• Title/Summary/Keyword: BL52

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Studies on the Sexing of Bovine Embryo by the Chromosomal Analysis and H-Y Antibody (염색체 분석 및 H-Y 항체처리에 의한 우수정란의 성판별에 관한 연구)

  • 고광두;양부근;정희태;박연수;김정익
    • Journal of Embryo Transfer
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    • v.3 no.1
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    • pp.48-52
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    • 1988
  • 우수정란의 이식전 성판별이 관한 연구를 수행하기 위하여 GTH와 PGF$_2$$\alpha$투여에 대한 난소반응과 회수난자의 발유단계별 동결융해후 생존성을 조사하였으며, 이식전 수정라느이 성판별을 위하여 H-Y항체 처리후 정상발육 난자의 염색체를 분석하여 다음과 같은 결과를 얻었다. 웅성 비장세포(male, spleen cells)를 면역원으로 mouse와 rat에 투여, 항혈청의 항체를 확인한 결과 mouse에서는 C57 BL계통과 rat에서는 DonRyu 계통이 항체생산능력이 우수하였다. 공란우 87두에 hormone(2500IU PMSG, 25mg PGF$_2$alpha)처리하여 평균 57.8%의 채란유과 두당 4.9개의 난자가 회수되었으며, 전체회수란자(427개)중 moula(162개)와 blastocyst(190개)의 정상발육란자는 82.4%였다. 동결융해후 회수된 난자 (312개)중, 형태적으로 정상인 난자(241개)의 비율은 77.2% 발육단계별 성적은 blastocyst(83.4%)가 morula(71.0%)보다 우수하였다. 항체와 보체(Guinea pig serum)로 처리된 82개의 morula중 15개(18.3%)가 blastocyst로 발육되어 이중 5개(33.3%)가 성이 판별되었으며, 모두 xx형 성염색체를 갖는 자성수정란으로 판명되었다.

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Molecular Cloning of Seven-band Grouper (Epinephelus septemfasciatus) Growth Hormone cDNA and Its Expression in Escherichia coli

  • Lee Jehee;Munasinghe Helani;Song Choon Bok
    • Fisheries and Aquatic Sciences
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    • v.6 no.3
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    • pp.116-124
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    • 2003
  • Isolation and cloning of seven-band grouper (Epinephelus septemfasciatus) growth hormone cDNA from pituitary gland revealed an open reading frame of 612 bp coding for a pre-growth hormone of 204 amino acids with a 17 amino acid putative signal peptide. Deduced amino acid sequence showed that there was one possible N-glycosylation site at $Asn^{l84}$ and four cysteine residues $(Cys^{52},\;Cys^{160},\;Cys^{177},\;Cys^{185})$ on t e same positions as in some other species where they were involved in the stabilization of the tertiary structure. The seven-band grouper growth hormone (sbgGH) presented a $99.5\%$ amino acid sequence identity with the growth hormone of Epinephelus coioides and contained the conserved hormone domain region. Comparison of growth hormone sequences from evolutionarily diverse species revealed 25 amino acid residues conserved in jawless fishes to modern mammals. It also revealed an evolutionary trend to retain the same polypeptide sequence even in the distantly related animals while allowing alterations to occur in polypeptides of the closely related species. In order to create a recombinant system to produce high levels of the growth hormone, it was expressed in Escherichia coli (BL21) cells. The gel analysis revealed theoretically expected molecular weights for both mature and pre-sbgGHs.

Ethylene gas adsorption capacity and preserving effect of fruit freshness of Charcoal-fiberboard by wet forming process (습식공법으로 제조한 목탄-목재섬유복합재료의 에틸렌가스 흡착력과 과일 신선도 유지 효과)

  • Lee Hwa Hyoung;Kim Gwan Eui
    • Journal of the Korea Furniture Society
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    • v.14 no.1
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    • pp.1-9
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    • 2003
  • This research was carried out for packing materials and building materials to examine thylene gas adsorption and effect of keeping fruit fresh of wet formed charcoal-fiber mposite made from defibrated fiber of Pinus densiflora Sieb. et Zucc. and white charcoal from uercus variabilis Bl.(wood fiber: charcoal=8:2, 6:4, 4:6, 2:8), with/without phenol formaldehyde resin(PF, Non volatile content:$52\%$, resin content $1,3,5\%$). The results are summarized as follows: 1. The higher the charcoal content, the more the ethylene gas adsorption. At the same mixing ratio of fiber to charcoal, $\#100-200$ of charcoal particle size gave the better reslts than $\#60-100$. 2. Adding PF into the charcoal fiber composite decreased the capacity of ethylene gas adsorption but there was no significant difference until $5\%$ adding amount of PF. 3. For keeping fruit fresh for a long time, Charcoal fiber composite was $66\%$ longer than control. The higher the white charcoal content, the longer fresh time.

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Unfolded Histidine-Tagged Protein is Immobilized to Nitrilotriacetic Acid-Nickel Beads, But Not the Nickel-Coated Glass Slide

  • Cho Min-Ho;Ahn Sun-Young;Park Heon-Yong
    • Genomics & Informatics
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    • v.4 no.3
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    • pp.133-136
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    • 2006
  • The adsorption of proteins on the surface of glass slides is essential for construction of protein chips. Previously, we prepared a nickel-coated plate by the spin-coating method for immobilization of His-tagged proteins. In order to know whether the structural factor is responsible for the immobilization of His-tagged proteins to the nickel-coated glass slide, we executed a series of experiments. First we purified a His-tagged protein after expressing the vector in E. coli BL21 (DE3). Then we obtained the unfolding curve for the His-tagged protein by using guanidine hydrochloride. Fractions unfolded were monitored by internal fluorescence spectroscopy. The ${\Delta}G_{H20}$ for unfolding was $2.27kcal/mol{/pm}0.52$. Then we tested if unfolded His-tagged proteins can be adsorbed to the nickel-coated plate, comparing with $Ni^{2+}-NTA$ (nitrilotriacetic acid) beads. Whereas unfolded His-tagged proteins were adsorbed to $Ni^{2+}-NTA$ beads, they did not bind to the nickel-coated plate. In conclusion, a structural factor is likely to be an important factor for constructing the protein chips, when His-tagged proteins will immobilize to the nickel-coated slides.

Antioxidant and tyrosinase inhibitory activity of Hypsizygus marmoreus (brown cultivar) methanol extracts (갈색 느티만가닥버섯 메탄올 추출물의 항산화 및 tyrosinase 저해 효과)

  • Kim, Su Cheol;Ryu, Han Min;Jung, Sung Mi;Lee, Young Hyun;Kim, Hye Soo;Kim, Jong Ok;Cho, Young Un;Cho, Soo Jeong
    • Journal of Mushroom
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    • v.11 no.4
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    • pp.254-260
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    • 2013
  • The objective of this study was to evaluate antioxidant effect and tyrosinase inhibitory activity of methanol extracts from Hypsizygus marmoreus. The Hypsizygus marmoreus was divided into two parts (pileus and stipe) and extracted with methanol. Total polyphenolics and flavonoids in the methanol extracts were measured by spectrophotometric methods and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activities have been determined for antioxidant activities. The total polyphenolics and flavonoids contents of methanol extract of the pileus were higher than methanol extract of the stipes. The total polyphenolics contents in methanol extracts of the pileus and stipes were 8.7 ug/mg and 5.6 ug/mg, respectively. The total flavonoids contents in methanol extracts of the pileus and stipes were 2.8 ug/mg and 1.4 ug/mg, respectively. The tyrosinase inhibitory activity was proportional to concentration of methanol extract. The tyrosinase inhibitory activity of the methanol extract (200 mg/ml) of pileus (66.9%) and stipe (57.97%) was lower than those of positive control 2% arbutin. The DPPH radical scavenging activity of the methanol extract (20 mg/ml) of pileus and stipes was 52.55% and 30.35%, respectively. Moreover, the effects of methanol extarcts on cell proliferation of B16BL6 mouse melanoma cells were investigated using WST-1 assay (4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulphonate) and B16BL6 mouse melanoma cells treated with methanol extract of 200-2,000 ug/ml were higher proliferation rate than those of 0.04% adenosine.

Pilot Research about Influential Factors and Efficacy Judgement of Acupoint Sticking in Dog-Days (삼복첩(三伏貼)의 영향 요인 및 임상 효과 판정에 관한 예비 연구)

  • Lee, Sun-Haeng;Kim, Cho-Young;Chang, Gyu-Tae
    • The Journal of Pediatrics of Korean Medicine
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    • v.25 no.1
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    • pp.72-81
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    • 2011
  • Backgrounds: Sticking warm and hot herbs to acupoint in Dog-Days where the strongest Yangqi takes place, Acupoint sticking in Dog-Days is one of Oriental medical treatment to cure winter diseases in the summer. This treatment is widespread in China and Taiwan, and is becoming popular in South Korea nowadays. Objectives: The research has been conducted to evaluate the influencial factors and efficacy in Acupoint sticking in Dog-Days. Methods: We visited one kindergarten in Dog-Days of 2010, and attached pills which are made of Sinapis Semen(白芥子), Corydalis Tuber(玄胡索), Asari Herba Cum Radix(細辛), Juice of Zingiber Officinale(薑汁) to BL-13(肺兪), BL-15(心兪), BL-17(膈兪) of 56 children for 4-6 hours(34 boys, 22 girls, $52.64{\pm}16.85$ months, 19-78 months). Exacerbating effect or non-significant changes in respiratory symptoms before and after the treatment were ified.. The influentialfactors of efficacy were analyzed by Pearson's $x^2$-test and Fisher's exact test. In addition, the efficacy and satisfaction among once, twice, and three times treatment groups were verified by ANOVA. Results: 27 children (48.2%) were effective, but efficacy felt by parents is 45 cases (80.4%). Sex, perspiration, stool, urine, sleep, cold of hands and feet, cold sensitivity, and frequency of Acupoint sticking in Dog-Days has no relation to efficacy(p>0.05). However, the efficacy is poor in 1 and 2 age(p=0.030). Difference among efficacy of once, twice, and three times treatment groups is not significant(p=0.069), whereas there are linear association in frequency of Acupoint sticking in Dog-Days and efficacy(p=0.030). 36 parents (64.3%) are satisfied with Acupoint sticking in Dog-Days, and we observed significant difference among once, twice, three times treatment groups(p=0.041). 19 parents (33.9%) had been known about Acupoint sticking in Dog-Days, but 53 parents (94.6%) wanted repetition, and 41 parents (73.2%) intended to recommend it to acquaintance. Side effects are minor itching in two cases. Twenty seven participants showed positive effect, and forty five people (80.4%) felt beneficial. Sex, perspiration, stool, urine, sleep, cold hands and feet, sensitivity in cold, and frequency of Acupoint sticking in Dog-Days has no relation to the efficacy(p>0.05). Though the efficacy was poor in one and two year old infants, it was linearly correlated with the frequency count and responses from the treatment (p=0.030). Thirty six participants (64.3%) satisfied with the treatment, and so did the first, second, and third participant groups (p=0.041). Even though, only nineteen participants (33.9%) were familiar with Acupoint sticking in Dog-Days, fifty three participants (94.6%) preferred to repeat the treatment, and 41 parents were preferable to recommend the treatment to others. Side effects of minor itching has been reported in a couple cases. Conclusions: Acupoint sticking in Dog-Days is effective in children older than three year old. The effect would be greater when going through the treatment three times. Based on significant satisfaction, intention of repetition and recommendation, strong promotion on Dog-Days are needed to make a better use of Acupoint sticking in Dog-Days for the effective treatment.

Anti-diabetic effect and mechanism of Korean red ginseng extract in C57BL/KsJ db/db mice

  • Yuan, Hai-Dan;Shin, Eun-Jung;Chung, Sung-Hyun
    • Proceedings of the Ginseng society Conference
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    • 2007.12a
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    • pp.57-58
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    • 2007
  • Purpose: Ginseng is a well-known medical plant used in traditional Oriental medicine. Korean red ginseng (KRG) has been known to have potent biological activities such as radical scavenging, vasodilating, anti-tumor and anti-diabetic activities. However, the mechanism of the beneficial effects of KRG on diabetes is yet to be elucidated. The present study was designed to investigate the anti-diabetic effect and mechanism of KRG extract in C57BL/KsJ db/db mice. Methods: The db/db mice were randomly divided into six groups: diabetic control group (DC), red ginseng extract low dose group (RGL, 100 mg/kg), red ginseng extract high dose group (RGH, 200 mg/kg), metformin group (MET, 300 mg/kg), glipizide group (GPZ, 15 mg/kg) and pioglitazone group (PIO, 30 mg/kg), and treated with drugs once per day for 10 weeks. During the experiment, body weight and blood glucose levels were measured once every week. At the end of treatment, we measured Hemoglobin A1c (HbA1c), blood glucose, insulin, triglyceride (TG), adiponectin, leptin, non-esterified fatty acid (NEFA). Morphological analyses of liver, pancreas and white adipose tissue were done by histological observation through hematoxylin-eosin staining. Pancreatic islet insulin and glucagon levels were detected by double-immunofluorescence staining. To elucidate an action of mechanism of KRG, DNA microarray analyses were performed, and western blot and RT-PCR were conducted for validation. Results: Compared to the DC group mice, body weight gain of PIO treated group mice showed 15.2% increase, but the other group mice did not showed significant differences. Compared to the DC group, fasting blood glucose levels were decreased by 19.8% in RGL, 18.3% in RGH, 67.7% in MET, 52.3% in GPZ, 56.9% in PIO-treated group. With decreased plasma glucose levels, the insulin resistance index of the RGL-treated group was reduced by 27.7% compared to the DC group. Insulin resistance values for positive drugs were all markedly decreased by 80.8%, 41.1% and 68.9%, compared to that of DC group. HbA1c levels in RGL, RGH, MET, GPZ and PIO-treated groups were also decreased by 11.0%, 6.4%, 18.9%, 16.1% and 27.9% compared to that of DC group, and these figure revealed a similar trend shown in plasma glucose levels. Plasma TG and NEFA levels were decreased by 18.8% and 16.8%, respectively, and plasma adiponectin and leptin levels were increased by 20.6% and 12.1%, respectively, in the RGL-treated group compared to those in DC group. Histological analysis of the liver of mice treated with KRG revealed a significantly decreased number of lipid droplets compared to the DC group. The control mice exhibited definitive loss and degeneration of islet, whereas mice treated with KRG preserved islet architecture. Compared to the DC group mice, KRG resulted in significant reduction of adipocytes. From the pancreatic islet double-immunofluorescence staining, we observed KRG has increased insulin production, but decreased glucagon production. KRG treatment resulted in stimulation of AMP-activated protein kinase (AMPK) phosphorylation in the db/db mice liver. To elucidate mechanism of action of KRG extract, microarray analysis was conducted in the liver tissue of mice treated with KRG extract, and results suggest that red ginseng affects on hepatic expression of genes responsible for glycolysis, gluconeogenesis and fatty acid oxidation. In summary, multiple administration of KRG showed the hypoglycemic activity and improved glucose tolerance. In addition, KRG increased glucose utilization and improved insulin sensitivity through inhibition of lipogenesis and activation of fatty acid $\beta$-oxidation in the liver tissue. In view of our present data, we may suggest that KRG could provide a solid basis for the development of new anti-diabetic drug.

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A Clinical Study of Bee Venom Acupuncture Therapy on Shoulder Pain Patients in Stroke Sequelae (중풍후견증(中風後遣症)으로 인한 견비통(肩譬痛) 환자(患者)의 봉약침치료(蜂藥鍼治療)에 대한 임상적(臨床的) 고찰(考察))

  • Lee, Dae-Yong;Yeom, Seung-Chul;Kim, Do-Ho;Kim, Dae-Joong;Lee, Geon-Mok
    • Journal of Acupuncture Research
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    • v.23 no.4
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    • pp.69-80
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    • 2006
  • Objectives : The occurrence of shoulder pain after attack of stroke varies from 15% to about 80% in patients. Hemiplegic shoulder pain has been shown to affect stroke outcome in a negative way that it interferes with recovery after a stroke. These following processes have been all postulated as causes of a shoulder pain: glenohumeral subluxation, spasticity, impingement, soft tissue trauma, glenohumeral capsulitis, shoulder hand syndrome. And stroke patients may suffer from pain caused by stroke itself(central post-stroke pain). The aim of this study is to investigate the effectiveness of Bee venom therapy for shoulder pain in stroke patients. Methods : To evaluate the effectiveness of Bee Venom Acupuncture Therapy, 40 patients were allocated into control and treatment group. They were monitored for 3 weeks and followed up with VAS score(with the interval of Initial(YAS1)), 1 week later(VAS2), 2 weeks later(VAS3), 3 weeks later(VAS4), Motor Grade and Passive ROM. Results : VAS score decrease in treatment group compared to control group. Bee Venom Acupuncture Therapy seems to decrease hemiplegic shoulder pain and this effect was statistically significant after 3 weeks. Therefore this therapy could be recommended for the treatment of patients with shoulder pain after stroke and further extensive clinical studies are expected. Conclusion : We suggest that GDS oral administration and electro-acupuncture at $BL_{52}$ & $GB_{39}$ are available for prevention and curing about the postmenopausal osteoporosis.

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The Effect of KKBT in Papain-Induced Osteoarthritis Mice Models (건강부자탕(乾薑附子湯)이 papain으로 유도(誘導)된 골관절염(骨關節炎) 생쥐 모델에 미치는 영향(影響))

  • Sung, Yeong-Suk;Choi, Hak-Joo;Oh, Jeong-Min;Ji, Joong-Gu;Bak, Ji Won;Kim, Dong-Hee
    • Journal of Haehwa Medicine
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    • v.21 no.1
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    • pp.33-52
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    • 2012
  • To evaluate the efficacy of Keonkangbujatang (KKBT) in osteoarthritis treatments, C57BL/10 mice were treated with papain to induce osteoarthritis, and anti-arthritic effects were measured. To ensure safety of the KKBT sample, ALT, AST, BUN, and creatinine levels were measured, and they were all within the normal range. Based on the fact that suppression of inflammatory cytokines leads to the improvement of arthritis, IL-1b, IL-6, TNF-a, and MCP-1 production levels were measured. The cytokines were significantly decreased in serum. Also, mRNA levels of IL-1b, IL-6, and iNOS-II were significantly decreased in joint tissues. PGE2, a usual inflammation vector, and LTB4, TXB2, that are involved in the onset and deterioration of inflammation, were all significantly decreased. The levels of white blood cells, neutrophiles, and mononucleophiles also decreased, although the numbers were not significantly large. I*mmune-modulation of KKBT in the pathological mechanism of cartilage deterioration by inflammatory cells and their vectors was proved. This study should provide basis for the development of effective therapeutics as well as use in clinical practice.

Co-expression of Gamma-Aminobutyrate Aminotransferase and Succinic Semialdehyde Dehydrogenase Genes for the Enzymatic Analysis of Gamma-Aminobutyric Acid in Escherichia Coli

  • So, Jai-Hyun;Lim, Yu-Mi;Kim, Sang-Jun;Kim, Hyun-Ho;Rhee, In-Koo
    • Journal of Applied Biological Chemistry
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    • v.56 no.2
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    • pp.89-93
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    • 2013
  • Gamma-aminobutyric acid (GABA) aminotransferase (gabT) and succinic semialdehyde dehydrogenase (gabD) genes from Pseudomonas fluorescens KCCM 12537 were cloned into a single pETDuet-1 vector and co-expressed in Escherichia coli BL21(DE3) simultaneously. The mixture of both enzymes, called GABase, is the key enzyme for the enzymatic analysis of GABA. The molecular mass of the GABA aminotransferase and succinic semialdehyde dehydrogenase were determined to be 52.8 and 46.7 kDa following computations performed with the pI/Mw program, respectively. The GABase activity between pH 6.0 and 9.0 for 24 h at $4^{\circ}C$ remained over 75%, but under pH 6.0 decreased rapidly. The GABase activity between 25 and $35^{\circ}C$ by the treatment at pH 8.6 for 30 min remained over 80%, but over $35^{\circ}C$ decreased rapidly. When the activity against GABA was defined as 100%, the purified GABase activity against 5-aminovaleric acid having a similar structure to GABA showed 47.7% and GABase activity against ${\beta}$-alanine, ${\varepsilon}$-amino-n-caproic acid, $_L$-ornithine, $_L$-lysine, and $_L$-aspartic acid showed between 0.3 to 2.3%. The GABA content was analyzed with this co-expressed GABase, compared with the other GABase which was available commercially. As a result, the content of GABA extracted from brown rice, dark brown rice, and black rice were $26.4{\pm}3.5$, $40.5{\pm}4.7$ and $94.7{\pm}9.3{\mu}g/g$, which were similar data of other GABase in the error ranges.