities. These regions have great potential to implement urine diversion and reuse and composting or recovery of biosolids, because innovative technologies can be integrated with improvements in sanitation coverage. In contrast, other regions with extensive sanitation coverage like Europe and North America need to determine how to retrofit existing sanitation technology combined that is combined with human behavioral changes to recover phosphorus and other valuable nutrients.     

Correction to: Genotoxic and cytotoxic effects of pethoxamid herbicide on Allium cepa cells and its molecular docking studies to unravel genotoxicity mechanism

Environmental Science and Pollution Research -