Genotoxic effects of chlorophenoxy herbicide diclofop-methyl in mice in vivo and in human lymphocytes in vitro

Diclofop-methyl (DM) is a chlorophenoxy derivative used in large quantities for the control of annual grasses in grain and vegetable crops. In this study, the genotoxic effects of DM were investigated by measuring chromosomal aberrations (CAs) in mouse bone-marrow cells and CA and the comet assay in human peripheral lymphocytes. Mice were treated with 15.63, 31.25, 62.5, and 125?mg/kg body weight of DM intraperitoneally for 24 hours, and 15.63-, 31.25-, 62.5-, 125-, and 250-μg/mL concentrations were applied to human lymphocytes for both 24 and 48 hours. In in vivo treatments, DM significantly, but not dose dependently, increased the total chromosome aberrations, compared to both negative and solvent controls. Cell proliferation was significantly, but not dose dependently, affected by all doses. In in vitro treatments, DM (except 15.63 μg/mL) significantly and dose dependently increased the frequency of chromosome aberrations. Also, 250 μg/mL of 48-hour treatment was found to be toxic. Cell proliferation was significantly and dose dependently affected by DM applications, when compared to negative control. In in vitro treatments, DM significantly decreased the mitotic index only at the highest concentration for 24 hours, and 62.5- and 125-μg/mL concentrations for 48 hours. In the comet assay, a significant and dose-dependent increase in comet-tail intensity was observed at 62.5-, 125-, and 250-μg/mL concentrations. The mean comet-tail length was significantly increased in all concentrations. Our results demonstrate that DM is genotoxic in mammalian cells in vivo and in vitro.     

Anti-genotoxic effect of naringin against bleomycin-induced genomic damage in human lymphocytes in vitro

Naringin is a flavonoid found in grapefruit and other citrus fruits that shows antioxidant activity. The aim of the present study was to determine the anti-genotoxic and protective effects of naringin on the chemotherapeutic/radiomimetic agent bleomycin (BLM) in human blood lymphocyte cultures in vitro using micronucleus test and chromosomal aberrations (CA) assay. We tested the three doses of naringin (1, 2, 3?µg/mL) and a single dose of BLM (20?µg/mL). BLM significantly increased the total CAs and micronucleus frequency at a concentration of 20?µg/mL. Naringin did not show any toxicity in doses of 1, 2, and 3?µg/mL. Combined treatments of BLM and naringin (2 and 3?µg/mL) significantly reduced micronucleus formation. Naringin dose-dependently decreased the total chromosome aberrations frequency induced by BLM. These results indicate that naringin could prevent BLM (20?µg/mL)-induced genotoxicity.     

Antimicrobial activity of confertifolin from Polygonum hydropiper

Confertifolin (6,6,9a-trimethyl-4,5,5a,6,7,8,9,9a-octahydronaphtho[1,2-c] furan-3 (1H)-one) was isolated from the essential oil of Polygonum hydropiper L. (Polygonaceae) leaves using column chromatography. Confertifolin showed activity both in bacteria and fungi. The lowest MIC for bacteria was observed against Enterococcus faecalis (31.25 μg/mL). Significant MIC for fungi was observed against Scopulariopsis sp (7.81 μg/mL), Curvularia lunata (7.81 μg/mL), Epidermophyton floccosum (7.81 μg/mL), Trichophyton mentagrophytes (16.62 μg/mL), Trichophyton rubrum (MTCC 296) (16.62 μg/mL), Aspergillus niger (31.25 μg/mL), Botrytis cinerea (31.25 μg/mL) Magnaporthe grisea (62.5 μg/mL), Trichophyton simii (125 μg/mL) and Trichophyton rubrum (clinical isolate) (125 μg/mL).     

Effects of Heliopsis longipes ethanolic extract on mouse spermatozoa in vitro

Context: Heliopsis longipes (A. Gray) Blake (Asteraceae), a plant native to Mexico, is used in traditional medicine as analgesic and microbicide. The main component in the H. longipes ethanolic extract (HLEE) is affinin, as determined by HPLC/UV–visible and NMR measurement. To date, there is no documented evidence on the spermicidal activity of this extract.

Objective: The objective of this study was to assess in vitro the effectiveness of HLEE as spermicide.

Materials and methods: The spermicidal activity of HLEE was evaluated by the Sander–Cramer assay. Spermatozoa were incubated for 20 s with HLEE in concentrations ranging from 75 to 2000 µg/mL to determine the minimum effective concentration (MEC) value. The 50% effective concentration (EC₅₀) of HLEE was estimated by assaying serial dilutions from the MEC. Additionally, sperms were incubated with 125, 250, or 500?µg/mL of HLEE to evaluate the viability and the integrity of sperm membrane. Lipid peroxidation was assessed by the thiobarbituric acid reactive substances assay.

Results: HLEE caused an inhibition of 100% in spermatozoa motility at a MEC value of 2000?µg/mL; the EC₅₀ value was 125?µg/mL. Additionally, exposure to HLEE at 125, 250, or 500?µg/mL for 30?min decreased sperm viability to 27%, 8%, and 2% of the control value, respectively, and significantly increased the percentage of sperms with structurally disorganized membrane. HLEE also increased significantly the level of lipid peroxidation in sperms with respect to controls.

Discussion and conclusion: The results demonstrate the spermicidal activity of HLEE in vitro and suggest that this action is caused by oxidative damage and alterations in the spermatozoal membrane.     

Investigation of genotoxic effects of paraben in cultured human lymphocytes

Paraben is a phenolic derivative of benzoic acid extensively used as preservatives in food, pharmaceutical, and cosmetic industries due to its antimicrobial characteristics. The objective of this study was to evaluate the in vitro genotoxic effects of paraben in human lymphocyte cultures. Cells were analyzed by cytokinesis-block micronucleus (CBMN), chromosome aberration (CA), sister chromatid exchange (SCE), and comet tests. For CBMN, CA, and SCE assays, the human lymphocytes were isolated from healthy donors and incubated with 500, 250, 100, and 50?µg/mL of paraben for 24 and 48?h, and for comet assay, cells were exposed to 1000, 750, 500, and 250?µg/mL of paraben for an hour. Results showed that numbers of MN and SCEs were not significant in the cells exposed to paraben when compared to the solvent control. However, 500 and 250?µg/mL of paraben induced the CA after 24?h. Also, we observed a significant decrease in the cytokinesis-block proliferation index in cells exposed 250–500?µg/mL paraben for 24?h, and 100, 250, and 500?µg/mL for 48?h. The mitotic index was also decreased at all concentrations and periods. However, the proliferation index was statistically decreased at all concentrations after 48?h treatments. Only the highest concentration of paraben caused DNA migration (mean tail length) in human lymphocytes analyzed by Comet assay. Taken together, results indicated that paraben had cytotoxic effects and caused genotoxicity by affecting directly chromosomes and DNA in human lymphocyte cells in vitro, and may have genotoxic potential for human.     

Induction of chromosomal aberrations and micronuclei by 2-hydroxy-4-methoxybenzophenone (oxybenzone) in human lymphocytes

Oxybenzone or benzophenone-3 (2-hydroxy-4-methoxybenzophenone; BP-3) is a filter used in a variety of personal care products for protection of human skin and hair from damage by ultraviolet radiation. BP-3 is suspected to exhibit endocrine disruptive properties. Indeed, it was found to be able to interact with the endocrine system causing alteration of its homeostasis, with consequent adverse health effects. Moreover, it is ubiquitously present in the environment, mostly in aquatic ecosystems, with consequent risks to the health of aquatic organisms and humans. In the present study, we analyzed the cytogenetic effects of BP-3 on human lymphocytes using in vitro chromosomal aberrations and micronuclei assays. Blood samples were obtained from five healthy Italian subjects. Lymphocyte cultures were exposed to five concentrations of BP-3 (0.20, 0.10, 0.05, 0.025, and 0.0125?μg/mL) for 24 and 48?h (for chromosomal aberrations and micronuclei tests, respectively). The concentration of 0.10?µg/mL represents the acceptable/tolerable daily intake reference dose established by European Union, whereas 0.20, 0.05, 0.025, and 0.0125?µg/mL represent multiple and sub-multiple of this concentration value. Our results reported cytogenetic effects of BP-3 on cultured human lymphocytes in terms of increased micronuclei and chromosomal aberrations’ frequencies at all tested concentrations, including concentrations lower than those established by European Union. Vice versa, after 48-h exposure, a significant reduction of the cytokinesis-block proliferation index value in cultures treated with BP-3 was not observed, indicating that BP-3 does not seem to produce effects on the proliferation/mitotic index when its concentration is equal to or less than 0.20?μg/mL.     

Diospyros perigrena bark extract induced apoptosis in filarial parasite Setaria cervi through generation of reactive oxygen species

Abstract

Context: Lymphatic filariasis is a major neglected tropical disease. Diospyros perigrena Gurke (Ebenaceae) was selected for antifilarial chemotherapy because of unavailability of proper medicine. In India, different parts of this plant were used for the treatment of diabetes, diarrhea, dysentery, cholera, mouth ulcers, and wounds.

Objective: The present study was undertaken to access antifilarial potential and mechanism of action of n-butanol extract (NBE) of D. perigrena stem bark on Setaria cervi Rudolphi (Onchocercidae).

Materials and methods: In vitro efficacy and apoptotic mechanism were evaluated by Hoechst, TUNEL, DNA fragmentation assay, pro- and anti-apoptotic gene expression in NBE (250, 125, 62.5, 31.25, and 15.6?µg/ml)-treated S. cervi after 24?h of incubation. Reactive oxygen species (ROS) up-regulation was also determined by GSH, GST, SOD assays, and super oxide anion level.

Results: Significant in vitro antifilarial activity of NBE was found 50% inhibitory concentration (IC₅₀): adult?=?57.6?μg/ml, microfilariae (mf)?=?56.1?μg/ml, and lethal dose (LD₁₀₀) in mf is 187.17?μg/ml) after 24?h of treatment. NBF-induced apoptosis was proved by Hoechst, TUNEL, RT-PCR, and Western blot method. NBF (250?µg/ml) decreased the level of GSH (17.8%) and GST (65.4%), increased SOD activity (1.42-fold) and super oxide anion production (1.32-fold) in the treated parasite which culminated into ROS up-regulation.

Discussion and conclusion: NBE induced apoptosis in different life cycle stages of S. cervi. In future, a detailed study of NBF will give us a novel antifilarial compound which will be used for antifilarial chemotherapy.     

Antioxidant,antibacterial, and anti-inflammatory activities of standardized brazilin-rich Caesalpinia sappan extract

Abstract

Context: Brazilin is a major active principle of Caesalpinia sappan L. (Leguminosae or Fabaceae). For industry aspects, brazilin-rich extract (BRE) has been prepared and standardized to contain 39% w/w brazilin. BRE may have more advantages than brazilin in term of a lower-cost production process.

Objectives: To investigate the antioxidant, antibacterial, and anti-inflammatory activities of BRE.

Material and methods: BRE was prepared by a simple one-step purification of the crude ethanol extract of C. sappan heartwood (CSE) using a Diaion® HP-20 column. The antioxidant activities were determined using three methods, including DPPH radical scavenging, reducing power, and β-carotene bleaching assays, at concentration ranges of 1–10, 10–100, and 10–100?µg/mL, respectively. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of BRE (15.6–1000?µg/mL) against Gram-positive and Gram-negative bacteria were determined by the broth microdilution method. Anti-inflammatory activity of BRE (0.1–5?µg/mL) was evaluated as anti-denaturation activity using bovine serum albumin as a substrate.

Results and discussion: On the basis of β-carotene bleaching assay, BRE showed antioxidant activity with an EC₅₀ value of 60.5?µg/mL, which was almost equal to that of pure brazilin (52.1?µg/mL). Gram-positive bacteria were more sensitive to all tested samples than Gram-negative bacteria. BRE possessed higher antibacterial activities than CSE, but lower than brazilin. MIC/MBC values of 62.5–125/125 and 250–500/250–500?µg/mL were obtained for BRE against Gram-positive and Gram-negative bacteria, respectively. A low concentration (0.1?µg/mL) of brazilin, BRE, and CSE showed anti-inflammatory activity by inhibiting protein denaturation up to 46.8, 54.1, and 61.9%, respectively.     

Perfluorooctanoic acid‐induced toxicity in primary cultures of chicken embryo cardiomyocytes

Perfluorooctanoic acid (PFOA) is a widespread environmental contaminant that induces developmental cardiotoxicity. It is detectable in late stage chicken embryos and hatchling chickens. To investigate mechanism(s) of cardiotoxicity, primary cultures of cardiomyocytes were prepared from 10‐day‐old chicken embryos that were (A) pre‐exposed to vehicle or 2 mg of PFOA/kg of egg weight in ovo or (B) incubated with PFOA in vitro at concentrations ranging from 0 to 100 µg/mL in medium for 1 or 36 h. When viability was assessed, survival of cardiomyocytes prepared from pre‐exposed embryos did not differ from vehicle controls, even under conditions of serum starvation designed to challenge the cells. However, 1 h of exposure to 100 µg/mL of PFOA in vitro and 36 h of exposure to 75 and 100 µg/mL PFOA in vitro decreased viability. When contractility was evaluated, cardiomyocytes cultured from pre‐exposed embryos exhibited decreases in time to maximum departure velocity and cell length at peak contraction, whereas cardiomyocytes exposed in vitro exhibited a reduction in the 50% relaxation time at a concentration of 1 µg/mL relative to vehicle controls. Morphological assessment revealed decreased cardiomyocytes axial length following in ovo PFOA exposure and 24 h in vitro PFOA 50 µg/mL exposure. Reactive oxygen species (ROS) generation, which was evaluated only in cardiomyocytes exposed to PFOA in vitro, was significantly elevated following incubation with 50 µg/mL of PFOA for 1 h. These data indicate that while in vitro exposure to relatively high concentrations of PFOA can induce cytotoxicity and ROS, developmental cardiotoxicity observed in ovo is not likely mediated via PFOA‐induced overt cytotoxicity, but likely by altering early cardiac morphologic and function processes. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1580–1590, 2016.     

Chromosomal aberrations in cultured human lymphocytes treated with the fungicide,Thiram

In vitro effects of different concentrations of Thiram were tested on human lymphocytes to determine, by means of the chromosome aberrations (CAs) assay, whether this fungicide could induce clastogenic damage. Evidences of the effect of Thiram on human lymphocytes were limited to sister chromatid exchange, micronuclei formation, and comet assays. We evaluated 0.01, 0.1, 1.2, and 12.0 μg/mL of Thiram, where 0.01 μg/mL represent the acceptable daily intake dose set by the World Health Organization and the Food and Agriculture Organization for fruit and vegetables, whereas 0.1, 1.2, and 12.0 μg/mL are its multiple values. Results indicated that human lymphocytes treated in vitro with Thiram at concentrations of 1.20 and 12.0 μg/mL significantly increased CAs frequency, compared with the negative control, whereas at lower concentrations (0.01 and 0.1 μg/mL), this effect was not observed. However, Thiram showed a clastogenic effect also at the concentration value of 1.2 μg/mL that represents a lower value with respect to the residue limits found in Italy for grapes, strawberries, potatoes, tobacco, and other fruits and vegetables. Finally, according to some evidence obtained from the study of other fungicides, Thiram produced a significant reduction in the mitotic index with increasing concentration.     

Genotoxicity of monosodium glutamate

Monosodium glutamate (MSG) is one of the most widely used flavor enhancers throughout the world. The aim of this study is to investigate the genotoxic potential of MSG by using chromosome aberrations (CAs), sister-chromatid exchanges (SCEs), cytokinesis-blocked micronucleus (CBMN), and random amplified polymorphic DNA-polimerase chain reaction (RAPD-PCR) in cultured human lymphocytes and alkaline comet assays in isolated human lymphocytes, which were incubated with six concentrations (250, 500, 1000, 2000, 4000 and 8000 μg/mL) of MSG. The result of this study indicated that MSG significantly and dose dependently increased the frequencies of CAs, SCE and MN in all treatments and times, compared with control. However, the replication (RI) and nuclear division indices (NDI) were not affected. In this paper, in vitro genotoxic effects of the MSG was also investigated on human peripheral lymphocytes by analysing the RAPD-PCR with arbitrary 10-mer primers. The changes occurring in RAPD profiles after MSG treatment include increase or decrease in band intensity and gain or loss of bands. In the comet assay, this additive caused DNA damage at all concentrations in isolated human lymphocytes after 1-h in vitro exposure. Our results demonstrate that MSG is genotoxic to the human peripheral blood lymphocytes in vitro.     

Determination of the toxicity of intravitreal minocycline in rabbit eyes

Purpose: To evaluate the retinal toxicity of intravitreal minocycline in rabbit eyes.

Methods: Intravitreal injection of minocycline with concentrations of 1000, 500, 250, 125 and 62.5?μg in 0.1?ml was performed in 10 New Zealand albino rabbits. Each concentration was injected into two rabbit eyes. For each dose, normal saline was injected in one contralateral eye and the other fellow eye remained non-injected. Electrophysiologic testing was performed before and 4 weeks after injections. The eyes were enucleated 4 weeks after injections and examined using light microscopy.

Results: The clinical examination was unremarkable after injections. Electroretinography recordings were significantly affected at all doses in at least one of the a- or b-waves of photopic or scotopic responses. Histopathologic examination revealed marked atrophy and loss of integrity in all retinal layers in all minocycline injected eyes. Contralateral eyes were normal.

Conclusion: In our study, intravitreal minocycline was toxic to the retina in albino rabbits even at a concentration of 62.5?µg/0.1?ml.     

Pheretima aspergillum extract attenuates high‐KCl‐induced mitochondrial injury and pro‐fibrotic events in cardiomyoblast cells

Hyperkalemia is often associated with cardiac dysfunction. In this study an earthworm extract (dilong) was prepared from dried Pheretima aspergillum powder and its effect against high‐KCl challenge was determined in H9c2 cardiomyoblast cells. H9c2 cells pre‐treated with dilong (31.25, 62.5, 125, and 250 mg/mL) for 24 hours, where challenged with different doses of KCl treatment for 3 hours to determine the protective mechanisms of dilong against cardiac fibrosis. High‐KCl administration induced mitochondrial injury and elevated the levels of pro‐apoptotic proteins. The mediators of fibrosis such as ERK, uPA, SP1, and CTGF were also found to be upregulated in high‐KCl condition. However, dilong treatment enhanced IGF1R/PI3k/Akt activation which is associated with cell survival. In addition, dilong also reversed high‐KCl induced cardiac fibrosis related events in H9c2 cells and displayed a strong cardio‐protective effect. Therefore, dilong is a potential agent to overcome cardiac events associated with high‐KCl toxicity.     

Adverse Male Reproductive Effects following Subchronic Exposure of Rats to Sodium Dichloroacetate

Dichloroacetate (DCA) activates the pyruvate dehydrogenase complexenhancing carbohydrate and lactate utilization in animals. Asa result it is used clinically in the treatment of acute lacticacidosis and has therapeutic potential in the treatment of stroke.Adverse effects of chronic DCA treatment include poly-neuropathyand testicular degeneration. Since DCA is a principal productof the aqueous chlorination of fulvic acids concern has arisenregarding the agent's impact on environmental health. We treatedmale Long-Evans rats with 0, 31.25, 62.5, or 125 mg DCA/kg/dayby oral gavage for 10 weeks. Compared to controls, preputialgland and epididymis weights were reduced at 31.25 mg/kg, bodyand liver weights at 62.5 mg/kg, and accessory organ weightsat 125 mg/kg. Epididymal sperm counts were reduced and spermmorphology was impacted at the 62.5 and 125 mg/kg doses levels.Histologic examination of the testis and epididymis revealedinhibited spermiation in testes at the 125 mg/kg dose level.Computer-assisted sperm motion analysis revealed reductionsin percentage motile sperm, curvilinear and straight-line velocity,linearity, and amplitude of lateral head displacement at boththe 62.5 and the 125 mg/kg dose levels. In the assessment offertility after an overnight mating, the number of viable implantson Day 14 of gestation was decreased only in the highest dosegroup. These studies demonstrate adverse effects of NaDCA treatmenton the rat male reproductive system, primarily on the accessoryorgans and sperm within them at lower doses (31.25 and 62.5mg/kg), and on the testis at the highest dOSe (125 mg/kg).     

Bioassay-Guided Isolation of Antimalarial Triterpenoid Acids from the Leaves of Morinda lucida.

Abstract

The EtOH, CH₂Cl₂, and petroleum ether extracts from Morinda lucida. Benth. leaves have been shown to exhibit an in vitro. antiplasmodial activity against a chloroquine-sensitive Plasmodium falciparum. strain with IC₅₀ values 5.7 ± 1.3, 5.2 ± 0.8, and 3.9 ± 0.3 µg/mL, respectively. In vivo., at a daily oral dose of 200 mg/kg body weight, they produced at least 62.5%, 67.5%, and 72.2% reduction of parasitemia in mice infected with Plasmodium berghei berghei., respectively. A bioassay-guided fraction of the most active petroleum ether extract resulted in the isolation of two known triterpenic acids as ursolic acid 1 and oleanolic acid 2. In vitro., 1 and 2 exhibited an antiplasmodial activity with IC₅₀ values of 3.1 ± 1.3 and 15.2 ± 3.4 µg/mL, respectively. In vivo., at a daily dose of 200 mg/kg body weight, they produced 97.7% and 37.4% chemosuppression, respectively. However, all tested samples were inactive in vitro. against chloroquine-resistant Plasmodium falciparum. (K1) at the highest tested concentration of 25 µg/mL.     

Evaluation of cytogenetic and DNA damage caused by thallium(I) acetate in human blood cells

Although thallium is detrimental to all living organisms, information regarding the mutagenic and genotoxic effects of this element and its compounds remains scarce. Therefore, we tested the genotoxic and cytotoxic effects of thallium(I) acetate on human peripheral blood cells in vitro using structural chromosomal aberrations (SCAs), sister chromatid exchanges (SCEs), and single‐cell gel electrophoresis (at pH >13 or 12.1) analysis. Whole blood samples were incubated with 0.5, 1, 5, 10, 50, or 100 µg/mL thallium salt. Exposure to this metal compound resulted in a clear dose‐dependent reduction in the mitotic and replicative indices. An increase in SCAs was evident in the treated group compared with the control group, and significant differences were observed in the percentage of cells with SCAs when metaphase cells were treated with 0.5–10 µg/mL of thallium(I). The SCE test did not reveal any significant differences. We observed that a 1‐h treatment with thallium(I) at pH > 13 significantly increased the comet length for all the concentrations tested; however, at pH 12.1, only the two highest concentrations affected the comet length. These results suggested that thallium(I) acetate induces cytotoxic, cytostatic, and clastogenic effects, as well as DNA damage. © 2013 Wiley Periodicals, Inc. Environ Toxicol 30: 572–580, 2015.     

Evaluation of cytogenetic and DNA damage induced by the antidepressant drug-active ingredients,trazodone and milnacipran,in vitro

Trazodone and milnacipran are the active antidepressant drugs that are being used in the treatment of psychiatric disorders. In this study, the in vitro genotoxic effects of trazodone and milnacipran have been determined in human peripheral blood lymphocytes by using chromosomal aberrations (CAs), sister chromatid exchanges (SCEs), micronuclei (MN), and comet assays. 3.13; 6.25; 12.50; 25.00; 50.00; and 75.00?μg/mL concentrations of trazodone and 2.50; 5.00; 10.00; 20.00; 30.00; and 40.00?μg/mL concentrations of milnacipran were used. Trazodone and milnacipran significantly increased the frequency of CAs and SCEs compared with the control. Both of the active ingredients raised the MN frequency in a dose-dependent manner. Mitotic index was significantly decreased, but replication and nuclear division indices were not affected at all treatments. Trazodone was statistically increased the mean comet tail intensity, tail length, and tail moment at three concentrations (6.25; 12.50; and 25.00?μg/mL) compared with control. Two highest concentrations (50 and 75?μg/mL) of trazodone were toxic in the comet assay. Milnacipran increased the comet tail intensity, tail length, and tail moment at all concentrations. It is concluded that trazodone and milnacipran have clastogenic, mutagenic, and cytotoxic effects on human lymphocytes in vitro.     

Chemical constituents and biological activities of Albizia myriophylla wood

Context: Albizia myriophylla Benth (Leguminosae) is a medicinal plant widely used in Thailand and other Asian countries as a folk medicine remedy for many ailments.

Objective: The objective of this study is to investigate the chemical compositions, antibacterial activity, and cytotoxicity of A. myriophylla wood.

Materials and methods: The structure identification of the isolated compounds was established using spectroscopic methods. In vitro antibacterial activity against Streptococcus mutans, Staphylococcus aureus, and Bacillus cereus and the cytotoxicity against KB cells of extracts and compounds from A. myriophylla were performed using broth microdilution and resazurin microplate assays, respectively. The lupinifolin content in A. myriophylla extracts was quantified by high-performance liquid chromatography.

Results: A rare flavan-3,4-diol (1) together with eight known compounds (2–9) were isolated from the wood of A. myriophylla. Compounds 4–9 exhibited anti-S. mutans activity, of which lupinifolin (5) was the most potent with an MIC value of 0.98?µg/mL, followed by its dihydroxy derivative 4 with an MIC value of 62.5?µg/mL. Compounds 4 and 5 also displayed marked antibacterial activity against B. cereus and S. aureus (MIC value 15.63–125?µg/mL) and showed strong cytotoxic activity against KB cells (IC₅₀ value 4.95–12.55?µg/mL). The lupinifolin contents in ethanol extracts from two different collections of this plant originating from central and southern Thailand were 93.85 and 0.04?mg/g, respectively.

Conclusion: This is the first report of compounds 1–4 from A. myriophylla. Compounds 4 and 5 showed potent antibacterial and cytotoxic activities compared with other isolates. The anti-S. mutans activity of A. myriophylla extracts seems to be related to the lupinifolin content.     

In vitro cytotoxicity and genotoxicity of Furia®180 SC (zeta-cypermethrin) and Bulldock 125®SC (β-cyfluthrin) pyrethroid insecticides in human peripheral blood lymphocytes

In the present study, human peripheral blood lymphocytes were exposed in vitro to 0, 6, 12, 18, 24, and 30?μg/mL Furia^®180 SC (zeta-cypermethrin) and 0, 6.3, 12.5, 18.8, 25, and 31.3?μg/mL Bulldock^®125 SC (β-cyfluthrin). Exposure to 32?µg/mL bleomycin for 24?h served as a positive control. The cytotoxic and genotoxic effects of each insecticide were analyzed using alkaline comet and trypan blue dye exclusion assays. DNA damage was evaluated through three genotoxicity parameters: tail length (TL), tail moment (TM) and tail intensity (TI). Furia^®180 SC and Bulldock^®125 SC pyrethroid insecticides and bleomycin significantly increased DNA damage in a concentration-dependent manner. Bulldock^®125 SC induced more DNA damage than Furia. Lymphocyte viability did not change after exposure to different concentrations of the two pyrethroid insecticides and bleomycin. Moreover, genotoxic results demonstrated that Furia^®180 SC and Bulldock^®125 SC insecticides caused in vitro DNA damage in human peripheral lymphocytes.     

Genotoxicity of Aspartame

In the present study, the genotoxic effects of the low‐calorie sweetener aspartame (ASP), which is a dipeptide derivative, was investigated using chromosome aberration (CA) test, sister chromatid exchange (SCE) test, micronucleus test in human lymphocytes and also Ames/Salmonella/ microsome test. ASP induced CAs at all concentrations (500, 1000 and 2000 µg/ml) and treatment periods (24 and 48 h) dose‐dependently, while it did not induce SCEs. On the other hand, ASP decreased the replication index (RI) only at the highest concentration for 48 h treatment period. However, ASP decreased the mitotic index (MI) at all concentrations and treatment periods dose‐dependently. In addition, ASP induced micronuclei at the highest concentrations only. This induction was also dose‐dependent for 48 hours treatment period. ASP was not mutagenic for Salmonella typhimurium TA98 and TA100 strains in the absence and presence of S9 mix.