• Title/Summary/Keyword: autoradiography

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뇌로의 약물송달(2) -혈액-뇌관문(BBB)의 Osmotic Opening법에 의한 뇌로의 약물송달

  • 황만용;박경호;손영택;이민화
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1993.04a
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    • pp.177-177
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    • 1993
  • 뇌모세혈관내피세포로 구성된 혈액-뇌관문(BBB)은 tight junction으로 구성되어 있을뿐만 아니라 fenestra가 없는 등 BBB가 가진 특성으로 인해 물질 수송에 장벽역할을 하기 때문에 주로 중추신경계 질환시의 약물요법에 많은 제한이 따른다. 따라서 뇌로의 효율적인 약물 송달방법에 대해 많은 연구가 진행중에 있는데 본 연구에서는 BBB의 hyperosmotic opening법을 이용하여 뇌로 잘 수송되지 않는 약물의 뇌송달 방법에 대해 연구하고자 하였다. 먼저 수용성으로 인해 BBB 에 잘 통과하지 않는 물질로 신장 및 뇌영상 촬영시 사용되는 방사성 의약품이 Tc-DTPA를 이용하여, hyperosmotic opening법에 의한 뇌투과 증가정도를 검토하였으며, 실험방법이 확립됨에 따라 뇌암의 항암요법을 위해 뇌로의 투과가 적은 항암제(5-Fluorouracil, 5-FU)의 뇌송달에 대해 본 방법을 적용하여 검토하였다. 실험동물로는 S.D.계 웅성 랫트를 사용하였으며, 실험방법은 다음과 같이 하였다. 랫트의 좌측 외경동맥(left external carotid artery)에 혈류의 역방향으로 혈관 분지점에서 1-2mm 전까지 PE-50 catheter를 삽입하고, 1.6 molal L-(+)- arabinose 고장액(1580mOsm)을 0.12ml/sec의 일정한 속도로 30초간 infusion함으로 BBB를 opening 한 다음, 5분 후에 Tc-DTPA를 대퇴 정맥으로 주사하여 0, 10, 30초, 1, 1.5, 2, 3, 4, 5, 7 및 10분 간격으로 대퇴동맥에 설치한 카테타로부터 혈액을 채취하고 마지막 혈액을 취한 후 즉시 단두하여 뇌를 적출하였다. 채취한 뇌를 액체질소에 담근 후 tissue Tek으로 고정하고 autoradiography를 하기 위한 slice와 농도측정을 위해 필요한 부위를 취한 후 감마카운터로 Tc-DTPA의 brain각 부위별 및 혈장농도를 측정하였다. 뇌조직중 혈액부피의 측정은 Tc-albumin를 이용하여 구하였다. 같은 방법으로 5-FU에 대한 실험도 행하였고, 이때 각 혈장 및 조직중의 5-FU함량은 HPLC로 하였다.

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Partial purification and characterization of phosphatidylcholine hydrolyzing enzyme from liver membrane of flounder , Paralichtys olivervaceus (넙치 간에 있어 가수분해 효소의 부분정제 및 특성규명)

  • Lee, Sang-Hwan;Seo, Jeong-Su;Kim, Na-Yeong;Eom, Hye-Gyeong;Wi, Hyo-Jin;Park, Seong-Il;Jeong, Jun-Gi
    • Journal of fish pathology
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    • v.17 no.2
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    • pp.131-137
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    • 2004
  • In the present study, phosphatidylcholine (PC) hydrolyzing enzyme had been isolated from membrane of flounder liver. PC hydrolyzing enzyme solubilized in 1% Triton X -100 from membrane was partially purified by sequential chromatography on Heparin Sepharose CL-6B and Heparin-5PW columns. The products by membrane-bound hydrolyzing enzyme were identified as phosphatidic acid and choline, but in the presence of primary alcohol, phosphatidylethanol was produced at the expense of phosphatidic acid. These data suggest that membrane-bound enzyme may be a PC-phosphoipase D (PLD) type. The enzyme had pH optimum at below 6.0 and temperature optimum at $37^\circ{C}$. The activity of PC-PLD was dose-dependently increased by $Ca^{2+}$ but not $Mg^{2+}$. The activity of PC-PLD was stimulate by PC, PIP2 and PE.

Studies on the digestive gland structures of domestic and experimental animals II. The renewal cycle of the cells in the intestinal mocosa of sucking mouse (가축(家畜) 및 실험동물(實驗動物) 소화관선(消化管腺)의 구조(構造)에 관한 연구 II. 포유(哺乳) mouse 장점막(腸粘膜)의 세포재생주기(細胞再生週期)에 대하여)

  • Kwak, Soo-dong
    • Korean Journal of Veterinary Research
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    • v.32 no.1
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    • pp.1-6
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    • 1992
  • The turnover time of the mucosal epithelium in the small intestine(jejunum and ilium) and large interstine(cecum), and the cells in the lamina propria of the small intestine was investigated with the radioautography in mice at various times after single injection of $^3H$-thymidine. Twenty suckling mice were sacrified at each of the following time intervals after injection ; 2 hrs, 1, 3, 5. 7, 14 and 17 days. 1. The labeled index of the epithelial cells in the crypt and the villus of the small intestine averaged 98.7% and 1.3% at 2 hrs, 982% and 1.8% at 1 day, 18.7% and 81.3% at 3 days, 6.3% and 93.7% at 5 days, respectively. The labeled index of the epithelial cells of the crypt-base, the upper-crypt and the mucosal surface in the large intestine averaged 71.8%, 28.2% and 0% at 2 hrs, 45%. 54.2% and 0% at 1 day, 17.2%, 54.5% and 28.2% at 3 days, 10.2%, 32.4% and 57.4% at 5 days, respectively. This result suggested that the turnover time of all the epithelial cells migrating from crypts to villi in the direction of the villus tips was calculated to be less than 5 days, and also the longest turnover time was calculated to be no longer than 7 day. 2. The labeled index of the total cells in the lamina propria of the small intestine averaged 6.2-7% at 2 hrs to 5 days, 4.7% at 7 days 2.6% at 17 days and this index is tend to be decreased moderately at 7 days and severely at 17 days. So this result suggested that the turnover time of the cells with the shorter cycle duration in the lamina propria of the small intestine were less than 5 days and that of the cells with the longer cycle duration more than 17 days.

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Ethanol Tolerance of Campylobacter jejuni by Ethanol Shock (Ethanol 충격에 의한 Campylobacter jejuni 의 Ethanol 내성)

  • 김치경;가익현
    • Korean Journal of Microbiology
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    • v.30 no.5
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    • pp.377-382
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    • 1992
  • The responses of C. jejuni to ethanol shock were studied for their survival. synthesis of ethanol shock proteins, and increased survival at higher concentration of ethanol upon prior treatments of ethanol. When C. jejuni were shocked with ethanol at 1. 3. and 5% for 60. 30 and 10 minutes, respectively. those cells synthesized the ethanol shock proteins of 90, 66, 60, 45, and 24 kd in molecular weight. When the C. ,jejuni shocked with 1 and 3% ethanol were exposed to 3 and 5% ethanol for 30 minutes. their survival rates were increased by $10^1$~$10^2$ as compared with those of the cells without ethanol-shock. In the same way. C. ,jejuni shocked with 5% ethanol for 10 minutes :.bowed about 102 times higher survival rates than the cells without ethanol-shock. This result suggests that C jejuni shocked with I-5% ethanol for 10-30 minutes synthesized five kinds of ethanol shock proteins. and that the shock proteins contributed to increase ethanol tolerance for their survival at the higher concentrations of ethanol.

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ARACHIDONIC ACID METABOLISM IN HYPERSENSITIVE HUMAN DENTAL PULP (지각과민성(知覺過敏性) 치아(齒牙) 치수조직(齒髓組織)의 Arachidonic Acid Metabolism에 관(關)한 연구(硏究))

  • Lee, Kyung-Hee;Son, Ho-Hyun
    • Restorative Dentistry and Endodontics
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    • v.15 no.1
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    • pp.153-164
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    • 1990
  • Human dental pulps obtained from normal teeth, hypersensitive teeth and teeth with inflamed pulp were studied to measure and to compare the endogenous levels of arachidonic acid metabolites in order to see the relative activities of the different pathways involved in arachidonic acid metabolism. Pulp homogenates were incubated with $^{14}C$-arachidonic acid and lipid solvent extracts were separated by thin layer chromatography (TLC) to be analyzed by autoradiography and TLC analyzer. 1. The most significant metabolite was HETEs showing $96.9{\pm}37.8$pmol/mg tissue protein/hr in normal pulp, $169.2{\pm}76.7$ in hypersensitive pulp and $385.4{\pm}113.2$ in inflamed pulp. In normal pulp $LTB_4$, 6-keto-$PGF_{1\alpha}+PGE_2$, $TXB_2$ and unidentified metabolite were formed in decreasing order. While in hypersensitive and inflamed pulp 6-keto-$PGF_{1\alpha}+PGE_2$, $LTB_4$, $TXB_2$ and unidentified metabolite were formed in decreasing order. 2. In hypersensitive pulp only HETEs were significantly increased when compared with that in normal pulp. The levels of all the converted metabolites in inflamed pulp were significantly increased compared with those in normal pulp. In inflamed pulp, the levels of $TXB_2$ and HETEs were significantly increased compared with those in hypersensitive pulp. 3. The ratio of each metabolites to the total converted metabolites showed an increased value of $TXB_2$ and 6-keto-$PGF_{1\alpha}+PGE_2$, as the degree of inflammation was increased, while that of HETEs decreased both in hypersensitive pulp and inflamed pulp more than in normal pulp. 4. The relative amounts of the total metabolites formed in lipoxygenase pathway to cyclo-oxygenase pathway were 6.8 fold in normal pulp, 4.4 fold in hypersensitive pulp and 3.8 fold in inflamed pulp.

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Attenuation of Morphine Tolerance and Withdrawal Syndrome by Coadministration of Nalbuphine

  • Jang, So-Yong;Kim, Hee-Jeong;Kim, Dong-Hyun;Jeong, Myeon-Woo;Ma, Tangen;Kim, Seong-Youl;Ho, Ing K.;Oh, Sei-Kwan
    • Archives of Pharmacal Research
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    • v.29 no.8
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    • pp.677-684
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    • 2006
  • Morphine has been used widely on the treatment of many types of chronic pain. However the development of tolerance to and dependence on morphine by repeat application is a major problem in pain therapy. The purpose of the present study was to investigate whether combined administration of nalbuphine with morphine affects the development of tolerance to and dependence on morphine. We hypothesize that the use of nalbuphine, ${\kappa}-agonist$ may prove to be useful adjunct therapy to prevent morphine-induced undesirable effects in the management of some forms of chronic pain. Morphine (10 mg/kg) was injected to rats intraperitoneally for 5 day. The variable dose of nalbuphine (0.1, 1.0 and 5.0 mg/kg) was administered (i.p.) in combination with morphine injection. The development of morphine tolerance was assessed by measuring the antinociceptive effect with the Randall-Selitto apparatus. The development of dependence on morphine was determined by the scoring the precipitated withdrawal signs for 30 min after injection of naloxone (10 mg/kg, i.p.). Nalbuphine did not attenuate antinociceptive effect of morphine in rats. Interestingly, combined administration of morphine with nalbuphine (10:1) significantly attenuated the development of dependence on morphine. The elevation of $[^3H]MK-801$ binding in frontal cortex, dentate gyrus, and cerebellum after chronic morphine infusion was suppressed by the coadministration of nalbuphine. In addition, the elevation of NR1 expression by morphine was decreased by the coadministration of nalbuphine in rat cortex. These results suggest that the coadministration of nalbuphine with morphine in chronic pain treatment can be one of therapies to reduce the development of tolerance to and dependence on morphine.

Effects of Anti-Neoplastic Antibiotics on DNA Replication and Repair (DNA복제 및 회복에 미치는 수종항암 항생제의 영향에 관한 연구)

  • Park, Sang-Dai;Rie, Myung-Chull;Lee, Chun-Bok
    • The Korean Journal of Zoology
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    • v.26 no.1
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    • pp.19-28
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    • 1983
  • Alkaline elution profiles showed that the frequency of DNA single strand breaks associated with DNA-protein crosslinks in cells treated with both an inducing dose of MMC $(MMC_1)$ and a challenge dose of MMC $(MMC_2)$ was slightly less than that in cells treated with MMC alone. The amount of unscheduled DNA synthesisi in cells treated with both $MMC_1$ and $MMC_2$ was greater than that in cells treated with MMC alone. This enhancement of exicision repair detected by UDS autoradiography and alkaline elution, was not observed, when cells were incubated with cyclohexmide between the two treatments of $MMC_1$ and $MMC_2$. These results suggest that MMC-damaged DNA from Chinses hamster cells is repaired by excision repair mechanisms that require de novo protein synthesis for enhancement, and that an inducible repair mechanism may exist in CHO cells.

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A STUDY ON ARACHIDONIC ACID METABOLISM OF CHRONIC PERIAPICAL LESIONS (만성 치근단주위 병소조직의 Arachidonic acid 대사에 관한 연구)

  • Park, Keum-Soon;Son, Ho-Hyun
    • Restorative Dentistry and Endodontics
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    • v.17 no.1
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    • pp.83-94
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    • 1992
  • This study was executed to measure the biosynthesis of arachidonic acid metabolic products in chronic periapical lesions, to compare the products among periapical granuloma, periapical cyst and chronic periapical abscess, and to understand the pathogensis of chronic periapical lesions. Tissues from 33 chronic periapical lesions of human teeth were enucleated during endodontic surgery. large part of each tissue was contained in liquid nitrogen immediately and the other was examined histologically. In histologically diagnosed 8 cases of periapical granuloma, 9 cases of periapical cyst and 8 cases of chronic periapical abscess. the tissues were homogenatecl and incubated with $_{14}C$-arachidonic acid. Lipid solvent extracts were separated by thin layer chromatography to be analyzed by autoradiography and TLC analyzer. 1. $TXB_2$, 6-keto-$PGF_1{\alpha}$ and $PGE_2$, $LTB_4$, HETEs, and unidentified product which are metabolic products of arachidonic acid were measured in the tissues of chronic peripaical lesions. 2. In all of periapical granuloma, cyst and abscess, the conversion rate of HETEs among all products was the highest(P<0.05), and the percentage of HETEs in total converted products was also the highest(P<0.05). 3. The concentration of each arachidonic acid product was higher in chronic periapical absecss than in periapical granuloma and cyst(P<0.05). The concentration of $TXB_2$ and HETEs in periapical cyst were hight than in periapical granuloma. 4. The relative amounts of total products from lipoxygenase pathway to those from cyclo-oxygenase pathway were about 7 fold in chronic periapical lesions. There was no difference among periapical granuloma, cyst and abscess(P<0.05). The total amount of products from each pathway were higher in chronic periapical abscess than in periapical cyst and granuloma.

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Characterization of Thyrotrpin Releasing Hormone Receptor in Brain (뇌의 Thyrotropin Releasing Hormone 수용체 분포에 관한 연구)

  • Koong, Sung-Soo;Kim, Seung-Taik;Chung, June-Key;Lee, Myung-Chul;Cho, Bo-Youn;Koh, Chang-Soon;Yoo, Eun-Sook;Park, Eun-Mey;Seo, Il-Tak
    • The Korean Journal of Nuclear Medicine
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    • v.24 no.1
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    • pp.93-100
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    • 1990
  • To evaluate the utility of autoradiographic technique in the detection of TRH receptor changes in brain after the various kinds of stimulation or drug administration, we tried the characterization of TRH receptor in mouse brain and autoradiography in rat brain as a preliminary study. The Kd value of [3-H] MeTRH to TRH receptors of adult male ICR mouse brain (cebellum and spinal cord were excluded) was 3.55+0.6 nM and Bmax was 3.44+0.52 fmol/mg wet tissue by saturation analysis. The Kd value of TRH to TRH receptors was 133.8+28.2 nM by competition analysis. And we could visualize the distribution of TRH receptors in rat brain by autoradiographic technique.

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Effect of cooling rate on the hot ductility of boron bearing steel during continuous casting (Study for prevention of corner crack on continuous casting slab) (보론 첨가강에서 연주 냉각속도가 고온연성에 미치는 영향 연구 (주편 코너 크랙 발생 방지 방안 확보 연구))

  • Cho, Kyungchul;Koo, Yangmo;Park, Joongkil
    • Korean Journal of Metals and Materials
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    • v.46 no.6
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    • pp.329-337
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    • 2008
  • During the continuous casting of boron-bearing steel, the corner cracks on the slab are formed by deformation with low strain rate and rapid cooling at the unbending temperature within the range of 800- $1000^{\circ}C$. Especially, the rapid cooling in the corner of slab during the continuous casting leads to as corner cracking. Therefore, in this study, the hot tensile tests applied to the different cooling rates were taken into account in order to study the effect of cooling rate on the hot ductility of boron-bearing steel. The results revealed that increasing cooling rate deteriorate the hot ductility of boron- bearing steel. Rapid decreasing of the hot ductility is caused by formation of a film-like ferrite and precipitate at the austenite grain boundaries. The morphology of the precipitates in the boron-bearing steel was monitored by PTA (Particle Tracking Autoradiography) and TEM, we observed MnS and BN compound and their morphology was quite different depending on the cooling rates. When the cooling rate is increased, rodshape MnS and BN precipitates can be formed along the austenite grain boundaries. It can cause that weakening the boundary region and decreasing the hot ductility of boron-bearing steel.