Role of miR-204 in segmental cardiac effects of phenylephrine and pressure overload.

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.

Methyldecanoate isolated from marine algae Turbinaria ornata enhances immunomodulation in LPS-induced inflammatory reactions in RAW 264.7 macrophages via iNOS/NFκB pathway.

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.

Synthesis, Optimization, and Structure-Activity Relationships of Imidazo[1,2-a]pyrimidines as Inhibitors of Group 2 Influenza A Viruses.

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.

Immunomodulatory activity of brown algae Turbinaria ornata derived sulfated polysaccharide on LPS induced systemic inflammation.

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 γGT), antioxidant markers (LPO, GSH, SOD, Grx) and inflammatory markers (IL1ß, IL6, IL10, NFκB), followed by analyzing the iNOS, PI3k and Akt to identify the probable mode of action. RESULTS: Elevated levels of AST, CK-MB, and γGT 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, IL1ß, NFκB, 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.

Host factor Rab11a is critical for efficient assembly of influenza A virus genomic segments.

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.

Sulfated polysaccharide from Turbinaria ornata suppress lipopolysaccharide-induced inflammatory response in RAW 264.7 macrophages.

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 TNFα in vitro confirming its anti-inflammatory potential. NFκB 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.

Type 5 17-hydroxysteroid dehydrogenase/prostaglandin F synthase (AKR1C3) inhibition and potential anti-proliferative activity of cholest-4-ene-3,6-dione in MCF-7 breast cancer cells.

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-ß 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.

Correction to: Neophytadiene from Turbinaria ornata Suppresses LPS-Induced Inflammatory Response in RAW 264.7 Macrophages and Sprague Dawley Rats.

The publisher made a mistake in the published version of this article.

Neophytadiene from Turbinaria ornata Suppresses LPS-Induced Inflammatory Response in RAW 264.7 Macrophages and Sprague Dawley Rats.

This study investigates the mode of action of Neophytadiene (MT), a molecule isolated from a marine algae Turbinaria ornata in LPS-induced inflammation in both in vitro and in vivo conditions. Neophytadiene (25, 50, 100 µM/mL) was treated to LPS-stimulated RAW 264.7 macrophages cells to identify its anti-inflammatory potential by measuring the level of tumour necrosis factor (TNF-α) by enzyme-linked immunosorbent assay (ELISA) and nitric oxide (NO) using Griess reagent. The mRNA levels of inflammatory cytokines, interleukin (IL-6 and IL-10), and the protein expression of nuclear factor-κB (NF-κB) and inducible nitric oxide synthase (iNOS) were quantified by Western blot analysis. Subsequently, Neophytadiene (12, 25, 50 mg/kg b.wt/p.o) was pre-treated for 7 days to the experimental animals followed by LPS (10 mg/kg) injection interaperitonially. After LPS induction, blood was collected and the haematological parameters were analysed followed by isolation of heart tissue for biochemical molecular and histopathological analysis Neophytadiene significantly inhibited the NO production and inflammatory cytokines TNF-α, IL-6 and IL-10 both in in vitro and in vivo conditions. Further, the expression of TNF-α, IL1ß, NF-κB, iNOS, PI3k/Akt and MAPK in the heart tissue was modulated by Neophytadiene significantly confirming the anti-inflammatory potential. Thus, the effect of Neophytadiene on LPS-induced cardiac injury can be attributed to its anti-inflammatory antioxidant and cardioprotective properties.

Protective effect of rutin isolated from Spermococe hispida against cobalt chloride-induced hypoxic injury in H9c2 cells by inhibiting oxidative stress and inducing apoptosis.

BACKGROUND: Cardiovascular 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/PURPOSE: Reportedly, 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-METHODS: Phytochemical methods and advanced analytical methods were employed to isolate natural products from Spermococe hispida their effects in cardiomyocyets. RESULTS: We 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. CONCLUSION: The 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.

[6]-Gingerol-induced cell cycle arrest, reactive oxygen species generation, and disruption of mitochondrial membrane potential are associated with apoptosis in human gastric cancer (AGS) cells.

Ginger (Zingiber officinale Roscoe), a monocotyledonous herb, is widely used as an herbal medicine owing to the phytoconstituents it possesses. In the current study, the quantity of [6]-gingerol, the major phenolic ketone, in the fresh ginger and dried ginger rhizome was found to be 6.11 µg/mg and 0.407 µg/mg. Furthermore, [6]-gingerol was assessed for its antiapoptotic effects in human gastric adenocarcinoma (AGS) cells evidenced by acridine orange/ethidium bromide staining technique and Annexin-V assay. An increase in reactive oxygen species (ROS) generation led to a decrease in mitochondrial membrane potential (MMP) and subsequent induction of apoptosis. Results disclose that perturbations in MMP are associated with deregulation of Bax/Bcl-2 ratio at protein level, which leads to upregulation of cytochrome-c triggering the caspase cascade. These enduringly suggest that [6]-gingerol can be effectively used for targeting the mitochondrial energy metabolism to manage gastric cancer cells.

Inhibition of quorum sensing-controlled biofilm formation in Pseudomonas aeruginosa by quorum-sensing inhibitors.

Antimicrobial therapy against extensively drug-resistant (XDR) P. aeruginosa biofilms is less efficient compared to the treatment of equal bacterial counts of free-floating planktonic cells, which has become a serious threat in hospital environment. P. aeruginosa regulate their cooperative activities and physiological processes through a cell to cell chemical communication process called Quorum sensing (QS). This attracted our interest to synthesize, and to chemically characterize two anti-QS compounds, N-(4-{4-fluoroanilno} butanoyl) -l-homoserine lactone (FABHL) and N-(4-{4-chlororoanilno} butanoyl) -l-homoserine lactone (CABHL) to inhibit biofilm formation via disabling the QS circuits. Structural and morphological properties of these compounds were characterized by 1H Nuclear Magnetic Resonance (NMR), 13C NMR and High-resolution mass spectrometry (HRMS). Two biofilm forming XDR P. aeruginosa isolates were included in this study. Anti-biofilm property of FABHL or CABHL was confirmed by biofilm formation assay and it was shown to occur without affecting the bacterial growth. Anti-QS property of FABHL or CABHL was determined by evaluating the expression levels of QS genes (lasR and rhlR) by quantitative real time PCR (qRT-PCR). Although, FABHL and CABHL downregulates the expression levels of QS genes, lasR expression was significantly reduced. Molecular modeling studies revealed that the binding energy of FABHL and CABHL with LasR protein was -4.27 and -4.51, respectively. Hence, the synthesized compounds have the potential to serve as a potent anti-biofilm agent via disabling the QS systems. Lethality of FABHL and CABHL against PBMCs was assessed by 3-(4, 5- dimethylthiazol-2-yl)-2, 5-diphynyl tetrazolium bromide (MTT) assay. Cell viability was observed for both the compounds.

Attenuation of inflammation by marine algae Turbinaria ornata in cotton pellet induced granuloma mediated by fucoidan like sulphated polysaccharide.

Turbinaria ornata, a commonly found marine brown algae along the Gulf of Mannar, Southeast coast of India was evaluated for its anti-inflammatory potential and the bioactive compound present in it was characterized. Cotton pellet induced granuloma model in rats was used to assess the anti-inflammatory potential of the aqueous extract of Turbinaria ornata (ATO) (30, 100 and 300mg/kg, p.o) which was compared with dexamethasone (0.1mg/kg, p.o) a standard anti-inflammatory agent. Granuloma weight, haematological parameters and plasma markers (LDH, GPT, and CRP) were estimated. Further, the levels of oxidative stress markers (SOD, GPx, GSH, LPO, and Nitrite) and inflammatory markers (Cathepsin D and MPO) in the hepatic tissue were measured. ATO decreased the granuloma weight dose dependently. ATO significantly reversed the levels of biochemical and inflammatory markers in comparison to the vehicle treated rats. The active constituent, fucoidan (sulphated polysaccharide) from the aqueous extract was fractionated and characterized using GCMS. The sulphated polysaccharide (TSP) from ATO confirms the presence of sulphates and sugars. The present findings suggest ATO to be a potent inhibitor of both proliferative and exudative phases of inflammation possibly mediated by the sulphated polysaccharides which might inhibit the action of COX-2 enzyme analogous to dexamethasone.

PPAR's and Diosgenin a chemico biological insight in NIDDM.

Diosgenin a steroidal saponin found widely in nature is reported to contain several biological activities in recent years. The present work elaborates the modulation of the lipid and antioxidant profile by Diosgenin in diabetic condition. Type 2 diabetes was induced in experimental animals by feeding high fat diet (HFD) for 8 weeks followed by streptozotocin (STZ) injection (sub-diabetogenic dose; 35 mg/kg body weight). Diosgenin administered orally at two doses (40 and 80 mg/kg body weight) for 14 days reduced hyperglycemia, hypercholesterolemia and hypertriglyceridemia (p<0.001). Oxidative stress a crucial marker of diabetes and obesity associated complications was analyzed and noteworthy changes were observed. Improved levels of the antioxidant enzymes SOD and GPx and a minimized level of lipid peroxidation were also observed in Diosgenin treated rats. Further, analyzing the lipid accumulation by Oil Red O staining in 3T3-L1 preadipocytes confirmed its adipogenic activity which was influenced by PPAR γ and PPAR α. This was also substantiated through docking studies of Diosgenin with the PPARs. Altogether, Diosgenin a phytochemical of natural origin is found to mitigate diabetes induced oxidative stress and dyslipidemia which is crucial in cardio-metabolic risks by modulating the PPARs.

Sub-acute toxicity profile of a modified resveratrol supplement.

Longevinex, a nutraceutical formulation containing Resveratrol as the main component along with other polyphenolics exhibits diverse health benefits but systemic safety studies are lacking. Hence, to test the safety of Longevinex use for therapeutic purposes, 50 Sprague Dawley rats were randomly divided into five groups (n=10; 5M, 5F) wherein group I as vehicle treated control, group II and group III received 50 mg and 100 mg of plain Resveratrol respectively and group IV and group V received 50 mg and 100 mg of Longevinex respectively for a period of 28 days. All toxicological parameters were analyzed as per OECD-407 guidelines. Results showed treatment with Resveratrol and Longevinex did not result in any mortality of rats neither did they exhibit any clinical signs of toxicity. Hematological and biochemical analysis of serum enzymes and metabolites were not significantly altered between Longevinex and control rats. Likewise, histopathological analysis for various organs did not reveal significant changes in the vital organs of the treated rats. The study revealed that there were no significant treatment related adverse effects in rats exposed to Longevinex for 28 days and considered safe at the given dose where compared to plain Resveratrol.