• Title/Summary/Keyword: Protein fractions

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Purification of antigenic proteins of Paragonimus westermani and their applicability to experimental cat paragonimiasis (폐(肺)디스토마(Paragonimus westermani) 감염(感染) 고양이 혈청(血淸)에 대(對)한 ELISA 항체가(抗體價)의 의의(意義))

  • Choi, Won-Young;Yoo, Jae-Eul;Nam, Ho-Woo;Choi, Hyung-Rak
    • Parasites, Hosts and Diseases
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    • v.24 no.2
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    • pp.177-186
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    • 1986
  • This study was designed to evaluate the partially purified antigens which were fractionated from crude extract of Paragonimus westermani and to monitor the enzyme-linked immunosorbent assay (ELISA) in experimental cat paragonimiasis during the course of infection as well as before and after chemotherapy. Crude extract of 6-month-old adult P. westermani was fractionated to 5 antigens by successive applications of ammonium sulfate precipitation, ion exchange chromatography and gel filtration. And the cats, 10 in each group, were infected with 60, 30, 15, and 5 metacercariae, then the half of each group was treated with praziquantel 2 times in one day of 100mg per kilogram of weight on 150 days after the infection. Sera were collected every 10 days. ELISA was performed with the concentration of $2{\mu}g/ml$ antigen, 100 times diluted sera and 1,000 times diluted alkaline phosphatase conjugated anti-cat IgG. The results were as follows: 1. Absorbance by ELISA with proteins precipitated by differential concentration of ammonium sulfate was the highest at $51{\sim}65%$ precipitate (PA2), followed by $0{\sim}50%$ precipitate (PAl), $66{\sim}80%$ precipitate (PA3), and $81{\sim}90%$ precipitate (PA4). Unprecipitated protein over 90% ammonium sulfate (PA5) showed the lowest antigenicity. 2. Fractionation of PA1, PA2, and PA3 through the DEAE-cellulose column did not differentiate the antigenic proteins. 3. By passing through the Sephadex G-200 column, PA1 and PA2 were fractionated to high molecular weight proteins and those of low molecular weight which showed high absorbance by ELISA (PA1-I, II and PA2-I, II). But PA3 was shown to have a fraction of high molecular weight proteins (PA3-I) which showed high antigenicity. 4. SDS-polyacrylamide gel electrophoresis of PA1-I, P A1-II, PA2-I, PA2-II, PA3-I, and crude extract was performed. Fraction PA1-I was composed of proteins which had the molecular weight of 270 kilodaltons(KD) to 196 KD; of them 220KD protein was major band. Fraction PA2-I was composed of $255{\sim}225\;KD$, and PA3-I, $255{\sim}240\;KD$, respectively. Fraction PA1-II and fraction PA2-II consisted of 30 KD proteins. 5. Absorbance by ELISA began to increase within $10{\sim}20$ days after the infection and reached the highest on $140{\sim}180$ days, then made plateau thereafter. 6. Absorbance by ELISA decreased after praziquantel treatment. In 60 metacercariae infection group, the absorbance had been decreasing, but remained within the positive range during observation period, while those of 30, 15, and 5 metacercariae infection groups turned to negative range. 7. Fraction PA1-II showed the highest antigenicity in ELISA, then fraction PA2-I, fraction PA1-I, fraction PA2-II, fraction PA3-I and crude extract followed. In early phase of infection, the absorbance of fraction PA1-II showed more rapid increase than those of the other fractions and it came to positive range at $20{\sim}30$ days after infection.

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Food Components of Wild and Cultured Fresh Water Fishes (천연 및 양식산 담수어의 식품성분)

  • KIM Kyung-Sam;LEE Eung-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.19 no.3
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    • pp.195-211
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    • 1986
  • The object of this study is to obtain fundamental data on cultured fishes produced in Korea to improve their food components. For this purpose, the food components of cultured fresh water fishes such as eel, Anguilla japonica, snakehead, Channa argus, and common carp, Cyprinus carpio, were investigated and compared with those of the wild ones. The results obtained are summarized as follows: 1. Common characteristics in the proximate composition were that wild fish was higher in crude protein content and lower in crude lipid content than those of cultured one. 2. Among the 9 kinds of minerals analyzed in all the samples, sodium, potassium, calcium and magnesium contents were absolutely predominant being more than $99.52\%$. These four elements in feedstuff also occupied $99.68{\sim}99.92%$ of total minerals. 3. The neutral lipids of wild and cultured eel, snakehead and common carp occupied $55.7{\sim}95.8%$ of lipid fractions, while the content of the phospholipids in snakehead was particularly higher than those of others. 4. The neutral lipids of wild and cultured eel, snakehead and common carp mainly consisted of triglycerides ($85{\sim}95%$), and a little quantity of diglycerides, monoglycerides, free sterol ester and hydrocarbon were also identified in the neutral lipid. 5. The phospolipids of eel and common carp were mainly occupied by phosphatidyl choline ($71.3{\sim}83.9%$), followed by phosphatidyl ethanolamine ($12.1{\sim}23.5%$) and phosphatidyl serine ($7.5{\sim}13.8%$). The phospholipids of snakhead consisted of phosphatidyl choline ($50.7{\sim}64.5%$), phosphatidyl ethanolamine ($28.0{\sim}35.5%$) and phosphatidyl serine ($7.5{\sim}13.8%$). Generally, phosphatidyl choline content was higher in wild fish than in cultured one, while phosphatidyl ethanolamine and phosphatidyl serine contents were higher in cultured one. 6. The major fatty acids in total lipid of wild eel, snakehead and common carp were $C_{16:0}\;and\;C_{20:5}$, while those in cultured ones were $C_{18:1},\;C_{18:2}\;and\;C_{22:6}$. The fatty acid composition of neutral lipids showed similar tendency to that of total lipid, and the main fatty acids in phospholipids of cultured fishes were $C_{18:1}\;and\;C_{18:2}$. In glycolipids, $C_{20:5}\;and\;C_{22:6}$ were higher in wild fishes, while $C_{18:2}$ were higher in cultured ones. 7. Total amino acids contents of wild and cultured eel were nearly the same, being $16.65\%$ ana $15.99\%$ respectively. The major amino acids of wild and cultured fish were glutamic acid, leucine, aspartic acid and lysine in order. In snakehead, the contents of aspartic acid and proline in cultured fish were higher than those in wild one, while the contents of glutamic acid, alanine, glycine were higher in the wild one. Total amino acid content of cultured common carp was $21.7\%$ compared with $17.08\%$ in wild one. The contents of glutamic acid, aspartic acid, glycine, proline and alanine occupied higher quantities in cultured common carp compared with those in wild one while the other amino acids revealed no significant difference. 8. Aspartic acid in free amino acids of cultured eel held $1.0\%$ of total free amino acids, while that in wild eel held $2.9\%$. Histidine, arginine and tyrosine content of cultured fish were two times higher than those of wild one. But free amino acid composition of samples seemed to be no marked differences according to cultured places. The contents of arginine, aspartic acid, glutamic acid, methionine and phenylalanine of snakehead ware higher in wild one than in cultured one, while the contents of lysine, histidine, glycine, and alanine ware higher in cultured one. In free amino acids content of wild common carp, histidine, glycine and lysine occupied $76.9\%$ of total free amino acids. Lysine, histidine, aspartic acid, alanine, valine and leucine were higher in wild one compared with those of cultured one, while glycine and tyrosine contents were higher in cultured fish.

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