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Genotoxicity on Structural Derivatives of Sophoricoside, a Component of Sophora Japonica, in Bacterial and Mammalian Cells

  • Ryu, Jae-Chun;Kim, Youn-Jung;Kim, Mi-Soon;Kim, Min-Ji;Sarma, Sailendra Nath;Jung, Sang-Hun
    • Molecular & Cellular Toxicology
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    • v.1 no.3
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    • pp.179-188
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    • 2005
  • To develop the novel anti-allergic drug, many sophoricoside derivatives were synthesized. Among these derivatives, JSH-II-3, VI-3, VII-3, VIII-3, VII-20 and VII-20 (sodium salt) were selected and subjected to high throughput toxicity screening (HTTS) because they revealed strong IL-5 inhibitory activity and limitation of quantity. Single cell gel electrophoresis (Comet) assay, mouse lymphoma thymidine kinase ($tk^{+/-}$) gene assay (MOLY), chromosomal aberration assay in mammalian cells and Ames reverse mutation assay in bacterial system were used as simplified, inexpensive, short-term in vitro screening tests in our laboratory. Through the primary screening using the comet assay, we could choose the first candidates of sophoricoside derivatives with no genotoxic potentials as JSH-VI-3, VII-3, VII-20 and VII-20 (sodium salt). Also JSH-VII-3, VII-20 and VII-20 (sodium salt) are non-mutagenic in MOLY assay, while JSH-II-3 is mutagenic at high concentration with the presence of metabolic activation system in both comet assay and MOLY assay. The selected derivatives (JSH-VI-3, VII-3, VII-20 and VII-20 (sodium salt) are not mutagenic in S. typhimurium TA98 and TA100 strains both in the presence and absence of metabolic activation. From results of chromosomal aberration assay, 6 h treatment of JSH-VI-3, VII-3 and VII-20 (sodium salt) were not revealed clastogenicity both in the presence and absence of S-9 mixture. Therefore, we suggests that JSH-VI-3, VII-3, VII-20 and VII-20 (sodium salt), as the optimal candidates with both no genotoxic potential and IL-5 inhibitory effects must be chosen. To process the development into new anti-inflammatory drug of these derivatives, further investigation will need.

In Vivo Tumor Cell Distribution of Antibody-Endostatin Fusion Protein for Tumor-Specific Targeting and Pharmacokinetics (암세포 표적지향화를 위한 항체-엔도스타틴 융합단백질의 체내동태 및 종양으로의 이행성)

  • Kang, Young-Sook;Lee, Na-Young
    • Journal of Pharmaceutical Investigation
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    • v.33 no.4
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    • pp.287-292
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    • 2003
  • A novel antitumor agent, antibody-endostatin fusion protein $(anti-HER2/neu\;IgG3C_H3-Endostatin,\;AEFP)$ formed by genetic engineering procedure from antibody (Ab) which specifically targets to tumor cells ad angiogenesis inhibitor, endostatin (Endo) that has excellent antitumor effect, minimizes the toxicity of normal cells and selectively kills only tumor cells. The purpose of this study is to evaluate the phamacokinetic parameters and to analyze the localization of AEFP. After an intravenous injection of $150\;{\mu}l\;(5\;{\mu}Ci)\;[^{125}I]Ab,\;[^{125}I]AEFP$ to mice, blood was collected though retroorbital plexus from 15 min to 2880 min. Following the jugular vein injetion of $150\;{\mu}l\;(10\;{\mu}Ci)\;[^{125}I]Endo$, blood was collected by the use of carotid artery cannulation from 0.25 min to 30 min. Consequently, Endo was very rapidly removed from plasma compartment within 30 min. On the other hand, AEFP similar to Ab was slowly cleared from plasma. Also, Endo was metabolized about 40% within 30 min. However, AEFP was shown to metabolize less than 10% within 2880 min. The organ distribution of Endo was in order kidney, lung, spleen. Both Ab and AEFP were localized in order spleen, kidney, liver. Futhermore the tumor/blood distribution ratio of AEFP at 96 hours after injection is about 20 times higher than it of Endo at one hour after injection. In conclusion, these studies demonstrate that the anti-cancer or suppression of angiogenesis effect of Endo may be improved by the use of AEFP because the longer half life and stability of AEFP is able to selectively target antigens expressed on tumors.

Chondroblastoma of the Base of Fifth Metatarsal Bone associated with Secondary Aneurysmal Bone Cystic Change - A Case Report - (제 5 중족골 기저부의 이차 동맥류성 골낭종변화를 동반한 연골모세포종 - 증례 보고 -)

  • Son, Hyun-Jin;Jang, Kyu-Yun;Lee, Dong-Geun;Lee, Sang-Yong;Kim, Jung-Ryul
    • The Journal of the Korean bone and joint tumor society
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    • v.10 no.1
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    • pp.45-49
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    • 2004
  • Chondroblastoma is a benign chondroid-forming tumor usually originating in the epiphysis. The base of metatarsal bone contains neither an epiphysis nor a secondary ossification center and so is the rare site of chondroblastoma. Here, we present a case of chondroblastoma of the base of fifth metatarsal bone in 34-year-old man. Histologically, the osteoclast-like giant cells were abundant enough to simulate a giant cell tumor. And the chondroid intercellular matrix was intermixed but scanty. However, the background mononuclear cells showed irregular and indented nuclei with longitudinal clefts and positive immunoreactivity for S-100 protein, as the evidence of chondroblasts.

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Sex Differences in the Association of Habitual Snoring with Metabolic Syndrome (성별에 따른 습관성 코골이와 대사증후군 요소의 관련성)

  • Kim, Seong Joon
    • The Journal of the Korea Contents Association
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    • v.14 no.1
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    • pp.326-336
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    • 2014
  • The purpose of this study was to assess the association between habitual snoring and metabolic syndrome in the Korean population and to evaluate it's possible gender variations. The study subjects were 2,007 volunteers aged 40-69 years who participated in regular health check-ups(male:434, female:1,573) from May 2012 to December 2012. Standard interviews, anthropometrics and biochemical studies were conducted. Habitual snoring was defined as snoring more than 4 days per week. The prevalence of metabolic syndrome in our sampling pool was 17.4% (male: 20.7%, female: 16.5%), and this increased with age. After adjusting for age, smoke and alcohol consumption, sleep duration, regular exercise, the odds ratio of metabolic syndrome and it's components were increased significantly by habitual snoring. HDL-cholesterol, blood sugar, waist size were significantly higher in women, but not significantly higher in men.

Change in Storage Protein during Germination of Peanut Seed (땅콩발아시 저장단백질의 변화)

  • 김종진;박재욱;신동현
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.39 no.6
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    • pp.564-570
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    • 1994
  • This experiment was conducted to determine seed storage protein pattern and structural character of differed peanut cultivars during germination. Soluble protein content in both Namdae and Daekwang cultivars remarkably decreased in cotyledon site at 2 or 3 days after incubation(DAI) and in embryonic axis site at 1 or 2 DAI, showing 28∼29% in cotyledon site and 10% in embryonic axis site at 5 DAI. Protein subunits such as 66, 43, 40 and 35.5kD bands in the cotyledon site of Namdae and Daekwang cultivars disappeared, but 21.5-23kD band disappeared slightly, but low polypeptide band such as 14-16kD increased gradually, and the same trend has been obserbed in embryonic axis site during 2 DAI. The amount of new protein formed during germination period was highest in cotyledon site at 3 DAI, and in embryonic axis site at 2 DAI. 16kD bend detected in cotyledon site of Daekwang cultivar during germination.

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Growth of Yeasts in Alcohol Distiller′s Waste of Dried Sweet Potato for Single-cell Protein Production and BOD Reduction (절간고구마원료 주정폐액을 이용한 단세포단백질의 생산 및 폐액의 BOD제거)

  • 이형춘;구영조;민병용;이홍근
    • Microbiology and Biotechnology Letters
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    • v.10 no.2
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    • pp.95-100
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    • 1982
  • Torulopsis candida FRI YA-15, a selected yeast, was cultivated in alcohol distiller's waste-filtrate of dried sweet potato for microbial protein production and BOD reduction. The General composition of waste-filterate was BOD$_{5}$ 15700 ppm, COD 36800 ppm, reducing sugar 3300 ppm, total nitrogen 910 ppm, total solids 51800 ppm and ash 390 ppm. The pH of waste was 3.85. The yield to the medium of T. candida cultivated in shake-flask at $25^{\circ}C$ for 48 hrs was 3.38g/$\ell$ and effectiveness in reducing BOD$_{5}$ and COD of waste was 38.9% and 31.8%, respectively. In batch cultivation using 3 $\ell$-jar fermenter, maximum yield to the medium reached 3.2g/$\ell$after 28 hrs cultivation under the condition of temperature 35$^{\circ}C$, initial pH 4.0, aeration rate 2vvm, agitation speed 100rpm. Dry yeast was composed of crude protein 47.98% and ash 5.23%.

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Two-Dimensional Electrophoresis를 이용한 한우 난소의 황체단백질 특성 분석

  • 우제현;정학재;김봉기;최재혁;박민영;양병철;박수봉;성환후;권무식
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.85-85
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    • 2003
  • 한우 난소의 황체는 다양한 세포들로 구성되어 있으며 난소의 생식기능유지와 임신유지에 중요한 인자가 복잡하게 관련되어 있으며 이들 황체에서 분비하는 단백질은 황체기능에 필수적으로 중요한 작용을 한다. 본 연구는 난소의 황체일령에 따른 단백질 분비 패턴을 조사함으로써 황체세포의 기능과 임신 유지에 관련되는 인자들을 조사하기 위하여 수행하였다. 한우에서 채취한 난소에서 황체를 분리, 황체시기별(전기, 중기, 말기)로 구분하여 cytosol을 분리 정제하였다. 황체시기별로 분리된 황체 단백질의 성분을 분석하기 위해 ion-exchange chromatography를 이용하여 단백질 패턴을 조사, 추출된 fraction을 단백질 정량후 SDS-PAGE를 실시하였다. 그 결과 중기에서의 단백질의 농도가 가장 높았으며, 특히 단백질 패턴 또한 다른 양상을 보였다. 시기별로 구분한 각각의 fraction을 SDS-PAGE로 조사했을 때 중기와 말기황체 cytosol의 fraction 3번과 4번에서 다른 양상을 보였으며 SDS-PAGE에서 120kb, 95kb, 34kb, 25kb 등의 단백질 밴드를 확인할 수 있었다. 초, 중기황체에서만 특이하게 검출되는 단백질 band를 확인할 수 있었으며 이 들 단백질을 구체적으로 확인하기 위하여 Two-Dimensional electrophoresis를 이용하여 실험을 수행하였다. 시기별로 분리된 황체를 일반적인 IEF단백질 분리법으로 cytosol을 회수한 후 IPG-system을 이용하여 1차원 전기영동을 한 후, SDS-PAGE로 이차원 전기영동을 실시하였다. 이차원 전기영동 결과, SDS-PAGE의 결과와 비슷한 위치에 부분적으로 다른 spot의 양상을 보였다. 특히 기능황체에서의 특이적 발현 spot을 확인할 수 있었다. 이러한 결과들로부터 황체의 progesterone분비기능의 역할을 수행하기 위한 단백질들이 전, 중기에 발현된다는 것을 알 수 있고 퇴행황체에서는 발현이 안되고 있는 것을 알 수 있다. 이러한 결과를 토대로 다른 양상을 띤 spot을 분리하여 어떤 단백질인지를 분석하여 각각의 황체단백질의 특성을 규명할 수 있을 것으로 사료된다.

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Mobilization of Proteins in the Cotyledons of Germinating Soybeans(Glycine max) (발아중인 대두 (Glycine max)에서의 단백질 유동)

  • Song, Young-Sun
    • Korean Journal of Food Science and Technology
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    • v.20 no.5
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    • pp.650-658
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    • 1988
  • The mobilization of proteins in the cotyledons of germinating soybean seeds (Glycine mar [L.] Merr.) and seedlings was studied by using light microscopy and transmission electron microscopy. The cotyledon tissues of soybean. were packed with protein bodies(diameter $0.1-15{\mu}m$) where storage protein of soybean is deposited. Degradation of protein bodies started in the epidermis and vascular tissues. After swelling of the protein bodies, autolysis of storage proteins began while the external membrane remained unbroken. Hydrolysis of proteins could be internal or peripheral and fusion might begin before complete protein degradation. Possible instances of vacuolar fusion were encountered in some cells. In all cases, the result of degradation was the same; the central vacuole of the cell. At the late stages of seedling growth, breakdown of tonoplast was observed in some cells.

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Role of Samultang in Formalin-induced Orofacial pain (포르말린으로 유발된 안면부 통증조절에서 사물탕의 역할)

  • Kim, Yun-Kyung;Hyun, Kyung-Yae;Park, Min-Kyoung;Lee, Min-Kyung
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.15 no.9
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    • pp.5594-5601
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    • 2014
  • The aim of this study was to evaluate the analgesic effects of Samultang by measuring the anti-inflammatory and antioxidative activities in formalin-induced orofacial pain. The anti-inflammatory and anti-oxidative effects were measured by western blot analysis for p38 MAPK(Mitogen-activated protein kinases), iNOS(induceble nitric oxide synthase) and NOX4(nicotinamide adenine dinucleotide phosphate4) in a rat's brain and medulla oblongata. Samultang significantly attenuated the increased formalin-induced nociceptive response and inhibited the p38 MAPK and iNOS that led to inflammatory pain. In addition, this study indicated that Samultang possess antioxidative effects through the blockade of NOX4. These findings suggest that Samultang plays an important role in the peripheral processing of inflammatory pain in the orofacial area.

Analysis of outer mombrane proteins of Brucella abortus using two dimensional polyacrylamide gel electrophoresis (2차원 전기영동법을 이용한 Brucella abortus 세포외막 특이단백질의 분석)

  • Kim, Byung-su;Kim, Sun-hee;Kim, Jong-suk;Baek, Byeong-kirl
    • Korean Journal of Veterinary Research
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    • v.38 no.2
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    • pp.328-335
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    • 1998
  • Outer membrane proteins(OMPs) of Brucella abortus 1119-3 strain were extracted by Triton X-100 treatment, and fractionated by DEAE-cellulose column chromatography and Sephacryl S-300 column chromatography. The antigenic proteins in these fractions were identified by Western blot analysis. In Western blot analysis, a single band(38kDa) was observed in the DEAE fractions from 36th fraction to 38th fraction against sera of cattle infected with B abortus. And other fractions have several bands. However, the Sephacryl S-300 fractions exhibited a total of 3 peaks of proteins with a broad range from about 30 to 116kDa. In order to characterize further, the extracted OMPs and the DEAE fractions were analyzed by two dimensional polyacrylamide gel electrophoresis(2-DE) and Western blot using serum from naturally infected cattle with Brucella spp. The 2-DE immunoblots of DEAE fraction showed immunoreactive spots more than twenty two. The major protein spots have ranging from about 32 to 47kDa. The pI values of the spots were detected from pH 4.7 to 5.4. Among the major protein spots, the 38kDa protein which is a specific antigen, located at the point of approximately a pI 4.8.

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