• Title/Summary/Keyword: 분자량분포

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Analysis of Clonorchis sinensis antigens and diagnosis of clonorchiasis using monoclonal antibodies (단세포군 항체를 이용한 간흡충 항원의 분석 및 간흡충증의 진단)

  • Yong, Tae-Sun;Im, Gyeong-Il;Jeong, Pyeong-Rim
    • Parasites, Hosts and Diseases
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    • v.29 no.3
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    • pp.293-310
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    • 1991
  • Clonorchis sinensis is a common parasite of man in Korea. Researches on the specific antigens of C. sinensis would be valuable not only because those elucidate the molecular characteristics of this fluke but also because it is applicable to immunodiagnosis. Although many monoclonal antibodies have been used in the field of parasite immunology, few articles on monoclonal antibodies against C. sinensis have been published so far. The aim of this study was to analyse C. sinensis antigens recognized by monoclonal antibodies, and to set up ELISA-inhibition test using C. sinensis specific monoclonal antibodies for improved specificity of immunodiagnostic tests. By fusion between spleen cells of the mice immunized with C. sinensis water-soluble crude adult worm antigens and plasmacytoma cells of mouse origin, 29 hybridoma clones secreting anti-C. sinensis monoclonal antibodies were made, and 8 clones among those were found specific. After cell cloning, isotypes of 6 selected specific monoclonal anti- bodies were determined to be IgGl, IgG2b and IgA. Four exposed antigenic determinants of natural infection were recognized by different specific monoclonal antibodies. By enzyme-immunoelectrotransfer blot, 10 KD, 34 KD antigenic determinants were found to be reacted with CsHyb 0714-20, CsHyb 0605-10 monoclonal antibodies, respectively, The antigenic determinant recognized by CsHyb 0714-20 monoclonal antibody was revealed to be located at the surface and parenchyme of a parasite by indirect immunoauorescent antibody technique, and those reacted with CsHyb 0605-10, CsHyb 0714-25 monoclonal antibodies were found at the parenchyme and intestine. The antigenic determinant reacted with CsHyb 0605-23 monoclonal antibody was found mainly around the uterine eggs. Four antigenic determinants recognized by specific monoclonal antibodies were all found to be present in the early eluted fractions of C. sinensis antigens separated by Sephadex G-200 gel filtration. By conventional ELISA, 75% of clonorchiasis cases were found positive, but 7.1% of normal controls and 37.5% of paragonimiasis cases showed false positives. However, by ELISA-inhibition test using C. sinensis specific monoclonal antibody (CsHyb 0605-23), 77.1% of clonorchiasis cases were found positive, and there were no false positives in normal controls or paragonimiasis cases, indicating 100% specificity. The ELISA- inhibition test using monoclonal antibodies was found to have same sensitivity and definitely high specificity in comparison with conventional ELISA for serodiagnosis of human clonorchiasis.

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Hydrolysis of Fish Protein Concentration in an Ultrafiltration Membrane Reactor (한외여과막 반응기를 이용한 FPC의 가수분해)

  • 최정호;변희국;김세권
    • Membrane Journal
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    • v.10 no.2
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    • pp.83-91
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    • 2000
  • In order to improve functional properties, enzymatic hydrolysis of FPC (fish protein concentration) was achieved in ultrafiltration membrane reactor (MWCO 5,000). First, insoluble FPC was hydrolyzed by pepsin in batch reactor to decrease the fouling in ultrafiltration membrane reactor, and second hydrolysis was achieved by pronase E in ultrafiltration membrane reactor The optimum operating conditions in batch reactor using pepsin were at temperature 45$^{\circ}C$, pH 2.0 and the ratio of substrate to pepsin, 150 (w/w) After operating for 5hrs under optimum conditions, 89% of total amount of initial FPC was hydrolyzed. The rate constants, $K_{m}$ and V$_{max}$, were 1.25% and 0.89 mg/$m\ell$/min, respectively, and substrate inhibition was occured above 1.5%. The ultrafiltration membrane reactor was operated with recycling rate of 474 $m\ell$/min and transmembrane pressure of 15 psi. The permeate flux was increased by temperature, transmembrane pressure, but the permeate flux was fixed by pH. The optimum ratio of substrate to pronase E was 200(w/w) and the productivity of ultrafiltration membarane reactor was 702 mg/mg -enzyme, that of batch reactor was 51mg/mg-enzyme. Molecular weight distributions tot first and second hydrolysates were from 2,500 Da to 20,000 Da and from 700 Da to 10,000 Da, respectivelyly.

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Characterization of Flacherie Virus of the Silkworm, Bombyx mori, isolated in Korea (한국산 연화병 바이러스의 분리 및 성상)

  • 김근영;강숙우
    • Journal of Sericultural and Entomological Science
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    • v.33 no.1
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    • pp.13-20
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    • 1991
  • The flacheric virus(FV) was isolated from the diseased larvae with flaccidity symptoms of the silkworm, Bombyx mori, which were collected in Hyangnam, Kyunggi Province and in Youcheon, Kyungpook Province. The properties of the virus were investigated and compared with the Japanese FV in morphology, size, nucleic acid and structural proteins. The Hyangnam isolate had a diameter of 27$\pm$1, 7nm with speherical shape and contained RNA of which the electrophoretic pattern was same as that of the Japanese FV. The virus had four sturctural proteins and their molecular weight were estimated as 35, 000, 33, 000, 31, 000, and 11, 400 daltons, respectively. The Youcheon isolate had two different sizeds in diameter of 27$\pm$1.7nm and 21$\pm$0.8nm. The antigenicity of the Hyangnam isolate was proved to be identical to that of Japanese FV, whereas the antisera against the Youcheon isolate (mixed with two different sizes) reacted with Japanese FV and densonucleosis virus type 1, respectively. From these characteristic of the isolated viruses, it was concluded that the Hyangnam isolate was identical to the Japanese FV and the Youncheon isolate contained the same viruses as the Japanese FV and DNVI.

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Comparative Studies on the Enzymatic Properties of two Trypsin-like Enzymes from Menhaden, Brevoortia tyranus (혈합육어 멘헤이든의 장기조직분포Trypsin-유사효소에 관한 비교효소학적 연구)

  • PYEUN Jae-Hyeung;KIM Hyeung-Rak;GODBER J. S.
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.23 no.1
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    • pp.12-24
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    • 1990
  • Two trypsin-like enzymes, designated trypsin A and 3, purified from the intestine of menhaden by $(NH_4)_2SO_4$ fractionation, Benzamidine-Sepharose 6B affinity chromatography, DEAE-Sephacel ion exchange chromatography and Sephadex G-75 gel filtration chromatography. The two trypsins were subjected to compare the enzymatic properties of the trypsin-like enzymes from the other dark fleshed fishes. Both trypsins catalysed the hydrolysis of N$\alpha$-benzoyl-DL-arginine-p-nitroanilide and they were remarkably inhibited by several well known trypsin-inhibitors, tosyllysyl chloromethyl ketone, soybean trypsin inhibitor, be-nzamidine, leupeptin and antipain, etc. Therefore, it was ascertained that the two enzymes are serine-type trypsins. The molecular weights of these enzymes were about 25,000 and 26,200, respectively, ;Is determined by SDS-PAG electrophoresis and by Sephadex G-100 gel filtration, and the molecular weights of these two enzymes are somewhat fewer than those from the other dark fleshed fishes. Both enzymes had less basic amino acids such as arginine and Iysine, whereas they had slightly high contents of neutral amino acids, glycine, alanine and tryptophane. The enzymes showed a pH optimum of $8\~11$ at $60^{\circ}C$ against the $N\alpha$-benzoyl-DL-argi-nine-p-nitroanilide substrate and they were quite unstable above $40^{\circ}C$ and under the atidic pH region. The Km constant of the two enzymes against the $N\alpha$-benzoyl-DL-arginine-p-nitroanilide was $1.4\times10^{-4}M$ for trypsin A and $4.3\times10^{-5}M$ for trypsin B, respectively.

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Analysis on Pesticide Residue in Medicinal Dried Fruits (약용건조과실류의 잔류농약분석)

  • Lee, Kyeong-Ah;Kang, Sung-Tae;Kim, Ouk-Hee;Park, Sung-Kyu;Ha, Kwang-Tae;Choi, Young-Hee;Jo, Han-Bin;Choi, Byung-Hymn
    • The Korean Journal of Pesticide Science
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    • v.14 no.3
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    • pp.209-218
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    • 2010
  • There experiment were carried out to analyze for pesticide residues in 4 kinds of dried fruits collected in northern area of Seoul in 2007~2009. Total of 213 samples were analyzed. According to the results, 20 kinds of residual pesticides were detected. Residual pesticides were detected in 66 samples (31.0%) and 8 samples (3.8%) exceeded maximum residue limits (MRL). Cypermethrin, fenvalerate, endosulfan, chlorpyrifos, bifenthrin were detected frequently. The Pesticide types which were detected in dried furits were revealed in order of pyrethroid > organophosphate > organochloride > dicarboximide. Detection rates of dried fruits were dried chinese matrimony vein (53.0%), dried jujube (34.9%), dried rubi fructus (7.6%) and dried maximowiczia chinensis (4.6%). Based on these results, we investigated the risk assessment from amount of residual pesticide in dried jujube. Range of %ADI were 0.0001~0.0081%, but the value has not effected on human health.

Changes in Pectic Substances of Mature-Green Mume (Prunus mume Sieb. et Zucc) Fruit as Influenced by the Thickness of Packaging Film during Storage (포장필름의 두께가 청매실의 저장중 펙틴질의 변화에 미치는 영향)

  • Cha, Hwan-Soo;Chung, Myong-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.4
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    • pp.621-628
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    • 2002
  • Effect of the thickness of packaging film on the ripening of mature-green mume (Prunus mume Sieb. et Zucc) fruit was investigated by measuring physicochemical changes of the fruit during storage. Fruits were packaged using low density polyethylene (LDPE) films with thicknesses of 20, 30, and 40$\mu$m and stored at the room temperature. The physicochemical properties such as contents of various pectic substances, molecular weight distribution of soluble pectic substances, and surface image of the fruit were determined during storage of 8 days. In general, regardless of the thickness of the films applied, a content of water-soluble pectin (WSP) in the fruit was increased during storage, but both contents of HCI- soluble pectin (HSP), and Ca and Mg in total alcohol-insoluble solids were decreased. Sephacryl S-300 gel filtration study revealed that fractions of high molecular weight HSP and WSP decomposed into lower molecular weight pectins during storage. The scanning electronic microscope also identified a significant structural change of the fruit skin over the storage time. It could be concluded from the results that fruits packaged with LDPE 30${\mu}{\textrm}{m}$ film maintained the highest physicochemical quality of green mume fruit during storage.

Synthesis and Characterization of Biodegradable MethoxyPoly(ethylene glycol)-Poly$(\varepsilon-caprolactone-co-L-lactide)$ Block Copolymers (메톡시폴리(에틸렌 글리콜)-폴리(카프로락톤-co-L-락타이드) 공중합체의 합성 및 특성 분석)

  • Hyun Hoon;Cho Young Ho;Jeong Sung Chan;Lee Bong;Kim Moon Suk;Khang Gilson;Lee Hai Bang
    • Polymer(Korea)
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    • v.30 no.1
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    • pp.28-34
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    • 2006
  • A series of methoxypoly(ethylene glycol) $(MPEG)-poly(\varepsilon-co-L-lactide)$ (PCLA) diblock copolymers were synthesized by ring-opening polymerization of a mixture of $\varepsilon-caprolactone$ and L-lactide with different ratios in the presence of $Sn(Oct)_2$. The characterization of MPEG-PCLA diblock copolymers were examined by $^1H-NMR$, GPC, DSC, and XRD. Kinetic study on ring-opening polymerization of monomer mixtures was carried out in various conditions such as a variation with polymerization time, amount of catalyst, and temperature. The highest conversion obtained in 1.2 ratic of initiator venn catalyst at $110\;^{\circ}C$. The biodegradable characterization of MPEG-PCLA diblock copolymers in aqueous solution was carried out by using GPC for $1\~14$ weeks. The biodegradability of MPEG-PCLA diblock copolymers increased as the L-lactide content of diblock copolymers increased. In conclusion, we confirmed the dependence of polymerization rate according to various conditions. In addition, we can control the biodegradability of MPEC-PCLA diblock copolymers by changing the ratio of PCL and PLA block segment.

Cloning of Pig Kidney cDNA Encoding an Angiotensin I Converting Enzyme (돼지 신장의 Angiotensin I Converting Enzyme cDNA 클로닝)

  • Yoon, Jang-Ho;Yoon, Joo-Ok;Hong, Kwang-Won
    • Applied Biological Chemistry
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    • v.49 no.4
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    • pp.293-297
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    • 2006
  • Angiotensin converting enzyme(ACE) is a zinc-containing dipeptidase widely distributed in mammalian tissues and is thought to play a significant role in blood pressure regulation by hydrolyzing angiotensin I to the potent vasoconstrictor, angiotensin II. Recently, the presence of ACE in pig ovary was reported and the ACE from pig kidney was isolated and characterized. However no nucleotide sequence of the ACE gene from pig is yet known. We report here the cloning of the ACE cDNA from pig kidney by using the reverse transcriptase-polymerase chain reaction. The complete amino acid sequence deduced from the cDNA contains 1309 residues with a molecular mass of 150 kDa, beginning with a signal peptide of 33 amino acids. Amino acid sequence analysis showed that pig kidney ACE is also probably anchored by a short transmembrane domain located near the C-terminus. This protein contains a tandem duplication of the two homologous amino acid peptidase domain. Each of these two domains bears a putative metal-binding site (His-Glu-Met-Gly-His) identified in mammalian somatic ACE. The alignment of pig ACE amino acid sequence with human, rabbit, and mouse reveals that both two domains have been highly conserved during evolution.

Immunological properties of the 30 kDa antigen of Toxoplasma gondii (단클론 항체를 이용하여 정제한 톡소포자충 30 kDa 항원의 면역학적 특성)

  • Lee, Yeong-Hwa;No, Tae-Jin;Sin, Dae-Hwan
    • Parasites, Hosts and Diseases
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    • v.35 no.1
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    • pp.55-62
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    • 1997
  • The molecular weight 30 kDa membrane protein of Toxoplusma Sondii (Toxoplasma 30 kDa) apparently conserved in most strains of T. gondii and sera of infected hosts. The present study aimed to elucidate Toxoplasmc 30 kDa as a useful diagnotic antigen for serodiagnisis of toxoplasmosis by ELISA and for induction of protective immunity. Murine spleen cells immunized with the membrane antigen of T. gondii were fused with mouse Sp2/0-Ag 14 myeloma cells. Out of 8 clones selected, five were IgG2b, the others belonged to IgG 1 and IgG2a. The 30 kDa antigen was distributed mainly on the surface membrane of tachyzoites by indirect fluorescence method. Murine peritoneal macrophages which were activated by 30 kDa antigen produced more amounts of NO2 compared with crude antigen-treated group, however there were no significant differences in toxoplamacidal activity between the two groups. Higher specificity of Toxoplosma 30 kDa antigen was recognized for serodiagnosis of toxoplasmosis than the crude antigen. From these results, ToxopLasmo 30 kDa antigen enhances the cytotoxic effect of macrophages as well as a more reliable means for the serodiagnosis of toxoplasmosis by ELISA. Key words: Toxoplosma gondii, 30 kDa antigen (p30), mouse, serodiagnosis, macrophage, cytotoxicity.

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Characterization of Korean Sweet Potato Starches: Physicochemical, Pasting, and Digestion Properties (국내 육종 고구마 전분의 이화학 호화 및 소화 특성)

  • Baek, Hye Rim;Kim, Ha Ram;Kim, Kyung Mi;Kim, Jin Sook;Han, Gui Jung;Moon, Tae Wha
    • Korean Journal of Food Science and Technology
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    • v.46 no.2
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    • pp.135-142
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    • 2014
  • Physicochemical, pasting, and digestion properties of sweet potato starches from 11 Korean cultivars were investigated. Starch granules were variably oval, round, polygonal, spherical, and bell-shaped, and of 10.2-15.3 ${\mu}m$ in mean particle diameter. Amylose contents varied from 12.3 to 17.4%. A similar chain length distribution of amylopectin was found in each of the cultivars. The portion of $B_3$ correlated with the degree of amylose leaching. Thermal properties determined by differential scanning calorimetry showed high values of gelatinization temperatures in Shinyulmi and Jeungmi starches, but a relatively low value in Daeyumi starch. All starches exhibited a Ca-type diffraction pattern. Differing patterns were observed in swelling factors, depending on temperature. The contents of rapidly digestible starch, slowly digestible starch, and resistant starch ranged from 9.6-17.4, 31.4-45.6, and 35.7-62.8%, respectively. In Rapid Visco Analyser profiles, differences were observed in pasting parameters such as pasting temperature, peak viscosity, final viscosity, and breakdown.