Gene expression profile analysis unravelled the systems level association of renal cell carcinoma with diabetic nephropathy and Matrix-metalloproteinase-9 as a potential therapeutic target.

Type 2 diabetes (T2D) and cancer share many common risk factors. However, the potential biological link that connects the two at the molecular level is still unclear. The experimental evidence suggests that several genes and their pathways may be involved in developing cancerous conditions associated with diabetes. In this study, we identified the protein-protein interaction (PPI) networks and the hub protein(s) that interlink T2D and cancer using genome-scale differential gene expression profiles. Further, the PPI network of AMP-activated protein kinase (AMPK) in cancer was analyzed to explore novel insights into the molecular association between the two conditions. The densely connected regions were analyzed by constructing the backbone and subnetworks with key nodes and shortest pathways, respectively. The PPI network studies identified Matrix-metalloproteinase-9 (MMP-9) as a hub protein playing a vital role in glomerulonephritis tubular diseases and some genetic kidney diseases. MMP-9 was also associated with different growth factors, like tumor necrosis factor (TNF-α), transforming growth factor 1 (TGF-1), and pathways like chemokine signaling, NOD-like receptor signaling, etc. Further, the molecular docking and molecular dynamic simulation studies supported the druggability of MMP-9, suggesting it as a potential therapeutic target in treating renal cell carcinoma linked with diabetic kidney disease.Communicated by Ramaswamy H. Sarma.

Evaluation of diosgenin content from eleven different Indian varieties of fenugreek and fenugreek leaf powder fortified bread.

The present study designed to establish the diosgenin profile from eleven different Indian varieties of fenugreek, and subsequently develop powder from the best stage and variety to prepare diosgenin fortified bread. The seeds, sprouts and leaves of different fenugreek varieties were analyzed for the diosgenin using HPLC. The content of phenolics and flavonoids also estimated and assessed for its antioxidant capacity using phosphomolybdate, DPPH and FRAP. Among eleven varieties screened for diosgenin and the other compounds, tender leaf of Kasuri methi (KS) variety showed the highest content of diosgenin (466.89 ± 0.32 mg/100 g FW), phenolics (58.54 ± 2.70 mg/100 g FW) and flavonoids (1104.16 ± 43.70 mg/100 g FW) followed by Pusa Early Bunching (PEB) (444.18 ± 0.36 mg/100 g FW) and Early Bunching (EB) (409.45 ± 0.42 mg/100 g FW). Among three stages, seeds found to be a better total antioxidant, DPPH scavenging, and reducing ability. Further, based on the results, bread fortified with 1.5% KS leaf powder is found to be optimal which also has significant diosgenin content (268.91 mg/100 g DW). And the effect of KS leaf powder on amylograph and farinograph characteristics of wheat flour and quality characteristics of bread showed promising results of acceptance.

Atheroprotective effect of novel peptides from Porphyridium purpureum in RAW 264.7 macrophage cell line and its molecular docking study.

OBJECTIVE: To explore the atherogenic foam cell prevention efficiency of two dipeptides purified from Porphyridium purpureum on RAW 264.7 cell line and to study its molecular interaction through molecular docking. RESULT: P. purpureum consists of 29.9% protein and 2.98% phycoerythrin on a dry weight basis. The two dipeptides namely of Histidine-Glutamic acid (HE) and Glycine-Proline (GP) isolated from the total protein and purified phycoerythrin of P. purpureum respectively, were evaluated for atherogenic foam cell prevention capacity in RAW 264.7 cell line. The IC5O values of peptides were found to be 91.2 ± 1.81 µg/ml (GP), 103.3 ± 4.8 µg/ml (HE) in MTT assay. The two peptides reduce the foam cell formation, intracellular lipid accumulation (cholesterol and triglycerides) and the secretion of TNF-α and IL-6 which are inflammatory cytokines in RAW 264.7 cell line at non-cytotoxic concentrations. A molecular interaction study proposed the binding pose for GP and HE peptides targeting the scavenging receptors CD36, SRA1, and Map Kinase p38 (a protein mediator). CONCLUSIONS: The cell line and molecular docking study indicated that among the two dipeptides, peptide GP has the highest atherogenic foam cell prevention efficiency.

Differential expression of anthocyanin biosynthesis genes in Daucus carota callus culture in response to ammonium and potassium nitrate ratio in the culture medium.

Anthocyanins are major water-soluble and dynamic colouring plant pigment present in plant tissues with the high antioxidant properties. The role of ammonium and potassium nitrate in the culture medium on anthocyanin augmentation is probed thoroughly, but the mechanism of its biosynthesis continues to be unclear. Hence, the present study was undertaken to optimise nitrate ratio in the culture medium for anthocyanin augmentation and examination of its biosynthesis pathway in callus culture of Daucus carota. MS basal medium fortified with various ratio of NH4NO3:KNO3 was employed to find their impact on biomass, anthocyanin augmentation and the expression profile of anthocyanin biosynthesis genes in the callus culture. The data indicated that the highest anthocyanin content (9.30 ± 0.25 mg/100 g FW) was seen in callus grown on the medium supplemented with 20.0 mM NH4NO3:37.6 mM KNO3 and the least was seen in the medium which contained 40.0 mM NH4NO3:18.8 mM KNO3 (2.74 ± 0.27 mg/100 g FW). This indicates an optimal concentration of NH4NO3:KNO3 ratio is essential to produce a higher amount of anthocyanin in in vitro culture. Meanwhile, anthocyanin biosynthesis genes were differentially expressed as confirmed by qRT-PCR in the time interval of 5, 10, 15, 20 and 25 days. The transcript levels of nine anthocyanin biosynthesis genes were increased in the response of varying NH4NO3:KNO3 ratio in the medium. The transcript level of early genes PAL, 4CL, CHS and CHI increased by 19.5, 21.0, 16.2 and 9.98-fold, respectively, compared with control. In addition, late biosynthesis genes LDOX and UFGT resulted in the transcript level of 11.3 and 13.6-fold, respectively.

Fermentation induced changes in bioactive properties of wine from Phyllanthus with respect to atherosclerosis.

Wine was prepared from three varieties of Phyllanthus viz., P. emblica (wild and cultivated) and P. acidus. Among the wines prepared, cultivated Emblica wine had the highest total phenolic (11.02 µg gallic acid equivalent/ml) and flavonoid (59.46 µg quercetin equivalent/ml) content. Further gallic acid, syringic acid, protocatechuic acid and caffeic acid were present in higher amounts in wine from the cultivated variety compared to other wines. HPLC analysis showed that in juice of the cultivated variety, gallic acid and coumaric acid were found in higher amounts than that in the corresponding wine. Antioxidant assays, LDL oxidation prevention, foam cell prevention and nitrite scavenging activities (cell lines) were found to be highest in cultivated Emblica juice and wine with an activity of 15 µg/ml and 14 µg/ml (nitrite assay) and 108.649 µg ascorbic acid equivalent/mg and 321.622 µg ascorbic acid equivalent /mg (total antioxidant capacity) respectively. CD36 expression was reduced and ABCA1 expression was increased to the highest extent by the cultivated Emblica wine and juice. Further, antioxidant activity was seen to increase during the course of fermentation. Sensory analysis showed that cultivated Emblica wine was sweeter compared to the other wines.

Inhibition of LDL oxidation and oxidized LDL-induced foam cell formation in RAW 264.7 cells show anti-atherogenic properties of a foliar methanol extract of Scoparia dulcis.

BACKGROUND: Oxidation of low density lipoproteins and their further uptake by macrophages is known to result in the formation of foam cells, which are critical in the initiation of atherosclerosis through activation of inflammatory signalling cascades. Thus, powerful dietary antioxidants are receiving attention for the reversal of such pathological states. MATERIALS AND METHODS: Extracts of Scoparia dulcis have been used as tea and health drinks with various health promoting effects. In the present study, we examined the reactive oxygen scavenging potential as well as anti-inflammatory and anti-atherogenic efficacies, using leaf extracts obtained after successive extraction with various solvents. RESULTS: A methanol extract showed potent antioxidant activity with an IC50 value of 570 µg/ml, caused hydrogen peroxide scavenging (28.9 µg/ml) and anti-inflammatory effects by improving human erythrocyte membrane stabilisation (about 86%). The methanol extract also efficiently inhibited lipid peroxidation and oxidation of low density lipoproteins, thus preventing foam cell formation in cultured RAW 264.7 cells. Furthermore, phytochemical screening of the extracts showed high accumulation of flavonoids. CONCLUSIONS: The foliar methanol extract of Scoparia dulcis has a strong anti-atherogenic potential and this property could be attributed maybe due to presence of flavonoids since HPLC analysis showed high concentrations of myricetin and rutin in the methanol extract.

Silicon-induced changes in antifungal phenolic acids, flavonoids, and key phenylpropanoid pathway genes during the interaction between miniature roses and the biotrophic pathogen Podosphaera pannosa.

Application of 3.6 mm silicon (Si+) to the rose (Rosa hybrida) cultivar Smart increased the concentration of antimicrobial phenolic acids and flavonoids in response to infection by rose powdery mildew (Podosphaera pannosa). Simultaneously, the expression of genes coding for key enzymes in the phenylpropanoid pathway (phenylalanine ammonia lyase, cinnamyl alcohol dehydrogenase, and chalcone synthase) was up-regulated. The increase in phenolic compounds correlated with a 46% reduction in disease severity compared with inoculated leaves without Si application (Si-). Furthermore, Si application without pathogen inoculation induced gene expression and primed the accumulation of several phenolics compared with the uninoculated Si- control. Chlorogenic acid was the phenolic acid detected in the highest concentration, with an increase of more than 80% in Si+ inoculated compared with Si- uninoculated plants. Among the quantified flavonoids, rutin and quercitrin were detected in the highest concentrations, and the rutin concentration increased more than 20-fold in Si+ inoculated compared with Si- uninoculated plants. Both rutin and chlorogenic acid had antimicrobial effects on P. pannosa, evidenced by reduced conidial germination and appressorium formation of the pathogen, both after spray application and infiltration into leaves. The application of rutin and chlorogenic acid reduced powdery mildew severity by 40% to 50%, and observation of an effect after leaf infiltration indicated that these two phenolics can be transported to the epidermal surface. In conclusion, we provide evidence that Si plays an active role in disease reduction in rose by inducing the production of antifungal phenolic metabolites as a response to powdery mildew infection.