The natural anthraquinone dye emodin: Eco/genotoxicological characterization for aquatic organisms.

Emodin is an anthraquinone secondary metabolite produced by several species of plants and fungi. Emodin is known for its pharmacological versatility, and, in the textile industry, for its good dyeing properties. However, its use in the textile industry can result in the formation and disposal of large volumes of wastewater. Emodin mutagenicity has been shown in bacteria and in human cells, but little is known about its possible toxic, genotoxic, or mutagenic effects in aquatic organisms. We have evaluated the eco/genotoxicity of emodin to aquatic organisms. Emodin was toxic to Daphnia similis (EC50 = 130 µg L-1) and zebrafish embryos (LC50 = 25 µg L-1). No toxicity was observed for Raphidocelis subcapitata, Ceriodaphnia dubia, or Parhyale hawaiensis. Additional biochemistry/molecular studies are needed to elucidate the toxic/mutagenic pathways of emodin in aquatic organisms. The PNEC value for emodin was 0.025 µg L-1. In addition to mutagenicity in the Salmonella/microsome assay, emodin was mutagenic in the micronucleus assay in the amphipod P. hawaiensis. Among the anthraquinone dyes tested to date, natural or synthetic, emodin was the most toxic to aquatic species.

Is natural better? An ecotoxicity study of anthraquinone dyes.

The concept of sustainability has gained prominence in recent years, enhancing the need to develop products that are less harmful to the environment. Dyes are used by various industrial sectors and have a lot of market value; they are used on a large scale mainly by the textile industry that uses large volumes of water and is one of the main contributors to the contamination of water bodies. Some natural compounds, especially anthraquinones are re-emerging as possible alternatives to synthetic dyes, some of which are known for their toxic and/or mutagenic effects. The BioColour project (https://biocolour.fi/) which is interested in promoting the development of new alternative molecules to synthetic dyes, provided us highly purified anthraquinone dyes dermocybin and dermorubin (>98% purity) extracted from a specie of fungus Cortinarius sanguineus. Dyes were tested for their acute and chronic toxicity using different aquatic organisms. Dermorubin was not toxic to any of the organisms tested for the highest test concentration of 1 mg L-1 and it was the most promising dye. Dermocybin was toxic to Daphnia similis (EC50 = 0.51 mg L-1), Ceriodaphnia dubia (IC10 = 0.13 mg L-1) and Danio rerio embryos (extrapolated LC50 = 2.44 mg L-1). A safety limit, i.e, predicted no-effect concentration (PNEC) of 0.0026 mg L-1 was derived based on the toxicity of dermocybin. The PNEC value can be used to provide hazard information for future application in commercial dyeing processes. Then, we compared the toxicity of dermocybin and dermorubin with ecotoxicity data available in the literature on other anthraquinone dyes of natural and synthetic origin. Some natural dyes can be as toxic as synthetic ones, or more toxic when chronic effects are considered. Despite natural dyes being used since centuries past, there are few ecotoxicological studies available. This study is designed to help develop a more comprehensive understanding of their toxicological properties.

Electrochemical oxidation of indanthrene blue dye in a filter-press flow reactor and toxicity analyses with Raphidocelis subcapitata and Lactuca sativa.

Alternative routes to degrade dyes are of crucial importance for the environment. Hence, we report the electrochemical removal of indanthrene blue by using a boron-doped diamond anode, focusing on the toxicity of the treated solutions. Different operational conditions were studied, such as current density (5, 10, and 20 mA cm-2) and electrolyte composition (Na2SO4, Na2CO3, and NaNO3). Besides, the pH was monitored throughout the experiment to consider its direct influence on the ecotoxicity effects. The highest electrochemical oxidation efficiency, measured as color removal, was seen in the 180 min condition of electrolysis in 0.033 M Na2SO4, applying 20 mA cm-2, resulting in a color removal of nearly 91% and 40.51 kWh m-3 of energy consumption. The toxicity towards Lactuca sativa depends solely on pH variations being indifferent to color removal. While the inhibition concentration (IC50) for Raphidocelis subcapitata increases 20% after treatment (in optimized conditions), suggesting that the byproducts are more toxic for this specific organism. Our data highlight the importance of analyzing the toxicity towards various organisms to understand the toxic effect of the treatment applied.

Biomass burning particles in the Brazilian Amazon region: Mutagenic effects of nitro and oxy-PAHs and assessment of health risks.

Emissions from burning of biomass in the Amazon region have adverse effects on the environment and human health. Herein, particulate matter (PM) emitted from biomass burning in the Amazon region during two different periods, namely intense and moderate, was investigated. This study focused on: i) organic characterization of nitro- and oxy-polycyclic aromatic hydrocarbons (PAHs); ii) assessment of the excess lifetime cancer risk (LCR); and iii) assessment of the in vitro mutagenic effects of extractable organic matter (EOM). Further, we compared the sensitivity of two mutagenicity tests: Salmonella/microsome test and cytokinesis-block micronucleus (CBMN) with human lung cells. Among the nitro-PAHs, 2-nitrofluoranthene, 7-nitrobenz[a]anthracene, 1-nitropyrene, and 3-nitrofluoranthene showed the highest concentrations, while among oxy-PAHs, 2-metylanthraquinone, benz[a]anthracene-7,12-dione, and 9,10-anthraquinone were the most abundant. The LCR calculated for nitro-PAH exposure during intense biomass burning period showed a major contribution of 6-nitrochrysene to human carcinogenic risk. The EOM from intense period was more mutagenic than that from moderate period for both TA98 and YG1041 Salmonella strains. The number of revertants for YG1041 was 5-50% higher than that for TA98, and the most intense responses were obtained in the absence of metabolic activation, suggesting that nitroaromatic compounds with direct-acting frameshift mutagenic activity are contributing to the DNA damage. Treatment of cells with non-cytotoxic doses of EOM resulted in an increase in micronuclei frequencies. The minimal effective dose showed that Salmonella/microsome test was considerably more sensitive in comparison with CBMN mainly for the intense burning period samples. This was the first study to assess the mutagenicity of EOM associated with PM collected in the Amazon region using Salmonella/microsome test. The presence of compounds with mutagenic effects, particularly nitro- and oxy-PAHs, and LCR values in the range of 10-5 indicate that the population is potentially exposed to an increased risk of DNA damage, mutation, and cancer.

Investigation of the mutagenic and genotoxic activities of LLL-3, a STAT3 inhibitor.

LLL-3, an anthracene derived compound, has been shown to be a promising therapeutic agent for the treatment of some kinds of cancer such as chronic myeloid leukemia and glioblastoma. However, no data regarding the toxic properties of this compound have yet been described in the literature. The present work aimed to investigate the mutagenic and genotoxic activities of LLL-3 using the TA97, TA98, TA100, TA102 and TA104 Salmonella/microsome strains for the Ames test and the micronucleus assay with the mouse macrophage cell line RAW 264.7. The findings showed that LLL-3, at doses of 0.001, 0.01, 0.1, 1.0 and 10.0 µg/plate, did not induce mutagenic activity in the Salmonella strains used under the conditions tested, and nor did it present genotoxicity in RAW 264.7 cells, at 10.0, 100.0 and 1000.0 µg/mL doses. Moreover, it is important to point out that the mitotic index of the cells decreased after exposure to LLL-3 under the same conditions tested, which may suggest some cytostatic effect, since this compound acts by inhibiting STAT3. Since most drugs used in the treatment of cancer present mutagenic activity as an adverse effect, these results suggest that LLL-3 is a promising drug for cancer therapy.

Eco- and genotoxicological assessments of two reactive textile dyes.

Contamination of natural waters has been one of the major problems of modern society and the textile industry is rated as an important polluting source, due to the generation of large amounts of wastewaters. The aim of this study was to assess textile dyes Reactive Blue 19 (RB19, anthraquinone dye) and Reactive Red 120 (RR120, azo dye) in terms of the potential to induce adverse effects on aquatic organisms and humans. Thus, these dyes were tested using the following assays: Microtox assay (Vibrio fischeri); brine shrimp (Artemia salina); Daphnia similis; and Comet with normal human dermal fibroblasts as well as Ames test (TA98, TA100, YG1041, YG1042--with and without S9). RB19 was relatively nontoxic to all aquatic bioindicators analyzed with an EC50 of more than 100 mg/L, whereas RR120 was only moderately toxic to A. salina with a EC50-48h of 81.89 mg/L. Mutagenicity through base pair substitution was observed with RB19 in the presence of S9 (Ames-positive). The comet assay did not demonstrate any apparent genotoxic effects for any tested dye. Although mutagenicity was detected with RB19, the mutagenic effect observed may be considered weak compared to the ability to induce DNA damage by other classes of dyes such as disperse dyes. Therefore, these dyes may be classified as nonmutagens (RR120) or weak mutagens (RB19) and relatively nontoxic for aquatic organisms. However, it is noteworthy that the weak acute toxicity to A. salina induced by RR120 is sufficient to suggest potential damage to the aquatic ecosystem and emphasizes the need for biomonitoring dye levels in wastewater systems.

Development and initial experience with a colored perfluorocarbon liquid for intraocular tamponade in vitreoretinal surgery.

PURPOSE: To present the development and initial experience of a novel colored perfluorocarbon liquid (PFCL) in vitreoretinal surgery. METHODS: This was an experimental laboratory study and prospective human interventional study. F6H8 (Fluoron GmbH) was colored by adding 0.3 g/L blue anthraquinone dye. Subsequently, 20% colored F6H8 was prepared by mixing with perfluorooctane or perfluorodecalin (Fluoron GmbH). The novel product is not yet FDA approved for human application. In the laboratory, the colored PFCL was covered with 1) uncolored PFCL, 2) BSS, and 3) silicone oil. Cell toxicity was evaluated in L929 mouse fibroblasts using a growth inhibition assay. Porcine ex vivo eyes were evaluated after vitrectomy followed by intravitreal and subretinal colored PFCL infusion. A pilot, prospective, noncomparative interventional study was conducted in patients with retinal detachment with proliferative vitreoretinopathy (PVR). RESULTS: The density of the colored PFLC mixture was 1.664 g/cm for perfluorooctane and 1.802 g/cm for perfluorodecalin. There was no relevant cell growth inhibition with any concentration of colored PFCL tested. Experiments in pigs revealed that infusion of the colored PFCL caused neither staining of the internal limiting membrane nor intravitreal residual droplets. In the prospective study, 9 eyes (75%) underwent surgery for rhegmatogenous retinal detachment with at least grade C PVR. The colored PFCL enabled retinal break examination and detection of residual intravitreal droplets in all surgeries. There was no case of separation or leakage of the dye from the PFCL solution that could have caused unwanted staining of the vitreous or epiretinal surface. CONCLUSION: The colored PFCL enabled intraoperative maneuvers such as endolaser use. In addition, removal of the colored PFCL was easily achieved at the end of surgery.

Synthesis and evaluation of cytotoxicity and inhibitory effect on nitric oxide production by J774A.1 macrophages of new anthraquinone derivatives.

Mitoxantrone is an anthracene-based anticancer agent whose efficacy in treating autoimmune diseases is believed to be due to cytotoxicity and inhibition of proliferation of cells. Several novel anthraquinone derivatives, analogs of mitoxantrone, were designed and synthesized. Lipophilic and functionalized mitoxantrone analogs were prepared by a simple methodology and the cytotoxicity and the inhibitory effect on nitric oxide release of these compounds were demonstrated in vitro on J774A.1 macrophages. Interestingly compounds 3, 4, 5, 6, 7, and 8 exhibited reduction in NO release (62.4%, 92.6%, 73.4%, 58.4%, 57.8% and 53.4%, respectively) in comparison to NG-n-methyl-arginine treated control, without cytotoxicity. In conclusion, anthraquinone derivatives were prepared in a good yield and showed promissory antiinflammatory properties.

In-vivo laxative and toxicological evaluation and in-vitro antitumour effects of Mitracarpus frigidus aerial parts.

OBJECTIVES: To evaluate the in-vitro antitumour properties, and the in-vivo laxative and toxicological effects of the methanolic extract of the aerial parts of Mitracarpus frigidus (MFM). METHODS: The in-vitro antitumour activity of MFM was evaluated against three human tumour cell lines: Jurkat, HL60 and MCF-7. The laxative activity and the effect of MFM on intestinal motility were evaluated in rats at the doses of 100, 300 and 1000 mg/kg. Acute oral toxicity was performed at 10, 100, 1000 and 2000 mg/kg and subchronic toxicity was evaluated at 100, 300 and 1000 mg/kg of MFM during a 42-day period. After subchronic administration of MFM the biochemical, haematological and histopathological parameters were analysed. Also, the total content of anthraquinones was determined. KEY FINDINGS: MFM was cytotoxic only against HL60 and Jurkat cells with 89 and 83% growth inhibition, respectively. The laxative activity of MFM was similar to bisacodyl. Regarding the effect on intestinal motility, MFM showed a significant increase in the pathway of charcoal compared with the group treated with saline. Furthermore, MFM showed no in-vivo toxicity at the doses tested. Free and anthraquinone C- and O-glycosides were detected in MFM. CONCLUSIONS: MFM showed significant antitumour activity for leukaemic cells. Moreover, it presented laxative potential and no in-vivo toxicity.

Phototoxic effects of Heterophyllaea pustulata (Rubiaceae).

Heterophyllaea pustulata (Rubiaceae), a South American genus, is a phototoxic shrub that grows in the Andean mountain range of the northwest of Argentina, popularly known as "cegadera". Animals that ingest the aerial parts of this plant suffer a typical primary photosensitization reaction, clinically revealed by dermatitis and blindness in severe cases. Anthraquinone derivatives (AQs), the main metabolites of this species, are characterized as Type I and/or Type II photosensitizers according to their physicochemical properties. The natural toxicity conditions were reproduced in vivo assays by oral administration of soranjidiol and rubiadin, the main components of the aerial parts. By HPLC analysis, the presence of these AQs was determined in serum and quantified in the skin of experimental animals.

New 1,2,3,4-tetrahydro-1-aza-anthraquinones and 2-aminoalkyl compounds from norlapachol with molluscicidal activity.

New nitrogen derivatives from norlapachol, including four new diastereomeric 1,2,3,4-tetrahydro-1-aza-anthraquinones obtained from the Prins cyclization on suitable aminoacetaldehyde dimethylacetal derivatives with formic acid, were found to exhibit molluscicidal activity against Biomphalaria glabrata. These derivatives showed low to medium LC(50) values, similar to those reported previously for the homologous series of nitrogen derivatives of lapachol. The toxicity profile against Artemia salina was also determined for all compounds.

Oncocalyxone A from Auxemma oncocalyx lacks genotoxic activity in phytohemagglutinin-stimulated lymphocytes.

Inadequate doses or prolonged chemotherapy can be cytotoxic or genotoxic to cancer patients, increasing the risk for the development of a second cancer, particularly acute leukemia. The association between therapeutic and genotoxic properties of oncocalyxone A (Onco A), make cytotoxic tests (mitotic index and chromosomal aberrations) fundamental in the accompaniment of the effects of this active compound. Therefore, the aim of the present study was to determine the genotoxic action of Onco A in vitro, during different phases of the cell cycle, utilizing primary cultures of lymphocytes of healthy individuals. The results showed that Onco A is cytotoxic during the cell cycle phases G1, G1/S, and S, however, not in G2. Onco A did not demonstrate a genotoxic effect in any of the cell cycle phases at the concentration studied. It is concluded that during the period of exposure, this active substance inhibits DNA synthesis and consequently cell division. Therefore, the absence of such genotoxicity for Onco A in the tests performed in this study provides important information in regard to the therapeutic use of this agent. Further studies are necessary to better understand the molecular mechanism of action of Onco A.

A degradation and toxicity study of three textile reactive dyes by ozone.

Textile reactive dyes are very recalcitrant to biodegradation, causing aesthetic, acute and chronic toxicity problems in receiving waters. In this study, we present the degradation of three different reactive dyes (Reactive Black 5, Reactive Blue 19 and Reactive Blue 21) by ozone, at two different pH conditions. The ozonation process achieved a very fast decoloration rate for the studied dyes. The influence of pH in the ozonation of these dyes was only significant for Reactive Black 5. The toxicity of dyes, after pre-treatment, was evaluated using Vibrio fisheri (microtox). There was little change in toxicity for Reactive Black 5, but there was a reduction for Reactive Blue 19. A significant increase in toxicity for Reactive Blue 21 was verified, caused by the increase in the bioavailability of th e cooper ion that was in the complexed form before the treatment with ozone.

Toxicity to sea urchin egg development of the quinone fraction obtained from Auxemma oncocalyx.

Auxemma oncocalyx Taub. belongs to the Boraginaceae family and is native to the Brazilian northeast where it is known as "pau-branco". We investigated the ability of the water soluble fraction isolated from the heartwood of A. oncocalyx to inhibit sea urchin egg development. This fraction contains about 80% oncocalyxone A (quinone fraction), a compound known to possess strong cytotoxic and antitumor activities. In fact, the quinone fraction inhibited cleavage in a dose-dependent manner [IC50 of 18.4 (12.4-27.2) microg/ml, N = 6], and destroyed the embryos in the blastula stage [IC50 of 16.2 (13.7-19.2) microg/ml, N = 6]. We suggest that this activity is due to the presence of oncocalyxone A. In fact, these quinones present in A. oncocalyx extract have strong toxicity related to their antimitotic activity.

Antiproliferative effects of compounds derived from plants of Northeast Brazil.

Ten compounds derived from plants indigenous to Northeast Brazil were examined for antiproliferative effects on human cells in vitro. The effects of these phytochemicals on cell growth were determined by the MTT microtitre assay with 3-day continuous drug exposure. Three human cell lines were used: CEM leukaemia, SW1573 lung tumour and CCD922 normal skin fibroblasts. Four active compounds were found with IC(50) values less than 10 microg/mL in the two cancer cell lines. Oncocalyxones A and C, both 1,4-anthracenediones from Auxemma oncocalyx (Boraginaceae), showed cytotoxicity with mean IC(50) values of 0.8-2, 7-8 and 12-13 microg/mL against CEM, SW1573 and CCD922, respectively. One diterpene and one flavonoid, both from Egletes viscosa (Compositae), were also active. 12-Acetoxy-hawtriwaic acid lactone was cytotoxic with mean IC(50) values of 6, 10 and 10 microg/mL, respectively. 4,5-Dihydroxy-3,3,7, 8-tetramethoxy flavone (ternatin) was only growth-inhibitory with mean IC(50) values of 2, 1 and 10 microg/mL, respectively. These four most active compounds were examined further for their effects on DNA integrity and on DNA synthesis. All but ternatin caused substantial DNA damage and marked inhibition of 5-bromo-2'-deoxyuridine incorporation within 24 h. This study demonstrated the antiproliferative activity of four novel phytochemicals, three of which are DNA-reactive and inhibit DNA synthesis. Further studies are warranted to evaluate these compounds for antitumour potential.