Pomegranate (Punica granatum) purified polyphenol extract inhibits influenza virus and has a synergistic effect with oseltamivir

The pomegranate (Punica granatum L.) fruit has a long history of human consumption and possesses notable antioxidant and cardiovascular properties. This work evaluated the feasibility to provide a new functional beverage based on a dealcoholized red wine matrix supplemented by a pomegranate extract. The potential bioactive compounds in the pomegranate extract, punicalagin A and B and ellagic acid, were analyzed during the downstream process in order to evaluate the functional dose in the final beverage. The addition of pomegranate extract to the dealcoholized red wine resulted in a product with more intense yeast odor, acidity, yeast flavor, and astringency and with a less intense berry flavor. Consumer acceptance of the product was also investigated and the results revealed the existence of a niche of consumers willing to consume dealcoholized wine enriched with pomegranate extract. After tasting, 50% and 40% of those consumers initially interested by this product concept declared to be interested to purchase the control sample and the functional beverage, respectively. The daily consumption of two servings of 250 mL of this new pomegranate-enriched dealcoholized wine provides 82 mg of total ellagitannins, corresponding to the sum of punicalagin A and B and ellagic acid.

Despite rising interest in understanding intestinal bacterial survival in situ, relatively little attention has been devoted to deciphering the interaction between bacteria and functional food ingredients. Here, we examined the interplay between diverse beneficial Lactobacillaceae species and a pomegranate (POM) extract and determined the impact of this functional ingredient on bacterial growth, cell survival, transcription and target metabolite genesis. Three commercially available probiotic strains (Lactobacillus acidophilus NCFM, Lacticaseibacillus rhamnosus GG and Lactiplantibacillus plantarum Lp-115) were used in growth assays and flow cytometry analysis, indicating differential responses to the presence of POM extract across the three strains. The inclusion of POM extract in the growth medium had the greatest impact on L. acidophilus cell counts. LIVE/DEAD staining determined significantly fewer dead cells when L. acidophilus was grown with POM extract compared to the control with no POM (1.23% versus 7.23%). Whole-transcriptome analysis following exposure to POM extract showed markedly different global transcriptome responses, with 15.88% of the L. acidophilus transcriptome, 19.32% of the L. rhamnosus transcriptome and only 2.37% of the L. plantarum transcriptome differentially expressed. We also noted strain-dependent metabolite concentrations in the medium with POM extract compared to the control medium for punicalagin, ellagic acid and gallic acid. Overall, the results show that POM extract triggers species-specific responses by probiotic strains and substantiates the rising interest in using POM as a prebiotic compound.

Background and Aim:It has long been known that the spermatogenic tissue is very sensitive to temperatures higher than its physiologic temperature and causing cessation of activity and resulting in sterility. This study investigated the effect of a standardized 40% ellagic acid extract of pomegranate on the histopathology, diameter, and epithelial thickness of seminiferous tubules in albino rats exposed to heat.Materials and Methods:Twenty-five male albino Wistar rats were randomized at 7-8 months of age to five treatment groups. Group C was not treated; Group T0 was treated with 0.5% of Na carboxymethyl cellulose (CMC) 2 ml/day and exposed to heat. T1, T2, and T3 were treated with 75, 150, and 300 mg/kg/day of a standardized 40% ellagic acid extract of pomegranate (Punica granatum L.), respectively. The animals were orally administered Na CMC or pomegranate extract and were exposed to sunlight for 15 min at 40°C-42°C for 14 days. The animals were sacrificed on day 15 and the testes were removed for histological evaluation and measurement of seminiferous tubule diameter and epithelium thickness.Results:The diameter of seminiferous tubules from rats exposed to heat and treated with 300 mg/kg/day pomegranate extract was larger and the epithelia thicker than those in the other groups (p<0.05). The protective effects of the standardized 40% ellagic acid extract may have been mediated by its antioxidant activity.Conclusion:Compared with controls, administration of 300 mg/kg/day of a standardized 40% ellagic acid extract of P. granatum L. for 14 days increased seminiferous tubule diameter and epithelium thickness in albino Wistar rats exposed to heat.

Pomegranate (Punica granatum) extract and its polyphenols reduce the formation of methylglyoxal-DNA adducts and protect human keratinocytes against methylglyoxal-induced oxidative stress

Eight cell lines were systematically compared for their permissivity to primary infection, replication, and spread of seven human influenza viruses. Cell lines were of human origin (Caco-2, A549, HEp-2, and NCI-H292), monkey (Vero, LLC-MK2), mink (Mv1 Lu), and canine (MDCK). The influenza viruses included seasonal types and subtypes and a pandemic virus. The MDCK, Caco-2, and Mv1 Lu cells were subsequently compared for their capacity to report neutralization titers at day one, three and six post-infection. A gradient of sensitivity to primary infection across the eight cell lines was observed. Relative to MDCK cells, Mv1 Lu reported higher titers and the remaining six cell lines reported lower titers. The replication and spread of the seven influenza viruses in the eight cell substrates was determined using hemagglutinin expression, cytopathic effect, and neuraminidase activity. Virus growth was generally concordant with primary infection, with a gradient in virus replication and spread. However, Mv1 Lu cells poorly supported virus growth, despite a higher sensitivity to primary infection. Comparison of MDCK, Caco-2, and Mv1 Lu in neutralization assays using defined animal antiserum confirmed MDCK cells as the preferred cell substrate for influenza virus testing. The results observed for neutralization at one day post-infection showed MDCK cells were similar (<1 log2 lower) or superior (>1 log2 higher) for all seven viruses. Relative to Caco-2 and Mv1 Lu cells, MDCK generally reported the highest titers at three and six days post-infection for the type A viruses and lower titers for the type B viruses and the pandemic H9N2 virus. The reduction in B virus titer was attributed to the complete growth of type B viruses in MDCK cells before day three post-infection, resulting in the systematic underestimation of neutralization titers. This phenomenon was also observed with Caco-2 cells.

Dietary supplementation of an ellagic acid-enriched pomegranate extract attenuates chronic colonic inflammation in rats

Aurantiamide acetate from baphicacanthus cusia root exhibits anti-inflammatory and anti-viral effects via inhibition of the NF-κB signaling pathway in Influenza A virus-infected cells

BackgroundThe hemagglutinin (HA) of influenza viruses is a possible target for antiviral drugs because of its key roles in the initiation of infection. Although it was found that a natural compound, Stachyflin, inhibited the growth of H1 and H2 but not H3 influenza viruses in MDCK cells, inhibitory activity of the compound has not been assessed against H4-H16 influenza viruses and the precise mechanism of inhibition has not been clarified.MethodsInhibitory activity of Stachyflin against H4-H16 influenza viruses, as well as H1-H3 viruses was examined in MDCK cells. To identify factors responsible for the susceptibility of the viruses to this compound, Stachyflin-resistant viruses were selected in MDCK cells and used for computer docking simulation.ResultsIt was found that in addition to antiviral activity of Stachyflin against influenza viruses of H1 and H2 subtypes, it inhibited replication of viruses of H5 and H6 subtypes, as well as A(H1N1)pdm09 virus in MDCK cells. Stachyflin also inhibited the virus growth in the lungs of mice infected with A/WSN/1933 (H1N1) and A/chicken/Ibaraki/1/2005 (H5N2). Substitution of amino acid residues was found on the HA2 subunit of Stachyflin-resistant viruses. Docking simulation indicated that D37, K51, T107, and K121 are responsible for construction of the cavity for the binding of the compound. In addition, 3-dimensional structure of the cavity of the HA of Stachyflin-susceptible virus strains was different from that of insusceptible virus strains.ConclusionAntiviral activity of Stachyflin was found against A(H1N1)pdm09, H5, and H6 viruses, and identified a potential binding pocket for Stachyflin on the HA. The present results should provide us with useful information for the development of HA inhibitors with more effective and broader spectrum.

Madin-Darby canine kidney (MDCK) cells are one of the main cell lines used for influenza vaccine production due to their high virus yield and low mutation resistance. Due to their high tumorigenicity, the safety of vaccines produced from these cells is controversial. TGM2 is a multifunctional protein that plays an important role in the adhesion and migration of cells and is associated with tumor formation. We found that the expression level of TGM2 was significantly up-regulated in low tumorigenic MDCK cells. We first analyzed TGM2-overexpressed and knockout MDCK cells in vitro. Scratch-wound assay and Transwell chamber experiments showed that TGM2 overexpression significantly inhibited the migration and invasion of MDCK cells and significantly reduced their proliferation. TGM2 knockout significantly enhanced cell migration, invasion, and proliferation. The tumorigenesis results in nude mice were consistent with those in vitro. TGM2 knockout significantly enhanced the tumorigenesis rate of MDCK cells in nude mice. We also investigated the effects of TGM2 gene expression on the replication of the H1N1 influenza A virus in MDCK cells. The results showed that TGM2 induced the negative regulation of H1N1 replication. These findings contribute to a comprehensive understanding of the tumor regulation mechanism and biological functions of TGM2.

BackgroundS-033447, an active form of orally available prodrug S-033188, is a novel small molecule inhibitor of cap-dependent endonuclease that is essential for influenza virus transcription and replication. In this study, we evaluated the inhibitory effect of S-033188 in combination with neuraminidase inhibitors on the replication of influenza A/H1N1 virus in cultured cells.MethodsThe inhibitory effects of S-033447 in combination with NA inhibitors on the cytopathic effect of A/PR/8/34 strain in Madin–Darby canine kidney cells cultured for 2 days were tested and EC50 were determined. The combination index (CI), which were obtained when S-033188 and NA inhibitor were added at the closest ratio of each EC50 value, were used for the evaluation of these combinational effects (Table 1). CI values were calculated by the Chou and Talalay method, in which combinational effect were determined according to the criteria as follows: synergistic if CI ≤ 0.8, additive if 0.8 < CI < 1.2, and antagonistic if CI ≥ 1.2.CI = (DA/A + B)/DA + (DB/A + B)/DB + (DA/A + B × DB/A + B)/(DA × DB)DA: the EC50 of S-033447DB: the EC50 of NA inhibitorDA/A + B: the concentration of S-033447 giving 50% inhibition in combination with NA inhibitor at the closest ratio of each EC50 valueDB/A + B: the concentration of NA inhibitor giving 50% inhibition in combination with S-033447 at the closest ratio of each EC50 valueResultsAll CI values were lower than 0.8, under the condition that both S-033447 and NA inhibitor (oseltamivir acid, zanamivir hydrate, laninamivir, or peramivir trihydrate) were added at the closest ratio of each EC50 value (Table 1).ConclusionS-033447 in combination with oseltamivir acid, zanamivir hydrate, laninamivir, or peramivir trihydrate synergistically inhibited the replication of influenza A/H1N1 virus in MDCK cells.Table 1. Combination effect of S-033447 and NA inhibitor in MDCK cells infected with A/PR/8/34 strainSubstance ASubstance BDA (nmol/L)DB (nmol/L)DA/A+B (nmol/L)DB/A+B (nmol/L)CICombination effectS-033447 oseltamivir acid4.513171.971.17586.940.49synergisticS-033447zanamivir hydrate4.491565.381.22305.990.52synergisticS-033447laninamivir4.52212.741.1256.020.58synergisticS-033447 peramivir trihydrate4.41213.771.1356.660.59synergisticDisclosuresAll authors: No reported disclosures.

Flaviviruses include the most prevalent and medically challenging viruses. Persistent infection with flaviviruses of epithelial cells and hepatocytes that do not undergo cell death is common. Here, we report that, in epithelial cells, up-regulation of autophagy following flavivirus infection markedly enhances virus replication and that one flavivirus gene, NS4A, uniquely determines the up-regulation of autophagy. Dengue-2 and Modoc (a murine flavivirus) kill primary murine macrophages but protect epithelial cells and fibroblasts against death provoked by several insults. The flavivirus-induced protection derives from the up-regulation of autophagy, as up-regulation of autophagy by starvation or inactivation of mammalian target of rapamycin also protects the cells against insult, whereas inhibition of autophagy via inactivation of PI3K nullifies the protection conferred by flavivirus. Inhibition of autophagy also limits replication of both Dengue-2 and Modoc virus in epithelial cells. Expression of flavivirus NS4A is sufficient to induce PI3K-dependent autophagy and to protect cells against death; expression of other viral genes, including NS2A and NS4B, fails to protect cells against several stressors. Flavivirus NS4A protein induces autophagy in epithelial cells and thus protects them from death during infection. As autophagy is vital to flavivirus replication in these cells, NS4A is therefore also identified as a critical determinant of flavivirus replication.

Anti-influenza virus phytochemicals from Radix Paeoniae Alba and characterization of their neuraminidase inhibitory activities

Change in receptor-binding specificity of recent human influenza A viruses (H3N2) affects recognition of the receptor on MDCK cells

Towards a yeast cell cycle Sys-bio model. Regulation of family 3 ubiquitin-conjugating enzymes by phosphorylation: a molecular dynamics investigation

Background: A vital and significant goal in treating a tooth with an apical infection is getting rid of the germs in the pulp space. Recurrent root canal infections are most frequently linked to Enterococcus faecalis and can happen even after endodontic therapy. E. faecalis can resist routine endodontic disinfectants and can also survive the nutrient-deprived conditions in the root filled tooth. So, an attempt has been made to eliminate the microorganisms using the herbal extracts which have antimicrobial properties and comparing it with the well accepted and excellent antimicrobial agent 2% chlorhexidine. Aim: To compare Syzygium aromaticum (Clove) and Punica granatum (Pomegranate) extracts with 2% chlorhexidine in dentinal tubule disinfection with Real-time polymerase chain reaction which was used to detect E. faecalis. Methods: Thirty-six extracted premolar teeth were selected, access cavity was prepared and cleaning and shaping was done. With the help of a rotating diamond disc bur, the middle part of the root was sliced. E. faecalis was applied to the tooth specimens and left on them for 21 days. Group 1 specimens had pomegranate extract, Group 2 contained clove extract, and Group 3 contained 2% CHX. After being watered by the corresponding groups, the specimens were incubated for 5 days. A Gates-Glidden drill was used to collect the dentinal shavings, which were then subjected to DNA isolation before being subjected to real-time PCR analysis. Statistical analysis used: The results were statistically analyzed using one way ANOVA and Post hoc Tukey's analysis. Results: Threshold cycle (Ct) values showed greater inhibition of bacterial load with pomegranate extracts followed by 2% chlorhexidine. Lesser reduction of bacterial load was found with clove extract. Conclusion: Pomegranate extract, an herbal extract with therapeutic potential which can be utilised as an efficient substitute for 2% CHX for treating E. fecalis. Keywords: Clove; Pomegranate; E. Fecalis; Real-time PCR.