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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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Utilization Pattern of Complementary Therapy in Hypertension, Diabetes and Chronic Arthritis Patients Visited to Local Health Center (일개 보건소를 방문하는 고혈압, 당뇨 및 관절염환자의 보완요법 이용실태)

  • Park, Ae-Ju;Park, Jae-Yong;Han, Chang-Hyun
    • Journal of agricultural medicine and community health
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    • v.28 no.2
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    • pp.107-122
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    • 2003
  • Objectives: The objective of this study is to investigate the use rate and some aspect of complementary therapies used by patients with chronic illness(hypertension, diabetic mellitus and chronic arthritis). Methods: 600 patients visiting the health center for one month(Jan. 2001) were interviewed on their complementary therapies used by the subjects for the previous year. Results: About fourteen-eight percent of the respondents used therapies; 35% of patients with hypertension, 44.6% of patients with diabetic mellitus and 62.9% of patients with chronic arthritis, which shows the highest rate among patients with three chronic disease. The use rate of complementary therapies indicates few meaningful differences according to the general characteristics of the interviewees. Hypertension patients used herb medication(31.0%) acupuncture(29.6%) and most of all the other therapies. Diabetic patients used dietary therapy(57.5%) and herb medication(35.1%). Chronic arthritis patients used acupuncture(85%) and herb medication(34.7%). 36.8% of all the patients who used complementary therapies tried more than two therapies. 18.3% of hypertension patients, 24.1% of diabetic patients and 55.9% of chronic arthritis patients used more than two therapies. Acupuncture(47%) was used most frequently, followed by herb medications(26.3%), health assistance utensils(21.8%). oriental therapy(21.8%), physical therapy(9.5%), health assistance food(8.4%), herb(7.7%), Korea hand acupuncture(3.2%), abdomen respiration(1.1%), and pore therapy(0.7%) Oriental clinic was visited most frequently(42.8%), which was used to cure diseases(61.8%), and to relieve symptoms(26.0%). (p<0.001) The cost spent on complementary therapies last year was 90,000 won(40.3%) and there are some cases of more than 500,000 won(31.2%). Most of the patients(56.1%) were satisfied with the complementary therapies, with 6% of them having side effects. 74% of the patients used complementary therapies answered that they would continue them and 56.1% of them also answered that they would continue them and 56.1% of them also answered that they would advise other patients to do them. Advantages(compared with those of orthodox medical treatment) are psychological comfort(28.1%), body protection(26.0%), effectiveness(20.0%). 34% of the patients using complementary therapies wanted to have informational orientation on complementary therapies. These findings reveal that a considerable number of patients with chronic illness(47.5%) tried a variety of complementary therapies. Though 6% of the patients using therapies had side effects, most of the subjects seemed satisfied with them and they are supposed to continue them. Conclusions: In conclusion, health center personnels and medical doctors should pay more attention to the complementary therapies used by patients with chronic illness. They also have to try their best to advise more scientific and informative complementary programs with less side effects and more help to improve their conditions.

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Studies on Sclerotium rolfsii Sacc. isolated from Magnolia kobus DC. in Korea (목련(Magnolia kobus DC.)에서 분리한 흰비단병균(Sclerotium rolfsii Sacc.)에 관한 연구)

  • Kim Kichung
    • Korean journal of applied entomology
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    • v.13 no.3 s.20
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    • pp.105-133
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    • 1974
  • The present study is an attempt to solve the basic problems involved in the control of the Sclerotium disease. The biologic stranis of Sclerotium rolfsii Sacc., pathogen of Sclerotium disease of Magnolia kobus, were differentiated, and the effects of vitamins, various nitrogen and carbon sources on its mycelial growth and sclerotial production have been investigated. In addition the relationship between the cultural filtrate of Penicillium sp. and the growth of Sclerotium rolfsii, the tolerance of its mycelia or sclerotia to moist heat or drought and to Benlate (methyl-(butylcarbamoy 1)-2-benzimidazole carbamate), Tachigaren (3-hydroxy-5-methylisoxazole) and other chemicals were also clarified. The results are summarizee as follows: 1. There were two biologic strains, Type-l and Type-2 among isolates. They differed from each other in the mode of growth and colonial appearance on the media, aversion phenomenon and in their pathogenicity. These two types had similar pathogenicity to the Magnolia kobus and Robinia pseudoacasia, but behaved somewhat differently to the soybaen and cucumber, the Type-l being more virulent. 2. Except potassium nitrite, sodium nitrite and glycine, all of the 12 nitrogen sources tested were utilized for the mycelial growth and sclerotial production of this fungus when 10r/l of thiamine hydrochloride was added in the culture solution. Considering the forms of nitrogen, ammonium nitrogen was more available than nitrate nitrogen for the growth of mycelia, but nitrate nitrogen was better for sclerotia formation. Organic nitrogen showed different availabilities according to compounds used. While nitrite nitrogen was unavailable for both mycelial growth and sclerotial formation whether thiamine hydrochlioride was added or not. 3. Seven kinds of carbon sources examined were not effective in general, as long as thiamine hydrochloride was not added. When thiamine hydrochloride was added, glucose and saccharose exhibited mycelial growth, while rnaltose and soluble starch gave lesser, and xylose, lactose, and glycine showed no effect at all,. In the sclerotial production, all the tested carbon sources, except lactose, were effective, and glucose, maltose, saccharose, and soluble starch gave better results. 4. At the same level of nitrogen, the amount of mycelial growth increased as more carbon Sources were applied but decreased with the increase of nitrogen above 0.5g/1. The amount of sclerotial production decreased wi th the increase of carbon sources. 5. Sclerotium rolfsii was thiamine-defficient and required thiamine 20r/l for maximun growth of mycelia. At a higher concentration of more than 20r/l, however, mycelial growth decreased as the concentration increased, and was inhibited at l50r/l to such a degree of thiamine-free. 6. The effect of the nitrogen sources on the mycelial growth under the presence of thiamine were recognized in the decreasing order of $NH_4NO_3,\;(NH_4)_2SO_4,\;asparagine,\;KNO_3$, and their effects on the sclerotial production in the order of $KNO_3,\;NH_4NO_3,\;asparagine,\;(NH_4)_2SO_4$. The optimum concentration of thiamine was about 12r/l in $KNO_3$ and about 16r/l in asparagine for the growth of mycelia; about 8r/l in $KNO_3$ and $NH_4NO_3$, and 16r/l in asparagine for the production of sclerotia. 7. After the fungus started to grow, the pH value of cultural filtrate rapidly dropped to about 3.5. Hereafter, its rate slowed down as the growth amount increased and did not depreciated below pH2.2. 8. The role of thiamine in the growth of the organism was vital. If thiamine was not added, the combination of biotin, pyridoxine, and inositol did not show any effects on the growth of the organism at all. Equivalent or better mycelial growth was recognized in the combination of thiamine+pyridoxine, thiamine+inositol, thiamine+biotin+pyridoxine, and thiamine+biotin+pyridoxine+inositol, as compared with thiamine alone. In the combinations of thiamine+biotin and thiamine+biotin+inositol, mycelial growth was inhibited. Sclerotial production in dry weight increased more in these combinations than in the medium of thiamine alone. 9. The stimulating effects of the Penicillium cultural filtrate on the mycelial growth was noticed. It increased linearly with the increase of filtrate concentration up to 6-15 ml/50ml basal medium solution. 10. $NH_4NO_3$. as a nitrogen source for mycelial growth was more effective than asparasine regardless of the concentration of cultural filtrate. 11. In the series of fractionations of the cultural filtrate, mycelial growth occured in unvolatile, ether insoluble cation-adsorbed or anion-unadsorbed substance fractions among the fractions of volatile, unvolatile acids, ether soluble organic acids, ether insoluble, cation-adsorbed, cation-unadsorbed, anion-adsorbed and anion-unadsorbed. and anion-un-adsorbed substance tested. Sclerotia were produced only in cation-adsorbed fraction. 12. According to the above results, it was assumed that substances for the mycelial growth and sclerotial formation and inhibitor of sclerotial formation were include::! in cultural filtrate and they were quite different from each other. I was further assumed that the former two substances are un volatile, ether insotuble, and adsorbed to cation-exchange resin, but not adsorbed to anion, whereas the latter is unvolatile, ether insoluble, and not adsorbed to cation or anion-exchange resin. 13. Seven amino acids-aspartic acid, cystine, glysine, histidine, Iycine, tyrosine and dinitroaniline-were detected in the fractions adsorbed to cation-exchange resin by applying the paper chromatography improved with DNP-amino acids. 14. Mycelial growth or sclerotial production was not stimulated significantly by separate or combined application of glutamic acid, aspartic acid, cystine, histidine, and glysine. Tyrosine gave the stimulating effect when applied .alone and when combined with other amino acids in some cases. 15. The tolerance of sclerotia to moist heat varied according to their water content, that was, the dried sclerotia are more tolerant than wet ones. The sclerotia harvested directly from the media, both Type-1 and Type-2, lost viability within 5 minutes at $52^{\circ}C$. Sclerotia dried for 155 days at$26^{\circ}C$ had more tolerance: sclerotia of Type-l were killed in 15 mins. at $52^{\circ}C$ and in 5 mins. at $57^{\circ}C$, and sclerotia of Type-2 were killed in 10 mins. both at $52^{\circ}C$ or $57^{\circ}C$. 16. Cultural sclerotia of both strains maintained good germinability for 132 days at$26^{\circ}C$. Natural sclerotia of them stored for 283 days under air dry condition still had good germinability, even for 443 days: type-l and type-2 maintained $20\%$ and $26.9\%$ germinability, respectively. 17. The tolerance to low temperature increased in the order of mycelia, felts and sclerotia. Mycelia completely lost the ability to grow within 1 week at $7-8^{\circ}C$> below zero, while mycelial felts still maintained the viability after .3 weeks at $7-20^{\circ}C$ below zero, and sclerotia were even more tolerant. 18. Sclerotia of type-l and type-2 were killed when dipped into the $0.05\%$ solution of mercury chloride for 180 mins. and 240 mins. respectively: and in the $0.1\%$ solution, Type-l for 60 mins. and Type-2 for 30 mins. In the $0.125\%$ uspulun solution, Type-l sclerotia were killed in 180 mins., and those of Type-2 were killed for 90 mins. in the$0.125\%$solution. Dipping into the $5\%$ copper sulphate solution or $0.2\%$ solution of Ceresan lime or Mercron for 240 mins. failed to kill sclerotia of either Type-l or Type-2. 19. Inhibitory effect on mycelial growth of Benlate or Tachi-garen in the liquid culture increased as the concentration increased. 6 days after application, obvious inhibitory effects were found in all treatments except Benlate 0.5ppm; but after 12 days, distingushed diflerences were shown among the different concentrations. As compared with the control, mycelial growth was inhibited by $66\%$ at 0.5ppm and by $92\%$ at 2.0ppm of Benlate, and by$54\%$ at 1ppm and about $77\%$ at 1.5ppm or 2.0ppm of Tachigaren. The mycelial growth was inhibited completely at 500ppm of both fungicides, and the formation of sclerotia was checked at 1,000ppm of Benlate ant at 500ppm or 1,000ppm of Tachigaren. 20. Consumptions of glucose or ammonium nitrogen in the culture solution usually increased with the increment of mycelial growth, but when Benlate or Tachigaren were applied, consumptions of glucose or ammonium nitrogen were inhibited with the increment of concentration of the fungicides. At the low concentrations of Benlate (0.5ppm or 1ppm), however, ammonium nitrogen consumption was higher than that of the ontrol. 21. The amount of mycelia produced by consuming 1mg of glucose or ammonium nitrogen in the culture solution was lowered markedly by Benlate or Tachigaren. Such effects were the severest on the third day after their treatment in all concentrations, and then gradually recovered with the progress of time. 22. In the sand culture, mycelial growth was not inhibited. It was indirectly estimated by the amount of $CO_2$ evolved at any concentrations, except in the Tachigaren 100mg/g sand in which mycelial growth was inhibited significantly. Sclerotial production was completely depressed in the 10mg/g sand of Benlate or Tachigaren. 23. There was no visible inhibitory effect on the germination of sclerotia when the sclerotia were dipped in the solution 0.1, 1.0, 100, 1.000ppm of Benlate or Tachigaren for 10 minutes or even 20 minutes.

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Effect of Mixed Sowing Ratios Between Whole Crop Barley with Hooded Type and Forage Pea on the Forage Yield and Quality (삼차망 청보리와 사료용 완두의 혼파재배가 수량 및 사료가치에 미치는 영향)

  • Ju, Jung-Il;Park, Jong-Min;Lee, Jung-Jun;Kim, Chang-Ho;Koo, Han-Mo;Oh, Tae-Seok;Lee, Hyo-Won
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.29 no.3
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    • pp.171-178
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    • 2009
  • The study was conducted to clarify the mixed seeding rate of whole crop barley with hood type and forage pea for using of forage crops and to compare the forage yield and quality. At a mixed seeding rate between the whole crop barley (WCB) and forage pea, The heading date and plant height of WCB were not a difference according to mixed seeding rate of forage pea. The tillers of the WCB were a decrease and plant of the forage pea were a increase according to increased seeding rate of forage pea. The lodging index of the WCB was a appearance with distribution of $0{\sim}3$, The lodging index of WCB with a 20kg/10a seeding rate of a only WCB without seeding of the forage pea was 3. The overwintering rate of forage pea was a appearance more than 90% at all treatment. The plant height of forage pea was a increase according to increased seeding rate of forage pea at 14 kg/10a and 20 kg/10a plots of WCB. At a mixed seeding between the WCB and forage pea, The fresh weight was a increase according to increased seeding rate of forage pea and was a appearance more than 3,000 kg at all treatment plot. But the dry matter weight was decrease according to increased seeding rates of forage pea. The dry matter weight of 20 kg/10a seeding rate of a only WCB without seeding of the forage pea showed the most amount with 1,266 kg. The crude protein (CP) content was a tendency to increase according to increased seeding rates of forage pea. But, the relative feed value (RFV) was a tendency to decrease according to increased seeding rate of forage pea. The highest RFV was 183.8 at 14 kg/10a seeding rate of a only WCB without seeding of the forage pea. The acid detergent fiber (ADF) and neutral detergent fiber (NDF) were a increase according to increased seeding rate of forage pea at 14 kg/10a and 20 kg/10a plots of WCB. The highest content of ADF and NDF were 23.9% and 46.3% at mixed seeding rate of 20 kg/10a of WCB with 10 kg/10a of forage pea, respectively. The highest sum of standardized score by fresh weight, dry matter weight, CP, ADF, NDF and RFV was 2.309 at mixed seeding rate of 20 kg/10a of WCB with 7.5 kg/10a of forage pea. The optimum mixed seeding rate was a considered judgment in the order of mixed seeding rate of 20 kg/10a of WCB with 7.5 kg/10a of forage pea, mixed seeding rate of 20 kg/10a of WCB with 5.0 kg/10a of forage pea.

A Survey on the Break-down and Repair of the Power Tillers in Korea (동력경운기(動力耕耘機) 이용실태(利用實態) 조사분석(調査分析)(II) -고장(故障) 및 수리(修理)에 관(關)하여-)

  • Hong, Jong Ho;Lee, Chai Shik
    • Journal of Biosystems Engineering
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    • v.6 no.1
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    • pp.28-38
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    • 1981
  • A survey has been conducted to investigate the presents of breaks down and repair of power tiller for efficient use. Eight provinces were covered for this study. The results are summarized as follows. A. Frequency of breaks down. 1) Power tiller was breaken down 9.05 times a year and it represents a break down every 39.1 hours of use. High frequency of breaks down was found from the fuel and ignition system. For only these system, the number of breaks down were 2.02 and it represents 23.3% among total breaks down. It was followed by attachments, cylinder system, and traction device. 2) For the power tiller which was more than six years old, breaks down accured 37.7 hours of use and every 38.6 hours for the power tiller which was purchased in less than 2 years. 3) For the kerosene engine power tiller, breaks down occured every 36.8 hours of use, which is a higher value compared with diesel engine power tiller which break down every 42.8 hours of use. The 8HP kerosene engine power tiller showed higher frequency of break down compared with any other horse power tiller. 4) In October, the lowest frequency of break down was found with the value of once for every 51.5 hours of use, and it was followed by the frequency of break down in June. The more hours of use, the less breaks down was found. E. Repair place 1) 45.3% among total breaks down of power tiller was repaired by the owner, and 54.7% was repaired at repair shop. More power tiller were repaired at repair shop than by owner of power tiller. 2) The older the power tiller is, the higher percentage of repairing at the repair shop was found compared with the repairing by the owner. 3) Higher percentage of repairing by the owner was found for the diesel engine power tiller compared with the kerosene engine power tiller. It was 10 HP power tiller for the kerosene power tiller and 8 HP for the diesel engine power tiller. 4) 66.7% among total breaks down of steering device was repaired by the owner. It was the highest value compared with the percentage of repairing of any other parts of power tiller. The lowest percentage of repairing by owner was found for the attachments to the power tiller with the value of 26.5%. C. Cause of break down 1) Among the total breaks down of power tiller, 57.2% is caused by the old parts of power tiller with the value of 5.18 times break down a year and 34.7% was caused by the poor maintenance and over loading. 2) For the power tiller which was purchased in less than two years, more breaks down were caused by poor maintenance in comparison to the old parts of power tiller. 3) For the both 8-10 HP kerosene and diesel engine power tiller, the aspects of breaks down was almost the same. But for the 5 HP power tiller, more breaks down was caused by over loading in comparison to the old parts of power tiller. 4) For the cylinder system and traction device, most of the breaks down was caused by the old parts and for the fuel and ignition system, breaks down was caused mainly by the poor maintenance. D. Repair Cost 1) For each power tiller, repair cost was 34,509 won a year and it was 97 won for one hoar operation. 2) Repair cost of kerosene engine power tiller was 40,697 won a year, and it use 28,320 won for a diesel engine power tiller. 3) Average repair cost for one hour operation of kerosene engine power tiller was 103 won, and 86 won for a diesel engine power tiller. No differences were found between the horse power of engines. 4) Annual repair cost of cylinder system was 13,036 won which is the highest one compared with the repair cost of any other parts 362 won a year was required to repair the steering device, and it was the least among repair cost of parts. 5) Average cost for repairing the power tiller one time was 3,183 won. It was 10,598 won for a cylinder system and 1,006 won for a steering device of power tiller. E. Time requirement for repairing by owner. 1) Average time requirements for repairing the break down of a power tiller by owner himself was 8.36 hours, power tiller could not be used for operation for 93.58 hours a year due to the break down. 2) 21.3 hours were required for repairing by owner himself the break down of a power tiller which was more than 6 years old. This value is the highest one compared with the repairing time of power tiller which were purchased in different years. Due to the break down of the power tiller, it could not be used for operation annually 127.13 hours. 3) 10.66 hours were required for repairing by the owner himself a break down of a diesel engine power tiller and 6.48 hours for kerosene engine power tiller could not be used annually 99.14 hours for operation due to the break down and it was 88.67 hour for the diesel engine power tiller. 4) For both diesel and kerosene engine power tiller 8 HP power tiller required the least time for repairing by owner himself a break down compared with any other horse power tiller. It was 2.78 hours for kerosene engine power tiller and 8.25 hours fur diesel engine power tiller. 5) For the cylinder system of power tiller 32.02 hours were required for repairing a break down by the owner himself. Power tiller could not be used 39.30 hours a year due to the break down of the cylinder system.

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