Ambreen Fatima

Evaluation of the Toxic Potential of Graphene Copper Nanocomposite (GCNC) in the Third Instar Larvae of Transgenic Drosophila melanogaster (hsp70-lacZ)Bg9

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)Bg9. 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).

Effect of Centella asiatica Leaf Extract on the Dietary Supplementation in Transgenic Drosophila Model of Parkinson's Disease

The role of Centella asiatica L. leaf extract was studied on the transgenic Drosophila model flies expressing normal human alpha synuclein (h-αS) in the neurons. The leaf extract was prepared in acetone and was subjected to GC-MS analysis. C. asiatica extract at final concentration of 0.25, 0.50, and 1.0 μL/mL was mixed with the diet and the flies were allowed feeding on it for 24 days. The effect of extract was studied on the climbing ability, activity pattern, lipid peroxidation, protein carbonyl content, glutathione content, and glutathione-S-transferase activity in the brains of transgenic Drosophila. The exposure of extract to PD model flies results in a significant delay in the loss of climbing ability and activity pattern and reduced the oxidative stress (P < 0.05) in the brains of PD flies as compared to untreated PD flies. The results suggest that C. asiatica leaf extract is potent in reducing the PD symptoms in transgenic Drosophila model of Parkinsons disease.

Protective effect of Geraniol on the transgenic Drosophila model of Parkinson’s disease

The role of Geraniol was studied on the transgenic Drosophila model flies expressing normal human alpha synuclein (h-αS) in the neurons. Geraniol at final concentration of 10, 20 and 40μM were mixed in the diet and the flies were allowed to feed on it for 24 days. The effect of geraniol was studied on the climbing ability, activity pattern, lipid peroxidation, protein carbonyl, glutathione, dopamine content, and glutathione-S-transferase activity in the brains of transgenic Drosophila. The exposure of PD model flies to 10, 20 and 40μM of geraniol results in a significant delay in the loss of climbing ability (p<0.05), improved activity pattern reduced the oxidative stress (p<0.05) in the brains of transgenic Drosophila as compared to unexposed PD model flies. The results suggest that geraniol is potent in reducing the PD symptoms in transgenic Drosophila model of Parkinsons disease.

Effect of bromocriptine alginate nanocomposite (BANC) on a transgenic Drosophila model of Parkinson's disease.

ABSTRACT The effect of bromocriptine, a dopamine agonist, administered in the form of bromocriptine alginate nanocomposite (BANC) was studied on Parkinsons disease (PD) model flies. The synthesized BANC was subject to characterization and, at a final concentration of 0.5, 1.0 and 1.5 µM, was mixed in diet. The PD flies were allowed to feed on it for 24 days. A significant dose-dependent delay in the loss of climbing activity and activity pattern was observed in PD flies exposed to 0.5, 1.0 and 1.5 µM BANC. The PD flies exposed to BANC also showed a significant reduction in lipid peroxidation and glutathione-S-transferase activity, and an increase in glutathione content. However, no gross morphological changes were observed in the brains of PD flies compared with controls. The results suggest that BANC is effective in reducing the PD symptoms in these transgenic flies. Summary: The results suggest that the bromocriptine alginate nanocomposite is potent in reducing the symptoms of Parkinsons disease in a transgenic fly model of the disease.

Toxic potential of copper-doped ZnO nanoparticles in Drosophila melanogaster (Oregon R)

Abstract Aims: In the present study, copper-doped ZnO nanoparticles (doped ZnO NPs Cu) were synthesized, characterized and evaluated for their possible toxic effects in Drosophila melanogaster (Oregon R). Methods and results: X-ray diffraction, Fourier transform infrared spectroscopy, scanning electron microscopy and energy dispersive X-ray spectrometry confirm the formation of doped ZnO NPs Cu. Doped ZnO NPs Cu (3%) were mixed in the diet at final concentrations of 1, 2, 4 and 8 µg/µl. The starved male flies were allowed to feed on it for 4 days. After completion of the desired duration, climbing ability, activity pattern, activity of acetylcholinesterase (AChE), glutathione (GSH), glutathione-S-transferase (GST), lipid peroxidation (LPO), total protein content and caspases were studied. SDS–PAGE was also performed for whole fly homogenate of control as well as treated flies. No loss in the climbing and activity pattern was observed at the selected doses of doped ZnO NPs Cu. No significant change in the levels of AChE, GSH, GST, LPO, caspase 9/3 and total protein content was observed. The brain sections showed no gross changes in the structure and SDS–PAGE patterns also revealed no change in the protein expression. Conclusions: The results suggest that doped ZnO NPs Cu are non-toxic at 1, 2, 4 and 8 µg/µl of concentration in D. melanogaster.

Effect of Tangeritin Against Cyclophosphamide-Induced Toxicity in the Larvae of Transgenic Drosophila melanogaster (hsp70-lac Z) Bg9

ABSTRACT Tangeritin is a pentamethoxy flavone found in tangerine and other citrus peels. The present study was conducted to study the effect of various doses of tangeritin on the toxic effects induced by cyclophosphamide (CP) in the third instar larvae of transgenic Drosophila melanogaster (hsp70-lacZ)Bg9. Commercially available tangeritin and CP were used. CP was used alone at 20 µM of food concentration and with 20, 40, and 80 µM of tangeritin. The exposure of the third instar larvae to 20µM of cyclophosphamide alone results in significant hsp 70 expression along with tissue, DNA, and oxidative damage. The exposure of the third instar larvae to 20 µM of CP along with 20, 40, and 80 µM of tangeritin results in a dose-dependent decrease not only in hsp 70 expression but also in tissue, DNA, and oxidative damage. The results of the present study suggest that tangeritin has a protective role against the toxic effects of cyclophosphamide in the third instar larvae of transgenic Drosophila melanogaster (hsp 70-lac Z)Bg9.

Detection of aneugenicity and clastogenicity in buccal epithelial cells of pan masala and gutkha users by pan-centromeric FISH analysis

Chewing of betel quid, smoking and alcohol consumption are all associated with higher incidences of oral cancer. Genetic damage can be detected by fluorescent in situ hybridization (FISH) using human centromeric probes. In the present study FISH was performed on buccal epithelial cells of pan masala and gutkha chewers alone with and without additional tobacco smoking and/or alcohol consumption. The study comprised of 1500 male individuals. The present study found the highest frequency of micronuclei without a centromeric region (MN(-)) among gutkha users who also smoked and drank (P < 0.05). A significant increase in cells having micronuclei with a centromeric region (MN(+)) was observed among pan masala users who also smoked (P < 0.05). The study reveals that the clastogenic effects of pan masala/gutkha increase with smoking and alcohol consumption, but aneugenic effects were also observed among the pan masala chewers who smoked.

Evaluation of the toxic potential of calcium carbide in the third instar larvae of transgenic Drosophila melanogaster (hsp70-lacZ)Bg9

In the present study the toxic potential of calcium carbide (CaC2) was studied on the third instar larvae of transgenic Drosophila melanogaster (hsp70-lacZ)Bg(9). The third instar larvae were exposed to 2, 4, 8, 16 and 32×10(-3)g/ml of CaC2 in diet for 24h. The results reveal that the dose 2×10(-3)g/ml was not toxic but the remaining doses showed a dose dependent significant increase in the hsp70 expression, β-galactosidase activity, tissue damage, oxidative stress markers (lipid peroxidation and protein carbonyl content), glutathione-S-transferase activity, expression of Caspase 3 and 9, apoptotic index and DNA damage (midgut cells). A significant reduction as compared to control group in total protein, glutathione content and acetylcholinesterase activity was also observed. The Inductively Coupled Plasma Atomic Emission Spectroscopy analysis (ICPAES) reveals the presence of copper, iron, sodium, aluminium, manganese, calcium, nickel and mercury. The toxic effects of CaC2 in the present study may be attributed to the impurities present in it.

Effect of cabergoline alginate nanocomposite on the transgenic Drosophila melanogaster model of Parkinson’s disease

Abstract Parkinson’s disease (PD) is a common neurodegenerative disorder characterized by the loss of dopaminergic neurons in the substantia nigra of the midbrain. Dopamine agonists help the patients with PD by reversing the dopamine depletion and related motor deficits. In the present work, cabergoline, a potent ergot dopamine agonist, was given in the form of cabergoline alginate nanocomposite (CANC) to the PD model flies to study its effects on climbing ability, activity pattern, life span, lipid peroxidation, glutathione (GSH) content, glutathione-S-transferase (GST) activity, dopamine content, protein carbonyl content, mean gray-scale values, and caspase-3 and caspase-9 activities. Cabergoline alginate nanocomposite was synthesized by adding the cabergoline solution in the warm aqueous solution of sodium alginate; The synthesized CANC was characterized using fourier transform (FTIR) infrared spectroscopy, transmission electron microscopy (TEM), and UV–Visible spectroscopic techniques. The synthesized CANC having the final doses of 1, 2, and 3 μM was supplemented with diet and the flies were allowed to feed on it for 24 days. Cabergoline alginate nanocomposite significantly increases climbing ability, reduces lipid peroxidation, GST activity, protein carbonyl content, caspase 3/9 activity, mean gray-scale values, and increases the GSH as well as dopamine content in a dose-dependent manner. The results of this study suggest that CANC is potent in delaying and reducing the symptoms of PD.

Models of Parkinson’s Disease with Special Emphasis on Drosophila melanogaster

BACKGROUND & OBJECTIVE Parkinsons disease is the second most common neurodegenerative disorder affecting more than 1% of the population averaged 60 years of age. The majority of PD cases are sporadic and are probably caused by a combination of risk factors but 5-10% of the PD cases are familial. Due to the high degree of the gene, conservation in humans, mice and insects using an animal model system is a valuable approach to further elucidate the roles of the genes in PD. CONCLUSION The present review highlights the models used to study PD symptoms with special emphasis on Drosophila.