• Korean Journal of Fisheries and Aquatic Sciences
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• v.12 no.4
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• pp.225-234
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• 1979

For the effective utilization of diverse and abundant resource of seaweeds in Korea as a food protein supplment, extraction conditions of water, salt, and alkali soluble proteins were investigated in previous work(Ryu and Lee, 1977: Lee et al., 1977: Lee et al., 1978). The present study as a part of the serial work was thus aimed to find the conditions of isolation and purification of extracted proteins, and to evaluate the nutritional quality of the isolated seaweed proteins in terms of amino acid composition, chemical score, protein score, modified essential amino acid index(MEAAI), and in vitro digestibility presented as pepsin-pancreatin digest residue index (PPDRI). As for the isolation of extracted proteins, TCA treatment was more effective for the proteins from rhodophyceae and Chlorophyceae while the precipitation at isoelectric point was more desirable for Phaeophyceae proteins. In amino acid composition, water soluble protein fraction was superior to the other fractions in Porphyra suborbiculata whereas both water and alkali soluble fractions seemed to bo more benefitial for Enteromorpha linza and Ulva pertusa; the extraction with alcohol-alkali mixed solvent for Undaria pinnatifida and Sargassum fulvellum. Glutamic acid and aspartic acid content was particularly high in all protein fractions examined. The total amino acid content of Porphyra suborbiculata and Enteromorpha linza was almost equivalent to that of dried whole egg although the essential amino acid content was lower. A comparative analysis was made on the inedexes between raw seaweed powder and isolated protein. Chemical score of Porphyra suborbiculata and Ulva pertusa was approximately 35 and 56 in cafes of raw powder and isolated protein respectively while only 10 to 16 for raw powder of Undaria pinnatifida and Sargassum fulvellum and 30 to 35 for their isolated proteins. Protein score of all isolated proteins was in the range of 63 to 73 which indicates that isolated protein would be mere valuable than the fern of raw seaweed powder. Digestibility by means of PPDRI was found to be extremely low in case of raw powder but it could be doubled in case of isolated protein yielding 67 to 70 for Porphyra suborbiculata and Ulva pertusa.

• Korean Journal of Veterinary Research
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• v.37 no.2
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• pp.321-330
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• 1997

Enrofloxacin is one of the second-generation quinolones which have been widely used to treat bacterial infections in various species including chicken, pig, horse and cattle. The objective of the present study was to describe the serum bactericidal activity(SBA) of enrofloxacin, its pharmacokinetic behaviors after intramuscular or intravenous administration to Korean native goats in the dose rate of 5mg/kg b.w. The results obtained through this study were as follows : 1. Sera collected from both sexes of Korean native goats administered 5mg/kg i.v. or i.m. showed potent antibacterial activities up to the 12 hours by way of the serum bactericidal activity. 2. Concentrations of enrofloxacin in the biological samples were measured by high-performance liquid chromatography(HPLC) so as to study pharmacokinetic characteristics. For detection of enrofloxacin, 10% TCA was optimal for protein precipitation and the mobile phase was 0.01M citric acid/methanol/acetonitrile(7/2/1, pH 3.5) with solid phase being the $C_{18}$ reversephase column and detection wavelength being 278nm. The limit of detection of enrofloxacin on HPLC was $0.05{\mu}g/ml$. 3. Pharmacokinetic profile of enrofloxacin administered 5mg/kg i.v. in Korean native goats was best described by two-compartment open model and that administered i.m. the same rate by one-compartment model. There were no sex differences in pharmacokineticl parameters. In conclusion, enrofloxacin showed potent in vivo antibacterial activity and excellent pharmacokinetic properties in Korean native goats, hence it may be used as a potential antibacterial in the veterinary clinical settings.

• Applied Biological Chemistry
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• v.23 no.1
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• pp.14-22
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• 1980

White and black sesame produced in Korea were defatted with ethyl ether or n-hexane. Defatted sesame meal was extracted with water and salt solution, and protein extraction was precipitated at various pH 1 through 12, with trichloro acetic acid (TCA), tannic acid and ammonium sulfate, respectively. Protein was purified by Sephadex A-25, G-75, G-100 and G-200, and identified its protein fraction by polyacrylamide gel electrophoresis. Amino acids composition of protein in white sesame was analyzed by automatic amino acid analyzer. Protein contents of white sesame, black sesame and sesame meal are 20.5%, 19.2%, and 44.7%, respectively. n-Hexane was the most suitable solvent for extraction of oil from sesame. Crude protein precipitation was better in higher pH. The protein extraction was more effective with the solution containing sodium chloride tinder the pH 8. Globulin in total protein was high and prolamin was less than in other cereal proteins. Glutamic acid contents of white sesame and sesame globulin were 17.1%, and 20%, respectively. Both proteins contained relatively high levels of essential amino acids. 12-13 bands were found in water soluble protein and 2 bands in salt soluble protein were detected by the disc gel electrophoresis, and were identified in both of white and black sesame. The salt soluble protein of white sesame could be purified by Sephadee G-100 and G-200.

• The Korean Journal of Nuclear Medicine
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• v.28 no.1
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• pp.106-111
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• 1994

2-iminothiolane is known to bind $NH_2$ group of lysine in the protein and deliver SH group, which can be used to label protein with $^{99m}Tc$. In this study, we looked for the best reaction condition in which 2-iminothiolane is conjugated to human polyclonal IgG and labeling condition with $^{99m}Tc$-glucoheptonate. Labeling yield was measured with TSK G4000SW column and HPLC or precipitation with 10% TCA (trichloroacetic acid) and 1% HSA. In vivo distribution was investigated with Staphylococcal abscess bearing rats. With decreasing glucoheptonate, the labeling yield decreased. Without 2-iminothiolane, $^{99m}Tc$-glucoheptonate was bound to IgG, which seemed to be direct labeling. With increasing 2-iminothiolane upto 20 times higher than IgG, the labeling yield increased, and plateau was seen with higher molar excess of 2-iminothiolane. Polymer formation was not observed. The pH for the conjugation of 2-iminothiolane and IgG was best around 6.4. $^{99m}Tc$-2-iminothiolane-IgG showed faster blood clearance, higher renal activity and lower hepatic and splenic activity than $^{99m}Tc$-DTPA-IgG. The biodistribution of $^{99m}Tc$-2-iminothiolane-IgG with higher molar excess of 2-iminothiolane was not different from that with lower molar excess. Labeling antibodies with $^{99m}Tc$ using 2-iminothiolane can afford a possible route to simple labeling and wide clinical use of the immunoscintigraphy.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.9 no.3
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• pp.155-162
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• 1976

Forty one samples from thirty three species of algae (19 from 15 species of Rhodophyceae, 18 from 14 species of Phaeophyceae, 3 from 3 species of Chlorophyceae, and 1 of marine Phanerogams) collected from several locations on the east, west and south coast of Korea, were analyzed for their contents of crude protein, fat, cellulose, ash, nitrogen free extract, amino nitrogen, and total amino acids. For the examination of extractability of algal protein with water, 4 species of algae, Sargassum thunberggi, Grateloupia filicina, Phyllospadix japonica, and Sargassum confusum, were analyzed. And the effect of some precipitation treatments for isolation of algal protein was also tested. As a matter of fact, Rhodophyceae showed high content in crude protein and low in crude fat while the case was opposite for Phaeophyceae and Chlorophyceae. Refering to the content of crude protein and total amino acids, the recommendable algae for protein sources were Sargassum thunbergii, Acrosorium flabellata, Phacelocarpus japonicus, Laurencia okamurai, Laurencia intermedia, Grateloupia filicina, Chondrus ocellatus, Gloiopeltis furcata, Gigartina tenella, Dictyota dichotoma, and Scytosiphon lomentaria. Methanol treatment appeared most effective in precipitation isolation of protein from water extracts whereas pH control method did not so beneficial. The precipitation rate of protein was particularity higher in the extract of Sargassum confusum ana the lowest was makted from the extract of Sargassum thunbergii.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.10 no.3
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• pp.151-162
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• 1977

Distribution of marine algae is diverse in Korea and the resource of edible algae is abundant marking 239,037 tons of yearly production in 1976. They have been known as a protein source and used as a supplement in Korean diet. It is necessary to estimate the potentiality and properties of usable algal proteins especially as food resources and studies of extraction and separation of the proteins, therefore, are basically required for this purpose. In this study, the influence of various factors including the sample treatment, extraction time and temperature, sample us extraction solvent ratio and pH upon the extractability of the water soluble protein was determined. And the effect of precipitation treatment for isolation of the algal protein from the extracts was also tested. Nine species of algae, the major ones in consumption as food namely Porphyra suborbiculata, Undaria pinnatifida, Hizikia fusiforme, Sargassum fulvellu, Enteromorpha linza, Codium fragile, Sargassum kjellmanianum and Ulva pertusa were collected as fresh from Kijang, Yangsan Gun, in the vicinity of Busan city. The content of crude protein $(N\times6.25)$ of the algae ranged from $9.46\%\;to\;24.14\% showing the highest value in Porphyra suborbiculata and the minimum in Hizikia fusiforme. In the effort of maceration of blending methods on the extractability, immersion freezing in dry ice-methanol solution appeared most effective yielding 1.5 to 2.5 times extractability than that of the mortar grinding method. The effect of the ratio of sample vs solvent on extractability differed from species. It was enhanced at the ratio of 1:20 (w/v) in Ulva pertusa and Enteromorpha linza while the ratio was 1:30 (w/v) for Cedium fragile, Undaria pinnatifida, Hizikia fusiferme, Sargassum fulvellum and Porphyra suborbiculata and 1:40 for Sargassum kjellmanianum respectively. The effect of extraction time and temperature was revealed differently from species which might be caused by differences in the constitution of algal tissues resulting in that the extraction for 1 hour at $50^{\circ}C$ gave the maximum extractabilily in Ulva pertusa and Enteromorpha linza, 2 hours in Porphyra suborbiculata, Hikikia fusiforme, Undaria pinnatifida, Sargassum kjellmanianum and 3 hours in Codium fragile. And the extractability was higher at $50^{\circ}C$ to $60^{\circ}C$ for the most of the tested samples except Hizikia fusiforme. The optimum pH for the extraction was 9 to 12. The recovery of extractable nitrogen to the total nitrogen was $63\%$ in average with the first extracts and $8.6\%$ with the second extracts respectively. Both extracts were prepared by 2 hour extraction at $50{\pm}1^{\circ}C$ with dry ice-methanol frozen and seasand macerated materials. And these conditions assumed to be an optimum for the extraction of water soluble algal proteins since the nitrogen content after the first extraction covered $90\%$ of the total water extractable nitrogen. In the precipitation of the extracted proteins, Barnstein method and methanol treatment seemed to be more efficient than other precipitation methods.

• Food Science of Animal Resources
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• v.29 no.5
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• pp.633-639
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• 2009

The principal objective of the current study was to prepare low molecular weight peptides from milk proteins using enzymatic hydrolysis techniques, in an effort to assess the antioxidant activity of these peptides. The casein and whey proteins isolated from fresh milk were treated with several proteolytic enzymes, such as chymotrypsin, pepsin, and trypsin and the resulting low molecular weight peptides were collected by TCA precipitation. Their identity was confirmed by SDS-PAGE analysis. The hydrolysis experiments indicated that whey protein treated with chymotrypsin displayed the highest degree of protein hydrolysis. The antioxidant activity of milk protein hydrolysates was determined by measuring the ABTS-radical scavenging activity. The results of these experiments showed that hydrolysis of the milk protein was effective in increasing their antioxidant activities. Especially, the tryptic digested casein displayed the highest radical scavenging activity (80.7%). The hydrolyzed low molecular weight milk protein was isolated using an ultrafiltration membrane. The casein hydrolysate passed through a membrane with molecular weight cut-off (MWCO) of 3 kDa displayed the strongest antioxidant activity.

• Journal of Periodontal and Implant Science
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• v.27 no.2
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• pp.425-441
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• 1997

The neuropeptide substance P(SP) has been implicated in the mediation of inflammation and immune-mediated disease such as arthritis. Recently, it was reported that SP was markedly increased around the blood vessels in inflamed gingiva as well as in close association with the inflammatory cell infiltrate. These results support that SP may contribute to the pathophysiology of neuronal inflammation in human periodontal tissues. SP may regulate inflammatory/immune responses by stimulating the proliferation of human T cells, differentiation and antibody-secreting potential of B cells, macrophage respiratory burst, connective tissue proliferation, and the secretion of cytokines from monocytes and T cells. Here, I studied potential role of SP as a costimulatory chemical signal in inflammatory/immune responses, by determining the released proinflammatory cytokines such as $MIP-1{\alpha}$, $IL-1{\beta}$, and IL-6 from culture supernatants of homogeneous immune cell lines. Serum free cell supernatants were concentrated with TCA precipitation, fractionated with SDS-PAGE, and subjected into western blot analysis. Among 15 cell lines tested, macrophage/monocyte cell line RAW264.7 and WRl9m.1 showed the highest level of induction of $MIP-1{\alpha}$ when stimulated with LPS. Discrete IL-6 bands with multiple forms of molecular mass were detected from supernatants of B cell lines A20(32kDa), Daudi(32, 35kDa), and SKW6.4(29kDa), which were expressed constitutively. $IL-1{\beta}$ could not be detected by the method of western blot analysis from supernatants of all cell lines tested except RAW264.7, WRl9m.1, and erythroid cell line K562 which showed the least amount of $IL-{\beta}$ secretion. SP $10^{-9}M$ with suboptimal dose of LPS treatment showed synergistic induction of $MIP-1{\alpha}$ release from RAW264.7 or WR19m.1, and also IL-6 release from A20, but this synergism is not the case in costimulation of RAW264.7 or WRl9m.1 with SP $10^{-9}M$ and TPA. Although treatment of T cell line CTLL-R8 with SP $10^{-7}M$ or PHA+TPA induced modest level of $MIP-1{\alpha}$ secretion, synergism was not observed when they are applied together. These findings all together suggest the possibility of a regulatory role of SP in inflammatory/immune reaction through differential modulation of bioactivities of other chemical cosignals.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1995.04a
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• pp.89-89
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• 1995

Surface epithelial cells isolated from hamster tracheas and grown on a thick collagen gel become a highly enriched population of mucus-secreting cells. Epithelial cells from tracheas of hamsters were collected using enzymatic procedures and cultured under various conditions. The medium used consisted of a 1:1 mixture of medium 199 and Dulbecco's modified Eagle's (DME) medium which was conditioned before use. Insulin, transferrin, hydrocortisone, epidermal growth factor, and extract from bovine hypothalamus were used as supplement. Due to relatively low basal rates of min secretion from in vitro cultures, cultures are generally radiolabeled using $^3$H-glucosamine as a metabolic precursor. The radiolabeled mucinsreleased are quantitated by precipitation with TCA/PTA. Using this cell culture system, we investigated mucin release of goblet cells by altering the media bathing the apical surface of hamster tracheal surface epithelial(HTSE) cells. Acidic media added sulfuric acid caused sigcificant increases in mucin relesse (155${\pm}$20% at pH 4 and 146${\pm}$16% at, pH 5). Ammonium hydroxide also increased mucin release at pH 9.0(156${\pm}$17%) and pH 10(295${\pm}$9%) respectively. This additional mucin release seems to be associated with cell membrane damage as indicated by release of cellular LDH. SP stimulates secretion of mucin in cultured HTSE cells(154${\pm}$16% at 1${\times}$10$\^$-6/M and 165${\pm}$25% at 1${\times}$10$\^$-5/M. PAF at 5${\times}$10$\^$-6/M and 5${\times}$10$\^$-5/M enhanced by HTSE cells in vitro 168${\pm}$34% and 259${\pm}$30% of mucin secretion, respectively. The increase in mucin release by PAF and SP was not secondary to cell damage or necrosis. SP and PAF may be in mediating mucous secretion induced by inflammation irritantion and infection.