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Development of Assay Methods for Enterotoxin of Escherichia coli Employing the Hybridoma Technology (잡종세포종기법을 이용한 대장균의 장독소 측정법 개발)

  • Kim, Moon-Kyo;Cho, Myung-Je;Park, Kyung-Hee;Lee, Woo-Kon;Kim, Yoon-Won;Choi, Myung-Sik;Park, Joong-Soo;Cha, Chang-Yong;Chang, Woo-Hyun;Chung, Hong-Keun
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.1
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    • pp.151-161
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    • 1986
  • In order to develop sensitive and sepcific assay methods for E. coli heat labile enterotoxin(LT) hybridoma cell lines secreting LT specific monoclonal antibody were obtained. LT was purified from cell lysate of E. coli O15H11. The steps included disruption of bacteria by French pressure, DEAE Sephacel ion exchange chromatography, Sephadex G200 gel filtration, and second DEAE Sephacel ion exchange chromatography, successively. Spleen cells from Balb/c mice immunized with the purified LT and $HGPRT^{(-)}$ plasmacytomas, $P3{\times}63Ag8.V653$ were mixed and fused by 50% (w/v) PEG. Hybrid cells were grown in 308 wells out of 360 wells, and 13 wells out of them secreted antibodies reacting to LT. Among these hybridoma cell 1G8-1D1 cell line was selected since it had produced high-titered monoclonal antibody continuously. By using culture supernatant and ascites from 1G8-1D1 cells the monoclonal antibody was characterized, and an assay system for detecting enterotoxigenic E. coli was established by double sandwich enzyme-linked immunosorbent assay (ELISA). The following results were obtained. 1. Antibody titers of culture supernatant and ascites from 1G8-1D1 hybridoma cells were 512, and 102, 400, respectively by GM1-ELISA and its immunoglobulin class was IgM. 2. The maximum absorption ratio of 1G8-1D1 cell culture supernatant to LT was 90% at $300\;{\mu}g/ml$ of LT concentration. LT concentration shown at 50% absorption ratio was $103.45{\mu}g$ and the absorption ratio was decreased with tile reduction of LT concentration. This result suggests that monoclonal antibody from 1G8-1D1 hybridoma cell bound with LT specifically. 3. The reactivities of 1G8-1D1 cell culture supernatant to LT and V. cholerae enterotoxin(CT) were 0.886 and 0.142(O.D. at 492nm) measured by the GM1-ELISA, indicating 1G8-1D1 monoclonal antibody reacted specifically with LT but not with CT. 4. The addition of 0.1ml of ascites to 0.6mg and 0.12mg of LT decreased the vascular permeability factor to 41% and 44% respectively, but it did not completely neutralize LT. 5. By double sandwich ELISA using monoclonal antibody, as little as 75ng of the purified LT per ml could be detected. 6. The results by assay of detecting LT in culture supernatants of 14 wild strains E. coli isolated from diarrhea patients by the double sandwich ELISA were almost the same level as those by reverse passive latex agglutination.

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A Study on Effects of Air-delivery Rate upon Drying Rough Rice with Unheated Air. (벼의 자연통풍건조에 있어서 통풍량이 건조에 미치는 영향에 관한 연구)

  • 이상우;정창주
    • Magazine of the Korean Society of Agricultural Engineers
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    • v.16 no.1
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    • pp.3293-3301
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    • 1974
  • An experimental work was conducted by using a laboratory-made model dryer to investigate the effect of the rate of natural forced-air on the drying rate of rough rice which was deposited in the deep-bed. The dryer consisted of 8 cylinderical containers with grain holding screen at their bottoms, each of which having 30cm in diameter and 15cm in height. The containers were sacked vertically with keeping them air-tight by using paper tape during dryer operation. Two separate layers of containers were operated in the same time to have two replications. The moisture contents of grains within each bins after predetermined period of dryer operation were determined indirectly by measuring the weight of the individual containers. The air-rates were maintained at 6 levels, or 5, 8, 10, 15, 18 and 20 millimenters of static head of water. The roomair conditions during dryer operation were maintained in the range of 10-l5$^{\circ}C$ in temperature and 40-60% in relative humidity. The results of the study are summarized as follows: 1. Drying characteristics of the grains in the bottom layers were approximately the same regardless of airdelivery rates, giving the average drying rate as about 0.35 percent per hour after 40-hour drying period, during which moisture content (w. b.) reduced from 24 percent to about 10 percent. 2. After about 40-hour drying period, the mean drying rates increased from 0.163 percent per hour to 0.263 percent per hour as air-flow rates increased from 5mm to 87.16mm of static head of water. In the same time, the moisture differences of grains between lower and upper layers varied from 12.7 percent at the air rate of 5mm of water head to 7.5 percent at the air-flow rate of 20mn of water head. Thus, the greater the air-flow rate was, the more overall improvement in drying performance was. Additionally, from the result of ineffectiveness of drying grain positioned at 70cm depth or above by the air rate of 5mm of static head of water it may be suggested in practical application that the height of grain deposit would be maintained adequately within the limits of air-rates that may be actually delivered. 3. Drying after layer-turning operation was continued for about 30 hours to test the effectiveness of reducing moisture differences in the thick layers. As a result of this layer-turning operation, moisture distribution through layers approached to narrow ranges, giving the moisture range as about 7 percent at air-flow rate of 5mm head of water, about 3 percent at 10mm head about 2 percent at 15mm head, and less than 1 percent at 20mm head. In addition, from the desirable results that drying rate was rapid in the lower layers and dully in the upper layers, layer-turning operation may be very effective in natural air drying with deep-layer grain deposit, especially when the forced air was kept in low rate. 4. Even though the high rate of air delivery is very desirable for deep-layer natural-air drying of rough rice, it can be happened that the required air delivery rate could not be attained because of limitation of power source available on farms. To give a guide line for the practical application, the power required to perform the drying with the specified air rate was analyzed for different sizes of drying bin and is given in Table (5). If a farmer selects a motor of which size is 1 or {{{{1 { 1} over {2 } }}}} H.P. and air-delivery rate which ranges from 8~10mm of head, the diameter of grain bin may be suggested to choose about 2.4m, also power tiller or other moderate size of prime motor may be recommended when the diameter of grain bin is about 5.0m or more for about 120cm grain deposit.

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Development of Transgenic NT Embryos Using Bovine Fetal Fibroblasts Transfected with hFSH Gene (hFSH 유전자가 도입된 소 태아섬유아세포를 이용한 형질 전환 복제 수정란의 발달)

  • Yang B.C.;Im G.S.;Kim D.H.;Min K.S.;Yoon D.H.;Park H.S.;Kim S.W.;Hwang I.S.;Seo J.S.;Seong H.H.;Yang B.S.
    • Journal of Embryo Transfer
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    • v.21 no.1
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    • pp.13-20
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    • 2006
  • The purpose of this study was to develope the transgenic cattle expressing hFSH into the urine using the nuclear transfer. To produce the interest gene in urine, the specific vector was ligated with hFSH gene undo. maUII promoter. The fetal fibroblast cells (KbFF) were isolated from a 45-day male fetus. The hFSH gene was co-transfected with pcDNA3 (neo) vector to KbFF cells by electroporation. The gene-transfected cells were cultured with G-418 selection medium for 2 weeks. Selected colonies were confirmed by PCR. For nuclear transfer, enucleated bovine oocytes were transferred with hFSH transfected or nontransfected fetal fibroblasts. The cleavage and blastocyst formation rates were significantly lower (p<0.05) in cloned embryos transfected with hFSH gene (68.7% and 15.7%) than in those non-transfected (67.6% and 24.5 %), respectively. Apoptosis analysis showed no difference between hFSH transfected and non-transfected blastocysts (p>0.05). The blastocysts were transfected to 77 (control 24, hFSH 53) recipient cows. Two calves were born (1.9%) following transfer with NT embryos transfected with hFSH gene, but they were confirmed not to be transgenic calves. This result shows that the hFSH colonies were mixed with transfected and non transfected cells. Further research will be needed for selection and establishment of gene transfected cells.

Method Comparison of Chemical-Resistance Level Determination and Field Resistance of Two-Spotted Spider Mite, Tetranychus urticae Koch to Benzomate, Cyhexatin, and Dicofol (점박이응애의 약제저항성(藥劑抵抗性) 수준결정방법(水準決定方法) 비교(比較)와 Benzomate, Cyhexatin 및 Dicofol 저항성(抵抗性) 조사(調査) 연구(硏究))

  • Lee, S.C.;Kim, W.Y.;Kim, S.S.
    • Korean journal of applied entomology
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    • v.25 no.3 s.68
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    • pp.133-138
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    • 1986
  • This study was conducted to compare the methods of slide dip and leaf dip techniques in the resistant level determination of spider mites to acaricides used; .and to investigate resistant levels of two-spotted spider mite, Tetranychus urticae Koch to benzomate, dicofol and cyhexatin in Honam region of Korea. Comparisons of the resistant strains and susceptible strain based on $LC_{50}$ values were summarized. Using the slide dip and leaf dip techniques with benzomate acaricide, the Keumchun strain of T. urticae showed 75-and 69-fold resistant levels, respectively, as compared with the Kwangju susceptible strain, but the Hackyo strain was 38-and 35-fold resistant levels, respectively. There was no significant difference (P>0.05) between the slide dip and leaf dip methods. With dicofol acaricide, resistant levels of the Keumchun strain of T. urticae were 71 times by using the slide dip and 74 times by using the leaf dip as resistant as the Kwangju susceptible strain while the Hackyo strain was 24 times by the former method and 23 times by the later method. There was also no significant difference (P>0.05) between both the methods. The slide dip technique is more recommendable than the leaf dip technique for determination of degree of chemical resistance to the spider mites with accuracy and less variation of the results. Using the slide dip method with cyhexatin, the Bia, Keumchun and Hackyo strains of T. urticae were 19-, 18- and 9-fold resistant levels, respectively, over the Kwangju susceptible strain.

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Comparison of Plant Community Structures in Cut and Uncut Areas at Burned Area of Mt. Gumo-san (금오산(金烏山)의 산화지(山火地)에서 벌목지(伐木地)와 비벌목지(非伐木地)의 식물(植物) 군집구조(群集構造) 비교(比較))

  • Che, Sang-Hoon;Kim, Woen
    • Journal of Korean Society of Forest Science
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    • v.86 no.4
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    • pp.509-520
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    • 1997
  • This is a report on the early vegetation, plant community structure, and secondary succession of cut and uncut sites of burned areas in Mt. Gumo-sun. The forest fire occurred on April, 1994 and the pine forest and its floor vegetation were burned down. The investigation was carried out from April, 1995 to October, 1996. The results are summarized as follows : The floristic composition of cut and uncut sites of burned area and unburned area were composed of 32, 36, and 34 kinds of vascular plants respectively. The biological spectra showed the $H(G)-D_1-R_5-e$ type, $H(M)-D_1-R_5-e$ and $M(N)-D_1-R_5-e$ in cut, uncut, and unburned site respectively. The dominant species based on $SDR_3$ of the cut site were Miscanthus sinensis var. purpurascens(100.00). Caret humilis(52.27), Quercus serrata(51.19) and Lysimachia clethroides(39.40), however, in the uncut sites the dominant species were Quercus acutissima(56.91), Pinus densiflora(26.83) in the tree layer, Quercus serrata(50.43), Lindera glauca(40.51), Lespedeza bicolor(37.85) in the shrub layer, and Miscanthus sinensis var. purpurascens(72.27), Pteridium aquilium var. latiusculum(60.92), Carex humilis(63.63) in the herb layer. Pinus densiflora(99.88), Miscanthus sinensis var. purpurascens(82.74), Quercus serrata(77.47) and Carex humilis(74.02) were dominant in the unburned site. The species diversity(H) and evenness index(e) were 1.05, 0.70 and 1.32, 0.85 in the cut and uncut site, respectively and 0.22, 0.63 in the unburned site. Dominance index(C) was 0.15, 0.06 and 0.96 in the cut, uncut site and unburned site, respectively. Degree of succession(DS) was 345.19, 747.47 and 674.34 in cut, uncut and unburned site, respectively. The index of similarity(CCs) was 0.66 between cut and uncut sites, 0.50 between unburned and cut sites and 0.61 between unburned and uncut sites. The amount of exchangeable sodium, calcium, magnesium and soil pH were increased, but the amount of organic matter, available phosphous, total nitrogen, total carbon and exchangeable potassium were decreased in cut site after fire.

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STUDY ON THE NUISANCE IN THE ANIMAL FARM (축산공해(畜産公害)에 관(關)한 연구(硏究) -양돈업(養豚業)을 중심(中心)으로)

  • Jeon, C.G.;Kim, J.W.;Ra, G.Y.;Kim, K.J.
    • Korean Journal of Agricultural Science
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    • v.2 no.1
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    • pp.233-240
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    • 1975
  • This excrement was conducted to investigate the nuisance of swine excrement. And excrements were collected from 27 heads of swine Euseong swine breeding center and Hongdo-dong swine farm and amounts of faces and urine, nitrogen, phosphorus, and kalium contents and number of parasite were investigated, and for the purpose to clean of excrementing matter, ferment substance and moist soil added to swine excreting matter and floating matter. Heat decresement amount and biological oxygen demand were investigated. And the result obtained were as follows. 1) Average amount of daily excreting faces per swine was 500-2700g from Euseong swine breeding center and 450-2500g from Hongdo-dong swine farm and a mount of excreting urine was 450-4500g from both of farms. The rate of excrements between faeces and urine was 1:1 2) Nitrogen contents were 0.802 ppm in urine and 0.514 ppm in faces, phosphorus were 2.261 % in urine and 0.073% in faeces and kalium contents were 1.094% in and 6.0467% in faeces. 3) Strongyloides ransomi and intestinal modular worm eggs showed the highest of eggs number was the result of observation in parasites, and ascaris showed the next it, whip worm and lung worm showed the lowest. 4) The effect ferment substance additive, amount of dry matter, floating matter, heat decreasement amount and biological oxygen demand was decreased but not significant was appeared. And the effect of moist soil additive, it was more effective that fermented substance additive for the fermentation of swine excrement.

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The Effect of Ginseng Saponins on the Biosynthesis of Prostaglandins (인삼 Saponin이 Prostaglandin 대사에 미치는 영향)

  • Park C.W.;Lee S.H.
    • Proceedings of the Ginseng society Conference
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    • 1988.08a
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    • pp.77-80
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    • 1988
  • The effects of ginseng saponins and some phenolic acids on the in vitro biosynthesis of prostaglandins was examined in order to identify the role of some ginseng components on the regulaion of arachidonic acid metabolism. The productions of prostaglandin $E_2(PGE_2).$ prostaglandin $F_2{\alpha}(PGF_2{\alpha}).$ thromboxane $B_2(TxB_2)$ and 6-keto-prostaglandin $F_1{\alpha}(6-keto-PGF_1{\alpha})$ from $[^3H]-arachidonic$ acid were evaluated with rabbit kidney microsome. human platelet homogenate and bovine aortic microsome. The amounts of the total cyclooxy-genase products from arachidonic acid did't show significant changes in the presence of ginseng saponins. Panaxadiol. panaxatriol and all of the ginsenosides used in these experiments reduced the formation of $TxB_2.$ while increased the $6-keto-PGF_1{\alpha}$ production dose dependently. Ginseng saponins did't inhibit the ADP($10{\mu}M$) induced platelet aggregation. but sodium arachidonate (0.5 mM) induced platelet aggregation. but sodium arachidonate (0.5 mM) induced platelet aggregation was signiticantly inhibited. These findings suggest that ginseng saponins seem to playa role in the regulation of the arachidonate metabolism. probably by affecting the divergent biosynthetic pathway of prostaglandins from endoperoxide.

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Development of in-situ Sintered Ni-Al Alloy Anode for Molten Carbonate Fuel Cell (용융탄산염 연료전지용 in-situ 소결된 Ni-Al 합금 연료극 개발)

  • Chun, H.A.;Yoon, S.P.;Han, J.;Nam, S.W.;Lim, T.H.
    • Journal of the Korean Electrochemical Society
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    • v.9 no.3
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    • pp.124-131
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    • 2006
  • For commercialization of molten carbonate fuel cell (MCFC), it has some problems to be overcome such as decrease of porosity and thickness of the anode under the operating condition (at $650^{\circ}C$ and working pressure of more than 2 $kg_f/cm^2$). Recently, Ni-Al alloy anode has been proposed to replace the conventional Ni-Cr anode as an alternative material to resist a creep and inhibit the sintering. The objective of this research is to sinter the green sheet of Ni-Al alloy anode during single cell pre-treatment process, which has several advantages like cost down and simplification of manufacturing process. However, the Ni-Al alloy anode prepared with a conventional pre-treatment process showed the phase separation of Ni-Al alloy and formation of micropore(${\leqq}0.4{\mu}m$), resulting in low creep resistance and high electrolyte re-distribution. In order to prevent the Ni-Al alloy anode from phase-separating, nitrogen gas was used in the process of pre-treatment. Introducing the nitrogen, the phase separation from Ni-Al alloy into nickel and alumina was minimized and increased creep resistance. However, there was some micropore formation on the surface of Ni-Al alloy anode during the cell operation due to creation of lithium aluminate. Addition of more amount of electrolyte into a cell, especially at cathode, made the cell performance stable for 2,000 hrs. Consequently, it was possible to make the Ni-Al alloy anode with good creep resistance by the modified in-situ sintering technique.

Determination of Energy Requirements for Maintenance in Hanwoo Steers (거세한우의 유지에너지 요구량 결정)

  • Kim, K.H.;Oh, Y.G.;Kim, W.;Lee, S.C.;Shin, K.J.;Jeon, B.T.
    • Journal of Animal Science and Technology
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    • v.46 no.2
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    • pp.193-200
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    • 2004
  • This experiment was carried out to determine energy requirements for maintenance of Hanwoo steers. Nine Hanwoo steers weighing 376.6$\pm$12.5kg were used in this experiment and fed rice straw(44%) and concentrate (56%) at three different energy levels; 0.8 times maintenance(0.8M), 1.2 times(1.2M) and 1.6 times(1.6M), respectively. Dry matter intake was 48.5, 65.9 and 86.5g/$BW^{0.75}$ for 0.8M, 1.2M and 1.6M, respectively. Increase in energy intake with the increased DM intake did not affect digestibilities of dry matter, organic matter, crude protein, crude fiber, crude fat and nitrogen-free extract. Gross energy intake averaged 190.8, 255.8 and 340.9kcal/BW0.75 for 0.8M, 1.2M and 1.6M, respectively. Energy loss was 41% feces and 0.6${\sim}$1.5% urine of gross energy intake. Further, energy loss from methane produced during rumen fermentation was 5${\sim}$9%, while body heat loss averaged 40${\sim}$60%. Intercept of the regression equation between ME intake and retained energy indicated that energy requirement for maintenance was 124.3kcal $ME/BW^{0.75}$.

Pharmacological Evaluation of the Mechanism of ${\alpha}-Adrenoceptor-Mediating$ Sleep in Chickens (${\alpha}$-아드레나린 수용체의 매개에 의한 병아리 수면에 대한 약리학적 고찰)

  • Jeong, S.H.;Sohn, U.D.;Song, C.S.;Hong, K.W.
    • The Korean Journal of Pharmacology
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    • v.20 no.2
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    • pp.15-21
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    • 1984
  • It was aimed to study the effects of ${\alpha}_2-adrenoceptor$ agonists on the sleeping time in $one{\sim}two-day-old$ chickens. Furthermore, it was also evaluated whether ${\alpha}_1-adrenoceptor$ agonist and antagonist might affect the sleeping in the chickens and discussed in relation with opiate receptor. 1) Guanabenz, clonidine, guanfacine and B-HT 933 decreased the latency of the loss of righting reflex in a dose-dependent manner, but B-HT 920 and oxymetazoline slightly prolonged it. 2) ${\alpha}_2-Adrenoceptor$ agonists produced dose·related increase in sleeping time. The potency was guanabenz>clonidine>oxymetazoline${\geq}$B-HT 933${\geq}$B-HT 920>guanfacine in this order. 3) ${\alpha}_2-Adrenoceptor$ antagonists decreased guanabenz-induced sleeping time in a dose ·dependent manner. The rank order of ${\alpha}_2-adrenoceptor$ antagonists was yohimbine>rauwolscine>piperoxan${\geq}$RX 781094. 4) Sleeping time caused by both ethanol and hexobarbital was not affected by yohimbine in chickens. 5) Methoxamine and phenylephrine showed little significant effect on the guanabenz-induced sleeping time. However, prazosin increased it. Paradoxically, corynanthine rather caused to decrease it. These results suggest that the stimulation of central ${\alpha}_2-adrenoceptor$ mediates sleeping, however it is remained uncertain in the role of central ${\alpha}_1-adrenoceptor$ in chickens. In addition, the one~two-day-old chickens may be considered as a useful, inexpensive and simple experimental model to evaluate the in vivo pharmacological action of the ${\alpha}_2-adrenoceptor$ agonist and antagonist related to sedation.

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