• 제목/요약/키워드: Osmotic activity

검색결과 102건 처리시간 0.024초

고삼투압이 재조합 Erythropoietin의 생산과 당쇄구조에 미치는 효과

  • 정연태;김정회
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2001년도 추계학술발표대회
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    • pp.221-224
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    • 2001
  • Effect of hyperosmotic pressure on growth of recombinant Chinese hamster 。 vary cells and Erythropoietin (EPO) production was investigated. Cells were cultivated in batch modes at various osmolalities. When the osmolality increased from 314 to 463mOsm/Kg, specific EPO productivity (qp) was increased up to 1.6-fold but cell growth was inhibited. EPO has a complex oligosaccharide structure that plays an important role in biological activity in vivo. To investigate the influence of hypoerosmotic pressure on the glycosylation, structural analysis of oligosaccharide was calTied out. Recombinant human EPO was produced by CHO cells grown under various osmotic pressure and purified from culture supernatants by heparin-sepharose affinity column and immunoaffinity column. N-linked oligosaccharides were released enzymatically and isolated by paper chromatography. The isolated oligosaccharides were labeled with fluorescent dye, 2-aminobenzamide and analyzed with MonoQ anion exchange chromatography and GlycosepN amide chromatography for the assignment of GU (glucose unit) value. Glycan analysis by HPLC showed that neutral (asialo) oligosaccharide was increased slightly with an increase in osmolality. In portion of sialylated glycan, total relative amount of mono- and di-sialyated glycan was increased but that of tri- and tetra-sialylated glycan decreased as osmolality was increased.

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Fusarium poae와 Fusarium sporotrichioides간의 원형질체 융합 (Interspecific Protoplast Fusion between Fusarium poae and Fusarium sporotrichioides)

  • 하경란;장성렬;민병례
    • 미생물학회지
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    • 제29권2호
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    • pp.123-129
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    • 1991
  • In order to develop the protoplast fusion method of the strains of Fusarium, the interspecific protoplast fusion was attempted between Fusarium poae and F. sporotrichioides. Various auxotrophic mutants were isolated by the treatment of N-Methyl-N'-Nitro-N-Nitrosoguanidine. The optimal conditions for the formation and regeneration of protoplasts were examined and the characteristics of a fusant were studied. As a results, protoplasts were readily obtained from 18 hours cultured mycelia by the treatment of driselase for 3 hours and 0.6 M KCl as a best osmotic stabilizer at pH 6.0 for the formation of protoplast. Sucrose was the most suitable for the regeneration. Polyetylene glycol (M.W. 8,000) in $CaCl_{2}$-glycine solution was used to induce the protoplast fusion. The interspecific fusion frequency between protoplasts among the auxotrophic mutants of the two strains ranged from $2.7*10^{-2}$ to $5.7*10^{-3}$ . DNA content and cellulase activity were rather increased in the interspecific fusant. The lag phase of growth curve was slightly elongated in the fusant.

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Fecal Retention in Overactive Bladder (OAB) in Children: Perspective of a Pediatric Gastroenterologist

  • Jeong, Su Jin
    • Childhood Kidney Diseases
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    • 제19권1호
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    • pp.1-7
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    • 2015
  • Coexisting voiding and bowel dysfunction in children are common in the clinic. The idea that overactive bladder (OAB) and constipation arise from one single pathophysiology has been reinforced in many studies. In Korea, a nationwide multicenter study conducted in 2009 showed that overall prevalence of OAB in children, 5-13 years of age, was 16.59% and this number has increased more recently. The initial step to manage coexisting fecal retention and OAB in children is to characterize their bowel and bladder habits and to treat constipation if present. Although diagnosing constipation in children is difficult, careful history-taking using the Bristol Stool Form Scale, and a scoring system of plain abdominal radiography, can help to estimate fecal retention more easily and promptly. Non-pharmacological approaches to manage functional constipation include increasing fluids, fiber intake, and physical activity. Several osmotic laxatives are also effective in improving OAB symptoms and fecal retention. Additionally, correction and education in relation to toilet training is the most important measure in treating OAB with fecal retention.

Optimization of Parameters for GUS Gene Transformation of Porphyra yezoensis by Particle Bombardment

  • Nam, Bo-Hye;Park, Jung-Youn;Jin, Deuk-Hee;Hong, Yong-Ki
    • Fisheries and Aquatic Sciences
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    • 제9권4호
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    • pp.135-139
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    • 2006
  • We optimized the biological and physical parameters for DNA delivery into thalli of the red alga Porphyra yezoensis using a particle bombardment device. The efficiency of transformation was determined using the ${\beta}-glucuronidase$ (GUS) assay. The optimal helium pressure, distance of tungsten particle flight, and ratio of DNA to tungsten particles were $23kgf/cm^2$, 8 cm, and $5{\mu}g/mg$ tungsten, respectively. During bombardment, osmotic treatment with a mixture of 0.6 M mannitol and sorbitol increased the efficiency of GUS transformation. After 2 days, the blue color indicating GUS activity was observed using a histochemical assay.

재조합 Saccharomyces cerevisiae에서 Permeabilizing Agent를 이용한 HLY의 분비촉진 (Enhancement of Human Lysozyme Secretion with Permeabilizing Agents from the Recombinant Saccharomyces cerevisiae)

  • 황용일;최선욱;하정욱
    • 한국식품영양과학회지
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    • 제28권2호
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    • pp.348-352
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    • 1999
  • We have intended to accelerate the secretion of human lysozyme(HLY) with permeabilizing agents from the cultivated cells of the recombinant Saccharomyces cerevisiae. The five agents CaCl2, Tween 80, ethanol, Triton X 100, and cetyltrimethylammonium bromide(CTAB) were used as permeabilizing agents. Treatments of the yeast cell with CaCl2, Tween 80, and ethanol were effective to increase the secretion from the yeast cells. Especially, treatment of 10% ethanol increased the extracellular HLY activity by 38.6% at 30oC for 48 h in culture broth. But Triton X 100 and CTAB unexpectedly didn't play a role in increase of HLY secretion. Recovery of a foreign protein by permeabilizing agents is easier than by osmotic shock, and is less expensive than enzymatic digestion.

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Introduction and Expression of Foreign Genes in Rice Cells by Particle Bombardment

  • Jeon, Jong-Seong;Jung, Hou-Sung;Sung, Soon-Kee;Lee, Jong-Seob;Choi, Yang-Do;Kim, Han-Jip;Lee, Kwang-Woong
    • Journal of Plant Biology
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    • 제37권1호
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    • pp.27-36
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    • 1994
  • For establishing a transformation system of rice, an efficient introduction of foreign genes into embryogenic cell suspension by particle bombardment was conducted. The particle inflow gun based on the acceleration of DNA-coated tungsten particles using pressurized helium was constructed for delivery of DNA into rice cells. Several bombardment parameters were optimized using the transient expression of GUS gene. The conditions that gave the highest GUS gene expression of about 1000 blue spots per g fresh weight of bombarded cells include treatment of the cells with 0.5 M osmotic pressure, and use of the 410 kPa helium, 110 mm target distance, 13 mm syringe filter holder and 5 $\mu$L DNA/tungsten mixtures. It was also confirmed that rice actin promoter-intron construct gave the highest expression of all promoter-sequences studied. Eight weeks after the bombardment, stably transformed calluses were obtained on the selection medium containing 100 mg/L G418 and showed the strong activity in in situ GUS assay.

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흰쥐 대뇌피질에서 $A_1$ 아데노신 수용체의 탈감작 (Desensitization of $A_1$ Adenosine Receptors in Rat Cerebral Cortex)

  • 박경선;양완숙;김경환
    • 대한약리학회지
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    • 제32권2호
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    • pp.151-158
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    • 1996
  • Following the subcutaneous administration of $R(-)N^6-(2-phenylisopropyl)adenosine(600\;nmol/kg/hr)$ to rats for 1 week using t$Alzet^{\circledR}$ mini-osmotic pumps, $A_1$ adenosine receptor functions were determined using $[^3H]DPCPX$ binding, $[^{35}S]GTP_{\gamma}S$ binding, and adenylyl cyclase assays. $A_1$ adenosine receptor binding and the inhibition of adenylyl cyclase activity by PIA was not altered in cerebrocortical membranes prepared from PIA-treated rats. However, there was a significant decrease in the $A_1$ adenosine receptor-mediated stimulation of $[^{35}S]GTP_{\gamma}S$ binding to cerebrocortical membranes prepared from PIA-treated rats(22.0% decrease in basal activity; 19.7% decrease in maximal activity). These results suggest that the desensitization of $A_1$ adenosine receptors following chronic administration involves agonist-induced uncoupling of the receptors from G proteins rather than alteration of $A_1$ adenosine receptor molecules. It is also suggested that the determination of stimulation of $[^{35}S]GTP_{\gamma}S$ binding to G proteins is a suitable tool in studying the receptor regulation including desensitization

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Changes in Acrosin Activity and Membrane Function of Boar Spermatozoa

  • Kim, C.K.;Kim, I.;Y.C. Chung;J.W. Ryu;H.J. Yoon;K. Kang;Kim, I.C.;Lee, J.H.;S.E. Yeon
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2001년도 춘계학술발표대회
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    • pp.86-86
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    • 2001
  • The aims of this work were to determine the acrosin activity and to evaluate the structural and functional integrity of AS boar spermatozoa. The acrosin activity of spermatozoa were 5.40, 4.10 and 3.40 mIU/10$^{6}$ sperm in raw, extended and frozen semen respectively , which differed significantly each other (P<0.05). After the raw and extended semen were exposured to cold and thermal shock, the acrosin activities of spermatozoa in the raw semen were 5.39, 5.21 and 5.29 mIU/10$^{6}$ sperm for control (non-shock), cold shock and thermal shock, and those of extended semen were 4.21, 3.98 and 4.00 mIU/10$^{6}$ sperm. This value among treatments did not differ significantly. The acrosin activities of spermatozoa in the extended and stored semen were 3.27, 3.52, 3.46 and 3.23 mIU/10/suup 6/ sperm, while hypo-osmotic test(HOST) values were 56.5%, 64.7%, 66.0% and 56.0%, following 4 days storage at 4$^{\circ}C$, 17$^{\circ}C$ , $25^{\circ}C$ and 37$^{\circ}C$, respectively. The results at 17$^{\circ}C$ and $25^{\circ}C$ appeared to be best compared with the other storage temperatures.

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Enhanced drought and salinity tolerance in transgenic potato plants with a BADH gene from spinach

  • Zhang, Ning;Si, Huai-Jun;Wen, Gang;Du, Hong-Hui;Liu, Bai-Lin;Wang, Di
    • Plant Biotechnology Reports
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    • 제5권1호
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    • pp.71-77
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    • 2011
  • Drought and salinity are the most important abiotic stresses that affect the normal growth and development of plants. Glycine betaine is one of the most important osmolytes present in higher plants that enable them to cope with environmental stresses through osmotic adjustment. In this study, a betaine aldehyde dehydrogenase (BADH) gene from spinach under the control of the stress-induced promoter rd29A from Arabidopsis thaliana was introduced into potato cultivar Gannongshu 2 by the Agrobacterium tumefaciens system. Putative transgenic plants were confirmed by Southern blot analysis. Northern hybridization analysis demonstrated that expression of BADH gene was induced by drought and NaCl stress in the transgenic potato plants. The BADH activity in the transgenic potato plants was between 10.8 and 11.7 U. There was a negative relationship (y = -2.2083x + 43.329, r = 0.9495) between BADH activity and the relative electrical conductivity of the transgenic potato plant leaves. Plant height increased by 0.4-0.9 cm and fresh weight per plant increased by 17-29% for the transgenic potato plants under NaCl and polyethylene glycol stresses compared with the control potato plants. These results indicated that the ability of transgenic plants to tolerate drought and salt was increased when their BADH activity was increased.

주요작물의 한발저항성에 관한 연구 제2보 맥류 유묘기의 수분부족이 질산환원효소 및 단백질 분해효소의 활성변화와 유리 Proline의 축적에 미치는 영향 (Studies on the Drought-Resistance of Major Food Crops II. Effect of Water Stress on the Activity of Nitrate Reductase and Protease, and the Accumulation of Free Proline in Barley and Wheat at Seedling Stage)

  • 최원열;민경수
    • 한국작물학회지
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    • 제27권1호
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    • pp.60-65
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    • 1982
  • 맥류의 한발저항성 반응과 정도 그리고 저항생리적 기작을 구명하기 위하여 발아후 10일된 유묘기(본엽 3매시)에 8일간 단수처리를 하여 질산환원효소와 단백질분해효소의 활성변화 그리고 proline축적을 조사하였던 바 그 결과는 다음과 같다. 1. 전품종의 평균 감소율은 질산환원효소의 활성이 42%이였으며 단백질분해효소의 활성은 73%였으며, 이에 반하여 proline은 대조구에 비하여 단수구가 무려 10배나 증가하였다. 2. 질산환원효소의 활성감소율을 맥종별로 보면 소맥 < 호맥 < 대맥 < 2조대맥ㆍ과맥의 순으로 소맥이 가장 낮았다. 3. 단백질분해효소의 활성감소율을 맥종별로 보면 소맥 < 호맥 < 이조대맥 < 대맥 < 과맥의 순으로 소맥이 높고 과맥이 가장 낮았다. 4. 단수구의 proline의 축적색대량을 맥종별로 보면 소맥 < 대맥 < 호맥 < 과맥 < 이조대맥의 순으로 소맥과 대맥이 높았으며, 대조구에 대한 단수구의 증가비는 호맥(13배) > 소맥ㆍ대맥(11배) > 과맥(9배) > 이조대맥(7배)의 순으로 호맥이 가장 높았다. 5. 한발저항성 과정에서 효소적 및 생리적 대사작용의 관점에서 볼 때 소맥 > 호맥 > 대맥 > 과맥 > 이조대맥의 순으로 한발저항성이 강한 것으로 추정된다.

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