• Title/Summary/Keyword: Gal2

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$\beta$-Subunit 94~96 Residues of Tethered Recombinant Equine Chorionic Gonadotropin are Important Sites for Luteinizing Hormone and Follicle Stimulating Hormone like Activities

  • Park, Jong-Ju;JarGal, Naidansuren;Yoon, Jong-Taek;Min, Kwan-Sik
    • Reproductive and Developmental Biology
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    • v.34 no.1
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    • pp.33-40
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    • 2010
  • Equine chorionic gonadotropin (eCG) is a heavily glycosylated glycoprotein composed of non-covalently linked $\alpha$- and $\beta$-subunits. To study the function and signal transduction of tethered recombinant-eCG (rec-eCG), a single chain eCG molecule was constructed, and the rec-eCG protein was prepared. In this study, we constructed 5 mutants (${\Delta}1$, ${\Delta}2$, ${\Delta}3$, ${\Delta}4$, and ${\Delta}5$) of rec-eCG using data about known glycoprotein hormones to analyze the role of specific follicle stimulating homone (FSH)-like activity. Three amino acids of certain specific sites were replaced with alanine. The expression vectors were transfected into CHO cells and subjected to G418 selection for 2~3 weeks. The media were collected and the quantity of secreted tethered rec-eCGs was quantified by ELISA. The LH- and FSH-like activities were assayed in terms of cAMP production by rat LH/CG and rat FSH receptors. Then, the metabolic clearance rate analyzed by the injection of rec-eCG (5 IU) into the tail vein was analyzed. The mutant eCGs (${\Delta}l$, ${\Delta}4$, and ${\Delta}5$) were transcripted, but not translated into proteins. Rec-eCG A2 was secreted in much lower amounts than the wild type. Only the rec-eCG ${\Delta}3$ ($\beta$-subunit: $Gln^{94}-Ile^{95}-Lys^{96}{\rightarrow}Ala^{94}-Ala^{95}-Ala^{96}$) was efficiently secreted. Although activity is low, its LH-like activity was similar to that of tethered $eCG{\beta\alpha}$. However, the FSH-like activity of rec-$eCG{\beta\alpha\Delta}3$ was completely flat. The result of the analysis of the metabolic clearance rate shoed the persistence of the mutant in the blood until 4 hours after the injection. After then, it almost disappeared at 8 hours. Taken together, these data suggest that 94~96 amino acid sequences in eCG $\beta$-subunit appear to be of utmost importance for signal transduction of the FSH receptor.

Quality Properties of Baechu kimchi treated with Black Panax ginseng Extracts during Fermentation at Low Temperature (흑삼추출액을 첨가한 배추김치의 저온 저장 중의 품질 특성)

  • Mo, Eun-Kyoung;Kim, Seung-Mi;Yun, Beom-Sik;Yang, Sun-A;JeGal, Sung-A;Choi, Young-Sim;Ly, Sun-Yung;Sung, Chang-Keun
    • Food Science and Preservation
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    • v.17 no.2
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    • pp.182-189
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    • 2010
  • To develop a new functional kimchi with cognition-enhancing properties, black Panax ginseng extract (0.5-5%, w/w) was added to a baechu kimchi preparation and the mixture stored at $4^{\circ}C$ for 30 days. Compared with control kimchi, the L values of ginseng-treated material were significantly decreased, but the a and b values were increased. The hardness value of ginseng-treated kimchi was significantly higher than that of control material from the $20^{th}$ day of storage. The edibility period of baechu kimchi treated with ginseng was prolonged by approximately 15 days compared with control kimchi. This resulted from decreases in the numbers of lactic acid bacteria and yeasts during the final stages of fermentation in ginseng-treated material. Inhibition of acetylcholinesterase activity by ginseng-treated kimchi was 2-fold higher than that of control material. A strong ginseng flavor and a bitter taste were evident in kimchi treated with 5% (w/w) ginseng, and sensory quality was thus decreased compared with control material. It was concluded that an appropriate concentration of black ginseng extract was 3% (w/w) in preparation of kimchi with a cognition-enhancing effect.

Studies on the Content of Lectin in Korean Mistletoe according to the Host Tree Species and Characterization for Its Application to the Quality Control (한국산 겨우살이 숙주별 렉틴 함량과 지표물질로서의 특성 조사)

  • Kim, Inbo;Yoon, Taek Joon;Park, Choon Ho;Lee, Woo Kyoung;Lee, So Hee;Kim, Jong Bae
    • The Korean Journal of Food And Nutrition
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    • v.28 no.6
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    • pp.1090-1097
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    • 2015
  • Traditionally, mistletoe is known as an effective anti-cancer medicinal plant, and lectin is recognized as a major component with cytotoxic and immuno-stimulant activity in mistletoe. A Korean mistletoe lectin (KML) has specificity to galactose and galactosamine and is distinguish from European mistletoe lectin (EML). When we examined the concentration of lectin in mistletoe originated from five different types of host trees, the result indicate that the lectin concentration is variable depending on the host tree. Noticeably, mistletoe from chestnut tree contains ten folds higher lectins than that of an oak tree. We also tested the concentration of KML and crude extract (KM-110) of Korean mistletoe that shows 90% cytotoxicity in L5178Y-ML25 lymphoma cell. In addition, the cells show 90% and 70% viability by the treatment of two neutralizing antibodies of KML, 9H7-D10 and 8B11-2C5 neutralization effect with two monoclonal antibodies of KML, 9H7-D10 and 8B11-2C5. Therefore, the result expected that the mistletoe contain some other cytotoxic components except lectin. Finally, the production of $TNF-{\alpha}$ and IL-6 by RAW 264.7 cells stimulated with lectin free-crude extract (LFKM-110) following neutralization by 9H7-D10 monoclonal antibody shows higher than that of lectin containing-crude extract (KM-110). These results suggest that the Korean mistletoe lectin ha a great potential to be developed as therapeutic agent of cancer.

Characterization of Bacillus licheniformis KJ-9 Isolated from Soil (토양으로부터 분리한 Bacillus licheniformis KJ 9의 특성)

  • Seo, Dong-Cheol;Ko, Jeong-Ae;Gal, Sang-Won;Lee, Sang-Won
    • Journal of Life Science
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    • v.20 no.3
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    • pp.403-410
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    • 2010
  • In order to produce high-quality fermenting composts, a microorganism was isolated from the natural world. The bacterium has not only in high enzyme activities but also had good antimicrobial activities against phytopathogenic microorganisms. Its cultivating characteristics were then investigated. Bacterium KJ-9, which contains high CMCase, protease and chitinase activities and excellent antimicrobial activities against phytopathogenic microorganisms, was separated from leaf mold and identified as Bacillus licheniformis by two methods: Bergey's Manual of Systematic Bacteriology and API 50 CHL Carbohydrate Test Kit (Bio Merieux, France) using an ATB (Automated Identification) computer system (Bio Merieux, France). Optimal medium for cultivation of B. licheniformis was 2% soluble starch as a carbon source, 0.5% yeast extract as a nitrogen source and 0.05% $MgSO_4{\cdot}7H_2O$. Optimal growth conditions of pH, temperature and shake speed were pH 7.0, $50^{\circ}C$ and 180 rpm, respectively. Culture broth of B. licheniformis KJ-9 cultured for 36~60 hr was effective in fungicidal activities against plant pathogens including Botrytis cinerea, Corynespora cassicola, Fusarium oxysporum, and Rhizoctonia solani.

Central Neural Pathways Projecting to the Acupoints B62 and K6 Using the Neural Tracer (신경추적자를 이용한 신맥과 조해에서 투사되는 신경원의 표지부위에 대한 연구)

  • Kim, Su-Hyun;Lee, Chang-Hyun;Yuk, Sang-Won;Lee, Kwang-Gyu;Lee, Tae-Young;Lee, Sang-Ryoung
    • Journal of Acupuncture Research
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    • v.18 no.2
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    • pp.51-66
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    • 2001
  • The purpose of this morphological studies was to investigate the central neural pathway projecting to the acupoints $B_{62}$ and $K_6$ using the neuroanatomical method following injection of transsynaptic neurotropic virus, pseudorabies virus(PRV-Ba and PRV-Ga) into the $B_{62}$ and $K_6$. After survival times of 96 hours following injection into the twenty rats with PRV-Ba(Bartha strain) and PRV-Ga(Bartha strain, ${\beta}$-galacidodase insertion). They were perfused, and their spinal cord and brain were frozen sectioned($30{\mu}m$). These sections were stained by X-gal histochemical and PRV immunohistochemical staining method, and observed with light microscope. The results were as follows : 1. In spinal cord, overlaped PRV-Ba and PRV-Ga labeled neurons projecting to the $B_{62}$ and $K_6$ were founded in thoracic, lumbar and sacral spinal segments. In thoracic spinal segments, Densely labeled areas were founded in lamina IV, V, VII(intermediolateral nucleus) and X areas. In lumbar segemnts, labeled areas were founded in lamina II, IV, V and X areas. In sacral spinal segments, labeled areas were founded in lamina IV, V and VI areas. 2. In brain, overlaped PRV-Ba and PRV-Ga labeled neurons projecting to the $B_{62}$ and $K_6$ were founded in the $A_1$ noradrenalin cells/$C_1$ adrenalin cells/caudoventrolateral reticular nucleus, rostroventrolateral reticular nuclens, nucleus tractus solitarius, area postrema, raphe obscurus nucleus, raphe paltidus nucleus, raphe magnus nucleus, lateral paragigantoceltular nucleus, lateral rcticular nucleus, gigantocellular nucleus, locus coeruleus, subcoeruleus nucleus, motor trigeminal nucleus, Kolliker-Fuse nucleus, $A_5$ cell group, central gray matter, oculomotor nerve, paraventricular hypothalamic nucleus, median eminence, amygdaloid nucleus, frontal cortex, forelimb area, hindlimb area, 1, 2 areas of parietal cortex and granular and agranular cortex. This results were suggest that overlaped PRV-Ba and PRV-Ga labeled areas projecting to the $B_{62}$ and $K_6$ may be related to the emotional relay pathway in the central autonomic center.

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Carcinogenicity and mutagenicity of heterocyclic amines in transgenic models

  • Ryu D.Y.
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2000.11a
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    • pp.45-67
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    • 2000
  • 2-Amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) is a mutagenic and carcinogenic heterocyclic amino found in cooked meat. The in vivo mutagenicity and hepatocarcinogenicity of MeIQx were examined in mice harboring the lacZ mutation reporter gene ($Muta^{TM}$ Mice) and bitransgenic mice over-expressing the c-myc oncogene. C57B1/$\lambda$lacZ and bitransgenic c-myc (albumin promoter)/$\lambda$lacZ mice were bred and weaned onto an AIN-76 based diet containing $0.06\%$ (w/w) MeIQx or onto control diet. After 30 weeks on diet, only male bitransgenic mice on MeIQx developed hepatocellular carcinoma ($100\%$ incidence) indicating that there was synergism between c-myc over-expression and MeIQx. By 40 weeks, hepatic tumor incidence was $100\%$ ($17\%$) and $44\%$ ($0\%$) in male c-myc/$\lambda$lacZ and C57B1/$\lambda$lacZ mice given MeIQx (or control) diet, respectively, indicating that either MeIQx or c-myc over-expression alone eventually induced hepatic tumors. At either time point, mutant frequency in the lacZ gene was at least 40-fold higher in MeIQx-treated mice than in control mice of either strain. These findings suggest that MeIQx-induced hepatocarcinogenesis is associated with MeIQx-induced mutations. Elevated mutant frequency in MeIQx-treated mice also occurred concomitant with the formation of MeIQx-guanine adducts as detected by the $^{32}P$-postlabeling assay. Irrespective of strain or diet, sequence analysis of the lacZ mutants from male mouse liver showed that the principal sequence alteration was a single guanine-base substitution. Adenine mutations, however, were detected only in animals on control diet. MeIQx-fed mice harboring the c-myc oncogene showed a l.4-2.6-fold higher mutant frequency in the lacZ gene than mice not carrying the transgene. Although there was a trend toward higher adduct levels in c-myc mice, MeIQx-DNA adduct levels were not significantly different between c-myc/$\lambda$lacZ and C57B1/$\lambda$lacZ mice after 30 weeks on diet. Thus, it appeared that factors in addition to MeIQx-DNA adduct levels, such as the enhance rate of proliferation associated with c-myc over-expression, may have accounted for a higher mutant frequency in c-myc mice. In the control diet groups, the lacZ mutant frequency was significantly higher in c-myc/$\lambda$lacZ mice than in 057B1/$\lambda$1acZ mice. The findings are consistent with the notion that c-myc over-expression is associated with an increase in mutagenesis. The mechanism for the synergistic effects of c-myc over-expression on MeIQx hepatocarcinogenicity appears to involve an enhancement of MeIQx-induced mutations.

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Study of Composite channel Structure of Metamorphic HEMT for the Improved Device Characteristics (기존의 MHEMT와 InP 합성 채널 MHEMT의 소자의 항복 특성 분석 및 비교 연구)

  • Choi, Seok-Gyu;Baek, Yong-Hyun;Han, Min;Bang, Seok-Ho;Yoon, Jin-Seob;Rhee, Jin-Koo
    • Journal of the Institute of Electronics Engineers of Korea SD
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    • v.44 no.12
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    • pp.1-6
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    • 2007
  • In this study, we have performed the channel modification of the conventional MHEMT (metamorphic high electron mobility transistor) to improve the breakdown characteristics. The Modified channel consists of the InxGal-xAs channel and the InP sub channel instead of the InxGa1-xAs channel. Since InP has the lower impact ionization coefficient in comparison with In0.53Ga0.47As, we have adopted the InP-composite channel in the modified MHEMT. We have investigated the breakdown mechanism and the RF characteristics for the conventional and the InP- composite channel MHEMTs. From the measurement results, we have obtained the enhanced on and off-state breakdown voltages of 2.4 and 5.7 V, respectively. Also, the increased RF characteristics have brought about the decreased output conductance for the InP-composite channel MHEMT. The cut-off frequency (fT) and the maximum oscillation frequency (fmax) for the InP-composite Channel MHEMT were 160 GHz and 230 GHz, respectively. It has been shown that the InP-composite channel MHEMT has the potential applications for the millimeter wave power device.

Effects of antimony addition on growth of InGaN nano-structures by mixed-source HVPE (혼합소스 HVPE 방법에 의한 InGaN 나노구조의 성장에 있어서 Sb 첨가의 영향)

  • Ok, Jin-Eun;Jo, Dong-Wan;Jeon, Hun-Soo;Lee, Ah-Reum;Lee, Gang-Suok;Cho, Young-Ji;Kim, Kyung-Hwa;Chang, Ji-Ho;Ahn, Hyung-Soo;Yang, Min
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.20 no.3
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    • pp.113-116
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    • 2010
  • We report on the growth and characteristics of the structural and optical properties of InGaN nano-structures doped with antimony (Sb) as a catalyst. The use of catalyst has been explored to modify the growth and defect generation during strained layer heteroepitaxial growth. We performed the growth of the InGaN nano-structures on c-sapphire substrates using mixed-source hydride vapor phase epitaxy (HVPE). The characteristic of samples was measured by scanning electron microscope (SEM) and photoluminescence (PL). The aligning direction of c-axis of the InGaN nano-structures was changed from vertical to parallel or inclined to the surface of substrates when the Sb was added as a catalyst. The indium composition was estimated about 3.2% in both cases of with or without the addition of Sb in the InxGal-xN structures. From the results of InGaN nano-structures formed with the addition of Sb, we can expect the performance of optical devices would be more improved by reduced piezo-electric field if we use the InGaN nano-structures of which c-axes are aligned parallel to the substrates as an active layer.

The Study on the Aluminum Content in Fishes Caught from Several Areas of the West Coast in Korea (수산식품 중 노인성 퇴행성 질환과 관련된 알루미늄 함량 및 그에 따른 수산식품의 이용방안에 관한 연구)

  • 김애정
    • Journal of the East Asian Society of Dietary Life
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    • v.7 no.4
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    • pp.512-518
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    • 1997
  • This study was performed to determine the amount of aluminum, which is one of the factors of Alzheimer's disease, In some fishes caught from some areas of the west coast in Korea. The 46 aquatic products were composed of fishes, molluscs, and salt-fermented products (jeot-gal). The 24 fishes were Hickory shad, Gobies, Pomfref, Atkafish, Flounder, Jambeng-ie Monk fish, Yellow hair tail, Mackerel, Bartailed flathead, Alaska pollack, Brown croaker, Eel, Fine-spotted flounder, Black spotted grouper, Sea-eel, Pacific saury, Areliscus honaleus, Small boil-dried anchovy, Croaker, Hair tail, Sea bream genuine, Motleystrip rainbowfish, and Bastard halibut. The 15 Molluscs were Whip-arm octopus, Sea arrow, Common squid, Han chi, Cuttle fish, Turban shell, Pond snail, Orient calm, Surf calm, Butter calm, Crib shell, Oyster, Egg cockle, Little neck calm, and Arkshell. The 7 salt-fermented products were salt-fermented Shrimp, Little neck, Oyster, Shad, Gonjeng-ie, Hqangsegi, and Squid. All of them were ashed with 5$m\ell$ HNO$_3$ and then with 10$m\ell$ ternary solution (HNO$_3$ : H$_2$SO$_4$ : HClO$_4$= 10 : 1 : 4). After ashing of the samples, the aluminum amount were measured by ICP. The aluminum amount of molluscs was significantly higher than that of fishes and salt-fermented products(p<0.01). The aluminum amount of Orient calm and Healak in molluscs were 827.70, 812.55ppm, respectively, which were the most amounts compared nth that of the other samples. But the aluminum amounts of Bartailed flathead and Sea bream, genuine In fishes were 0.98, 0.97ppm, respectively, which were the least amounts compared with that of other samples. This study was limited within 46 aquatic samples, therefore I hope there will be wider efforts to determine about auminum amount in broade range of aquatic foods for the prevention of Alzheimer's disease.

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Cloning and High Expression of Nattokinase Gene from Bacillus subtilis BB-1 (Bacillus subtilis BB-1으로부터 나토키나아제 유전자 크로닝 및 대량발현)

  • Lee Young-Hoon;Lee Sung-Ho;Park Ki-Hoon;Choi Young-Ju;Jeong Yong-Kee;Gal Sang-Wan
    • Journal of Life Science
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    • v.16 no.2 s.75
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    • pp.274-281
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    • 2006
  • A fibrinolytic enzyme gene was isolated from Bacillus subtilis BB-1 by PCR method. Primers for PCR cloning were designed according to pre-identified gene for fibrinolytic enzymes from B. subtilis. The primer sequences were 5'-CGG ATC CGT GAG AGG CAA AAA GGT G-3' and 5'-TGA ATT CTT AAT GTG CTG CTG CTT GTC C-3' as concensus sequences of the fibrinolytic genes of Bacillus species. The PCR product was 1,145 bp and the sequence homology was 99% with nattokinase gene isolated from Japanese natto. The cloned fibrinolytic gene was reconstructed in Bacillus-E. coli shuttle vector, pEB for bulk-production. The fibrinolytic enzyme was purified by FPLC from the cloned B. subtilis 168. The optimum pH and temperature of the enzyme were 7.0 and $35^{\circ}C$, respectively. The fibrinolytic enzyme did not show any activity toward to skim milk, gelatin, casein and blood agar plate. The enzyme specific polyclonal antibody was prepared in rabbit for further assays such as detection of the gene expression in plant cells. This means that the enzyme may be used for health-care such as thrombosis without any hamful effects in the blood vessel.