• Parasites, Hosts and Diseases
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• v.37 no.1
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• pp.39-46
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• 1999

As a preliminary study for the explanation of pathobiology of Neodiplostomum seoulense infection. a 54 kDa protease was purified from the crude extract of adult worms by sequential chromatographic methods. The crude extract was subjected to DEAE-Sepharose Fast Flow column, and protein was eluted using 25 mM Tris-HC1 (pH 7.4) containing 0.05. 0.1, 0.2 and 0.4 M NaC1 in stepwise elution. The 0.2 M NaCl fraction was further purified by Q-Sepharose chromatography and protein was eluted using 20 mM sodium acetate (pH 6.4) containing 0.05, 0.1. 0.2 and 0.3 M NaCl, respectively. The 0.1M NaCl fraction showed a single protein band on SDS-PAGE carried out on a 7.5-15% gradient gel. The proteolytic activities of the purified enzyme were specifically inhibited by L-trans-epoxy-succinylleucylamide (4-guanidino) butane (E-64) and iodoacetic acid. The enzyme, cysteine protease. showed the maximum proteolytic activity at pH 6.0 in 0.1 M buffer, and degraded extracellular matrix proteins such as collagen and fibronectin with different activities. It is suggested that the cysteine protease may playa role in the nutrient uptake of N. seoulense from the host intestine.

• The Korean Journal of Food And Nutrition
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• v.33 no.3
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• pp.223-236
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• 2020

In this study, a safety evaluation was conducted to confirm if the Enterococcus faecium CKDB003 strain obtained by selection from a mixed fermentation of fruit and milk is suitable for use as a probiotic. The MIC value for the 10 antibiotics specified in the EFSA guidance was below the acceptable cut-off value. The antibiotic resistance genes aac(6')-li, eatAv, and msr(C) exist by whole genome sequencing, but are in the chromosome and not in the plasmid, thus confirming that there is no possibility of transmission to other microorganisms. It was confirmed that cytolysin (cylA, cylB, cylI, cylL-l, cylL-s, cylM, cylR1, cylR2), aggregation substance (asa1, asp1), collagen adhesion (ace), enterococcal surface protein (esp), endocarditis antigen (efaA), hyaluronidase (hyl) and gelatinase (gelE) were not present in the genome by examining the genes of factors related to virulence. Also, the biochemical analysis showed no toxic enzyme activities, and no virulence genes were detected by the PCR method. Thus, the E. faecium CKDB003 strain can be safely used as a health functional food probiotic, based on the results of the safety assessment.

• Journal of Plant Biotechnology
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• v.37 no.1
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• pp.96-101
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• 2010

Leaf explants of the Solanum hainanense plant, grown in vitro, were cultured in basal Murashige and Skoog (MS) media supplemented with 0.5 mg/L kinetin and 1 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) for callus initiation. For maintenance and proliferation, the callus was cultured on MS medium supplemented with 1 mg/L 6-benzylaminopurine (BAP) and 0.5 mg/L 2,4-D. The glycoalkaloid content in the callus was at its maximum after ten weeks of culture (188.65 mg/g), whereas that of the one-year-old control was 22.22 mg/g in the root and 5.99 mg/g in the stem. The glycoalkaloid extracted from the callus inhibited the activity of collagenase on collagen gel. High performance liquid chromatography (HPLC) analysis showed that biotransformation occurred when a callus was grown on medium supplemented with various carbon sources. These results suggest that callus of S. hainanense is a good material for production of glycoalkaloid.

• Journal of Microbiology and Biotechnology
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• v.15 no.2
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• pp.425-427
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• 2005

The steam distillate obtained from Thujopsis dolabrata var. hondai sawdust was fractionated by centrifugal thin-film evaporation, and the fractions were then investigated for antiplatelet activity using washed rabbit platelets. The biologically active constituent of T. dolabrata var. hondai sawdust was isolated by silica gel column and HPLC chromatographies and characterized as carvacrol by various spectral analyses. Carvacrol inhibited platelet aggregation induced by collagen, arachidonic acid, and platelet activating factor with IC$_{50}$ values of 12.6, 2.5, and 385.3 $\mu$M, respectively. However, carvacrol had no effect on thrombin, calcium ionophore A23l87, or phorbol l2-myristate l3-acetate induced platelet aggregation. Carvacrol was a much more potent inhibitor, as antiplatelet agents, compared with aspirin. These results suggest that carvacrol isolated from T. dolabrata var. hondai sawdust may be useful as a lead compound for inhibiting arachidonic acid-induced platelet aggregation.

• The Korean Journal of Pharmacology
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• v.26 no.2
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• pp.193-207
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• 1990

Transforming growth factor ${\beta}(TGF-{\beta})$ is a multifunctional polypeptide with diverse effects on the proliferation, differentiation and other functions in many cell types. $TGF-{\beta}$ is highly abundant in bone matrix and induces divergent responses in many aspects of bone cell metabolism . Several lines of investigation indicate that matrix-associated $TGF-{\beta}$ is the products of bone cells themselves. However, exact bone cell type reponsible for the production of $TGF-{\beta}$ is still in controversy, The present study was undertaken to determine the cellular origin of matrix-associated $TGF-{\beta}$ and to assess how different bone cells respond to $TGF-{\beta}$. As a prerequisite for this, 5 bone cell populations of distinct phenotype were isolated from fetal calvaria with sequential enzyme digestion protocol and biochemical characterization. Calvarial cell populations released in early stage showed fibroblastic features whereas populations relesed later was enriched with osteoblast-like cell as judged by their acid and alkaline phosphatase activities, cAMP responsiveness to parathyroid hormone, calcitonin and prostaglandin $E_2$ and collagen synthesis rate. By polyacylamide gel and immunoblot analysis of bone and calvarial cell extracts, presence of $TGF-{\beta}$ in bone tissues and production of $TGF-{\beta}$ by bone cells were confirmed again. Subsequent analysis of calvarial cell extracts prepared as individual population revealed that all calvarial cell populations synthesize $TGF-{\beta}$. Exogenously added $TGF-{\beta}$ induced biphasic response upon bone cell proliferation under serum-free condition. In osteoblastic cell populations, it was stimulatory whereas inhibitory in fibroblastic cell populations. In contrast, collagen and noncollagen protein synthesis of all calvarial cell populations were stimulated by $TGF-{\beta}$. Enhancement of protein synthesis was found to be more general rather than specific for collagen synthesis. In addition, effects of $TGF-{\beta}$ on protein synthesis were independent to its effects on cell proliferation. In summary, production of $TGF-{\beta}$ by bone cells and differential actions on various cell populations observed in this study suggest that $TGF-{\beta}$ may play an important role in the regulation of bone metabolism by modulating the specific cellular functions in autocrine and paracrine fashion.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.43 no.6
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• pp.598-605
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• 2010

This study investigated the physicochemical and enzymatic properties of unmarketable bastard halibut (Paralichthys olivaceus) cultured in different regions (i.e., Jeju, Wando, and Geoje) as a potential source of surimi and surimi gel. The proximate composition of unmarketable bastard halibut cultured in different regions did not differ significantly at P<0.05. Compared to Alaska pollock muscle, all of the unmarketable bastard halibut muscle had a 4% higher crude protein content and 5% lower moisture content. The collagen content of bastard halibut muscle cultured in Jeju was 1.96 g/100 g, which was higher than in fish cultured in other regions. Regardless of the region where cultured or pH, the enzymatic activities of the crude extracts from unmarketable bastard halibut muscle ranged from 0.30.0.48 U/mg for casein and hemoglobin, 11.9.13.7 U/mg for LeuPNA, 5.6.6.7 U/mg for ArgPNA, 2.8.4.7 U/mg for SAAPFNA, and 0.1.0.2 U/mg for BAPNA. Regardless of region, no mercury or lead was found in any of the unmarketable bastard halibut muscle, except for lead in fish cultured in Geoje. The strength of surimi gels from unmarketable bastard halibut cultured in Jeju, Geoje, and Wando was 1059, 988, and 900 g${\times}$cm, respectively. The surimi gel from unmarketable bastard halibut cultured in Jeju was stronger than commercial Alaska pollock surimi, which was grade SA.

• Journal of Veterinary Clinics
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• v.32 no.3
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• pp.224-230
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• 2015

This study was designed in the fabricated poly (L-Lactic-co-${\varepsilon}$-Caprolactone) (PLCL) scaffold using chitosan-alginate hydrogel, which would be more suitable to maintain the biological and physiological functions continuing three dimensional spatial organizations for chondrocytes. As a scaffold, hydrogels alone is weak at endure complex loading within the body. In this study, we made cell hybrid scaffold constructs with poly (L-Lactic-co-${\varepsilon}$-Caprolactone) (PLCL) scaffold and hydrogels to make a three-dimensional composition of cells and extracellular matrix, which would be a mimic of a native cartilage. Using a particle leaching technique with NaCl, we fabricated a highly-elastic scaffold from PLCL with 85% porosity and $300-500{\mu}m$ pore size. A mixture of bovine chondrocytes and chitosan-alginate gel was seeded and compared with alginate as a control on the PLCL scaffold. The cell maturation, proliferation, extracellular matrix synthesis, glycosaminoglycans (sGAG) production and collagen type-II expressions were better in chondrocytes seeded in chitosan-alginate hydrogel than in alginate only. These results indicate that chondrocytes with chitosan-alginate gel on PLCL scaffolds provide an appropriate biomimetic environment for cell proliferation and matrix synthesis, which could successfully be used for cartilage repair and regeneration.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.19 no.3
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• pp.265-286
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• 1997

Bone morphogenetic proteins(BMPs) are a group of transforming growth factor beta(TGF-${\beta}$)-related factors and multifunctional proteins, especially the only known biologic factors capable of inducing endochondral bone formation at an extraskeletal site. This study was performed to investigate the effect of the partially purified porcine BMP(pBMP) at an ectopic site. PBMP was partially purified from porcine bone matrix and its activity was monitored by an in vivo bioassay. The purification method utilized extraction of the bone-inducing activity with 4M guanidine, followed by chromatography on heparin-Sepharose. Active fractions were assayed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. And the fractions were reconstituted with inactive insoluble collagenous bone matrix from rats, acid soluble type I collagen from rat tail and chondroitin-6-sulfate sodium salt and implanted into the pectroralis muscle pouches of Sprague-Dawley rats. And the carrier complex was implanted on the opposite side as control. The rats were sacrificed at the day of 1st, 3rd, 5th, 7th, 11th, 14th and 21st after implantation and examined histologically, radiologically and biochemically. And alkaline phosphatase activity and calcium content were used as indices of bone formation. The results were as follows ; 1. Active fractions were localized in a zone between 31 and 40 KDa on SDS-PAGE. 2. The implanted 3.0mg of the partially purified pBMP induced cartilage and bone in the muscle tissue of rats through an endochondral ossification process. 3. Inactive insoluble bone matrix, type I collagen and chondroitin-6-sulfate have functioned as carriers for pBMP, but revealed some foreign body reactions. 4. Soft X-ray didn't reveal significant change between the experimental and the control group. 5. The alkaline phosphatase activities in the experimental group of 5th, 7th, 11th, 14th and 21st were increased significantly compared with control (p<0.01) with the peak in the group of 11th day. 6. With time, the calcium content of the experimental group increased. And the calcium contents in the experimental group of 11th, 14th and 21st were increased significantly compared with control (p<0.01).

• Applied Biological Chemistry
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• v.47 no.1
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• pp.130-134
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• 2004

The arils of Euphoria longana L. was extracted with 80% aqueous MeOH, and the concentrated extract was partitioned with EtOAc, n-BuOH and $H_2O$. From the n-BuOH fraction, four compounds were isolated through the repeated silica gel, ODS and Sephadex LH-20 column chromatographies. From the result of physico-chemical data including NMR, MS and IR, the chemical structures of the compounds were determined as 1,1-dimethyl-2propenyl $1-O-{\beta}-D-glucopyranoside$, ethyl ,${\beta}-D-glucopyranoside$, 5-(hydroxymethyl)-2-furfur-aldehyde and uridine. Uridine exhibited inhibition effect of 79% on platelet aggregation at the concentration of$5\;{\mu}g/ml$.

• Journal of Conservation Science
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• v.34 no.3
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• pp.157-166
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• 2018

In this study, gelatin binding ability was increased by adding cross linking agent to improve adhesive characteristic of animal glue. Animal glue added genipin measured gel strength and viscosity, the structural analysis, the color retention degree, elution degree, and rupture strength. And the water resistance and ultraviolet light resistance with the addition of genipin were compared. As a result of the study, the gel strength and viscosity increased with the amount of genipin. As a result of the structural analysis, in gelatin, the absorption peak of the triple structure of collagen structurally stabilized was observed. As a result of the color retention degree, the film was observed because of the lowered brightness. The amount of elution glue was increased with addition of genipin at $50^{\circ}C$ distilled water condition and rupture strength has increased with the amount of genipin. In the water resistance and light fastness, there was no appearance before and after deterioration due to the addition of genipin. Based on the results of this study, it confirmed the adhesive characteristics of animal glue added genipin and examined the experimental method applicable for animal glue. After the addition of genipin, flexibility, re-solving, adhesive force, and curing speed, which are unique characteristics of glue, can be improved without disappearing, so it is expected that it will be applicable to production of animal glue and conservation of cultural heritage when homogeneous glue is secured.