Putative Involvement of Thiol Protease Inhibitor in the Function of Alzheimer Drug

The intermolecular structure gets altered when drug-protein interaction takes place. It brings about alterations in the conformation of protein. An acetyl cholinesterase inhibitor (AChE) is the most used drug for patients who are suffering from Alzheimer’s disease to curb its instigated symptoms. So, it is used as first-line defense in the insightful symptoms. This study is of concern with the interaction of cystatin purified from buffalo brain with its simple tri-step procedure including alkaline action, ammonium sulfate fractionation, and gel filtration chromatography on Sephadex G-75 with % yield of 64.13 and fold purification of 384.7. The inhibitor (brain cystatin (BC)) showed a single papain inhibitory peak drifted as single band on native PAGE; this purified inhibitor was interacted with donepezil to analyze the side effect of this drug since cystatin is an important regulatory protein that maintains the protease antiprotease balance. The conformational change was predicted when the UV spectra of cystatin was analyzed in the presence of donepezil contextual with the fluorescence spectra, but the fluorescence spectra showed 40 nm of red shift suggesting the change on interaction leading to a conclusion that donepezil is pertinent to imbalance to protease and antiprotease inhibitor perhaps the side effect of drug

Assessment of fish oil to check the stability and meat quality of some commercially available tin packed fish in Islamabad, Pakistan

Fish contains all the essential nutrients which is important for human health so it is important to analyse all the nutrients present in tin packed fish meat. Keeping in view its importance present study is conducted on the assessment of fish oil to check the stability and quality of fish meat by proximate analysis of commercially available tin packed fish. Four tin packed fish species i.e., Sardine (Sardinella longiceps), Red salmon (Oncorhynchus nerka), Pink salmon (Oncorhynchus gorbuscha) and Skipjack tuna (Katsuwonus pelamis) were collected from different supermarkets of Islamabad, Pakistan. Proximate analysis viz., crude protein, crude fat, moisture and ash contents of the fish meat has been done to evaluate the meat quality. The antioxidant activity in oil was also analysed by FRAP assay. The result indicated that maximum percentage of moisture i.e., 78.61% present in Skipjack Tuna meat, Pink Salmon contain highest percentage of crude protein i.e., 70.00%, Red Salmon contain highest percentage of crude fat i.e., 30.00% while Sardine and Skipjack Tuna contains highest percentage of ash contents i.e., 8.00% and the total antioxidant capacity (uM) is higher in oil of Red Salmon (24.35%) followed by Sardines (14.78%), Skipjack Tuna (9.86%) and Pink Salmon (9.48%). It was concluded that the fish meat after thermal processing contains suitable percentage of crude protein, crude fats, and moisture and ash contents

Studies on Interaction of Buffalo Brain Cystatin with Donepezil: An Alzheimer's Drug

When drugs bind to a protein, the intramolecular structures can be altered, resulting in conformational change of the protein. Donepezil, an Acetyl Cholinesterase inhibitor (AChE), is commonly prescribed to patients with Alzheimer's disease (AD) to enhance cholinergic neurotransmission. It is the “first-line” agents in the treatment of Alzheimer's disease used to improve cognitive function in the disease. In the present study, a cysteine protease inhibitor (cystatin) has been isolated from buffalo brain using alkaline treatment, 40 to 60% ammonium sulphate fractionation and gel filtration chromatography on Sephadex G-75 with % yield of 64.13 and fold purification of 384.7. The purified inhibitor (Buffalo Brain Cystatin, (BBC)) was eluted as a single papain inhibitory peak which migrated as single band on native PAGE; however, on SDS-PAGE with and without beta mercaptoethanol (βME) BBC gave two bands of M W 31.6 and 12.4 KDa, respectively. The molecular weight determined by gel filtration came out to be 43.6 KDa. The UV spectra of cystatin on interaction with donepezil suggested a conformational change in the protein. The fluorescence spectra of BC-donepezil composite show structural changes indicating 40 nm red shift with significant increase in fluorescence intensity of cystatin in the presence of donepezil representing an unfolding of cystatin on interaction, which is an indication of side effect of donepezil during the use of this drug

Effect of dietary n-3 and n-6 fatty acids on tobramycin induced nephrotoxicity

691-697Post fish oil(n-3 fatty acids) treatment (5mg/kg/day for 12 days) was effective in bringing the reversal of tobramycin (160mg/kg/day,ip for 12 days) induced nephrotoxicity in albino rats as was evident by normal urea, creatinine, cholesterol and inorganic phosphate levels in the serum of the treatment group compared with group receiving tobramycin only. The return of normal levels of alkaline and acid phosphatase in kidney homogenates of post fish oil treatment group also indicated the beneficial effect of dietary n-3 fatty acids(fish oil) more than n-6 fatty acids(olive oil).The results suggest that oral supplements of dietary n-3 fatty acids (fish oil) for nearly two weeks after tobramycin exposure is more beneficial than n-6 fatty acids (olive oil) as it results in reversal of nephrotoxicity induced by tobramycin

Methotrexate binding causes structural and functional changes in lung cystatin

Regulation of cysteine proteinases and their inhibitors is of utmost importance in diseases like lung cancer, chronic inflammatory conditions such as asthma, emphysema, and idiopathic pulmonary fibrosis. Protease-antiprotease imbalance accelerates disease progression. In the present study, the effect of antineoplastic and antirheumatic drug methotrexate (MTX) on lung cystatin (a cysteine protease inhibitor) was studied to explore drug induced changes in functional and structural integrity of the protein. The basic binding interaction was studied by UV-absorption, FT-IR and fluorescence spectroscopy. The quenching of protein fluorescence confirmed the binding of MTX with goat lung cystatin (GLC-I). Stern-Volmer analysis of MTX-GLC-I system at different temperatures indicates the presence of static component in the quenching mechanism. The thermodynamic parameters ΔH0 and ΔS0 were -3.8 kJ/mol and 94.97 J•mol-1•K-1, respectively, indicating that both hydrogen bonds and hydrophobic interactions played a major role in the binding of MTX to GLC-I. Methotrexate (7 µM) caused complete inactivation of lung cystatin after 6 hours. The results of FT-IR spectroscopy reflect perturbation of the goat lung cystatin on interaction with MTX. Methotrexate induced loss of function change in the inhibitor could provide a rationale for the off target tissue injury caused by the drug and for the design of agents against such an injury

Antidiabetic and antioxidant activity of Annona squamosa extract in streptozotocin-induced diabetic rats

Introduction: The aim of the study is to analyse the antioxidant effect of oral administration of aqueous extract of Annona squamosa (A. squamosa) leaf on blood glucose, haemoglobin, glycosylated haemoglobin, plasma insulin, antioxidant enzymes and lipid peroxidation in liver and kidney to streptozotocin (STZ)-induced diabetic rats. Methods: Aqueous extract of A. squamosa on blood glucose, haemoglobin, glycosylated haemoglobin, plasma insulin, serum lipid and the levels of lipid peroxides and antioxidant enzymes, such as catalase, superoxide dismutase, glutathione peroxidase and reduced glutathione, were examined in the liver and kidney tissues of control and experimental groups. Results: Oral administration of A. squamosa aqueous extract to diabetic rats for 30 days significantly reduced the levels of blood glucose, lipids and lipid peroxidation, but increased the activities of plasma insulin and antioxidant enzymes, like catalase, superoxide dismutase, reduced glutathione and glutathione peroxidase. Conclusion: The A. squamosa aqueous extract supplementation is useful in controlling the blood glucose level, improves the plasma insulin, lipid metabolism and is beneficial in preventing diabetic complications from lipid peroxidation and antioxidant systems in experimental diabetic rats; therefore, it could be useful for prevention or early treatment of diabetes mellitus

Biochemical effects of <i style="">Nigella sativa</i> L seeds in diabetic rats

745-748Oral administration of ethanol extract of N. sativa seeds (300 mg/kg body weight/day) to streptozotocin induced diabetic rats for 30 days significantly reduced the elevated levels of blood glucose, lipids, plasma insulin and improved altered levels of lipid peroxidation products (TBARS and hydroperoxides) and antioxidant enzymes like catalase, superoxide dismutase, reduced glutathione and glutathione peroxidase in liver and kidney. The results confirm the antidiabetic activity of N. sativa seeds extract and suggest that because of its antioxidant effects its administration may be useful in controlling the diabetic complications in experimental diabetic rats

Physico-chemical and In silico analysis of a phytocystatin purified from Brassica juncea cultivar RoAgro 5444

This study describes the isolation and purification of a phytocystatin from seeds of Brassica juncea (B.juncea), (cultivar RoAgro 5444), an important oilseed crop with pervasive agricultural and economic ramifications. The protein was purified by gel filtration chromatography with 24.3% yield and 204 fold purification and visualised by 2D gel electrophoresis. The 18.1kDa mustard cystatin was highly specific for cysteine proteinases. The plant cystatin inhibited cathepsin B, confirming its role in conferring pest resistance. The inhibitor was highly stable over a pH range of 3-10 and retained significant inhibitory potential till 70ºC. The stoichiometry of its interaction with papain, determined by Isothermal Calorimetry suggests a 1:1 complex. Secondary structural elements calculated by far UV circular dichroism (CD) spectroscopy show 18.8% α helical and 21% β sheet structure. The protein was a non-competitive inhibitor of thiol proteinases. The stoke’s radius and frictional co-efficient were used to describe the shape and size of the protein.Homology modelling and docking studies proposed a prototype illustrating the Brassica phytocystatin mediated papain inhibition. Molecular dynamics (MD) study revealed prominent stability of papain-phytocystatin complex during 100 nanoseconds (ns) long simulation. Detailed results identify the mustard cystatin as an important member of the phytocystatin family.The accepted manuscript in pdf format is listed with the files at the bottom of this page. The presentation of the authors' names and (or) special characters in the title of the manuscript may differ slightly between what is listed on this page and what is listed in the pdf file of the accepted manuscript; that in the pdf file of the accepted manuscript is what was submitted by the author

Beneficial effects of Annona squamosa extract in streptozotocin-induced diabetic rats

Introduction: The present study investigated the possible therapeutic effects of Annona squamosa (A. squamosa) extract on certain biochemical markers in streptozotocin (STZ) -induced diabetes mellitus in rats. Methods: The effects of an aqueous extract of A. squamosa leaves on blood glucose, insulin, C-peptide, albumin, albumin/globulin ratio, urea, uric acid and creatinine and the activities of diagnostic marker enzymes aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase and gamma-glutamyl transpeptidase were examined in the plasma, liver and kidney tissues of control and experimental groups. Results: Oral administration of A. squamosa (300 mg/kg) aqueous extract to diabetic rats for 30 days significantly reduced blood glucose, urea, uric acid and creatinine, but increased the activities of insulin, C-peptide, albumin, albumin/globulin ratio and restored all marker enzymes to near control levels. Conclusion: The present results shown that A. squamosa extract has an antihyperglycaemic effect and consequently may alleviate liver and renal damage associated with STZ-induced diabetes mellitus in rats