• YAKHAK HOEJI
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• v.41 no.3
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• pp.370-380
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• 1997

The role of nitric oxide (NO) on the non-adrenergic non-cholinergic (NANC) relaxations induced by the short and prolonged electrical field stimulation (EFS) has been studied in the rabbit corpus cavernosum. In the presence of atropine and guanethidine the prolonged EFS (2-16 Hz) of corpus cavernosal strips precontracted with phenylephrine produced frequency-dependent relaxations, which were abolished by tetrodotoxin as shown in the relaxations induced gy the short EFS, indicating that their orgin is NANC nerve stimulation. $N^G$-nitro-L-arginine (L-NNA), inhibitor of nitirc oxide synthase, caused a concentration-dependent inhibition to the NANC relaxation, and at 100 M L-NNA the relaxation were virtually abolished. The inhibitory effect of L-NNA was reversed by L-arginine. Hemoglobin abolished the relaxations to NO and also caused a concentration-dependent inhibition of the NANC relaxation. The hemoglobin-resistant relaxation induced by EFS was eliminated by L-NNA. Methylene blue significantly reduced the NANC relaxation in a conentration-dependent manner. The NANC relaxation was not affected by a VIP-inactivating pepridase, alpha0chymotrypsin, whereas VIP-induced relaxation was completely abolished. NO- and VIP-induced relaxation were not affected by L-NNA. These results indicate that the NANC relaxation induced by prolonged EFS of the rabbit corpus cavernosum is mediated by NO-guanosine 3',5'-cyclic monophosphate pathway as shown in the relaxation induced by the short EFS, and that VIP release is not essential for the NANC relaxation of the rabbit corpus cavernosum and VIP is not involved the generation fo NO.

• The Korean Journal of Food And Nutrition
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• v.22 no.1
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• pp.20-28
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• 2009

According to traditional oriental medicine, only non-coagulated native deer blood is said to be effective, and coagulated deer blood is ineffective. Thus, a drying and tablet-producing method for deer blood was developed to maintain its physiological and therapeutic activity, and so that after drying, it can be redissolved and protected from coagulation. Proteases such as trypsin, pepsin, chymotrypsin, and aminopeptidase were added to the deer blood indicating that it coagulated in an hour, as shown by the reference. Wax gourd extract, which is high in protease, was added to the blood resulting in anticoagulation for 31 hours. Also, additions of 1% EDTA, 0.38% sodium citrate, 0.16% calcium oxalate, 1.2% ethanol, and 0.006% heparin to the deer blood resulted in anticoagulation for 1 hour, 4 hours, 2 hours, 1 hour, and 31 hours, respectively. In an experiment using 0.19% sodium citrate plus 1% wax gourd extract, and 0.006% heparin plus 1% wax gourd extract, anticoagulation was maintained for up to 72 hours. However, since heparin can not be used in food, the deer blood tablet was made with the addition of 0.19% sodium citrate and 1% wax gourd extract, followed by freeze drying. The dissolution rate for the tablet manufactured in this manner was 96.7%. And the dissolution rates for spray-dried deer blood, vacuum-dried deer blood, and marketed deer blood tablets were 85%, 81%, and 25.5%, respectively. The composition of the tablet produced from the freeze-dried deer blood was 56.4% protein, 18.7% lactose, 1.2% amino acids, 1.0% glucose, 0.7% lipids, 180 mg/100 g of iron, 13 mg/100 g of potassium, 39.1 mg/100 g of calcium, 480 mg/100 g of sodium, 368 mg/100 g of chloride, each.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.6
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• pp.841-847
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• 2005

The objective of this experiment was to investigate the development of gastrointestinal tract and activities of pancreatic enzymes in White Roman geese. Thirty developing embryos at the 22th, 24th and 26th day of incubation and at hatching, and sixteen or eight goslings, half males and half females, at the 1, 3, 7 or 11, 14, 21 and 28 days of age were sampled, respectively. The weights of the yolk, gastrointestinal tract and intestinal length, and the activities of pancreatic enzymes were measured. Residual yolk weight decreased rapidly during late incubation and was nearly depleted at 3 days of age. The protein and energy contents in the residual yolk of goslings at 3 days of age were significantly (p<0.05) less than those at the late incubation. From 6 days before hatching to 28 days of age, the absolute weights of gizzard, proventriculus, liver, pancreas, small intestine and large intestine in goslings increased by 48, 457, 94, 2334, 89 and 76 times, respectively. The relative weights of proventriculus, gizzard, liver, pancreas, small intestine and large intestine reached peaks at 3, 3, 14, 14, 11 and 11 days of age, respectively, and then decreased gradually. However, the relative lengths of small intestine and large intestine reached peaks at 3 days of age and at hatching, respectively. The activities of pancreatic trypsin and chymotrypsin increased sharply from hatching to 14 day of age, and then decreased gradually until 21 days of age. The activity and specific activity of pancreatic amylase were increased following by age and peaked at 7 to 11 and 21 days of age, respectively. The activity and specific activity of pancreatic lipase reached a plateau from 11 to 28 days of age. These results indicate that the gastrointestinal tract and activities of pancreatic enzymes developed more rapidly than body weight through the early growing period of goslings.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.51 no.2
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• pp.135-141
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• 2018

This study investigated the immuno-potentiating activities of Japanese mud shrimp Upogebia major. We examined the effects of enzymatic hydrolysate from U. major on the production of nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) and on the expression of pro-inflammation cytokines including $TNF-{\alpha}$, IL-6 and $IL-1{\beta}$ in RAW 264.7 cells. The treatment of six enzymatic hydrolysates of U. major (alcalase, ${\alpha}$-chymotrypsin [${\alpha}-Chy$], trypsin, pepsin, neutrase, protamex) significantly increased the production of NO in RAW 264.7 cells, with ${\alpha}-Chy$ having the greatest effect. This hydrolysate was fractionated by two ultrafiltration membranes at 3 and 10 kDa to created three fractions (below 3 kDa, between 3 and 10 kDa, and above 10 kDa). Of these, the <3 kDa and >10 kDa fractions showed significant increases in NO production. These two fractions also induced $PGE_2$ production in RAW 264.7 cells and showed significant increases in the expression of all cytokines studied. These results suggest that enzymatic hydrolysate from U. major is a potentially useful food material with immune-potentiating effects.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.53 no.5
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• pp.707-716
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• 2020

Red sea cucumber Stichopus japonicus, was dried using three methods-far-infrared ray, vacuum, and freeze drying and then enzymatically hydrolyzed using nine proteases: Alcalase, Flavourzyme, Kojizyme, Neutrase, Protamex, trypsin, α-chymotrypsin, and papain. In addition, the potential ability of hydrolysates to inhibit lipid accumulation in 3T3-L1 adipocytes was evaluated. The yield of hydrolysates from red sea cucumbers dried using each method was higher than that of the distilled water extract, and protein contents were either similar or higher. The hydrolysates that exhibited inhibitory effects on lipid accumulation, as demonstrated via Oil red O staining, were those obtained by far-infrared ray drying coupled with Alcalase, Flavourzyme, Kojizyme, or Neutrase treatment. In addition to the advantages of far-infrared drying and the characteristics of Flavourzyme, the Flavourzyme hydrolysate of far-infrared-dried red sea cucumber showed the highest inhibitory effect on lipid accumulation. In addition, this hydrolysate significantly decreased the expression of the protein factor fatty acid-binding protein 4, which is related to the late differentiation of 3T3-L1 adipocytes. Taken together, these results suggest that Flavourzyme hydrolysates from farinfrared-dried red sea cucumber may be used as a functional food and/or a pharmaceutical ingredient for the inhibition of lipid accumulation.

• Fisheries and Aquatic Sciences
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• v.19 no.5
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• pp.25.1-25.8
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• 2016

The fractionation of serine protease inhibitor (SPI) from fish roe extracts was carried out using polyethylene glycol-4000 (PEG4000) precipitation. The protease inhibitory activity of extracts and PEG fractions from Alaska pollock (AP), bastard halibut (BH), skipjack tuna (ST), and yellowfin tuna (YT) roes were determined against target proteases. All of the roe extracts showed inhibitory activity toward bromelain (BR), chymotrypsin (CH), trypsin (TR), papain-EDTA (PED), and alcalase (AL) as target proteases. PEG fractions, which have positive inhibitory activity and high recovery (%), were the PEG1 fraction (0-5 %, w/v) against cysteine proteases (BR and PA) and the PEG4 fraction (20-40 %, w/v) against serine proteases (CH and TR). The strongest specific inhibitory activity toward CH and TR of PEG4 fractions was AP (9278 and 1170 U/mg) followed by ST (6687 and 2064 U/mg), YT (3951 and 1536 U/mg), and BH (538 and 98 U/mg). The inhibitory activity of serine protease in extracts and PEG fractions from fish roe was stronger than that of cysteine protease toward common casein substrate. Therefore, SPI is mainly distributed in fish roe and PEG fractionation effectively isolated the SPI from fish roes.

• Journal of Microbiology and Biotechnology
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• v.19 no.11
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• pp.1393-1400
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• 2009

A bacteriocin-producing strain PI80 was isolated from the gut of Penaeus indicus (Indian white shrimp) and identified as Streptococcus phocae PI80. The bacteriocin was purified from a culture supernatant to homogeneity as confirmed by Tricine SDS-PAGE. Reverse-phase HPLC analysis revealed a single active fraction eluted at 12.94 min, and MALDI-TOF mass spectrometry analysis showed the molecular mass to be 9.244 kDa. This molecular mass does not correspond to previously described streptococcal bacteriocins. The purified bacteriocin was named phocaecin PI80 from its producer strain, as this is the first report of bacteriocin production by Streptococcus phocae. The bacteriocin exhibited a broad spectrum of activity and inhibited important pathogens: Listeria monocytogenes, Vibrio parahaemolyticus, and V. fischeri. The antibacterial substance was also sensitive to proteolytic enzymes: trypsin, protease, pepsin, and chymotrypsin, yet insensitive to catalase, peroxidase, and diastase, confirming that the inhibition was due to a proteinaceous molecule (i.e., the bacteriocin), and not due to hydrogen peroxide or diacetyl. Phocaecin PI80 moderately tolerated heat treatment (up to $70^{\circ}C$ for 10 min) and resisted certain solvents (acetone, ethanol, and butanol). A massive leakage of $K^+$ ions from E. coli $DH5\alpha$, L. monocytogenes, and V. parahaemolyticus was induced by phocaecin PI80, as measured by Inductively Coupled Plasma Optical Emission Spectrometry (ICPOES). Therefore, the results of this study show that phocaecin PI80 may be a useful tool for inhibiting L. monocytogenes in seafood products that do not usually undergo adequate heat treatment, whereas the cells of Streptococcus phocae PI80 could be used to control vibriosis in shrimp farming.

• Journal of Microbiology and Biotechnology
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• v.9 no.3
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• pp.282-291
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• 1999

Lactic acid bacteria were isolated from Kimchi and screened for bacteriocin. A total of 99 strains showed antimicrobial activity when grown on solid media, yet only 10 showed antimicrobial activity in liquid media. Strain H-559, identified as Lactococcus lactis subsp. lactis, exhibited the strongest inhibitory activity and was active against pathogenic bacteria including Listeria monocytogenes, Staphylococcus aureus, and Bacillus cereus as well as other lactic acid bacteria. The antimicrobial substance produced by L. lactis subsp. lactis H-559 was confirmed to be a bacteriocin by the treatment of $\alpha$-chymotrypsin, and protease type Ⅸ and ⅩIV. The bacteriocin activity remained stable between pH 2.0 and pH 11.0 and during heating for 10 min at $100^{\circ}C$. The bacteriocin production started in the exponential phase and stopped in the stationary phase. L. lactis subsp. lactis H-559 showed the highest bacteriocin activity at a culture temperature of $25^{\circ}C$, and an inverse relationship between the bacteriocin productivity and mean growth rate at different culture temperatures was observed. The mean growth rate and bacteriocin productivity of L. lactis subsp. lactis H-559 increased as the initial pH of the media increased.

• The Korean Journal of Physiology and Pharmacology
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• v.4 no.2
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• pp.129-135
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• 2000

Previously, we have reported that ${\rho}-chlorophenylalanine$ (PCPA), a serotonin depletor, profoundly increased pancreatic fluid and bicarbonate secretion but remarkably inhibited pancreatic amylase secretion in anesthetized rats. The present study was performed to verify the detailed effects of PCPA on pancreatic amylase synthesis that is directly related to amylase exocrine secretion. PCPA significantly decreased pancreatic RNA and protein contents as well as the amylase activity. However, pancreatic DNA content, trypsin and chymotrypsin activities were not influenced by the treatment of PCPA. The rate of pancreatic amylase synthesis, which was assessed by the amount of incorporated $[^{35}S]-methionine$ into amylase for 1 h, was also significantly decreased by 44% in PCPA-treated rats. In order to determine whether the PCPA-induced decrease of amylase synthesis resulted from change in the level of amylase mRNA, Northern blot analysis was performed. The mRNA expression level of amylase was also decreased by 48% in the PCPA-treated rats, indicating that the inhibitory effect of PCPA on the synthesis of pancreatic amylase was mainly regulated at a step prior to translation. It was also revealed in SDS-polyacrylamide gel electrophoresis that the qualitative change of amylase was induced by PCPA. The 54 KDa amylase band seems to be degraded into small molecular weight protein bands in PCPA-treated rats, suggesting that the PCPA- induced decrease of amylase may be partly attributed to the degradation of synthesized amylase.

• Korean Journal of Animal Reproduction
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• v.17 no.3
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• pp.183-191
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• 1993

These experiments were carried out to investigate whether the enzyme is involved in zona hardening during normal activatin of the oocytes by sperm, and demonstrate peroxidase activity during in vitro fertilization of oocytes treated with peroxidase inhibitors(250 $\mu$M phenylhydrazine, 28mM sodium sulfite, 350mM glycine ethyl ester and 50mM sodium azide) and tyrosine analogue(12.5mM tyramine). Also, zona soluble properties of the ovarian oocytes incubated for 0, 5, 10 and 15 hr in the presence of pheylhydrazine or tyramine were studied by using $\alpha$-chymotrypsin. The results obtained from these experiments were summarized as follows ; 1. The rates of fertilizatin in control oocytes and oocytes treated with phenylhydrazine or tyramine were 69.8%, 62.3% and 88.2%, respectively. However in vitro fertilization in oocytes treated with three different peroxidase inhibitors, sodium sulfite, glycine ethyl ester and sodium azide, were not induced. The oocytes treated with phenylhydrazine had no significant effect on in vitro fertilization rate as compared to control. However there was a significantly different in fertilization between tyramine treated group and control group(P<0.01). 2. The zona solubility(t50) of control and fertilized oocytes in culture treated with phenylhydrazine or tyramine were 30.7, 26.0 and 16.3 min., respectively. Phenylhydrazine treated group and tyramine treated group had effect on inhibition of zona hardening as compared to control group. These results suggest that ovoperoxidase is involved in zona hardening during normal activation of the oocytes by sperm. 3. t50 of control oocytes and ovarian oocytes treated with phenylhydrazine or tyramine for 5, 10 and 15 hr in vitro were 14.0, 26.2 and 32.0 min., 14.5, 26.9 and 30.2 min., and 14.0, 24.3 and 31.2 min., respectively. These results suggest that zona hardening in ovarian oocytes matured for various times in vitro cannot be inhibited by peroxidase inhibitors and tyrosine analogue, that the spontaneous zona hardening incultured ovarian oocytes is not caused by the secretory products of cortical granules released during the cortical reaction, ovoperoxidase.