Background: MicroRNA-122-5p (miR-122), a liver-specific miR, levels are increased in the blood during obesity and diabetes and is taken-up by the extra-hepatic tissues including vascular endothelium. Despite miR-122’s role in maintaining hepatic homeostasis, its uptake (from circulation) by other tissues disrupts their function and increases the risk of cardiovascular disorders. Hence, inhibiting extra-hepatic miR-122 is a viable approach to mitigate vascular disorders associated with diabetes. Hypothesis: Endothelium-targeted miR-122 inhibitor selectively targets miR-122 in endothelial cells and improves vascular endothelial function in high-fat-diet (HFD) fed pre-diabetic mice. Methods: To selectively inhibit miR-122 in the endothelial cells, we synthesized gamma-peptide nucleic acid miR-122 inhibitor (γP-122 I) tagged with vascular cell adhesion molecule 1 (VCAM1) targeted peptide (e-γP-122 I). The purity was assessed using HPLC. Pre-diabetic condition was induced by feeding mice with a HFD (for 8 weeks) and e-γP-122-I was injected to the HFD-fed mice (2 weeks after dietary intervention; 5 mg/kg/day, i.p., 6 weeks). MiR-122 expression was measured by real-time PCR. The vascular endothelial function was assessed by determining endothelium-dependent and independent relaxation of phenylephrine-induced precontracted aortic rings. Results were expressed as mean ± s.e.m. Results: The e-γP-122-I was >95% pure. The HFD-fed mice show higher levels of serum and aortic miR-122 expression and impaired vascular endothelial function. e-γP-122-I treatment selectively inhibited aortic miR-122 expression while it has no effect on other organs (heart, kidney etc.) and improved vascular endothelial function and inflammation. Conclusions: e-γP-122-I selectively inhibits miR-122 in the endothelium and improves endothelial function in pre-diabetic mice.

Cardiotoxicity caused by adrenergic receptor agonists overdosing or stress-induced catecholamine release promotes cardiomyopathy, resembling Takotsubo cardiomyopathy (TC). TC is characterized by transient regional systolic dysfunction of the left ventricle. The animal models of TC and modalities for assessing regional wall motion abnormalities in animal models are lacking. We previously reported the protective role of a small noncoding microRNA-204-5p (miR-204) in cardiomyopathies, but its role in TC remains unknown. Here we compared the impact of miR-204 absence on phenylephrine (PE)-induced and transaortic constriction (TAC)-induced changes in cardiac muscle motion in the posterior and anterior apical, mid, and basal segments of the left ventricle using 2-dimensional speckle-tracking echocardiography (2-STE). Wildtype and miR-204-/- mice were subjected to cardiac stress in the form of PE for four weeks or TAC-induced pressure overload for five weeks. PE treatment increased longitudinal and radial motion in the apex of the left ventricle and shortened the peak motion time of all left ventricle segments. The TAC led to decreased longitudinal and radial motion in the left ventricle segments, and there was no difference in the peak motion time. Compared to wildtype mice, PE-induced peak cardiac muscle motion time in the anterior base of the left ventricle was significantly earlier in the miR-204-/- mice. There was no difference in TAC-induced peak cardiac muscle motion time between wildtype and miR-204-/- mice. Our findings demonstrate that PE and TAC induce regional wall motion abnormalities that 2-STE can detect. It also highlights the role of miR-204 in regulating cardiac muscle motion during catecholamine-induced cardiotoxicity.

This study identifies the anti-inflammatory, antioxidant, and immunomodulatory potential of a fatty acid methyl ester segregated from the brown algae Turbinaria ornata and identified by nuclear magnetic resonance and mass spectrometry as methyl 6,12-dimethyltridecanoate (ET). Antioxidant and anti-inflammatory effects of ET were studied on lipopolysaccharide (LPS)-induced inflammatory reaction in RAW 264.7 macrophages. Moreover, in silico docking studies of isolated ET with inflammatory markers TNFα, NFκB, and COX-2 showed potent binding scores suggesting anti-inflammatory potential. ET significantly reduced LPO and increased LPS-induced SOD, catalase, and GSH levels. Molecular docking results were further confirmed by checking mRNA levels of selected cytokines (IL6 and IL10), followed by protein expression of iNOS and NFκB in LPS-induced macrophages. ET significantly upregulated the expression of IL10 and downregulated the expression of IL6, iNOS, and NFκB, confirming the inhibition of LPS-induced inflammation via the iNOS/NFκB pathway.

The new mixed-ligand copper(II) complexes [Cu(FAA)(diimine)(H2O)](ClO4)(2) (1-4) [FAA is folic acid-conjugated ethyl pyridyl amine; diimine is 1,10-phenanthroline (phen; 1), 5,6-dimethyl-1,10-phenanthroline (5,6-dmp; 2), 3,4,7,8-tetramethyl-1,10-phenanthroline (3,4,7,8-tmp; 3), and dipyrido-[3,2-f:2',3'-h]-quinoxaline (dpq; 4)] were synthesized and characterized adopting appropriate analytical and spectroscopic techniques. DNA and protein binding studies showed that the complex [Cu(FAA)(5,6-dmp)(H2O)](2+) (2) has higher DNA- and protein-binding abilities than the other complexes. In the presence of the activating agent ascorbic acid, the same complex 2 caused pronounced DNA cleavage. The mechanistic study revealed that the oxidative DNA cleavage induced by complex 2 was effected through the freely diffusible hydroxyl radical; the same complex induced the highest incidence of cytotoxicity in the triple-negative breast cancer cell MDA-MB-231 (IC50) = 0.54 mu M) compared to its congeners in the series, cisplatin, and the free ligand (FAA). AO/EthBr staining assay revealed induction of cell death by apoptosis. Interestingly, the complex 2 demonstrated greater selectivity toward cancer cells as revealed in only the negligible cytotoxicity to normal cells (lymphocytes), which emphasizes an important role to the cancer-targeting ligand folic acid conjugated to the Cu(II) complexes in dealing with triple-negative breast cancers.

Introduction and Aim: Hyperglycemia is the signature style of type II diabetes (T2D) which impairs pathways of insulin signaling and endoplasmic reticulum (ER) stress. Diosgenin is a popular saponin known for its antidiabetic and anti-obesity properties predominantly found in Dioscorea species. However, the mechanism of hypoglycemic effects related to pathways of ER stress, insulin signaling, and uptake of glucose in the liver gained less attention. Accordingly, this study was aimed to assess the effect of Diosgenin on the uptake of glucose, insulin signaling as well as on ER stress in human HepG2 cells cultured in hyperglycemic or high glucose (HG) condition. Materials and Methods: Viability studies on HepG2 cells, assay for studying the uptake and consumption of glucose, gene expression, and docking studies were carried out. Results: The treatment with Diosgenin was found to elevate the uptake as well as consumption of glucose in HepG2 cells under hyperglycemic conditions. Diosgenin prevented inactivation of the PI3K Akt pathway as well as GLUT4 levels induced by high glucose. Furthermore, endoplasmic reticulum stress elements viz., inositol requiring enzyme 1 (IRE1), protein kinase like endoplasmic reticulum kinase (PERK), C/EBP homologous protein (CHOP) and sXBP1 were reduced by diosgenin treatment provoked by high glucose returning to normal ER homeostasis. Moreover, the effects are supported by the binding efficiency of diosgenin with insulin signaling proteins using bioinformatic tools. Conclusion: Our findings suggest that diosgenin elevated the uptake of glucose by hindering the inactivation of PI3K Akt GLUT4 pathway and ER stress caused by high glucose, thus, preventing further hepatic dysfunction.

The influenza A virus (IAV) is a highly contagious virus that causes pandemics and seasonal epidemics, which are major public health issues. Current anti-influenza therapeutics are limited partly due to the continuous emergence of drug-resistant IAV strains; thus, there is an unmet need to develop novel anti-influenza therapies. Here, we present a novel imidazo[1,2-a]pyrimidine scaffold that targets group 2 IAV entry. We have explored three different regions of the lead compound, and we have developed a series of small molecules that have nanomolar activity against oseltamivir-sensitive and -resistant forms of group 2 IAVs. These small molecules target hemagglutinin (HA), which mediates the viral entry process. Mapping a known small-molecule-binding cavity of the HA structure with resistant mutants suggests that these molecules bind to that cavity and block HA-mediated membrane fusion.

A series of mixed ligand copper(II) complexes, formulated as [Cu(L1-L5)(phen)(H2O)](ClO4)(2) (1-5), where phen = 1,10-phenanthroline, L1 = 2-pyridin-2-yl-quinoline, L2 = 2-pyridin-2-yl-quinoxaline, L3 = 6,7-dimethyl-2-pyridin-2-yl-quinoxaline, L4 = 4-phenyl-2-pyridin-2-yl-quinoline, and L5 = 4-phenyl-2-pyridin-2-yl-quinazoline, were synthesized and characterized. The molecular structure of 3, which alone formed into appreciable crystals, was determined by single-crystal X-ray studies, and the coordination geometry around Cu(II) was nearly square pyramidal (tau, 0.092). DNA and protein binding, DNA cleavage and in vitro cytotoxicity of the mixed ligand complexes 1-5 were investigated and compared with their analogue bis-complexes [Cu(L1-L5)(2)H2O](ClO4)(2) 6-10. All five mixed ligand complexes exhibited efficient DNA and protein binding, wherein 5 was the most potent. DNA cleavage studies revealed that all mixed ligand complexes engage in self-activated DNA cleavage, with 2 producing full conversion of supercoiled DNA to nicked circular form. Complex 5, with the highest DNA- and protein-binding efficiencies, demonstrated the highest cytotoxicity to A549 non-small human lung carcinoma cell (IC50 = 3.85 lM), three times more potent than cisplatin. Metal-assisted reactive oxygen species (ROS) were found to be responsible for cytotoxicity of the complexes. Fluorescent staining assays showed that all complexes induce apoptotic cell death along with some degree of necrosis. Western blot analysis of caspase-3 expression of cells exposed to Cu(II) complexes 1 and 5 revealed that both promote apoptosis, with 5 demonstrating more potency. Thus, the mixed ligand copper complexes demonstrated efficient biological activity compared to bis-complexes, with 5 holding promise for future investigation towards development as a cancer therapeutic. (C) 2020 Elsevier Ltd. All rights reserved.

Background: Dyslipidemia a common pathology in metabolic syndrome results due to various factors. Although statins are proven for its beneficial role in the management of hyperlipidemia, plant-based foods and their ingredients are still a cornerstone of health care and medical prescriptions. Legumes are important constituents of a balanced diet. They are rich in fiber and provide health benefits and are used to treat disease related pathologies. Purpose: The present study intended to identify the influence of two legumes, horsegram and groundnut individually and with atorvastatin in high fat diet induced hyperlipidemic rats and trace if there is any adverse effect due to food drug interactions. Methods: Hyperlipidemia in Sprague Dawley rats were induced by feeding high fat diet for 12 weeks. Subsequently, supplementation of horsegram and groundnut with and without atorvastatin was given for 4 weeks. The physiological indicators (body weight, feed intake, fat index), biochemical parameters (cholesterol, triglyceride, glucose, urea and creatinine) including markers of oxidative stress (catalase, GST, SOD, TBARS and GSH), mRNA expression markers of lipid metabolism and inflammation markers (SREBP-1, ACC, FABP, HMG-CoAR, NF-kB and IL-6) and the protein expression of HO-1 and NF-kB followed by histopathological analysis of the heart, liver and kidney tissues were checked to study the influence of the legumes with and without atrovastatin. Results: Results showed that administration of the legumes individually and their combination with atorvastatin significantly reduced the body weight, feed intake, fat index, biochemical metabolites and enzymatic markers in comparison to the HFD group. In addition, it also decreased the enzymatic and non-enzymatic antioxidant levels. Likewise, the legumes individually as well as in combination with atorvastatin significantly decreased the protein expression of HO-1 and NF-kB. Further, histopathological analysis confirmed the influence of the legumes and its combination to overcome hyperlipidemia as evidenced by the pathological changes in the treated rats. Conclusion: The results of the present work confirm that the legumes individually as well as its combination with atorvastatin are helpful to manage hyperlipidemia. On comparison, horsegram with atorvastatin possess a better therapeutic efficacy when compared to groundnut with atorvastatin and there is no evidence of food drug interactions.

It is well documented that influenza A viruses selectively package 8 distinct viral ribonucleoprotein complexes (vRNPs) into each virion; however, the role of host factors in genome assembly is not completely understood. To evaluate the significance of cellular factors in genome assembly, we generated a reporter virus carrying a tetracysteine tag in the NP gene (NP-Tc virus) and assessed the dynamics of vRNP localization with cellular components by fluorescence microscopy. At early time points, vRNP complexes were preferentially exported to the MTOC; subsequently, vRNPs associated on vesicles positive for cellular factor Rab11a and formed distinct vRNP bundles that trafficked to the plasma membrane on microtubule networks. In Rab11a deficient cells, however, vRNP bundles were smaller in the cytoplasm with less co-localization between different vRNP segments. Furthermore, Rab11a deficiency increased the production of non-infectious particles with higher RNA copy number to PFU ratios, indicative of defects in specific genome assembly. These results indicate that Rab11a+ vesicles serve as hubs for the congregation of vRNP complexes and enable specific genome assembly through vRNP:vRNP interactions, revealing the importance of Rab11a as a critical host factor for influenza A virus genome assembly.

Background: Inflammation and oxidative stress are common pathologies in a wide range of chronic diseases. Polysaccharides are known to exhibit antioxidant and anti-inflammatory potential and are suggested to possess immunomodulatory potential. Purpose: Herein, the immunomodulatory activity of a sulfated polysaccharide (PS) separated from a brown marine algae Turbinaria ornata is studied in LPS instigated systemic inflammation in experimental rats. Study design and methods: Male SD rats are pretreated with different doses of PS (2.5, 5, 10 mg/kg bw) for a week followed by inducing systemic inflammation using LPS (10 mg/kg i.p.). Blood withdrawn after 8 h of LPS injection is subjected to hematological analysis (WBC, HCT, and PLT). After 24 h of LPS induction, cardiac tissue was isolated and subjected to biochemical, molecular, and histopathological analysis. Effect of PS pre-treatment (2.5, 5, 10 mg/kg bw) was checked by assessing serum parameters (AST, CK-MB, and gamma GT), antioxidant markers (LPO, GSH, SOD, Grx) and inflammatory markers (IL1 beta, IL6, IL10, NF kappa B), followed by analyzing the iNOS, PI3k and Akt to identify the probable mode of action. Results: Elevated levels of AST, CK-MB, and gamma GT in serum were significantly reduced on PS pretreatment. LPS significantly raised the LPO and Grx levels in heart tissue whereas, PS pre-treatment significantly reduced LPO and Grx levels. GSH and SOD levels were reduced upon LPS induction and were brought to near normal by HD of PS. PS also reduced the mRNA levels of IL6, Trx, and increased IL10 levels in the heart tissue substantiating its anti-inflammatory and antioxidant potency. Further, IL1 beta, NF kappa B, iNOS, and pPI3k/pAkt expressions were significantly modulated by PS in the cardiac tissue substantiating the immunomodulatory effect. A trend of improvement in the inflammatory pathology was also observed in the heart tissue compared to LPS control, as confirmed by histopathology analysis. Conclusion: Altogether, this study concludes the immunomodulatory potential of PS from the marine macroalgae Turbinaria ornata significantly and prevents LPS induced systemic inflammation in the cardiac tissue presumably influenced by the glucopyranose and fucopyranose subunits in the polysaccharide.

Background: Inflammation and pain, mainly induced by the prostaglandins synthesized by the cyclooxygenase enzymes, may cause distress. To overcome this unpleasant stress in a safer manner, numerous natural molecules are proven for modulating the COX enzymes. Epicatechin and daidzein are two bioactive natural compounds present in horsegram, a legume known for its medicinal properties. Objective: The present study aims at evaluating the potential of horsegram. and some of its bioactive molecules, to be used as an anti-inflammatory and analgesic agent mediated by the inhibition of COX enzymes, which can be recommended as a substitute for chemically synthesized NSAIDs. Methods: The present work involved the quantification of epicatechin and daidzein present in horsegram seeds. The COX enzyme inhibitory nature of epicatechin and daidzein was tested using in silico docking analysis with Autodock software and was further confirmed by in vitro COX inhibitory biochemical assays. Furthermore, the anti-inflammatory and analgesic activities of the horsegram seeds were evaluated in animal experiments. Results: Horsegram seeds contain 158.1 microgram/g and 6.51 microgram/g of epicatechin and daidzein respectively. The docking studies reveal that both the bioactive molecules exhibit better binding efficiency with COX-2 protein as compared to COX-1. Hence, in vitro COX-2 inhibitory assay was performed for epicatechin, daidzein and compared with known analgesic agent diclofenac which revealed a pronounced dose dependent inhibitory activity. Furthermore, the analgesic and anti-inflammatory activity of horsegram in experimental animals exhibited a dose dependent effect which might be due to the presence of the bioactive compounds such as epicatechin and daidzein. Conclusion: The results suggest that epicatechin and daidzein present in horsegram are potent cyclooxygenase inhibitors and thus would be helpful in the management of inflammation and pain.

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A sedentary lifestyle combined with the intake of high-calorie diet has been the paramount cause of metabolic syndrome (MS) which is now a serious concern of public health worldwide as it involves the coexistence of hypertension, hyperlipidemia, glucose intolerance, and obesity. Hence, identifying a suitable strategy to overcome the worldwide menace of MS is imperative. Macrotyloma uniflorum a lesser known legume is highly nutritious and notable for its ethano-medicinal potential. Herein, the influence of M. uniflorum in high-fat dietinduced metabolic changes in a rodent model of metabolic syndrome was evaluated. Serum levels of glucose, total cholesterol, triglycerides, VLDL-c, and bodyweight were decreased, whereas HDL-c was increased in M. uniflorum-treated MS rats. The protein expression (AMPK-α, PPAR-α, and PPAR-γ) and gene expression (leptin, adiponectin, resistin, UCP2, NF-κB, and IL-6) results are impressive to highlight that M. uniflorum modulates the pathological conditions of MS and proves to be cardioprotective. Furthermore, the histopathological analysis confirmed the pathological changes and substantiates the influence of M. uniflorum to overcome MS. The HPLC and GC (MS) profiling reveals the presence of an array of polyphenols such as rutin (694.61 μg/g), catechin (500.12 μg/g), epicatechin (158.10 μg/g), gallic acid (17.98 μg/g), ferulic acid (10.911 μg/g), daidzein (6.51 μg/g), and PUFA, respectively, which probably exhibits the therapeutic effect on MS and associated complications by modulating lipid metabolism and adipogenesis. PRACTICAL APPLICATIONS: Metabolic disorders like CVD and diabetes are leading cause of mortality and morbidity worldwide. With emerging issues on adverse effects of modern drugs, the emphasis on "Food is Medicine and Medicine as Food" has taken dramatic dimensions in the healthcare sector. Therefore, nutraceuticals are in great demand in the developed world off late. Legumes, are potent elements in a balanced diet next to cereals. Exploring the medicinal properties of legumes could bring a revolution in public health and nutraceutical industries. This study scientifically validated the phytochemicals in M. uniflorum for its functional potential in the management of Metabolic Syndrome (MS). This study would help the nutraceutical industries to develop functional foods using M. uniflorum seeds to make porridges and soups or nutraceutical supplements with the bioflavonoids isolated from M. uniflorum for the management of metabolic disorders by mitigating hyperlipidemia, oxidative stress, and inflammation.

Background: Marine macroalgae known for its polysaccharides exhibit potent biomedical properties and its potential as an anti-inflammatory agent has increased in the recent past as inflammation is a major pathology noted in many chronic diseases. Purpose: The present study investigates the anti-inflammatory potential of a sulfated polysaccharide (PS) isolated from the marine algae Turbinaria ornata collected from the Indian waters on LPS induced inflammation in RAW 264.7 macrophages. Study design and methods: PS isolated from the macroalgae was characterized using ESI(MS) and was screened for its antioxidant and anti-inflammatory potential in RAW 264.7 cells by assessing markers of oxidative stress, and inflammation. Results: LPS significantly increased the levels of LPO and LDH in RAW 264.7 cells which were significantly reduced in PS pre-treatment groups. Pretreatment significantly increased the antioxidants GSH and SOD and significantly reduced mRNA levels of IL6 and TNF alpha in vitro confirming its anti-inflammatory potential. NF kappa B and iNOS were significantly modulated by PS confirming the probable mode of action. Conclusion: Altogether, it can be concluded that PS isolated from Turbinaria ornata collected from the Southeast Coast of India exhibits antioxidant and anti-inflammatory potential probably mediated by the sulfated polysaccharide containing glucopyranose and fucopyranose moieties. (C) 2020 Elsevier B.V. All rights reserved.

Cholest-4-ene-3,6-dione (KS) is a cholesterol oxidation product which exhibits anti-proliferative activity. However, its precise mechanism of action remains unknown. In this study, the effects of KS on AKR1C3 inhibition and anti-proliferative activities were investigated in the hormone-dependent MCF-7 breast cancer cells. We identified that KS arrested the enzymatic conversion of estrone to 17-β estradiol, by inhibiting AKR1C3 in intact MCF-7 cells. The anti-proliferative effects of KS were evaluated by MTT assay, acridine orange and ethidium bromide dual staining, cell cycle analysis and Western blotting. KS arrested the cell cycle progression in the G1 phase with a concomitant increase of the Sub-G0 population to increase in concentration and time. It also enhanced the p53 and NFkB expression and induced caspase-12, 9 and 3 processing and down-regulated the Bcl-2 expression. Molecular docking studies performed to understand the inhibition mechanism of KS on AKR1C3 revealed that KS occupied the binding region of AKR1C3 with almost similar orientation as indomethacin (IM), thereby acting as an antagonistic agent for AKR1C3. Based on the results it is identified that KS induces inhibition of AKR1C3 and cell death in MCF-7 cells. These results indicate that KS can be used as a molecular scaffold for further development of novel small-molecules with better specificity towards AKR1C3.

BackgroundCardiovascular disease and its related deaths are increasing in the modern world. Therefore, there is a need to identify a plant based nutraceutical supplement with potent activity.Hypothesis/PurposeReportedly, the protective effect of the rutin in hypoxia-induced cardiomyocytes is due to the activation of molecular networks related to programmed cell death.Study Design-MethodsPhytochemical methods and advanced analytical methods were employed to isolate natural products from Spermococe hispida their effects in cardiomyocyets.ResultsWe reports herein that CoCl2-induced hypoxic condition significantly decreased cell viability as evidenced by MTT assay and cell cycle analysis. Western blot studies revealed an up-regulation of HIF-1α, BAX and caspase and down-regulation of BCl-2 expression, followed by modulation of Akt, p-Akt, p38 and p-p38. The oxidative abnormalities were ameliorated by rutin pretreatment, as deduced by the reduced CoCl2-induced cytotoxicity, MDA concentration and LDH activity and the enhanced levels of GSH and SOD in a dose-dependent manner. Rutin protects H9c2 cells from CoCl2-induced hypoxic damage by mitigating oxidative stress and preserving cell viability by modulating the antiapoptotic proteins.ConclusionThe overall findings reinforce the cardioprotective action of rutin, a potential source of antioxidant of natural origin, which may help in mitigating the progress of oxidative stress in hypoxic conditions such as myocardial infarction and stroke.