• 제목/요약/키워드: HEPES

검색결과 113건 처리시간 0.024초

A New Rhodamine B Hydrazide Hydrazone Derivative for Colorimetric and Fluorescent "Off-On" Recognition of Copper(II) in Aqueous Media

  • Tang, Lijun;Guo, Jiaojiao;Wang, Nannan
    • Bulletin of the Korean Chemical Society
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    • 제34권1호
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    • pp.159-163
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    • 2013
  • A new Rhodamine B hydrazide hydrazone 1 has been synthesized and investigated as a colorimetric and fluorescent "off-on" sensor for the recognition of $Cu^{2+}$ in $CH_3CN/H_2O$ (1:1, v/v, HEPES 10 mM, pH = 7.0) solution. Sensor 1 displayed highly selective, sensitive and rapid recognition behavior toward $Cu^{2+}$ among a range of biologically and environmentally important metal ions. Sensor 1 bind $Cu^{2+}$ via a 1:1 stoichiometry with an association constant of $1.92{\times}10^6\;M^{-1}$, and the detection limit is evaluated to be $7.96{\times}10^{-8}\;M$. The $Cu^{2+}$ recognition event is reversible and is barely interfered by other coexisting metal ions.

진행된 암 동물모델에서의 리포좀 포집 PALA의 항암 치료 효과 (Therapeutic Potency of N-(Phosphonacetyl)-L-Aspartic Acid in Liposome in Established Tumor Bearing Mice)

  • 김진석
    • Journal of Pharmaceutical Investigation
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    • 제30권2호
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    • pp.127-131
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    • 2000
  • Previously, we have reported an antitumor efficacy of liposomal N-(phosphon-acetyl)-L-aspartic acid (or PALA) in C-26 tumor bearing Balb/c mice, where PALA in liposome was administered one day after tumor inoculation. In this report, we have investigated the therapeutic potency of liposomal formulation of PALA, which was administered eight days after tumor inoculation in the same C-26 tumor bearing mice. The C-26 murine colon tumor inoculated mice were randomized for the in vivo therapy and the survival was measured after a single intraperitoneal injection of the drug. When the therapy was initiated eight days after tumor inoculation, DSPC-PALA at 150 mg/kg resulted in a significant increase in median survival time (MST) of 56% over the control group which received MES/HEPES buffer alone. However, none of the free PALA and DSPG-PALA liposome doses caused a statistically significant increase in MST over control group at the 95% confidence level. At 750 mg/kg dose, free PALA caused a marginally significant improvement in MST by 34%, but both 375 mg/kg and 150 mg/kg doses of free PALA caused only a 2% and a 4% increase in MST, respectively. These results show that PALA in neutrally charged liposome can exhibit considerably greater potency than free PALA in established C-26 tumor bearing mice.

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우심낭을 이용한 이종이식 보철편의 개발 (II) - 0.625% Glutaraldehyde 에 보존한 우심낭의 임상 적용 - (Investigation of Bovine Pericardial Heterograft (II0) : Clinical applications of glutaraldehyde-preserved bovine pericardium)

  • 김기봉
    • Journal of Chest Surgery
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    • 제23권3호
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    • pp.465-473
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    • 1990
  • Glutaraldehyde have been used as the most effective cross-linking agent for stabilizing collagen fibers and preventing biodegradation. We processed bovine pericardium in a solution containing 0.625% glutaraldehyde,0.05M HEPES buffer and 0.26% magnesium chloride in saline. The glutaraldehyde-preserved bovine pericardium was implanted in 36 patients at Seoul National University Hospital during a 11-month period between May 1989 and March 1990. 24 were males and 12 females, with ages ranging from 6 months to 168 months [mean age of 43 months]. In 12 patients, the glutaraldehyde-preserved bovine pericardium was used for orthotopic reconstruction of the pericardial sac. In 24 patients. the glutaraldehyde-preserved bovine pericardium was heterotopically implanted.; pulmonary monocusp implant and RVQT [right ventricular outflow tract] patch widening were performed in 10 patients, pulmonary monocusp implant in 6, RVOT patch widening in 4, valved conduit in 2, conduit and pulmonary angioplasty in 1, and ventricular septation in l. With vascular suture techniques, the anastomoses were immediately tight. There was no bleeding from the needle holes and no oozing through bovine pericardium itself. During the follow-up period of up to 10 months, no infections of the glutaraldehyde-preserved bovine pericardium occurred and no bovine pericardium-related complications were observed in this series.

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인삼(人蔘) Saponin이 Prolactin 분비(分泌)에 미치는 효과(效果) (The Effects of Ginseng Saponin on Prolactin Secretion in Rats)

  • 백덕우;이서윤;지형준
    • 생약학회지
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    • 제9권1호
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    • pp.33-39
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    • 1978
  • The present study is involved with the prolactin secretion from anterior pituitary gland by ginseng saponin since it was handled down by tradition that ginseng might influence the milk secretion when it was given to nursing mother. To investigate the effect of saponin on the prolactin production or release from the anterior pituitary gland, cell culture study and whole animal studies were carried out. For the cell culture study, enzymatically dispersed anterior pituitary cells of rat anterior pituitary gland in HEPES buffers containing trypsin were used. Ginseng saponin was added to the culture media and the amount of prolactin produced in the cell culture media was determined by radloimmunoassay(RIA) technique. Dose-dependent increases of prolactin with ginseng saponin were observed, whereas, no change was observed without ginseng treatment. For the whole animal study, normal and castrated rats which previously cannulated into the heart via the right juglar vein were used. The prolactin concentration in plasma were determined by using the technique of RIA. In normal rats, prolactin concentration in plasma were elevated dramatically after 1 hour of ginseng saponin administration, whereas, instantaneous increases were observed in castrated rats. For prolactin assay by RIA, NIAMDD Rat Prolactin Kit and NIAMDD Rat Prolactin RP-1 were used as standard. The results indicate that ginseng saponins increase the release of prolactin from the anterior pituitary gland and production of prolactin from the cell in rats.

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Comparison of Developmental Efficiency of Murine Somatic Cell Nuclear Transfer Protocol

  • Moon, Jeonghyeon;Jung, Miran;Roh, Sangho
    • 한국수정란이식학회지
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    • 제32권3호
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    • pp.81-86
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    • 2017
  • The Somatic cell nuclear transfer (SCNT) method can be applied to various fields such as species conservation, regenerative medicine, farming industries and drug production. However, the efficiency using SCNT is very low for many reasons. One of the troubles of SCNT is that it is highly dependent on the researcher's competence. For that reason, four somatic cell nuclear injection methods were compared to evaluate the effect of hole-sealing process and existence of cytochalasin B (CB) on efficiency of murine SCNT protocol. As a results, the microinjection with the hole-sealing process, the oocyte plasma membrane is inhaled with injection pipette, in HCZB with CB was presented to be the most efficient for the reconstructed in SCNT process. In addition, we demonstrated that the oocytes manipulated in Hepes-CZB medium (HCZB) with CB does not affect the developmental rate and the morphology of the blastocyst during the pre-implantation stage. For this reason, we suggest the microinjection involving hole-sealing in HCZB with CB could improve SCNT process efficiency.

Crystallization and X-ray crystallographic analysis of the PH-like domain of lipid transfer protein anchored at membrane contact sites from Saccharomyces cerevisiae

  • Tong, Junsen;Im, Young Jun
    • Biodesign
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    • 제5권4호
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    • pp.136-140
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    • 2017
  • Lam6 is a member of sterol-specific ${\underline{l}ipid$ transfer proteins ${\underline{a}}nchored$ at ${\underline{m}ebrane$ contact sites (LAMs). Lam6 localizes to the ER-mitochondria contact sites by its PH-like domain and the C-terminal transmembrane helix. Here, we purified and crystallized the Lam6 PH-like domain from Saccharomyces cerevisiae. To aid crystallization of the Lam6 PH-like domain, T4 lysozyme was fused to the N-terminus of the Lam6 PH-like domain with a short dipeptide linker, GlySer. The fusion protein was crystallized under the condition of 0.1 M HEPES-HCl pH 7.0, 10% (w/v) PEG 8000, and 0.1 M $Na_3$ Citrate at 293K. X-ray diffraction data of the crystals were collected to $2.4{\AA}$ resolution using synchrotron radiation. The crystals belong to the orthorhombic space group $P2_12_12_1$ with unit cell parameters $a=59.5{\AA}$, $b=60.1{\AA}$, and $c=105.6{\AA}$. The asymmetric unit contains one T4L-Lam6 molecule with a solvent content of 58.7%. The initial attempt to solve the structure by molecular replacement using the T4 lysozyme structure was successful.

Effect of Cycloheximide on Bovine Oocyte Nuclear Progression and Sperm Head Transformation after Fertilization In Vitro

  • Liu, L.;Zhang, H.W.;Qian, J.F.;Fujihara, N.
    • Asian-Australasian Journal of Animal Sciences
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    • 제12권1호
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    • pp.22-27
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    • 1999
  • Bovine oocytes with compact and complete cumulus cells were cultured in 6 groups for up to 24h in TCM199 buffered with 25 mmol/1 HEPES and supplemented with 10% FCS, 1 mg/ml $17{\beta}$-estradiol, 20 IU/ml hCG. Half of the oocytes at each group cultured in the presence of $25{\mu}g/ml$ cycloheximide at different times during maturation (0, 6, 12, 18, 20, 22 h) were fixed at 24 h of maturation to examine the nuclear progression. The rests of them were inseminated with frozen-thawed spermatozoa in medium BO with 10 mg/ml BSA and 10 mg/ml heparin and fixed after additional 18-20 h culture to evaluate the sperm head transformation. When a protein synthesis inhibitor was added at the onset of the maturation, the oocytes were prevented to proceed GVBD. A few of the oocytes (16%) were able to be penetrated and sperm head decondensation was inhibited either. Addition of cycloheximide after 6-12 h of culture resulted in an increasing percentage of GVBCD (more than 80%), but the oocytes became arrested in M-I (69.2%). More than half of the oocytes was penetrated with a decondensing sperm head. Formation of male pronucleus was first obtained at 12 h of culture in the presence of cycloheximide. When cycloheximide was added from 18 h of culture onwards, nuclear progression to M-II was increasingly restored (80.4-85.5%). The proportion of male and female pronuclear formation increased from 17.9% to 46.2%. It is concluded that protein synthesis is necessary not only for GVBD and development from M-I to M-II, but also for sperm head decendensation and male pronuclear formation in bovine oocytes.

Effects of Cryoprotectant, Warming Solution and Removal of Lipid on Viability of Porcine Nuclear Transfer Embryos Vitrified by Open Pulled Straw Method

  • Cong, Pei-Qing;Song, Eun-Sook;Kim, Eui-Sook;Li, Zhao-Hua;Zhang, Yong-Hua;Lee, Jang-Mi;Yi, Young-Joo;Park, Chang-Sik
    • Reproductive and Developmental Biology
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    • 제31권2호
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    • pp.103-108
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    • 2007
  • This study was carried out to investigate the effects of cryoprotectants, warming solution and removal of lipid on open pulled straw vitrification (OPS) method of porcine embryos produced by nuclear transfer (NT) of fetal fibroblasts. All solutions used during vitrification were prepared with holding medium consisting of 25 mM Hepes buffered TCM199 medium containing 20% fetal bovine serum (FBS) at $38.5^{\circ}C$. The blastocysts derived from NT with or without lipid were vitrified in each medium of different concentrations of dimethyl sulfoxide (DMSO) and ethylene glycol (EG). Also, blastocysts after cryopreservation were warmed into different concentrations of sucrose in warming solution. The optimal concentrations of cryoprotectants in vitrification solution were 10% DMSO + 10% EG in vitrification solution 1 (VS1) and 20% DMSO + 20% EG in vitrification solution 2 (VS2). The optimal concentrations of sucrose were 0.3 M sucrose in warming solution 1 (WS1) and 0.15 M sucrose in warming solution 2 (WS2). lipid removal from oocytes before NT enhanced the viability of NT embryos after vitrification. Our results show that use of the OPS method in conjunction with lipid removal provides effective cryopreservation of porcine nuclear transfer embryos.

Lactobacillus casei와 Lactobacillus delbrueckii간의 Protoplast 융합에 관한 연구 (Studies on the Protoplast Fusion between Lactobacillus casei and Lactobacillus delbrueckii)

  • 전홍기;김미경;백형석
    • 한국미생물·생명공학회지
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    • 제20권1호
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    • pp.6-13
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    • 1992
  • 유산균주의 균주개량방법의 일환으로 protoplast fusion 방법을 사용하여 lincomycin에 내성을 나타내는 Lactobacillus casei KCTC 1121과 rifampicin에 내성을 나타내는 Lactobacillus delbruckii JK-414의 protoplast 형성과 재생, 융합에 대한 조건 및 융합주의 생리학적 성질 등을 검토하였다. Lactobacillus case와 Lactobacillus delbrueckii JK-414는 삼투압 안정제로 sucrose가 함유된 protoplast forming buffer에서 5$\mu g$/ml의 mutanolysin 으로 $42^{\circ}C$, 15분간 처리 했을 때 protoplast 형성율이 높게 나타났다.

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평지방막에 융합된 관상동맥의 $Ca^{2+}$ - activated $K^{+}$ 통로의 특성

  • 김희정;박진봉;류판동;이문한;임병용;홍기환
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 1994년도 춘계학술대회 and 제3회 신약개발 연구발표회
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    • pp.185-185
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    • 1994
  • 관상동맥은 전기적으로 자극하여도 활동전위가 쉽게 유발되지 않는 조직인데, 이는 세포내의 $Ca^{2+}$에 의해서 활성화되는 $K^{+}$통로때문이라고 여겨지고 있다. 저자들은 단일 이온통로 수준에서 이러한 관상동맥 $K^{+}$ 통로의 성질을 조사하고, 아울러 $K^{+}$ 통로 개방제를 포함한 몇가지 약물이 $K^{+}$ 통로에 미치는 영향을 알아보고자 본 연구를 수행하였다. 막소포는 돼지의 심장에서 관상동맥을 분리하여 균질화 및 원심분리하여 준비하였으며, sucrose 20/30%에 있는 층을 취하여 분주한 후 -7$0^{\circ}C$로 보관하여 사용하였다. 평지방막은 1-palmitoyl, 2-oleoyl-phosphatidyl ethanolamine과 1-palmitoyl, 2-oleoyl phosphatidylcholine (4:1)을 n-decane에 녹인지방액(25mg/m1)을 200 $\mu\textrm{m}$의 구멍에 칠하여 만들었다. 단일 $K^{+}$ 통로는 막소포 (1-5$\mu\textrm{g}$/ml)를 평지방막의한쪽에 추가하고 자석으로 저어 융합이 되도록 하였으며, 통로의 활성은 막의 양쪽에 150 mM KCI, 10 mM HEPES-KOH, 0.5 mM EGTA, 0.6 mM $CaCl_2$ (pH 7.2)로 된용액이 존재하는 상태에서 막전압을 다르게 하며 기록하였다. 단일 이온 통로 전류는 amplifier를 통하여 VCR tape에 녹화하여 후에 분석에 이용하였으며, 동시에 종이기록지에 기록하였다. 단일이온통로 활성의 분석은 pClamp (Ver5.5)를 이용하여 수행하였다. 관상동맥에서 가장 흔히 기록되는 이온통로는 막전압과 $Ca^{2+}$의 농도에 의하여 변하는 $K^{+}$ 통로이었는데 pCa 4.0과 -10 mV에서 초기 활성도(Po)는 0.1에서 0.8까지 다양하였다. 전류와 전압과의 관계는 -60 - +60mV의 전압범위에서 직선형이었다.

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