• Molecules and Cells
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• v.37 no.2
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• pp.178-186
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• 2014

Differential transcription of the clusterin (CLU) gene yields two CLU isoforms, a nuclear form (nCLU) and a secretory form (sCLU), which play crucial roles in prostate tumorigenesis. Pro-apoptotic nCLU and anti-apoptotic sCLU have opposite effects and are differentially expressed in normal and cancer cells; however, their regulatory mechanisms at the transcriptional level are not yet known. Here, we examined the transcriptional regulation of nCLU in response to hypoxia. We identified three putative hypoxia response elements (HREs) in the human CLU promoter between positions -806 and +51 bp. Using a luciferase reporter, electrophoretic gel mobility shift, and chromatin immunoprecipitation assays, we further showed that hypoxia-inducible factor-$1{\alpha}$ (HIF-$1{\alpha}$) bound directly to these sites and activated transcription. Exposure to the hypoxia-mimetic compound $CoCl_2$, incubation under 1% $O_2$ conditions, or overexpression of HIF-$1{\alpha}$ enhanced nCLU expression and induced apoptosis in human prostate cancer PC3M cells. However, LNCaP prostate cancer cells were resistant to hypoxia-induced cell death. Methylation-specific PCR analysis revealed that the CLU promoter in PC3M cells was not methylated; in contrast, the CLU promoter in LNCap cells was methylated. Co-treatment of LNCaP cells with $CoCl_2$ and a demethylating agent promoted apoptotic cell death through the induction of nCLU. We conclude that nCLU expression is regulated by direct binding of HIF-$1{\alpha}$ to HRE sites and is epigenetically controlled by methylation of its promoter region.

• Clinical and Experimental Pediatrics
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• v.45 no.9
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• pp.1106-1113
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• 2002

Purpose : Nuclear ($factor-{\kappa}BNF-{\kappa}B$) is now recognized as playing a potential role in programmed cell death and the adaptive response to various stress. Cellular hypoxia is a primary manifestation of many cardiovascular diseases. It seems that vascular endothelial growth factor (VEGF) and insulin like growth factor-I(IGF-I) have a function as a protective molecule in the heart against several stress including hypoxia. In this study, the role of $NF-{\kappa}B$ to the cellular response and regulation of protective molecules against the acute hypoxia in the heart was studied. Methods : To cause acute hypoxic stress to the heart, Sprague Dawley rats were exposed to hypoxic chamer($N_2$ 92% and $O_2$ 8%). After the hypoxic exposure, nuclear proteins, total proteins and mRNA were isolated from heart. Translocation of the transcription factors $NF-{\kappa}B$, NF-ATc, AP-1 and NKX-2.5 were evaluated by electrophoretic mobility shift assay(EMSA). The expression of IGF-I and VEGF were studied before and after the hypoxic stress by competitive-PCR, Northern hybridization and Western hybridization. To confirm the role of the $NF-{\kappa}B$ in the heart, the rats also were pretreated with diethyl-dithiocarbamic acid(DDTC) into peritoneal cavity to block $NF-{\kappa}B$ translocation into nucleus. Results : The expression of $NF-{\kappa}B$, AP-1 and NF-ATc were increased by the hypoxic stress. Increased expression of the VEGF and IGF-I were also observed by the hypoxic stress. However, the blocking of the $NF-{\kappa}B$ translocation reduced those expressions of VEGF and IGF-I. Conclusion : These results suggest that $NF-{\kappa}B$ has a protective role against the acute hypoxia through several gene expression, especially VEGF and IGF-I in heart muscle.

• Journal of Sericultural and Entomological Science
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• v.23 no.2
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• pp.11-36
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• 1982

Forty-nine plant species as additives to silkworm artificial diet and 5 species as cellulose sources for artificial diet were screened for their economic values as feed-resources for the silkworm. Feeding response to artificial diet was tested on 82 silkworm strains. The effect of rearing conditions on feeding response and enzyme activities in the silkworm was investigated. The results were summarized as follows. 1. Seven species out of 49, Vigna sinensis ENDL, Ipomoea vatatas Lamarck, Cyperus anuricus Var. Laxus, Alnus japonica Stendel, Trifolium repens L, Prunus serrulata Lindley. Var, Glycine max L increased feeding response, compared with the basic formula of artificial diet. 2. The economic values of Vigna sinensis ENDL, Ipomoea vatatas Lamarck, Cyperus anuricus Var. Laxus, Ainus japonica Stendel, Cassia tera L, Erigeron canedensis L as feed-resources for artificiale diet were recognized, through feeding experiment during the entire larval stage. 3. Mulberry cellulose showed the best results in rearing and cocoon characteristics. 4. The extent of feeding response varied according to strains and varieties. Varieties in japanese strains showed higher feeding response than those in chinese and european varieties, with considerable variations among a varieties in strains. 5. The begining of 4th instar seems to be a proper time to convert from mulberry to artificial diet, or artificial diet to mulberry, however the middle of 3rd instar seems acceptable. 6. The optimum temperature for artificial diet rearing is 30$^{\circ}C$ during the period of 1st-3rd instar and 28$^{\circ}C$ for 4th-5th instar. 7. Electrophoretic isozyme patterns of esterase and acid phosphatase on agarose gel, as affected by strain. rearing temperature and feed-resources, were observed as follow. (1) Isozyme patterns of mid-gut esterase varied, depending on instar. One or two more isozyme bands were observed in the larvae than feed on the mulberry fed for the artificial diet. (2) A strain, chinese-15 with a higher feeding response, had 1∼2 more bands than chinese-60 with a lower feeding response. (3) Five bands of mid-gut esterase in 3rd and 4th instar larvae reared at 28$^{\circ}C$. and 4 for 3rd instar and 6∼7 for 4th instar larvae at 35$^{\circ}C$ were observed. (4) No similar esterase bands could be found among mid-gut, blood and silkgland. There are five esterase bands in the midgut, one in blood and three in silkgland. (5) There was rather small digerence in acid phosphatase types of mid-gut and blood according to varieties and rearing temperature. No active band was shown in silkgland. In midgut, there was one acid phosphatase band at 3rd instar, two at 4th instar and three at 5th instar. In blood, one active band at 3rd or 4th instar and three bands at 5th inster wire detected.

• Journal of Korean Society of Forest Science
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• v.58 no.1
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• pp.1-7
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• 1982

Megagametophyte and embryo tissue of Pinus densiflora were subjected to study the inheritance of glutamate-oxalate transaminase(GOT) and leucine aminopeptidase(LAP), and linkage relationship among isozyme loci coding both enzymes by starch gel zone-electrophoresis. Four zones of activity were observed for GOT. No variation was found in the fastest migrating zone (GOT-A). Electrophoretic phenotypes of the other two zones (GOT-B and GOT-C) showed 1:1 segregation ration, suggesting that each zone is controlled by a single locus. Foru and three alleles were identified at both loci respectively. The isozyme pattern of the fourth zone(GOT-D), migrated cathodally, coincided precisely with that of GOT-C. Whether the two zones are controlled by the same locus or by two tightly linked loci remained unknown. In all three variant GOT zones, heterozygoes embryos produced triple band patterns, indicating that GOT isozyme in Pinus densiflora is a dimer. Two zones of activity stained for LAP were found. The segregation of the two zones (LAP-A and LAP-B) suggested that tow loci control each of both isozymes. Two and three alleles were identified at both loci. GOT-B and LAP-B were found to be tightly linked, showing an average recombination frequency of 12.5 percent. Slight deviation from independent assortment was observed between GOT-B and GOT-C, with recombination frequency of 41 percent.

• Journal of Korean Society of Forest Science
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• v.29 no.1
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• pp.102-114
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• 1976

This experiment was conducted to investigate the percentage of fertile seed in terms of crossabilities and relationships of taxonomic affinities for the ${\times}$ P. rigitaeda of interspecific hybrid, ${\times}$ P. rigida rigitaeda and ${\times}$ P. rigitaeda rigida of backcross hybrids, $F_2$ of ${\times}$ P. rigitaeda and natural hybrid of ${\times}$ P. rigitaeda within Sub-genus Diploxylon of the Genus Pinus. The possibility of establishment of hybrid seed orchard and differentia of hybrids for the purpose of extensive program of reforestation in the future have also been investigated. And, the experimental results obtained are summarized as follows: 1. On the basis of crossabilities as well as on the taxonomic affinities according to the systems of Shaw, Pilger and Duffield, it has been proven that the parental species of those hybrids are of close affinities and range of the fertile hybrid seed production rate was as high as 67-87% in the best hybrid combination (Table 6). 2. Those hybrids seemed to be most promising in the growth perfermance exhibiting 28-80% more volume growth compared to the P. rigida with the statistic significance of 1-5% level (Table 7, 8, 9). And all hybrids exhibit cold hardiness as much as P. rigida except $F_1$ hybrid of ${\times}$ P. rigitaeda and it seems to suggest that the characteristics of cold hardiness were transmitted from the P. rigida. 3. With regard to the anatomical characteristics of needle, the hypoderm is biform in most of the hybrid pines and the characteristics of resin canals are medial in all hybrid. And, the fibrovascular bundles are intermediate of both parent in all hybrid. Therefore it was found to be possible to distinguish the hybrids pines from their parents by the needle characteristics (Table 10). 4. It has been demonstrated that the hybrids pines have a phenolic substance (No. 7) of pale yellow at Rf-0.66, same as P. rigida, but no trace of phenolic substance was observed in the P. taeda. This fact will serve as an important criteria for early identification of hybridity in progeny testing (Table 11). 5. It was found to be possible to distinguish by the starch gel electrophoretic variations banding patterns and staining densities of isoperoxidase in the needles of the hybrids pines from their parents (Fig. 1).

• Korean Journal of Food Science and Technology
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• v.35 no.6
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• pp.1098-1103
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• 2003

Soybeans released proteins when immersed in water at $50{\sim}60^{\sim}C$. We investigated the changes in the characteristics of soybean when soaked in water at different temperatures and studied the electrophoretic properties of soy proteins in recommended Korean soybean varieties after heat treatment. Soybean seeds were heated in soaking water at temperatures of 30, 40, 50, 60, $70^{\circ}C$ for 90 min, and also from 10 to 150min at $60^{\circ}C$. The pH value of the water decreased with heating time at $60^{\circ}C$, and the amount of soluble solids increased with temperature and heating time. The protein concentration of the solution increased with temperature and time. From SDS-PAGE of the proteins in soaking water, we detected two new bands of 16 kDa- and 31 kDa-proteins from the Korean soybean varieties on heat treatment.

• Animal Systematics, Evolution and Diversity
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• v.9 no.2
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• pp.151-170
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• 1993

Morphometric, band-pattern and electrophoretic analysis on Cobitis taenia complex were performed to investigate the morphological and genetic differentiation and to clarify their taxonomic status. Intermediate types of band-pattern (C and D type) were more frequently expressed than that of types of C. t. taenia(type A) and C. t. lutheri (type B). Sexual dimorphism of band-pattern was observed not only in C. t. taenia(type A) and C. t. lutheri(type B). Sexual dimorphism of band-pattern was observed not only in C. t. lutheri but also in C. t. taenia and C. t. striata as well. Discriminant function analysis based on 19 morphological characters shows no significant differences among C. taenia complex. The degree of genic variation of C. t. striata was higher ( =1.48, P=31.2%, HD=0.009) than those of C. t. striata was higher( =1.48, P=31.2%, HD=0.082 and HG=0.009) than those of C. t. lutheri ( =1.37, P=2.7%, HD=0.058 and HG=0.065). The average genetic similarities between C. t. lutheri and C. t. taenia-C. t. striata were S=0.62 and S=0.66 respectively and these values indicate that C. t. tanenia has evolved specific level of differentiation. C. t. striata and C. t. lutheri show subspecifc level of close genetic similarity (S=0.82). Based on the divergent time estimate (Nei, 1975) it is assumed that C. t. tanenia was branched off from the other subspecies about two million years before present (MYBP) and C. t. striata and C. t. lutheri were differentiated about 0.6 MYBP. The use of C. sinesis an the scientific name for the Korean C. t. taenia, proposed by Kim and Lee (1988) seems incorrect since they are quite different in the structure of lamina circularis (Vladycov, 1935), the external morphology and distribution (Cheng and Zheng, 1987) and the chromosome number(Yu et al., 1989). Kim and Lee(1988) also argued that C.t. striata and C. t. lutheri should be treated as distinct species but the present study and other reports (Kim and Lee, 1984; Kim and Yang, 1993) do not support it. We conclude that C. t. taenia is a good species and C. t. striata and C. t. lutheri are subspecific status. Their scientific names should be revised in the future.

• Journal of Life Science
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• v.15 no.3 s.70
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• pp.439-446
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• 2005

Ferritin is known to be the principle iron-storage protein in a wide variety of rganisms. The electro­phoretic mobility and immunological cross-reactivity of dog splenic ferritin were compared with those of horse, bovine, and pig splenic ferritin after isolation using heat treatment, salting out, column chromatography, and ultrafiltration. These isolation methods allowed the recovery of $\~84{\mu}g$ of the ferritin per g of spleen. The iron content in the dog ferritin was $22.7\%$, which appeared to be higher than those in the other mammalian ferritins tested. The electrophoretic mobility of the dog ferritin under nondenaturing conditions was similar to its bovine counterpart, whereas it was more identical to pig and horse ferritins on an SDS-polyacrylamide gel. The molecular weight of the dog ferritin subunit was 19.5 kDa on an SDS-polyacrylarnide gel, and the subunit was unable to bind with iron. The polyclonal anti-dog ferritin raised in rats was able to cross-react with the pig, bovine, and horse ferritins, upon Ouchterlony double immunodiffusiion. A Western blot analysis also revealed that the anti-dog ferritin, which specifically bound with the dog ferritin subunit, could also recognize the horse, bovine, and pig ferritin subunits and the maximum cross-reactivity was exhibited with the pig ferritin subunit, indicating that the dog ferritin is immunochemically more similar to the pig ferritin than its other mammalian counterparts. Accordingly, these results elucidate the biochemical and immunochemical characteristics of dog ferritin that might have a potential to be applied as an oral iron supplement to treat iron deficiency anemia.

• Korean Journal of Food Science and Technology
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• v.15 no.4
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• pp.370-378
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• 1983

Changes of protein and amino acids composition in improvement-Meju inoculated with Aspergillus oryzae were emamined at various time intervals over 6-day test period. To investigate those changes systematically, Disc gel electrophoresis, gel fiteration and amino acid analyzer were used. Following results were obtained; 1. Nitrogen solubility of the soybean meal in $Na_{2}SO_{4},\;MgSO_{4},\;Na_{2}CO_{3},\;NaCl\;and\;Na_{2}HPO_{4}$ solutions of various concetrations were determinated. The salt soluble protein of soybean meal was highly dispersible on 0.4M $Na_{2}SO_{4}$ solution and the extractability of protein was 33%. 2. From the quantitative fractionation of soybean proteins, albumin content (46.0%) was highest followed by globulin (33.9%), glutelin (19.5%) and prolamin (2.4%). During Meju incubation period, albumin and prolamin increased gradually but glutelin decreased. Globulin content was not changed substantially. 3. When albumin was fractionated by Sephadex G-200, the following results were obtained. Soybean albumin showed fraction which was reduced to 3 fraction at 0-day of incubation. The number of fraction, however increased to 8 after 6-day of incubation. 4. Amino acids of albumin in soybean and Meju appeared to be 17 kinds. Glutamic acid and aspartic acid were the highest. In amino acid composition of cooked soybean albumin, arginine, aspartic acid, glutamic acid and glycine remained higher than those of Meju throughout incubation period. 5. The major fraction of albumins from soybean and Meju fractionated by Sephadex G-200 showed 17 kinds of amino acid. Aspartic acid and glutamic acid were the highest. During Meju incubation period, the change of amino acid composition was investigated; threonine, serine, lysine, histidine, alanine, isoleucine, leucine, phenylalanine and $NH_3$ was increased gradually, the others decreased. 6. According to the electrophoretic pattern, soybean protein showed 13 bands which decreased to 3-after cooking. During incubation, those bands increased gradually to 10 bands after 6-days.

• Journal of Animal Science and Technology
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• v.46 no.6
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• pp.887-896
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• 2004

The present study was carried out to investigate the blood markers of Mongolian horses. The blood redcell types and blood protein types(biochemical polymorphisrns) were tested from 19 Mongolian horses by serological and electrophoretic procedure, and their phenotypes and gene frequencies were estimated. The blood group and biochemical polymorphism phenotypes observed with high frequency were $A^{af}$(42.1%), $C^a$(89.5%), $K^-$(84.2%), $U^a$(63.2%), $P^a$(42.1%) $P^-$42.1%), $Q^c$(31.6%) $Q^-$(31.6%), $AL^{AB}$((52.6%), AI$B^K$(89.5%), $ES^1$(63.2%), $GC^F$(78.9%), $HB^BI$1(68.4%), PG$D^F$(84.2%), $TF^{FIR}$(21.1%), $TF^{F2R}$(21.1%)(21.1%), and genotypes $D^{cgm/dghm}$(15.8%), $D^{dghm/dghm}$(15.8%), $D^{ad/dghm}$(10.5%), $D^{ade/dghm}$(10.5%), in Mongolian horses, respectively. Alleles observed with high frequency were $A^a$(0.4211), $C^a$(0.8947), $K^-$(0.8421), $U^a$(0.6316), $P^a$(0.4474), $Q^c$(0.4474), $D^{dghm}$(0.4211), $AL^B$(0.6579), $AIB^K$(0.9211), $ES^I$(0.7895), $GC^F$(0.8947), $HB^{BI}$(0.7895), $PGD^F$(0.8421) and $TF^R$(0.3421) in Mongolian horses. These results present basic information for estimating the genetic relationships between the Korean native horse, and developing a system for parentage verification and individuals identification in Mongolian horse.