Effect of donepezil hydrochloride on the transgenic Drosophila expressing human Aß-42.

CONCLUSION: The results suggest that donepezil hydrochloride is potent enough to reduce the AD symptoms being mimicked in transgenic flies.

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

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.

Effect of capsaicin on the oxidative stress and dopamine content in the transgenic Drosophila model of Parkinson's disease.

In the present study the effect of capsaicin was studied on PD model flies expressing human alpha synuclein. First the potential of scavenging superoxide anion and free radicals by capsaicin at doses of 20, 40, 80 and 100 µM was estimated. The PD flies were allowed to feed separately on the diet containg 20, 40, 80 and 100 µM of capsaicin, respectively, for 24 days. After 24 days of exposure, fly head homogenate was prepared from each group and was used to estimate glutathione (GSH), protein carbonyl (PC), dopamine content, lipid peroxidation (LPO), glutathione-S-transferase (GST) and monoamine oxidase (MAO) activity. A dose dependent significant increase in the potential of scavenging superoxide anions and free radicals by capsaicin was observed for the doses of 20, 40, 80 and 100 µM. The exposure of capsaicin not only significantly increased the GSH (max. by 1.37-fold), and dopamine (max. by 1.56-fold) content but also reduced LPO (max. by 1.8-fold), GST (max. by 1.26-fold), MAO activities (max. by 1.60-fold) and PC content (max. by 1.95-fold), compared to unexposed PD flies (p < 0.05). The results suggest the protective role of capsaicin against the PD symptoms.

Effect of Oral Contraceptive Pills on the Blood Serum Enzymes and DNA Damage in Lymphocytes Among Users.

The continuous use of synthetic hormones as contraceptive pill or hormonal replacement therapy among women is increasing day by day. The widespread use of different formulations as oral contraceptives by women throughout their reproductive cycle has given rise to a serious concern for studying the effects of oral contraceptives on enzymatic profile and DNA damage in peripheral blood lymphocytes among users. The present study was carried out on women taking oral contraceptives. The study was based on the questionnaire having the information of reproductive history, fasting, age, health, nature of menstrual cycle, bleeding and other disease. The profile of the blood serum enzymes i.e. alkaline phosphatase (ALP), gamma glutamyl transferase (GGT), lactate dehydrogenase (LDH), aminotransferases (SGOT and SGPT), serum proteins (albumin and globulin) and DNA damage in lymphocytes was studied among users and non-users. The results of the present study suggest that OCs not only effects enzymatic activity but also results in DNA damage that may vary with the duration of using oral contraceptives. A significant increase in LDH, GGT, SGPT, SGOT, globulin and decrease in ALP as well as albumin was found among users as compared to non-users. The observed DNA damage was more in users as compared to non-users. Hormonal contraceptives seem to exert DNA damage and also have significant effects on blood serum enzymes.

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.

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

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 L-ascorbic Acid on the climbing ability and protein levels in the brain of Drosophila model of Parkinson's disease.

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.

Effect of L-ascorbic Acid on the hsp70 Expression and Tissue Damage in the Third Instar Larvae of Transgenic Drosophila melanogaster (hsp70-lacZ) Bg(9).

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.

Comparative study of rivastigmine and galantamine on the transgenic Drosophila model of Alzheimer's disease.

Alzheimer's Disease (AD) is characterized as a progressive neurodegenerative disease most commonly associated with memory deficits and cognitive decline. The formation of amyloid plaques and neurofibrillary tangles are important pathological markers of AD. The accumulation of amyloid plaques and neurofibrillary tangles leads to the loss of neurons including the cholinergic neurons thus decreasing the levels of acetylcholine (a neurotransmitter). To reduce the AD symptoms cholinesterase inhibitors are widely used to decrease the hydrolysis of acetylcholine released from presynaptic neurons. In the present study we have studied the effect of rivastigmine and galantamine (commonly used cholinesterase inhibitors) on the transgenic Drosophila model of AD expressing human Aß-42 in the neurons. The effect of similar doses of rivastigmine and galantamine (i.e. 0.1,1 and 10 â€‹mM) was studied on the climbing ability, lifespan, oxidative stress markers, caspase 9 and 3, acetylcholinesterase activity and on the formation of Aß-42 aggregates. The results suggest that the rivastigmine is more potent in reducing the oxidative stress and improving climbing ability of AD flies. Both the drugs were found to be effective in increasing the lifespan of AD flies. Galantamine was found to be a more potent inhibitor of acetylcholinesterase compared to rivastigmine. Galantamine prevents the formation of Aß-42 aggregates more effectively compared to rivastigmine.

Drosophila melanogaster a Versatile Model of Parkinson's Disease.

Parkinson's Disease (PD) is one of the most prevalent, recurrent and life-threatening neurodegenerative diseases. However, the precise mechanism underlying this disease is not yet clearly understood. For understanding the pathogenesis of PD, it is essential to identify the symptoms along with the novel biological markers and to develop strategies that could lead towards the development of effective therapy. PD is associated with Lewy bodies (LBs) formation and the loss of dopaminergic neurons in the substantia nigra pars compacta of mid brain region. For the improvement in treatment strategies, as well as understanding the pathophysiology of the PD in a number of animal models have been introduced that can recapitulate the pathophysiology, motor and nonmotor symptoms of PD. In contrast to mammalian models like rodents, mice and monkey, Drosophila is easy to handle as well as its maintenance cost is low. Due to the anatomical differences in the brain and other major organs of human and fly, the issues of standardizing the methods or experiments to analyze behavioral aspects (walking, writhing, eating and sleeping) are difficult in flies. The present review highlights the studies carried out for PD since 2000, using Drosophila melanogaster.

Nanotechnology: Its Application in Treating Neurodegenerative Diseases.

Neurodegenerative diseases such as Alzheimer's, Parkinson's and Huntington disease have serious concern due to its effect on the quality of life of affected persons. They have some limitations at diagnostic as well as the treatment level. Introducing nanotechnology, for the treatment of these diseases may contribute significantly to solve the problem. There are several treatment strategies for the neurodegenerative diseases, but most of them fail to cross the Blood-Brain Barrier (BBB). The present review highlights the application of nanotechnology during last the 20 years for the treatment of neurodegenerative diseases.

Effect of kaempferol on the transgenic Drosophila model of Parkinson's disease.

The present study was aimed to study the effect of kaempferol, on the transgenic Drosophila model of Parkinson's disease. Kaempferol was added in the diet at final concentration of 10, 20, 30 and 40 µM and the effect was studied on various cognitive and oxidative stress markers. The results of the study showed that kaempferol, delayed the loss of climbing ability as well as the activity of PD flies in a dose dependent manner compared to unexposed PD flies. A dose-dependent reduction in oxidative stress markers was also observed. Histopathological examination of fly brains using anti-tyrosine hydroxylase immunostaining has revealed a significant dose-dependent increase in the expression of tyrosine hydroxylase in PD flies exposed to kaempferol. Molecular docking results revealed that kaempferol binds to human alpha synuclein at specific sites that might results in the inhibition of alpha synuclein aggregation and prevents the formation of Lewy bodies.

Ropinirole silver nanocomposite attenuates neurodegeneration in the transgenic Drosophila melanogaster model of Parkinson's disease.

Parkinson's disease (PD) is a progressive neurodegenerative disease due to the degeneration of dopaminergic neurons in substantia nigra pars compacta of the mid brain. The present study investigates the neuro-protective role of synthesized ropinirole silver nanocomposite (RPAgNC) in Drosophila model of PD. α-synuclein accumulation in the brain of flies (PD flies) leads to the damage of dopaminergic neurons, dopamine depletion, impaired muscular coordination, memory decline and increase in oxidative stress. Ingestion of the RPAgNC by Drosophila significantly prevented the neuronal degeneration compared to only ropinirole. The results confirm that the RPAgNC exerts more neuro-protective effect compared to dopamine agonist i.e. ropinirole as such drug in experimental PD flies. This article is part of the special issue entitled 'The Quest for Disease-Modifying Therapies for Neurodegenerative Disorders'.

Effect of Majun Baladur on life span, climbing ability, oxidative stress and dopaminergic neurons in the transgenic Drosophila model of Parkinson's disease.

The effect of a poly herbal drug Majun Baladur (MB) was studied on the transgenic Drosophila melanogaster expressing human alpha synuclein in the neurons (PD flies). The equivalents of recommended dose for human were established for 20 g of fly food i.e. 0.0014, 0.0028, 0.0042 and 0.0056 g per 20 g of diet. The PD flies were allowed to feed on it for 24 days before performing the assays. The exposure to MB increased the life span and improves the activity of PD flies. The PD flies exposed to 0.0014, 0.0028, 0.042 and 0.0056 g of MB showed a dose dependent significant delay of 1.47, 1.88, 2.52 and 3.05 folds in the climbing ability compared to unexposed PD flies. A dose dependent significant decrease of 1.38, 1.45, 1.48 and 1.65 folds in TBARS; 1.08, 1.11, 1.17 and 1.20 folds in the GST activity; 1.20, 1.28, 1.39 and 1.52 folds in the PC content; 1.43, 1.53, 1.65 and 1.79 folds in the Caspase-9 activity; 1.21, 1.31, 1.53 and 1.64 folds in the activity of Caspase-3 and 1.24, 1.42, 1.50 and 1.79 folds in the activity of catalase; 1.50, 1.63, 1.88 and 2.06 folds in the activity of SOD in PD flies exposed to 0.0014, 0.0028, 0.042 and 0.0056 g of MB, respectively. A significant dose dependent increase of 1.20, 1.29, 1.33 and 1.44 folds in as NPSH content was observed in PD flies exposed to 0.0014, 0.0028, 0.042 and 0.0056 g of MB, respectively. The exposure to MB protects the loss of dopaminergic neurons as is evident by immunohistochemistry. It is concluded that MB is potent in reducing the PD symptoms being mimicked in the transgenic flies.

Effect of Genistein on the Transgenic Drosophila Model of Parkinson's Disease.

Parkinson's disease (PD) is the second-most common neurodegenerative disorder and is characterized by the degeneration of dopaminergic neurons in the substantia nigra pars compacta. Oxidative stress has also been linked with the progression of PD, hence the involvement of a natural plant product could offer neuroprotection. The present study deals with the effect of genistein on the transgenic flies expressing normal human alpha synuclein panneurally. The PD flies were exposed to 10, 20, 30, and 40 µM of genistein (mixed in diet) for 24 days. A significant dose-dependent increase in the life span and delay in the loss of climbing ability were observed in the PD flies exposed to genistein (p < .05). A significant dose-dependent decrease in oxidative stress markers and increase in dopamine content were observed in PD flies exposed to genistein. However, the exposure of genistein did not inhibit the expression of α-synuclein in the brains of PD flies.

Therapeutic potential of luteolin in transgenic Drosophila model of Alzheimer's disease.

A transgenic fly line expressing wild type human Aß42 were exposed to luteolin mixed in diet at final concentration of 5, 10, 15 and 20µM. The climbing assay, activity pattern, life span, aversive phototaxis suppression assay (APS) along with the estimation of protein carbonyl content (PCC), glutathione-S-transferase (GSTs) activity, glutathione (GSH) content, lipid peroxidation (LPO), acetylcholinesterase activity (AChE), superoxide dismutase (SOD) activity, catalase (CAT) activity, caspase 3 and 9 activities in the brain of treated as well as untreated AD flies (Positive control) were studied. Histopathology of Drosophila brain sections was done by performing thioflavin-S, Bielschowsky's silver staining and toluidine blue staining. A dose-dependent increase in the life span, delay in the loss of climbing ability as well as activity was observed in AD flies exposed to luteolin compared to unexposed AD flies. A dose-dependent reduction in LPO, PCC, GST, AChE, SOD, CAT, caspase 9 and caspase 3 activity and an increase in the GSH content was also observed. Histopathological examination of fly brains using thioflavin-S and silver staining has revealed a significant dose-dependent reduction in the expression of Aß42 peptides in AD fly groups exposed to 10, 15 and 20µM of luteolin. No gross morphological changes were observed in the brain sections of AD and control flies stained with toluidine blue. Molecular docking results have revealed that luteolin binds to AChE and Aß42 at specific sites that might result in the inhibition of AChE and disaggregation/prevention of Aß42 plaque formation.

Protective Effect of Kaempferol on the Transgenic Drosophila Model of Alzheimer's Disease.

BACKGROUND: Alzheimer's disease (AD) is characterized by the accumulation and deposition of ß-amyloid peptides leading to a progressive neuronal damage and cell loss. Besides several hypotheses for explaining the neurodegenerative mechanisms, oxidative stress has been considered to be one of them. Till date, there is no cure for AD, but the pathogenesis of the disease could be delayed by the use of natural antioxidants. In this context, we decided to study the effect of kaempferol against the transgenic Drosophila expressing human amyloid beta-42. METHOD: The AD flies were allowed to feed on the diet having 10, 20, 30 and 40µM of kaempferol for 30 days. After 30 days of exposure, the amyloid beta flies were studied for their climbing ability and Aversive Phototaxis Suppression assay. Amyloid beta flies head homogenate was prepared for estimating the oxidative stress markers, Caspase and acetylcholinesterase activity. RESULTS: The results of the present study reveal that the exposure of AD flies to kaempferol delayed the loss of climbing ability, memory, reduced the oxidative stress and acetylcholinesterase activity. CONCLUSION: Kaempferol could be used as a possible therapeutic agent against the progression of the Alzheimer's disease.

Protective effect of tangeritin in transgenic Drosophila model of Parkinson's disease.

Parkinson's disease (PD) is a neurodegenerative disorder caused due to the loss of dopaminergic neurons in substantia nigra region of midbrain. The disease is characterized by the accumulation of alpha-synuclein into depositions known as lewy bodies. Till date there is no cure for PD but the limited number of medications may provide temporary relief from the PD symptoms. Flavonoids are a group of polyphenols found in plants. The health benefits of flavonoids have been universally accepted. Tangeritin is a pentamethoxy flavone found in the peels of Mandarin oranges (Citrus reticulata). The present study was conducted to study the effect of tangeritin on the symptoms of PD exhibited by the PD model transgenic flies (Drosophila melanogaster). Tangeritin at a final concentration of 5, 10 and 20 microM was added to the diet and the flies were allowed to feed on it for 24 days. At the same time other set of PD flies were allowed to feed on a diet having 10-3 M of L-Dopa. The effect of tangeritin was studied on the activity pattern, climbing ability, dopamine content, oxidative stress markers (lipid peroxidation, reduced glutathione, glutathione-S-transferase, protein carbonyl content and monoamine oxidase activity) and on the histopathology of the brain of PD model flies. The study showed that the exposure of PD flies to different doses of tangeritin showed a marked delay in the loss of climbing ability and increase in the dopamine content. Tangeritin also showed a reduction in various oxidative stress markers. Hence it is concluded that tangeritin showed a marked reduction in the PD symptoms and thus could be of great importance for further research in treating PD.

Protective effect of capsaicin against methyl methanesulphonate induced toxicity in the third instar larvae of transgenic Drosophila melanogaster (hsp70-lacZ)Bg9.

Capsaicin (trans-8-methyl-N-vanillyl-6-nonenamide) is the main component in hot peppers, including red chili peppers, jalapenos, and habanero, belonging to the genus Capsicum. Capsaicin is a potent antioxidant that interferes with free radical activities. In the present study, the possible protective effect of capsaicin was studied against methyl methanesulphonate (MMS) induced toxicity in third instar larvae of transgenic Drosophila melanogaster (hsp70-lacZ)Bg9. The third instar was allowed to feed on the diet having different doses of capsaicin and MMS separately and in combination. The results suggested that the exposure of third instar larvae to the diet having MMS alone showed significant hsp70 expression as well as tissue DNA and oxidative damage, whereas the larvae feed on the diet having MMS and capsaicin showed a decrease in the toxic effects for 48-h of exposure. In conclusion, capsaicin showed a dose-dependent decrease in the toxic effects induced by MMS in the third instar larvae of transgenic Drosophila melanogaster.

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 Parkinson's disease.