Nonlinear optical Zinc mixed bisthiourea Cadmium chloride (BTCC) crystals ... Experimental Growth of Pure BTCC. Crystals. BTCC crystal was synthesized by dissolving AR grade ..... Borwick, âSalt-Based Approach for Frequency Conver-.
curative treatment of various cancers such as, testicular teratoma ... and neck and testicular cancers. ..... mutagenicity in mice treated with niclosamide. Mutation ...
Ni per rat) did not alter the blood hematocrit or reticulocyte count during six weeks of observation. .... search Laboratory, INCO Ltd., Clarkson, Ontario,. Canada.
no nickel in MT-fractions obtained by gel-filtration chromatography of hepatic cytosol from Ni[II]-treated rats (table II). Sarkar24-25 reported that 63Ni in renal cy.
ISSN 0895-8378 print/ISSN 1091-7691 online. DOI: 10.3109/08958378.2011.617398. Inhalation Toxicology Downloaded from informahealthcare.com by Cukurova Univ. on ..... and the regulation of neuronal signalling. Trends Pharmacol Sci. 25:317â324. DiFra
Jul 1, 2013 - mass spectrometry (ICP-MS) was carried out to trace the distribution of ... Cyanobacteria; Nickel Chloride; Stress Response;. ICP-MS. 1. INTRODUCTION. The formation of algal blooms is a serious concern around the world. Algal blooms ...
Intestinal absorption of nutrients in rats treated with 2,3,7,8- tetrachlorodibenzo-p-dioxin TCDD. J Toxicol Environ;8:629â38. Bancroft, J.D., Stevens, A. 1986. Theory and Practice of Histological Techniques. Churchill-Livingstone, London. Bhattach
... Farmacologia e BiofÃsica, Instituto de BiociÃªncias, UFRGS, Av. Sarmento. Leite ..... Sei. USA, 75:6149-6153. KUNZ, B.; HANAN, M. A. & HA YNES, H., 1980.
nicotine strongly attenuated staurosporine (STP)-induced apopto- sis. ..... 5A and. 5B). Cultures pretreated with NAC or CAT for 2 h before nicotine addition had ...
Jun 29, 2011 - On hypernatremic challenge, the ... brain of chronic hypernatremic rats (Sterns and Silver, ... receiving lithium chloride (Parkinson et al, 2009).
This experiment was carried out to evaluation usage different levels of Choline Chloride Supplement ... with Duncan test. Three male Japanese quail selected with each pen and slaughtered. Result showed choline chloride supplement in all levels not si
Jul 2, 2013 - and LSD multiple comparison test (p < 0.05) using the SPSS statistical ..... Jaleel CA, Manivannan P, Sankar B, Kishorekumar A, Gopi R et al.
First principles studies of the effect of nickel carbide catalyst composition on carbon nanotube growth. Anders BÃ¶rjessonâ. University of BorÃ¥s, 501 90 BorÃ¥s, ...
Apr 11, 2017 - Effect of Potassium Chloride on the Blood Pressure in Two-Kidney ... In both studies, potassium loading increased fluid intake and urine volume, which were ..... Davis JO, Freeman RH: Mechanisms regulating renin release.
Nov 13, 2017 - Abstract: We analyzed the utility of ammonium chloride (NH4Cl) as a nitrogen source .... High-performance liquid chromatography (PerkinElmer.
comprising of fungicides like Chlorthalonil, Carbendazim, Fenarimol and Zineb, insecticides like Deltamethrin, Malathion, Malaoxon,. Isomalathion, Permethrin ...
faba bean: new loci from Orobanche-resistant cultivar. "Giza 402". Plant Breeding, 128-155. Zienolddiny, S., D.H. Svendsrud, D. Ryberg, A.B. Mikalsen and.
The effect of heavy metal nickel chloride on germination, growth and biochemical parameters of Phaseolusmungo L. was studied. Application of various concentrations from 3mM to 15mM decreased the percentage of growth, pigment content and increased the
Mar 30, 2014 - www.iajpr.com. Page. 14. 4. 9. Indo American Journal of Pharmaceutical Research, 2014. ISSN NO: 2231-6876. Journal home page:.
trations showed that at low chloride ion concentration. (5 x. 10e3 M) the alkaline ... ber of protons bound by Hb and HbOe by (3) : a log P50 = 1 a PH. qAZ B. (1).
Brown cows from a farm close to and in front of the Skrunda Radar and from ... lymphocytes induced by 1.8 GHz radiofrequency field radiation (RFR, SAR of ...... to 2450 MHz microwaves during 30 and 120 min at a constant temperature of 36.1.
Aug 11, 2014 - ... and so on, and can accumulate in plants, animals or soil, which can affect the environment and even ...... Ayvaz, Z. Cevre Kirliligi ve Kontrolü.
alytic hydrogenation of nitrobenzene,1â3 and an electrochemical synthesizing method.4 The major disadvantage of the iron-acid reduction is the gener- ation of ...
Studies on the Genotoxic Effect of Nickel Chloride in Mice and the PossibleProtective Role of Soybean Seeds Extracts 1
Maha A. Fahmy, 2Nagwa H.A. Hassan, 3Farouk R. Melek, 3 Zeinab M. Hassan and 3Hanan A. Al-Ashaal
Department of Genetics and Cytology, National Research Centre, Dokki, Cairo, Egypt 2 Faculty of Science Ain Shams University, Cairo, Egypt 3 Department of Chemistry of Natural Compounds, National Research Centre, Dokki, Cairo, Egypt
Abstract: Nickel is an essential trace element. Nickel and its compounds have many industrial applications. Concern about nickel compounds is due to potential occupational and environmental health hazards. In the present study the mutagenic potential of nickel chloride (NiCl 2) was tested to evaluate somatic and heritable disorders which may extend to the next generations and associated with increased risk of cancer. The protective role of dietary soybean seeds and its extracts was also evaluated. The study examined the induction of chromosomal aberrations in bone marrow cells and mouse spermatocytes and the effect on morphological sperm abnormalities. The results showed that NiCl2 induced an increase in the percentage of chromosomal aberrations in somatic and germ cells with dose and time relationship. A significant (P<0.01) increase in the percentage of abnormal sperm was observed after treatment with the tested doses. The results also demonstrated that soybean seeds and its extracts have a protective role and reduced the mutagenic potential induced by NiCl 2. Methanol extract and methanol fraction of soybean seeds gave the most effective protection. Key words: Nickel Chloride Mutagenicity Role Soybean Seeds Extracts
High quantity of nickel has been reported to show various toxicities such as pulmonary, renal and cardiovascular effects [2-4]. Carcinogenic and mutagenic potential of nickel were demonstrated . Nickel can cross the placental barrier, affecting directly the developing embryo or fetus in experimental animals . It affects various aspects of reproduction  and has immunosuppressive effect . Some nickel compounds have been established as human carcinogens based on epidemiological evidence, which show high incidence of nasal and lung cancers in refinery workers  and its potency to induce tumors in a variety of mammalian species . The carcinogenicity of Ni compounds is believed to be associated with their solubility and cellular uptake. Water insoluble Ni compounds tend to be carcinogenic than Ni soluble compounds because they are phagocytize by cells resulting in intracellular delivery of high quantities of Ni . However, the carcinogenicity of a compound is
Nickel is a natural element of the earth’s crust; therefore small amounts are found in food, water, soil and air. Individuals also may be exposed to nickel in occupations involved in its production, processing and use, or through contact with every item such as nickel containing jewelry and stainless steel cooking and eating utensils and by smoking tobacco. Smoking 40 cigarettes daily, for example, may result in inhalation of 2-23µ g of nickel. Nickel is found in ambient air at very low levels as a result of releases from oil and coal combustion, west and sewage sludge incineration, nickel metal refining, manufacturing facilities and other sources . Nickel (Ni+2) in the body is an essential trace element required in micro quantity. Nickel plays some important role in biological system such as enzyme activity, hormonal control and also in DNA, RNA and protein structure and function .
Corresponding Author: Maha A. Fahmy, Department of Genetics and Cytology, National Research Centre, Dokki, Cairo, Egypt .
Global J. Pharmacol., 8 (4): 625-634, 2014
closely related to its genotoxic and mutagenic potential. In mammalian cells, the induction of sister chromatid exchanges and chromosomal aberrations, DNA-single strand breaks and DNA- protein cross-links were demonstrated with various nickel salts [12-15]. Nowadays, the beneficial diets are of particular importance since they may have a role in the prevention against many diseases and cancer. Soybean (Glycine max) is one of the major agronomic crops in many countries. Soybean containing up to 40% protein can be considered as a protein concentrate even without defate. In comparison with other plant proteins, soybean protein is lysine-rich and is therefore useful as a supplement to cereals and also as low cost quality protein for human meals. Soybean is highlighted now as a source of pharmaceutical raw materials and drugs. Various soybean contents are proved to have immune-improving, anticancer, antimutagenic effects and are potent against a broad spectrum of diseases [16, 17]. The present study concerning with, evaluating the genotoxic effects of NiCl2 in somatic and germ cells of male mice, detecting the possible protective role of soybean seeds extracts and correlating this activity with the chemical composition of these extracts. The study assumes importance in view of widespread human exposure to nickel compounds.
It is noteworthy that the average food intake of each mouse (25g) is 4.5±0.5g/day. In groups of experimental animals which feed soybean seeds, the seeds were mixed as 30% with the solid diet of animals under test. So,1.5g of soybean seeds are required/mouse/day. This amount of food intake from soybean was taken into consideration as bases for calculating the doses of soybean seeds extracts in all experiments. Soybean extracts was prepared as follows: Soybean seeds (2Kg) were covered with methanol and left for three days at room temperature. The process of extraction was repeated 3 times. The solvent from combined methanol extract was evaporated under reduced pressure. The residue obtained (159.9g) was dissolved in dist water at the desired concentration just before use (Methanol Extract- M.E. Another batch of soybean seeds (2kg) was extracted with n-hexane to give 266.6g of n-hexane fraction (n-hx.F.) after evaporation and dryness and used in cytogenetic experiments. For preparation of chloroform fraction (Ch.F.) of soybean, the plant material after extraction with n-hexane was left to dry and then extracted with chloroform (3 times). The residue obtained after dryness (23.5g) was used in cytogenetic study after dissolving in corn oil at the desired concentration just before use. The plant material, after extraction first with n-hexane, then with chloroform was left to dry. The dried material was then extracted with methanol (3 times) to give the methanol fraction of soybean seeds(M.F.).The obtained residue was weighted (2Kg gave 173.3g ) and dissolved in dist water at the desired concentration just before use. Isolation and Identification of active constituents were performed,described and published in Al-Ashaal et al. 
MATERIAL AND METHODS Animals: Male white Swiss mice ( Mus Musculus ), aged 9-12 weeks were used in all experiments.The ethical approval for the experiments was obtained from the ethical committee of the National Research centre, Cairo, Egypt. The animals were obtained from a closed random bred colony at the National Research Centre. The mice used for any one experiment were selected from mice of similar age (± 1 week) and weight (± 2g). Animals were housed in polycarbonate boxes with steel-wire tops (not more than five animals/ cage) and bedded with wood shavings. Ambient temperature was controlled at 22± 3°C with a relative humidity of 50 ± 15% and a 12- h light/dark photoperiod. Food and water were provided ad libitum.
Treatment and Cytological Preparations: Chromosomal Aberration: Single-dose Treatment: Mice received a single i.p treatment with NiCl2 at dose levels 2.625, 5.250, 10.50 and 21.00 mg Kg 1b,wt (equivalent to 1/16, 1/8, 1/4 and ½ of the experimental LD50 ).
Chemicals: Nickel chloride (NiCl2) anhydrous was purchased from Merck-Schuchardt Co. Germany.
Repeated -Dose Treatment: Mice received a daily i.p treatment with one of the doses 2.625, 5.250 and 10.50 mg Kg 1 bwt NiCl2 for 1,2 and 3 weeks and feed normal diet. With respect to the dose 5.250 mg Kg 1bwt NiCl2 (1/8LD50), in addition to NiCl2-treated groups, also the effect of (NiCl2 + M.E), (NiCl2 +n-hx.F.), (NiCl2 +Ch.F.) and (NiCl2 + M.F.) were tested. All extracts were given orally. Another groups of mice received the same dose of NiCl 2 but feed diet mixed with 30% soybean seeds.
Solvents for Extraction: n-hexane, chloroform, methanol. Plant Material and Extract Preparation: soybean seeds obtained from the Agriculture Research Centre (Egypt) were used. The seeds were autoclaved at 107°C for 15 min, let to cool then it were ground in an electric grinder. 626
Global J. Pharmacol., 8 (4): 625-634, 2014
Slide Preparation and Scoring: Mice were i.p injected with colchicine 2-3h before sacrificied. Bone-marrow preparations were made according to the technique described by Yosida and Amano . A group of five mice was used for each treatment and 75 well spread metaphases were analyzed/animal for scoring different types of abnormalities. Chromosomal preparations from testes were made according to the technique developed by Evans et al . 100 well-spread diakinesis metaphase I cells were analyzed per animal to assess abnormalities in five mice/group. Scoring was performed under (25×100) X magnification with a light microscope (Litz, Germany).
RESULTS Effect of Nickel Chloride: Chromosomal Aberration in Bone Marrow: The results in Tables (1,2) show that different doses induced significant percentage of chromosomal aberrations. Such percentage was found to be dose-and time dependent. The maximum percentage reached 28.26± 0.32 (P<0.01) 3 weeks after treatment with the dose 10.50 mgkg 1bwt NiCl2 compared with 4.5±0.32 for control. Different types of chromosome aberrations were recorded. Chromosomal Abnormalities in Spermatocytes: Nickel chloride at the tested doses induced a statistically significant percentage of chromosomal abnormalities in mouse spermatocytes (except 24h after single i.p treatment with the dose 2.625). This percentage increased with increasing the dose and with longer duration of treatment. The maximum percentage of abnormalities reached 17.00 ± 0.71 (P< 0.01) which represents about 4.8 fold increases compared with the negative control 3.60 ± 0.51. Nickel chloride affected separation in univalents, X-Y univalent dominated. Fragments/ breaks, gaps and Translocations as chain IV were observed (Tables 3, 4 )
Sperm -Shape Abnormalities: Groups of five mice each were i.p. treated with NiCl2 daily for five consecutive days at dose levels of 2.625, 5.250 and 10.50 mg Kg 1 b.wt and feed normal diet. Other groups of mice treated with the same doses but feed normal diet mixed with 30% soybean seeds. Animals were sacrificed 35 days after the first treatment by cervical dislocation. Sperm were prepared according to the recommended method of Wyrobek and Bruce . The epididymides were excised and minced in 2ml physiological saline, dispersed and filtered to remove large tissue fragments. Smears were prepared and stained with 1% Eosin Y. A total of 1000 sperm were counted /animal, scoring different types of sperm abnormalities. Scoring was performed under magnification (25×40) X with light microscope. In all Experiments negative (non-treated) and positive (Endoxan) control groups were run alongside the test material. Control groups treated orally with different soybean seeds extracts were also tested.
Sperm -Shape Abnormalities: The results show that the three tested doses of NiCl2 induced a statistically significant increase in the percentage of sperm abnormalities (P < 0.01) which show positive correlation with the dose Table (5). Various morphological sperm abnormalities in heads and tails were observed. Some heads may acquire unusual shape; others show a reduced or increased size. Coiled tail abnormality was also recorded.
Statistical Analysis: The significance of the difference between experimental and control data was calculated using t-test. The resulting data between groups (NiCl2treated and NiCl2+soybean) were compared by ANOVA, followed by Duncan’s multiple-range test.
Protective Role of Soybean: Figures 1 and 2 show the percentage of chromosomal aberrations in mouse bone-marrow and spermatocytes, comparing the
Table 1:Chromosomal aberrations induced in mouse bone marrow cells 24h after a single i.p. treatment with different doses of nickel chloride (NiCl2). Abnormal metaphases
Number of metaphases with different types of aberrations
375 metaphases examined in five mice per treatment.
R.T= Robertsonian translocation;
M.A. = metaphases with more than one type of aberration. ** Highly significant P < 0.01 level
and or break
Global J. Pharmacol., 8 (4): 625-634, 2014 Table 2: Percentage of metaphases with chromosomal aberrations induced in mouse bone-marrow cells after repeated i.p. treatment for 1,2 and 3 weeks with different doses of nickel chloride (NiCl2) and fed normal diet. Abnormal metaphases -------------------------------------------------------------Treatment and doses I- Control (non-treated)
Mean% ± S.E.
Mean% ± S.E.
4.5 ± 0.32
1.82 ± 0.32
13.86 ± 0.54**
2.93 ± 0.78
19.73 ± 0.54**
6.64 ± 0.60**
22.10 ± 0.90**
7.70 ± 0.78**
19.73 ± 0.51**
7.70 ± 0.79**
21.86 ± 0.98**
8.80 ± 0.99**
23.73 ± 0.51**
10.40 ± 0.50**
23.20 ± 0.60**
10.90 ± 0.63**
24.50 ± 0.99**
14.13 ± 0.90**
28.26 ± 0.65**
14.66 ± 0.85**
II- Nickel chloride 1 week treatment: 2.625 5.250 mg kg
10.50 2 weeks treatment: 2.625 5.250 mg kg
10.50 3 weeks treatment: 2.625 5.250 mg kg
** Significant at (0.01) level (t-test). Table 3: Chromosomal abnormalities induced in mouse spermatocytes 24h after a single i.p. treatment with different doses of Nickel chloride (NiCl2) Abnormal Metaphases
Number of metaphases with different types of aberrations
500 metaphases examined in five mice per treatment.
M.A. = metaphases with more than one type of aberration. ** = Highly significant P < 0.01 level.
Table 4: Percentage of metaphases with chromosomal abnormalities induced in mouse spermatocytes after repeated i.p. treatment for 1, 2 and 3 weeks with different doses of nickel chloride (NiCl2) and fed normal diet. Treatment and doses
I- Control (non-treated)
Examined metaphases No. 500
Abnormal metaphases No. 181
Abnormal metaphasesMean% ± S.E. 3.60 ± 0.51
6.20 ± 0.37**
9.60 ± 0.75**
11.20 ± 0.73**
9.00 ± 0.84**
13.20 ± 0.49**
14.00 ± 0.55**
9.40 ± 0.51**
15.20 ± 0.73**
17.00 ± 0.71**
II- Nickel chloride 1 week treatment 2.625 5.250 mg kg
10.50 2 weeks treatment: 2.625 5.250 mg kg
10.50 3 weeks treatment: 2.625 5.250 mg kg
10.50 ** Significant at (0.01) level (t-test).
Global J. Pharmacol., 8 (4): 625-634, 2014 Table 5: Morphological sperm abnormalities induced in male mouse after i.p. treatment with different doses of nickel chloride (NiCl2). Tail
Abnormal sperms -----------------------
Head abnormalities -------------------------------------------------------------------------------------------
abnormali - ties
Mean ± S.E
I. control (non treated )
2.46 ± 0.14
4.28 ± 0.31** 5.12 ± 0.23**
6.38 ± 0.20**
2.22 ± 0.36
2.32 ± 0.22 2.42 ± 0.24
3.20 ± 0.48
3.14 ± 0.13*
III. Mice fed
(+ ve control)
No of different types of sperm abnormalities -----------------------------------------------------------------------------------------
5000 sperm were counted five mice per treatment (1000 sperm per mice)
* Significant P < 0.05 level
** Highly significant P < 0.01 level.
Fig. 1: Percentage of chromosomal abnormalities (including gaps) induced in mouse bone-marrow cells after i.p. treatment with NiCl2 at 5.25 mg kg 1 bw , NiCl2+soybean seeds feeding and NiCl2 with "M.E.", "n-hx. F.", "Ch.F." or "M.F." of soybean seeds for 1 week (A), 2 weeks (B) and 3 weeks (C)
Fig. 2: Percentage of chromosomal abnormalities induced in mouse spermatocytes after i.p. treatment with NiCl2 at 5.25 mg kg 1 bw , NiCl2+soybean seeds feeding and NiCl2 with "M.E.", "n-hx. F.", "Ch.F." or "M.F." of soybean seeds for 1 week (A), 2 weeks (B) and 3 weeks (C). It is worth mentioning that, controls orally administered soybean extracts have the same results as control non-treated after cytological examination results obtained in mice treated with NiCl2 at the dose 5.25 mg Kg 1b.wt (1/8 LD50) and feed normal diet and that treated with NiCl2 and feed diet mixed with soybean seeds or orally treated with different soybean extracts (Methanol extract, n-hexane fraction, chloroform fraction or methanol fraction).
The results show that feeding mice with soybean seeds reduced the percentage of chromosomal aberrations in NiCl 2-treated groups in bone marrow and mouse spermatocytes. The results also revealed variable effects of soybean extracts and fractions. All of them reduced the percentage of 629
Global J. Pharmacol., 8 (4): 625-634, 2014
NiCl2-induced chromosomal aberrations. Methanol extract and methanol fraction gave the most effective protection. With respect to sperm abnormalities, Table( 5) shows that feeding NiCl -treated mice with soybean seeds
chromosomes. The absence of bivalents and the formation of unpaired chromosomes at the time of induction of the first anaphase, may lead to random segregation of homologous chromosomes i.e aneuploidy . The results show that X-Y univalent was more frequent. This type of abnormality has been discussed as an indicator of male sterility . Translocations in the form of Chain IV were observed. The maximum percentage reached 5% after repeated treatment with the highest tested dose which is relatively high. Translocations are one of the commonest forms of rearrangement in man. Fragments/breaks and gaps were also recorded in a low percentage. Morphological sperm abnormalities with dosedependent manner were observed in the present study. Both head and tail abnormalities were recorded. The head abnormalities most probably reflect a change in DNA content . Coiling of sperm tail mainly involves its orientation, which give an impression of a reduced sperm movement. Such limitation in sperm movement was reported to reduce fertility in both animals  and humans . It was reported that non-motile sperms lacked the dynein arms which is an ultra-structural abnormality of the microtubules in the tail motor complex . There is evidence that coiled-tail abnormality induced by other metals .The results of the present work are coincide well with the finding of other authors who demonstrated dose-dependent changes in sperm motility, count and abnormalities in mice treated with nickel chloride [33,34]. Oxidative stress mechanisms are speculated to play a significant role in nickel-induced genotoxicity . Nickel compounds induce genetic instability and chromosome instability which could result in multiple genetic alterations and are critically involved in carcinogenesis . Nickel II is also able to significantly enhance the genotoxicity of other mutagens and carcinogens  and interfere with DNA-repair mechanism and this may contribute to its genotoxicity . The results also demonstrated a significant decrease in the percentage of chromosomal aberrations and sperm abnormalities in all treated-mice feed soybean or orally administrated soybean extracts. Our results are in agreement with the previously reported data concerning the antimutagenic properties of soybean. Soybean reduced chromosomal damage and aberrations, DNAbreaks, point mutation and the effect on mitotic indices induced in mammalian cells by chemical mutagens e.g. the carbamate insecticide sevein ; the nitrosamine
(during 35 days until sacrificed) was effective in reducing different kinds of sperm abnormalities induced by NiCl 2. DISCUSSION The results of the present study showed that nickel chloride at the tested concentrations induced a significant increase in the percentage of chromosomal abnormalities in bone marrow and mouse spermatocytes after single and repeated i.p treatments, with a dose and time- response. The repeated dosing caused a higher percentage of chromosomal aberrations than the single-dose treatment which indicates a cumulative effect of NiCl2. Similar results have been reported after repeated treatments with other metals, e.g. lead acetate, potassium dichromate, cobalt chloride and beryllium chloride . Different types of chromosomal aberrations were observed in bone marrow cells after treatment with NiCl 2. Such results are in agreement with the previously published data demonstrated various types of aberrations included gaps, breaks and exchanges in FMA mouse mammary carcinoma cells, in cultured Chinese hamster ovary(CHO) cells, in mouse bone marrow and cultured spleen cells.The resulting data also confirm the findings of other authors who reported that inhalation exposure of Wister rats to metal aerosol derived from nickel refinery waste for a period of 4- months resulted in a significant increase in chromosomal aberrations in alveolar macrophages of Wister rats indicating its genotoxicity. Genetic effect of NiCl2 in somatic cells was also demonstrated by increase in the frequency of sister chromatid exchanges (SCE?s) and induction of micronuclei in peripheral blood and mouse bone marrow [14, 26]. Epidemiological studies also showed mutagenic risk in workers exposed to nickel compounds as evidenced by increasing in the frequency of chromosomal aberrations and SCE's and inhibition of DNA-repair synthesis . Nickel chloride induced chromosomal abnormalities in mouse spermatocytes which give information on transmissible genetic damage. It was observed that dissociated univalents dominated (autosomal and X-Y univalents). The process of chromosome pairing and the formation of bivalents during meiosis are necessary for proper genetic recombination and segregation of 630
Global J. Pharmacol., 8 (4): 625-634, 2014
precursors, dibutylamine and sodium nitrate  and acetoxyacetylaminoflurene . Fahmy et al  reported that, feeding mice with soybean diet reduced the genotoxicity of potassium dichromate as indicated by a significant reduction in the induced SCE's, chromosomal aberrations in bone marrow and spermatocytes and sperm abnormalities. Studies on the protective role of soybean showed that the anticarcinogenic/antimutagenic properties of soybean depend on its high nutritional value and to the presence of some active ingredients. Soybean is an excellent source of protein and amino acids. The protective role of amino acids for minimizing the incidence of chemically induced tumors was reported  Soybean is also a good source of ascorbic acid, nicotinic acid, vitamin A, thiamin, riboflavin and contain fibers .Vitamin C is especially important because it combats the effect of many chemical toxins including heavy metals and participating in DNA-repair processes . Soybean fibers (non starch polysaccharide) are regarded to have the property of eliminating mutagens by absorbing them and thus have no chance to reach the target cells . Protease inhibitors are another natural ingredients demonstrated in soybean. It was reported that the Bowman-Birk (BBI) which derived and isolated from soybean is the most potent one of the known protease inhibitors. It can contract the formation of oxygen radicals and play a major role in the prevention and or treatment of several diseases in addition to cancer, at very low concentrations [17, 44]. Phytic acid content also seems to be responsible for some of the antimutagenic properties of soybean. It is a highly charged antioxidant and has antitumor effects . Soybean contains non-nutrient bioactive phytochemicals that have health promising and disease preventing properties like isoflavones, saponins, coumestans and lignans. Concerning the protective effect of various soybean extracts, the results revealed that the methanol fraction and methanol extract gave the most effective protection and maintained all parameters studied at near the control values. Preliminary phytochemical studies on the prepared methanol fraction and extract revealed the presence of isoflavones glycosides and saponins as the main constituents. Anthony  reported that 90% of isoflavones could be extracted by alcohol. Isoflavones have been proposed to be the active component responsible for the beneficial effects of soybean foods and appear to work in conjunction with certain peptides or protein fractions from soybean to protect against many diseases and cancer . Isoflavones of soybean
possessed 4 to 5 folds antioxidant activity as ascorbic acid. Such antioxidant properties can be mediated by scavenging and/or suppressing the formation of reactive oxygen and/or nitrogen species and up- regulating antioxidant defense mechanism . The principle isoflavones in soybean are genistein and daidzein and their metabolities. These constituents seem to hold the most promise from the therapeutic stand point. Genistein and daidzein derived from soybean have antimutagenic/anticarcinogenic properties [50,51] and might exert their effect via hormonal regulation activity, controlling cell enzymes and inducing apoptosis [51,52]. Furthermore, genistein inhibits angiogenesis, the growth of blood vessels needed for tumors to enlarge . Saponins, the other major constituent detected in the prepared methanol fraction/extract were reported to have antimutagenic activity against mutagens induced DNAdamage and exhibit more potent activities than vitamin E and C [54, 55]. Soyasaponins appear to have anticancer properties by virtue of their antioxidant and antimutagenic properties . CONCLUSION In view of the results of the present work and the literature reviewed concerning the genotoxicity of nickel chloride, it can be concluded that the environmental contamination by this industrial metal may bring serious problems to human health including genetic hazards. The results also demonstrated the potential role of soybean and its components as natural protective antimutagenic agents and open the door for developing new drugs from this natural source. REFERENCES 1. 2. 3.
EHC, 1991. Environmental Health Criteria (Nickel) 108, Nickel World Health Organization, Geneva. Salnikow, K. and X. Li, 2004. Lippmann M. Effect of nickel and iron co-exposure on human lung cells. Toxicol. Appl. Pharmacol., 196: 258-65. Prasad, L., T.H. Khan, T. Jahangir and S. Sultana, 2007. Effects of luteolin on nickel - induced renal hyperproliferation and biotransformation parameters in Wister rats. Pharmaceutical Biology, 45: 116-123 Kang, G.S., P.A. Gillespie, A. Gunnison, A. Moreira, K-M. Tchou-Wong and L.C. Chen, 2011. Long-term inhalation exposure to nickel nanoparticles exacerbated atherosclerosis in susceptible mouse model. Environ Health Presp., 119: 176-181.
Global J. Pharmacol., 8 (4): 625-634, 2014
Ohshima, S., 2003. Induction of genetic instability and chromosomal instability by nickel sulfate in V79 Chinese hamster cells. Mutagenesis, 18: 133-137. Sunderman, F.W., S.K. Shen, J.M. Mitchell, P.R. Allpass and I. Damjanov, 1978. Embryo toxicity and fetal toxicity of nickel in rats. Toxicol. Appl. Pharmacol, 43: 381-390. Mathur, N., G. Pandey and G.C. Jain, 2010. Male reproductive toxicity of some selected metals : A review. J Biological Sciences, 10: 396-404. Ciub r, R., A. Cimpean and D. Iord chescu, 2006. Effects of nickel (II) on human polymorphonuclear leukocyte function in vitro. Revue Roumaine de Chimie, 51: 1169-1173. Coogan, T.P., D.M. Latta, E.T. Snow and M. Costa, 1989. Toxicity and carcinogenicity of nickel compounds. Crit Rev. Toxicol., 19: 341-384. Sunderman, F.W. Jr. 1989. Mechanisms of nickel carcinogenesis. Scand J. Work Environ Health, 15: 1-12. Ouyang, W., D. Zhang, J. Li, U.N. Verma, M. Costa and C. Huang, 2009. Soluble and insoluble nickel compounds exert a differential inhibitory effect on cell growth through 1KKá - dependent cyclin D1 down- regulation. J. Cell Physiol., 218: 205-214. Larramendy, M.L., N.C. Popescu and J.A. Dipaolo. 1981. Induction by inorganic metal salts of sister chromatid exchanges and chromosome aberrations in human and Syrian hamster cell strains. Environ Mutagen, 3: 597-606. Chakrabarti, S.K., C. Bai and K.S. Subramanian, 2001. DNA-protein crosslinks induced by nickel compounds in isolated rat lymphocytes: role of reactive oxygen species and specific amino acids. Toxicol. Appl. Pharm, 170: 153-65. Donya, S.M. and A.E. Ibrahim, 2006. Mutagenic responses of nickel chloride in somatic and germ cells of mice in vitro and in vivo studies. J. Egypt Soc. Toxicol, 30: 53 -59. Danadevi, K., R. Rozati, B. Saleha Banu and P. Grover, 2004. In vivo genotoxic effect of nickel chloride in mouse leukocytes using comet assay. Food Chem Toxicol, 42: 751-757. Tanaka, S., S. Koizumi, N. Makiuchi, Y. Aoyagi, E. Quivy, R. Mitamura, T. Kano, D. Wakita, K. Chamoto, H. Kitamura and T. Nishimura, 2011. The extract of Japanese soybean, Kurosengoku activities. The production of IL-12 and IFN-ã by DC or NK1-1 (+) cells in TLR4- and TLR2- dependent manner. Cell Immunol, 266: 135-42.
17. Palavalli, M.H., S.S. Natarajan, T.T. Wang and H.B. Krishnan, 2012. Imbibition of soybean seeds in warm water results in the release of copious amounts of Bowman- Birk protease inhibitor, a putative anticarcinogenic agent. J. Agric. Food Chem, 60: 3135-43. 18. Al-Ashaal, H.A., M.A. Fahmy, F.R. Melek, H.N. Aly and Z.M. Hasan, 2012. Effect of supplemented soybean (Glycine max.L ) diet and extracts on aluminum sulfate-induced genotoxicity. Toxicol Environ Chem, 94: 965-986. 19. Yosida, T.H. and K.Amano, 1965. Autosomal polymorphism in laboratory bred and wild Norway rats, Rattus norvegicus found in Misima. Chromosoma, 16: 658-667. 20. Evans, E.P., G. Breckon and C.E. Ford, 1964. An air drying method for meiotic preparations from mammalian testes. Cytogenetics. 3: 289-294. 21. Wyrobek, A.J. and W.R. Bruce, 1978. The induction of sperm-shape abnormalities in mice and humans, in: A Hollaender, FJ de serres (Eds), Chemical Mutagens: Principles and Methods for Their Detection, Plenum Press, New York, 5: 257-285. 22. Fahmy, M.A., N.H. Hassan, A.A. Farghaly and Hassan E.E. 2008. Studies on the genotoxic effect of beryllium chloride and the possible protective role of selenium/vitamins A, C and E. Mut. Res., 652: 103-11. 23. Nishimura, M. and M. Umeda, 1979. Induction of chromosomal aberrations in cultured mammalian cells by nickel compounds. Mut. Res., 68: 337-349. 24. Howard, W., B. Leonard, W. Moody and T.S. Kochhar, 1991. Induction of chromosome changes by metal compounds in cultured CHO cells. Toxicol. Lett., 56: 179-86. 25. Chorvatovicova, D. and Z. Kovacikova, 1992. Inhalation exposure of rats to metal aerosol. II. Study of mutagenic effect on alveolar macrophages. J .Appl. Toxicol., 12: 67-8. 26. Rabbani, S.I., K. Devi, S. Khanam and N. Zahra, 2006. Citral, a component of lemongrass oil inhibits the clastogenic effect of nickel chloride in mouse micronucleus test system. Pak J. Pharm Sci., 19: 108-13. 27. Perminova, I.N., N.I. Alekhina, T.A. Sinel'Shchikova, T.B. Osipova and G.D. Zasukhina, 1997. Formation of sister chromatid exchanges and reparative DNA synthesis in workers exposed to nickel compounds. Genetika, 33: 556-60. 632
Global J. Pharmacol., 8 (4): 625-634, 2014
28. Sosnowski, J., A. Lukaszewicz, L. Migalska, M. Wojnowska and Z.Polañski, 2011. Different levels of a lack of X-Y chromosome pairing in pachytene spermatocytes of red fox (Vulpes vulpes) and Chinese raccoon dog (Nyctereutes procyonoides procyonoides). Ann Anim Sci., 11: 71-81. 29. Burgoyne, P.S., S.K. Mahadevaiah, M.J. Sutcliffe and S.J. Palmer, 1992. Fertility in mice requires X-Y pairing and a Y chromosomal? spermiogenesis? gene mapping to the long arm. Cell, 71: 391-398. 30. Molnar, A., P. Sarlos, G. Fancsi, J. Ratky, S. Nagy and A.Kovacs, 2001. A sperm tail defect associated with infertility in a goat -case report. Acta Vet Hung., 49: 341-348. 31. Baccetti, B., T. Renieri, F. Rosati, M.G. Selmi and S. Casanova, 1977. Further observations on morphogenesis of the round headed human spermatozoa. Andrologia, 9: 255-264. 32. Pedersen, M. and H. Rebbe, 1975. Absence of arms in axoneme of immobile human spermatozoa. Biol Reprod. 12: 541-544. 33. Pandey, R. and S.P. Srivastava, 2000. Spermatotoxic effects of nickel in mice. Bull. Environ. Contam. Toxicol., 64: 161-167. 34. Doreswamy, K., B. Shrilatha, T. Rajeshkumar, Muralidhara, 2004. Nickel-induced oxidative stress in testis of mice : evidence of DNA damage and genotoxic Effects. J. Androl, 25: 996 -1003. 35. Deng, C.Z., M.P. Fons, J. Rosenblatt, R.A. EL-Zein, S.Z. Abdel-Rahman and T. Albrecht, 2006. Nickel potentiates the genotoxic effect of benzo [a] pyrene in Chinese hamster lung V79 cells. Environ Mol. Mutagen, 47: 150-61. 36. Carmona, E.R., A. Creus and R. Marcos, 2011 Genotoxic effects of two nickel compounds in somatic cells of Drosophilla melanogaster. Mut. Res., 718: 33-7. 37. El-Fiky, S.A., K.B. Abdel Aziz and S.A. Abdel Baset,. 1992. Protective role of soybean against chromosomal and biochemical effects of carbaryl (sevin) in male mice. J. Egypt Soc. Toxicol, 9: 15-23. 38. Tohamy, A.A., A.A. El-Ghor, N.Z. Moharram and M.M. El-Shazly,1996. Protective role of soybean feeding against the cytogenetical and histopathological effects of dibutylamine and sodium nitrate on bone- marrow and liver of mice. Mut. Res., 360: 155-163.
39. Plewa, M.J., E.D. Wagner, L. Kirchoff, K. Repetny, L.C. Adams and A.L. Rayburn, 1998. The use of single cell gel electrophoresis and flow cytometry to identify antimutagens from commercial soybean byproducts. Mut. Res., 402: 211-218. 40. Fahmy, M.A., H.M.Shoman and E.E. Hassan, 2002. The protective role of thiola and soybean seeds against the genotoxicity induced by potassium dichromate in mice. Mut. Res., 517: 1-12. 41. Gray, J.I. and J.R. Dugan, 1975. Inhibition of N-nitrosamine formation in model food systems. J. Food Sci., 40: 981. 42. Nwokolo, E., 1996. Soybean. IN : Food and Feed from Legumes and Oil seeds.E Nwokolo and J Smartt(eds) New York. pp: 90-101. 43. Rao, B.S., 1988. Dietary fiber in Indian diets and its nutritional significance. NFI Bull, 9: 1-5. 44. Kennedy, A.R., 1998. Chemopreventive agents: Protease inhibitors. Pharmacol Ther., 78: 167-209. 45. Vucenik, I., V.J. Tomazic, D. Fabian and A.M. Shamsuddin, 1992. Antitumor activity of phytic acid (inositol hexaphosphate) in murine transplanted and metastatic fibrosarcoma, a pilot study. Cancer Lett., 65: 9-13. 46. Anthony, M.S., T.B. Clarkson, C.L. JR Hughes, T.M. Morgan and G.L. Burke, 1996. Soybean isoflavones improve cardiovascular risk factors without affecting the reproductive system of peripubertal rhesus monkeys. J. Nutr., 126: 43-50. 47. Omoni, A.O. and R.E. Aluko, 2005. Soybean foods and their benefits: Potential mechanisms of action. Nutr. Rev., 63: 272-83. 48. Dai, J. and R.J. Mumper, 2010. Plant phenolics: Extraction, analysis and their antioxidant and anticancer properties. Molecules, 15: 7313-7352. 49. Kathi Head, N.D., 1998. Soy isoflavones and other constituents. Alternative Medicine Review, 2: 433-450. 50. Miyazawa, M., K. Sakano, S.I. Nakamura and H. Kosaka, 1999. Antimutagenic activity of isoflavones from soybean seeds.(Glycine max merrill). J. Agric. Food Chem, 47: 1346-1349. 51. Li, H.Q., Y. Luo and C.H. Qiao, 2012.The mechanisms of anticancer agents by genistein and synthetic derivatives of isoflavone. Mini Rev Med Chem, 12: 350-62. 52. Wang, T.T., N.Sathyamoorthy and J.M. Phang, 1996. Molecular effects of genistein on estrogen receptor mediated pathways. Carcinogenesis, 17: 271-275. 633
Global J. Pharmacol., 8 (4): 625-634, 2014
53. Zhou, J.R., E.T. Gugger, T. Tanaka, Y. Guo, G.L. Blackburn and S.K. Clinton, 1999.Soybean Phytochemicals inhibit the growth of transplantable human prostate carcinoma and tumor angiogenesis in mice. J Nutr., 129: 1628-1635 54. Berhow, M.A., E.D. Wagner, S.F. Vaughn and M.J. Plewa, 2000. Characterization and antimutagenic activity of soybean saponins. Mut. Res., 448: 11-22.
55. Jun, H.S., S.E. Kim and M.K. Sung, 2002. Protective effect of soybean saponins and major antioxidants against aflatoxin B1-induced mutagenicity and DNAadduct formation. J. Med Food, 5: 235-40. 56. Yang, X., C. Dong and G. Ren, 2011. Effect of soyasaponins-rich extract from soybean on acute alcohol-induced hepatotoxicity in mice. J. Agric. Food Chem, 59: 1138-44.