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Biochemical Properties of Lactate Dehydrogenase Eye-Specific C4 Isozyme: Lepomis macrochirus and Micropterus salmoides (젖산탈수소효소 eye-specific C4 동위효소의 생화학적 특성: 파랑볼우럭(Lepomis macrochirus)과 큰입우럭(Micropterus salmoides))

  • Yum, Jung-Joo;Ku, Bo-Ra
    • Journal of Life Science
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    • v.22 no.2
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    • pp.209-219
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    • 2012
  • The properties of lactate dehydrogenase (LDH, EC 1.1.1.27) eye-specific $C_4$ isozyme were studied by polyacrylamide gel electrophoresis, Western blotting, immunoprecipitation, and enzyme kinetics. Furthermore, we proposed the optimal conditions for measuring the activity of LDH eye-specific $C_4$ isozyme. The isozymes were detected in the cytosol of eye tissues from Lepomis macrochirus and Micropterus salmoides and were more similar to the $A_4$ than the $B_4$ isozyme. LDH/CS in the eye tissue of L. macrochirus was increased in September, so the ratio of anaerobic metabolism was high. The electrophoretic patterns of mitochondrial LDH were similar to those of cytosolic LDH in the eye tissues of L. macrochirus and Micropterus salmoides. LDH eye-specific $C_4$ isozyme from eye tissue was purified by preparative native-PAGE. The activities of LDH eye-specific $C_4$ isozymes in L. macrochirus and M. salmoides were reduced at concentrations greater than 0.2 mM and 0.1 mM of pyruvate, respectively. These concentrations remained at 5.2% and 15.8% as a result of the inhibition by 10 mM of pyruvate, so the degree of inhibition was very high. The LDH activities of eye tissues were reduced at concentrations greater than 22 mM and 24 mM of lactate, respectively, in L. macrochirus and M. salmoides. The ${K_m}^{PYR}$ of eye-specific $C_4$ was 0.088 mM in L. macrochirus and it was 0.033 mM in M. salmoides. The activities of cytosolic and mitochondrial eye-specific $C_4$ isozymes were high in ${\alpha}$-ketobutyric acid. Furthermore, the activities of eye tissue and eye-specific $C_4$ isozyme had to be measured with 0.5 mM of pyruvate and a buffer solution of pH 7.5. As a conclusion, the eye-specific $C_4$ isozyme in M. salmoides has a high affinity for pyruvate and exhibits maximum activity at a lower concentration of pyruvate and at higher concentration of lactate than that in L. macrochirus. Therefore, it seems that the energy produced by the LDH eye-specific $C_4$ isozyme in M. salmoides was used at the first stage of predatory behavior.

Effects of Electric Stimulation and Activation Conditions on the Fusion and Development of Porcine Somatic Cell Nuclear Transfer Embryos (전기적 융합과 활성화 방법이 돼지 체세포 복제수정란의 체외발달에 미치는 영향)

  • 정기화
    • Journal of Embryo Transfer
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    • v.19 no.1
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    • pp.43-51
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    • 2004
  • The present study was conducted to investigate the effects of fusion and/or activation protocol on in vitro development of porcine somatic cell nuclear transfer (SCNT) embryos. Porcine fetal fibroblast cells were transferred into the perivitelline space of enucleated in vitro matured oocytes. Cell fusion and activation were induced simultaneous fusion/activation (SA) or delayed activation (DA) with or without cytochalasin B (CB) treatment with electic pulses in 0.28 M mannitol-based medium. The SCNT embryos were cultured in vitro for 7 days and stained with Hoechst 33342 to determine the number of nuclei. After 7 days culture, cleavage and blastocyst formation rates were 72.4% and 7.6% in SCNT and 76.3% and 20.4% in parthenotes. To examine the effect of electric field strengths on development of SCNT embryos, oocytes were fused two pulses of 110 V/mm, 130 V/mm or 150 V/mm for 30 sec post-injection. The fusion and cleavage rates in 130 V/mm group (70.2% and 72.6%) and 150 V/mm group (72.6% and 70.5%) were higher (P<0.05) than 110 V/mm group (47.1% and 48.6%), respectively. However, the rate of embryos developing to the blastocyst stage (8.1%, 9.7% and 10.7%) were not different among three groups. The cleavage rates and the blastcyst formation rates were not different among three treatment groups (SA group, 71.4% and 9.7%; SA+CB treatment group, 74.7% and 8.0%; DA+CB treatment group, 70.8% and 11.2%, respectively). And, no different in the number of cells in blastocysts was observed among the three groups (22.5$\pm$12.8, 23.3$\pm$11.2 and 21.6$\pm$10.4, respectively). These result suggest that two pulses of 130 V/mm or 150 V/mm for 30 sec with SA treatment or DA treatment are enough for fusion/activation of porcine somatic cell nuclear transfer (SCNT) embryos to develop to the blastocyst stage.

Characteristics of Manure and Estimation of Nutrient and Pollutant of Holstein Dairy Cattle (홀스타인 젖소 분뇨의 특성과 비료성분 및 오염물질 부하량 추정)

  • Choi, D.Y.;Choi, H.L.;Kwag, J.H.;Kim, J.H.;Choi, H.C.;Kwon, D.J.;Kang, H.S.;Yang, C.B.;Ahn, H.K.
    • Journal of Animal Science and Technology
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    • v.49 no.1
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    • pp.137-146
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    • 2007
  • This study was conducted to determine fertilizer nutrient and pollutant production of Holstein dairy cattle by estimating manure characteristics. The moisture content of feces was 83.9% and 95.1% for urine. The pH of feces and urine were in the ranges of 7.0~7.4 and 7.5~7.8, respectively. The average BOD5, COD, SS, T-N, T-P concentrations of the dairy feces were 18,294, 52,765, 102,889, 2,575, and 457mg/ℓ, respectively. Dairy urine showed lower levels of BOD5(5,455mg/ℓ), COD(8,089mg/ℓ), SS(593mg/ℓ), T-N(3,401mg/l), and T-P(13mg/ℓ) than feces. The total daily produced pollutant amounts of a dairy cow were 924.1g(Milking cow), 538.8g(Dry cow), 284.4g(Heifer) of BOD5, 2,336.5g (Milking cow), 1,651.8g(Dry cow), 734.1g(Heifer) of COD and 4,210.1g(Milking cow), 2,417.1g(Dry cow), 1,629.1g(Heifer) of SS and 194.8g(Milking cow), 96.4g(Dry cow), 58.3g(Heifer) of T-N and 24.0g(Milking cow), 10.2g(Dry cow), 6.1g(Heifer) of T-P. The calculated amount of pollutants produced by a 450kg dairy cow for one year were 181.3kg of BOD5, 492.5kg of COD, 899.9kg of SS, 36.0kg of T-N and 4.1kg of T-P. The total yearly estimated pollutant production from all head(497,261) of dairy cattle in Korea is 90,149 tons of BOD5, 244,890 tons of COD, 447,491 tons of SS, 17,898 tons of T-N and 2,008 tons of T-P. The fertilizer nutrient concentrations of dairy feces was 0.26% N, 0.1% P2O5 and 0.14% K2O. Urine was found to contain 0.34% N, 0.003% of P2O5 and 0.31% K2O. The total daily fertilizer nutrients produced by dairy cattle were 197.4g (Milking cow), 97.4g(Dry cow), and 57.9g(Heifer) of Nitrogen, 54.2g(Milking cow), 22.2g(Dry cow), and 14.2g(Heifer) of P2O5 and 110.8g(Milking cow), 80.4g (Dry cow), and 39.5g(Heifer) of K2O. The total yearly estimated fertilizer nutrient produced by a 450kg dairy animal is 36.2kg of N, 8.8kg of P2O5, 24.6kg of K2O. The estimated yearly fertilizer nutrient production from all dairy cattle in Korea is 18,000 tons of N, 4,397 tons of P2O5, 12,206 tons of K2O. Dairy manure contains useful trace minerals for crops, such as CaO and MgO, which are contained in similar levels to commercial compost being sold in the domestic market. Concentrations of harmful trace minerals, such as As, Cd, Hg, Pb, Cr, Cu, Ni, Zn, met the Korea compost standard regulations, with some of these minerals being in undetected amounts.

Soil Microbial Flora and Chemical Properties as Influenced by the Application of Pig Manure Compost (돈분퇴비의 시용이 배추재배지 토양의 미생물상 및 화학성에 미치는 영향)

  • Weon, Hang-Yeon;Kwon, Jang-Sik;Suh, Jang-Sun;Choi, Woo-Young
    • Korean Journal of Soil Science and Fertilizer
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    • v.32 no.1
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    • pp.76-83
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    • 1999
  • Studies were conducted during 2 months from May of 1997 to evaluate the effects of pig manure compost(PMC) on soil microbial flora. To do so, a field experiment of Chinese cabbage(Brassica campestris L.) was conducted in a randomized block design on a sandy loam soil and microbial floral characteristics in soils were analyzed. Treatments to control included the application of PMC at (A) $8Mg\;ha^{-1}$CM-8), (B) $29Mg\;ha^{-1}$(CM-2,9), and (C) $57Mg\;ha^{-1}$(CM-57), and of chemical fertilizer(D) at $320N-80P_2O_5-200K_2O\;kg\;ha^{-1}$(NPK). In each treatment, the rhizosphere and non-rhizosphere soils were tested for the analysis of microbial populations. The populations of bacteria, actinomycetes, and fungi increased in soils with the applications of PMC and chemical fertilizer, but that of Bacillus sp. decreased. However, the population of fluorescent Pseudomonas sp. was reduced in NPK plots only. With increasing application rates of PMC, the number of colony forming units(cfu) of bacteria (Pseudomonas sp. and actinomycetes) and fungi increased. in all PMC-treated plots, the population density peaked at early growth stage for bacteria(including Bacillus sp.), at late growth for fluorscent Pseudomonas sp., and at harvest for fungi and actinomycetes. The rhizosphere effect was greatest for fluorscent Pseudomonas sp. As the application rates of PMC increased, Total N, organic matter, available phosphate, and exchangeable -K, -Ca, and -Mg increased compared to control, but soil pH was lowered. In NPK plots, EC was 3.4-fold and exchangeable K was 5-fold higher than control.

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Biological activity of shizukanols isolated from Chloranthus japonicus roots (홀아비꽃대(Chloranthus japonicus) 뿌리로부터 분리한 shizukanol들의 생물활성)

  • Park, Mi-Ran;Kim, Hae-Young;Choi, Gyung-Ja;Lee, Seon-Woo;Jang, Kyoung-Soo;Kim, Jin-Seog;Hong, Kyung-Sik;Park, No-Joong;Cho, Kwang-Yun;Kim, Jin-Cheol
    • The Korean Journal of Pesticide Science
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    • v.8 no.4
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    • pp.338-346
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    • 2004
  • The methanol extract of Chloranthus japonicus roots effectively controlled the development of rice blast(Magnaporthe grisea), rice sheath blight(Corticium sasaki), tomato pay mold(Btrytis cinerea), tomato late blight(Phytophthora infestans), and wheat leaf rust(Puccinia recondita). From the methanol extract of C. japonicus roots, three antifungal substances were isolated. Their chemical structures were determined to be shizukanols B, C, and D mainly by mass and NMR spectral data. Among the three substances, shizukanol C showed the strongest inhibitory activity against mycelial growth of the plant pathogenic fungi tested; it completely inhibited mycelial growth of M. grisea. Colletotrichum gloeosporioides, and C. acutatum at concentrations of more than $12.5{\mu}g$/ and P. infestans at concentrations of more than $3.13{\mu}g/m\ell$. They also controlled effectively the development of rice blast and wheat leaf rust. On the other hand, they caused phytotoxic symptoms on barley leaves and inhibited the growth of duckweed (Lemna paucicostata) with $EC_{50}$ values of $30.0{\mu}g/m\ell$ for shizukanol B, $49.9{\mu}g/m\ell$ for shizukanol C, and $154{\mu}g/m\ell$ for shizukanol D. In addition, shizukanol C showed an insecticidal activity against brown planthopper (Nilaparavata lugens), peen peach aphid (Myzus persicae), diamond-back moth (Plutella xylostella), and tobacco cutworm (Spodoptera litura) of the 5 arthropod pests tested with mortality values of more than 60% at a concentration of $1,000{\mu}g/m\ell$.

The Study of Solid Waste Compost Development for Reclaiming Damage Soil in Forest (산림훼손토양 복원을 위한 부숙토 개발 연구)

  • Na, Seung-Ju;Chang, Ki-Woon;Yang, Hui-Young;Jeon, Han-Ki;Lee, Jong-Jin
    • Journal of the Korea Organic Resources Recycling Association
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    • v.13 no.2
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    • pp.107-120
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    • 2005
  • To study the development of solid waste compost to use sewage sludge and paper mill sludge for reclaiming damage soil in forest, the changes of temperature, moisture, chemical properties, heavy metals and harmful compound during the aerobic decomposition were investigated, and the compost decomposition of final products investigated the round paper chromatography method and G.I(Germination index) value. The results were summarized as follows. Temperature was changed a little during early 5days because of air temperature too low. That was rapidly increased to over $50^{\circ}C$ at 4days after first turning and then decreased gradually fallen to $40{\sim}50^{\circ}C$ at 15days after aerobic decomposition in A and C treatments. The second turning was conducted at 18 days after aerobic decomposition, and then the temperature was little changed. At the compare first with terminal product, The moisture content was decreased all treatments but the change was little in A and B treatments. pH was decreased to below 1 in all treatments. EC was increased to below 5dS/m. The content of total carbon, C/N ratio, $NH_4{^+}-N$ were decreased with 4~7%, below 8 and below 500mg/kg in all treatments, respectively. The content of total nitrogen, $NO_3{^-}-N$, CEC were increased with below 0.5%, below 173mg/kg and over $30cmol^+/kg$ in all treatments, respectively. The content of heavy metals and harmful compound were similar during aerobic decomposition and suited to standard of 가 grade in all treatments. The result of round paper chromatography method and G.I. value, The C treatment concluded well aerobic decomposition. Especially, the G.I. value in C treatment was 64.1 and 66.2 at cabbage and grass, respectively.

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Composition of Crushed Oyster Shell and its Application Effect on Vegetables (굴껍질분(紛)의 화학성(化學性) 및 작물(作物)에 대한 시용효과(施用效果))

  • Kim, Jong-Gyun;Lee, Han-Saeng;Cho, Jea-Gyu;Lee, Young-Han
    • Korean Journal of Soil Science and Fertilizer
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    • v.28 no.4
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    • pp.350-355
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    • 1995
  • This study was conducted to use oyster shells, which have caused environmental problems in the coastal of Korea, as an agricultural material after processing. Physico-chemical components and neutralizing amount on the Ihyun silt loam of crushed oyster shell and slaked lime were examined. In field experiment, the properties of the soil, growth and yield of lettuce, cabbage, spinach, onion, red pepper and soybean were examined by the treatments of the shell(3.68ton/ha) or the lime (2.76ton/ha) with a randomized block design. Particle size of crushed oyster shell consisted of 73.4% of 1~60mesh and 26.6% larger then 61 mesh and contents of CaO, OM, and $P_2O_5$, etc. were 55.5%, 1.3%, and 0.29%, respectively. The requirement of the shell to neutralize the soil was 130~135% of the lime, but after 24months, it was the same. The application of the shell increased the contents of available $P_2O_5$ and $SiO_2$ exchangeable Ca in used soil. The shell tratment increased the leaf height, leaf width, etc. of the examined plants, and the yields 6~154% according to examined plants, as compared with the nonliminged, indicating that the shell possesses a great potential as an agricultural material with the same effectiveness as the lime.

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