• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.4
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• pp.646-652
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• 2004

With the aim of using a cosmetic material, chondroitin sulfates extracted from midduck tunics (Styela clava) and munggae tunics (Halocynthia roretzi) were examined in vitro with two cell lines for cell toxicity, collagen synthesis, cell growth and recovery ability after U.V. irradiation. Cell toxicity test with A 431 and CCD 1108Sk was able to observe high activity between 400 and 600 $\mu\textrm{g}$/m while standard chondroitin sulfate (CS) purchased from Sigma was showed at 80 $\mu\textrm{g}$/mL. Even fraction 1 and 2 collected from chondroitin sulfates originated from midduck appeared having the highest activity between 600 and 1000 $\mu\textrm{g}$/mL, but slightly lower compared to crude chondroitin sulfates from both mideduck and munggae. In cell growth examination, it was not able to find significant differences between chondroitin sulfates used. Both crude chondroitin sulfates were exhibited the highest activity for two cell lines except that of mideduck which was showed activity for CCD 1108Sk. CS, fraction 1 and 2 from midduck were not able to demonstrate a significant activity in collagen synthesis. On the contrary, crude chondroitin sulfates from both munggae and midduck were showed the highest activity at 100 and 50 $\mu\textrm{g}$/mL with only CCD 1108Sk. The recovery ability after U.V. irradiation with crude chondroitin sulfates from both munggae and midduck were showed high activity at 400 $\mu\textrm{g}$/mL with CCD 1108Sk and A 431. But there were no activity observed in fractions examined, As a consequence, the crude chondroitin sulfates from both munggae and midduck might not only be available as a cosmetic material but also useful for increasing some activity by blending properly.

• Korean Journal of Environmental Biology
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• v.32 no.3
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• pp.216-224
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• 2014

Decrease in dissolved oxygen concentrations associated with temperature fluctuation is an important criteria to evaluate the mortality rate of the species. Based on this parameter, we investigated the survival rate, physiological response and histological change of warty sea squirt. It was found that the survival rate of the warty sea squirt species was 63.3% at $23^{\circ}C$ and 56.6% at $26^{\circ}C$ respectively. However, exposure of six days at $29^{\circ}C$ caused deaths among species, which indicated the 6day-$LT^{50}$ of the tested species to be $24.58^{\circ}C$ ($19.48{\sim}35.48^{\circ}C$). Further, after 11 day of exposure, the dissolved oxygen concentration has been found to decrease, with the survival rate of 20% at $4.0mg\;L^{-1}$ and deaths at $2.0mg\;L^{-1}$, thus 11day-$LC^{50}$ calculated to be $3.88mg\;L^{-1}$ ($3.29{\sim}4.57mg\;L^{-1}$). In addition, decrease in rate of oxygen consumption and excretion of ammonia was also noted at this critical water temperature and dissolve doxygen concentration. Moreover, there has been common histopathological changes were observed in warty sea squirt's gill pouch, digestive tract, and tunic as follows such as: proliferation of epithelial cells, condensation and necrosis, permeation of phagocyte and blood cell, loss of cilium and muscular fiber degeneration. Based on our study results, we suggest that these parameters can also be useful to evaluate the survival rate and physiological response in other species.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.1
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• pp.71-76
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• 2002

To understand the effect of heated effluent from a nuclear power plant on marine organism, experimental culture of Halocynthia roretzi was carried out at heated effluent of Wolsung nuclear power plant from January to December 1996. Temperature was $11.2\~27.9^{\circ}C$ and salinity was $32.54\~34.59\%_{\circ}$ during the culture period, The Growth of H. roretzi on lower area of Bonggil-ri (St. 1) was not normal in height, breadth and weight due to heated effluent. Daily growth rate (DGR) of H. roretzi about Kampo area (St, 4) was significant other station, and St. 1 was significant from other station except St. 4, Mytilus edulis was major fouling organism (over $90\%$) that were M. edulis, Dideninum moseleyi, Styela clava in experiment culture farm. St. 1 was higher (mean 143 individual) and St. 4 was lower (mean 56 individual) appearance attached of M. edulis. Growth of H. roretzi reduced when attached number of M. eduiis was increased, because correlation between DGR and number of M. edulis was negative.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.4
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• pp.641-645
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• 2004

Crude chondroitin sulfates extracted from midduck tunics (Styela clava) and munggae tunics (Halocynthia roretzi) were examined in vivo in order to be utilized as a cosmetic material which was followed by an in vitro assay. Examinations, such as acute oral toxicity, skin sensitization, acute eye irritation, and primary skin irritation, were peformed with a variety of laboratory animals. Phototoxic and photosensitization tests were not conducted since all chondroitin sulfates failed to absorb U.V. light at the range of 280 to 420 nm. In acute dermal and eye irritation, both specific clinical signs and dead cases were not demonstrated during the test period, but crude chondroitin sulfates from midduck and munggae tunics, and standard chondroitin sulfate from bovine trachea were showed 2.5, 1 and 1.25 of acute ocular irritation index (A.O.I.), respectively. In the case of skin sensitization, crude chondroitin sulfate from midduck tunics exhibited neither specific clinical signs nor dead cases in the entire course of the examination. While in acute oral toxicity, crude chondroitin sulfates from both midduck and munggae tunics found neither specific clinical signs nor dead cases during the test, and LD50 was suspected to be over 2 g/kg. Based on this study, it was proven that crude chondroitin sulfates from either midduck or munggae tunics can be used safely as a cosmetic material.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.32 no.4
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• pp.295-307
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• 2006

Styela clava, called non-native tunicate or sea squirt, is habitat which include bays and harbors in Korea and several sites in the sea faced world. We fabricate cellulose membrane nerve conduit (CMNC) from this native sea squirt skin, and evaluate the capacity of promoting peripheral nerve regeneration in the rat sciatic nerve defect model. After processing the pure cellulose membrane from the sea squirt skin as we already published before, CMNC was designed as a non-tubular sheet with 14 mm length and 4 mm width. Total eleven male Spraque-Dawley rats (12 weeks, weighing 250 to 300g) were divided into sham group (n=2), silicone tube grafted control group (n=3) and experimental group (n=6). Each CMNC grafted nerve was evaluated after 4, 8 and 12 weeks in the experimental group, and after 12 weeks, sciatic function was evaluated with sciatic function index (SFI) and gait analysis, and histomorphology of nerve conduit and the innervated tissues of sciatic nerve were all examined using image analyzer and electromicroscopic methods in the all groups. The regenerated axon and nerve sheath were found only in the inner surface of the CMNC after 4 weeks and became more thicker after 8 and 12 weeks. In the TEM study, CMNC grafted group showed more abundant organized myelinated nerve fibers with thickened extracellular matrix than silicone conduit grafted group after 12 weeks. The sciatic function index (SFI) and ankle stance angle (ASA) in the functional evaluation were $-47.2{\pm}3.9$, $35.5^{\circ}{\pm}4.9^{\circ}$ in CMNC grafted group (n=2) and $-80.4{\pm}7.4$, $29.2^{\circ}{\pm}5.3^{\circ}$ in silicone conduit grafted group (n=3), respectively. And the myelinated axon was 41.59% in CMNC group and 9.51% in silicone conduit group to the sham group. The development of a bioactive CMNC to replace autogenous nerve grafts offers a potential and available approach to improved peripheral nerve regeneration. As we already published before, small peptide fragment derived from the basement membrane matrix proteins of squirt skin, which is a kind of anchoring protein composed of glycocalyx, induced the effective axonal regeneration with rapid growth of Schwann cells beneath the inner surface of CMNC. So the possibilities of clinical application as a peripheral nerve regeneration will be able to be suggested.

• Journal of the Korean Society of Food Science and Nutrition
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• v.29 no.5
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• pp.922-934
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• 2000

To investigate the composition of carotenoids present in marine organisms and the biological activity of the carotenoids, carotenoids of the muscles and tunic of tunicates and shellfishes were isolated and identified. Anitmutagenic activities of the carotenoids for S. typhimurium TA 98 and cytotoxic activity for cancer cell lines were determined. Total carotenoid contents in the muscle of tunicata ranged from 18.65 mg% to 2.39 mg%. The highest amount of the total carotenoid was found in the muscle of Halocynthia aurantium, followed by Styela clava (HERDMAN), H. roretzi, H. hilgendorfi f. igaboya, H. hilgendorfi f. retteri, S. plicata (LESUEUR) in order. Interestingly, total carotenoid content in the muscle of S. clava (HERDAMAN) was higher than that of H. roretzi. Total carotenoid content of all tunicata, other than H. aurantium and H. roretzi, were higher in muscle than tunic. The major carotenoids in H. roretzi, H. aurantium, S. plicata (LESUEUR), and S. clava (HERDAMAN) were cynthiaxanthin (25.1∼42.2%), halocynthiaxanthin (9.7∼26.3%), diatoxanthin (8.0∼18.7%) and β-carotene (7.7%∼21.7%). Similarly, cantaxanthin (19.6%), cynthiaxanthin (15.4%), halocynthiaxanthin (14.8%), and (3R, 3'R), (3S, 3'S)-astaxanthin (22.6%) in H. hilgendorfi f. retteri and fucoxanthin (26.6%), cynthiaxanthin (21.8%), halocynthiaxanthin (15.2%), and β-carotene (9.3%) in H. hilgendorfi f. igaboya were major carotenoids in both tunicate. However, the composition of carotenoids in muscle and tunic of tunicata was similar each other. Among the shellfishes examined, total carotenoid content of the muscle of Peronidia venulosa (Schrenck) and Corbicula fluminea, and of the gonad of Atrina pinnata and Chlamys farreri, was ranged from 2.51 to 6.83 mg% which were relatively higher than that of other shellfishes. The composition of the carotenoids of shellfishes, which might depend upon their living environments, was varied. But cynthiaxanthin (15.9∼39.0%) and zeaxanthin (9.6∼21.9%) in gonad of C. farreri, and muscles of Buccinum Volutharpa perryi (JAY) and Crassostrea gigas, cynthiaxanthin (21.5∼48.6%) and mytiloxanthin (14.6%) in muscle of C.fluminea and gonad of A. pinnata, and canthaxanthin (60.6%) and isozeaxanthin (20.5%) in muscles of P. venulosa (Schrenck), and β-carotene (23.7%∼37.8%) and zeaxanthin (18.2∼20.4) in muscles of Semisulcospira libertina and Meretrix lusoria were major carotenoids. Interestingly, diester type-carotenoids were present along with free type-carotenoids in muscles of C. gigas. antimutagenic effect of the carotenoids isolated from tunicata and shellfishes against 2-amino-3-methylimidazol [4,5-f]quinoline (IQ) for S. typhimurium TA 98 was proportional to the amount (20, 50 and 100㎍/plate) treated. Mutagenicity of IQ was significantly reduced by astaxanthin, isozeaxanthin, mytiloxanthin and halocynthiaxanthin, whereas the mutagenicity of aflatoxin B₁(AFB₁) was significantly reduced by β-carotene, isozeaxanthin, and mytiloxnthin. Growth inhibition effect of carotenoids isolated from tunicata and shellfishes for cancer cell was proportional to the amount (5, 10, and 20㎍/plate) treated. The growth of HeLa cell by β-carotene, cynthiaxanthin, astaxanthin and halocynthiaxanthin, NCI-H87 cell by β-carotene, astaxanthin, cynthiaxanthin, and halocynthiaxanthin, HT-29 cell by β-carotene, cynthiaxanthin, mytiloxanthin and halocynthiaxanthin, and MG-63 cells by β-carotene, cynthiaxanthin, astaxanthin, canthaxanthin and halocynthiaxanthin were statistically reduced.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.53 no.2
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• pp.244-250
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• 2020

Microplastic pollution in the marine ecosystem has been emerged as a global issue. In this study, we investigated the abundance of microplastics from clam Meretrix lusoria, blood arkshell Scapharca broughtonii, and warty sea squirt Styela clava obtained from a local market in Busan, Korea. The marine organisms were digested in 10% KOH, and were incubated at 40℃ and 150 rpm, for 7 days. The digest was filtered through standard sieve (5 mm, 1 mm, 300 ㎛, and 100 ㎛), and mciroplastics were identified using a light microscope and microFT-IR. The abundance of microplastics of clam, blood arkshell, warty sea squirt was 0.08 items/g, 0.05 items/g, and 0.12 items/g, respectively. The predominant microplastic size was in the range of 100-300 ㎛, occupying 48%, and the predominant type was fiber. The composition of microplastics was mostly rayon, semi-synthetic cellulosic material and polyester, which are main component of fabric and textile. We strongly believe that this preliminary work may provide useful information for the establishment of the standardized analysis method of microplastics ingested by marine organisms.

• Culinary science and hospitality research
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• v.13 no.3
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• pp.68-79
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• 2007

The purpose of this study was to contribute to merchandising of native local foods. This study was designed to standardize the recipes and to analyze the nutrients of native local foods in Gijang region such as 'Myeolchijjigae', 'Galchijjigae' and 'Maejipjjim'. The test recipe for each food was prepared according to the information obtained from the personal interview of Gijang natives and then supplied for the sensory evaluation. After that, CAN Pro 2.0 was used for the nutritional evaluation. The results are as follows : Myeolchijjigae was preferred when radishes and traditional soybean paste were added. Galchijjigae was more liked when young pumpkins, mixed soy sauce and soybean paste were added. Maejipjjim was preferred with Aeji(Codium adhaerens), Warty sea squirt(Styela clava), soybean paste and powdered perilla seed(Perilla frutescens var. japonica) added. The analysis of nutrient composition revealed that in general energy content was low whereas protein content was as high as recommended requirement in Myeolchijjigae and Maejipjjim. Calcium and phosphorus were high in Myeol-chijjigae, vitamin A in Galchijjigae, and vitamin A, B_6$, folic acid, phosphorus and iron in Maejipjjim.

• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.475-479
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• 2003

Glycosaminoglycans(GAGs) from sea squirt, Styela clava was extracted with sodium phosphate at $105^{\circ}C$ for 2 hr and deprotein with trichloroacetic acid or hydrochloride. This GAGs was mainly constituted of galactose, glucosamine, glucose, mannose and galactosamine, and was phenylalanine, threonine, glutamic acid and aspartic acid. Mineral contents was mainly constituted 3.0mg% sodium, 1.6mg% potassium and 1.2mg% phosphorus and heavy metal was not detected. At pharmaceutical and cosmetic code of GAGs, protein and sulfate contents should included each range $14.0{\sim}22.0%$, $35.0 {\sim}45.0%$. After 5.0% trichloroacetic acid(w/v) and 10.0% HCl(v/v) treatment, protein and sulfate contents of GAGs was contained each 35.1%, 35.4% and each 22.0%, 18.5%.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.41 no.6
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• pp.458-465
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• 2008

The nutritional contribution of mideodeok extracts (ME) on rockfish (Sebastes schlegelii) feed and fish muscle was investigated. Different concentrations of the ME mixed with commercial diet were fed to mature rockfish for 8 weeks. The lipid and ash contents of the formulated diets were relatively similar to the control diet, while increasing the extract concentration increased the moisture content and decreased the protein contents. Major fatty acid components (C18:1n-9, 16:0, C20:5n-3, C22:6n-3) were of comparable quantity. High presence of C18:2n-6 was attributed to soybean oil incorporated in the diets, while the essential fatty acids were within limits (0.9-1.0%). The diet fortified with 6% ME produced the highest feed efficiency, with increased protein content in the muscle as well as lipid content for both muscle and liver. Hepato- and visceral-somatic index values were elevated with increasing ME concentration Muscle fatty acid contents were mostly C18:1n-9 and C16:0, with low absorption of C18:2n-6 in both the muscle and liver. Total highly unsaturated fatty acid content was significantly reduced in the fish muscle, but the values were higher for fish fed with a ME-fortified diet. An increasing trend for eicosapentaenoic acid and docosahexaenoic acid was also observed with increased ME fortification, with liver levels of these compounds remaining within range throughout the duration of the experiment.