[show abstract][hide abstract] ABSTRACT: Oral submucosus fibrosis (OSMF) is a collagen-related disorder seen in habitual betel quids and smokers. This is a high risk precancerous condition in which the connective tissue fibers of the lamina propria and deeper parts of the mucosa becomes stiff with restricted mouth opening. Patients with severe cases have symptoms like difficulties in chewing, swallowing and speaking. In the present study 25 individuals were gutkha chewers and 25 were OSMF patients (chewing gutkha along with smoking) and 25 individuals were taken as controls. A significant increase in the frequency of micronuclei was observed in OSMF patients (34.4 ± 1.79) as compared to gutkha chewers (14.4 ± 0.73) and controls (4.36 ± 0.27). The number of micronucleated cells in OSMF, gutkha chewers and control groups were 19.84 ± 0.69, 12.6 ± 0.51 and 4.20 ± 0.27, respectively and are significantly different at p < 0.05. Acridine orange is used due its fluorescence nature and easier visibility of the micronucleus present in the buccal epithelial cells. It is concluded that chewing gutkha along with smoking is more dangerous for human health as it hastens the incidence of OSMF.
Egyptian Journal of Medical Human Genetics 04/2013; 14(2):189–193.
[show abstract][hide abstract] ABSTRACT: The role of Eucalyptus citriodora L. leaf extract was studied on the transgenic Drosophila model of flies expressing normal human alpha synuclein (h-αS) in the neurons. These flies exhibit locomotor dysfunction as the age progresses. The leaf extract was prepared in acetone and was subjected to GC-MS analysis. The GC-MS analysis revealed the presence of 9 major compounds. E. citriodora extract at final concentration of 0.25, 0.50 and 1.0μl/ml was supplemented with the diet and the flies were allowed to feed for 21 days. The effect of extract was studied on the climbing ability and the oxidative stress on the PD model Drosophila expressing normal human alpha synuclein (h-αS) in the neurons. The supplementation of 0.25, 0.50 and 1.0μl/ml of E. citriodora extract showed a dose dependent significant delay in the loss of climbing ability and reduction in the oxidative stress in the brain of PD model flies. The results also support the utility of this model in studying PD symptoms.
Food and chemical toxicology: an international journal published for the British Industrial Biological Research Association 01/2013; · 2.99 Impact Factor
[show abstract][hide abstract] ABSTRACT: In the present study the comet assay was performed in buccal epithelial cells to evaluate DNA damage among pan masala or gutkha chewers and smokers. The assay is a rapid, suitable and sensitive method for detecting various forms of DNA damage at individual cell level. The study comprises 300 individuals of which 50 individuals were gutkha chewers along with smoking, 50 indi-viduals were pan masala chewers along with smoking, 50 individuals were gutkha chewers, 50 indi-viduals were pan masala chewers, 50 individuals were smokers and 50 individuals were non-users (control) or not having any addiction. Comet tail length was observed to measure the extent of DNA damage. In all groups a significant increase in the tail length was observed as compared to the non-users (control). The highest tail length was observed among gutkha chewers along with smoking (36.9 ± 3.60). The results of the present study suggest that the panmasala and gutkha are genotoxic agents and induce DNA damage. Ó 2013 Production and hosting by Elsevier B.V. on behalf of Ain Shams University.
[show abstract][hide abstract] ABSTRACT: Graphene, a two-dimensional carbon sheet with single-atom thickness, have attracted the scientific world for its potential applications in various field including the biomedical areas. In the present study the graphene copper nanocomposite (GCNC) was synthesized, characterized and evaluated for its toxic potential on third instar larvae of transgenic Drosophila melanogaster (hsp70-lacZ)Bg(9) . The synthesized GCNC was analyzed by X-ray diffraction (XRD), scanning/transmission electron microscopy (SEM/TEM), atomic force microscopy (AFM), and fourier transform infrared spectroscopy (FTIR). The GCNC in 0.1% DMSO was sonicated for 10 min and the final concentration of 0.033, 0.099, 0.199 and 3.996 µg/µl of diet were established. The third instar larvae were allowed to feed on it separately for 24 and 48 hrs. The hsp70 expression was measured by O-nitrophenyl-β-D-galactopyranoside assay, tissue damage by trypan blue exclusion test and β-galactosidase activity was monitored by in situ histochemical β-galactosidase staining. Oxidative stress was monitored by performing lipid peroxidation assay and total protein estimation. Ethidium bromide/acridine orange staining was performed on midgut cells for apoptotic index and the comet assay was performed for the DNA damage. The results of the present study showed that the exposure of 0.199 and 3.996 µg/µl of GCNC were toxic for 24 hr of exposure and for 48 hr of exposure: 0.099, 0.199 and 3.996 µg/µl of GCNC was toxic. The dose of 0.033 µg/µl of GCNC showed no toxic effects on its exposure to the third instar larvae for 24 hr as well as 48 hrs. This dose can be considered as No Observed Adverse Effect Level (NOAEL).
PLoS ONE 01/2013; 8(12):e80944. · 3.73 Impact Factor
[show abstract][hide abstract] ABSTRACT: Synthetic progestins and estrogens have been reported to be toxic in various experimental models. Their prolonged use has been reported to induce cancer in humans. In the present study the effects of oral contraceptives were studied among users using chromosomal aberrations, sister chromatid exchanges and DNA damage as a parameter, in cultured human peripheral blood lymphocytes. The study was performed on 25 women (users) and 25 age match controls. No significant difference was observed in chromosomal aberrations and DNA damage. A significant increase was observed in sister chromatid exchanges (SCEs) Cell among users. The results obtained and the risk of oral contraceptives (OCs) genotoxicity have been discussed.
Egyptian Journal of Medical Human Genetics 10/2012; 13(3):301–305.
[show abstract][hide abstract] ABSTRACT: All living organisms respond to various physical or chemical stressors by the induction of heat shock protein (HSP). The present study was performed on transgenic Drosophila melanogaster (hsp70-lacZ) Bg(9) in which the transformation vector is inserted with a P-element, the line contains wild-type hsp70 sequence up to the lacZ fusion point. The effect of L-ascorbic acid on the hsp70 expression and tissue damage was studied at the doses of 1, 2, 4, and 8 × 10(-4) g/ml in the third instar larvae of transgenic D. melanogaster (hsp70-lacZ) Bg(9). The larvae were exposed to different doses of L-ascorbic acid for 24 and 48 hours. A dose-dependent significant increase in the hsp70 expression was observed at 2, 4, and 8 × 10(-4) g/ml of L-ascorbic acid for both 24 and 48 hours. The tissue damage was observed only in the 48 hours of exposure and mostly only in the salivary glands of the third instar larvae of transgenic D. melanogaster (hsp70-lacZ) Bg(9). The present study also validates and supports the use of transgenic D. melanogaster (hsp70-lacZ) Bg(9) for the toxicological evaluations.
[show abstract][hide abstract] ABSTRACT: ABSTRACT In the present study, the effect of l-ascorbic acid (AA) was studied on the climbing ability of the Parkinson's disease (PD) model Drosophila expressing normal human alpha synuclein (h-αs) in the neurons. These flies show locomotor dysfunction as the age progresses. AA at final concentration of 11.35 × 10(-5) M, 22.71 × 10(-5) M, 45.42 × 10(-5) M, and 68.13 × 10(-5) M was added to the diet, and the flies were allowed to feed for 21 days. AA at 11.35 × 10(-5) M did not show any significant delay in the loss of climbing ability of PD model flies. However, AA at 22.71 × 10(-5) M, 45.42 × 10(-5) M, and 68.13 × 10(-5) M showed a dose dependent significant (p < .05) delay in the loss of climbing ability of PD model flies as compared to the untreated PD flies. The total protein concentration in brain homogenate was measured in treated as well as control groups after 21 days, no significant difference was obtained between treated as well as control (PD flies and l-dopa) groups. The results suggest that AA is potent in delaying the climbing disability of the PD model flies expressing h-αs in the neurons.
The International journal of neuroscience 07/2012; · 0.86 Impact Factor
[show abstract][hide abstract] ABSTRACT: A method using 1-methyl-2-phenylindole was developed for the estimation of lipid peroxidation in third instar larvae of transgenic Drosophila melanogaster (hsp70-lacZ)Bg (9). The method is specific for the estimation of malonaldehyde.
The larvae were exposed to 0.0025, 0.025, 0.050, and 0.100 μl/ml of cyclophosphamide for 24 and 48 h. The homogenate was prepared of the larvae tissue explant and the absorbance was noted at 586 nm.
A significant dose-dependent increase in the mean absorbance values was observed for both 24 and 48 h of exposure as compared to the untreated group.
On the basis of results obtained, it is suggested that the present method is more precise, accurate, and robust for the estimation of lipid peroxidation in the third instar larvae of transgenic D. melanogaster (hsp70-lacZ)Bg (9).
[show abstract][hide abstract] ABSTRACT: Parkinson's disease (PD) is characterized by the progressive loss of dopaminergic neurons and the aggression of alpha Synuclein (αS) in the brain. Drosophila mutants and transgenes have provided a platform to understand the mechanistic insight associated with the degenerative diseases. A number of polyphenols have been reported to inhibit the αS aggregation resulting in the possible prevention of PD. The involvement of free radicals in mediating the neuronal death in PD has also been implicated. In the present study, the effect of Nordihydroguaiaretic acid (NDGA) was studied on the climbing ability of the PD model Drosophila expressing normal human alpha synuclein (h-αS) in the neurons. These flies exhibit locomotor dysfunction as the age progresses. NDGA at final concentration of 0.01, 0.1, 0.5, and 1μl/ml was supplemented with the diet and the flies were allowed to feed for the 24 days. NDGA at 0.01 μl/ml did not showed any significant delay in the loss of climbing ability of PD model flies. However, NDGA doses at 0.1, 0.5, and 1.0 μl/ml showed a dose dependent significant (p < .05) delay in the loss of climbing ability of PD model flies as compared to the untreated PD flies. The results suggest that the NDGA is potent in delaying the climbing disability of PD model flies and also supports the utility of this model in studying PD symptoms.
[show abstract][hide abstract] ABSTRACT: Gutkha is a cheap and convenient betel quid substitute, which is popular among all age groups. Various studies reveal its carcinogenic nature that leads to oral submucosus fibrosis and increases the chances of oral cancer. The micronucleus (MN) assay in exfoliated mucosal cells is a useful method for observing genetic damage in humans.
To observe the genotoxic effect of gutkha on human buccal epithelial cells.
The MN assay was performed to assess the frequency of MN in human buccal epithelial cells. The study comprises 60 individuals of which 30 individuals were gutkha chewers and another 30 were nonusers (control). The MN frequency was scored to estimate the genotoxic damage.
In gutkha users, the frequency of MN was highly significant (17.4 ± 0.944) as compared with nonusers (control) groups (4.53 ± 0.331) (P < 0.001).
The MN assay in human buccal epithelial cells is a useful and minimally invasive method for monitoring genetic damage in humans. Asignificantly higher frequency of micronucleated cells are found among gutkha users.
[show abstract][hide abstract] ABSTRACT: Malondialdehyde (MDA) is used for the estimation of damage by reactive oxygen species. MDA is a major reactive aldehyde resulting from the peroxidation of biological membranes. The most common method used to assess MDA production is the thiobarbituric acid (TBARS) assay. However, the value of this method is curbed by low specificity and has been criticized for its use in human studies. In the present study we have used an alternative method for the estimation of MDA production i.e. reaction of MDA with a chromogenic agent 1-methyl-2-phenylindole at 45°C. The paper describes the method of preparing standards for the estimation of MDA (lipid peroxidation) after the treatment with an oxidative stress inducing agent hydrogen peroxide (H(2)O(2)). In the present study, the treatments of 1, 5, 10, 20, 50, 100, 150 and 200 μM of H(2)O(2) induced significant increase in lipid peroxidation as compared to the untreated ones. The results suggest that the present method can be used to measure the lipid peroxidation in cultured human peripheral blood lymphocytes and is specific for MDA estimation.
[show abstract][hide abstract] ABSTRACT: Abstract In the present study the effect of 0.1, 0.2, 0.4, 0.8, and 1.0 µL/mL of the steroid K-canrenoate was evaluated in the third instar larvae of transgenic Drosophila melanogaster (hsp70-lacZ) Bg(9) for 6, 24, and 48 hours of duration. The treatment of 0.1, 0.2, and 0.4 µL/mL of K-canrenoate did not induce the activity of hsp70 significantly compared to the control. The treatments of 0.8 and 1.0 µL/mL of K-canrenoate not only caused tissue damage but also induced a significant increase in the expression of hsp70 for the different durations of exposure. The results of the present study suggest that the K-canrenoate at 0.8 and 1.0 µL/mL is cytotoxic and caused tissue damage in the third instar larvae of transgenic D. melanogaster (hsp70-lacZ) Bg(9).
Journal of Insect Science 01/2012; 12:92. · 0.88 Impact Factor
[show abstract][hide abstract] ABSTRACT: Methyl methanesulfonate (MMS) is an anti-carcinogenic drug and its toxicity has been reported in various experimental models. The hsp70s are a family of ubiquitously expressed heat shock proteins. In the recent years, hsp70 has been considered to be one of the candidate genes for predicting cytotoxicity against environmental chemicals. Nowadays emphasis is given to the use of alternatives to mammals in testing, research and education. The European Centre for the Validation of Alternative Methods (EVCAM) has recommended the use of Drosophila as an alternative model for scientific studies. Almost all living organisms possess proteins with a similar structure to that of hsp70s. In the present study, the toxicity of MMS was evaluated by quantifying hsp70 expression and tissue damage in the third instar larvae of transgenic Drosophila melanogaster (hsp70-lacZ) Bg(9), at different doses and hours of exposure. We studied the effect of 0.25, 0.50, 0.75 and 1.0 µl/ml of MMS at 2, 4, 24 and 48 hours of exposure on hsp70 expression by using the soluble O-nitrophenyl-β-D-galactopyranoside (ONPG) assay and on establishing the tissue damage by the Trypan blue exclusion assay in the third instar larvae of transgenic Drosophila melanogaster (hsp70-lacZ) Bg(9). A dose-dependent increase in the expression of hsp70 was observed at 0.25, 0.50, and 0.75 µl/ml of MMS compared to the control. At the highest dose, i.e. 1.0 µl/ml of MMS, the activity of hsp70 was decreased due to tissue damage.
[show abstract][hide abstract] ABSTRACT: Antigenotoxic activity of allicin, one of the sulphur compounds of garlic (Allium sativum) which possesses antioxidant and thiol disulphide exchange activity, was studied against estradiol-17beta-induced genotoxic damage using chromosomal aberrations (CAs) and sister chromatid exchanges (SCEs) as parameters. Approximately 10, 20 and 40 microM of estradiol-17beta was tested for its genotoxic effect in the presence of metabolic activation and was found to be genotoxic at 20 and 40 microM. Approximately 20 microM of estradiol-17beta was treated along with 5, 10 and 15 microM of allicin, separately, in the presence of metabolic activation. Similar treatments were given with 40 microM of estradiol-17beta. Treatments along with allicin result in the reduction of CAs and SCEs, suggesting its anti-genotoxic activity in human lymphocytes in vitro against estradiol-17beta-induced genotoxic damage.
Natural product research 07/2010; 24(12):1087-94. · 1.01 Impact Factor
[show abstract][hide abstract] ABSTRACT: In the present study the antigenotoxic effect of apigenin was studied against a genotoxic dose of ethinylestradiol using the damage parameters of chromosomal aberrations (CAs), sister chromatid exchanges (SCEs) and cell cycle kinetics (CCK). Human peripheral blood lymphocytes were cultured and treated with 10 microM of ethinylestradiol along with doses of 5, 10, 15 and 20 microM of apigenin. A clear decrease in the genotoxic damage induced by ethinylestradiol was observed with increasing doses of apigenin, suggesting a protective role for apigenin during ethinylestradiol therapy.
[show abstract][hide abstract] ABSTRACT: Mitomycin C (MMC) is an antineoplastic agent used to fight a number of different cancers including cancer of the stomach, colon, rectum, pancreas, breast, lung, uterus, cervix, bladder, head, neck, eye and oesophagus. It is a potent DNA cross-linker. The prolonged use of the drug may result in permanent bone marrow damage and other various types of secondary tumors in normal cells. The toxic effect of anticancerous drugs may be reduced if supplemented with natural antioxidants/plant products. With this view, the effect of 5, 10 and 15 microM of curcumin was studied against the genotoxic doses of MMC, i.e. 10 and 20 microM, in cultured human lymphocytes using cell viability, lipid peroxidation and DNA damage quantification as parameters. The treatment of curcumin with MMC results in a significant dose-dependent increase in cell viability and decrease in lipid peroxidation and DNA damage suggesting a protective role of curcumin against the anticancerous drug mitomycin C.
Experimental and toxicologic pathology: official journal of the Gesellschaft fur Toxikologische Pathologie 08/2009; 62(5):503-8. · 1.43 Impact Factor
[show abstract][hide abstract] ABSTRACT: Hydrogen peroxide is one of the reactive oxygen species for cellular injury. It is overproduced during oxidative stress and is known to damage proteins, nucleic acids and cell membranes. The present study was aimed to study the protective effect of ascorbic acid against the toxic doses of hydrogen peroxide using lipid peroxidation and cytokinesis blocked micronucleus assay. Hydrogen peroxide was studied at 50, 100 and 200μM and was found to increase a dose dependent increase in lipid peroxidation and micronuclei frequency. The treatment of 100 and 200μM of hydrogen peroxide separately along with 20, 40 and 80μM of ascorbic acid showed a dose dependent decrease in the lipid peroxidation and micronuclei frequency. The results suggest a protective effect of ascorbic acid against the hydrogen peroxide induced oxidative damage in cultured human peripheral blood lymphocytes.
Indian Journal of Clinical Biochemistry 07/2009; 24(3):294-300.
[show abstract][hide abstract] ABSTRACT: A sample of 312 individuals belonging to Sheikh (80), Pathan (54), Ansari (82), Syed (33), Saifi (33) and Hindu Bania (30) populations were surveyed for four protein and three enzyme loci comprising 12 alleles. The markers used were protein (transferrin, haptoglobin, haemoglobin) and enzyme (lactate dehydrogenase, phosphogluconate dehydrogenase, adenylate kinase). Except caeruloplasmin, variants were found among all loci, though no rare variant appears. The populations show genetic equilibrium for all of the loci. Our gene frequencies show some difference from earlier studies for Muslims in general, there being no biradari wise study among Sunni Muslims earlier done anywhere in India or abroad. Heterozygosity ranged from 0.0123 to 0.1994 (Sheikh), 0.0182 to 0.2046 (Pathan), 0.0239 to 0.1844 (Ansari), 0.0587 to 0.3966 (Syed), 0 to 0.3909 (Bania); the average DST and GST values for the seven marker loci were 0.001032 and 0.00879, respectively. The results are discussed. The gene frequency study shows closer relationship of Ansaris and Saifis with native Hindu Banias, and may reflect on their probable conversion in the not remote past.
[show abstract][hide abstract] ABSTRACT: Tea (Camellia sinensis) is one of the most popular beverages consumed worldwide as an infusion of leaves and isvalued for its medicinal properties. Tea is a rich source of polyphenols called flavonoids, effective antioxidants foundthroughout the plant kingdom. The slight astringent, bitter taste of green tea is attributed to polyphenols. A group offlavonoids in green tea are known as catechins, which are quickly absorbed into the body and are thought tocontribute to some of the potential health benefits of tea. The fresh tea leaves contain four major catechins ascolourless water soluble compounds. epicatechin (EC), epicatechin gallate (ECG), epigallocatechin (EGC) andepigallocatechin gallate (EGCG). Epidemiologic observations and laboratory studies have indicated that teapolyphenols act as antioxidants in vitro by scavenging reactive oxygen and nitrogen species and chelating redoxactivetransition metal ions and hence tea may reduce the risk of a variety of illnesses, including cancer and coronaryheart disease. In this study we seen the antigenotoxic effect of green tea extract against genotoxic damage inducedby two anabolic steroids Trenbolone and Methyltestosterone in cultured human lymphocytes, both in absence andpresence of metabolic activation. The results prove the antigenotoxic potential of green tea extract. Because theepidemiologic studies and research findings in laboratory animals have shown the antigenotoxic potential of teapolyphenols, the usefulness of tea polyphenols for various human diseases like cancer and coronary heart diseaseetc should be evaluated in clinical trials.