Investigation of the Mutagenic Potential of Immunomodulator Arglabin Native in Tablets in the Ames Test

The mandatory preclinical safety evaluation is an essential prerequisite to obtain the qualitative and effective medicines. Due to the fact that drugs may reveal genotoxic properties, the investigation of their mutagenic activity is an obligatory part of the preclinical drug safety program. The aim of the research is to study mutagenic properties of a new original immunomodulator Arglabin native in tablets in the induced test of gene mutations （the Ames test） on Salmonella typhimurium strains. Materials and methods： Four strains of S. typhimurium TA98, TA100, TA1535, and TA1537 were used to assess the mutagenicity in the Ames test. Results and conclusions： No statistically reliable dose-dependent increase in the number of revertant colonies of Salmonella typhimurium has been observed in the presence of the given drug within the investigated dose ranges from 5.0 to 100.0 μg/mL for strains TA100 and TA1535, and from 5.0 to 250 μg/mL for strains TA98 and TA1537 against the baseline of spantaneous mutations. Arglabin native in tablets does not reveal a mutagenic activity within the studied dose ranges on Salmonella typhimurium strains TA98, TA100, TA1535, TA1537.

Determination of Genotoxic Pollution of Some Hospital Wastewater with Salmonella Ames Test

Wastewater of hospitals contains materials that would be a threat to alive. These water needs to be checked by a biological purification before leaving to nature. Hospital wastewater has differences than domestic waste because of especially blood, body waste, drugs, chemicals, medical device waste and radioactive materials. We aimed to determine genotoxic effects of total pollution in hospital wastewater on alive by Salmonella microsome test method. In this study, we decided on three hospitals which weren’t checked as biological purification of waste. The samples were taken for six 1-week periods between March 2009 and June 2009. Mutagenite studies of samples taken from hospitals were made with ,Salmonella typhimurium TA 98 and TA 100. Wastewater samples were evaporated. 27 different test materials were prepared using DMSO, ethanol and acetone solvents, two different MGA plaques were used for each test material. Each experiment was made for 3 times with known results of mutagens and we made it ready for “Ames” test method. We had genotoxicity 50% in Istanbul University Medical Faculty Hospital, 56% in Haseki Hospital and 61% in Vak?f Gureba Hospital. According to three hospitals result there are 9 positives, 9 negatives in DMSO;9 positives, 9 negatives in ethanol;12 positives, 6 negatives in acetone. These values are totally 56%. Our results give important information about mutagenic effect of total pollution in hospital wastewater. It is first time researched in Turkey that effect on DNA of pollution is from hospital wastewaters. In prospective studies, it is necessary to use this system as a method to monitor mutagenic genotoxic pollution in hospital wastewaters. These kinds of studies present applicability and importance of our method because of placing in the literature. Method constitutes a new approach to check mutagenite of pollution in hospital wastewater.

Potential neoplastic effects of parathion-methyl on rat liver

The mutagenic and carcinogenic effects of parathion-methyl were examined by bacterial reverse mutation assay and a long-term experiment with wistar rats. The potential mutagenic effect of parathion-methyl in Salmonella typhimurium TA100 bacterial cells was observed without rat liver S9 metabolic activation. Parathion-methyl was further investigated for pathological changes in rat pancreas and liver. The long-term rat experiments showed that parathion-methyl exposure for 3 months can cause pathological changes in rat pancreases acinar cells and pancreatic hepatocytes. Atypical acinar cell focuses （AACF） were determined in the liver and pancreas of the rats. The results from short-term Ames test and long-term rat experiments suggested that parathion-methyl would be potential carcinogenic.

Study on the water quality of the Taihu Lake using genotoxicological methods

The micronucleus(MCN) test of Vicia faba root tip cell was used to screen the water quality of the Taihu Lake. The MCN ‰ and the pollution index ( PI ) were examined and the F test was carried out to evaluate the statistical difference in mean MCN ‰ among various samples. There were significant difference among thirty nine samples and there were six sites whose PI values are above 2. The authors suggest that the Taihu Lake has partly been polluted to different degrees and the whole water body can be divided into three types. Ames tests were then conducted to detect mutagenicity at five significant sites. The results showed that rivers flowing through urban area carried large amount of mutagenic pollutants into the lake and these pollutants contaminated the source of main waterworks.

In vitro Safety Evaluation and Anticlastogenic Effect of BacoMind^(TM) on Human Lymphocytes

Objective BacoMind^TM （BM） is a standardized extract of Bacopa monnieri, which belongs to the family Scrophulariaceae and is a creeping annual plant found throughout the Indian subcontinent. It has been used by Ayurvedic medicinal practitioners in India for almost 3000 years and is classified as a medharasayana, a substance which improves memory and intellect. With the widespread traditional use as well as scientific validation of Bacopa monnieri for nootropic activity, a bioactive-rich unique phytochemical composition-BacoMind^TM was developed from B. monnieri for use as a cognition and memory enhancing agent. The present study aimed to investigate the in vitro toxicity of this formulation of BacoMind^TM on human lymphocytes and to rule out its possible contribution to mutagenicity. Methods In the present investigation the active ingredients present in BM were identified and quantified by high performance liquid chromatography （HPLC） and high performance thin-layer chromatography （HPTLC）. Antioxidant and anticlastogenic properties of BM were studied in vitro with and without metabolic activation. Doses of BM were chosen on the basis of mitotic index （MI） and cytokinesis-block proliferation index （CBPI）. Clastogenicity assays were performed at 31.2 μg/mL, 62.5 μg/mL, and 125 μg/mL, while the Salmonella reverse mutation assay （Ames test） was performed at doses of 61.72, 185.18, 555.55, 1666.67, and 5000.00 μg/plate. Results HPLC and HPTLC analysis of BM revealed the presence of bacoside A3. bacopaside Ⅰ, bacopaside Ⅱ, jujubogenin isomer of bacopasaponin C, bacosine, luteolin, apigenin, bacosine, and β-sitosterol D glucoside. BM demonstrated significant antioxidant activity. The number of chromosomal aberrations and the frequency of micronuclei induced by BM were not statistically significant up to a dose of 62.5 μg/mL. A subsequent dose of 125 μg/mL prior to metabolic activation induced mild clastogenicity, but it was found to be biologically insignificant as this effect was not seen post metabolic activation. BM also demonstrated a dose-dependent protection against the clastogens used in this study using the above tests for clastogenicity. Maximum protection was observed in presence of metabolic activation. Moreover, BM demonstrated no mutagenic effect on the tested strains, as observed in the Ames test. Conclusion BM protected human lymphocytes against various clastogens. BM also exhibited high antioxidant activity which might be responsible for the observed protective effects against the clastogens since the used clastogens are known to induce their clastogenic effects via production of oxidative radicals.

Mutagenic and Estrogenic Effects of Organic Compounds in Water Treated by Different Processes: A Pilot Study

Objective In this study, a pilot-scale investigation was conducted to examine and compare the biotoxicity of the organic compounds in effluents from five treatment processes (P1-P5) where each process was combination of preoxidation (O3), coagulation, sedimentation, sand filtration, ozonation, granular activated carbon, biological activated carbon and chlorination (NaClO). Methods Organic compounds were extracted by XAD-2 resins and eluted with acetone and dichlormethane (DCM). The eluents were evaporated and redissolved with DMSO or DCM. The mutagenicity and estrogenicity of the extracts were assayed with the Ames test and yeast estrogen screen (YES assay), respectively. The organic compounds were detected by GC-MS. Results The results indicated that the mutation ratio (MR) of organic compounds in source water was higher than that for treated water. GC-MS showed that more than 48 organic compounds were identified in all samples and that treated water had significantly fewer types and concentrations of organic compounds than source water. Conclusion To different extents, all water treatment processes could reduce both the mutagenicity and estrogenicity, relative to source water. P2, P3, and P5 reduced mutagenicity more effectively, while P1 reduced estrogenicity, most effectively. Water treatment processes in this pilot plant had weak abilities to remove Di-n-butyl phthalate or 1, 2-Benzene dicarboxylic acid.

Synthesis and Antimutagenic Effects of Selena Morpholine Hydrate

The synthesis of selenamorpholine hydrate was described and its antimutagenic effects were investigated by the Ames tests. The results indicated that the mutation induced by the indirect mutagen 2-AF was inhibited obviously by selenamorpholine hydrate at the dosage of Se 0. 16--100 ug /dish for 35. 8% -53. 7% (TA_97) and the mutation induced by direct mutagen Dexon was inhibited for 6. 2% --30. 0% (TA_97,) and 2. 3% ~34. 1% (TA_100). The mutagenesis of the indirect mutagen cyclophosphamide was suppressed for 5.4%~ 16. 1% by selenamorpholine hydrate at the dosage of Se 0. 16--100 ug /dish.

Antioxidant and Antigenotoxic Activity of Bioactive Extracts from Corn Tassel

This study is designed to evaluate antioxidant and antigenotoxic activities of corn tassel extracts（CTTs）. The major bioactive components of CTTs include flavonoid, saponin and polysaccharide. The antioxidant properties of the three bioactive components of CTTs were investigated by Ferric Reducing Antioxidant Property（FRAP） and 1, 1-diphenyl-2-picrylhydrazyl（DPPH） assays. The activities of the extracts were determined by assessing the inhibition of mutagenicity of the direct-acting mutagen fenaminosulf, sodium azide, and indirect-acting mutagen 2-aminofluorene using the Ames test（strains TA98 and TA100）. The results showed that the extraction rates of flavonoid, saponin, and polysaccharide from the dried corn tassels were 1.67%, 2.41% and 4.76% respectively. DPPH and FRAP assay strongly demonstrated that CTTs had antioxidant properties. CTTs at doses of 625, 1250 and 2500 μg per plate reduced 2-aminofluorene mutagenicity by 12.52%, 28.76% and 36.49% in Salmonella typhimurium TA98 strain assay respectively and by 10.98%, 25.27% and 37.83%, at the same doses in Salmonella typhimurium TA100 assay system, respectively. 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide（MTT） assay showed that the different concentrations of CTTs inhibited the proliferation of MGC80-3 cells in a dose-dependent manner（P0.01）. It is concluded that these integrated approaches to antioxidant and antigenotoxicity assessment may be useful to study corn tassel as a natural herbal material.

Mutagenicity Study on Ibopamine: A New Heart Failure Antagonist

Ibopamine(Ibo) is a new effective drug against heart failure. This paper reports the results of mutation tests of chromosome aberration of mammalian cell (Chinese hamster lung cell, CHL) in vitro and reversional mutation test of Salmonella typhimurium bacterium (Ames test) of ibopamine . The results of both tests were negative. Ibo did not have mutagenic effect.

Chitosan that Dissolves in Hard Water Represses Mutagenicity

Chitosan is derived by deacetylation of chitin, a major component of the shells of crustacea such as crabs and shrimps. In this experiment using the Ames test, there was antimutagenic effect by the water-soluble chitosan that dissolved in hard water （CHW）. The Ames test was performed using Salmonella typhimurium TA98 and TA100. For the antimutagenic test, and the desmutagenic and bioantimutagenic tests, Trp-P-2, 2AA and AF-2 were used as mutagens. Tests for spontaneous mutation rate without mutagens were performed using S. typhimurium TA98 and TA100. When several concentrations of CHW were directly added to Minimal Glucose Plates, there were no changes in the number of His＋ revertants of TA98 and TA100. In desmutagenic and bioantimutagenic tests, the number of His^＋ revertants of TA98 and TA100 decreased. In the test for spontaneous mutation rate, the number of His＋ revertants of TA98 and TA 100 decreased slightly. When a concentration of CHW was added to examine antimutagenic effects, there was no change in the viable cell count of TA98 and TA100. From these results, AF-2 that is a direct mutagen may combine with CHW, and that may also be carrying out the inactivation. CHW possibly acts directly on the DNA by decreasing the rate at which mutation is caused to the strain＇s promutagens, Trp-P-2 and 2AA. This is evident from the decreased spontaneous mutation rate without a decreased viable cell count.

Antimutagenic and DNA Damage Protective Activities of a Grape Extract from Vitis vinifera

Antimutagenic and DNA protective effect of an extract VinOserae from Vitis vinifera grapes on oxidative DNA damage was investigated. The extract’s ability to inhibit mutagenicity induced by tert-butyl hydroperoxide (t-BHP) and hydrogen peroxide (H2O2) was determined with Ames test using Salmonella typhimurium His? TA102 strain. Inhibition values of 44.2% and 67.0% were detected for t-BHP and H2O2, respectively. A protective ability of the extract against DNA strand scission induced by hydroxyl radicals was studied with plasmid pBluescript II SK(-). The analysis of DNA strand breaks in plasmid DNA showed a significant inhibition of DNA damage.

Organic and Inorganic Pigments from Commercial Inks Used in Eyebrows’ Microblading: Approach for Assessing the Safety Use by Bacterial Reverse Mutation Assay

Background and Objective: The aesthetic products’ safety should not be neglected to the detriment of the market. This study aimed to evaluate the mutagenic potential of commercial inks. It was formulated with organic (P1) and inorganic (P2) pigments for eyebrows microblading using a validated method by regulatory agencies, the Salmonella/microsome assay, to assure the safety of use. Methods: The assay was carried out in three steps: preliminary toxicity, medium without (-S9), and presence (+S9) of metabolic activation. The strains, auxotrophic to histidine (His-) TA97a, TA98, TA100, and TA102, were exposed to both types of inks, in triplicate, compared to negative and positive controls. Data were statistically analyzed, and values with mutagenic index ≥ 2.0 were indicative of mutagenicity. Results: The inks with organic (P1) and inorganic (P2) pigments were not toxic to TA98 and TA100 S. typhimurium tester strains, even at concentrations applied in humans. Both inks were not mutagenic either in the absence or presence of metabolic activation in the tested concentrations, including that applied in humans. The assay showed that P1 and P2 were not direct (-S9) or indirect (+S9) mutagens as commercially formulated. Conclusions: These results indicate that applying these inks on organisms with microsomal enzymes, including humans, is safe since no compound in the inks became more toxic to induce the bacterial reverse mutation.