• Korean Journal of Food Science and Technology
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• v.23 no.1
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• pp.62-67
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• 1991

The characteristics of Tongjinbyeo(Japonica) and Samgangbyeo($J{\times}Indica$) rice starches including physicochemical properties, differential scanning calorimetry and enzymatic digestion of lintnerized starches were investigated. Degree of hydrolysis of Tongjinbyeo starch with 2.2N HCI for 48 hr was higher than that of Samgangbyeo starch. Apparent first order reaction of starches was slow reaction for first period, and fast reaction second period on the datum point of acid treatment 24 hr. Absorbance at ${\lambda}_{max}$ and 680 nm, and ${\lambda}_{max}$ of iodine stained starch and amylose content decreased upon acid treatment. But water binding capacity, swelling power and solubility considerably increased as hydrolysis progressed. Relative crystallinity of two starches increased with acid treatment, and that of Tongjinbyeo starch was higher than that of Samgangbyeo starch. Differential scanning colorimetry(DSC) data continuously decreased for lintnerization periods, and those of Tongjinbyeo starch have higher than those of Samgangbyeo starch. The onset temperature of starch by DSC continuously decreased by treatment, but conclusion temperature increased until 24 hr and then decreased. The enthalpy for gelatinization decreased for both starches. Degree of hydrolysis of lintnerized Tongjinbyeo starch with glucoamylase was slightly higher than that of Samgangbyeo starch.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.365-368
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• 2003

Hair follicles develop as a result of epithelial-mesenchymal interactions between epidermal keratinocytes and dermal cells. Moreover hair follicles constitute multiple cells that influence hair follicle development and cyclic activity. We isolated some cells using explantation and enzymatic digestion method from human scalp hair follicles. So we could culture some follicular cells, such as outer root sheath (ORS) cells, dermal papilla (DP) cells, dermal sheath (DS) cells, matrix cells and melanocyte.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.356-358
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• 2003

The use of cultured epidermal sheet grafts for large full-thickness wound has been tempered by weak points such as long culture periods, difficulty in handling the fragile sheets, high costs and the detachment of the skin cells from the culture dishes by enzymatic digestion. To overcome the drawback of epidermal sheet grafts, we have developed a transplantation method to spray the cultured human keratinocytes with the mixtures of rhEGF and fibrin gel matrix to the full-thickness wounds on the dorsum of the athymic mice to regenerate epithelial layers. Wound biopsies were retrieved at 7, 14 and 21 days after transplantation and retrieved biopsies were analyzed by histology and immunohistochemistry. Transplanted keratinocytes and EGF-fibriin gel accelerated wound regeneration compared with control groups. The technique developed in this study overcomes the drawbacks of the current cultured epidermal sheet grafts and accelerates epidermal wound healing.

• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.92-94
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• 2003

The standard cultured sheet grafts for large full-thickness burn wound has been tempered by weak points such as long culture periods, difficulty in handling the fragile sheets, high costs and the detachment of the skin cells from the culture dishes by enzymatic digestion. Here we report on a new technique of using skin cells cultured on spherical microcarriers for skin regeneration. We cultured the human skin cells on biodegradable polymer microcarriers and transplanted onto wounds in the back of nude mice. After 21 days, histological examination showed the regeneration of epithelium. The technique developed in this study overcomes the drawbacks of the current artificial skin grafts, and could be utilized as an efficient skin wound regeneration therapy.

• Korean Journal of Food Science and Technology
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• v.6 no.2
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• pp.70-74
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• 1974

The round muscle of Korean cattle and squid muscle were cooked with various methods which were followed digestibility test by use of pepsin in-vitro, determination of amino nitrogen in the course of digestion procedure by using Formol method (AOAC) and influence of ether treatment for preminary test also examined. The results obtained were summarized as follows: 1. The order of digestibility values were demonstrated as follows: In case of beef, it was autoclaving, frying, raw, freezing, roasting, boiling and in case of squid muscle, it was raw, autoclaving, boiling, freezing, dry heating and roasting. 2. The amounts of amino nitrogen for beef and squid muscle were increased in proportion to digestibility value. 3. There were no significances in the digestibility between treating with ether and none of any treatment of beef and squid muscle in raw condition.

• Journal of Pharmacopuncture
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• v.16 no.1
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• pp.43-49
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• 2013

Objective: Pyungwi-san (PWS) plays a role in a number of physiologic and pharmacologic functions in many organs. Interstitial cells of Cajal (ICCs) are pacemaker cells that generate slow waves in the gastrointestinal (GI) tract. We aimed to investigate the beneficial effects of PWS in mouse small-intestinal ICCs. Methods: Enzymatic digestion was used to dissociate ICCs from the small intestine of a mouse. The whole-cell patch-clamp configuration was used to record membrane potentials from the cultured ICCs. Results: ICCs generated pacemaker potentials in the GI tract. PWS produced membrane depolarization in the current clamp mode. Pretreatment with a $Ca^{2+}$-free solution and a thapsigargin, a $Ca^{2+}$-ATPase, inhibitor in the endoplasmic reticulum, eliminated the generation of pacemaker potentials. However, only when the thapsigargin was applied in a bath solution, the membrane depolarization was not produced by PWS. Furthermore, the membrane depolarizations due to PWS were inhibited not by U-73122, an active phospholipase C inhibitor, but by chelerythrine and calphostin C, protein kinase C inhibitors. Conclusions: These results suggest that PWS might affect GI motility by modulating the pacemaker activity in the ICCs.

• BMB Reports
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• v.38 no.2
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• pp.225-231
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• 2005

Acid-labile subunit (ALS) is a component of the 150-kDa insulin-like growth factor-binding protein-3 (IGFBP-3) complex, which, by sequestering the majority of IGFs-I and -II and thereby prolonging the half-life of them in plasma, serves as a circulating reservoir of IGFs in mammalian species. A pGEX-2T plasmid and a baculovirus expression constructs harboring a coding sequence for glutathione-S transferase (GST)-porcine ALS (pALS) fusion protein were expressed in BL21(DE3) E. coli and Sf9 insect cells, respectively. The expressed protein was purified by glutathione or Ni-NTN affinity chromatography, followed by cleavage of the fusion protein using Factor Xa. In addition, pALS and hIGFBP-3 were also produced in small amounts in the Xenopus oocyte expression system which does not require any purification procedure. A 65-kDa pALS polypeptide was obtained following the prokaryotic expression and the enzymatic digestion, but biochemical characterization of this polypeptide was precluded because of an extremely low expression efficiency. The baculovirus-as well as Xenopus-expressed pALS exhibited the expected molecular mass of 85 kDa which was reduced into 75 and 65 kDa following deglycosylation of Asn-linked carbohydrates by Endo-F glycosidase, indicating that the expressed pALS was properly glycosylated. Moreover, irrespective of the source of pALS, the recombinant pALS and hIGFBP-3 formed a 130-kDa binary complex which could be immunoprecipitated by anti-hIGFBP-3 antibodies. Collectively, results indicate that an authentic pALS protein can be produced by the current expression systems.

• The Korean Journal of Physiology and Pharmacology
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• v.10 no.3
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• pp.131-135
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• 2006

Coeruleo-vestibular pathway which connects locus coeruleus and vestibular nuclei is noradrenergic. This study was designed to elucidate the effects of phenylephrine on the spontaneous activity of acutely isolated medial vestibular nuclear neurons of rat by whole-cell patch-clamp technique. Sprague-Dawley rats, aged 14 to 16 days, were used. After enzymatic digestion, dissociated medial vestibular neurons were transferred to a recording chamber mounted on an inverted microscope, and spontaneous action potentials were recorded by standard patch-clamp techniques. In current-clamp mode, the frequency of spontaneous action potential of medial vestibular nuclear neurons was decreased by phenylephrine (n=15). Phenylephrine increased the amplitude of afterhyperpolarization without changes in the resting membrane potential and spike width. In voltage-clamp mode, the whole potassium currents of the medial vestibular nuclear neurons were increased by phenylephrine (n=12). These experimental results suggest that ${\alpha}-receptor$ mediates the inhibitory effects on the neuronal activity of the medial vestibular nuclear neuron.

• Biomedical Science Letters
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• v.14 no.3
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• pp.179-186
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• 2008

Matrix metalloproteinases (MMPs), also designated matrixins, hydrolyze components of the extracellular matrix. These proteinases playa central role in many biological processes, such as embryogenesis, normal tissue remodeling, wound healing, and angiogenesis, and in diseases such as atheroma, arthritis, cancer, and tissue ulceration. In previous data, disruption of the actin cytoskeleton by cytochalasin D (CD) inhibited NO-induced apoptosis, dedifferentiation, cyclooxygenase (COX)-2 expression, and prostaglandin $E_2$ production in chondrocytes cultured on plastic or during cartilage explants culture. In this study, we investigated the effects of the actin cytoskeleton architecture on MMP-2 expression and dedifferentiation by CD in rabbit articular chondrocytes. Rabbit articular chondrocytes were prepared from cartilage slices of 2-weeks-old New Zealand white rabbits by enzymatic digestion. CD was used as a disruptor of actin cytoskeleton. In this experiments measuring CD dose response, primary chondrocytes were treated with various concentrations of CD for 24h. The actin disruption was determined by immunostaining. MMP-2 expression levels were determined by immunoblot analysis and Reverse transcriptase-Polymerase chain reaction (RT-PCR) and MMP-2 activity was determined by gelatin zymography. We found that cell morphological change and up-regulation of MMP-2 expression by CD as determined via immunostaining, gelatin zymography and immunoblotting. Moreover, CD induced MMP-2 transcription was detected by RT-PCR. Also, CD-induced type II collagen expression was inhibited by MMP-2 inhibitor I treatment. Our results indicate that CD up-regulated MMP-2 activation causes dedifferentiation of articular chondrocyte.

• The Plant Pathology Journal
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• v.35 no.6
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• pp.575-584
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• 2019

Colletotrichum acutatum is a species complex responsible for anthracnose disease in a wide range of host plants. Strain C. acutatum KC05, which was previously isolated from an infected pepper in Gangwon Province of South Korea, was reidentified as C. scovillei using combined sequence analyses of multiple genes. As a prerequisite for understanding the pathogenic development of the pepper anthracnose pathogen, we optimized the transformation system of C. scovillei KC05. Protoplast generation from young hyphae of KC05 was optimal in an enzymatic digestion using a combined treatment of 2% lysing enzyme and 0.8% driselase in 1 M NH₄Cl for 3 h incubation. Prolonged incubation for more than 3 h decreased protoplast yields. Protoplast growth of KC05 was completely inhibited for 4 days on regeneration media containing 200 ㎍/ml hygromycin B, indicating the viability of this antibiotic as a selection marker. To evaluate transformation efficiency, we tested polyethylene glycol-mediated protoplast transformation of KC05 using 19 different loci found throughout 10 (of 27) scaffolds, covering approximately 84.1% of the entire genome. PCR screening showed that the average transformation efficiency was about 17.1% per 100 colonies. Southern blot analyses revealed that at least one transformant per locus had single copy integration of PCR-screened positive transformants. Our results provide valuable information for a functional genomics approach to the pepper anthracnose pathogen C. scovillei.