Cytotoxic and genotoxic effects of <Emphasis Type="Italic">Lavandula stoechas</Emphasis> aqueous extracts

Lavandula genus is an important member of Labiatae (Lamiaceae) family. People use commonly Lavandula stoechas as a medicinal plant for various diseases around the world and also in Turkey. The aim of this study was to investigate cytotoxic and genotoxic effects of aqueous extracts (40, 80 and 120 g/L) from L. stoechas flowers on Allium cepa root tip meristem cells. For this purpose, A. cepa onion bulbs were treated with the above-mentioned L. stoechas flower extracts for 72 h. Spring water (pH 7.3) was used as a control. The result of this study sowed that aqueous extracts reduced mitotic index, but induced chromosome aberrations and mitotic aberrations in comparison with control, significantly (p < 0.05). Aqueous extracts induced breaks, stickiness, pole deviations and micronuclei. Furthermore, these effects were related to extract concentrations. These results showed that L. stoechas aqueous extracts have cytotoxic and genotoxic effects.     

Cytogenetic effects of silver and gold nanoparticles on Allium cepa roots

The present study evaluates the cytogenetic effects of both silver and gold nanoparticles on the root cells of Allium cepa. In this study, the root cells of Allium cepa were treated with both gold and silver nanoparticles of different concentrations (1?mg/L, 5?mg/L and 10?mg/L) along with control for 72?h. Experimental results revealed that after 72?h of exposure, a significant decrease in mitotic index (MI) from 68% (control) to 52.4% (1?mg/L), 47.3% (5?mg/L) and 41.4% (10?mg/L) for gold nanoparticles and 57.1% (1?mg/L), 53% (5?mg/l), 55.8% (10?mg/L) for silver nanoparticles. Through minute observation of the photograph, it was recorded that some specific chromosomal abnormalities such as stickiness of chromosome, chromosome breaks, nuclear notch, and clumped chromosome at different exposure conditions. Therefore, present results clearly suggest that Allium cepa root tip assay could be a viable path through which negative impact of both gold and silver nanoparticles can be demonstrated over a wide range of concentrations.     

Genotoxicity of five food preservatives tested on root tips of Allium cepa L.

The effects of the food preservatives sodium benzoate (SB), boric acid (BA), citric acid (CA), potassium citrate (PC) and sodium citrate (SC) have been studied on root tips of Allium cepa L. Roots of A. cepa were treated with a series of concentrations, ranging from 20 to 100 ppm for 5, 10 and 20 h. The results indicate that these food preservatives reduced mitotic division in A. cepa compared with the respective control. Mitotic index values were generally decreased with increasing concentrations and longer treatment times. Additionally, variations in the percentage of mitotic stages were observed. The total percentage of aberrations generally increased with increasing concentrations of these chemicals and the longer period of treatment. Different abnormal mitotic figures were observed in all mitotic phases. Among these abnormalities were anaphase bridges, C-mitosis, micronuclei, lagging, stickiness, breaks and unequal distiribution.     

Genotoxicity of drinking water disinfection by-products (bromoform and chloroform) by using both Allium anaphase-telophase and comet tests

Genotoxic effects of bromoform and chloroform, disinfection by-products of the chlorination of drinking water, were examined by using mitotic index (MI), mitotic phase, chromosome aberrations (CAs) and comet assay on root meristematic cells of Allium cepa. Different concentrations of bromoform (25, 50, 75 and 100 μg/mL) and chloroform (25, 50, 100 and 200 μg/mL) were introduced to onion tuber roots. Distilled water was used as a negative control and methyl methansulfonate (MMS-10 μg/mL) as positive control. All obtained data were subjected to statistical analyses by using SPSS 15.0 for Windows software. For comparison purposes, Duncan multiple range tests by using one-way analysis of variance were employed and p < 0.05 was accepted as significant value. Exposure of both chemicals (except 25 μg/mL applications of bromoform) significantly decreased MI. Bromoform and chloroform (except 25 μg/mL applications) increased total CAs in Allium anaphase-telophase test. A significant increase in DNA damage was also observed at all concentrations of both bromoform and chloroform examined by comet assay. The damages were higher than that of positive control especially at 75–100 μg/mL for bromoform and 100–200 μg/mL for chloroform.     

Cytogenotoxic effects of ethanolic extracts of Annona crassiflora (Annonaceae)

Infusions of the leaves and seeds of Annona crassiflora Mart. are commonly employed in the treatment of diarrhoea, snakebites, tumours and disorders of the hair and scalp. The aim of the present study was to investigate the cytotoxic and genotoxic properties of ethanolic extracts of A. crassiflora by evaluating their effects on germination, root elongation, chromosome structure and the cell division of Lactuca sativa (lettuce). The experiment followed a randomized design involving the treatment of L. sativa seeds with ethanolic extracts from leaves and seeds of A. crassiflora applied at ten concentrations (0.0125, 0.025, 0.05, 0.1, 0.2, 0.4, 0.6, 0.8, 1.0, and 1.2 mg/L) and with five repetitions per treatment. Seeds of L. sativa exposed for 48 h to A. crassiflora leaf extract at concentrations ≥ 0.1 mg/L, or to seed extracts at concentrations ≥ 0.2 mg/L, showed germination percentages that were significantly (p < 0.05) lower than those of seeds exposed to aqueous ethanol control. Exposure of L. sativa seedlings to leaf (but not seed) extracts of A. crassiflora produced significant (p < 0.05) reductions in the mitotic indices of root meristem cells of lettuce and induced chromosome and nuclear abnormalities in the root cells. The presence of chromosome stickiness, bridges, fragments, laggard chromosomes and nuclear condensation were also observed. The cytogenetic effects observed suggest that folkloric medicines prepared with extracts of the leaves or seeds of A. crassiflora should be employed with caution.     

Comparison of cytotoxic and genotoxic effects of plutonium-239 alpha particles and mobile phone GSM 900 radiation in the Allium cepa test

The goal of this study was to compare the cytotoxic and genotoxic effects of plutonium-239 alpha particles and GSM 900 modulated mobile phone (model Sony Ericsson K550i) radiation in the Allium cepa test. Three groups of bulbs were exposed to mobile phone radiation during 0 (sham), 3 and 9 h. A positive control group was treated during 20 min with plutonium-239 alpha-radiation. Mitotic abnormalities, chromosome aberrations, micronuclei and mitotic index were analyzed. Exposure to alpha-radiation from plutonium-239 and exposure to modulated radiation from mobile phone during 3 and 9 h significantly increased the mitotic index. GSM 900 mobile phone radiation as well as alpha-radiation from plutonium-239 induced both clastogenic and aneugenic effects. However, the aneugenic activity of mobile phone radiation was more pronounced. After 9 h of exposure to mobile phone radiation, polyploid cells, three-groups metaphases, amitoses and some unspecified abnormalities were detected, which were not registered in the other experimental groups. Importantly, GSM 900 mobile phone radiation increased the mitotic index, the frequency of mitotic and chromosome abnormalities, and the micronucleus frequency in a time-dependent manner. Due to its sensitivity, the A. cepa test can be recommended as a useful cytogenetic assay to assess cytotoxic and genotoxic effects of radiofrequency electromagnetic fields.     

The genotoxic and anti-genotoxic effects of <Emphasis Type="Italic">Stachys petrokosmos</Emphasis> leaf extract in human lymphocytes using microsomal fractions

The genotoxic and anti-genotoxic effects of Stachys petrokosmos leaf extracts (Sp) were investigated in human lymphocytes. The cells were treated with 1.5, 3.0 and 6.0 μL/mL concentrations of Sp leaf extracts for 24 and 48 h treatment periods in the absence and presence of metabolic activator (S9mix). In the absence of S9mix, Sp alone did not induce chromosome aberrations and formation of micronucleus while inducing the mean sister chromatid exchange at the highest concentration. In addition, Sp decreased the mutagenic effect of mitomycin-c. Sp alone showed a cytotoxic effect determined by a decrease in the proliferation index, mitotic index and nuclear division index. On the other hand a mixture of Sp and mitomycin-c resulted in a higher cytotoxic effect especially for 48 h treatment period. In the presence of S9mix, Sp was not genotoxic and cytotoxic however, it showed an anti-genotoxic effect by decreasing the effects of cyclophosphamide.     

Cytogenetic effect of prolonged in vitro exposure of <Emphasis Type="Italic">Allium cepa</Emphasis> L. root meristem cells to salt stress

Salt stress severely affects plant growth and regulation; however knowledge about its effect on cytological changes is limited. In this report, studies were carried out to reveal the morphological and cytogenetic responses of the Allium cepa roots after prolonged (7, 14, 20 days) exposure to salt stress in tissue culture conditions. Roots of plants were treated with different concentrations (100, 200, 300 mM) of NaCl and KCl added to the culture medium. Both salts caused root growth reduction and showed cytotoxic effect reflected in reduction of root tip cells mitotic activity and increase of different abnormalities such as chromosome breaks and bridges and formation of micronuclei. This experiment showed differences of the action of excessive amounts of both tested salts at the cellular level, as KCl induced higher frequencies of abnormalities during cell divisions, whereas NaCl showed more mitodepressive effect and more frequently led to the root meristem cell death.     

Cytotoxic effects of DON and T-2 toxin on plant cells

The effect of deoxynivalenol (DON) and T-2 toxin on mitotic index (MI) and relative division rate (RDR) in actively dividing onion (Allium cepa L.) root-tip cells was studied. Both these toxins resulted in decline of mitotic activity which was inversely proportional to the concentrations of these toxins. T-2 was more effective resulting in 59% RDR value at 2.5 ppm whereas DON treated root cells had 78% RDR at the same concentration as compared to respective sets of controls.     

Anti-mitotic and anti-genotoxic effects of Plantago lanceolata aqueous extract on Allium cepa root tip meristem cells

Plantago is the most important genus of Plantaginaceae family and is used in traditional medicine around the world for different purposes. Plantago coronopus L., Plantago major L., Plantago media L. and Plantago lanceolata L. are most commonly used species of Plantago in traditional medicine in Turkey. The main goal of this study was to investigate the eventual anti-mitotic and anti-genotoxic effects of P. lanceolata L. leaf aqueous extracts (15 g/L and 30 g/L) on Allium cepa L. root tip meristem cells which were treated with 0.7% hydrogen peroxide. For this purpose, two different experiments were performed under the same conditions. In the first experiment, Allium cepa onion bulbs were treated with 0.7% H₂O₂ for 1 h. After the H₂O₂ treatment, the onion bulbs were treated with two different concentrations (15 g/L and 30 g/L) of P. lanceolata extracts for 24 h. In the second experiment, A. cepa onion bulbs were treated with two different extract concentrations (15 g/L and 30 g/L) for 24 h and then with 0.7% H₂O₂ for 1 h. The test concentrations were determined according to doses which are recommended in alternative medicinal usage by people. As positive and negative control 0.7% H₂O₂ and tap water was used, respectively. As a result, it was determined that aqueous extracts reduced mitotic index and chromosome aberrations in treatment groups in comparison with controls. These results showed that P. lanceolata aqueous extracts have anti-mitotic and anti-genotoxic effects.     

Chromosome Aberrations and Ageing Root Meristems

Changes in mitotic activity and frequency of spontaneous chromosomeaberrations have been determined for primary roots of Zea muys,Pisum sativum, Vicia faba var. green Windsor and var. minor,and Allium cepa. Mitotic activity was found to decline withage in all root apices. In the primary root apices immediately after germination, therewas a high frequency of spontaneous chromosome aberrations whichdeclined with time after germination. However, In the case ofV. faba var. green Windsor, Z mays and P. sativum there wasa return to the high levels of spontaneous chromosome aberrationon ageing of the root apex. root meristems, mitotic activity, chromosome aberrations, ageing     

Phenolic acid allelochemicals induced morphological,ultrastructural, and cytological modification on <Emphasis Type="Italic">Cassia sophera</Emphasis> L. and <Emphasis Type="Italic">Allium cepa</Emphasis> L.

The allelopathic potential of leaf aqueous extract (LAE) of Calotropis procera on growth behavior, ultrastructural changes on Cassia sophera L., and cytological changes on Allium cepa L. was investigated. LAE at different concentrations (0.5, 1, 2, and 4 %) significantly reduced the root length, shoot length, and dry biomass of C. sophera. Besides, the ultrastructural changes (through scanning electron microscopy, SEM) induced in epidermal cells of 15-day-old seedlings of Cassia leaf were also noticed. The changes induced were shrinking and contraction of epidermal cells along with the formation of major grooves, canals, and cyst-like structures. The treated samples of epidermal cells no longer seem to be smooth as compared to control. LAE at different concentrations induces chromosomal aberrations and variation in shape of the interphase and prophase nucleus in A. cepa root tip cells when compared with control groups. The mitotic index in treated onion root tips decreased with increasing concentrations of the extracts. The most frequent aberrations were despiralization at prophase with the formation of micronuclei, sticky anaphase with bridges, sticky telophase, C-metaphase, etc. The results also show the induction of ghost cells, cells with membrane damage, and cells with heterochromatic nuclei by extract treatment. Upon HPLC analysis, nine phenolic acids (caffeic acid, gentisic acid, catechol, gallic acid, syringic acid, ellagic acid, resorcinol, p-coumaric acid, and p-hydroxy benzoic acid) were identified. Thus, the phenolic acids are mainly responsible for the allelopathic behavior of C. procera.     

Allium cepa assay based comparative study of selected vegetables and the chromosomal aberrations due to heavy metal accumulation

Irrigation of industrial effluents may end in the bioaccumulation of various toxic metals and consequent genetic changes in contaminated food crops. To test this hypothesis and extent of genetic modifications, Allium cepa test was performed to food crops viz. tomato (Lycopersicum esculentum) and chili (Capsicum annum) as Allium cepa test is a useful tool to assess genetic variations in plants. Prior to A. cepa test, the plants were exposed to various metal concentrations 125–1000 mg/L in the synthetic wastewater. The extracts of harvested plants were used to grow the root of A. cepa following its standard method. The root tips were fixed, stained and examined under compound microscope (almost 300–400 dividing cells) to check the extent of chromosomal variations during various stages of mitosis. The results revealed various chromosomal abnormalities including laggards, stickiness, vagrant chromosomes, binucleated cells, nuclear lesions, giant cells and c-mitosis at different level of treatment. On the whole, aberrations were increasing with the increasing doses along the positive control. In comparison, chili crop had higher level of aberrations depicting the higher chromosomal changes. Lower mitotic index (MI) with increasing level of doses was also describing the hampered cell division due to increased metal stress. The study is showing that the cell division was ceased with increasing metal stress thus increasing the rate of cell aberrations.     

Genotoxicity of Thermopsis turcica on Allium cepa L. roots revealed by alkaline comet and random amplified polymorphic DNA assays

This study was undertaken to evaluate genotoxic potential of Thermopsis turcica aqueous extracts on the roots of onion bulb (Allium cepa L.) by comet assay and random amplified polymorphic DNA technique. The Allium root growth inhibition test indicated that the EC₅₀ and 2×EC₅₀ values were 8 and 16 mg/ml concentrations of T. turcica aqueous extracts, respectively. The negative control (distilled water), positive control (methyl methane sulfonate, 10 mg/l) and 8 and 16 mg/ml concentrations of T. turcica extracts were introduced to the roots of onion bulbs for 24 and 96 h. The root growth, DNA damage in root cells and randomly amplified polymorphic DNA (RAPD) profiles of root tissue were used as endpoints of the genotoxicity. The comet assay clearly indicated that dose-dependent single strand DNA breaks in the root nuclei of onions were determined for the treatment concentrations of T. turcica extracts. In comparison to RAPD profile of negative control group, RAPD polymorphisms became evident as disappearance and/or appearance of RAPD bands in treated roots. The diagnostic and phenetic numerical analyses of RAPD profiles obviously indicated dose-dependent genotoxicity induced by Thermopsis extracts. In conclusion, the results clearly indicated that water extract of T. turcica has genotoxic potential on the roots of onion bulbs as shown by comet assay and RAPD technique.     

Genotoxicity assessment of a pharmaceutical effluent using four bioassays

Pharmaceutical industries are among the major contributors to industrial waste. Their effluents when wrongly handled and disposed of endanger both human and environmental health. In this study, we investigated the potential genotoxicity of a pharmaceutical effluent, by using the Allium cepa, mouse- sperm morphology, bone marrow chromosome aberration (CA) and micronucleus (MN) assays. Some of the physico-chemical properties of the effluent were also determined. The A. cepa and the animal assays were respectively carried out at concentrations of 0.5, 1, 2.5, 5 and 10%; and 1, 5, 10, 25 and 50% of the effluent. There was a statistically different (p < 0.05), concentration-dependent inhibition of onion root growth and mitotic index, and induction of chromosomal aberrations in the onion and mouse CA test. Assessment of sperm shape showed that the fraction of the sperm that was abnormal in shape was significantly (p < 0.05) greater than the negative control value. MN analysis showed a dose-dependent induction of micronucleated polychromatic erythrocytes across the treatment groups. These observations were provoked by the toxic and genotoxic constituents present in test samples. The tested pharmaceutical effluent is a potentially genotoxic agent and germ cell mutagen, and may induce adverse health effects in exposed individuals.     

Allelopathic potential of two sesquiterpene lactones from Magnolia grandiflora L.

The allelopathic effects of the two sesquiterpene lactones, costunolide and parthenolide, isolated from the leaves of Magnolia grandiflora L. were evaluated on the wheat (Triticum aestivum L.), lettuce (Lactuca sativa L.), radish (Raphanus sativus L.) and onion (Allium cepa L.). Seed germination of the test species was significantly reduced at 500 μg/ml by both compounds. Both sesquiterpenes showed pronounced inhibition of root length of the test species and the inhibitory effect was concentration-dependent. In addition, shoot growth of the four species was significantly inhibited at all the concentrations tested (10–500 μg/ml). Parthenolide reduced germination and inhibited seedling growth more than costunolide. Inhibition of root growth was generally greater than that of shoot growth. The results encourage the use of these sesquiterpenes as models for development of new herbicides.     

Extracts affecting mitotic division in root-tip meristematic cells

Some species of Psychotria (Rubiaceae) are important in herbal medicine, where their extracts are used internally for infections of the female reproductive system, bronchitis, gastrointestinal disturbances, skin irritations, tumors, ulcers, and eye disturbances. The antiproliferative and genotoxic effects of Psychotria myriantha Mull. Arg. and P. leiocarpa Cham. et Schlecht infusions on the Allium cepa L. cell cycle were evaluated. The teas were prepared by infusing the leaves in distilled water, in two concentrations: 1.13 mg/mL and 6.78 mg/mL. Three groups of four bulbs were used for each Psychotria species. After the bulbs rooted in distilled water, they were transferred to the teas for 24 hours, except for the control that stayed in water. The rootlets were then collected, fixed in ethanol-acetic acid (3:1) for 24 hours, and stored in 70% ethanol. For each group of bulbs, 4000 cells were analyzed, calculating the mitotic indexes, submitting them to statistic analysis, using the χ ² test (p = 0.05). The results showed a decrease in mitotic index with an increase in tea concentration in both species. In P. leiocarpa, the mitotic index values differed significantly between the control and concentration of 6.78 mg/mL (χ ² = 9.863). For P. myriantha, the values referring to the mitotic index differed greatly between the control and the treatments (χ ² = 124.8). With this study, it was determined that P. myriantha and P. leiocarpa infusions possess antiproliferative effects on the A. cepa cell cycle, and teas of P. myriantha also have genotoxic activity.     

The effects of 4-thujanol on chromosome aberrations,sister chromatid exchanges and micronucleus in human peripheral blood lymphocytes

4-Thujanol, a bicyclic monoterpene alcohol, is present in the essential oils of many medicinal and aromatic plants. It is commonly used as a fragrance and flavouring ingredient in a lot of different products. The potential genotoxic effects of 4-thujanol on human peripheral blood lymphocytes (PBLs) were investigated in vitro by the chromosome aberrations (CAs), sister chromatid exchanges (SCEs), and micronucleus (MN) tests. The cells were treated with 13, 26 and 52 μg/mL 4-thujanol in the presence and absence of a metabolic activator (S9 mix). 4-Thujanol induced CA (P < 0.001) and MN formation (P < 0.05) at all concentrations (13, 26 and 52 μg/mL) in the presence and absence of the S9 mix without a concentration-dependent manner. However, the treatment of peripheral lymphocytes with 4-thujanol did not produce a statistical difference in the frequency of SCEs when compared with control group. Furthermore, this monoterpene did not significantly decrease the mitotic index (MI), proliferation index (PI), and nuclear division index (NDI). In conclusion, 4-thujanol had a significant clastogenic effect at the tested concentrations (13, 26 and 52 μg/mL) for human PBLs. In addition, no cytotoxic and/or cytostatic effects were observed regardless of the concentrations used. This work presents the first report on genotoxic properties of 4-thujanol.     

In vitro cytotoxic potential of newly synthesized furo[3,2-<Emphasis Type="Italic">c</Emphasis>]pyran-4-one derivatives in cultured human lymphocytes

The in vitro cytotoxic potentials of Furo[3,2-c]pyran-4-one derivatives in human lymphocytes were investigated. Blood samples were obtained from six healthy donors, non-smoking volunteers, which were incubated and exposed to increasing concentrations (0.05, 0.1, 0.5, 1 and 2 mg/mL) of Furo[3,2-c]pyran-4-one derivatives which are methyl 2-methoxy-7-(4-methylbenzoyl)-6-(4-methylphenyl)-4-oxo-4H-furo[3,2-c]pyran-3-carboxylate (1a) and methyl 2-methoxy-7-(4-methoxybenzoyl)-6-(4-methoxyphenyl)-4-oxo-4H-furo[3,2-c]pyran-3-carboxylate (1b). Compounds 1a and 1b induced micronucleus, mitotic and replication indexes in human lymphocytes (1 and 2 mg/mL). The increases of micronucleus, mitotic and replication indexes show that compounds at high concentrations may become cytotoxic, genotoxic and carcinogenic.     

Cytological effects of papaverine hydrochloride on root tips ofAllium cepa L.

Papaverine hydrochloride, which is an alkaloid found in opium, inducedc-metaphase,c-anaphase, non-congression, diagonal anaphase, chromosome and chromatid bridges, lagging of chromosomes, micronuclei and multinucleated cells in root tips ofAllium cepa. It is the only one amongst the alkaloids of opium which caused a high increase in MI after a long period of treatment with some low concentrations.