Genomic DNA Isolation by Phenol/Chloroform Extracting Method from Sheep Blood Clot

[ Objective] The aim was to establish the method of extracting genomic DNA from sheep blood clot on the basis of the improvement of method for extracting genomic DNA from tissues. [Method]The genomic DNA with complete primary structure and high purity was obtained from the sheep blood clot after the steps of cutting the sheep blood clot with ophthalmic scissors, cell lysis with tissue DNA extracts and digested by proteinase K, extracting with phenol/chloroform and precipitating with ethanol were performed. [ Result] The concentration of the extracted DNA was 159.90 ±0.70 ng/μl and the ratio of the A260/A280 was 1.80 ＋0.01. The sheep microsatellite locus of BM203 was amplified by using the extracted DNA from the sheep blood clot as template of PCR, and the PCR result was perfect. [Conclusion]This method is simple and feasible, the quantity and quality of the extracted DNA can satisfy the demands for the subsequent researches. It is worth to extending and using for reference.

Phenolic extract of Morchella angusticeps peck inhibited the proliferation of HepG2 cells in vitro by inducing the signal transduction pathway of p38/MAPK

Morchella angusticeps Peck,one of the most popular edible mushrooms,has attracted great attention due to its delicious taste and healthy properties.However,both its biological effects and the possible mechanism of action have not yet been known.We investigated the anti-proliferative activity of the phenolic extract derived from Morchella angusticeps Peck(MPE)against HepG2 human hepatocellular carcinoma cells.Results showed that MPE at non-cytotoxicity doses significantly inhibited the proliferation of HepG2 cells in a dose-dependent manner with inhibitory rates ranging from 18 to 90%(P<0.01).The possible mechanism might be that MPE induced apoptosis through initiating the mitochondrial death pathway by regulating Bax,Bcl-2 and cleaved caspase-3.On the other hand,MPE might trigger cell cycle arrest at G0/G1/S phases by managing p21,Cyclin D1,cyclin-dependent kinases-4(CDK4)and proliferating cell nuclear antigen(PCNA).Additionally,MPE downregulated TRAF-2 and p-p53,while upregulated p-ASK1 and p-p38.Therefore,it could be inferred that MPE might induce the anti-proliferative function to HepG2 cells through the p38/MAPK signal transduction pathway.

Wild pink bayberry free phenolic extract induces mitochondria-dependent apoptosis and G0/G1 cell cycle arrest through p38/MAPK and PI3K/Akt pathway in MDA-MB-231 cancer cells

Polyphenol-rich foods have been shown to be good for cancer prevention as powerful antioxidants. In this study, the mechanisms of wild pink bayberry free phenolic extract(WPBFE)inhibiting the proliferation and inducing apoptotic of MDA-MB-231 breast cancer cells was examined. The main phenolic acids and flavonols in WPBFE were gallic acid((18.83 ± 0.44)μg/g FW)and myricetin((1.52 ± 0.05)μg/g FW), respectively. The maximum inhibition rate of WPBFE at non-cytotoxicity dose(below 80 mg/mL)was 81%. Western blotting analysis showed that WPBFE could cause the arrest of cell cycle in G0/G1 phase by down-regulating expression levels of PCNA, CDK4, cyclin D1 and up-regulating the expression level of p21. Meanwhile, WPBFE induced apoptosis through initiating the mitochondrial death pathway by up-regulating cleaved caspase-3 and enhancing the ratio of Bax/Bcl-2, with the maximum expression levels of 1.29 and 2.03 folds that of control group, respectively. Further study of the upstream protein, we found that WPBFE down-regulated TRAF2, while upregulated p-ASK1, p-p38 and p-p53. Furthermore, WPBFE could down-regulate the expression of p-PI3K and p-Akt. These observations indicated that WPBFE might play an anticancer role through regulating the p38 MAPK together with PI3K/Akt pathway.

Phenolic compounds from Chaenomeles speciosa alleviate inflammation in lipopolysaccharide-treated RAW264.7 macrophages via the NF-κB and MAPK pathways

Chaenomeles speciosa(Sweet)Nakai cultivated widely in temperate regions possesses anti-inflammatory properties,however,the underlying molecular mechanisms remain not fully understood.In this study,a purified phenolic extract of C.speciosa rich in chlorogenic acid,procyanidin B1 and catechin(determined by HPLC-Q-TOF-MS/MS)exhibited dose-dependent anti-inflammatory effects on lipopolysaccharide(LPS)-treated RAW264.7 macrophages.The extract at 30μg/mL was most potent and enabled most cells in normal morphology under LPS stimulation without causing cytotoxicity.The extract suppressed the levels of nitric oxide(NO),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and interleukin-1β(IL-1β),and the mRNA and protein expressions of nitric oxide synthase(iNOS)and cyclooxygenase-2(COX-2).The mechanisms underlying such anti-inflammatory actions included the regulation of phosphorylation of related proteins to monitor the expressions of inflammatory mediators and genes in the nuclear factor kappa-B(NF-κB)and mitogen-activated protein kinase(MAPK)signaling pathways.Therefore,the phenolic extract from C.speciosa is a desirable anti-inflammatory agent for inflammatory conditions to meet the rising demand for natural and cost-effective therapeutics.

Discovery of possible hepatoprotective components from lychee pulp phenolic extract by online knockout methods

Lychee pulp phenolic extract (LPPE) possessed protective effects against nonalcoholic fatty liver disease (NAFLD), while the specific components concerned were unclear. In this study, twelve major components in LPPE were identified by LC-MS and prepared by on line knockout methods. Two antioxidant methods companied with hepatocyte steatosis assay were used to evaluate the contribution of twelve components to the hepatoprotective effects of LPPE. The identified components include four procyanidins and eight flavonoids, they all play active roles in the oxygen radical absorbance capacity of LPPE, while only B-type procyanidin trimer and quercetin 3-O -rutinoside-7-O -rhamnoside exert positive effects on cellular antioxidant activity of LPPE. Six components including procyanidin B2, epicatechin, two quercetin glycosides, and two procyanidin trimers are considered as contributors to lipid accumulation inhibitory effects of LPPE with B-type procyanidin trimer being identified as the main contributor. All the twelve components were recognized as hepatoprotective components against NAFLD in LPPE.

Cinnamon free phenolic extract regulates glucose absorption in intestinal cells by inhibiting glucose transporters

Cinnamon is a traditional herbal medicine that is a valuable source of bioactive phenolic compounds.In this study,we determined the effects of cinnamon free phenolic extract(CFPE)on glucose transport in Caco-2 cells and its possible mechanisms of action.First,the contents of phenolic compounds and flavonoids in bark were compared among four cinnamon cultivars.The‘Taiwan’cultivar had the highest contents of these compounds,and the CFPE was prepared from it.A Caco-2 monolayer was established,and the effect of CFPE on glucose transport across the monolayer was determined.The results showed that CFPE(at 150-300μg/mL)suppressed glucose transport across the monolayer in a dose-dependent manner.Procyanidin C1,procyanidin B2,procyanidin B1,procyanidin B3,procyanidin A2,and(􀀀)-epicatechin derivatives in CFPE may have contributed to its suppressive effect.Treatment with CFPE decreased the transcript levels of SGK1 encoding serum and glucocorticoid-inducible kinase and PLC/PKC encoding phospholipase C/protein kinase C,which are involved in the signaling pathway associated with glucose transport;and the transcript levels of SGLT1 encoding sodium/glucose cotransporter 1 and GLUT2 encoding glucose transporter 2,the key glucose transporters in this pathway.These findings suggest that CFPE inhibits glucose transport via its effects on the signaling pathway and glucose transporters involved in glucose absorption in intestinal cells.Thus,CFPE has potential applications in preventing postprandial hyperglycemia.

Antioxydant Activity of Some Algerian Olive Oil Samples in the Eastern and Center of Algeria

This study aimed to evaluate the antioxidant activity of five Algerian olive oil samples from Chlef, Jijel, Tissemsilt, Bejaia and Tizi Ouzou regions. Thus the following results were found physicochemical caracterisation revealed that the samples are conform to standards prescribed by IOC （2013） as they are type-oleic and linoleic varies between virgin and extra virgin Chlef, Djijel and Bejaia oils are richer in carotenoids than those ofTizi Ouzou and Tissemsilt. Chlefand Tissemsilt oils are richer in phenolic compounds than other oils and have revealed very important antioxidant activity and the largest inhibitory power of peroxidation.

Pomegranate （Punica granatum L.） Peel Extract as a Source of Natural Antioxidant

An increase in the demand for natural additives has shifted the attention from synthetic to natural antioxidants. Antioxidative components from pomegranate peel were extracted by using different solvents like methanol, ethanol, acetone, chloroform and ethyl acetate. All the extracts were tested for Total phenolic content （TPC） and the methanol which proved to be best in extracting the phenolic compounds was used to assess the antioxidant property by using different methods like Diphenyl piridyl picryl hydrazyl （DPPH）, Relative reducing power （RRP）, Beta carotene bleaching and Ferric thiocyanate （FTC）. The antioxidant activity （AA） of pomegranate peel extract （PPE） was compared with Butylated hydroxyl toluene （BHT） and Butylated hydroxyl anisole （BHA）. The methanolic extract was examined by Liquid Chromatography and Mass Spectroscopy （LCMS） and the spectra showed the presence of different phenolic compounds. The extracts were tested for their efficiency in preventing rancidity of Refined Bleached Deodorized Soybean oil （RBDSBO） in comparison with BHT and control at 200 ppm concentration. The PPE proved to be best in preventing rancidity. Owing to the present results, the studies can further be extended to exploit the PPE for application into different food products.

Free Radical-Scavenging Properties and Antioxidant Activity of Fractions from Cranberry Products

Lipid peroxidation inhibition capacity and antiradical activity were evaluated in HPLC fractions of different polarity obtained from two cranberry juices and three extracts isolated from frozen cranberries and pomace containing antho-cyanins, water-soluble and apolar phenolic compounds, respectively. Compounds with close polarities were collected to obtain between three and four fractions from each juice or extract. The cranberry phenols are good free radi-cal-scavengers, but they were less efficient at inhibiting the lipid peroxidation. Of all the samples tested, the intermediate polarity fraction of extract rich in apolar phenolic compounds of fruit presented the highest antiradical activity while the most hydrophobic fractions of the anthocyanin-rich extract from fruit and pomace appeared to be the most efficient at inhibiting the lipid peroxidation. The antioxidant or pro-oxidant activity of fractions increased with the con-centration. The phenol polarity and the technological process to manufacture cranberry juice can influence the antioxidant and antiradical activities of fractions.

Potential Application of a Wine Extract in Skin Care: How to Benefit from the Antibacterial, Antioxidant and Elastase Inhibiting Properties

Since plant polyphenols have many beneficial properties on health, the aim of this study was to evaluate the potential use of a phenolic wine extract, a by-product of wine production, for skin care on HaCaT cells. In these studies, a significant reduction of reactive oxygen species formation in HaCaT cells and severe elastase inhibition was observed. In contrast, the wine extract caused a major increase in lipase activity. The extract showed no influence on cell proliferation, but an immunomodulatory effect on the release of the interleukins IL-6 and IL-8 was found. The phenolic wine extract demonstrated a strong activity against gram-positive and gram-negative pathogens, yeasts, and fungi. Overall, our results show that the investigated phenolic wine extract is a promising ingredient for anti-aging skin care, could contribute to the improvement of skin appearance and health, and may positively affect cellulite.

Fermented Psidium guajava leaves regulate the gut microbiota and improve metabolic alterations in diabetic mice

The phenolic extract from fermented Psidium guajava leaves(PFGL)helps to improve diabetes;however,its hypoglycemic and hypolipidemic mechanisms remain unclear.In this study,gut microbiota modulation and glucose-lipid metabolism was investigated.The results showed that dietary supplement of PFGL enriched in quercetin glucosides significantly improved serum glucose-lipid profiles as well as insulin resistance in diabetic mice.The gut microbiota of diabetic mice was modulated by PFGL(200 mg/kg),causing an abundance of Bacteroidetes,Prevotellaceae,and Alistipes and a reduced the presence of Firmicutes and Streptococcus.Increased levels of short-chain fatty acids(SCFA)activated G protein-coupled receptor signaling that induced peptide YY release to promote insulin secretion.Diabetes-induced serum metabolite disorders that are involved in glycerophospholipid metabolism and tryptophan metabolism were restored by PFGL.The protein expression of metabolic enzymes including phospho-adenosine 5’-monophosphate-activated protein kinase,fatty acid synthase,acyl-coenzyme A oxidase,and diacylglycerol aeyltransferase 2 were also regulated to promote glucoselipid metabolism.In particular,gut microbiota enriched with Alistipes genera and SCFA metabolic regulation were closely correlated with diabetes-related indicators.These findings provide a theoretical support for the development and application of PFGL for the treatment of diabetes.