A chemometric study on the identification of 5-methylfurfural and 2-acetylfuran as particular volatile compounds of oxidized fish oil based on SHS-GC-IMS.

Fish oil develops particular off-odors, mainly fishy odor, from the oxidation of its characteristic fatty acids, docosahexaenoic (DHA) and eicosapentaenoic (EPA). Anchovy oil (AO) was taken as representative of fish oils. This was compared to three vegetable oils with different fatty acid compositions, i.e. camellia, sunflower and linseed oil, and differential volatile compounds were identified by static-headspace gas-chromatography ion-mobility-spectrometry (SHS-GC-IMS) and orthogonal partial-least-squares discriminant analysis (OPLS-DA) during oxidation at 60 °C. Three groups of differential volatile compounds detected at higher concentrations in the AO were screened out and two compounds, identified as 5-methylfurfural and 2-acetylfuran, were characteristic to the AO and not found in the vegetable oils. They were formed from both EPA and DHA, only present in the AO, and their formation mechanisms were proposed. The contents of 5-methylfurfural and 2-acetylfuran increased linearly with the oxidation time and consequently they could be used as oxidative markers of fish oils.

Concomitant oxidation of fatty acids other than DHA and EPA plays a role in the characteristic off-odor of fish oil.

The aim of the present research was to explore the development of off-odors in fish oil from the perspective of fatty acid oxidation. It was found that the off-odors elicited by the two major ω-3 PUFAs in fish oil, i.e. DHA and EPA, were different from those by fish oil. Results showed that simultaneous oxidation of fatty acids other than DHA and EPA can be involved. The off-odors of fish oil was successfully simulated by combining oxidized samples of DHA, EPA and sunflower oil. Therefore, oxidation of oleic and linoleic acids also contributed to the off-odors in fish oil. A novel analytical approach that consisted in the combination of gas chromatography-ion mobility spectrometry (GC-IMS) and orthogonal partial least squares discriminant analysis (OPLS-DA) was applied to identify differences in the volatile components between the recombinant oil and the fish oil.

Driver mutations in ADGRL3 are involved in the evolution of ependymoma.

Although there have been recent advances in the molecular pathology of ependymomas, little is known about the underlying molecular evolution during its development. Here, we assessed the clinical, pathological and molecular evolutionary process of ependymoma recurrence in a 9-year-old patient who had seven recurrences of supratentorial ependymoma and died from intracranial multiregional recurrences at the age of 19 years old. Whole-genome sequencing (WGS) of 7 tumor samples (1 primary and 6 subsequent recurrent tumors) was performed to elucidate the mutation landscape and identify potential driver mutations for tumor evolution. The genetic profiles of the seven tumor specimens showed significant heterogeneity and suggested a highly branched evolutionary pattern. The mutational signatures and chromothripsis changed with treatments. Strikingly, adhesion G protein-coupled receptor L3 (ADGRL3, also known as Latrophilins 3, LPNH3) was found to be consistently mutated during the entire disease process. However, Sanger sequencing of other 78 ependymoma patients who underwent surgery at our institution showed no genetic alteration of ADGRL3, as found in the present case. The mRNA levels of ADGRL3 were significantly lower in ependymomas (n = 36), as compared with normal brain tissue (n = 3). Grade III ependymomas had the lowest ADGRL3 expression. Moreover, ependymomas with lower mRNA level of ADGRL3 had shorter overall survival. Our findings, therefore, demonstrate a rare evolutionary process of ependymoma involving ADGRL3.

Sea Cucumber Sterol Alleviates the Lipid Accumulation in High-Fat-Fructose Diet Fed Mice.

The effect of marine-derived sea cucumber sterol (SS) with a special sulfate group on lipid accumulation remains unknown, although phytosterol has been proved to have many biological activities, including lowering blood cholesterol. The purpose of the present study is to investigate the alleviation of SS on lipid accumulation and the possible underlying mechanism using high-fat-fructose diet fed mice. Dietary administration with SS for 8 weeks reduced significantly the body weight gain and lipid levels in serum and liver. Especially, SS was superior to phytosterol in lowering lipid accumulation due to the great promotion of fatty acid ß-oxidation, the inhibition of cholesterol synthesis, and the acceleration of cholesterol efflux. The findings found that sea cucumber sterol exhibited a more significant effect than phytosterol on alleviating HFF-diet-induced lipid accumulation through regulating lipid and cholesterol metabolism, which might be attributed to the difference in the branch chain and sulfate group.

Characterization and Absorption Kinetics of a Novel Multifunctional Nanoliposome Stabilized by Sea Cucumber Saponins Instead of Cholesterol.

Cholesterol was usually used to stabilize liposome, although there have been controversies on the relationship between dietary cholesterol and health. The present study aimed to prepare a novel multifunctional nanoliposomes stabilized by sea cucumber-derived saponins using ultrasound-assisted film dispersion method. A novel uniform liposome with a mass ratio of egg yolk lecithin/sea cucumber saponins at 75:25 was successfully prepared to encapsulate saponin, and the particle size was 164.8 ± 1.70 nm with a PDI value of 0.214 ± 0.022 and zeta potential of -15.97 ± 1.23 mV. The digestion and absorption results in vivo showed that the dietary saponins in liposome form could delay the peak time of saponins and prolong their residence time in the serum. Moreover, saponins were more easily converted into their corresponding metabolites after administration with saponins in the liposome form. The novel liposome as an efficient carrier with multiple functions had great potential in the development of functional food and biomedical applications.

Application of Plackett-Burman Design in Screening of Natural Antioxidants Suitable for Anchovy Oil.

Considering the safety of synthetic antioxidants, more and more natural antioxidants have been developed and utilized in foods. This study aimed to screen out a natural antioxidant combination from many antioxidants, which could significantly affect the oxidation stability of anchovy oil, while Plackett-Burman design (PBD) methodology was employed in this screening. According to the statistical results of this design, sesamol, dihydromyricetin, teapolyphenol, and rosemary acid were four significant parameters on the oxidation stability of anchovy oil. Moreover, dihydromyricetin presented the best antioxidant effect among nine kinds of selected antioxidants when they were used alone in anchovy oil. Meanwhile, a combination including sesamol (0.02%), teapolyphenol (0.02%). and rosemary acid (0.02%) was adopted, and its antioxidant ability was similar to that of tert-butylhydroquinone (TBHQ). Additionally, phytic acid as a synergist was used and combined with sesamol, and the antioxidant ability of this combination was better than that of TBHQ. This study presented a reference for the industrial applications of natural antioxidants and synergists in anchovy oil.

A Comprehensive Review of Spirit Drink Safety Standards and Regulations from an International Perspective.

Standards and regulations related to spirit drinks have been established by different countries and international organizations to ensure the safety and quality of spirits. Here, we introduce the principles of food safety and quality standards for alcoholic beverages and then compare the key indicators used in the distinct standards of the Codex Alimentarius Commission, the European Union, the People's Republic of China, the United States, Canada, and Australia. We also discuss in detail the "maximum level" of the following main contaminants of spirit drinks: methanol, higher alcohols, ethyl carbamate, hydrocyanic acid, heavy metals, mycotoxins, phthalates, and aldehydes. Furthermore, the control measures used for potential hazards are introduced. Harmonization of the current requirements based on comprehensive scope analysis and the risk assessment approach will enhance both the trade and quality of distilled spirits. This review article provides valuable information that will enable producers, traders, governments, and researchers to increase their knowledge of spirit drink safety requirements, control measures, and research trends.

Effects of different fatty acids composition of phosphatidylcholine on brain function of dementia mice induced by scopolamine.

BACKGROUND: Phosphatidylcholine (PC), the major source of dietary choline, has been demonstrated to improve the capability of learning and memory in rodent and the amelioration of long-chain n-3 polyunsaturated fatty acids (PUFA) on anti-aging and anti-oxidation is widely known as well. In this study, three kinds of PC were chose to demonstrate the role of different fatty acids composition on glycerol backbone in improving the brain function of mice induced by scopolamine which was used to impair cholinergic system and cause oxidative stress. METHODS: Male BALB/c mice were randomly divided into 5 groups: model (M) group, control (Con) group, egg yolk lecithin (EL) group, squid PC (SQ-PC) group and sea cucumber PC (SC-PC) group. The intraperitoneal injection of scopolamine hydrobromide (5 mg/kg) was carried out on the 8(th) of group feeding and sustained daily until the end of test. Morris water maze test was used to evaluate the improvement of cognitive decline and the activity of acetylcholinesterase (AchE), superoxide dismutase (SOD) and monoamine oxidase (MAO) and malondialdehyde (MDA) content in brain were measured to assess the physiological changes. RESULTS: In behavior test, the latency of PC groups was significantly reduced, while number of crossing the platform and time in target quadrant were increased in comparison with M group and the improvements of SQ-PC and SC-PC were better than that of EL (P < 0.05). Similar trend was observed in physiological changes. The AchE activity was effectively decreased and the SOD activity increased in hippocampus, cortex and white matter when comparing PC groups with M group. SQ-PC, SC-PC and EL respectively showed 22.82, 28.80 and 11.81 % decrease in MDA level in brain compared with M group. The MAO activity in white matter of SQ-PC, SC-PC and EL group separately depressed 33.05, 33.64 and 19.73 % in comparison with M group. No significance between SQ-PC and SC-PC was found in these indicators except the SOD activity in hippocampus and white matter. SQ-PC group had a higher SOD activity in hippocampus (103.68U/mg · prot.) and lower in white matter (120.57 U/mg · prot.) than SC-PC group (95.53 U/mg · prot. in hippocampus, 134.49 U/mg · prot. in white matter). PC rich in n-3 PUFA acted more ameliorative effects than that barely contained on the indicators above. CONCLUSIONS: Different fatty acids composition of PC all could diminish the cognitive decline and biological damage and protect the brain. EPA and DHA partly enhaced to the advantageous effects.

Molecular species analysis of monosialogangliosides from sea urchin Strongylocentrotus nudus by RPLC-ESI-MS/MS.

Sea urchin gangliosides have been proved to contain neuritogenic activities, which related to their molecular compositions. This study reports a method utilizing reversed-phase chromatography coupled to mass spectrometry for structure investigation and molecular species determination of the monosialogangliosides from sea urchin Strongylocentrotus nudus. Two types of sulfated and nonsulfated monosialogangliosides were isolated from the sea urchin ovary. In MS(2) spectra of both nonsulfated monosialoganglioside and sulfated monosialoganglioside, 2-6 linked sialic acids were identified by the characteristic fragments of (0,4)A2-CO2 and (0,2)A1. Fragment ions at m/z 139.1 and m/z 169.1 of nonsulfated monosialoganglioside might be characteristic for 8-sulfated sialic acid residue. Retention time of the molecules was effectively used in the characterization of unknown molecules, and molecules that differ in mass by only 0.04 Da were easily differentiated.

Fucosylated chondroitin sulfate from sea cucumber improves glucose metabolism and activates insulin signaling in the liver of insulin-resistant mice.

This study investigated the effects of fucosylated chondroitin sulfate (CHS) isolated from sea cucumber on glucose metabolism and insulin signaling in the liver of insulin-resistant C57BL/6 mice fed a high-fat, high-sucrose diet (HFSD). Male C57BL/6J mice were randomly assigned into six groups: control; HFSD; 1 mg RSG/kg·body weight (RSG); 80 mg CHS/kg · body weight (CHS); 20 mg CHS+1 mg RSG/kg · body weight (20 CHS+RSG); and 80 mg CHS+1 mg RSG/kg · body weight (80 CHS+RSG). Blood glucose, insulin parameters, glucose metabolism-related enzymes activities and insulin-signaling transducers in the liver were analyzed at 19 weeks. Results showed that CHS significantly decreased body weight gain, adipose tissue weight, and fasting blood glucose and serum insulin levels in insulin-resistant mice. Rosiglitazone (RSG) is an effective thiazolidinedione hypoglycemic agent, and CHS synergistically enhanced the effect of RSG. CHS feeding normalized the activities of hexokinase, pyruvate kinase, glycogen phosphorylase, glucose-6-phosphatase, and increased glycogen reserves in the liver. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis showed that CHS promoted the mRNA expression of insulin receptors (IR), insulin receptor substrate 2 (IRS-2), phosphatidylinositol 3 kinase (PI3K), protein kinase B (PKB), and glycogen synthase (GS) in the liver of insulin resistant mice, and inhibited glycogen synthase kinase-3 (GSK-3ß) mRNA expression. The results suggested that CHS treatment improved glucose metabolism by modulating metabolic enzymes and promoting the PI3K/PKB/GSK-3ß signaling pathway mediated by insulin at the transcriptional level. These results provided strong justification for the development of CHS as a functional food.

Effects of bioactive components of sea cucumber on the serum, liver lipid profile and lipid absorption.

Several studies had indicated that the whole body of sea cucumber had beneficial effects on lipid metabolism. However, little information has been known on the individual functions of its bioactive components, and this study was undertaken to compare the different effects on improving lipid metabolism. The rats were assigned to seven groups: control, whole sea cucumber, saponins, polysaccharides, collagen peptides, dregs and non-saponin residues. After 28 d of feeding, the serum total cholesterol, triglyceride, high-density lipoprotein-cholesterol, and hepatic lipid concentrations were examined. The results indicated that a dietary saponin supplement significantly suppressed adipose accumulation, and reduced serum and hepatic lipids. Saponin proved to be more effective than the other isolated components, so is considered to be the main lipid-lowering component in sea cucumber. The possible mechanism by which saponins improved lipid metabolism was also investigated. The saponins of sea cucumber suppressed and delayed TG and TC absorption which could be related to the pancreatic lipase inhibiting effect of saponins. This may be an important mechanism to explain its lipid-lowering effect on rats.

Short term effects of different omega-3 fatty acid formulation on lipid metabolism in mice fed high or low fat diet.

BACKGROUND: Bioactivities of Docosahexaenoic acid (DHA) and Eicosapentaenoic acid (EPA) depend on their chemical forms. The present study was to investigate short term effects of triglyceride (TG), ethyl ester (EE), free fatty acid (FFA) and phospholipid (PL) forms of omega-3 fatty acid (FA) on lipid metabolism in mice, fed high fat or low fat diet. METHOD: Male Balb/c mice were fed with 0.7% different Omega-3 fatty acid formulation: DHA bound free fatty acid (DHA-FFA), DHA bound triglyceride (DHA-TG), DHA bound ethyl ester (DHA-EE) and DHA bound phospholipid (DHA-PL) for 1 week, with dietary fat levels at 5% and 22.5%. Serum and hepatic lipid concentrations were analyzed, as well as the fatty acid composition of liver and brain. RESULT: At low fat level, serum total cholesterol (TC) level in mice fed diets with DHA-FFA, DHA-EE and DHA-PL were significantly lower than that in the control group (P < 0.05). Hepatic TG level decreased significantly in mice fed diets with DHA-TG (P < 0.05), DHA-EE (P < 0.05) and DHA-PL (P < 0.05), while TC level in liver was significantly lower in mice fed diets with TG and EE compared with the control group (P < 0.05). At high fat level, mice fed diets with DHA-EE and DHA-PL had significantly lower hepatic TC level compared with the control diet (P < 0.05). Hepatic PL concentration experienced a significant increase in mice fed the diet with PL at high fat level (P < 0.05). Furthermore, both at low and high fat levels, hepatic DHA level significantly increased and AA level significantly decreased in all forms of DHA groups (P < 0.05), compared to control groups at two different fat levels, respectively. Additionally, cerebral DHA level in mice fed diets with DHA-FFA, DHA-EE and DHA-PL significantly increased compared with the control at high fat level (P < 0.05), but no significant differences were observed among dietary treatments for mice fed diets with low fat level. CONCLUSION: The present study suggested that not only total dietary fat content but also the molecular forms of omega-3 fatty acids contributed to lipid metabolism in mice. DHA-PL showed effective bioactivity in decreasing hepatic and serum TC, TG levels and increasing omega-3 concentration in liver and brain.

The anti-tumor activities of cerebrosides derived from sea cucumber Acaudina molpadioides and starfish Asterias amurensis in vitro and in vivo.

The present study was undertaken to examine the effect of cerebrosides derived from the sea cucumber Acaudina molpadioides and the starfish Asterias amurensis on the anti-tumor activity in vitro and in vivo. The results indicated that both Acaudina molpadioides cerebrosides (AMC) and Asterias amurensis cerebrosides (AAC) exhibited an inhibitory effect on cell proliferation through induction of apoptosis in S180 cells. Moreover, administration of AMC and AAC (50 mg/kg BW) on S180 tumor bearing mice reduced the tumor weight by 45.24 % and 35.71 %, respectively. In S180 ascites tumor model, AMC and AAC (50 mg/kg BW) treatment exhibited a significant ascites fluid growth inhibition of 31.23 % and 22.72 %. Furthermore, the ascites tumor cell viability ratio in AMC and AAC groups reduced to 50.89 % and 51.69 %, respectively. The life span of AMC and AAC administrated groups increased by 55.28 % and 35.77 % compared to control. Quantitative real-time PCR analysis demonstrated that the administration of AMC and AAC down-regulated the expression of Bcl-2, Bcl-xL, while on the other hand, up-regulated Bax, Cytochrome c, caspase-9 and caspase-3 mRNA level of the S180 ascites tumor cells. It was concluded that AMC and AAC should have potential anti-tumor activity both in vitro and in vivo by inducing apoptosis through the mitochondria-mediated apoptosis pathway. AAC seemed to be more effective than AMC in vitro but less potent in vivo. It may depend on the structural differences in their fatty acid groups and sphingoid bases.

[Neuroendoscopic treatment for 372 cases of intracranial lesions].

OBJECTIVE: To explore the application of neuroendoscopic treatment for intracranial lesions. METHODS: The clinic data of 372 patients with intracranial lesions, who underwent neuroendoscopic treatment at our department from May 1998 to May 2010, were reviewed retrospectively. Representative endoscopic treatments included endoscopic third ventriculostomy (ETV) (n = 198), ETV & endoscopic biopsy (n = 69), neuroendoscopic ostomy for septum pellucidum fenestration (n = 55) (for septum pellucidum cysts, n = 37) and endoscopic cystoventriculostomy for ventricular cysts (n = 50). Their surgical indications and clinical outcomes were summarized for analysis. RESULTS: ETV was performed successfully in 369 cases. Among them, 2 failed cases underwent other operations and endoscopic biopsy failed in 1 case. Within a short post-operative period, the symptoms were resolved in 347 cases (93.3%), showed no improvement in 23 cases (6.2%) and 2 died (0.5%). At Month 6 post-operation, a failure of ETV was detected in 22 cases (9.5%), a failure of neuroendoscopic ostomy for septum pellucidum cysts in 23 (69.7%) and for ventricular cysts in 12 cases (26.7%). CONCLUSION: ETV is effective in the treatment of obstructive hydrocephalus, but its indication should be strictly controlled for children. Effective rate of neuroendoscopic treatment for intracranial septum pellucidum cysts remains unsatisfactory so that its operative indication should be strictly controlled.

Study on possible mechanism of orotic acid-induced fatty liver in rats.

OBJECTIVE: The aim of this study was to investigate the possible mechanism of orotic acid-induced fatty liver in rats. METHODS: Rats were randomly divided into two groups and fed an AIN-93 diet with 1% orotic acid or without orotic acid for 10 d. Hepatic lipid concentrations, such as triacylglycerol, total cholesterol, and phospholipids, were examined. To clarify the mechanism of orotic acid-induced fatty liver, hepatic enzyme activities and mRNA levels of key enzymes related in lipid metabolism and hepatic gene expression of transcription factors were determined. RESULTS: Orotic acid administration significantly increased hepatic triacylglycerol concentration. The activity and mRNA level of fatty acid synthase were obviously upregulated by orotic acid treatment, whereas the activities and mRNA concentrations of carnitine palmitoyl transferase and microsomal triacylglycerol transfer protein were significantly depressed. Furthermore, orotic acid stimulated the mRNA expression of sterol regulatory element binding protein-1c but did not alter the mRNA concentration of peroxisome proliferator-activated receptor-α in the liver. CONCLUSION: The stimulation of triacylglycerol synthesis induced by orotic acid is mainly caused by enhancement of sterol regulatory element binding protein-1c and its target gene involved in fatty acid biosynthesis. In contrast, the inhibition of fatty acid ß-oxidation and very-low-density lipoprotein secretion were related to the observed lipid accumulation.

[Principal component analysis and cluster analysis of inorganic elements in sea cucumber Apostichopus japonicus].

The present study is to investigate the feasibility of multi-elements analysis in determination of the geographical origin of sea cucumber Apostichopus japonicus, and to make choice of the effective tracers in sea cucumber Apostichopus japonicus geographical origin assessment. The content of the elements such as Al, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, As, Se, Mo, Cd, Hg and Pb in sea cucumber Apostichopus japonicus samples from seven places of geographical origin were determined by means of ICP-MS. The results were used for the development of elements database. Cluster analysis(CA) and principal component analysis (PCA) were applied to differentiate the sea cucumber Apostichopus japonicus geographical origin. Three principal components which accounted for over 89% of the total variance were extracted from the standardized data. The results of Q-type cluster analysis showed that the 26 samples could be clustered reasonably into five groups, the classification results were significantly associated with the marine distribution of the sea cucumber Apostichopus japonicus samples. The CA and PCA were the effective methods for elements analysis of sea cucumber Apostichopus japonicus samples. The content of the mineral elements in sea cucumber Apostichopus japonicus samples was good chemical descriptors for differentiating their geographical origins.

Dietary saponins of sea cucumber alleviate orotic acid-induced fatty liver in rats via PPARalpha and SREBP-1c signaling.

BACKGROUND: Nonalcoholic fatty liver disease is the most common chronic liver disease in the world, and is becoming increasingly prevalent. Saponins of sea cucumber (SSC) are proven to exhibit various biological activities. Therefore, the present study was undertaken to examine the effect of saponins extracted from sea cucumber (Pearsonothuria graeffei) on the preventive activity of fatty liver in rats. METHODS: Male Wistar rats were randomly divided into five groups, including normal control group, fatty liver model group, SSC-treated group with SSC at levels of 0.01%, 0.03% and 0.05%. Model rats were established by administration with 1% orotic acid (OA). After the experiment period, serum total cholesterol (TC), triglyceride (TG), and hepatic lipid concentrations were determined. To search for a possible mechanism, we examined the changes of key enzymes and transcriptional factors involved in hepatic lipids biosynthesis, fatty acid beta-oxidation. RESULTS: Both 0.03% and 0.05% SSC treatment alleviated hepatic steatosis and reduced serum TG and TC concentration significantly in OA fed rats. Hepatic lipogenic enzymes, such as fatty acid synthase (FAS), malic enzyme (ME), and glucose-6-phosphate dehydrogenase (G6PDH) activities were inhibited by SSC treatment. SSC also decreased the gene expression of FAS, ME, G6PDH and sterol-regulatory element binding protein (SREBP-1c). Otherwise, the rats feeding with SSC showed increased carnitine palmitoyl transferase (CPT) activity in the liver. Hepatic peroxisome proliferator-activated receptor (PPARalpha), together with its target gene CPT and acyl-CoA oxidase (ACO) mRNA expression were also upregulated by SSC. CONCLUSIONS: According to our study, the lipids-lowering effect of dietary SSC may be partly associated with the enhancement of beta-oxidation via PPARalpha activation. In addition, the inhibited SREBP-1c- mediated lipogenesis caused by SSC may also contribute to alleviating fatty liver.

The mechanism of dietary cholesterol effects on lipids metabolism in rats.

BACKGROUND: Cholesterol administration has been reported to influence hepatic lipid metabolism in rats. In the present study, the effect of dietary cholesterol on hepatic activity and mRNA expression of the enzymes involved in lipid metabolism were investigated. Fourteen male Wistar rats were randomly divided into 2 groups and fed 1% cholesterol or cholesterol free AIN76 diets for 4 weeks. RESULTS: The serum triglyceride and high density lipoprotein cholesterol levels were significantly decreased but the total cholesterol and non high density lipoprotein cholesterol levels were significantly increased in the cholesterol-fed rats compared with the control rats. And the concentrations of the hepatic total cholesterol and triglyceride increased about 4-fold and 20-fold separately by dietary cholesterol. The activities of hepatic malic enzyme, glucose-6-phosphate dehydrogenase, fatty acid synthase, phosphatidate phophatase and carnitine palmitoyl transferase were depressed by the cholesterol feeding (40%, 70%, 50%, 15% and 25% respectively). The results of mRNA expression showed that fatty acid synthase, carnitine palmitoyl transferase 1, carnitine palmitoyl transferase 2, and HMG-CoA reductase were down-regulated (35%, 30%, 50% and 25% respectively) and acyl-CoA: cholesterol acyltransferase and cholesterol 7alpha-hydroxylase were up regulated (1.6 and 6.5 folds) in liver by the cholesterol administration. CONCLUSIONS: The dietary cholesterol increased the triglyceride accumulation in liver, but did not stimulate the activity and the gene expression of hepatic enzymes related to triglyceride and fatty acid biosynthesis.

Differential effects of sulfated triterpene glycosides, holothurin A1, and 24-dehydroechinoside A, on antimetastasic activity via regulation of the MMP-9 signal pathway.

Two sulfated triterpene glycosides, holothurin A(1) (HA(1)) and 24-dehydroechinoside A (DHEA), isolated from the sea cucumber Pearsonothuria graeffei, are of the holostane type with 18(20)-lactone and identical carbohydrate chains. DHEA has a side chain 23 (24)-double bond, while HA(1) has a hydroxyl group at C-21. In this study, we compared the effects of DHEA and HA(1) on metastasis in vitro and in vivo. The results show that HA(1) and DHEA treatment significantly suppressed adhesion of human hepatocellular liver carcinoma cells (HepG2) to both matrigel and human endothelial cells (ECV-304) and inhibited HepG2 cell migration and invasion in a dose-dependant manner. HA(1) and DHEA reduced tube formation of ECV-304 cells on the matrigel in vitro and attenuated neovascularization in the chick embryo using the chorioallantoic membrane (CAM) assay in vivo. Immunocytochemistry analyses revealed that both HA(1) and DHEA significantly decreased the expression of the matrix metallo-proteinase-9 (MMP-9) and increased the expression level of tissue inhibitor of metalloproteinase-1 (TIMP-1), an important regulator of MMP-9 activation. Western blot analyses demonstrated that HA(1) and DHEA remarkably abolished the expression of vascular endothelial growth factor (VEGF). The expression of nuclear factor-kappa B (NF-κB) was significantly decreased by HA(1), while DHEA treatment had no effect on the down regulation of NF-κB expression. These data suggest that both DHEA and HA(1) exert significant antimetastatic activities by inhibiting MMP-9 and VEGF expression. DHEA-induced antimetastasis was more potent than HA(1). In addition, only HA(1) treatment downregulated the expression level of NF-κB, suggesting that the antimetastatic activity of triterpene glycosides derived from P. graeffei can be either NF-κB-dependent or -independent, depending on their structure.

Characterization of a pattern recognition molecule vitellogenin from carp (Cyprinus carpio).

Pattern recognition proteins function in innate immune responses by binding to molecules on the surface of invading pathogens and initiating host defense reactions. To explore the role of vitellogenin (Vg) in fish innate immunity, we purified Vg from Carp by gel filtration combined with diethylaminoethyl (DEAE) chromatography. The purified Vg was confirmed by MALDI-TOF mass spectrometry. Antibacterial activity analysis showed that Vg inhibited bacterial activity to Escherichia coli and Staphylococcus aureus in a dose-dependent manner. Vg bound to the surface of Gram-negative and Gram-positive bacteria. It also agglutinated E. coli and S. aureus and weakly to Saccharomyces cerevisiae. Vg showed a strong binding activity to lipopolysaccharides from Gram-negative bacteria. Vg-treated macrophage enhanced phagocytosis to E. coli and S. aureus. Vg also bind with macrophage function as opsonins to promote phagocytosis. The results suggest that Vg serves as a pattern recognition molecule and opsonins in antibacterial defense and as an effector in fish innate immunity.