• Food Science of Animal Resources
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• v.36 no.6
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• pp.769-778
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• 2016

In this study, we improved the eggshell-membrane separation process by separating the shell and membrane with EDTA solution, evaluating effects of three different extraction solutions (acetic acid, EDTA, and phosphate solution), and co-purifying multiple eggshell proteins with two successive ion-exchange chromatography procedures (CM Sepharose Fast Flow and DEAE Sepharose Fast Flow). The recovery and residual rates of eggshell and membrane separated by the modified method with added EDTA solution were 93.88%, 91.15% and 1.01%, 2.87%, respectively. Ovocleidin-116 (OC-116) and ovocalyxin-36 (OCX-36) were obtained by loading 50 mM Na-Hepes, pH 7.5, 2 mM DTT and 350 mM NaCl buffer onto the DEAE-FF column at a flow rate of 1 mL/min, ovocleidin-17 (OC-17) was obtained by loading 100 mM NaCl, 50 mM Tris, pH 8.0 on the CM-FF column at a flow rate of 0.5 mL/min. The purities of OCX-36, OC-17 and OC-116 were 96.82%, 80.15% and 73.22%, and the recovery rates were 55.27%, 53.38% and 36.34%, respectively. Antibacterial activity test suggested that phosphate solution extract exhibited significantly higher activity against the tested bacterial strains than the acetic acid or EDTA extract, probably due to more types of proteins in the extract. These results demonstrate that this separation method is feasible and efficient.

• Food Science of Animal Resources
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• v.34 no.1
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• pp.26-32
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• 2014

This study was conducted to examine the effects of eggshell membrane hydrolysates (ESMH) on the anti-inflammatory, anti-wrinkle, anti-microbial activity, and moisture-protection for cosmetic use. Whole ESMH (before fractionation), and fraction I (>10 kDa), fraction II (3-10 kDa), and fraction III (<3 kDa) of the hydrolysates were assessed in this experiment. As lipopolysaccharide (LPS) and IFN-${\gamma}$ caused the inflammation on Raw264.7 cell, whole ESMH and fraction I showed to be effective in inhibiting the induction of cell inflammation depending on the concentration, and also showed outstanding effect to suppress the skin inflammation. Fraction I inhibited collagenase and elastase activities to a greater extent than the other fractions, while all fractions had antibiotic effects at concentrations of 10 mg/disc and 20 mg/disc. In addition, it showed the moisture protection effects of skin on the holding amount and losing amount of moisture in upper-inner arm of the human body with a relatively low loss rate in skin, which confirmed that the hydrolyzed fractions of ESM helps to form the superior protective layer of moisture. It was concluded that ESMH fractions with different molecular weights, especially the 10 kDa fraction, have anti-lipopolysaccharide, anti-IFN-${\gamma}$-induced inflammation, anti-collagenase and elastase activities, and thus can be used as a cosmetic agent to protect skin.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.9 no.1
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• pp.43-55
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• 1976

Fertilization and early development of turbo cornutus was studied based on the samples which were collected in Yeosu area. Particular emphasis was paid on induction of artificial spawing, fertilization rate, preembryonic development, the growth of the early larva and larval survival to various salinity. Among the various methods for induction of artificial spawning which have been tested for the present study, drying by exposure to air is the. most efficient, and percentage fertilization rate was $83.8-96.4\%$. The diameter of fertilized eggs was $0.182{\pm}0.0028mm$; and the diameter of egg membrane was $0.245{\pm}0.093mm$. Under the temperature range of $20.6-25.4^{\circ}C$ the larvae hatched out after 11:05-11:15 hours of fertilization. After 3.0-3.5 days of fertilization the planktonic larvae begand to settle, and the settlement terminated within 5 days. During the period of 150 days of early culturing the diameter growth of shell(M) and the diameter of shell aperture(A) was formulated as follows: $$1972\;M=0.33e^{0.02070D}$$ $$A=0.19e^{0.02282D}$$ $$1973\;M=0.32e^{0.02282D}$$ $$A=0.16e^{0.02596D}$$ During the same period of early culturing the relative growth of shell diameter and the diameter of shell aperture was formulated as follows : 1972 A=0.6478 S-0.1575 1973 A=0.5897 S-0.0515 After 11 days of larval hatching $0.02-0.18\%$ of planktonic larvae settled. After 150 days of settlement the survival rate of the early shells was $7.4-21.6\%$. Under the temperature range of $21.0-22.7^{\circ}C$ the optimum salinity range for the development of egg and the planktonic larvae was $30-35\%_{\circ}$.

• Journal of Korean Ophthalmic Optics Society
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• v.7 no.1
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• pp.39-43
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• 2002

This study was investigated the anti-angiogenic activities of Cnidium officinale Makino and Tabanus bovinus by using Chorioallantoic membrane (CAM) and rat cornea. First experiment, the fertilized chicken eggs were kept in a humidified egg incubator at $37^{\circ}C$. After 4 days incubation, about 3 ml of albumin was aspirated from eggs with an hypodermic needles through a small hole drilled at the narrow end of the eggs, and the shell membrane on the floor of the air sac was peeled away. Embryos with chorioallantois of 3~5 mm in diameter were employed for the assay of antiangiogenic activity. Retinoic acid was used as a positive control for this experiment. After 48 h of treatment. branching pattern of blood vessels below coverslip containing retinoic acid ($10{\mu}g/egg$) was dramatically decreased. A simiar angiogenic inhibition was observed in the CAM treated with $50{\mu}g/egg$ of Cnidium omcinale Makino and Tabanus bovinus extracts. Second experiment, rat corneal neovascularization was induced by suturing (one stitch) the cornea with 10-0 nylon, and terramycin was applied on the cornea for I week to prevent corneal inflammation. In the cornea of rats untreated with herbal extracts, numerous vessels were usually seen invading the cornea by day 2 or 3 after suture, and reaching the lesion area within 5~6 days. Corneal neovascularization was gradually increased and peaked at 3 weeks. In contrast to this, herbal extracts conspicuously inhibited the angiogenesis, Oral administration of herbal extracts (20 mg/kg body weight/day) for 4 weeks significantly inhibited the rat corneal angiogenesis induced by suture, and the length of blood vessels in herbal medicine-treated rat cornea was conspicuously lower than that in control animals. A similar phenomenon was also observed in the rat cornea treated with thalidomide (200 mg/kg body weight/day). These findings indicate the anti-angiogenic properties of Cnidium officinale Makino and Tabanus bovinus, suggesting that these properties may be one of the pharmacological mechanisms underlying the anti-tumor and anti-metastatic activities of herbal extracts tested in this study.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.3
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• pp.454-458
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• 2014

In this study, we examined the ionization rate and permeability of nanocalcium prepared from oyster shells with various particle sizes. Four particle sizes of the calcium samples were prepared by centrifugation according to their density disparity in alcoholic solution: NC (normal calcium), C-1 (supernatant of 1,000 rpm), C-2 (supernatant of 2,000 rpm), and C-3 (supernatant of 3,000 rpm). Particle sizes of NC, C-1, C-2, and C-3 were $2,280.3{\pm}64.3nm$, $521.3{\pm}83.3nm$, $313.9{\pm}29.5nm$, and $280.0{\pm}3.4nm$, respectively. C-3 showed a slight increase in ionization rate compared with the other calcium samples, but their differences were not significant. Dialysis membrane-employed analysis showed that nanocalcium permeability increased as its particle size smaller; 32% of C-3 nanocalcium was transported to the outside of the membrane, whereas C-1 showed a 25% transport rate. We determined the permeability of the nanocalciums by using rat intestinal sacs, in order to provide different intestinal environments depending on pH level. Nanocalcium generally showed a higher permeability at pH 7, which represents an ileum environments compared to the duodenum and jejunum environments at pH 4.2 and pH 6.2, respectively. However, C-3 calcium showed the highest permeability, followed by C-2, C-1 and NS calciums. This result shows that the size of calcium positively affected its permeability in the intestinal sac. Taken together, nano-sized calcium derived from discarded oyster shell shows improved permeability in intestinal environments.

• Current Research on Agriculture and Life Sciences
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• v.1
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• pp.67-83
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• 1983

The new microsporidia S80 isolated from, Bombyx mori L. in Korea showed ovoid in the morphology of the spores and the size were measured $2.9{\pm}0.28{\mu}$ in length and $1.7{\pm}0.29{\mu}$ width. No other microsporidian spore like this has not been so far isolated from Silkworm. The length of the polar filament extruded in hydrogen peroxide ($H_2O_2$) at $30^{\circ}C$ was $26{\mu}$ of a round cytoplasm on the top. The spores were partly stained with Giemsa, Safranin-O and Gram as the same staining properties as Nosema bombycis, Microsporidia K 79 and other microsporidian spores. The fine structures were observed under scanning eleceron microscope through ultrathin sectioning. The spore wall was composed of three layers ; the thin exospore of an electron dense rippled layer, the thick electron lucent endospore which was thinning considerably at the polar filament insertion point, and the inner limiting membrane. Polar cap present at the sporeapex, with a long polar filament of 12-13 coils, subtending angle of $60^{\circ}$ to spore axis, which is tubular made up of a multilayered and are a benes core, light ring structure enclosing the dance core, the dark ring structure enclosing the inner light ring structure and the other than and light ring structure bounded from cytoplasm. Lamellate polaroplast occupied the anterior part of the spore, and the two neclei with dense nucleoplasm bounded by a double nuclear envelope were cited in the slight downer middle portion of spore. From the characteristics of the shape, size and fine structures, it is certain to reason the Microsporidia S80 belong to the phylum Microspora, class Microspora, order Microsporida, order Microsporida. The shape of two nuclei cited seems to be genus Nosema, but in the classification for the suborder it should be defined wheather pansporoblasts be formed or not and for the genis especial attempts have been made to define the characters which distinguish the disporous genera in the life cycle. Survey through the infection of the bad cocoons during 1980 to 1982 in South Korea the areas contaminated with new microsporidia were revealed 5 provinces of Kyung-Gi, Kang-Won, Chung-Nam and Chun-Nam. Pathological effects inoculated per os at second instar larvae of silkworm, the LD 50 was $7.1{\times}10^7/ml$ as lower pathogenecity than that of Nosema bombycis Naegeli of $1.2{\times}10_7/ml$. While on the other hand the inoculation of the microsporidia at fourth instar larvae lowerd the whole cocoon weight and cocoon shell weight and significant at 1% level. The microsporidia S80 defined it can not be transmitted transovarially from the result of predictive and collective examination of 21 egg batches from the infected female moth.