Compositional analysis of the glycosaminoglycan family in velvet antlers of Sika deer (Cervus nippon) at different growing stages.

Glycosaminoglycans (GAG) from the velvet antlers of Sika deer (Cervus nippon) at the different growing stages (Fukurozuno, Anshi, and Santajo) of bred and wild deer were isolated and their concentrations and sulfation patterns were analyzed. GAG were digested with chondroitinase ABC, ACI, heparinase-I and -III, and keratanase-II into the corresponding repeating disaccharides of chondroitin sulfate (CS), dermatan sulfate (DS), hyaluronan, heparan sulfate (HS), and keratan sulfate. Cartilaginous tissues contained CS-DS at high concentrations with an almost equal ratio of 4- and 6-sulfates, while 4-sulfate-type CS-DS predominantly occupied ossified tissues, but at low concentrations. High O- and N-sulfation degrees of HS correspond to high ossification. Dynamic quantitative changes in CS-DS and compositional changes in CS-DS and HS were closely associated with the mineralization of deer antlers.

Fermented Asterina pectinifera with Cordyceps militaris Mycelia Induced Apoptosis in B16F10 Melanoma Cells.

This prime objective of this study was to explore the anti-cancer activity of fermented Asterina pectinifera with Cordyceps militaris mycelia (FACM) in B16F10 murine melanoma cells. The effect of FACM on cell viability was assessed using MTT assay. Furthermore, the effect of FACM was compared with unfermented A. pectinifera on cell viability. The results demonstrated that the fermented FACM extract has a higher inhibitory activity on the proliferation of B16F10 murine melanoma cells than unfermented A. pectinifera. In addition, FACM also promoted the expression of pro-apoptotic protein Bax leading to stimulate apoptosis in B16F10 cells. Therefore the present study demonstrates that the FACM might be a potential effective anti-cancer agent, as a result of its stronger anti-proliferative effect and apoptosis inducing effect than A. pectinifera or C. militaris on melanoma cells.

Antioxidant Activity of Extract from the Cephalothorax of Fenneropenaeus chinensis.

We investigated the antioxidant activity of taurine rich water extract from the cephalothorax of Fenneropenaeus chinensis (FCC). The antioxidant potency of water extract from FCC was assessed using various assay methods, such as DPPH (1,1-diphenyl-2-picrylhydrazyl), alkyl radical scavenging activity, ABTS (2,2'-azinobis (3-ethylbenzothiazoline 6-sulfonic acid ammonium salt)) radical scavenging activity and Ferric reducing antioxidant power (FRAP) assay. The DPPH and alkyl radical scavenging activities of FCC were dose-dependently increased. The lipid peroxidation was estimated using ferric thiocyanate (FTC) assay and thiobarbituric acid (TBA) methods. However, a higher lipid peroxidation activity was observed in TBA method than FTC method. The results of the present study suggested that the FCC extract potentially scavenged the free radical and reduced oxidative stress. Therefore, the present study is concluded that the FCC extract could be a potential source of antioxidant activity.

Antioxidant Effect of Taurine-Rich Paroctopus dofleini Extracts Through Inhibiting ROS Production Against LPS-Induced Oxidative Stress In Vitro and In Vivo Model.

Taurine is an essential amino acid to improve the function of cardiovascular, skeletal muscle, retina, and central nervous system. It also plays a role as an antioxidant agent against reactive oxygen species (ROS) generated by various substances. The aim of the current study was to examine the antioxidant capacity of water extracts of Paroctopus dofleini. Radical scavenging activity of P. dofleini extracts was performed using an ESR spectrophotometer. Protective effects of P. dofleini extracts against lipopolysaccharide (LPS)-induced oxidative stress in RAW264.7 cells were evaluated using flow cytometry. The P. dofleini extracts showed a potent antioxidant activity against LPS-induced oxidative stress on RAW264.7 cells. Furthermore, the in vivo antioxidant activity of P. dofleini extract on LPS-induced oxidative stress was assessed using zebrafish embryos. P. dofleini successfully scavenged the LPS-induced intracellular ROS and prevented lipid peroxidation in zebrafish embryos. The results obtained in this study clearly demonstrate that the P. dofleini significantly scavenge the ROS and prevent lipid peroxidation in both in vitro and in vivo models.

Taurine Attenuates Doxorubicin-Induced Toxicity on B16F10 Cells.

This study aimed to investigate the effect of doxorubicin co-treatment with taurine on B16F10 melanoma cells. Frequently, Doxorubicin is used in the treatments of many different kinds of cancers, some of which are soft tissue sarcomas, hematological malignancies and carcinomas. However, the clinical application of doxorubicin is compromised by its severe adverse effects, including cardiotoxicity. In the present study, the efficacy of doxorubicin co-treatment with taurine was investigated. B16F10 cell viability was evaluated using MTT assays, trypan blue dye exclusion assays, and fluorescent staining technique. Apoptotic cells were detected by flow cytometry and the proteins associated with apoptosis and cellular differentiations were assessed by immunoblotting. Doxorubicin inhibited cell growth and induced cell death in B16F10 cells. Interestingly, doxorubicin co-treatment with taurine inhibited apoptosis in B16F10 cells. These results indicate that doxorubicin co-treatment with taurine attenuates doxorubicin-induced cytotoxicity and reduces ROS production in B16F10 cells.

Neuroprotective Effect of Taurine-Rich Cuttlefish (Sepia officinalis) Extract Against Hydrogen Peroxide-Induced Oxidative Stress in SH-SY5Y Cells.

Oxidative stress mediates the cell damage in several neurodegenerative diseases, some of which are Alzheimer's disease (AD), multiple sclerosis and Parkinson's disease (PD). In this study, we investigated whether the taurine-rich cuttlefish extract could exert a protective effect on damaged human neuroblastoma SH-SY5Y cells induced by hydrogen peroxide (H2O2). Our results revealed that pre-treatment with cuttlefish extract effectively increased the cell viability by protecting the cells from intracellular reactive oxygen species (ROS) induced by H2O2 exposure. Furthermore, apoptosis related proteins Bcl-2 and Bax were investigated by western-blot analysis and results indicated that cuttlefish extract promoted the expression of anti-apoptotic Bcl-2 protein while inhibiting the expression of pro-apoptotic Bax protein. Therefore, cuttlefish extract containing the ability of scavenging excessive ROS, the capacity of anti-oxidative stress, could be employed in neurodegenerative disease prevention. In conclusion, the results suggest that cuttlefish extract could be used as a potential candidate for preventing several human neurodegenerative and other disorders caused by oxidative stress.

Biosynthesis of Oligomeric Anthocyanins from Grape Skin Extracts.

We synthesized oligomeric anthocyanins from grape skin-derived monomeric anthocyanins such as anthocyanidin and proanthocyanidin by a fermentation technique using Aspergillus niger, crude enzymes and glucosidase. The biosyntheses of the oligomeric anthocyanins carried out by the conventional method using Aspergillus niger and crude enzymes were confirmed by ESI-MS. The molecular weight of the synthesized anthocyanin oligomers was determined using MALDI-MS. The yield of anthocyanin oligomers using crude enzymes was higher than that of the synthesis using Aspergillus fermentation. Several studies have been demonstrated that oligomeric anthocyanins have higher antioxidant activity than monomeric anthocyanins. Fermentation-based synthesis of oligomeric anthocyanins is an alternative way of producing useful anthocyanins that could support the food industry.

Trapa japonica Pericarp Extract Reduces LPS-Induced Inflammation in Macrophages and Acute Lung Injury in Mice.

In this study, we found that chloroform fraction (CF) from TJP ethanolic extract inhibited lipopolysaccharide (LPS)-induced production of nitric oxide (NO) and intracellular ROS in RAW264.7 cells. In addition, expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) genes was reduced, as evidenced by western blot. Our results indicate that CF exerts anti-inflammatory effects by down-regulating expression of iNOS and COX-2 genes through inhibition of MAPK (ERK, JNK and p38) and NF-κB signaling. Similarly we also evaluated the effects of CF on LPS-induced acute lung injury. Male Balb/c mice were pretreated with dexamethasone or CF 1 h before intranasal instillation of LPS. Eight hours after LPS administration, the inflammatory cells in the bronchoalveolar lavage fluid (BALF) were determined. The results indicated that CF inhibited LPS-induced TNF-α and IL-6 production in a dose dependent manner. It was also observed that CF attenuated LPS-induced lung histopathologic changes. In conclusion, these data demonstrate that the protective effect of CF on LPS-induced acute lung injury (ALI) in mice might relate to the suppression of excessive inflammatory responses in lung tissue. Thus, it can be suggested that CF might be a potential therapeutic agent for ALI.

Antioxidant and Anti-Inflammatory Effects of Chaenomeles sinensis Leaf Extracts on LPS-Stimulated RAW 264.7 Cells.

The fruit of Chaenomeles sinensis has been traditionally used in ethnomedicine for the treatment of various human ailments, including pneumonia, bronchitis, and so on, but the pharmacological applications of the leaf part of the plant have not been studied. In this study, we evaluated the various radical scavenging activities and anti-inflammatory effects of different Chaenomeles sinensis leaf (CSL) extracts. The water extract showed a higher antioxidant and radical scavenging activities. However the ethanolic extracts showed higher NO scavenging activity than water extract, therefore the ethanolic extract of CSL was examined for anti-inflammatory effects on lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. The 70% ethanol extract of CSL (CSLE) has higher anti-inflammatory activity and significantly inhibited the production of nitric oxide (NO), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α). In addition, CSLE suppressed LPS-stimulated inducible nitric oxide synthase (iNOS) and NO production, IL-1ß and phospho-STAT1 expression. In this study, we investigated the effect of CSLE on the production of inflammatory mediators through the inhibition of the TRIF-dependent pathways. Furthermore, we evaluated the role of CSLE on LPS-induced expression of pro-inflammatory cytokines, such as TNF-α, IL-1ß and IL-6. Our results suggest that CSLE attenuates the LPS-stimulated inflammatory responses in macrophages through regulating the key inflammatory mechanisms, providing scientific support for its traditional uses in treating various inflammatory diseases.

Effects of Physically Effective Neutral Detergent Fiber Content on Intake, Digestibility, and Chewing Activity in Fattening Heifer Fed Total Mixed Ration.

The objective of this study was to determine the effects of physically effective neutral detergent fiber (peNDF) content in total mixed ration (TMR) on dry matter intake, digestibility, and chewing activity in fattening Hanwoo (Bos taurus coreanae) heifers. The experiment was designed as a replicated 3×3 Latin square using 12 heifers. Fattening heifers were offered one of three diets [high (T1), medium (T2), and low (T3) peNDF] obtained by different mixing times (3, 10, and 25 min) for the same TMR feed. The peNDF content of TMR was determined by multiplying the proportion of dry matter retained by a 1.18 mm-screen in a Penn State Particle Separator by the dietary NDF content. The peNDF1.18 content was 30.36%, 29.20%, and 27.50% for the T1, T2, and T3 diets, respectively (p<0.05). Dry matter intake was not affected by peNDF content in TMR. Total weight gain in T1 group was significantly higher (p<0.05) than in T2 and T3 groups. However, weight gain did not differ between T2 and T3 groups. The feed conversion ratio decreased with an increase in the peNDF content (T1: 12.18, T2: 14.17, and T3: 14.01 g/g). An increase in the peNDF content of TMR was associated with a linear increase in the digestibility of dry matter, crude protein, crude fiber, neutral detergent fiber, and acid detergent fiber (p<0.05). Also, an increase in peNDF content of the TMR resulted in a linear increase in the number of chews in eating and ruminating (p<0.05), and consequently in the number of total chews (p<0.05). These results indicate that peNDF content affects digestibility and chewing activity. Consequently, the peNDF content of TMR should be considered for improving feed efficiency, digestibility, body weight gain, and performance in fattening heifers.

Anti-inflammatory effects of extract from Haliotis discus hannai fermented with Cordyceps militaris mycelia in RAW264.7 macrophages through TRIF-dependent signaling pathway.

In this study, Haliotis discus hannai (H. discus hannai) fermentation was attempted with Cordyceps militaris (C. militaris) mycelia using a solid culture. We tried to ferment H. discus hannai to determine the optimal conditions fermentation with regards to its anti-inflammatory effects. The extracts of H. discus hannai fermented with C. militaris mycelia (HFCM-5) showed higher nitric oxide inhibitory effects than H. discus hannai and C. militaris alone. HFCM-5 also decreased pro-inflammatory cytokines, TNF-α and IL-6 in a dose-dependent manner. HFCM-5 did not affect the MyD88-dependent pathway, but decreased phosphorylation of IRF3 and STAT1 which are involved in TRIF-dependent pathway. Taken together, our results suggest that HFCM-5 exerts its anti-inflammatory effects via TRIF signaling pathway and could potentially be used as a functional food in the regulation of inflammation.

Sulfated chitosan oligosaccharides suppress LPS-induced NO production via JNK and NF-κB inactivation.

Various biological effects have been reported for sulfated chitosan oligosaccharides, but the molecular mechanisms of action of their anti-inflammatory effects are still unknown. This study aimed to evaluate the anti-inflammatory effects of sulfated chitosan oligosaccharides and to elucidate the possible mechanisms of action. The results showed that pretreated low molecular weight sulfated chitosan oligosaccharides inhibited the production of nitric oxide (NO) and inflammatory cytokines such as IL-6 and TNF-α in lipopolysaccharide (LPS)-activated RAW264.7 cells. The sulfated chitosan oligosaccharides also suppressed inducible nitric oxide synthase (iNOS), phosphorylation of JNK and translocation of p65, a subunit of NF-κB, into the nucleus by inhibiting degradation of IκB-α. Our investigation suggests sulfated chitosan oligosaccharides inhibit IL-6/TNF-α in LPS-induced macrophages, regulated by mitogen-activated protein kinases (MAPKs) pathways dependent on NF-κB activation.

Enhancing Nutritional Quality of Silage by Fermentation with Lactobacillus plantarum.

The present study was aimed to investigate the nutritive profiles, microbial counts and fermentation metabolites in rye, Italian rye-grass (IRG) and barley supplemented with Lactobacillus plantarum under the field condition, and its probiotic properties. After preparation of silage, the content of crude protein (CP), crude ash, acid detergent fiber (ADF), and neutral detergent fiber (NDF), microbes such as lactic acid bacteria (LAB), yeast and fungi counts, and fermentation metabolites lactic acid, acetic acid and butyric acid was assessed. Results indicated that the content of ADF and NDF were significantly varied between rye, IRG and barley mediated silages. The content of CP was increased in L. plantarum supplemented with IRG, but slightly decreased in rye and barley mediated silages. The maximum LAB count was recorded at 53.10 × 10(7) cfu/g in rye, 16.18 × 10(7) cfu/g in IRG and 2.63 × 10(7) cfu/g in barley silages respectively. A considerable number of the yeasts were observed in the IRG silages than the rye silages (P < 0.05). The amount of lactic acid production is higher in L. plantarum supplemented silages as compared with control samples (P < 0.05). It was confirmed that higher amount of lactic acid produced only due to more number of LAB found in the silages. L. plantarum was able to survive at low pH and bile salt and the duodenum passage with the highest percentage of hydrophobicity. Furthermore, the strain was sensitive towards the antibiotics commonly used to maintain the microbes in food industrial setups. In conclusion, supplementation of L. plantarum is most beneficial in rye, IRG and barley silage preparations and probiotic characteristics of L. plantarum was an intrinsic feature for the application in the preparation of animal feeds and functional foods.

Purification of a novel nitric oxide inhibitory peptide derived from enzymatic hydrolysates of Mytilus coruscus.

Shellfish contain significant levels of high quality protein and are therefore a potential source for biofunctional high-value peptides. To purify a novel anti-inflammatory peptide from Mytilus coruscus (M. coruscus), we applied enzymatic hydrolysis and tangential flow filtration (TFF) and investigated its nitric oxide inhibitory property. To prepare the peptide, eight proteases were employed for enzymatic hydrolysis. Flavouzyme hydrolysates, which showed clearly superior nitric oxide inhibitory activity on lipopolysaccharide (LPS)-stimulated RAW264.7, were further purified using a TFF system and consecutive chromatographic methods. Finally, a novel anti-inflammatory peptide composed of 10 amino acid residues was obtained, and the sequence was identified as Gly-Val-Ser-Leu-Leu-Gln-Gln-Phe-Phe-Leu at N-terminal position. The peptide from M. coruscus effectively inhibited nitric oxide production on macrophage cells. This is the first report of an anti-inflammatory peptide derived from the hydrolysates of M. coruscus.

Purification of a novel peptide derived from Mytilus coruscus and in vitro/in vivo evaluation of its bioactive properties.

Excess oxidant can promote inflammatory responses. Moreover, chronic inflammation accompanied by oxidative stress is connected various steps involved in many diseases. From the aspect, we investigated an antioxidant peptide to prevent inflammatory response against oxidant overexpression. To prepare the peptide, eight proteases were employed for enzymatic hydrolysis, and the antioxidant properties of the hydrolysates were investigated using free radical scavenging activity by electron spin resonance (ESR) spectrometry. Papain hydrolysates, which showed clearly superior free radical scavenging activity, were further purified using consecutive chromatographic methods. Finally, a novel antioxidant peptide was obtained, and the sequence was identified as Ser-Leu-Pro-Ile-Gly-Leu-Met-Ile-Ala-Met at N-terminal. Oral administration of the peptide to mice effectively inhibited malondialdehyde (MDA) levels in a thiobarbituric acid reactive substances (TBARS) assay, and we also confirmed the antioxidative enzyme activities in superoxide dismutase (SOD) and glutathione-s-transferase (GST) assays. This is the first report of an antioxidant peptide derived from the hydrolysate of Mytilus coruscus, and also these results suggest that the peptide possesses potent antioxidant activity, and potential to enhance anti-inflammatory response.

Chitooligosaccharides induce apoptosis in human myeloid leukemia HL-60 cells.

In this study we propose a novel anticancer agent using hetero-chitooligosaccharide (hetero-COS). To examine the possibility of the hetero-COS as a anticancer agent, we prepared nine kinds of hetero-COS with relatively higher molecular weights (90, 75 and 50-COS I, 5-10kDa), medium molecular weights (90, 75 and 50-COS II, 1-5kDa), and lower molecular weights (90, 75 and 50-III, below 1kDa), and their anticancer properties were investigated on HL-60 cells using flow cytometry and morphological analysis. The results obtained indicate that 90-COS III, which is relatively higher degree of deacetylation and lower molecular weights, showed the highest anticancer activity, and the data showed the anticancer property of the hetero-COSs depended on their degree of deacetylation values and molecular weight.

Purification and characterization of a novel peptide with inhibitory effects on colitis induced mice by dextran sulfate sodium from enzymatic hydrolysates of Crassostrea gigas.

The anti-inflammatory activity of purified peptides from Crassostrea gigas (C. gigas) hydrolysates was studied. To prepare hydrolysates from C. gigas, we used eight different proteinases and the anti-inflammatory activities were determined using a nitric oxide (NO) assay in RAW264.7 cells. Among the hydrolysates, Protamex hydrolysates showed the highest anti-inflammatory activity. We separated and purified the total hydrolysate using an ultrafiltration membrane system and consecutive chromatographic methods. Finally, we obtained a peptide with the following sequence: Gln-Cys-Gln-Cys-Ala-Val-Glu-Gly-Gly-Leu at N-terminal position. The anti-inflammatory peptide purified from C. gigas inhibited NO production by 72.2% compared to the lipopolysaccharide (LPS) treated group. In addition, the Protamex hydrolysates from C. gigas showed decreased serum IgE levels and increased spleen CD4(+)/CD8(+) levels on dextran sulfate sodium (DSS) induced colitis in mice. These results suggest the peptide and hydrolysate from C. gigas possess potent anti-inflammatory effect.

Dieckol isolated from Ecklonia cava protects against high-glucose induced damage to rat insulinoma cells by reducing oxidative stress and apoptosis.

Pancreatic ß cells are very sensitive to oxidative stress and this might play an important role in ß cell death with diabetes. The protective effect of dieckol, one of the phlorotannin polyphenol compounds purified from Ecklonia cava (E. cava), against high glucose-induced oxidative stress was investigated by using rat insulinoma cells. A high-glucose (30 mM) treatment induced the death of rat insulinoma cells, but dieckol, at a concentration 17.5 or 70 µM, significantly inhibited the high-glucose induced glucotoxicity. Treatment with dieckol also dose-dependently reduced thiobarbituric acid reactive substances (TBARS), the generation of intracellular reactive oxygen species (ROS), and the nitric oxide level increased by a high glucose concentration. In addition, the dieckol treatment increased the activities of antioxidative enzymes including catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-px) in high glucose-pretreated rat insulinoma cells. Dieckol protected rat insulinoma cells damage under high glucose conditions. These effects were mediated by suppressing apoptosis and were associated with increased anti-apoptotic Bcl-2 expression, and reduced pro-apoptotic cleaved caspase-3 expression. These findings indicate that dieckol might be useful as a potential pharmaceutical agent to protect against the glucotoxicity caused by hyperglycemia-induced oxidative stress associated with diabetes.

Chitooligosaccharides decreases plasma lipid levels in healthy men.

Chitosan, which is derived from chitin, has drawn much attention due to its low toxicity and potential use in medical and pharmaceutical applications. The biological activities of chitosan have been shown to depend on its molecular weight (MW) and degree of deacetylation. In this study, we investigated whether oral chitooligosaccharides, which are easily absorbed into the body, can reduce the plasma level of lipid in smokers and non-smokers because smoking is a high-risk factor for cardiovascular diseases. All healthy men (11 smokers and 8 non-smokers) consumed 500 mg of chitooligosaccharides in water twice daily before a meal (breakfast and dinner) over a 6-week period. Total cholesterol and low-density lipoprotein cholesterol levels were significantly decreased in both the smoker group and non-smoker group when compared with baseline. These results suggest that low MW chitooligosaccharides would be an effective dietary supplement for lowering cholesterol level.

Isolation and identification of an antioxidant flavonoid compound from citrus-processing by-product.

BACKGROUND: Large amounts of citrus by-products are released from juice-processing plants every year. Most bioactive compounds are found in the peel and inner white pulp. Flavonoids are a widely distributed group of bioactive compounds. The methanolic extract of citrus peel powder has been shown to possess strong antioxidant activity. Therefore the aim of this study was to isolate the major antioxidant flavonoid compound from Citrus unshiu (satsuma) peel as citrus by-product and evaluate its antioxidant activity. RESULTS: The major flavonoid isolated from C. unshiu peel was identified as quercetagetin. The structure of the compound was determined by tandem mass spectrometry and ultraviolet spectroscopy. Its antioxidant activity was assessed by assays of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical, hydroxyl radical and intracellular reactive oxygen species (ROS) scavenging and DNA damage inhibition. Quercetagetin showed strong DPPH radical-scavenging activity (IC50 7.89 µmol L⁻¹) but much lower hydroxyl radical-scavenging activity (IC50 203.82 µmol L⁻¹). Furthermore, it significantly reduced ROS in Vero cells and showed a strong protective effect against hydrogen peroxide-induced DNA damage. CONCLUSION: The results of this study suggest that quercetagetin could be used in the functional food, cosmetic and pharmaceutical industries.