Skip to main content
NCBI home page
Cytotechnology logoLink to Cytotechnology
. 2014 Nov 1;68(3):443–450. doi: 10.1007/s10616-014-9797-x

Induction of giant cells by the synthetic food colorants viz. lemon yellow and orange red

V Prajitha 1,✉, John E Thoppil 1
PMCID: PMC4846638  PMID: 25366067

Abstract

Cytotoxicity and giant cell formation induced by lemon yellow and orange red synthetic food colorants were evaluated in the present study. The aqueous solutions of both the dye solutions were tested for cytotoxicity using Allium cepa assay. Frequency of giant cells were determined after treating the root tips with different concentrations of both food colorant solutions viz., 0.005, 0.01, 0.05, 0.1 % for varying time durations (1/2, 1, 2, 3 h). These colorants may cause giant cell formation primarily by interfering with the normal course of mitosis. Giant cells showing multiple aberrations viz. bridged and binucleate condition, cellular fragmentation, nuclear lesion, double and multiple nuclear lesions, double nuclear peaks and cellular breakage, elongated nucleus, nuclear budding, hyperchromasia, micronucleus, nuclear erosion, pulverized nucleus etc. were induced in root tips treated with both of the colorants. The synthetic food colorant treated cells showed inhibition of cell division and induction of giant cells. A dose dependant decrease in the mitotic index [88.20 % (c−ve, 3h) to 81.54 % (Lx4, 3h) and 88.20 % (c−ve, 3h) to 73.17 % (Ox4, 3h)] was observed. All mitotic phases show significant induction of giant cells when treated with both food colorants. Interphase stage shows higher percentage of giant cells, whereas in cytokinesis it was negligible. The orange red food colorant is observed to be more toxic because it recorded higher percentage of giant cell induction when compared with lemon yellow [27.93 % (Lx4, 3h) and 28.07 % (Ox4, 3h)].

Keywords: Allium cepa, Cytotoxicity, Giant cell, Lemon yellow, Orange red, Synthetic food colorants

Introduction

Studies on the toxicity of synthetic food additives have immense relevance in today’s world, since they cause a number of dreadful diseases. Today human beings have been directly exposed to various chemical substances. Use of chemicals by man is increasing day by day either as preservatives or as coloring medium. Food colorants are dyes, pigments or substances that can impart color when added to food items. Synthetic food colorants are one of the major groups of synthetic food additives. Lemon yellow and orange red synthetic food colorants are the most commonly used food colorants. The excessive use of these colorants results in dreadful toxic effects in organisms. Uses of such chemicals are very specific and are cautioned under the prevention of food and adulteration act (section 28, 1955). The regular use of synthetic food colors in various foodstuffs is a major problem with respect to human health (Somesh et al. 2005). But people use different non-permissible chemical dyes indiscriminately in varying concentrations for personal benefits, since general public is not aware of the chemical composition of these dyes (Hossain et al. 2002).

Synthetic food additive colours have been introduced in our food in order to give an attractive colour to it. They are used in many forms such as liquids, powders, gels and pastes, in commercial food production and domestic cooking. The use of synthetic food additives in general and colours in particular has recently increased considerably. The criteria for the quality of food products are apart from microbiological aspects, generally based on the colour, flavor, texture and nutritive value. However, one of the most important sensory qualities of a food product is the colour and the synthetic dyes monitoring (Sorouraddin and Saadati 2010).

It has been noticed that most of the applied synthetic food colours have properties similar to the mutagenic chemicals. Evidences have accumulated in the last few years to indicate that a large number of synthetic food additives are capable of inducing genetic diseases to human (Miller et al. 1996; Karen et al. 2006; Martin 2007).

Repeated exposure to even the permitted synthetic colours may be hazardous. Many of these dyes are originally derived from coal tar, commonly called coal-tar dyes, containing the azo group. By definition, colored chemicals are active chemicals, requiring greater care than bland additives such as emulsifiers (Saleem et al. 2013).

For each of the permitted colours the joint FAO/WHO expert committee of food additives has set an acceptable daily intake (ADI). The acceptable daily intake (ADI) has been defined as the amount of a substance that can be consumed everyday throughout the lifetime of an individual without any appreciable health effects (JECFA 1995).

The formation of giant cell is one of the major consequence produced due to the indiscriminate use of synthetic food colorants. Giant cells are formed when cells attempting to undergo mitosis, fail to complete cytoplasmic division. They grow in size and undergo DNA replication and nuclear division before they die (Kenne et al. 1986). Earlier studies had proven that artificial yellow dye caused a decrease in mitotic index at all the concentrations tested and induced wide spectrum of chromosomal abnormalities (Hossain et al. 2002). Cytotoxic effects of many synthetic dyes have already been tested in life systems (Khanna and Mukul 1991; Kaur et al. 1993; Shukla and Datta 1999; Sudhakar et al. 2001). The present study envisages cytotoxic aspects of synthetic food colorants, lemon yellow and orange red using Allium cepa assay.

Materials and methods

Collection of test materials

The synthetic food colorants, lemon yellow containing tartrazine (CAS No. 1934-21-0) and orange red containing both carmoisine (CAS No. 3567-69-9) and sunset yellow (CAS No. 2783-94-0) were purchased in pure form from the local market.

Preparation of test solution

To make the stock solution, 1 g of both colorants was weighed and dissolved in 100 ml distilled water. Lowest concentrations of the dye solutions viz., 0.005, 0.01, 0.05, 0.1 % [Lemon yellow (Lx1, Lx2, Lx3, Lx4) and Orange red (Ox1, Ox2, Ox3, Ox4)] were prepared for toxicity analysis by diluting the stock solution with distilled water.

Distilled water and 0.01 % methyl parathion was taken as the negative (C−ve) and positive control (C+ve) respectively. The parameters studied included mitotic index and percentage of giant cells induced.

Giant cell formation

The induction of giant cells by synthetic food colorants, lemon yellow and orange red was screened using A. cepa root tip meristematic cells, which have been used extensively in the screening of drugs or various compounds with cytotoxicity or antimitotic activity.

Certified bulbs of A. cepa, purchased from agricultural vendor were germinated over water before being transferred to each of the test dye solutions. When the roots were about 5 mm long, the bulbs were placed on beakers containing the dye solutions of four different concentrations (0.005, 0.01, 0.05, 0.1 %), such that the roots were immersed in the dye solutions. The duration of treatments for each concentration of both dye solutions was ½, 1, 2 and 3 h. Bulbs with sprouted roots were also treated with negative (C−ve) and positive (C+ve) control. The experimental set up had three replicates. The root tips were harvested after the treatment duration, washed and fixed in Carnoy’s fluid (1 part of glacial acetic acid: 2 parts of absolute alcohol) for 1 h. Mitotic squash preparation was done with the help of improved techniques (Sharma and Sharma 1990). Hydrolysis with 1 N HCl for 5 min and staining with 2 % acetocarmine for 4 h were carried out.

All the slides were scanned under Magnus microscope, tabulated and photomicrographs were taken with an Olympus Camedia C-4000 zoom digital camera (Tokyo, Japan) and AmScope MU Series digital camera. The numbers of dividing and non-dividing cells as well as the number of giant cells at different mitotic phases were recorded. Incidence of chromosome aberrations was calculated by expressing the number of aberrant cells as a percentage of total dividing cells for each treatment. Mitotic index was calculated by expressing the number of dividing cells as a percentage of total cells counted for each of the treatments and the control. Mitotic index and abnormality percentage were calculated using the following formulae:

Mitoticindex=NumberofdividingcellsTotalnumberofcells×100
Abnormalitypercentage=Numberofabnormal(Giant)cellsTotalnumberofcells×100

Statistical analysis

Data obtained on mitotic index and chromosomal aberrations were subjected to statistical analysis. Duncan’s multiple range test and one way ANOVA was performed to determine mean separation and significance of treatments using SPSS version 20, SPSS Inc.(Chicago, IL, USA). Data were expressed as mean ± standard error of mean (SEM). p < 0.05 was considered to be statistically significant.

Results and discussion

The results revealed drastic toxic effects of both synthetic food colorants on A. cepa root tip meristematic cells. The synthetic food colorant treated cells showed inhibition of cell division (Table 1) and induction of giant cells (Table 2; Figs. 1, 2, 3). Treatment of A. cepa root meristem with lemon yellow and orange red synthetic food colorant solutions resulted in a decrease in mitotic index (Table 1). Mitotic index was found to be lower than C−ve and higher than C+ve. Mitotic index values were observed to be inversely proportional to concentration and duration of treatment of both dye solutions. Lowering of mitotic index is generally considered as the result of cytotoxic activity. A dose dependant decrease in the mitotic index [88.20 % (c−ve, 3 h) to 81.54 % (Lx4, 3 h) and 88.20 % (c−ve, 3 h) to 73.17 % (Ox4, 3 h)] was also observed. Hossain et al. (2002) have observed that the artificial yellow dye cause decline in mitotic index with increase in dye concentration and duration of treatment. The complete arrest of cell division in A. cepa root meristematic cells when treated with orange red synthetic food colorant was also observed by Joseph and Siril (2010).

Table 1.

Mitotic index in control and dye treatments

Treatment Time duration (h) Total cells Dividing cells Mitotic index ± SE
C−ve ½ 892 750 84.08 ± 0.53b
1 968 820 84.70 ± 0.58d
2 901 764 84.82 ± 0.61c
3 899 793 88.20 ± 0.58c
C+ve ½ 788 307 38.95 ± 0.58a
1 916 292 31.87 ± 0.58a
2 832 208 25.00 ± 0.57a
3 865 189 21.84 ± 0.58a
Lx1 ½ 894 750 83.89 ± 0.58b
1 843 707 83.86 ± 0.58c,d
2 902 752 83.37 ± 0.58b,c
3 894 745 83.33 ± 0.58b
Lx2 ½ 926 770 83.15 ± 0.58b
1 898 746 83.08 ± 0.57b,c,d
2 910 755 82.96 ± 0.58b,c
3 900 746 82.88 ± 0.58b
Lx3 ½ 865 714 82.88 ± 0.58b
1 956 788 82.42 ± 0.58b,c
2 1,079 888 82.29 ± 0.58b
3 862 709 82.25 ± 0.58b
Lx4 ½ 719 591 82.19 ± 0.58b
1 887 725 81.73 ± 0.58b
2 1,127 920 81.63 ± 0.58b
3 932 760 81.54 ± 0.58b
Ox1 ½ 967 788 81.48 ± 0.58c
1 925 752 81.29 ± 0.58c
2 887 729 81.18 ± 0.58c
3 953 772 81.03 ± 0.55d
Ox2 ½ 857 694 80.98 ± 0.58c
1 1,023 826 80.74 ± 0.58c
2 883 713 80.74 ± 0.58c
3 1,075 864 80.37 ± 0.58c,d
Ox3 ½ 906 728 80.35 ± 0.58c
1 819 658 80.34 ± 0.58c
2 935 749 80.10 ± 0.58c
3 964 762 79.04 ± 0.58c
Ox4 ½ 867 680 78.43 ± 0.58b
1 702 549 78.21 ± 0.58b
2 821 636 77.46 ± 0.58b
3 820 600 73.17 ± 0.58b
Open in a new tab

C−ve, negative control (distilled water); C+ve, positive control (0.01 % methyl parathion); Lx1, Lx2, Lx3, Lx4, lemon yellow concentrations 0.005, 0.01, 0.05, 0.1 %, respectively; Ox1, Ox2, Ox3, Ox4, orange red concentrations 0.005, 0.01, 0.05, 0.1 %, respectively; SE, standard error. Means within a column followed by the same letters are not significantly different (p < 0.05) as determined by DMRT

Table 2.

Induction of giant cells in control and dye treatments

Treatment Time duration (h) Number of giant cells at different phases % of giant cells ± SE
I P M A T
C +ve ½ 60 40 40 40 – 22.84 ± 0.58d
1 80 20 30 20 33 23.47 ± 0.58d
2 35 40 25 40 60 25.38 ± 0.58d
3 80 60 10 60 10 27.91 ± 0.58d
Lx 1 ½ 39 35 16 20 8 13.42 ± 0.58a
1 42 34 14 10 8 13.65 ± 0.33a
2 36 45 15 13 19 14.12 ± 0.58a
3 41 40 23 22 – 14.31 ± 0.58a
Lx 2 ½ 52 49 28 12 – 15.22 ± 0.58b
1 48 54 29 14 – 16.14 ± 0.58b
2 63 48 31 – 10 17.02 ± 0.58b
3 52 52 41 20 – 18.33 ± 0.58b
Lx 3 ½ 63 12 39 22 30 19.19 ± 0.58c
1 75 48 25 33 14 20.39 ± 0.58c
2 81 63 44 42 – 21.31 ± 0.58c
3 85 40 27 31 8 22.15 ± 0.58c
Lx 4 ½ 40 20 63 25 18 23.11 ± 0.55d
1 51 79 28 31 20 23.56 ± 0.58d
2 79 65 69 72 4 25.64 ± 0.58d
3 74 55 42 54 35 27.93 ± 0.58d
Ox 1 ½ 44 30 21 20 15 13.96 ± 0.58a
1 40 29 28 22 12 14.91 ± 0.58a
2 42 33 31 29 12 16.23 ± 0.67a
3 47 41 34 25 8 16.78 ± 0.58a
Ox 2 ½ 50 47 30 31 9 16.78 ± 0.58a
1 58 51 42 38 11 19.55 ± 0.58b
2 51 38 40 21 18 19.93 ± 0.58b
3 59 42 51 31 28 20.46 ± 0.58b
Ox 3 ½ 69 51 45 17 13 21.52 ± 0.58c
1 67 46 38 20 10 22.58 ± 0.58c
2 69 59 40 32 20 23.52 ± 0.58c
3 63 51 50 41 16 23.85 ± 0.58c
Ox 4 ½ 64 52 39 34 21 24.22 ± 0.58d
1 64 48 29 10 14 24.92 ± 0.58d
2 72 55 41 – 42 25.57 ± 0.58d
3 70 49 43 29 39 28.07 ± 0.55d
Open in a new tab

C+ve, positive control (0.01 % methyl parathion); Lx1, Lx2, Lx3, Lx4, lemon yellow concentrations 0.005, 0.01, 0.05, 0.1 %, respectively; Ox1, Ox2, Ox3, Ox4 , orange red concentrations 0.005, 0.01, 0.05, 0.1 %, respectively; I, interphase, P, prophase, M, metaphase, A, anaphase, T, telophase; SE, standard error. Means within a column followed by the same letters are not significantly different (p < 0.05) as determined by DMRT

Fig. 1.

Fig. 1

Open in a new tab

Graph showing induction of giant cells at different mitotic phases and varying concentrations of lemon yellow dye solution. I interphase, P prophase, M metaphase, A anaphase, T telophase, Lx lemon yellow

Fig. 2.

Fig. 2

Open in a new tab

Graph showing induction of giant cells at different mitotic phases and varying concentrations of orange red dye solution. I interphase, P prophase, M metaphase, A anaphase, T telophase, Ox orange red

Fig. 3.

Fig. 3

Open in a new tab

Giant cells induced by synthetic food colorants showing various cytological aberrations. a Bridged and binucleate cell, b cellular fragmentation, c nuclear lesion; d double nuclear lesion, e double nuclear peaks and cellular breakage, f elongated nucleus, g nuclear budding, h hyperchromasia, i micronucleus, j multiple nuclear lesion, k nuclear erosion, l pulverized nucleus, m unequal condensation in early prophase, n pole to pole metaphase arrangement with a fragment, o nuclear peaks at cytokinesis, p macro and microcell formation. Bar 10 µm

Cell division was found to be normal in the root tips treated with negative control. The giant cell induction scored slightly higher values [27.93 % (Lx4, 3 h) and 28.07 % (Ox4, 3 h)] in treatments when compared with C+ve [27.91 % (3 h)].

Formation of giant cell was the most frequent abnormality observed in both treatments. It may be due to the interference of food colorants with the normal course of mitosis. The number of giant cells was also observed to be increasing with the concentration and duration of treatment of the aqueous dye solutions. Giant cells containing multiple aberrations observed include bridged and binucleate condition, cellular fragmentation, nuclear lesion, double nuclear lesion, double nuclear peaks and cellular breakage, elongated nucleus, nuclear budding, hyperchromasia, micronucleus, multiple nuclear lesions, nuclear erosion, pulverized nucleus, unequal condensation in early prophase, pole to pole metaphase arrangement with a fragment, nuclear peaks at cytokinesis, macro and microcell formation, etc. (Fig.  3a–p).

The active ingredient of the synthetic food colorant lemon yellow is tartrazine [E102] (Gao et al. 2011) and that of orange red is carmoisine [E122] (Sorouraddin and Saadati 2010) and sunset yellow [E110] (Atri et al. 2014). These chemicals belong to azo group of food colorants, and are metabolized into aromatic amines, which can generate reactive oxygen species by interaction with nitrate or nitrite containing substrates in the cells (Amin et al. 2010). The reactive oxygen species (ROS) such as superoxide anion, hydroxyl radical and H2O2 could be produced in the metabolism of nitrosamines and increase oxidative stress (Bansal 2005). Thus the ROS generated may oxidize and damage lipids, proteins and DNA and lead to overall cytotoxic effects (Bakkali et al. 2008). The immunosuppressive effect of tartrazine was also reported by Koutsogeorgopoulou et al. (1998).

Inhibition of cytokinesis following telophase is responsible for binucleated cell formation (Majewska et al. 2003). Earlier reports suggest that the presence of nuclear lesions offer cytological evidence for the inhibitory action on DNA biosynthesis (Akaneme and Iyioke 2008; Mercykutty and Stephen 1980). Pulverized nucleus may arise due to the premature condensation of chromosome (Sakari et al. 1981). Progressive heterochromatinization seems to be the reason for hyperchromasia (Gernand et al. 2005). Nuclear buds arise as a result of the excessive production of nucleic acids and proteins, induced by the cytotoxicants (Hellgren and Morre 1992). Micronucleus may originate from a lagging chromosome at anaphase or from a chromosome fragment (Badr and Ibrahim 1987).

All mitotic phases show significant induction of giant cells when treated with both of the food colorants, of which interphase shows higher percentage and cytokinesis shows negligible rate. The orange red synthetic food colorant is observed to be more toxic because it recorded higher percentage of giant cell induction when compared with lemon yellow. Both of these colorants shows higher percentage of giant cell formation than C+ve when the root tips were treated with 0.1 % of dye solutions for 3 h. One way ANOVA showed that there was a significant effect of treatment on mitotic activity. Post-hoc analysis using Duncan’s multiple range test showed that the activity of both of the dyes were significant (p < 0.001) when compared with that of control.

Giant cells are formed when cells attempting to undergo mitosis fail to complete cytoplasmic division. They grow in size and undergo DNA replication and nuclear division before they die (Kenne et al. 1986). The giant cells induced by artificial yellow dye were also observed by Hossain et al. (2002). Mpountoukas et al. (2010) has also observed a decrease in mitotic index and increase in genotoxicity when certain food colorants were studied on human peripheral blood cells in vitro.

According to Cross et al. (1995), Fukasawa et al. (1996) and Morgan et al. (1996) multinucleate giant cell is formed because of the blocking of karyokinesis after replication, resulting in the formation of tetraploid giant cell. Some of the aberrations seemed to be formed during the last mitotic division cycle since no replicated chromosome aberrations were seen in these giant cells.

Giant cells can be induced either by physical means or by employing chemicals, which are capable of affecting the cell cycle especially in the ‘S’ phase. Here the cell division may be partially arrested due to the toxic effects of food colorants and it may be the reason for the reduction in mitotic index in treatments, followed by mitotic arrest. Cell expansion seems to be induced and as a result the cells become large giant cells. The frequency of giant cells can be increased depending upon the dosage and duration of the treatment with cytotoxic agents (Verma and van Huyste 1971). Moreover, it had been experimently proven that giant cells are subjected to various kinds of stress and normally they cannot cope with the enormous size of the protoplasm thus they are liable for injury (Menzel 1988). Joseph and Siril (2010) also observed the frequent occurrence of giant cells after Allium root tips having been treated with orange red synthetic.

X ray-irradiation can also interfere with the highly ordered sequence of events in cell division which affects cell cycle progression and hence causes giant cell formation (Baumstark-Khan et al. 1986). Spontaneous premature condensation, probably due to radiation-induced cell cycle delays the late S and G2 phases enhances the intracellular levels of cyclins B1 and may play a role in the mechanism underlying radiation-induced giant cells (Ianzini and Mackey 1997). Giant cell formation in human embryo cells after X-ray irradiation was observed by Roy et al. (1999). Crocidolite and chrysotile induced giant cell formation was reported by Hong and Choi (1997). The multinucleate giant cell was known to be associated with the final common pathway of germinal cell disintegration in animals treated with different chemicals (D’Souza 2003). Administration of sodium fluoride together with aluminium chloride on adult male mice for 30 days resulted in structural alterations in the testis with formation of giant cells (Chinoy et al. 2005).

The present study clearly indicates that both of these synthetic dyes have genotoxic effects on living system. It warrants the danger of illegal and indiscriminate use of such dyes in food and other consumable items. Chung et al. (1978) reported that azo dyes such as tartrazine and sunset yellow used widely can be reduced to aromatic amines by the intestinal microflora and hence cause intestinal cancer. An earlier report stated that the metabolism of azo dyes derived from benzidine when converted to aromatic amines by intestinal bacteria is potentially carcinogenic (Combes and Haveland-Smith 1982).

Mitotic interference induced by food colorants physically may be closely related with carcinogenicity. The giant cell containing multiple aberrations indicate that both the food colorants, especially at higher concentrations, are capable of causing changes in chromosome number and structure. Chromosome aberrations were observed in all stages of mitosis. The abnormalities of chromosomes could be due to the blockage of DNA synthesis or inhibition of spindle formation.

In summary, giant cells can be useful in evaluating the effects of synthetic food colorants on animal and human cells since multinucleate giant cells can easily be counted and the ability to induce formation of giant cells reflects the ability to interfere with mitosis. This index of mitotic disturbances seems to be a useful marker in the carcinogenesis assay of synthetic food colorants. The toxicity of these synthetic food colorants recorded in the present study needs follow-up research including further tests using cell lines and animal systems.

Acknowledgments

P. V. kindly acknowledges Kerala State Council for Science, Technology and Environment for providing financial assistance through KSCSTE fellowship.

References

  1. Akaneme FI, Iyioke IV. Mutagenic potentials of the sterilizing fluid-purital on root tip mitosis of Allium cepa. Bio-Research. 2008;6:293–297. [Google Scholar]
  2. Amin KA, Hameid HA, Elsttar AHA. Effect of food azo dyes tartrazine and carmoisine on biochemical parameters related to renal, hepatic function and oxidative stress biomarkers in young male rats. J Food Chem Toxicol. 2010;48:2994–2999. doi: 10.1016/j.fct.2010.07.039. [DOI] [PubMed] [Google Scholar]
  3. Atri R, Singh A, Mathur N, Verma A. Utilization of microbial bioassays for screening the possible toxicity in regularly used food dyes. J Chem Bio Phy Sci Sec B. 2014;4:1248–1257. [Google Scholar]
  4. Badr A, Ibrahim AG. Effect of herbicide ‘Glean’ on mitosis, chromosomes and nucleic acids in Allium cepa and Vicia faba root tip meristems. Cytologia. 1987;52:293–302. doi: 10.1508/cytologia.52.293. [DOI] [Google Scholar]
  5. Bakkali F, Averbeck S, Averbeck D, Idaomar M. Biological effects of essential oil—a review. J Food Chem Toxicol. 2008;46:446–475. doi: 10.1016/j.fct.2007.09.106. [DOI] [PubMed] [Google Scholar]
  6. Bansal AK. Modulation of N-nitrosodiethylamine induced oxidative stress by vitamin E in rat erythrocytes. Human Exp Toxicol. 2005;24:297–302. doi: 10.1191/0960327105ht533oa. [DOI] [PubMed] [Google Scholar]
  7. Baumstark-Khan C, Rink H, Zimmermann HP. Radiation induced formation of giant cells in Saccharomyces uvarum III. Effect of X-rays on nuclear division. Radiat Environ Biophys. 1986;25:23–30. doi: 10.1007/BF01209681. [DOI] [PubMed] [Google Scholar]
  8. Chinoy NJ, Sorathia HP, Jhala DD. Fluoride+aluminium induced toxicity in mice testis with giant cells and its reversal by vitamin C. Fluoride. 2005;38:109–114. [Google Scholar]
  9. Chung KT, Fulk GE, Egan M (1978) Reduction of azo dyes by intestinal anaerobes. Appl Environ Microbiol 35:558–562 [DOI] [PMC free article] [PubMed]
  10. Combes RD, Haveland-Smith RB. A review of the genotoxicity of food, drug and cosmetic colors and other azo, triphenylmethane and xanthene dyes. Mutat Res. 1982;98:101–248. doi: 10.1016/0165-1110(82)90015-X. [DOI] [PubMed] [Google Scholar]
  11. Cross SM, Sanchez CA, Morgan CA, Schimke S, Ramel RL, Idzerda WH, Raskind WH, Reid BJ. A p53-dependent mouse spindle checkpoint. Science. 1995;267:1353–1356. doi: 10.1126/science.7871434. [DOI] [PubMed] [Google Scholar]
  12. D’Souza UJA. Tamoxifen induced multinucleated cells (symplasts) and distortion of seminiferous tubules in rat testis. Asian J Androl. 2003;5:217–220. [PubMed] [Google Scholar]
  13. Fukasawa K, Choi T, Kuriyama R, Rulong S, Woude GFV. Abnormal centrosome amplification in the absence of p53. Science. 1996;271:1744–1747. doi: 10.1126/science.271.5256.1744. [DOI] [PubMed] [Google Scholar]
  14. Gao Y, Li C, Shen J, Yin H, An X, Jin H. Effect of food azo dye tartrazine on learning and memory functions in mice and rats, and the possible mechanisms involved. J Food Sci. 2011;76:125–129. doi: 10.1111/j.1750-3841.2011.02267.x. [DOI] [PubMed] [Google Scholar]
  15. Gernand D, Rutten T, Varshney A, Rubtsova M, Prodanovic S, Bruss C, Kumlehn J, Matzk F, Houben A. Uniparental chromosome elimination at mitosis and interphase in wheat and pearl millet crosses involves micronucleus formation, progressive heterochromatinization and DNA fragmentation. Plant Cell. 2005;17:2431–2438. doi: 10.1105/tpc.105.034249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hellgren L, Morre DJ. ATP induced budding of nuclear envelope in vitro. Protoplasma. 1992;167:238–242. doi: 10.1007/BF01403387. [DOI] [Google Scholar]
  17. Hong YC, Choi SS. Cytotoxicity and multinucleate giant cell formation in Chinese hamster lung fibroblast caused by crocidolite and chrysotile. J Korean Med Sci. 1997;12:99–104. doi: 10.3346/jkms.1997.12.2.99. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hossain Z, Shukla R, Mandal AKA, Datta SK. Allium test for assessing chromotoxic effects of artificial yellow dye. Cytologia. 2002;67:411–415. doi: 10.1508/cytologia.67.411. [DOI] [Google Scholar]
  19. Ianzini F, Mackey MA. Spontaneous premature chromosome condensation and mitotic catastrophe following irradiation of HeLa S3 cells. Int J Radiat Biol. 1997;72:409–421. doi: 10.1080/095530097143185. [DOI] [PubMed] [Google Scholar]
  20. Joint Expert Committee on Food Additives (JECFA). (1995) Summary of evaluations performed by the Joint FAO/WHO Expert Committee on Food Additives. 44th meeting, 14–23 February 1995. Geneva (Switzerland): FAO/IPCS/WHO, Geneva, pp 1956–1996
  21. Joseph N, Siril EA. Cytotoxic evaluation of Annatto (Bixa orellana L.) dye compared with orange red. Cytologia. 2010;75:163–167. doi: 10.1508/cytologia.75.163. [DOI] [Google Scholar]
  22. Karen L, Mclean WG, Williams DP, Howard CV. Synergistic interactions between commonly used food additives in a developmental neurotoxicity test. Toxicol Sci. 2006;46:177–184. doi: 10.1093/toxsci/kfj073. [DOI] [PubMed] [Google Scholar]
  23. Kaur A, Singh R, Grover IS. Mutagenicity testing of textile dyes with Salmonella microsome assay. J Environ Biol. 1993;14:327–333. [Google Scholar]
  24. Kenne K, Ljungquist S, Ringertz NR. Effects of asbestos fibers on cell division, cell survival and formation of thioguanine resistant mutants in Chinese hamster ovary cells. Environ Res. 1986;39:448–464. doi: 10.1016/S0013-9351(86)80069-X. [DOI] [PubMed] [Google Scholar]
  25. Khanna SK, Mukul D. Toxicity, carcinogenic potential and clinicoepidemiological studies on dyes and dye intermediates. J Sci Ind Res. 1991;50:965–974. [Google Scholar]
  26. Koutsogeorgopoulou L, Maravelias C, Methenitou G, Koutselinis A. Immunological aspects of the common food colorants, amaranth and Tartrazine. Vet Hum Toxicol. 1998;40:1–4. [PubMed] [Google Scholar]
  27. Majewska AE, Wolska E, Sliwinska M, Furmanowa N, Urbanska A, Pietrosiuk A, Zobel A, Kuras M. Antimitotic effect, G2/M accumulation, chromosomal and ultrastructure changes in meristematic cells of Allium cepa L. root tips treated with the extract from Rhodiola rosea roots. Caryologia. 2003;56:337–351. doi: 10.1080/00087114.2003.10589343. [DOI] [Google Scholar]
  28. Martin H (2007) Soft drinks cause degenerative DNA diseases. http://news.indep endent.Co.uk/healtharticle
  29. Menzel D. How do giant plant cells cope with injury? The wound response in siphonous green algae. Protoplasma. 1988;144:79–91. doi: 10.1007/BF01637240. [DOI] [Google Scholar]
  30. Mercykutty VC, Stephen J. Adriamycin induced genetic toxicity as demonstrated by the Allium test. Cytologia. 1980;45:769–777. doi: 10.1508/cytologia.45.769. [DOI] [Google Scholar]
  31. Miller ME, Lummus ZL, Bernstei DI. Occupational asthma caused FD8 and C blue dye No. 2. Allergy Asthma Proc. 1996;17:31–34. doi: 10.2500/108854196778662309. [DOI] [PubMed] [Google Scholar]
  32. Morgan WF, Day JP, Kaplan MI, Mc Ghee EM, Limoli CL. Genomic instability induced by ionizing radiation. Radiat Res. 1996;146:247–258. doi: 10.2307/3579454. [DOI] [PubMed] [Google Scholar]
  33. Mpountoukas P, Pantazaki A, Kostareli E, Christodoulou P, Kareli D, Poliliou S, Mourelatos C, Lambropoulou V, Lialiaris T. Cytogenetic evaluation and DNA interaction studies of the food colorants amaranth, erythrosine and tartrazine. J Food Chem Toxicol. 2010;48:2934–2944. doi: 10.1016/j.fct.2010.07.030. [DOI] [PubMed] [Google Scholar]
  34. Roy K, Kodama S, Suzuki K, Watanabe M. Delayed cell death, giant cell formation and chromosome instability induced by X-irradiation in human embryo cells. J Radiat Res. 1999;40:311–322. doi: 10.1269/jrr.40.311. [DOI] [PubMed] [Google Scholar]
  35. Sakari K, Martti S, Pekka V. Chromosome pulverization in blood diseases. Hereditas. 1981;95:15–24. doi: 10.1111/j.1601-5223.1981.tb01323.x. [DOI] [PubMed] [Google Scholar]
  36. Saleem N, Umar ZN, Khan SI. Survey on the use of synthetic food colors in food samples procured from different educational institutes of Karachi city. J Trop Life Sci. 2013;3:1–7. [Google Scholar]
  37. Sharma AK, Sharma A. Chromosome techniques-theory and practice. 3. London: Butterworths; 1990. [Google Scholar]
  38. Shukla R, Datta SK. Cytotoxic effects of artificial blue dye. J Cytol Genet. 1999;34:95–97. [Google Scholar]
  39. Somesh Y, Meenu S, Kumar SH. Evaluation of cytotoxicity of some synthetic food colours. Int J Plant Res. 2005;18:55–65. [Google Scholar]
  40. Sorouraddin MH, Saadati M. A simple fabrication of portable diffuse reflectometer for simultaneous analysis of common food dyes. Sensor Actuat B-Chem. 2010;145:367–372. doi: 10.1016/j.snb.2009.12.036. [DOI] [Google Scholar]
  41. Sudhakar R, Gowda KNN, Venu G. Mitotic abnormalities induced by silk dyeing industry effluents in the cells of Allium cepa. Cytologia. 2001;66:235–239. doi: 10.1508/cytologia.66.235. [DOI] [Google Scholar]
  42. Verma DPS, van Huyste RB. Induction of giant cells in suspension cultures of Arachis hypogea L. by massive irradiation. Radiat Res. 1971;48:518–530. doi: 10.2307/3573338. [DOI] [PubMed] [Google Scholar]

Articles from Cytotechnology are provided here courtesy of Springer Science+Business Media B.V.

RESOURCES