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Deterimination of an Optimal Time Point for Analyzing Transcriptional Activity and Analysis of Transcripts of Avian Influenza Virus H9N2 in Cultured Cell (배양세포에서 Semi-quantitative RT-PCR에 의한 조류인플루엔자 H9N2의 전사활성 분석 최적 시기 결정 및 전사체 분석)

  • Na, Gi-Youn;Lee, Young-Min;Byun, Sung-June;Jeon, Ik-Soo;Park, Jong-Hyeon;Cho, In-Soo;Joo, Yi-Seok;Lee, Yun-Jung;Kwon, Jun-Hun;Koo, Yong-Bum
    • Korean Journal of Microbiology
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    • v.45 no.3
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    • pp.286-290
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    • 2009
  • The transcription of mRNA of avian influenza virus is regulated temporally during infection. Therefore, the measurement of transcript level in host cells should be performed before viral release from host cells because errors can occur in the analysis of the transcript levels if the viruses released from the infected cells re-infect cells. In this study, the timing of viral release was determined by measuring the level of viral RNA from viruses released from H9N2-infected chicken fibroblast cell line UMNSAH/DF-1 by semi-quantitative RT-PCR. The viral genomic RNA was isolated together with mouse total RNA which was added to the collected medium as carrier to monitor the viral RNA recovery and to use its GAPDH as an internal control for normalizing reverse transcription reaction as well as PCR reaction. It was found that viral release of H9N2 in the chicken fibroblast cell line UMNSAH/DF-1 took between 16 and 20 h after infection. We measured all 8 viral mRNA levels. Of the 8 transcripts, 7 species of viral mRNAs (each encoding HA, NA, PB1, PB2, NP, M, NS, respectively) except PA mRNA showed robust amplification, indicating these mRNA can be used as targets for amplification to measure transcript levels. These results altogether suggest that the method in this study can be used for screening antiviral materials against viral RNA polymerase as a therapeutic target.

Identification of a new marine bacterium Ruegeria sp. 50C-3 isolated from seawater of Uljin in Korea and production of thermostable enzymes (대한민국 울진 연안 해양에서 분리한 해양 미생물 Ruegeria sp. 50C-3의 동정 및 내열성 효소 생산)

  • Chi, Won-Jae;Kim, Jong-Hee;Park, Jae-Seon;Hong, Soon-Kwang
    • Korean Journal of Microbiology
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    • v.52 no.3
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    • pp.344-351
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    • 2016
  • A marine bacterium, designated as strain 50C-3, was isolated from a seawater sample collected from the East Sea of South Korea. The strain is a Gram-negative, aerobic, yellow colored polar-flagellated bacterium that grows at $20-50^{\circ}C$ and pH 5.5-8.5. Optimal growth occurred at $40-50^{\circ}C$, at pH 6.5-7.5, and in the presence of 2% (w/v) NaCl. Based on 16S rRNA gene sequence similarity, the isolate was considered to represent a member of the genus Ruegeria. The result of this analysis showed that strain 50C-3 shared 99.4% and 96.98% sequence similarity with Ruegeria intermedia CC-GIMAT-$2^T$ and Ruegeria lacuscaerulensis ITI-$1157^T$, respectively. Furthermore, strain 50C-3 showed clear differences from related strains in terms of several characteristics such as motility, carbon utilization, enzyme production, etc. The DNA G+C content was 66.7 mol%. Chemotaxonomic analysis indicated ubiquinone-10 (Q-10) as the predominant respiratory quinone. Based on phenotypic, chemotaxonomic, and phylogenetic characteristics, the isolate represents a novel variant of the Ruegeria intermedia CC-GIMAT-$2^T$, for which we named Ruegeria sp. 50C-3 (KCTC23890=DSM25519). Strain 50C-3 did not produce cellulase and agarase, but produced alkaline phosphatase, ${\alpha}$-galactosidase, and ${\beta}$-galactosidase. The three enzymes showed stable activities even at $50^{\circ}C$ and thus regarded as thermostable enzymes. Especially, the ${\beta}$-galactosidase activity enhanced by 1.9 times at $50^{\circ}C$ than that at $37^{\circ}C$, which may be very useful for industrial application.

Protoplast Formation, Regeneration and Reversion in Pleurotus ostreatus and P. sajor-caju (느타리버섯과 여름느타리버섯의 원형질체(原形質體) 나출(裸出)과 재생(再生))

  • Go, Seung-Joo;Shin, Gwan-Chull;Yoo, Young-Bok
    • The Korean Journal of Mycology
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    • v.13 no.3
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    • pp.169-177
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    • 1985
  • The studies were carried out to obtain the basic data for maximizing the protoplast yields from the mycelia of P. ostreatus and P. sajor-caju. Some factors affecting the regeneration of the protoplast of both species and the productivity of their reversion were also examined. The maximum yields of protoplasts were obtained from four days cultured mycelia of both species on cellophan membrane placed on the surface of PSA or MCM media in a petri dish. The optimal concentration of lytic enzyme Novozym 234 for protoplast releasing was 5 mg per ml of 0.5 M phosphate buffer solution with 0.6 M sucrose or 0.6 M $MgSO_4$ at pH 6.0. The greatest number of protoplasts was released 3 hours after incubation of the mycelia of P. ostreatus and after 4 hours for the P. sajor-caju in the lytic enzyme solution. Among the osmotic stabilizer solutions tested 0.6 M sucrose and 0.6 M KCl showed the best regeneration rates of the protoplasts of both species. When 0.75 % agar solution was over-layed on the regeneration media immediately after inoculation of the protoplast the regeneration rates were greatly enhanced. The ampicillin added to the agar solution prevented bacteria from infection. The reverted isolates produced the sporophores and basidial spores just like their parents without any mutations when they were cultivated in a broad mouth bottle with sawdust substrates.

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Inhibitory effect of Hypericum ascyron on pro-inflammatory responses in lipopolysaccharide-induced Raw 264.7 Cells (Lipopolysaccharide로 유도된 Raw 264.7 cell에서 물레나물(Hypericum asctron)의 Pro-inflammatory 억제 효과)

  • Hong, Eun-Jin;Park, Hye-Jin;Kim, Na-Hyun;Jo, Jae-Bum;Lee, Jae-Eun;Lim, Su-Bin;Ahn, Dong-Hyun;Jung, Hee-Young;Cho, Young-Je
    • Journal of Applied Biological Chemistry
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    • v.60 no.4
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    • pp.363-372
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    • 2017
  • Hypericum ascyron has long been used as medicinal plant and recent studies reported that H. ascyron has anti-diabetic, anti-oxidant, and anti-bacterial effects. In this study, inhibitory effect from H. ascyron on pro-inflammatory responses has been investigated. H. ascyron was extracted at optimal extraction condition. Total phenolic contents in water and 90% ethanol were 29.75 and 31.82 mg/g, respectively. Hyaluronidase inhibitory activity of H. ascyron extracts ($50-200{\mu}g/mL$ phenolics) was 0.00-14.81% and 15.33-47.49%, respectively. In cell viability, cell toxicity was shown at concentration of $100{\mu}g/mL$ and $30{\mu}g/mL$ of water and 90% ethanol extract. Therefore, $10-50{\mu}g/mL$ in water extracts and $5-20{\mu}g/mL$ in ethanol extracts was selected each for further study. Inducible nitric oxide synthase (iNOS) derived nitric oxide (NO) and cyclooxygenase (COX)-2-derived prostaglandin $E_2$ ($PGE_2$) protein expression inhibitory effect of extracts were inhibited in a dose dependent manner, significantly. Also, the pro-inflammatory cytokines inhibitory effect such as tumor necrosis $factor-{\alpha}$, nterleukin (IL)-6 and $IL-1{\beta}$ were decreased in the dose dependent manner. The results indicate that H. ascyron extracts reduced inflammatory responses in lipopolysaccharide-induced 264.7 cells via the regulation of the iNOS, COX-2, NO, $PGE_2$, and pro-inflammatory cytokines. Therefore, H. ascyron extracts have significant anti-inflammatory effect and a source as therapeutic materials.

Prospective for Successful IT in Agriculture (일본 농업분야 정보기술활용 성공사례와 전망)

  • Seishi Ninomiya;Byong-Lyol Lee
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.6 no.2
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    • pp.107-117
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    • 2004
  • If doubtlessly contributes much to agriculture and rural development. The roles can be summarized as; 1. to activate rural areas and to provide more comfortable and safe rural life with equivalent services to those in urban areas, facilitating distance education, tole-medicine, remote public services, remote entertainment etc. 2. To initiate new agricultural and rural business such as e-commerce, real estate business for satellite officies, rural tourism and virtual corporation of small-scale farms. 3. To support policy-making and evaluation on optimal farm production, disaster management, effective agro-environmental resource management etc., providing tools such as GIS. 4. To improve farm management and farming technologies by efficient farm management, risk management, effective information or knowledge transfer etc., realizing competitive and sustainable farming with safe products. 5. To provide systems and tools to secure food traceability and reliability that has been an emerging issue concerning farm products since serious contamination such as BSE and chicken flu was detected. 6. To take an important and key role for industrialization of farming or lam business enterprise, combining the above roles.

Effect of High Temperature, Daylength, and Reduced Solar Radiation on Potato Growth and Yield (고온, 일장 및 저일사 조건이 감자 생육 및 수량에 미치는 영향)

  • Kim, Yean-Uk;Lee, Byun-Woo
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.18 no.2
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    • pp.74-87
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    • 2016
  • Potato phenology, growth, and yield are projected to be highly affected by global warming in the future. The objective of this study was to examine the responses of potato growth and yield to environmental elements like temperature, solar radiation, and daylength. Planting date experiments under open field condition were conducted using three cultivars differing in maturity group (Irish Cobbler and Superior as early; Atlantic as mid-late maturing) at eight different planting dates. In addition, elevated temperature experiment was conducted in four plastic houses controlled to target temperatures of ambient temperature (AT), $AT+1.5^{\circ}C$, $AT+3^{\circ}C$, and $AT+5^{\circ}C$ using cv. Superior. Tuber initiation onset was found to be hastened curve-linearly with increasing temperature, showing optimum temperature around $22-24^{\circ}C$, while delayed by longer photoperiod and lower solar radiation in Superior and Atlantic. In the planting date experiments where the average temperature is near optimal and solar radiation, rainfall, pest, and disease are not limiting factor for tuber yield, the most important determinant was growth duration, which is limited by the beginning of rainy season in summer and frost in the late fall. Yield tended to increase along with delayed tuber initiation. Within the optimum temperature range ($17^{\circ}-22^{\circ}C$), larger diurnal range of temperature increased the tuber yield. In an elevated temperature treatment of $AT+5.0^{\circ}C$, plants failed to form tubers as affected by high temperature, low irradiance, and long daylength. Tuber number at early growth stage was reduced by higher temperature, resulting in the decrease of assimilates allocated to tuber and the reduction of average tuber weight. Stem growth was enhanced by elevated temperature at the expense of tuber growth. Consequently, tuber yield decreased with elevated temperature above ambient and drop to almost nil at $AT+5.0^{\circ}C$.

Oocyte-sperm Binding Assay (OSBA) Technique for Rapid Q/C of IVF Culture Condition (체외수정용 배양조건의 신속한 Q/C를 위한 정자-난자 결합분석법(OSBA) 개발)

  • 정구민;신영수
    • Korean Journal of Animal Reproduction
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    • v.25 no.2
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    • pp.163-169
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    • 2001
  • OSBA(oocytes-sperm binding assay) is a tool developed for rapid test of optimal condition of IVF medium and protein source by binding ability of mouse sperm and egg. Mouse oocyte-cumulus complexes were prepared by removing of the cumulus cells with 0.1% hyaluronidase. 10$\pm$2 oocytes per 30 ${mu}ell$ medium drop were inseminated with 3 ${mu}ell$ sperm suspension and were cultured f3r 3 hours and 24 hours, respectively. And the oocytes were recovered gently and the No. of sperm bound on oocytes were counted. In the Exp. 1, the ratio of oocytes bound with one sperm at least were 60.2%(50/83), 2%(2/77) and 100%(79/79) in the medium with no protein, FBS(15%, v/v) and BSA(0.4%. w/v), respectively, Fetal bovine serum(FBS) seriously inhibited sperm binding on oocyte, although bovine serum albumin(BSA) promoted the binding ability. The inhibiting effect of FBS was dependent on the concentration of FBS. The sperm binding ability according to oocyte maturity was tested in the Exp. 2. There was no significant difference between Met. II (mature) and Met. I (intermediate mature) oocytes in the number of oocytes bound with sperm and the number of sperm bound on oocytes. Finally, in Exp. 3, two batches of Ham's F10 medium with good and poor quality by OSBA were tested (The ratios of embryos developed from PN 1-cell stage to hatched blastocyst; 25% vs. 70%). In the medium with good quality, sperm binding ability was significantly increased (P < 0.05). The ratio of oocytes bound with one sperm at least was 66% and 90% in the medium with poor and good quality, respectively. Conclusively, It was possible to test IVF medium condition rapidly and easily by OSBA.

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The Optimal TDN Levels of Concentrates and Slaughter Age in Hanwoo Steers (거세한우에 있어서 배합사료의 적정 TDN 수준과 도축 월령)

  • Kim, K.H.;Lee, J.H.;Oh, Y.G.;Kang, S.W.;Lee, S.C.;Park, W.Y.;Ko, Y.D.
    • Journal of Animal Science and Technology
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    • v.47 no.5
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    • pp.731-744
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    • 2005
  • Ninety Hanwoo steers(initial BW=167.2$\pm$13.4kg) were used to determine the effect of energy concentrations in concentrates and slaughter age on performance, carcass characteristics. Steers were allotted by BW to one of eighteen pens. Eighteen pens were randomly allotted to a low(70-70-71-72% for the growing, the early, the middle or the late fattening periods), medium(70-71-72-73%) or high(70-72- 73-74%) TDN level of concentrates. Five steers for each treatment of energy level were slaughtered every one month from 26 month of age to 31 month of age. Concentrates was fed restrictedly to achieve a predicted gain of 0.7-0.9kg from growing stage to middle fattening stage. All steers were fed orchard grass(Dactylis glomerata L.) hay as roughage during the growing period, fed rice straw gradually substituted for orchard grass hay during the early fattening period, and fed rice straw only thereafter. Overall body weight and feed intake were not affected by TDN levels of concentrates. Average daily gain for all treatments was higher than 0.9kg/d during the 19-21 month of age and decrease thereafter, but sustained above 0.7kg/d. Mean concentrates intake for all treatments was 1.0-1.3% of live BW during the growing period and 1.5% during the early fattening. Thereafter, it decreased up to 1.4% during the middle fattening and 1.0% during late fattening period. Delay of slaughter end point resulted in a gradual increase of rib-eye area, back fat thickness and marbling score, especially after slaughter age of 29 month there was significant increases(P<0.05). The appearance rate of 1+ and 1 grade related to the slaughter ages was 100% at 29, 30 and 31 months of age, whereas those at 26, 27 and 28 months were 93, 86 and 80%, respectively. Dressing rate was significantly(P<0.05) increased and rate of retailed cut weight significantly(P<0.05) decreased when slaughter age increased. In economic analysis, there was pronounced increase in net income up to 32-46% after slaughter age of 29 months. Under the conditions of this study, high TDN intake is not necessarily required for high quality Hanwoo meat production and slaughter age of 29 month might be the optimum for Hanwoo steers.

Effects of Dietary Crude Protein Level on Growth and Meat Quality of Korean Native Goats (사료의 조단백질 수준이 육성기 흑염소의 발육과 육질에 미치는 영향)

  • Choi, S.H.;Kim, S.W.;Park, B.Y.;Sang, B.D.;Kim, Y.K.;Myung, J.H.;Hur, S.N.
    • Journal of Animal Science and Technology
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    • v.47 no.5
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    • pp.783-788
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    • 2005
  • This study was conducted to investigate the optimal CP level in growing goat kids. Forty male goats were divided into four treatment groups fed diets containing CP 12, 14, 16, and 18% of concentrate feed, and rice straw, respectively. Results are summarized as follows. Average daily gain’s(ADG) of groups fed diets with CP 14% and CP 18% were 84.0 and 83.0g/d each, which were higher than group fed diets with CP 12% grown at 69.2g of ADG(P<0.05). Daily feed intakes of concentrate and rice straw were 590g and 45g each and there was no difference found between treatments. The amounts of feed required for body weight gain(g) were similar in the range of 7.0-7.3 for groups fed diets with CP 14%, 16% or 18%. But the group fed diets with CP 12% required somewhat more feed for gain at 8.8g. Dressing percentage of groups fed diet with CP 12% was 61.7%, which was higher than groups fed diets with CP 14% or CP 18%(P<0.05) but similar to that fed diet with CP 16%. Meat percentage from goats fed diet with CP 16% was 51.7%, which were higher than goats fed diets with CP 12% or CP 14%(P<0.05). Fat deposition of CP 12% group was higher than the other groups(P<0.05). Percentage of bone weight averaged 17.0% without any difference among treatment groups. Shear force and cooking loss, which are physical properties of goat meats, were lower in CP 16% and CP 14% groups than the other two groups. Water holding capacity of goats meats from CP 16% was higher than those from CP 18% group(P<0.05). Results from panel test showed that juiciness of meats from CP 16% group was higher than those from CP 12% group(P<0.05). Meats from CP 16% also was tested to be more tender than meats from the other groups. Results from this work suggest that the optimum crude protein level in growing goat's concentrate is 14-16% and that increase above this level seems not to improve meat production.

Development of K-bioassay for the Efficient Potassium Fertilization of Citrus Tree ($K(^{86}Rb)-bioassay$를 이용한 감귤나무의 가리영양진단법 개발)

  • U, Zang-Kual;Han, Hae-Ryong;Moon, Duk-Young;Kim, Chang-Myung;Lim, Han-Cheol;Moon, Do-Kyung;Song, Sung-Jun
    • Applied Biological Chemistry
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    • v.37 no.3
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    • pp.182-188
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    • 1994
  • Similar to the $^{42}K$ uptake, $^{86}Rb$ uptake by the roots of Hordeum distichum grown in the hydroponic culture was negatively correlated with the concentration of K supplied previously, showing that $^{86}Rb$ can be used for the K-bioassay. $^{86}Rb$ having longer half life (18.86 day) than $^{42}K$ (12.36 hr) allowed the use of larger number of root samples. $^{86}Rb$ uptake of 3 years old Citrus unshiu Marc. grown in water culture decreased drastically with the increase of K concentration of the culture solution, thus demonstrating that the nutrition status of K for citrus trees can be diagnosed by K-bioassay using $^{86}Rb$ tracer. $^{86}Rb$ uptake by the excised roots of Hordeum distichum grown in the pot with different K fertilizations was well correlated with the exchangeable K in soil. The amount of exchangeable K in soil for the optimal plant growth can be determined by its relationship. $^{42}K$ and $^{86}Rb-uptake$ by the Hordeum distichum roots were markedly inhibited by $5{\times}10^{-3}\; M$ KCN in the bioassay solution, indicating that uptake is energy-dependent. There was no significant relationship between K content in citrus leaves and K concentration in the water-culture medium. It is concluded that K-bioassay is a potentially useful tool for determining of K requirement in citrus trees.

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