• Title/Summary/Keyword: Da No.

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Internal Cleavage of Bacillus subtilis BSE616 Endo-$\beta$-1, 4-glucanase expressed in Escherichia coli

  • KIM, HOON;SUNGMIN F. KIM;DONG HO AHN;JlN HO LEE;MOO YOUNG PACK
    • Journal of Microbiology and Biotechnology
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    • v.5 no.1
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    • pp.26-30
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    • 1995
  • The cytoplasmic endo-$\beta$-l, 4-glucanase (endoglucanase) was purified from cell extracts of Escherichia coli (pBS1) transformant carrying the Bacillus subtilis endo-$\beta$-l, 4-glucanase gene after full growth, and its molecular weight was found to be 52 kilodaltons (kDa). The endo-$\beta$-l, 4-glucanase isolated from the periplasmic space was smaller than 52-kDa cytoplasmic enzyme. The 52-kDa endoglucanase was found to be cleaved in the periplasm and finally converted to 34.5-kDa protein. Small amounts of both 52-kDa and 34.5-kDa proteins were secreted into the culture broth. The cleavage took place in the C-terminal portion of the enzyme. The N-terminal amino acid residues of both 52-kDa and 34.5-kDa enzymes were determined to be the same, Ala, the 30th residue of the primary translation product. Cleavage of the C-terminal portion showed to have no significant effect on the basic enzyme properties.

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A Four-Week Intravenous Toxicity Study of DA-l131/betamipron in Rats (DA-l131/betamipron의 랫드에 대한 4주반복 정맥투여 독성시험)

  • Kim, Dong-Hwan;Cho, Hyeon;Kang, Kyung-Koo;Baik, Nam-Gi;Kim, Won-Bae
    • Biomolecules & Therapeutics
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    • v.6 no.2
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    • pp.204-212
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    • 1998
  • This study was conducted to evaluate the repeated dose toxicity of DA-1131/betamipron, newly developed carbapenem antibiotic, in rats. DA-1131/ betamipron was administered intravenously once a day for 4 weeks to 10 males and 10 females per group at the doses of 0(control),40, 160 and 640 mg/kg. Throughout the study period, all rats survived. The administration of DA-1131/betamipron induced soft stool or diarrhea in rats of both sexes receiving 160 or 640 mg/kg. The water consumption was increased with a statistical significance in 640 mg/kg during observation period. At the end of administration, hematological and serum biochemical examination showed no toxicological changes in DA-1131/betamipron treated groups compared with control group. Histopathologic examination revealed inflammatory cell infiltration, tubular dilatation and focal necrosis of kidney in two males and three females in 640 mg/kg. On the basis of these results, the noobserved-adverse-effect-level of DA-1131/betamipron was estimated to be 40 mgtg under the present test condition.

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Distribution of Phyrase in The Development Rat Organs (성장 발육에 따른 흰쥐 장기내 phytase의 분포성)

  • 양원진
    • Journal of Life Science
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    • v.7 no.2
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    • pp.127-133
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    • 1997
  • The phytase(myo-inositol hexkisphosphate phosphohydrolase ; EC 3.1.3.8) activity was observed only in the homogenate of intestinal mucosa, though the activity of alkaline phisphatase was measurable in various organs. In addition, no protein bands were detected in any other organs on immunoblotting using the anti-90kDa phytase antiserum. Thses results suggest that phytase is specifically present in small intestinal mucosa, and that hydrolysis of phytic acid(inositol-hexakisphosphate) can be allotted for a physiological role of the intestine-specific enzyme. The activities of phytase was increased during development of rat. The 70kDa phytase appeared just after birth, but the 90kDa phytase was not observed until adult period, suggesting that the 90kDa phytase was synthesized in response to weanling.

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Changes in Pectoral Mvoblast Proteins- during Myofibrillogenesis in vitro (배양흉근 근모세포의 근원섬유 형성과정 동안의 근단백질의 양상)

  • 하재청;김한도김병기
    • The Korean Journal of Zoology
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    • v.35 no.3
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    • pp.322-331
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    • 1992
  • To investigate the svnthyesis of muscle proteins during differentiation of chicken myoblast, cvtosolic and membrane fractions were used for both sodium dodecvl sulfate polvcrylamide gel eBectrophoresis and two-dimensional gel electrophoresis. An extensive cell fusion was observed in 4 day culture. In the protein pattern of the cvtosolic fraction from SDS-PAGE. several protein bands including 250 kDa and 46 kDa showed remarkable changes during culture. the protein of 46 kDa was the most prominent one ann its optical density was the highest in 5 day culture (OD = 1.30). In the membrane fraction, band of 19.8 kDa showed the highest absorbance with 0.93 OD at 12 hr after initial plating and decreased gradually thereafter to 0.23 in 5 nay culture. From the results of two-dimensional gel electrophoresis of cytosolic fraction, the 46 kDa spot was observed as ko separated forms from culture 2 nary culture, and the sixte of this spot was the largest in 5 nay culture. In the pattern of membrane protein, the extensive appearance of newiv synthesized Proteins was found in a naut culture, but no Prominent spot was observed throughout culture. From the results of the present clay, we found that, during myoblast differentiation, the most prominent proteins were bands of 46 kDa and 19.8 kDa in cvtosolic and membrane fraction, respectively, and the appearance of new proteins was initiated at 48 hr after initial plating, and the 46 kDa protein was predominant in the cytoplasm of late culture in which extensive cell fusion was observed.

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Studies on The Molecular Mechanism of 33 kDa extrinsic Protein in Photosystem II Oxygen-Evolving Complex

  • Xu, Chunhe;Ruan, Kangcheng;Yu, Yong;Weng, Jun
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.82-85
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    • 2002
  • 33kDa extrinsic protein, an important protein in oxygenic photosynthesis, was known to have no fixed configuration in solution. At 20$\^{C}$ and pH 6, 33kDa extrinsic protein showed changes of free energy of -14.6 kJ/mor$\^$-1/ and of standard volume of -120mL/mol, respectively, with increase of hydrostatic pressure, comparatively lower than for most proteins. NBS modification of Trp241 in 33kDa extrinsic protein dramatically changes the secondary protein structure, its affinity to photosystem II as well as photosynthetic oxygen evolution. The relationship between structural change and transport of oxygen, water and proton is deserved a further study.

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Reproductive Toxicity of DA-125, A New Anthracycline Anticancer Agent: Teratogenicity Study in Rabbits (새로운 안트라사이클린계 항암제 DA-125의 생식독성연구: 토끼 최기형시험)

  • 정문구;김종춘;한상섭;노정구
    • Biomolecules & Therapeutics
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    • v.3 no.1
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    • pp.47-53
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    • 1995
  • DA-125, a new anthracycline antitumor antibiotic, was administered at dose levels of 0, 0.2, 0.6 and 1.8 mg/kg/day intravenously to pregnant New Zealand White rabbits from day 6 through 18 of gestation. The does were subjected to the caesarean section on day 28 of gestation. Effects of test agent on general toxicity of does and embryonic development of F1 fetuses were examined. At 1.8 mg/kg, the organ weight for ovary of does was significantly decreased. The decrease in the number of corpus lutea, implantations and litter size, and the increase in the rate of resorptions were also observed. In addition, various types of external, visceral and skeletal malformations occurred in fetuses at an incidence of 7.7, 7.7 and 20.6%, respectively. The results show that the no effect dose levels (NOELs) of DA-125 are 0.6 mg/kg/day for does and F1 fetuses.

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Pichia pastoris: A Recombinant Microfactory for Antibodies and Human Membrane Proteins

  • Goncalves, A.M.;Pedro, A.Q.;Maia, C.;Sousa, F.;Queiroz, J.A.;Passarinha, L.A.
    • Journal of Microbiology and Biotechnology
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    • v.23 no.5
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    • pp.587-601
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    • 2013
  • During the last few decades, it has become evident that the compatibility of the yeast biochemical environment with the ability to process and translate the RNA transcript, along with its capacity to modify a translated protein, are relevant requirements for selecting this host cell for protein expression in several pharmaceutical and clinical applications. In particular, Pichia pastoris is used as an industrial host for recombinant protein and metabolite production, showing a powerful capacity to meet required biomolecular target production levels in high-throughput assays for functional genomics and drug screening. In addition, there is a great advantage to using P. pastoris for protein secretion, even at high molecular weights, since the recovery and purification steps are simplified owing to relatively low levels of endogenous proteins in the extracellular medium. Clearly, no single microexpression system can provide all of the desired properties for human protein production. Moreover, chemical and physical bioprocess parameters, including culture medium formulation, temperature, pH, agitation, aeration rates, induction, and feeding strategies, can highly influence product yield and quality. In order to benefit from the currently available wide range of biosynthesis strategies using P. pastoris, this mini review focuses on the developments and technological fermentation achievements, providing both a comparative and an overall integration analysis. The main aim is to highlight the relevance and versatility of the P. pastoris biosystem to the design of more cost-effective microfactories to meet the increasing demands for recombinant membrane proteins and clinical antibodies for several therapeutic applications.

Identification of Allergens of Dermatophagoides farinae on Canine Atopic Dermatitis in Korea (국내 개에서 발생한 아토피성 피부염에서 Dermatophagoides farinae의 항원 확인)

  • Cho, Na-Young;Park, Seong-Jun
    • Journal of Veterinary Clinics
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    • v.31 no.6
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    • pp.490-494
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    • 2014
  • Dermatophagoides farinae plays important role in the pathogenesis of canine atopic dermatitis as environmental allergens. Also, many studies revealed that D. farinae was the main causative allergen for Korean dogs with atopic dermatitis. To identify major allergens of D. farinae in Korean atopic dogs allergic to D. farinae by immunoblot using commercial allergenic extracts, 26 dogs from two groups were enrolled in the study. Control group consists of 10 dogs with no clinical signs of disease and atopic group consists of 16 dogs diagnosed as atopic dermatitis. Sera from Korean dogs with atopic dermatitis showed six allergens of D. farinae extract by procedure of immunoblot. The molecular weights of identifying protein bands were 177, 109, 75, 44, 27, 15 kDa. The major allergens showing reactivity with greater than 50% of atopic dogs were detected at approximately 44, 109 and 177 kDa. Subsequent investigations will be carried out to verify the identity of the allergens detected in this study.

Permeability of a Capsaicin Derivative $[{14}^C]DA-5018$ to Blood-Brain Barrier Corrected with HPLC Method

  • Kang, Young-Sook;Kim, Jong-Mi
    • Archives of Pharmacal Research
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    • v.22 no.2
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    • pp.165-172
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    • 1999
  • In the present work , the transport mechanism of a capsaicin derivative, DA-5018, through blood-brain barrier (BBB) has been investigated to evaluate the feasibility of potential drug development. The result of pharmacokinetic parameters obtained from the intravenous injection of plasma volume marker,$[3^H]RSA$ and $[{14}^C]DA-5018$, indicated that both AUC, area under the plasma concentration curve and VD, volume of distribution in brain of $[3^H]RSA$ agreed with those reported ($1620{\pm}10 $percentage injected dose minute per milliliter (%IDmin/ml) and $12.0{\pm}0.1{\mu}l/g$, respectively). Elimination half-life and AUC of $[{14}^C]DA-5018$is corrected by the PHLC analysis, 19.6$\pm$1.2 min and 7.69$\pm$0.85% IDmin/ml, respectively. The metabolic rate of $[{14}^C]DA-5018$was very rapid. The blood-brain barrier permeability surface area (PS) product of $[{14}^C]DA-5018$ was calculated to be 0.24$\pm$0.05 $\mu$l/min/g. The result of internal carotid artery perfusion and capillary depletion suggested that [14C]DA-5018 pass through BBB with the time increasingly. Investigation of transport mechanism of $[{14}^C]DA-5018$ using agonist and antagonist suggested that vanilloid (capsaicin) receptor did not exist in the BBB, and nutrient carrier system in the BBB has no effect on the transport of DA-5018. In conclusion, despite the fact that penetration of DA-5018 through BBB is significant, the intact drug found in the brain tissue is small because of a rapid metabolism. Therefore, for the central analgesic effect of DA-5018, the method to increase the metabolic stability in plasma and the brain permeability should be considered.

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Plasma concentration of dopamine varies depending on breed, sex, and the genotype of DRD4 in horses

  • Junyoung, Kim;Heejun, Jung;Jae-Young, Choi;Jang-Won, Lee;Minjung, Yoon
    • Journal of Animal Science and Technology
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    • v.64 no.4
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    • pp.792-799
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    • 2022
  • Dopamine (DA) is known to be a key modulator of animal behaviors. Thus, the plasma concentration of DA might be used as a biomarker for the behavioral characteristics of horses. The behavioral characteristics of horses vary depending on the breed, age, and sex. Moreover, the DA receptor genotypes are also related to horse behaviors. Thus, the aim of this study was to investigate the DA concentration variations of horse plasma by breed, age, sex, or genotype of its receptor. The horses were divided by breed into Thoroughbred (n = 13), Pony (n = 9), Warmblood (n = 4), and Haflinger (n = 5). The age variable was divided into three different groups: post-pubertal (2-5 years, n = 6), adult (6-13 years, n = 19), and aged horses (15-24 years, n = 6). The sex variable was divided into geldings (n = 8) and mares (n = 23). Approximately 10 mL of blood was collected, and an ELISA kit was used to measure the plasma concentration of DA. Polymerase chain reaction analysis was performed to identify the genetic variation in the DA D4 receptor gene (DRD4). SPSS statistical software was used for statistical analysis. The DA concentrations in geldings were significantly lower than those in mares. There was no significant difference in DA concentrations among breed and age groups. Horses with the GG and GA genotypes had significantly higher plasma concentrations of DA compared to horses with the AA genotype for the G292A gene. Briefly, the plasma concentration of DA varied depending on the sex and genotype of G292A. These factors should be considered when the concentration of DA is used as a biomarker for the behavioral characteristics of horses. In conclusion, the DA concentration or DRD4 genotype of horse plasma has the potential to be used as a biomarker that can predict the behavioral characteristics of horses.