• Journal of Chest Surgery
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• v.40 no.4 s.273
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• pp.264-272
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• 2007

Background: Vascular endothelial growth factor (VEGF) plays an important role in angiogenesis, including stimulating the proliferation and migration of vascular smooth muscle cells (VSMCs). It has been known that diabetes is associated with accelerated cellular proliferation via VEGF, as compared to that under a normal glucose concentration. We investigated the effects of selective blockade of a VEGF receptor by using anti-Flt-1 peptide on the formation and hyperplasia of the neointima in balloon injured-carotid arteries of OLETF rats and also on the in vitro VSMCS' migration under high glucose conditions. Material and Method: The balloon-injury method was employed to induce neointima formation by VEGF. For f4 days beginning 2 days before the ballon injury, placebo or vascular endothelial growth factor receptor-1 (VEGFR-1) specific peptide (anti-Flt-1 peptide), was injected at a dose of 0.5mg/kg daily into the OLETF rats. At 14 days after balloon injury, the neointimal proliferation and vascular luminal stenosis were measured, and cellular proliferation was assessed by counting the proliferative cell nuclear antigen (PCNA) stained cells. To analyze the effect of VEGF and anti-Flt-1 peptide on the migration of VSMCs under a high glucose condition, transwell assay with a matrigel filter was performed. And finally, to determine the underlying mechanism of the effect of anti-Flt-1 peptide on the VEGF-induced VSMC migration in vitro, the expression of matrix metalloproteinase (MMP) was observed by performing reverse transcription-polymerase chain reaction (RT-PCR). Result: Both the neointimal area and luminal stenosis associated with neointimal proliferation were significantly decreased in the anti-Flt-1 peptide injected rats, ($0.15{\pm}0.04 mm^2$ and $ 36.03{\pm}3.78%$ compared to $0.24{\pm}0.03mm^2\;and\;61.85{\pm}5.11%$, respectively, in the placebo-injected rats (p<0.01, respectively). The ratio of PCNA(+) cells to the entire neointimal cells was also significantly decreased from $52.82{\pm}4.20%\;to\;38.11{\pm}6.89%$, by the injected anti-Flt-1 peptide (p<0.05). On the VSMC migration assay, anti-Flt-1 peptide significantly reduced the VEGF-induced VMSC migration by about 40% (p<0.01). Consistent with the effect of anti-Flt-1 peptide on VSMC migration, it also obviously attenuated the induction of the MMP-3 and MMP-9 mRNA expressions via VEGF in the VSMCS. Conclusion: Anti-Flt-1 peptide inhibits the formation and hyperplasia of the neointima in a balloon-injured carotid artery model of OLETF rats. Anti-Flt-1 peptide also inhibits the VSMCs' migration and the expressions of MMP-3 and MMP-9 mRNA induced by VEGF under a high glucose condition. Therefore, these results suggest that specific blockade of VEGFR-1 by anti-Flt-1 peptide may have therapeutic potential against the arterial stenosis of diabetes mellitus patients or that occurring under a high glucose condition.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.19 no.3
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• pp.219-226
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• 1986

To confirm the application of a newer in vitro assays to determining the nutritional value of marine crustaceans (mainly shrimps and crabs), which have been considered to be highly nutritive depending on their levels of the essential amino acids and digestibility, their C-PERs and DC-PERs were determined and studied the factors influencing their in vitro results. Four species of seawater shrimps and 2 species of seawater crabs were used in this experiment. The in vitro digestibilities showed $83{\sim}86\%$ for raw shrimps and the trypsin indigestibile substrate content (TIS) was ranged from 1.32 to 3.33 mg/g solid expressed quantitatively as mg of purified soybean trypsin inhibitor. The smaller size of shrimps revealed a greater in vitro digestibility and a lower contents of TIS. It was noted that the in vitro digestibility of raw blue crab meat was around $85\%$ while boiled tenner crab meat showed $86\%$ or above, and the leg meat had the greatest in vitro digestibility in the various parts of crab meats. The poor in vitro digestibilities for shrimp's and crab's meat, compared with that of the other seafoods as noted in previous reports, suggest that the drop in pH, due to the change in their freshness during harvesting and frozen storage, resulted in underestimating their digestibilities using four-enzyme digestion technique. The lysine contents in all samples were higher than that of ANRC casein but they contained a slightly lower sulfur-containing amino acids than those in ANRC casein. But the other EAA, such as valine, tyrosine and phenylalanine, were found to be a half as little as that in casein and played a key-factor in calculation of C-PER or DC-PER. It was observed that the value of C-PER and DC-PER for all samples ranged from 2.1 to 2.4, and the predicted digestibilities showed $90\%$ or above in all samples. It was a different results from the fact that the animal proteins bear a higher values and predicted digestibilities than those of C-PER values. The lack of correlation between C-PER and DC-PER values is attributable to the fact that the lower content of valine, tyrosine and phenylalanine, and drop in pH owing to the changes of freshness in marine crustacea proteins. Therefore, if a newer in vitro digestion technique-which are taken into account the pH drop before digestion, TIS content and released free amino acids and/or peptides-developed, C-PER assays can provide more advantages in assessing the protein nutritional value of marine crustacea than any other in vitro assays.

• Korean Journal of Food Science and Technology
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• v.26 no.3
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• pp.204-212
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• 1994

To estimate the possibility of wine-making with Korean dried persimmon, its homogenized and filtered solution was fermented at $15^{\circ}C$ and $25^{\circ}C$ for 12 weeks with Saccharomyces cerevisiae (Japan Alcoholic Beverage Association N0.7). Sugars of dried persimmon were mainly composed of 27.02% of glucose, 19.81% of fructose and 5.12% of mannose. In the fermentation at $25^{\circ}C$, glucose was almost completely consumed in 8 days, but fructose and mannose were consumed up to 64% and 74%, respectively, in the same period and were not utilized any more afterwards. In the fermentation at $15^{\circ}C$, 75% of glucose, 20% of fructose and 49% of mannose were consumed in 8 days and these sugars were continuously utilized for 12 weeks. Organic acids in the homogenized and filtered solution were levulinic acid (148.6 mg%), 4-methylvaleric acid (73.5 mg%), oxalic acid (28.7 mg%), acetic acid (8.5 mg%), N-butyric acid (8.4 mg%) and succinic acid (6.7 mg%). Irrespective of fermentation temperature, levulinic acid rapidly reduced according to progression of fermentation. Oxalic acid, N-butyric acid and succinic acid decreased at 2nd day of fermentation, and then increased at 4th and 6th days and subsequently decreased again under the levels of the solution. Acetic acid and 4-methylvaleric acid increased with the proceeding of fermentation and at 12th week of fermentation these contents were more than those of the solution. The contents of total free amino acid significantly reduced at 2th day of fermentation and then increased to the level of the solution at 12th week irrespective of fermentation temperature. Ethanol content rapidly increased to the levels of 5.3(v/v) at $15^{\circ}C$ and 9.4%(v/v) at $25^{\circ}C$ to 8th day after fermentation, but at 12th week its content was 14.5%(v/v) at $15^{\circ}C$ and 9.4%(v/v) at $25^{\circ}C$. The higher alcohots identified were 2-methyl-l-propanol, 3-methyl-ibutanol, 2-methyl-l-butanol and 2-methyl-2-propanol and the range of those contents was from 0.001% (v/v) to 0.06%(v/v). The color of the wine fermented at $15^{\circ}C$ was slightly superior but flavor and taste were slightly superior in the wine fermented at $25^{\circ}C$.

• Journal of the Korean Society of Food Science and Nutrition
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• v.36 no.8
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• pp.960-964
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• 2007

This study was performed to examine the toxicity of Psoralea corylifolia L. by the single-dose oral toxicity tests in rat and bacterial reverse mutation assay. In single-dose oral toxicity tests, 5 mL ethanol extract of P. corylifolia L. were directly injected into 10 rats (5 males and 5 females) at a dosage of 2 g/kg. Death practice was not detected during breeding periods (14 days), and $LD_{50}$ was calculated over 2 g/kg. No difference were observed with control group in the growth rate and histological observations. In bacterial reverse mutation assay, his(-) Salmonella Typhimurium TA98, TA100, TA1535, TA1537 and trp(-) Escherichia coli WP2uvrA (pKM101) were used for assessing the toxicity of ethanol extracts of P. corylifolia L.. No significant difference in formation of the colonies and no dose-dependent increase was observed regardless of the addition of S9 mix. The results showed that ethanol extracts of P. corylifolia L. did not have single-dose oral toxicity and mutagenic toxicity.

• Korean Journal of Medicinal Crop Science
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• v.10 no.3
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• pp.222-229
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• 2002

Ixeris dentata root were extracted with methanol and then fractionated with n-hexane, EtOAc and BuOH to get active fractions. and their antioxidant and antimicrobial activities in each fraction were determined. Ethyl acetate fraction of Ixeris dentata root showed strong antioxidant activities, but aqueous fraction did not show any activities. But in the antimicrobial test, aqueous fraction showed strong antimicrobial activities except to Escherichia coli. especially, aqueous fraction showed the strongest activities against Hypocrea nigricans. and butanol fraction showed the strongest activities against Cladosporium herbarum. This study was performed to determine the antimutagenic and cytotoxic effect of Ixeris dentata root methanol extract on Salmonella typhimurium TA98, TA100 and cancer cell lines using ames test and cytotoxicity assay, respectively. Cancer cell lines include human lung carcinoma(A549), human breast adenocarcinoma(MCF-7) and human hepatocellular carcinoma (Hep 3B). Futher fractionations with hexane, ethyl acetate, butanol and water from methanol extract of Ixeris dentata root were performed to obtain effective fraction, methanol extracts showed 79.94% inhibitory effect on the mutagenesis induced by N' -methyl- N' -nitro-N-nitrosoguanidine(MNNG) against TA100, while 89.99% inhibition was observed on the mutagenesis induced by 4-nitroquinoline-1-oxide(4NQO) against TA98. In the meanwhile, butanol fraction showed 89.92% and 71.01% inhibitory effect on the mutagenesis induced by benzo(a)pyrene(B(a)P) against TA98 and TA100, respectively. Ethyl acetate fraction showed the strongest effect against A549, MCF-7 and Hep3B at the same concentration compared to those of other fration.

• Applied Biological Chemistry
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• v.49 no.2
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• pp.91-96
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• 2006

Antioxidant activities and biological properties such as antimutagenic and cytotoxic effect of Phellinus linteus extracts from different extraction conditions were measured against Salmonella typhimurium and human cancer cell lines. DPPH free radical scavenging activities of the extracts were higher in the solutions extracted with ethanol (17.14) and ethanol after water (17.79), respectively. In the Ames test, ethanol extract of P. linteus alone did not exhibit any mutagenicity but showed substantial inhibitory effect against mutations induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), 4-nitroquinoline-1-oxide (4NQO), and benzo $({\alpha})$ pyrene $(B({\alpha})P)$. The extracts of ethanol and ethanol after water of P. linteus $(200\;{\mu}g/plate)$ had the highest inhibitory effect of 61.5 and 60.9%, respectively, on the mutagenesis on S. typhimurium TA98 strain induced by $B({\alpha})P$. Extracted solutions of ethanol and ethanol after water of P. linteus showed high antimutagenic effect against MNNG, 4NQO, Trp-P-1 and $B({\alpha})P$, causing mutations in S. typhimurium TA100 strain. The anticancer effects of P. linteus extracts were investigated against human fibrosarcoma HT-29 and human hepatocellular carcinoma HepG2. The treatment of 0.5 mg/ml of ethanol, ethanol after water and water extracts of P. linteus had the highest cytotoxicity of 59, 57, 54%, respectively against HT-27 cell line, whereas low cytotoxicity effects were observed against HepG2 cell line in the range of $10{\sim}30%$. The ethanol and water extracts of P. linteus also showed the nitrate scavenging ability at different pHs. The ethanol extract showed higher nitrate-scavenging ability compared to water extract of P. linteus.

• Korean Journal of Medicinal Crop Science
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• v.10 no.2
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• pp.132-138
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• 2002

This study was performed to determine the antimutagenic and cytotoxic effect of ,Kalopanacis cortex endothelium. Methanol extract was used on Salmonella typhimurium TA98, TA100 and three cancer cell lines. In the Ames test, methanol extract of Kalopanacis cortex endothelium alone did not exhibit any mutagenicity but showed substantial inhibitory effects against mutation induced by 4-nitroquinoline-l-oxide(4NQO), N'-methyl- N'-nitro-N-nitrosoguanidine(MNNG) and benzo(a)pyrene(B(a)P). The methanol extract of Kalopanacis cortex endothelium showed approximately 79.29% and 75.38% inhibitory effect on the mutagenesis induced by 4NQO and B(a)P, respectively, against TA98 strain. Whereas 79.49%, 89.3% and 68.85% inhibitions were observed on the mutagenesis induced by 4NQO, MNNG and B(a)P against TA100 strain. Methanol extracts from Kalopanacis cortex endothelium showed high antimutagenic effects against 4NQO, MNNG and B(a)P. The anticancer effects of methanol extract from Kalopanacis cortex endothelium against human lung carcinoma(A549), human breast adenocarcinoma (MCF-7) and human hepatocellular carcinoma(Hep3B) were investigated. The treatment of 0.5mg/ml Kalopanacis cortex endothelium methanol extracts had the highest cytotoxicity against A549(81.54%), followed by MCF-7(81.92%) and Hep3B (78.57%). In contrast 0.5mg/ml treatment of methanol extracts from Kalopanacis cortex endothelium had only $4{\sim}25%$ cytotoxicity on normal human liver cell(293).