RESUMO

Agents that suppress the toxic effect of arecoline (a chemical present in the Areca nut fruit) have become a need of the hour owing to its several harmful effects on human beings. Although some drug molecules have been developed for this purpose, yet, simple, easy to prepare, and economical molecules with remarkable potency are still a challenge to design. The present work thus becomes important as it involves the synthesis of a new charge transfer complex (CTC) material, which has, for the first time, been screened to investigate its effect on the toxic effects of arecoline. The newly designed material (CL), which is generated from the reaction between 2,4,6-trinitrophenol (TNP) and pyrazole (PYZ), has been crystallized by a slow evaporation method and characterized by employing spectral studies including single crystal X-ray crystallography. Spectrophotometry studies with the inclusion of the Benesi-Hildebrand equation reveal 1 : 1 stoichiometry and physical parameters of CL. Assays were used for determining the protective effect of CL against arecoline. CL was found to (dose-dependently) decrease ß-galactosidase activity, damage in tissue and DNA damage caused by arecoline (80 µM) in the third-instar larvae of the transgenic Drosophila melanogaster (hsp70-lacZ)Bg9. The possible mechanism of this effect was explored through fluorescence and UV-vis spectroscopy. The possibility of suppression of arecoline action on the muscarinic acetylcholine receptor 1-G11 protein complex (found in the cell membrane) in the presence of CL was studied theoretically by molecular docking. Density functional theory (DFT) also theoretically supported various aspects of the designed material concerning the energy profile of the orbitals (HOMO-LUMO) as well as the energy minimized structure. Furthermore, time dependent (TD) DFT corroborated the electronic properties of the designed material.

Assuntos

Arecolina , Drosophila melanogaster , Animais , Animais Geneticamente Modificados , Arecolina/toxicidade , Drosophila melanogaster/metabolismo , Proteínas de Choque Térmico HSP70/metabolismo , Óperon Lac , Larva , Simulação de Acoplamento Molecular , Tomografia Computadorizada por Raios X

RESUMO

The novel Mannich base benzimidazole derivative (CB-1), 1-((1H-benzo[d]imidazol-1-yl)(3-chlorophenyl)methyl)-3-phenylurea) has been designed and synthesized by reacting benzimidazole, 3-chloro benzaldehyde, and N-Phenyl urea. CB-1 has been characterized by UV- Visible, FTIR, and 1H NMR. CB-1 was explored to study the interaction with the most abundant blood protein which involved in the role of transport of molecules (drugs), human serum albumin (HSA). Fluorescence results are evident for the presence of both dynamic and static quenching mechanisms in the binding of CB-1 to HSA. Antimicrobial screening were carried out against three bacteria and three fungi pathogens employing disc diffusion method. Molecular docking using AutoDock Vina tool further confirms the experimental binding interactions obtained from fluorescence. Density functional theory (DFT) with B3LYP/6-311G++ basis set was used for correlating theoretical data and obtaining optimized structures of CB-1 along with reactants with molecular electrostatic potential (MEP) map and HOMO→LUMO energy gap calculation. HIGHLIGHTSThe novel Mannich base benzimidazole derivative (CB-1) has been designed and synthesized by Mannich reaction.CB-1 has been characterized by UV- Visible, FTIR, and 1H NMR.Fluorescence quenching reveals that HSA binds to CB-1 via aromatic residues, which is corroborated by molecular docking.Antifungal and antibacterial activity was evaluated in comparison to Nystatin and Gentamicin standard drugs, respectively.DFT calculations support experimental data and provide HOMO-LUMO energy gap.Communicated by Ramaswamy H. Sarma.

RESUMO

In the present study geraniol at the final concentration of 10, 20, 30, and 40 µM was mixed in the diet along with 80 µM of arecoline and the third instar larvae of transgenic Drosophila melanogaster (hsp70-lacZ) Bg9 were allowed to feed on it for 24 hrs. After the exposure of 24 hrs the larvae were subjected to ONPG, X-gal, trypan blue exclusion test, oxidative stress markers and apoptotic and comet assays. The exposure of larvae to geraniol showed a dose dependent decrease in the activity of ß-galactosidase, tissue damage and oxidative stress markers. A dose dependent decrease in apoptosis and DNA damage was also observed. Molecular docking studies also support the protective role of geraniol against the arecoline induced toxicity. The results suggest that geraniol is potent in reducing the toxicity induced by arecoline in the third instar larvae of transgenic Drosophila melanogaster (hsp70-lacZ) Bg9.

Assuntos

Apoptose/efeitos dos fármacos , Arecolina/toxicidade , Dano ao DNA/efeitos dos fármacos , Drosophila melanogaster/efeitos dos fármacos , Sequestradores de Radicais Livres/farmacologia , Proteínas de Choque Térmico HSP70/genética , Estresse Oxidativo/efeitos dos fármacos , Terpenos/farmacologia , Monoterpenos Acíclicos , Animais , Animais Geneticamente Modificados , Antioxidantes/metabolismo , DNA/metabolismo , Relação Dose-Resposta a Droga , Drosophila melanogaster/enzimologia , Drosophila melanogaster/genética , Óperon Lac , Larva/efeitos dos fármacos , Larva/enzimologia , Larva/genética , Simulação de Acoplamento Molecular

RESUMO

Arecoline is the key component of areca nut and has been suggested as a carcinogenic agent. In the present study, the third instar larvae of transgenic Drosophila melanogaster (hsp70-lacZ) Bg9 were allowed to feed on a diet having 5, 10, 20, 40 and 80 µM arecoline for 24 h. After the completion of 24 h, the larvae were subjected to ONPG assay, X-gal staining, trypan blue exclusion test, oxidative stress markers, and apoptotic and comet assays. A dose-dependent increase in the ß-galactosidase activity, tissue damage, glutathione-S-transferase (GST) activity, lipid peroxidation assay, monoamine oxidase (MAO), caspase-9 and 3, protein carbonyl content (PCC), apoptotic index, and DNA damage and decrease in glutathione (GSH) content, delta aminolevulinic acid dehydrogenase (δ-ALA-D), and acetylcholinesterase (AChE) activity were observed in the larvae exposed to 20, 40 and 80 µM arecoline. The results suggest that arecoline is toxic at 20, 40, and 80 µM toward the third instar larvae of transgenic Drosophila melanogaster (hsp70-lacZ) Bg9 . Arecoline did not show any toxic effects at 5 and 10 µM.

RESUMO

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.

Assuntos

Anticarcinógenos/farmacologia , Capsaicina/farmacologia , Drosophila melanogaster/efeitos dos fármacos , Metanossulfonato de Metila/antagonistas & inibidores , Acetilcolinesterase/metabolismo , Animais , Animais Geneticamente Modificados , Dano ao DNA/efeitos dos fármacos , Larva/efeitos dos fármacos

RESUMO

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.

Assuntos

Antioxidantes/administração & dosagem , Ácido Ascórbico/administração & dosagem , Encéfalo , Proteínas de Drosophila/metabolismo , Transtornos Neurológicos da Marcha/prevenção & controle , Doença de Parkinson/patologia , Fatores Etários , Animais , Animais Geneticamente Modificados , Antiparkinsonianos/uso terapêutico , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Encéfalo/patologia , Modelos Animais de Doenças , Progressão da Doença , Relação Dose-Resposta a Droga , Drosophila , Transtornos Neurológicos da Marcha/etiologia , Humanos , Levodopa/uso terapêutico , Proteínas de Membrana/genética , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Doença de Parkinson/complicações , Doença de Parkinson/tratamento farmacológico , Doença de Parkinson/genética , Fatores de Transcrição/metabolismo , alfa-Sinucleína/genética

RESUMO

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.