• Title/Summary/Keyword: protein gel

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Protein Composition and Biological Activities of Bombus ignitus Venom (호박벌 일벌독의 성분 분석 및 생리활성 탐색)

  • Han, Sang-Mi;Lee, Kwang-Gill;Yeo, Joo-Hong;Kweon, Hae-Yong;Woo, Soon-Ok;Yoon, Hyung-Joo;Kim, Me-Ae;Kim, Won-Tae;Baek, Ha-Ju
    • Korean journal of applied entomology
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    • v.46 no.2
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    • pp.229-234
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    • 2007
  • Pure Bombus ignitus venom samples were submitted to two-dimensional gel electrophoresis. A total of 64 excised spots were analyzed by mass spectrometry. Three main proteins resulted in the identification have not been described in other bee venoms before. Dose-dependence against human carcinoma (Hep3B, BT-20, A549 and AGS) were observed from 1ng/ml to 100ng/ml. Expecially, the treatment of 100ng/ml B. ignitus venoms showed the highest cytotoxicity with 55% against hepatocellular carcinoma (Hep3B). The B. ignitus venoms showed strong antimicrobial activities against Enterococcus faecium and Shigella sonnei, and practically antimicrobial activity against the other microorganisms tested. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of E. faecium and S. sonnei, were 0.256ug/ml, respectively.

Pharmacokinetics of Recombinant Human Epidermal Growth Factor (DWP401) after Repeated Subcutaneous Administration to Rats (흰쥐에 재조합 인간 상피세포 성장인자(DWP401)를 연용피하투여했을 때 약물체내동태)

  • Nam, Gwon-Ho;Jo, Jae-Yeol;Jeong, Ju-Yeong;Jang, U-Ik;Gang, Jin-Seok;Yu, Eun-Suk;Park, Seung-Guk;Yu, Yeong-Hyo;Park, Myeong-Hwan;Sim, Chang-Gu
    • YAKHAK HOEJI
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    • v.40 no.5
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    • pp.491-500
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    • 1996
  • The organ distribution and pharmacokinetics of DWP401, a recombinant human epidermal growth factor (rhEGF), were compared after single and repeated subcutaneous administration ( 50${\mu}$/kg, 10${\mu}g$Ci/kg of $^{125}I$-DWP401, twice a day for 7 consecutive days) to rats. The pharmacokinetic parameters such as AUC and terminal half-life were similar between two different administration. During repeated administration, the plasma concentration of DWP401 seemed to be constant when the plasma was collected at 15 min after each dosing. The TCA-precipitated radioactivities in thyroid, liver, kidney, and stomach were higher than those of other organs studied after both single and repeated administration. The TCA-precipitated radioactivities after repeated administration in several organs, such as thyroid, stomach, prostate, adrenal, eye ball, and testis were higher than those after single administration. But, according to the observations using gel filtration chromatography and antibody binding assay, the radioactivities in thyroid and stomach were not primarily due to the intact DWP401 or its metabolites but due to the $^{125}I$-thyroxine binding protein. In conclusion, it can be suggested that DWP401 is metabolized to each amino acid or small polypeptides, and there was no significant changes in pharmacokinetics or any indications for accumulation of DWP401 in rat plasma and organs after repeated treatment.

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Comparison of Food Quality between Finespotted flounder and Their Similar Kinds for Material Distinction in Raw Fish Sliced with Bones(small sashime or sekoshi) (뼈째썰기회의 원료판별을 위한 도다리와 유사어종과의 식품학적 특성비교)

  • Kim, Sung-Hun;Kang, Hyun-Woo
    • Culinary science and hospitality research
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    • v.19 no.5
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    • pp.158-169
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    • 2013
  • Finespotted flounder, used for a representative raw fish in spring, is considered comparatively difficult to breed, which causes small Olive flounder and Stone flounder from China, similar kinds of flounders, to appear on the market for sale under the name of 'Finespotted flounder.' The reason lies under the considerations that small Olive flounder and Stone flounder from China are relatively lower priced and in higher supply and demand, being difficult to distinguish from Finespotted flounder when sliced with bones. Thus, the purpose of our thesis is to distinguish Finespotted flounder from similar kinds of fish analyzing the lipid content in slices of raw fish and SDS-PAGE(sodiumdodecylsullipide-polyacrylamide gel eletrophoresis). Upon comparing the main components between sliced Finespotted flounder and similar kinds of fish, such as small Olive flounder and Stone flounder from China, we found that there are no noticeable differences among them in moisture content, and little, if any noticeable differences, in crude protein and ash content(P>0.05). Based on these analyses, we have conclude that commercial raw fish restaurants sell small Olive flounder and Stone flounder under the name of Finespotted flounder. However, a variety of factors have an effect on our analysis, such as the individual characteristics of fish and a seasonal variation. The aim of our analysis is to enhance more accurate distinction criteria, although some fish kinds can be discerned with our present technique of examining lipid content and SDS-PAGE. Through more sophisticated analyses developed by consistent research, we look forward to attaining more accurate techniques for discerning between Finespotted flounder and different kinds of similar fish.

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A Study on the Antitumor Activity of Panax ginseng (고려인삼의 항암효과에 관한 연구)

  • Hwang, Woo-lk
    • Journal of Ginseng Research
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    • v.17 no.1
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    • pp.52-60
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    • 1993
  • Panax ginseng has been extensively used in the traditional oriental medicine as a restorative, tonic and Prophylactic agent. Recently, several reports regarding to anticancer effects of Panax ginseng has accumulated. These studies emphasized the fact that the anticancer activities might be due to a glycoside group called ginsenoside or pan.u saponin which has a water soluble characteristic. However, the authors and collaborates demonstrated that a highly lipid soluble component in extract of Panax ginseng roots contains a considerable cytotoxic activities against marine leukemic cells (L1210, P388) and human censer cells (HRT-18, HT-29, HCT48). This study was devised to observe the cytotoxic activities of Petroleum-ether extract of Panax giuseng roots (crude GBD and its Partially Purified fraction from silicic acid column chromatography (7 : 3 GX) against sarcoma-180 (5-180) and Walker carcinosar- coma 256 (Walker 256) in vivo, and murine leukemic Lymphocytes (L1210) and human rectal cancer cells (HRT-18) and human colon cancer cells (HT-29 and HCT48) in vitro. Each cell-line was cultured in medium containing serial concentration of the crude GX or 7 : 3 GX in vitro. A highly lipid soluble compound in the extract of Panax ginseng root was cytocidal to murine leukemic cells and human colon and rectal cancer cells in vitro. In the meantime, ginseng saponin derivatives did not have cytotoxic effects at its corresponding concentration. The growth rates of the cancer cells in medium containing ginseng extracts were inhibited gradually to a significant degree roughly in proportion to the increase of the extract concentration. The cytotoxic activity of 7 : 3 GX was about 3 times more potent than that of crude GX, one unit of cytotoxic activity against L1210 cells being equivalent to 2.54 Ug and 058 Ug for the crude GX and 7 : 3 GX, respectively. The Ri value of the active compound on silica- gel thin layer chromatography with petroleum-ether/ethyl ether/acetic acid mixture (90 : 10 : 1, v/v/v) as a developing so lvent was 053. While, the Panaxydol and Panaxynol as active compounds were purified from Petroleum-ether extract of Panax ginseng root by Drs. Ahn and Kim, and author found out that the one unit of cytotoxic activity of the Panaxydol and Panaxynol against L1210 cells being equivalent to 056 Ug and 0.3918 respectively. The survival times of mice inoculated with S-180 cells were extended about 1.5 to 2 times by the 7 : 3 GX treatment compared with their control group. The significantly decreased hemoglobin values of rats after inoculation with Walker 256 were recovered to normal range by oral administration of the crude Gt The synthetic levels of protein, DNA and RNA in human colon and rectal cancer cells were significantly diminished by treatment with the crude GX, which can explain a part of the origin of its anticancer activity.

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Purification and Characterization of ($Ca^{2+}$+$Mg^{2+}$)-ATPase of Sarcoplasmic Reticulum from Rat Skeletal Muscle (쥐 근소포체의 ($Ca^{2+}$+$Mg^{2+}$)-ATPase의 분리정제와 그 효소특성에 관하여)

  • Lee, Jong-Soon;Ha, Doo-Bong;Chung, Chin-Ha
    • The Korean Journal of Zoology
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    • v.28 no.1
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    • pp.31-43
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    • 1985
  • The $(Ca^{2+}+Mg^{2+})$-ATPase has been purified homogeneously from sarcoplasmic reticulum of rat skeletal muscle by sucrose density gradient centrifugation. The purified enzyme has a molecular weight of 115,000 as judged by polyacrylamide gel electrophoresis in the presence of sodium dedecyl sulfate, and therefore has the same size of the enzyme in rabbit and chick skeletal muscle. $Ca^{2+}, Mg^{2+}, Fe^{2+}, Co^{2+}, and Mn^{2+}$ at 50 $\\muM$ show stimulatory effect on the ATP-ase, while $Zn^{2+}, Cu^{2+}, and Hg^{2+}$ inhibit it at the same concentration. The ATPase activity is insensitive to antimalarial drugs such as quinine and quinacrine, but is sensitive to inhibition by p-hydroxymecurie benzoate and phenylmethylsulfonylfluoride. The enzyme has optimum pH of 6 to 7 and Km value for ATP is estimated to be 98 $\\muM$. Thus, a number of biochemical properties of this enzyme appear to be different from those of the enzyme that have been isolated from rabbit skeletal muscle. The $(Ca^{2+}+Mg^{2+})$-ATPase appears to be selectively degraded in microsomal fraction. The activity of metalloendoprotease is evident in the microsomal preparation when assayed by radioactively labeled protein substrate, such as $^{3}H-casein and $^{125}I$-insulin. However, it is presently unclear whether the metalloendoprotease is responsible for the degradation of the $(Ca^{2+}+Mg^{2+})$-ATPase.

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Fibrinolytic Activities and Effects of Gamma-Irradiated on Seeds from Coix lacryma-jobi L. Carthamus tinctorius L. and Malva verticillata L. (율무, 홍화, 아욱종자의 혈전용해 효소활성 및 감마선 조사의 영향)

  • Kwon Su-Jung;Lim Chae-Young;Kim Jae-Sung;Park Min-Hee;Lee Sook-Young
    • KSBB Journal
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    • v.21 no.1 s.96
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    • pp.20-27
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    • 2006
  • The fibrinolytic activities of soluble proteins extracted from seeds of Coix lacryma-jobi L., Carthamus tinctorius L. and Malva venicillata L. were studied. Fibrinolytic activity of extract from C. lacryma-jobi L. showed 1.3 times higher than plasmin used as positive control. The fibrinolytic enzyme was confirmed and extracted directly from seed of C. lacryma-jobi L. by a fibrin zymography. The protein was composed of a single polypeptide and its apparent molecular weight was found to be 7.8 kDa, as judged by SDS-polyacrylamide gel electrophoresis. The effect of temperature for the proteolytic enzyme activity were stabilized above $50^{\circ}C$ and then dramatically decreased. Also, the enzyme activity was clearly inhibited by APMSF, PMSF and TPCK, suggesting that it is a member of the chymotrypsin-like serine pretense. In addition, effects of gamma-irradiated on seed of each plants were revealed that 8 Gy and 64 Gy were higher than others. This result shown that gamma-irradiation of seeds were capable to increase the fibrinolytic activity. All these results suggest the pretense is a fibrinolytic enzyme belong to a family of chymotrypsin-like serine pretense.

Functional and Immunological Properties of Ryanodine Receptor in the Eel Skeletal Muscle (뱀장어 근육내 Ryanodine Receptor의 기능 및 면역학적 성질)

  • Seok, Jeong-Ho;Lee, Yeon-Soo;Nam, Jang-Hyeon;Choi, Suk-Jeong;Hong, Jang-Hee;Lee, Jae-Heun
    • The Korean Journal of Pharmacology
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    • v.31 no.2
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    • pp.207-217
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    • 1995
  • To investigate the functional and immunological properties of the Ca-release channel in the sarcoplasmic reticulum(SR) of the eel skeletal muscle, $[^3H]ryanodine$ binding, SDS gel electrophoresis, $^{45}Ca\;release$ studies, and immunoblot assay were carried out in the SR of the eel skeletal muscle. Maximal binding sites(Bmax) and $K_D$ values of $[^3H]ryanodine$ for Ca-release channel of the SR of the eel skeletal muscle were $19.44{\pm}1.40\;pmole/mg$ protein and $15.55{\pm}1.69\;nM$, respectively. $[^3H]Ryanodine$ binding to RyR was increased by calcium and AMP. The SR of the eel skeletal muscle has two high molecular weight bands on the SDS PAGE. The mobility of upper band was more slower than the single band of the rabbit skeletal muscle, and that of the lower band was similar with the single band of canine cardiac muscle. Vesicular $^{45}Ca-release$ was activated by calcium. Ca-induced $^{45}Ca-release$ was significantly inhibited by $MgCl_2(2\;mM)$, ruthenium red$(10\;{/mu}M)$ or tetracaine(1 mM), but not by high concentration of calcium itself. AMP-induced $^{45}Ca-release$ was slightly occurred only in the absence of calcium, it was not inhibited by $MgCl_2$ or ruthenium red. Caffeine also increased $^{45}Ca-release$ from the SR vesicles, but it was not affected by $MgCl_2$ or ruthenium red. Polyclonal Ab against rat skeletal muscle RyR is reacted with that of rabbit, but not reacted with that of the eel skeletal muscle. These results suggested that ryanodine receptor of the SR of the eel skeletal muscle is showing some similar properties with that of mammalian skeletal muscle, but might be an another isotype channel having two bands which is less sensitive to AMP, not cross-reacted with antisera against rat RyR, and not inhibited by high concentration of calcium.

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Some Properties of an Isolate of Peanut stunt virus Isolated from White Clover (Trifolium repens L.) (토끼풀에서 분리한 Peanut stunt virus의 성질)

  • Jung, Goo-Ho;Jeon, Yong-Woon;Choi, Jang-Kyung;Hong, Jin-Sung;Ryu, Ki-Hyun;Lee, Sang-Yong
    • Research in Plant Disease
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    • v.14 no.1
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    • pp.71-75
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    • 2008
  • An isolate of Peanut stunt virus (PSV), named as Tr-PSV, was isolated from white clover (Trifolium repens L) showing mosaic symptom. Tr-PSV systemically infected all plants tested in the Nicotiana spp. and induced local lesions on inoculated leaves of Chenopodium amaranticolor. However, Tr-PSV induced typical mosaic symptoms as ER-PSV on Vigna unguiculata 5 to 6 days after inoculation, while Fny-CMV used as a control virus of Cucumovirus produced local lesions on inoculated leaves. In dsRNA analysis, Tr-PSV consisted of four dsRNAs, but satellite RNA was not detected. The cDNA of coat protein gene of Tr-PSV was amplified by RT-PCR using a Cucumovirus-specific single pair primers that designed to amplify a DNA fragment of approximately 950 bp. By restriction mapping analysis using RFLP of the RT-PCR products and by serological properties of gel diffusion test, Tr-PSV belongs to a typical member of PSV subgroup I. This is the first report on the occurrence of PSV in white clover in Korea.

Type-I Hypersensitivity to Malassezia pachydermatis Extracts in Healthy Dogs and Dogs with Malasseza Otitis Externa (정상개와 Malassezia 외이염을 가진 개에 있어서 Malassezia pachydermatis 추출물의 즉시형 과민반응)

  • Kim, Young-Sub;Lee, Keun-Woo;Oh, Tae-Ho
    • Journal of Veterinary Clinics
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    • v.24 no.2
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    • pp.88-93
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    • 2007
  • The purpose of the study reported here was to test the hypotheses that clinically healthy dogs will not manifest immediate hypersensitivity responses to intradermal injection of Malassezia pachydermatis extracts but that affected dogs with Malassesia otitis will manifest such hypersensitivity. Wd desired to identify approximate molecular mass of any allergenic components of the yeast by use of sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The protein profile of Malassezia pachydermatis extracts showed between 16 and 110 kDa. Especially, the intensity was strongest between 25 and 80 kDa. Mean wheal diameters in the affected groups of 20, 2, 0.2, and $0.02{\mu}g/ml$ were $13.36{\pm}0.67,\;5.33{\pm}0.67,\;5.47{\pm}0.82,\;and\;5.07{\pm}0.64$, respectively. Mean wheal thickness in the affected groups of 20, 2, 0.2, and $0.02{\mu}g/ml$ was $6.44{\pm}0.40,\;3.86{\pm}0.35,\;2.64{\pm}0.36,\;and\;2.60{\pm}0.44$, respectively. The difference of wheal diameters and thickness between healthy and affected groups was significant (p<0.05). In conclusion, the observations confirm that Malassezia pachydermatis-derived antigens may induce an immediate wheal response when intradermal injected in dogs. It seems reasonable to suggest that hypersensitivity to yeast may contribute to the development of clinical signs in dogs with immediate skin test reactivity, especially in dogs with Malassezia otitis extema.

Immuno-stimulating Polysaccharides from the Fruiting Bodies of Fomitella fraxinea (II) -Isolation and characterization of hot-water extracted polysaccharides- (Fomitella fraxinea로부터 분리한 면역활성 다당류 (II) -열수추출 다당류의 분리 및 특성 -)

  • Cho, Soo-Muk;Lee, Jae-Hoon;Han, Sang-Bae;Kim, Hwan-Mook;Yu, Seung-Hun;Yoo, Ick-Dong
    • The Korean Journal of Mycology
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    • v.23 no.4 s.75
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    • pp.340-347
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    • 1995
  • Polysaccharide FHW was extracted from the fruiting bodies of Fomitella fraxinea with hot-water treatment and then fractionated into FHW-I and FHW-II on DEAE-Cellulose chromatography. FHW-I and FCW-II were further purified into FHW-Ia and Ib, FHW-IIa and IIb on gel permeation chromatography, respectively. A small amount of uronic acid was detected and glucose, galactose, fucose, and mannose were found to be main sugars in the polysaccharides. Protein was detected in FHW-Ia, FHW-IIa, and FHW-IIb, but not in FHW-Ib. FHW-Ia was identified to be a fuco-gluco-mannogalactan with molecular weight of 19,000 and FHW-Ib was a gluco-fuco-mannogalactan of 15,000. FHW-IIa and FHW-IIb were galacto-mannoglucan and their molecular weights were estimated to be 31,000 and 9,000, respectively. Both FHW-Ib and FHW-IIb did not show an absorption band characteristic of the ${\beta}-glycosidic$ linkage in IR spectra. FHW-IIb showed a strong immuno-stimulating activity but the other three polysaccharides showed a weak activity.

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