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Previous studies have shown that differences in experimental design factors may alter the potency of genotoxic compounds in in vitro genotoxicity tests. Most of these studies used traditional statistical methods based on the lowest observed genotoxic effect levels, whereas more appropriate methods, such as the benchmark dose (BMD) approach, are now available to compare genotoxic potencies under different test conditions. We therefore investigated the influence of two parameters, i.e. cell type and exposure duration, on the potencies of two known genotoxicants [aflatoxin B1 and ethyl methanesulfonate (EMS)] in the in vitro micronucleus (MN) assay and comet assay (CA). Both compounds were tested in the two assays using two cell types (i.e. CHO-K1 and TK6 cells). To evaluate the effect of exposure duration, the genotoxicity of EMS was assessed after 3 and 24 h of exposure. Results were analyzed using the BMD covariate approach, also referred to as BMD potency ranking, and the outcome was compared with that of more traditional statistical methods based on lowest observed genotoxic effect levels. When comparing the in vitro MN results obtained in both cell lines with the BMD covariate approach, a difference in potency was detected only when EMS exposures were conducted for 24 h, with TK6 cells being more sensitive. No difference was observed in the potency of both EMS and aflatoxin B1 in the in vitro CA using both cell lines. In contrast, EMS was more potent after 24 h exposure compared with a 3 h exposure under all tested conditions, i.e. in the in vitro MN assay and CA in both cell lines. Importantly, for several of the investigated factors, the BMD covariate method could not be used to confirm the differences in potencies detected with the traditional statistical methods, thus highlighting the need to evaluate the impact of experimental design factors with adequate approaches.

Persistent organic and inorganic contaminants generated by industrial effluent wastes poses a threat to the maintenance of aquatic ecosystems and public health. The Khniss and Hamdoun rivers, located in the central-east of Tunisia, receive regularly domestic and textile wastewater load. The present study aimed to survey the water quality of these rivers using physicochemical, analytical and toxicological approaches. In the physicochemical analysis, the recorded levels of COD and TSS in both samples exceed the Tunisian standards. Using the analytical approach, several metals and some textile dyes were detected. Indeed, 17 metals were detected in both samples in varying concentrations, which do not exceed the Tunisian standards. The sources of metals pollution can be of natural and anthropogenic origin. Three textile disperse dyes were detected with high levels compared to other studies: the disperse orange 37 was detected in the Khniss river with a concentration of 6.438 μg/L and the disperse red 1 and the disperse yellow 3 were detected in the Hamdoun river with concentrations of 3.873 μg/L and 1895 μg/L, respectively. Textile activities were the major sources of disperse dyes. For both samples, acute and chronic ecotoxicity was observed in all the studied organisms, however, no genotoxic activity was detected. The presence of metals and textile disperse dyes could be associated with the ecotoxicological effects observed in the river waters, in particular due to the industrial activity, a fact that could deteriorate the ecosystem and therefore threaten the human health of the population living in the study areas. Combining chemical and biological approaches, allowed the detection of water ecotoxicity in testing organisms and the identification of possible contributors to the toxicity observed in these multi-stressed water reservoirs.

Due to the complexity and variability of textile wastewater composition, a constant search for new treatment strategies that are efficient, eco-friendly, and cost-effective is mandatory. In the present study, the efficiency of coagulation-flocculation using biocoagulants derived from cactusOpuntia ficus indicaand eggplantSolanum melongenato remove toxic compounds from Tunisian textile wastewater samples was evaluated by combining assays to investigate physicochemical properties andin vitro(geno)toxicity with analytical chemistry. Both natural coagulants could significantly improve the physicochemical properties of the textile wastewater samples compared to the traditionally used chemical coagulant. The highest rate of decolorization was achieved after treatment with the cactus-derived coagulant. The analytical study revealed the presence of only crystal violet dye (CV) in only one sample. Both natural coagulants were able to remove CV, which may (partially) explain the decolorization of the treated samples. Only one untreated textile effluent induced a genotoxic response in the VITOTOX (R) assay. The genotoxic effect was not linked to the presence of CV and was no longer observed after treatment with each of the natural coagulants, suggesting the effectiveness of the remediation treatments to remove potentially genotoxic compound(s). However, in the other genotoxicity tests, no biologically relevant effects were observed for any of the tested samples. In conclusion, although the physicochemical data indicate that the use of natural coagulants (cactus and eggplant) could be an interesting alternative treatment process to the chemical coagulant for detoxifying textile effluents, these results were only partially supported by the toxicological and analytical data.

In the present study, a battery of biological tests undertaken in vitro and in vivo was used to evaluate the toxic potential of an organophosphorus insecticide, namely Fenitrothion. The cytotoxic effect of pesticide was evaluated with the MTT assay against two human cancer cell lines: Hep-2 and MDA-MB-231. Genotoxicity was also studied using the bacterial VITOTOX® assay. The estrogenic effect was tested using the recombinant yeasts (YES) assay. Likewise, bioluminescence assays using V. fischeri and D. magna immobilization were performed. The results showed that Fenitrothion exhibits a variable cytotoxic effect depending on the dose as well as the studied cell lines, and no genotoxicity was observed in the tested sample. However, an estrogenic effect was recorded when investigating Fenitrothion using the recombinant yeasts (YES) assay. Analogously, acute toxicity was observed for both organisms and at all tested concentrations of Fenitrothion. Overall, these results underline the crucial importance of in vitro and in vivo bioassays in monitoring toxicity of pesticides.

Aim: Despite its therapeutic value almost nothing is known about potential adverse health effects of Olea europea L. We therefore investigated the in vitro toxicity and genotoxicity of leaf extracts of this plant. Material and Methods: Extracts from olive tree leaves were obtained from four different regions in Tunisia. We investigated the in vitro toxicity, genotoxicity and antigenotoxicity of their aqueous extracts using the neutral red (NR) uptake, Vitotox and alkaline comet assays. Results: None of the extracts were found to be toxic and none of them were genotoxic, although some doubt exists for the extract obtained at Meski (North of Tunisia). On the basis of the Vitotox test only, none of the extracts appeared to have antigenotoxic (or cogenotoxic) properties. Discussion: The negative genotoxicity underline the safe use of the leaves, for example, as hypoglycemic and antidiabetic preparations. Lack of antigenotoxicity may indicate that the previously reported anticancer effects do not result from protection against genotoxicity. HIGHLIGHTS We investigated the in vitro toxicity and genotoxicity of aqueous extracts of olivesThe neutral Red Uptake test, Vitotox and alkaline comet assay were usedLeaf extracts from 4 different origins were investigatedNone of them showed in vitro toxicity or genotoxicityThe extracts also didn’t have antigenotoxic properties Abbreviation list: BaP: benzo(α)pyrene, EMS: ethyl methane sulfonate, LMP: low melting point, NI50: 50% inhibition of NRU, NR: neutral red, NRU: neutral red uptake, OD: optical density, PBS: phosphate buffer saline, SDS: sodium dodecyl sulphate, S/N: signal to noise ratio, 4NQO: 4-nitroquinoline oxide

Beet root (Beta vulgaris L.) is well known in cuisine and is also used in traditional medicine to prevent many diseases. We investigated the methanolic extract and juice with respect to the phenolic and flavonoid contents and evaluated their antioxidant, anticoagulant and genotoxic activities. Genotoxicity was studied with the bacterial Vitotox test and the alkaline comet assay involving human C3A liver cells. The methanolic extract had the highest phenolic and flavonoid contents with values of 39.75 mg GAE/g and 20.73 mg CE/g extract, respectively. Methanolic extract and juice showed surprising anticoagulant and antioxidant activities. They were devoid of any genotoxicity or cytotoxic effects. The results show the potential of the plant, especially the juice, as a new source of bioactive molecules for therapeutic purposes, possibly in the treatment of cardiovascular diseases. (C) 2019 SAAB. Published by Elsevier B.V. All rights reserved.

Nigella sativa or black cumin is used as a spice and as a natural remedy against a great variety of illnesses. However, plants growing at different locations, i.e., coping with different environmental stress conditions, may have different contents of specialized compounds include alkaloids, flavonoids, and terpenoids. Therefore their properties and biological effects may be different. We used the neutral red uptake test, the bacterial Vitotox and Ames assays and the micronucleus and comet tests in human C3A cells to investigate the in vitro cytotoxicity, genotoxicity and antigenotoxicity of aqueous seed extracts of this plant originating from three different regions in Morocco. Nigella sativa seed extracts showed different degrees of cytotoxicity depending on the location where the test samples came from. Overall they were not genotoxic but some indications of both antigenotoxicity and co-genotoxicity were found. In this the results are somewhat in contradiction with previous reports. Actually, our results show that many factors may influence the outcome of an investigation. The location of the samples, but also the applied test system and other experimental conditions are examples of factors that should be taken into account when evaluating the biological properties and potentials for nutritional and medicinal application in humans. This obviously complicates these kind of investigations and assessment and shows that it may be time to develop and agree on common protocols and procedures rather than performing experiments solely based on a laboratory's capacity, availabilities and opportunities. (C) 2019 SAAB. Published by Elsevier B.V. All rights reserved.

E-cigarettes have become very popular, a trend that has been stimulated by the wide variety of available e-liquid flavours. Considering the large number of e-liquid flavours (>7000), there is an urgent need to establish a screening strategy to prioritize the flavouring substances of highest concern for human health. In the present study, a prioritization strategy combining analytical screening, in silico tools and literature data was developed to identify potentially genotoxic e-liquid flavourings. Based on the analysis of 129 e-liquids collected on the Belgian market, 60 flavourings with positive in silico predictions for genotoxicity were identified. By using literature data, genotoxicity was excluded for 33 of them whereas for 5, i.e. estragole, safrole, 2-furylmethylketon, 2,5-dimethyl-4-hydroxyl-3(2H)-furanone and transhexanal, there was a clear concern for in vivo genotoxicity. A selection of 4 out of the remaining 22 flavourings was tested in two in vitro genotoxicity assays. Three out of the four tested flavourings induced gene mutations and chromosome damage in vitro, whereas equivocal results were obtained for the fourth compound. Thus, although there is a legislative framework which excludes the use of CMR compounds in e-liquids, flavourings of genotoxic concern are present and might pose a health risk for e-cigarette users.

Dictyopteris membranacea, a species of Mediterranean brown algae, is believed to have potential pharmacological and nutritional applications. However, such potentials only make sense when devoid of any adverse health consequences. The present study should be seen in this context. It aimed at evaluating the genotoxicity and cytoxicity of its organic extract (F0) and semi purified fractions (F4, F5, and F6). Extracts were tested using the bacterial Vitotox® test and micronucleus assay in different concentrations (from 1.25 μg/mL up to 100 μg/mL, depending on the test and the extract). Applied concentrations were based on a preliminary dose-finding test with the neutral red uptake assay. The results show that all extracts were not genotoxic in the presence or absence of a rat metabolic enzyme fraction (S9). This is encouraging and justifies further investigations on the therapeutic and other values of this algae. SUMMARY Dictyopteris membranacea extracts and some of their semi purified fractions have important antibacterial properties.The organic extract (F0) and semi purified fractions (F4, F5, and F6) were not genotoxic according to the bacterial Vitotox test.They were also not genotoxic according to the micronucleus test in human C3A cells.Applied concentrations were based on the in-vitro neutral red uptake (NRU) test.

Potential health effects of extremely low-frequency (electro)magnetic fields (ELF-(E)MFs) have long been investigated, but the results are still inconclusive. With respect to genotoxicity, sound data related to the effects of long-term exposure to ELF-(E)MFs on the genetic material and the impact of long-term pre-exposure to ELF-(E)MFs on the sensitivity of cells to the damage induced by known mutagens are needed. In this manuscript, an optimized protocol for a combined in vitro comet/micronucleus study to investigate these effects in a human lymphoblastoid cell line (TK6) is provided including the description of a well-validated exposure system. Furthermore, the use of a shielding system to limit background ELF-MFs inside the incubator is described as well. • Optimized protocols for cytogenetic tests with ELF-MFs on TK6 cells ensure the reproducibility of test results. • Validation of exposure environment and exposure system are needed prior to performing tests with ELF-MFs. • A simple, but effective method to shield cells and reduce unintentional ELF-MF exposure consists of using the mu-metal cylinder. This is of particular interest when studying the effects of low exposure levels., SCOPUS: ar.j, info:eu-repo/semantics/published

Ruscus hypophyllum L. is a rare Mediterranean plant which is used in the traditional medicine. We studied its phenolic content and in vitro toxicity and genotoxicity using the neutral red uptake (NRU) test, the bacterial Vitotox test, and the comet assay in human C3A hepatic cells. Aqueous leaf and fruit extracts were investigated. Antigenotoxicity against 4-nitroquinoline-oxide (4NQO, 0.4 mu g/mL) and Benzo(alpha) pyrene (BaP, 800 mu g/mL) was also investigated with the Vitotox test. The extracts appeared to be genotoxic only at high exposure levels in the comet assay. There was no indication of a genotoxic activity in the Vitotox test and also no indication of antigenotoxicity. The moderate polyphenol content may provide an explanation for the absence of antigenotoxicity.

The UPLC MS/MS analysis showed the presence of the two antibiotics in the pharmaceutical industry discharges during 3 months; norfloxacin and spiramycin which were quantified with the mean concentrations of 226.7 and 84.2 ng mL(-1), respectively. Sixteen resistant isolates were obtained from the pharmaceutical effluent and identified by sequencing. These isolates belong to different genera, namely Citrobacter, Acinetobacter, Pseudomonas, Delftia, Shewanella, and Rheinheimera. The antibiotic resistance phenotypes of these isolates were determined (27 tested antibiotics-discs). All the studied isolates were found resistant to amoxicillin and gentamicin, and 83.33% of isolates were resistant to ciprofloxacin. Multiple antibiotic resistances were revealed against beta-lactams, quinolones, and aminoglycosides families. Our overall results suggest that the obtained bacterial isolates may constitute potential candidates for bioremediation and can be useful for biotechnological applications. Genotoxic effects were assessed by a battery of biotests; the pharmaceutical wastewater was genotoxic according to the bacterial Vitotox test and micronuclei test. Genotoxicity was also evaluated by the comet test; the tail DNA damages reached 38 and 22% for concentrated sample (10x) and non-concentrated sample (1x), respectively. However, the histological sections of kidney and liver's mice treated by pharmaceutical effluent showed normal histology and no visible structural effects or alterations as cytolysis, edema, or ulcerative necrosis were observed. Residual antibiotics can reach water environment through wastewater and provoke dissemination of the antibiotics resistance and induce genotoxic effects.

A series of prenylated flavonoids was obtained from antigenotoxic extracts and fractions of stem bark of Erythrina latissima E. Mey (Leguminosae). In addition to five constituents never reported before, i.e. (2 S )-5,7-dihydroxy-2-(4-hydroxy-2-(prop-1-en-2-yl)-2,3-dihydrobenzofuran-6-yl)chroman-4-one (erylatissin D), (2 S )-5,7-dihydroxy-2-(4-methoxy-2-(prop-1-en-2-yl)-2,3-dihydrobenzofuran-6-yl)chroman-4-one (erylatissin E), 5,7-dihydroxy-3-(4-methoxy-2-(prop-1-en-2-yl)-2,3-dihydrobenzofuran-6-yl)-4H-chromen-4-one (erylatissin F), (2 S )-5,7,8′-trihydroxy-2′,2′-dimethyl-[2,6′-bichroman]-4-one (erylatissin G) and (2 S )-5,7-dihydroxy-8′-methoxy-2′,2′-dimethyl-[2,6′-bichroman]-4-one (dihydroabyssinin I), 18 known flavonoids were identified. Evaluation of the antigenotoxic properties (against genotoxicity induced by aflatoxin B1, metabolically activated) in the Vitotox assay revealed that most flavonoids were active. Sigmoidin A and B showed the highest activity, with an IC 50 value of 18.7 μg/mL, equivalent to that of curcumin (IC 50 18.4 μg/mL), used as a reference antigenotoxic compound.

The emergence and increase of microbial resistance towards many conventional therapeutics necessitate the investigation and discovery of new natural sources of antimicrobial agents. In this study the Mediterranean brown alga, Dictyopteris membranacea, was investigated. The total phenolic, flavonoid and tannin contents were quantified and antibacterial, antifungal and antitubercular activities of the crude extract and semi-purified ethanol, acetone and methanol/CH2Cl2 fractions were determined. The antimicrobial activity was determined by the agar diffusion method against six strains of Gram-positive, two strains of Gram-negative bacteria and one yeast strain. The MIC and MBC or MFC values served to identify the bactericidal and bacteriostatic effects of all prepared fractions against the different strains. The acetone fraction presented a source of phenolic (112 mg GAE/g of sample) and tannin (130 mg CAE/g sample) compounds whereas the ethanol fraction showed the highest concentration of flavonoids (120 mg QE/g). Ethanol and acetone fractions exhibited considerable antibacterial, antifungal and antitubercular activities with a good correlation between the phytochemical composition of all prepared fractions and the different biological activities. This study on the phytochemical and in vitro antimicrobial activity of this brown alga was supplemented by an in vitro investigation of the potential genotoxicity of the extract and fractions using the bacterial Vitotox test and micronucleus test on human C3A cells. The results show that all prepared fractions were not genotoxic in the presence or absence of S9. Overall, the results are therefore encouraging and justify further investigations on the therapeutic and other values of this alga. (C) 2016 SAAB. Published by Elsevier B.V. All rights reserved.

In the following study, we came up with and validated a prompt, sensitive and precise method for the simultaneous determination of 56 antimicrobial drugs (tetracyclines, sulfonamides, β-lactams, macrolides and quinolones) using the ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). This method was implemented with success to determine antibiotics in samples collected from four wastewater treatment plants and five coasts in Tunisia. Results showed the presence of high concentrations of antibiotics that ranged from 0.1 to 646 ng mL−1, which can induce many negative impacts on health and the environment. For this reason, we have opted to evaluate toxicity of wastewater samples using a battery of biotests. In fact, genotoxicity was assessed using three tests: Vitotox, comet and micronucleus assays. The input and output of wastewater treatment plants induced a strongly genotoxic effect on the Salmonella typhimurium TA104 prokaryotic Vitotox assay. This result was confirmed using the comet and the micronucleus assays performed on the human liver hepatocellular carcinoma cells. The genotoxic power of the tested wastewater treatment plants' samples could be attributed to the presence of the higher quantities of antibiotics that are detected in these samples and to the antibiotic and organic compound cocktails.

Tuberculosis (TB) continues to be a worldwide health problem with over 1.4 million deaths each year. Despite efforts to develop more effective vaccines, more reliable diagnostics, and chemotherapeutics, tuberculosis remains a threat to global health, fueled by the HIV pandemic and the rapid generation of drug resistance. The exploration of novel drugs to serve as a companion drug for existing drugs is of paramount importance. As part of our program to design new 2-aza-anthraquinones with antimycobacterial activity, various tetrahydro- and octahydrobenzo[j]phenanthridinediones were synthesized. These compounds showed high in vitro potency against Mycobacterium tuberculosis, the etiological agent of TB and against other clinically relevant mycobacterial species at submicromolar concentrations. The susceptibility of a multidrug resistant strain toward these compounds and their ability to target intracellular replicating Mycobacterium tuberculosis was demonstrated. Next to the acute toxicity, the genotoxicity of these compounds was investigated. Often overlooked in studies, genotoxicity could be dismissed for the investigated compounds, making them a promising scaffold in TB drug research.

There are a few publications in scientific journals suggesting that persons who are regularly exposed to ELF-magnetic fields (occupational-, but also residential exposures) may be at risk for contracting Alzheimer's disease. It should be noted that these reports are only pointing to a possible association but do not at present provide any proof of association. We have performed a number of preliminary in vitro laboratory investigations to explore the plausibility of such an association. Our investigations were based on similarities found in cells from Alzheimer disease patients and in cells exposed to extreme low frequency magnetic fields. We especially investigated genetic damage in C3A and SH-SY5Y cells exposed in vitro to 50 Hz magnetic fields (0-500 mu T) using the cytome assay. We found an increased frequency of large micronuclei (suggesting a possible aneugenic effect) and increased frequency of nuclear buds (suggesting gene amplification). In this our results may show some similarities with observed micronuclei in blood and buccal cells of Alzheimer disease patients. The effects were found in both cell lines and were already observed for exposure levels as low as 10 mu T. This is however in contrast with earlier results obtained in human lymphocytes where no induction of micronuclei was found using the same exposure unit. The results thus need to be confirmed and further analyses need to be done.

It is believed that extreme low frequency magnetic fields (ELF-MF) are not mutagenic, at least at exposure levels below 100 mu T. Synergistic or co-operative effects with environmental mutagens remain possible yet. We therefore investigated the effects of ELF-MF in conjunction with 4 different well known chemical mutagens having different modes of action. For this purpose the bacterial Vitotox test was used. Our study confirmed previous results which showed that a 100 mu T magnetic field (50 Hz) does not damage DNA and hence is not mutagenic in this assay and that there was also no influence on the DNA damaging capacity of the used mutagens. (C) 2015 Elsevier B.V. All rights reserved.

Exposure to extremely low frequency magnetic fields (ELF-MF) has been identified as one of the potential environmental risk factors for Alzheimer's disease (AD). However, this is far from being established. So far there is no experimental evidence supporting this alleged association. We have performed an in vitro cytogenetic laboratory investigation to explore the plausibility of such association. Our investigation was based on possible similarities found in cells from AD patients and in cells exposed to ELF-MF. We especially found that 50 Hz ELF-MF increase the frequency of cells with (large) micronuclei and nuclear buds indicating that fields above 50 mu T may induce chromosome instabilities as those found in AD patients. It should be stressed yet that results from the few published experimental studies on ELF-MF and AD are rather reassuring. Thus, our findings certainly do not prove anything. They only suggest that further investigations might be necessary.

We performed an in vitro evaluation of the genotoxic potential of water extracts from four Hypoxis species (Hypoxis hemerocallidea, H. colchicifolia, H. rigidula, H. acuminata) and a commercial preparation thereof using the neutral red uptake (NRU) assay, the alkaline comet assay and the cytome assay in human hepatoma HepG2 cells. The relative cytotoxicity of these samples was established by determining their NI50 values (50% inhibition of NRU), and these results were used for dose-finding in genotoxicity tests. None of the tested extracts were identified as genotoxic in both the alkaline comet assay and cytome assay. Copyright © 2012 John Wiley & Sons, Ltd.

We investigated the genotoxic properties of a number of extracts from Tunisian traditional medicinal plants with the bacterial VITOTOX test in Salmonella typhimurium and the alkaline comet assay in human C3A cells. Ethyl acetate and methanol extracts from Marrubium alysson L. and Retama raetam (Forsk.) Webb and methanol extracts from Peganum harmala L. were investigated. Toxicity was furthermore studied with the neutral red uptake test that served for dose-finding. All extracts showed antigenotoxic properties against 4-nitroquinoline-oxide (4-NQO) and benzo(α)pyrene in the VITOTOX test, except the methanol extracts from R. raetam where antigenotoxicity was not found against the mutagen 4-NQO (in the absence of S9). The ethyl acetate extract from R. raetam was found mutagenic with the VITOTOX test in the absence of S9, whereas both ethylacetate and methanol extracts of M. alysson L. induced DNA damage according to the alkaline comet assay in C3A cells.

We performed a genotoxicity investigation of extremely low-frequency (ELF) magnetic fields (MFs, 50 Hz, 100 and 500 µT, 1 and 2 h exposure) alone and in combination with known chemical mutagens using the VITOTOX test. This test is a very sensitive reporter assay of Salmonella typhimurium bacteria based on the SOS response. Our study showed that ELF-MFs do not induce SOS-based mutagenicity in S. typhimurium bacteria and do not show any synergetic effect when combined with chemical mutagens.

Gene delivery has become an increasingly important strategy for treating a variety of human diseases, including infections, genetic disorders and tumours. To avoid the difficulties of using viral carriers, more and more non-viral gene delivery nanoparticles are developed. Among these new approaches polyethylene imine (PEI) is currently considered as one of the most effective polymer based method solution and considered as the gold standard. The toxicity of nanoparticles is a major concern when used for medical application. In this study we chose two nanoparticles for an in depth toxicological and ecotoxicological evaluation, one well characterized, PEI, and another novel polymer, poly(2-(dimethylamino)ethyl methacrylate) (PDMAEMA). In the present study we have assessed the toxicity of these cation nanoparticles as such and of the polyplexes nanoparticles covered with DNA. As these nanoparticles are also frequently used in high volumes in various industries and as such may enter in the environment, we also made an initial assessment of ecotoxicological effects assessment. The following nanoparticles related aspects have been studied during the project: development and characterization, ecotoxicity, general toxicity and specific toxicity. To this end a battery of different tests was used. The conclusion of these tests is that toxicity is varying between different nanoparticles and between different DNA covering ratios. In general, in the different systems tested, the PEI polymer is more toxic than the PDMAEMA polymer. The same difference is seen for the polyplexes and the higher the charge ratio, the more toxic are the polyplexes. Our study also clearly shows the need for a broad spectrum of toxicity assays for a comprehensive risk assessment. Our study has performed such a comprehensive analysis of two biomedical nanoparticles.

Cytotoxic and mutagenic effects of thirteen commercial herbal mixtures sold in KwaZulu-Natal, South Africa were evaluated using the neutral red uptake (NRU) assay and the Ames test. The herbal mixtures tested included Umzimba omubi , Umuthi wekukhwehlela ne zilonda , Mvusa ukunzi , Umpatisa inkosi , Imbiza ephuzwato , Vusa umzimba , Ingwe ® muthi mixture , Ibhubezi ™, Supreme one hundred ™, Sejeso herbal mixture Ingwe ®, Lion izifozonke Ingwe ®, Stameta ™ BODicare ® and Ingwe ® special muti . The relative cytotoxicity of the herbal mixtures was established by determining their NI 50 values (50% inhibition of neutral red uptake). The test revealed that the most toxic herbal mixture was Umpatisa inkosi with an NI 50 value of 0.016 mg/mL and the least toxic mixture was Stameta™ BODicare® with an NI 50 value of 28.00 mg/mL. The herbal mixtures showed no mutagenic effects against Salmonella typhimurium tester strains TA98, TA100, TA102, TA1535 and TA1537 when the assay was done without S9 metabolic activation. However, four herbal mixtures, Umpatisa inkosi , Imbiza ephuzwato , Vusa umzimba and Stameta ™ BODicare ® showed mutagenic effects against TA98 but not the rest of the tester strains after using S9 metabolic activation. Umpatisa inkosi also exhibited weak mutagenic activity against TA1535 after metabolic activation. The remaining mixtures did not show mutagenic effects against all the tester strains after S9 metabolic activation. The cytotoxic and mutagenic results reported here offer a step toward determining the safety of commercial herbal mixtures in South Africa. Herbal mixtures showing higher cytotoxic and mutagenic effects need to be further investigated for their possible effects on humans.

The main aim of the ACuteTox project (part of the EU 6th Framework programme) is to demonstrate that animal tests for acute systemic toxicity can be replaced by alternative in vitro assays. In this project, data for 97 reference chemicals were collected in the AcuBase database, designed to handle deposited in vitro and in vivo (human and animal) data. To demonstrate the applicability of in vitro basal cytotoxicity tests and in vitro– in vivo modelling, it was deemed necessary to obtain data that were generated via defined standard operating procedures. The molar basal cytotoxicity IC50 values (the 50% inhibitory concentrations for the endpoint measured) for a mouse fibroblast cell line (3T3), a human hepatic cell line (HepG2), a rat hepatic cell line (Fa32), and a human neutrophil cell line (HL-60), were compared, and gave an R2 correlation of 0.83. To identify chemicals that showed differential cytotoxicity to the various cell types involved, principal component analysis (PCA) was undertaken independently, once all the results had been returned. This showed that colchicine, cycloheximide, digoxin, 5-fluorouracil and hexachlorobenzene gave the lowest correlations with the first score vector of the PCA. The results presented are to be used to identify outliers that need to be further studied via the use of tissue-specific in vitro assays.

Plant-derived smoke and certain smoke compounds improve seed germination and enhance seedling growth of many species. Thus, smoke-infused water and the active smoke-derived compounds have the potential to be used in different agricultural and horticultural applications. However, despite these interesting and potentially practical properties, it should also be ascertained whether such compounds may pose a health risk, particularly if they are to be used in the production of food or fodder crops. Amongst some of the aspects that would be important to understand are any possible genotoxic properties that the compounds may possess due to potential carry-over effects. Here, we report on a genotoxicity study of 3,4,5-trimethylfuran-2(5H)-one, a compound from plant-derived smoke previously shown to have germination inhibitory activity. Using two in vitro tests, namely the bacterial VITOTOX® test (with/without S9 metabolic activation) and the cytome assay on human C3A cells, no genotoxicity or toxicity was found. Furthermore, these results support a previous study where a related smoke-derived compound with germination promoting properties was investigated.

In 1991, WHO recognized the resurgence of tuberculosis as a global health problem. Although modern chemotherapy is effective against the causative pathogen Mycobacterium tuberculosis, the current drug regimens have failed to eradicate the disease. The success of the pathogen, partially attributed to drug resistance, necessitates the development of novel anti-tuberculosis drugs. Benzo[j]phenanthridine-7,12-diones, tetracyclic derivatives of the natural product benz[g]isoquinoline-5,10-dione, were conveniently synthesized via palladium-catalyzed intramolecular cyclization of N-methanesulfonyl-3-bromo-2-(arylamino)methyl-1,4-naphthoquinones. Here we report on the bioactivity of eight benzo[j]phenanthridine-7,12-dione derivatives as candidate drug molecules against M. tuberculosis and on their cytotoxicity on C3A human hepatocytes. The strongest antimicrobial activity (as detected by growth inhibition of bacteria, using luminometry and BACTEC 460-TB) and lowest cytotoxicity was found for 3-methylbenzo[j]phenanthridine-7,12-dione 5e, which was also effective in targeting intracellular M. tuberculosis (in murine J774 macrophages) and was not genotoxic for C3A hepatocytes.

We conducted an investigation of the in vitro toxic and genotoxic properties of Cleome amblyocarpa Barr. and Murb, which is an important medicinal plant in Tunisia where it is widely used against colic and diabetes. In this study we investigated methanol extracts of the whole plant with the neutral red uptake test for assessing its in vitro toxicity and the alkaline comet assay for genotoxicity. It appeared that the extract was not genotoxic in subtoxic concentrations. However, the neutral red uptake test revealed that it was highly toxic and therefore some caution, especially regarding dosage and frequency of use, is warranted.