Vascular Endothelial Growth Factor Receptor, fms-Like Tyrosine Kinase-1 (Flt-1), as a Novel Binding Partner for SARS-CoV-2 Spike Receptor-Binding Domain.

Angiotensin-converting enzyme 2 (ACE2) and neuropilin 1, a vascular endothelial growth factor (VEGF) receptor, were identified to bind to the SARS-CoV-2 spike receptor-binding domain (spike RBD). In silico analysis based on 3D structure, multiple sequence alignment, and molecular docking of second domain of soluble Flt-1 (sFlt-1) and spike RBD revealed structural similarities, sequence homology, and protein-protein interaction. Interaction and binding of recombinant spike RBD (rspike RBD) and recombinant sFlt-1 (rsFlt-1) in vitro induced a conformational change, as revealed by spectrofluorimetric data, with increased fluorescence intensity in emission spectra as compared to either of the proteins alone. Results on ELISA confirmed the binding and cross-reactivity of rspike-RBD and rsFlt-1 as determined by using either specific antibodies towards each protein or immunized human serum. We found that polyclonal or monoclonal anti-spike RBD antibodies can recognize either rsFlt-1 or rspike RBD, showing cross-reactivity for the two proteins in a dose-dependent binding response. Recognition of bound rspike RBD or rsFlt-1 by anti-Flt-1 or anti-spike RBD antibodies, respectively, as observed by immunoblotting, further confirmed interaction between the two proteins. Immunoprecipitation and immunoblot analysis demonstrated the identification of rspike RBD binding to the Flt-1 receptor on A549 cells. Further, the binding of rspike RBD to Flt-1 receptor was shown using immunofluorescence on 2D-culture or 3D-spheroid of MDA-MB-231 cells, which over-express Flt-1 receptor. Together, our study concludes that the Flt-1 receptor is a novel binding partner for SARS-CoV-2 spike RBD.

Comparative assessment of cognitive impairment and oxidative stress markers among vitamin D insufficient elderly patients with and without type 2 diabetes mellitus (T2DM).

BACKGROUND: Disorders of mental health are known to affect cognitive functions, hence called as cognitive disorders. Impaired glucose metabolism, insulin resistance, vitamin-D deficiency and oxidative stress are some of the key early events reported to be involved in the pathogenesis of most common cognitive disorders, which include Alzheimer's disease. Type-2 diabetes mellitus (T2DM) is one of the known contributing factors of cognitive impairment and dementia. METHODS: A cross sectional study was carried out in 145 subjects, who were assessed for cognitive function by modified mini mental status examination (3MS). In addition, measurement of fasting blood sugar (FBS), fasting insulin, HbA1c, lipid profile, vitamin D and oxidative markers was performed. Participants were divided into different groups based on (a) vitamin D insufficiency and sufficiency; (b) diabetic and non-diabetic with and without cognitive impairment. RESULTS: The study included a total of 145 subjects; 51 males and 94 females and the mean age was 68.24±9.70 years. Among diabetics with vitamin D insufficiency, 35 subjects (71.43%) had cognitive impairment, but, among non-diabetics with vitamin D insufficiency, 27 subjects (62.79%) had cognitive impairment. Chi square test showed no significant association between diabetes, vitamin D insufficiency and cognitive impairment. Glutathione peroxidase (GPx) and superoxide dismutase (SOD) levels were non-significantly lower in cognition-impaired subjects, when compared to cognition normal subjects in diabetics with vitamin D insufficiency. CONCLUSION: Our study showed that cognitive impairment is more predominant in individuals with diabetes. However, our study did not find any significant relationship between T2DM, vitamin D deficiency, cognitive impairment, and oxidative stress. A significant association was found only with GPx and 3MSE score in vitamin D insufficient non-diabetics.

Diet-induced hypercholesterolemia alters liver glycosaminoglycans and associated-lipoprotein receptors in rats.

Glycosaminoglycans (GAGs) play an important role in lipoprotein metabolism. In liver, it facilitates the uptake of remnants through receptor-independent endocytosis. However, changes in liver GAGs during diet-induced hypercholesterolemia with normal levels of fat feeding are unknown. Present paper highlights the effect of diet-induced hypercholesterolemia with normal levels (5%) of fat on liver GAGs and other associated lipoprotein receptors. Hypercholesterolemia was induced in rats by feeding diet supplemented with 0.5% cholesterol and 0.125% bile salts. Hypercholesterolemia showed significantly decreased GAGs of both heparan sulfate (HS) and chondroitin sulfate/dermatan sulfate (CS/DS) classes of molecules. Quantitative real-time polymerase chain reaction analysis of GAG biosynthetic enzymes and other genes revealed significant changes in expression profile. The decrease in GAGs was prevented by simvastatin treatment; a drug that inhibits endogenous cholesterol synthesis that was used as a positive control in our study. Furthermore, there was a comparatively decreased binding of GAGs from hypercholesterolemic rats to lipoprotein lipase. LRP1 which plays a major role in lipoprotein uptake was also significantly decreased, and it was attenuated in simvastatin-treated hypercholesterolemic rats. Furthermore, LDLR and ApoE were also decreased significantly in liver of hypercholesterolemic rats. Thus, diet-induced hypercholesterolemia results in dysregulation of cholesterol homeostasis apparently through changes in GAGs in conjunction with other associated players.

Anti-diabetic effect of dietary mango (Mangifera indica L.) peel in streptozotocin-induced diabetic rats.

BACKGROUND: In the present study, the composition of mango peel powder (MPP) collected from the mango pulp industry was determined and the effect of MPP on ameliorating diabetes and its associated complications was studied. RESULTS: Mango peel was rich in polyphenols, carotenoids and dietary fibre. Peel extract contained various bioactive compounds and was found to be rich in soluble dietary fibre. Peel extract exhibited antioxidant properties and protected against DNA damage. Therefore, the effect of peel on ameliorating diabetes was investigated in a rat model of diabetes. A significant increase in urine sugar, urine volume, fasting blood glucose, total cholesterol, triglycerides and low density lipoprotein, and decrease in high density lipoprotein were observed in the rats; however, these parameters were ameliorated in diabetic rats fed with diet supplemented with mango peel at 5% and 10% levels in basal diet. Treatment of diabetic rats with MPP increased antioxidant enzyme activities and decreased lipid peroxidation in plasma, kidney and liver compared to untreated diabetic rats. Glomerular filtration rate and microalbuminuria levels were ameliorated in MPP treated diabetic group. CONCLUSIONS: Mango peel, a by-product, can be used as an ingredient in functional and therapeutic foods.

Effect of curcumin and quercetin on lysosomal enzyme activities in streptozotocin-induced diabetic rats.

BACKGROUND & AIMS: Diabetes causes impairment of various enzyme activities in the physiological system, including lysosomal enzymes. The effect of feeding curcumin, quercetin and aminoguanidine on lysosomal enzyme activities viz., N-acetyl-ß-d-glucosaminidase, ß-d-glucuronidase, ß-d-galactosidase and acid phosphatase were studied in different tissues of streptozotocin-induced diabetic rats. METHOD: Rats were divided into four control groups and four diabetic groups. Experimental groups were fed with diet supplemented with curcumin (0.5%) or quercetin (0.1%) or aminoguanidine (0.05%). Lysosomal enzyme activities were determined in various tissues. RESULTS: The specific activity of N-acetyl-ß-d-glucosaminidase in liver of diabetic rats was decreased when compared to control rats and was ameliorated with curcumin and quercetin treatment by 67% and 78%, respectively. On the other hand, ß-d-glucuronidase activity was higher in the brain of diabetic rats (0.90 ± 0.04 nmol/mg protein/min), when compared to control rats (0.45 ± 0.02 nmol/mg protein/min) and was decreased in curcumin (0.75 ± 0.05 nmol/mg protein/min) and quercetin (0.74 ± 0.11 nmol/mg protein/min) treated rats. ß-d-galactosidase activity in spleen of curcumin and quercetin fed diabetic group rats was ameliorated by 68% and 58%, respectively, in comparison to diabetic rats. Acid phosphatase activity in diabetic rats decreased in testis when compared to control. CONCLUSION: Curcumin and quercetin feeding modulated lysosomal enzyme activities in different tissues during diabetes and the effect was comparable to well-known anti-glycative agent - aminoguanidine.

Banana (Musa sp. var. elakki bale) flower and pseudostem: dietary fiber and associated antioxidant capacity.

Banana flower (BF) and pseudostem (PS) are byproducts of banana cultivation and are known to have health beneficial effects. The main objective of this study was to evaluate the dietary fiber composition and antioxidant effect of BF and PS. In the present study, BF and PS were found to be rich in dietary fiber (65.6 ± 1.32 and 28.8 ± 0.98%, respectively). Dietary fiber fractions were extracted and characterized in terms of sugar profile, and antioxidant activities were determined. BF and PS fractions were rich in sugars and showed wide diversity with respect to the nature of the sugars. Hemicellulose A fraction of BF showed high amounts of total polyphenols and total antioxidants, which were 121.8 ± 1.9 and 39.03 ± 0.118 µg/mg extract, respectively. HPLC analysis showed the presence of phenolic acids in hemicellulose A and B fractions of BF. These results indicate that BF and PS are rich sources of dietary fiber associated with polyphenols, which could promote health beneficial effects.

Tinospora cordifolia consumption ameliorates changes in kidney chondroitin sulphate/dermatan sulphate in diabetic rats.

Diabetes is known to alter kidney extracellular matrix (ECM) components. Chondroitin sulphate (CS)/dermatan sulphate (DS), an ECM component, which plays an essential role in kidney is altered during diabetes. The focus of this study has been to examine the effect of Tinospora cordifolia (TC) consumption, a potent plant widely used to treat diabetes, on kidney CS/DS. Experimentally induced diabetic rats were fed with diet containing TC at 2·5 and 5 % levels and the effect of it on kidney CS/DS was examined. The CS/DS content and CS:heparan sulphate ratio which was decreased during diabetic condition were ameliorated in TC-fed groups. Disaccharide composition analysis of CS/DS by HPLC showed that decreases in 'E' units and degree of sulphation were modulated in 5 % TC-fed groups. Apparent molecular weight of purified CS/DS from the control rat kidney was found to be 38 kDa which was decreased to 29 kDa in diabetic rat kidney. Rats in 5 % TC-fed groups showed chain length of 38 kDa akin to control rats. Expression of chondroitin 4-O-sulfotransferase-1, dermatan 4-O-sulfotransferase-1 and N-acetylgalactosamine 4 sulphate 6-O-sulfotransferase, enzymes involved in the synthesis of 'E' units which was reduced during diabetic condition, was significantly contained in the 5 % TC-fed group. Purified CS/DS from 5 % TC-fed group was able to bind higher amounts of ECM components, namely type IV collagen and laminin, when compared with untreated diabetic rats. The present results demonstrate that consumption of a diet containing TC at the 5 % level modulates changes in kidney CS/DS which were due to diabetes.

Enhancement of soluble dietary fibre, polyphenols and antioxidant properties of chapatis prepared from whole wheat flour dough treated with amylases and xylanase.

BACKGROUND: Chapati preparation involves various processing steps such as mixing the flour into dough, sheeting and baking. During these processing steps, flour components are likely to undergo changes in their nutrient and polyphenol composition and their antioxidant properties due to phenol-mediated crosslinking of proteins and carbohydrates. Therefore, in the present study, changes in nutritional, nutraceutical and antioxidant properties of chapatis prepared from doughs treated with amylases and xylanase were determined. RESULTS: An increase in insoluble dietary fibre content and a decrease in soluble polyphenol content were observed during preparation of control chapatis from whole wheat flours. However, significant increases in soluble dietary fibre and soluble polyphenol contents were observed in chapatis prepared from amylase-treated doughs compared with control chapatis. Extracts of chapatis prepared from amylase- and xylanase-treated doughs showed better antioxidant properties than extracts of control chapatis. Among these enzyme treatments, chapatis prepared from amylase-treated doughs showed better antioxidant properties than chapatis prepared from xylanase-treated doughs. High-performance liquid chromatography analysis of extracts of chapatis prepared from doughs treated with amylases showed the presence of potential antioxidant phenolic acids such as caffeic, gentisic and syringic acids in addition to the phenolic acids present in control chapatis. CONCLUSION: Treatment of doughs with amylases increased the contents of soluble dietary fibre and soluble polyphenols as well as improving the antioxidant properties of chapatis.

Beneficial effects of banana (Musa sp. var. elakki bale) flower and pseudostem on hyperglycemia and advanced glycation end-products (AGEs) in streptozotocin-induced diabetic rats

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Diabetes is a chronic health problem and major cause of death in most of the countries. Diet management plays an important role in controlling diabetes and its complications along with insulin and drugs. We have examined the effect of banana (Musa sp. var. elakki bale) flower and pseudostem on hyperglycemia and advanced glycation end-products (AGEs) in streptozotocin-induced diabetic rats. Our results indicated that banana flower and pseudostem have low glycemic index and have a high content of dietary fiber and antioxidants. Diabetic symptoms like hyperglycemia, polyuria, polyphagia, polydipsia, urine sugar, and body weight were ameliorated in banana flower- and pseudostem-treated rats. Increased glomerular filtration rate in the diabetic group (5.1 ± 0.22 ml/min) was decreased in banana flower-fed (2.5 ± 0.37 ml/min) and pseudostem-fed (3.0 ± 0.45 ml/min) groups and were significant at P < 0.001 and P < 0.01, respectively. Fructosamine and AGEs formed during diabetes were inhibited in treated groups when compared with the diabetic group. The diabetic group showed 11.5 ± 0.64 ìg of AGEs/mg protein in kidney, whereas, in banana flower- and pseudostem-fed groups, it was reduced to 9.21 ± 0.32 and 9.29 ± 0.24 ìg/mg protein, respectively, and were significant at P < 0.01. These findings suggest that banana flower and pseudostem have anti-diabetic and anti-AGEs properties and are beneficial as food supplements for diabetics (AU)

Beneficial effects of banana (Musa sp. var. elakki bale) flower and pseudostem on hyperglycemia and advanced glycation end-products (AGEs) in streptozotocin-induced diabetic rats.

Diabetes is a chronic health problem and major cause of death in most of the countries. Diet management plays an important role in controlling diabetes and its complications along with insulin and drugs. We have examined the effect of banana (Musa sp. var. elakki bale) flower and pseudostem on hyperglycemia and advanced glycation end-products (AGEs) in streptozotocin-induced diabetic rats. Our results indicated that banana flower and pseudostem have low glycemic index and have a high content of dietary fiber and antioxidants. Diabetic symptoms like hyperglycemia, polyuria, polyphagia, polydipsia, urine sugar, and body weight were ameliorated in banana flower- and pseudostem-treated rats. Increased glomerular filtration rate in the diabetic group (5.1 ± 0.22 ml/min) was decreased in banana flower-fed (2.5 ± 0.37 ml/min) and pseudostem-fed (3.0 ± 0.45 ml/min) groups and were significant at P < 0.001 and P < 0.01, respectively. Fructosamine and AGEs formed during diabetes were inhibited in treated groups when compared with the diabetic group. The diabetic group showed 11.5 ± 0.64 µg of AGEs/mg protein in kidney, whereas, in banana flower- and pseudostem-fed groups, it was reduced to 9.21 ± 0.32 and 9.29 ± 0.24 µg/mg protein, respectively, and were significant at P < 0.01. These findings suggest that banana flower and pseudostem have anti-diabetic and anti-AGEs properties and are beneficial as food supplements for diabetics.

Role of advanced glycation on aggregation and DNA binding properties of α-synuclein.

Parkinson's disease (PD) is a neurodegenerative disease with multiple etiologies. Advanced glycation end products (AGEs) accumulate in the aging brain and could be one of the reasons for age-related diseases like PD. Oxidative stress also leads to the formation of AGEs and may be involved in neurodegeneration by altering the properties of proteins. α-Synuclein is involved in pathogenesis of PD and there are limited studies on the role of AGE-α-synuclein in neurodegeneration. We studied the aggregation and DNA binding ability of AGE-α-synuclein in vitro. α-Synuclein is glycated using methylglyoxal and formation of AGE-α-synuclein is characterized using fluorescence studies, intrinsic tyrosine fluorescence, and fructosamine estimation. The results indicated that AGE-α-synuclein aggregates into smaller globular-like aggregates compared to fibrils formed with native α-synuclein. Further, it is found that AGE-α-synuclein induced conformational changes in scDNA from B-form to B-C-A mixed conformation. Additionally, AGE-α-synuclein altered DNA integrity as evidenced by the melting temperature, ethidium bromide, and DNAse I sensitivity studies. AGE-α-synuclein converted biphasic Tm to higher monophasic Tm. The Tm of AGE-α-synuclein-scDNA complex is more than that of native α-synuclein-scDNA complex, indicating that AGE-α-synuclein stabilized the uncoiled scDNA. AGE-α-synuclein could stabilize the uncoiled scDNA, as shown by the decrease in the number of ethidium bromide binding molecules per base pair of DNA. DNAse I sensitive studies indicated that both AGE-α-synuclein-scDNA and α-synuclein-scDNA are resistant to DNAse I digestion. The relevance of these findings to neuronal cell death is discussed.

Stimulation of glucose uptake by Musa sp. (cv. elakki bale) flower and pseudostem extracts in Ehrlich ascites tumor cells.

BACKGROUND: Glucose uptake study plays a major role in diabetes research. Impaired glucose uptake has been implicated in the development of hyperglycemia during diabetes. Banana plant is known for its anti-diabetic properties and our earlier report revealed that banana flower and pseudostem of Musa sp. cv. elakki bale is beneficial during diabetes in rat models. The present study was designed to evaluate the potential effect of banana flower and pseudostem extracts on glucose uptake in Ehrlich ascites tumor (EAT) cells using 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-D-glucose (2-NBDG), a fluorescent analogue of 2-deoxyglucose. RESULTS: Methanol and aqueous extracts of banana flower and pseudostem were more potent in promoting glucose uptake in EAT cells, in comparison to acetone and ethanol extracts. At 20 µg dosage, highest net glucose uptake was observed in aqueous extracts of banana flower (18.17 ± 0.43 nmol L⁻¹) and pseudostem (19.69 ± 0.41 nmol L⁻¹). Total polyphenol content was higher in methanol (9.031 ± 0.036 g kg⁻¹) and aqueous (6.862 ± 0.024 g kg⁻¹) extracts of banana flower compared to pseudostem, which were 0.442 ± 0.006 and 0.811 ± 0.011 g kg⁻¹, respectively. CONCLUSION: Banana flower and pseudostem extracts are able to promote glucose uptake into the cells, presumably through glucose transporters 1 and 3, which could be beneficial in diabetes. Glucose uptake is likely promoted by phenolic acids besides other bioactives. It can be hypothesized that consumption of nutraceutical-rich extract of banana flower and pseudostem could replace some amount of insulin being taken for diabetes.

Diabetes results in structural alteration of chondroitin sulfate/dermatan sulfate in the rat kidney: effects on the binding to extracellular matrix components.

Chondroitin sulfate (CS)/dermatan sulfate (DS) is a group of sulfated polymers, which play an essential role in various biological phenomena. In the kidney, they are present in small but significant amounts. Studies on their structure-function relationship in the kidney and their changes during diabetic conditions have not been rigorously looked into, which is the focus of this paper. The CS/DS content decreased significantly (14%) during diabetic conditions. This was accompanied by a decrease in the CS/heparan sulfate ratio. Disaccharide composition analysis revealed fine structural changes especially with respect to the E unit [glucuronic acid ß1-3 N-acetyl d-galactosamine (4,6-O-sulfate)] and the degree of sulfation. The mRNA expression levels of major enzymes involved in the synthesis of the "E"-disaccharide unit showed a decrease during diabetes. The changes in CS/DS had implications on ligand-binding properties when tested in vitro for binding to major extracellular matrix (ECM) components such as type IV collagen, laminin and fibronectin. Thus, this study provides insights into the structure-function relationship of CS/DS in the kidney during diabetes and alterations of which could aggravate conditions such as diabetic nephropathy by virtue of them being a part of ECM components.

Structural characterization of N-linked oligosaccharides of laminin from rat kidney: changes during diabetes and modulation by dietary fiber and butyric acid.

Carbohydrates of laminin, a family of large multidomain glycoproteins, have been implicated in various cellular activities including maintaining the protein structure, its function and also basement membrane integrity. During the course of our investigation, we observed that purified laminin from kidneys of control, diabetic, and dietary fiber- and butyric acid-treated diabetic rats showed differences in binding to extracellular matrix components. This prompted us to determine whether there are structural changes in laminin oligosaccharides. In this study, we have characterized a few major N-linked oligosaccharides isolated from purified laminin in various experimental groups, viz. normal, diabetic and diabetic rats fed with dietary fiber and butyric acid. Sugar composition, as identified by GLC, revealed the presence of mannose, galactose and N-acetylglucosamine. In order to study fine structures of the oligosaccharides, N-linked oligosaccharides of laminin were released by Peptide-N-glycosidase F digestion, end-labeled with 2-anthranilic acid and fractionated by lectin affinity chromatography. Furthermore, structural elucidation carried out by MALDI-TOF MS/MS analysis showed variations in the oligosaccharide sequence of laminin during diabetes which were altered by the feeding of dietary fiber and butyric acid.

Quercetin alleviates activities of intestinal and renal disaccharidases in streptozotocin-induced diabetic rats.

Specific activities of both intestinal and renal dissacharidases, such as sucrase, maltase, and lactase, were altered in diabetic rats. Our study was focused to evaluate the effect of feeding quercetin - a bioflavanoid on intestinal and renal dissacharidases in streptozotocin-induced diabetic rats. The rats were fed with 0.1% quercetin in diet. A reduction in intestinal maltase and sucrase, activities in quercetin-fed diabetic rats was observed in contrast to the increased activities in the starch-fed diabetic rats. A significant amelioration in renal dissacharidase activities in quercetin-fed diabetic rats was observed when compared to decreased activity in starch-fed diabetic rats.

Dietary fibres ameliorate decreased synthesis of heparan sulphate in streptozotocin induced diabetic rats.

The role of dietary fibers in diabetes has been studied by several workers. Long term dietary treatment with increased amounts of fiber-rich low-glycaemic index natural foods improves blood glucose and reduces the number of hypoglycemic events in type I diabetic patients. On the other hand Rohrbach and Martin and Cohen and Surma described changes in the general and biochemical structure of renal tissues such as the glomerular basement membranes. One of these changes was the reduction and undersulfation of the glycoconjugate and glycosaminoglycan heparan sulfate, which plays an important role in renal structure and function. The purpose of the present study was to determine specific effects of two types of dietary fiber on the composition of kidney glycoconjugates in an animal model of diabetes type I. Streptozotocin-treated diabetic rats were fed either a control diet or diets containing 10% wheat bran (insoluble dietary fiber) or 5% guar gum (soluble dietary fiber). Effects of these fibers on glycaemic control and nephropathy were assessed using previously described methodologies. The effect of dietary fiber in the glycoconjugate composition of kidneys of control and diabetic animals was studied by estimating their total hexose content, sulfated glycosaminoglycans, hexosamines and uronic acids. The activities of enzymes that participate in the synthesis of saccharides and glycoconjugates (L-glutamine-fructose-6-phosphate aminotransferase) and their degradation (N-acetyl-beta-glucosaminidase and beta-glucuronidase) were also evaluated. Results indicated that both soluble and insoluble dietary fibers ameliorated a significant increase in the activity of GFAT. Heparan sulfate was also isolated and quantified. Results indicated that the renal content of heparan sulfate decreased in diabetic animals and that this decrement was ameliorated by the ingestion of both soluble and insoluble fiber in the diet.

Structural features of arabinoxylans from bajra (pearl millet).

Structures of arabinoxylans (AX) from bajra, from a variety known to have characteristic quality in the making of roti, an unleavened flat bread, were elucidated by a combination of methylation analysis, (13)C NMR, FT-IR, etc. Arabinoxylans isolated from barium hydroxide-extracted polysaccharides and hemicellulose B had a backbone of xylose residues with beta-(1,4) linkages and were branched mainly through alpha-(1,3) linkages by arabinofuranosyl residues. Completely branched xylose residues were also present. The AX from bajra characteristically had large numbers of branches, and this may be one of the reasons for the crispiness of the rotis.