• The korean journal of orthodontics
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• v.24 no.4 s.47
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• pp.933-943
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• 1994

The present study was undertaken to determine the effect of tensile force on DNA and protein biosynthesis in bone cells, and to identify the cell type(s) which primarily respond to external physical force among the heterogenous bone cell populations. As a prerequisite for this study, two bone cell populations which retain fibroblastic and osteoblastic feature were isolated from fetal rat calvaria with sequential enzyme digestion scheme. Tensile force was delivered to each bone cell population by two acrylic resin plates connected with a orthodontic expansion screw during culture period. Rate of DNA and protein synthesis in each bone cell population were assessed by the incorporated radioactivity of $[^3H]-thymidine$ into DNA and $[^3H]-proline$ into fraction of collagenase-digestible protein and noncollagenous protein, respectively. DNA synthesis of osteoblast-like calvarial cell populations was increased significantly by the application of tensile force for 24 hours. In contrast, no alteration in DNA synthesis of fibroblast-like populations could be observed in response to applied force. Tensile force induced the change in protein synthesis of bone cell populations with the same pattern. Total protein and collagen synthesis were increased whithin 24 hours in osteoblast-like populations, but not in fibroblast-like populations by tensile force application. These findings indicate that physical force can affect cellullar activity of the particular cell population, not all cell Populations residing in bone and osteoblasts respond more sensitively than fibroblasts. So osteoblasts can modulate the behavior of other bone cells including osteoclasts by producing several local regulating factors of bone metabolism. In this context, preferential responsiveness of osteoblasts to applied tensile force observed in this study suggests that osteoblasts may play an important role in regulation of physical force-induced remodelling process.

• Korean Journal of Food Science and Technology
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• v.25 no.4
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• pp.360-365
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• 1993

This study was carried out to examine the effects of phytate addition on the solubility and digestibility of the low-phytate soy protein isolate (LSPI) and high-phytate soy protein isolate (HSPI). In SDS-polyacrylamide gel electrophoresis of soy protein isolate, different patterns of proteins were observed in both HSPI and LSPI at various phytate and pH levels, suggesting that phytate may bind specifically to certain protein fractions at a particular pH. For example, proteins of M.W $1.8{\sim}3.5\;kDa$ resisted phytate binding at acidic pH. LSPI was fractionated into albumin, globulin, gliadin and glutelin, and phytate was shown to bind with difficultly to all three gliadin bands. Effects of phytate on the pepsin digestibility of soy proteins were apparent, especially in the short term digestion.

• Journal of Periodontal and Implant Science
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• v.38 no.sup2
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• pp.343-354
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• 2008

Purpose: Aggregatibacter actinomycetemcomitans is associated with localized aggressive periodontitis. It produces cytolethal distending toxin (CDT), which induces cell cycle arrest in the G2/M phase. The CDT holotoxin is composed of CdtA, CdtB, and CdtC. CdtB has structural homology to human DNase I and is an active component of the CDT complex acting as a DNase. In particular, the pattern homology seen in the CdtB subunit has been associated with specific DNase I residues involved in enzyme catalysis, DNA binding, and metal ion binding. So, to study the functions and regulation of recombinant CdtB, we made up a quantity of functional recombinant CdtB and tested it in relation to the metal ion effect. Materials and Methods: We constructed the pET28a-cdtB plasmid from A. actinomycetemcomitans Y4 by genomic DNA PCR and expressed it in the BL21 (DE3) Escherichia coli system. We obtained the functional recombinant CdtB by the refolding system using the dialysis method and then analyzed the DNase activity and investigated the metal ion effect from plasmid digestion. Results: The recombinant CdtB subunit was expressed as the inclusion bodies. We were able to obtain functional recombinant CdtB subunit using refolding system. We confirmed that our refolded recombinant CdtB had DNase activity and was influenced by the metal ions $Mg^{2+}$ and $Ca^{2+}$. Conclusion: We suggest that the factors influencing recombinant CdtB may contribute to CDT associated diseases, such as periodontitis, endocarditic, meningitis, and osteomyelitis.

• Proceedings of the Botanical Society of Korea Conference
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• 1985.08b
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• pp.51-57
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• 1985

In order to evlauate feasibility of the gene tagging by the maize transposable element Ac in heterologous plant systems, we have investigated physical distances and directions of transposition of the element in Arabidopsis thaliana and tobacco cultured cell line BY-2. We prepared a T-DNA construct that carried a non-autonomous derivative of Ac with a site for cleavage by endonuclease I-Scel (designated dAc-I-RS element). Another cleavage site was also introduced into the T-DNA region outside dAc-I-RS. A number of transgenic Arabidopsis plants were generated, each of which had a single copy of the T-DNA at a different chromosomal location. To examine the pattern of transposition, three out of these transgenic plants were crossed with the Arabidopsis plant that carried the gene for Ac transposase and progeny in which dAc-I-RS had been transposed were isolated. After digestion of the genomic DNA of these progeny with I-SceI, sizes of segment of DNA were determined byd pulse-field gel electrophoresis. We also performed linkage analysis for the transposed elements and sites of mutations near the elements. Our results with three transgenic lines showed that 50% of all transposition events had occurred within 1, 700 kilo-base pairs (kb) on the same chromosome, with 35% within 200 kb, and that the elements transposed in both directions on the chromosome with roughly equal probability. The data thus indicate that the Ac-Ds system is most useful for tagging of genes that are present within 200 kb of the chromosomal site of Ac in Arabidopsis. In addition, determination of the precise localization of the transposed dAc-I-RS element should definitely assist in map-based cloning of genes around insertion sites. In the present paper, we report typical examples of such gene isolation studies.

• BMB Reports
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• v.44 no.6
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• pp.387-392
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• 2011

To investigate the molecular scavenging capabilities of the larvae of Hermetia illucens, two serine proteases (SPs) were cloned and characterized. Multiple sequence alignments and phylogenetic tree analysis of the deduced amino acid sequences of Hi-SP1 and Hi-SP2 were suggested that Hi-SP1 may be a chymotrypsin- and Hi-SP2 may be a trypsin-like protease. Hi-SP1 and Hi-SP2 3-D homology models revealed that a catalytic triad, three disulfide bonds, and a substrate-binding pocket were highly conserved, as would be expected of a SP. E. coli expressed Hi-SP1 and Hi-SP2 showed chymotrypsin or trypsin activities, respectively. Hi-SP2 mRNAs were consistently expressed during larval development. In contrast, the expression of Hi-SP1 mRNA fluctuated between feeding and molting stages and disappeared at the pupal stages. These expression pattern differences suggest that Hi-SP1 may be a larval specific chymotrypsin-like protease involved with food digestion, while Hi-SP2 may be a trypsin-like protease with diverse functions at different stages.

• Applied Microscopy
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• v.28 no.4
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• pp.435-446
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• 1998

It is suggested that Urechis unicinctus is a filler feeder as like many tide and watery invertebrates which filtrate food materials by ciliary movement. However, the structure of the filter is not yet known in U. unicinctus, nor the filtering mechanism is not well understood. This study reveals ciliated epithelial cells in the foregut and the features of the cilia are good accord with that of known filtrating apparatus of other tide animals. This may implies that the foregut is in function of filtration and the food materials are filtrated by the ciliary movement. With the observation of the filtrating apparatus in the foregut, the intestine of U. unicinctus can be functionally compartmented into 3 parts. These are already known midgut and hindgut in function of digestion and respiration respectively, and the foregut in function of filtrating apparatus for foods. The filtrating apparatus of U. unicinctus is composed of the pseudostratified columnar epithelial cells with numerous cilia. The cilia are well differentiated kinocilia with the typical microtubule pattern, kinetosome and cilia roots. There are two kinds of striated cilia roots, the main root and the accessory root. The main root is extended perpendicularly from the cell surface to basement membrane and the short accessory root is branched with an acute angle of about $80^{\circ}$ from the main root at level of basal plate of the kinetosome. The spacial approaches of the main root with the large fused form of mitochondria is one of the characteristic features which might be in structural consideration an intimate association between energy source and energy mass consuming cell organelles.

• Korean Journal of Microbiology
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• v.24 no.1
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• pp.7-11
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• 1986

We screened lysates of the laboratory strains of pseudomonads utilizing hydrocarbon by agarose gel electrophoresis and cesium chloride-ethidium bromide equilibrium centrifugation, to find an intrinsic plasmid as a vector and to examine the relationship between the plasmid and hydrocarbon degradation. Only one strain from the examined strains, Pseudomonas putida KU190, contained a plasmid. We named the plasmid pKU41. The molecular size of pKU41 was determined as 41kb, using covalently closed circular forms of RP4 and pSY343 as standard size markers. The restriction sites of pKU41 for BamHI, BglII, EcoRI, HindIII, and SalI were 3, 1, 3, 6 and more than 13, respectively. With double or triple digestion, restriction map of pKU41 was constructed for BamHI, BglII and HindIII. For elucidation on the biological function of the plasmid, test was conducted on the ability of hydrocarbon utilization of the host strain but no apparent relationship was observed.

• ALGAE
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• v.21 no.2
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• pp.261-266
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• 2006

The innate soluble polysaccharides and phenolic compounds of marine macroalgae are serious contaminants which interfere with experimental procedures such as restriction enzyme digestion, polymerase chain reaction (PCR) and other enzymatic reactions using extracted DNA samples. The viscous polysaccharides are co-precipitated with DNA samples by isopropanol or ethanol precipitation in conventional experiment. To overcome the problem, a method for the isolation of high molecular weight DNA from Porphyra tenera is developed with the application of diatomaceous earth column. The isolated DNAs by this method were about 50-100 kb in size and could be digested well with restriction enzymes. The nuclease activity seemed to be minimal, and high reproducibility in the arbitrary primed PCR for RAPD analyses was a distinctive feature. These results suggest that this method is very efficient in isolating nucleic acid from macroalgae including Porphyra.

• Korean Journal of Food Science and Technology
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• v.28 no.1
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• pp.99-104
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• 1996

Carbohydrate-free caseinomacropeptide (CMP) was isolated from the sweet whey powder by a precipitation method using 12% trichloroacetic acid. The yield of carbohydrate-free CMP was 2.7 g from 100 g sweet whey powder. The electrophoretic pattern and the amino acid analysis of CMP showed that isolated CMP was quite pure, indicating the precipitation with 12% trichloroacetic acid was very effective for isolating carbohydrate-free CMP from the sweet whey powder. Trypsin, covalently immobilized on pore glass beads by carbodiimide (EDC) method, was 20mg per 1g glass beads. CMP was almost completely hydrolyzed by soluble trypsin in 24hr, but not by immobilized trypsin. The tryptic hydrolysates were fractionated on a Bio-Gel P 4 column $(1.5{\times}120\;cm)$and separated peptides were tested for their capacities to inhibit platelet aggregation using a aggregometer. The hydrolysate obtained from CMP after 24hr digestion by immobilized trypsin showed the highest activity.

• Journal of Nutrition and Health
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• v.15 no.4
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• pp.301-312
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• 1982

A series of growing and digestion trials was conducted using Sprague- Dawley weanling male rats in order to determine the effects of two protein sources (casein and ISP (isolated soyprotein) ), three protein levels (10, 20 and 30%), and two feeding patterns (ad libitum and meal feeding) on the growth, protein and energy metabolism, and body composition of rats. The total energy level of experimental diets was kept constant in 3,600 kcal ME /kg diet. The results were as follows : 1) The amount of food intake and the weight gain of meal-fed group were lower than those of ad libitum group. Though the intake of meal-fed group on 20 and 30% casein diet was only 85% of ad libitum group, it was able to gain as much as ad libitum group. 2) There were no significant differences in the food efficiency ratio (FER) and the energy efficiency (weight gain per 100 kcal GE intake) between ad libitum and meal feeding group. The FER and the energy efficiency of 20 and 30% casein diets of meal-fed group were greater than those of ad libitum group. 3) Though the gross energy intake (GE ), the digestible energy (DE) and the metabolizable energy (ME) tended to be lower at meal-fed group, the DE/GE and the ME/GE ratios for meal-fed group were the same as those for ad libitum. 4) Though meal- fed group fed less amount of nitrogen than ad libitum group, there were no differences in nitrogen balance and the retention of rats among the treatments. Actually meal-fed group retained more nitrogen than ad libitum group at the levels of 20 and 30% dietary protein. 5) After growing and digestion trials, the body composition of rats was constant among all treatments. Significantly high negative correlation coefficient (r = -0.77) was found between the body fat content and the body moisture content. Consequently, this study suggests that meal- fed group on 20 and 30% casein diets has shown more effective utilization of the ingested food and energy than ad libitum group, and increasing tendency of weight gain and the body fat deposition. Those influences of meal feeding pattern in rats were more effective on the casein diet than on the ISP diet.