• Title/Summary/Keyword: Comparison Research

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Biological Functions of N- and O-linked Oligosaccharides of Equine Chorionic Gonadotropin and Lutropin/Chorionic Gonadotropin Receptor

  • Min, K.S.
    • Korean Journal of Animal Reproduction
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    • v.24 no.4
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    • pp.357-364
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    • 2000
  • Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$ -subunits. The subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$ - and $\beta$-subunits in the placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t631 or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632~653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agoinst-occupied receptors ~2- and ~17- fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.

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A Comparison of Discriminating Powers Between 14 Microsatellite markers and 60 SNP Markers Applicable to the Cattle Identification Test (소 동일성 검사에 적용 가능한 14 Microsatellite marker와 60 Single Nucleotide Polymorphism marker 간의 판별 효율성 비교)

  • Lim, Hyun-Tae;Seo, Bo-Yeong;Jung, Eun-Ji;Yoo, Chae-Kyoung;Yoon, Du-Hak;Jeon, Jin-Tae
    • Journal of Animal Science and Technology
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    • v.51 no.5
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    • pp.353-360
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    • 2009
  • When 14 microsatellite (MS) markers were applied in the identifying test for 480 Hanwoo, the discriminating power was estimated as $3.43{\times}10^{-27}$ based on the assumption of a random mating group (PI). This rate is 1,000 times higher than that of 60 single nucleotide polymorphism (SNP) markers. On the other hand, the power of the 60 SNP markers was estimated as $4.69{\times}10^{-20}$ and $8.02{\times}10^{-12}$ on the assumption of a half-sib mating group ($PI_{half-sibs}$) and a full-sib mating group ($PI_{sibs}$), respectively. These powers were 10 times and 10,000 times higher than those of the 14 MS markers. The results indicated that the total number of alleles (MS vs SNP = 146 vs 120) acted as a key factor for the discriminating power in a random mating population, and the total number of markers (MS vs SNP = 14 vs 60) was a dominant influence on the power in half-sib and full-sib populations. In the Hanwoo population, in which it was assumed that the entire population is the enormous half-sib group formed by the absolute genetic contribution of a few nuclear bulls, there will be only a 10 times difference in the discriminating power between the 14 MS markers and the 60 SNP makers. However, the probability of not excluding a candidate parent pair from the parentage of an arbitrary offspring, given that only the genotype of the offspring ($PNE_{pp}$) was 1,000 times higher as shown by the 14 MS markers than that by the 60 SNP markers. The strong points of SNP makers are the stability of the variation (low mutation rate) and automation of high-throughput genotyping. In order to apply these merits for the practical and constant Hanwoo identity test, research and development are required to set a cost-effective platform and produce a homemade apparatus for SNP genotyping.

Analysis of Mistakes in Photosynthesis Unit in Biology II Textbooks and Survey of Biology Teachers' Recognition on them (생물 II 교과서 광합성 단원의 오류 분석 및 생물 교사의 오류 인지 조사)

  • Park, Hae-Kyung;Yoon, Ki-Soon;Kwon, Duck-Kee
    • Journal of Science Education
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    • v.32 no.1
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    • pp.33-46
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    • 2008
  • The purpose of this study was to know whether or not any wrongful description or simple errors were in photosynthesis unit of Biology II textbook under 7th national curriculum and if so, to know whether or not high school teachers recognized and corrected properly the mistakes. The mistakes in photosynthesis unit of text books were determined by the comparison with several reference books and through examination by three plant physiologists in 8 different Biology II textbooks. After the mistakes were analysed, the survey using contents of textbook containing the mistakes was conducted on high school teachers teaching Biology II. As a result, 48 mistakes were determined in 13 subjects. As many as four mistakes were found even in one subject in a certain textbook and a same mistake was found repeatedly in several textbooks. The survey result showed that the teachers who pointed exactly the mistakes out corrected properly, however, the percentage of these ones out of 35 teachers replied to survey was less than 50%. The ratios of correction out of total number of responses were high in question #6 (43%), #4-3 (40%), and #1-2 (32%) which were containing a simple mistake in graph, a wrong word and a wrong picture, respectively. But, no one pointed out and made correction in question #5-1 and #5-2 which were containing Z scheme of light reaction without the legend of vertical axis that should be explained as electron energy or standard reduction potential. The result indicates the possibility that the mistakes in photosynthesis unit of Biology II textbook can be corrected and teached properly by teachers may be low. In order to reduce the possibility that students may have misconceptions about photosynthesis, the list of print's errors should be provided to the teachers and/or the training program and/or workshop for in-service high school biology teachers was recommended.

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Studies on canine babesiosis in Korea I. In vitro isolation and antigenic properties of Babesia gibsoni (개 바베시아병에 관한 연구 I. Babesia gibsoni의 시험관내 분리와 항원성상에 관한 연구)

  • Lee, Ho-kweon;Suh, Myung-deuk
    • Korean Journal of Veterinary Research
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    • v.36 no.3
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    • pp.681-692
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    • 1996
  • The present study was conducted to isolate Babesia gibsoni by culture method of the microaerophilous stationary phase(MASP) and analyse the antigenic properties of the parasite by SDS-PAGE and immunoblot. The results obtained were summarized as follows. The protozoan parasite Babesia gibsoni multiplied in canine erythrocytes in RPMI 1640 medium(pH7.0) containing 20 40% normal canine serum under the MASP condition of 5% CO2 and 95% air at $37^{\circ}C$ incubator. The levels of parasitaemia in the erythrocytes were shown more higher by exchanging the medium at 24 hours interval. Under the above condition of MASP, the percentage of parasitized erythrocytes(PPE) after incubation for 8 days increased about 14 times more than that in the initiation of the 1% infected canine erythrocyte culture. The parasites were purely isolated from the MASP culture of red blood cells collected from dogs infected with Babesia gibsoni naturally or artificially. Among the total of 36 canine(Pit-bullterier) blood samples the parasites were isolated from 17 cases(47.2%) in the MASP culture while the parasites were detected from 20 cases(56%) and 12 cases(33.3%), respectively, by indirect fluorescent antibody(IFA) test and direct light microscopy(DLM). On the other hand, Babesia gibsoni was isolated by MASP culture from 15 cases(75%) and 11 cases(92%) of positive cases of IFA and DLM, respectively. In the analysis of the erythrocytic merozoite(AEOM) antigen derived from infected dog approximately 11 antigenic bands in molecular weight of 130, 120, 97.4, 92, 80, 52, 50, 42, 36, 30 and 29 KDa were observed on SDS-PAGE. Antigenic bands in the endoerythrocytic merozoite(CEOM) antigen derived from infected erythrocyte (sediment) in MASP culture were much similar to those of AEOM bands. In the exoerythrocytic merozoite(CEEM) antigen derived from supernatant of the infected erythrocyte culture approximately 20 antigenic bands were observed and the molecular weight of the major bands among these were 140, 120, 114, 105, 96, 93, 92, 80, 60, 52, 50, 38, 36, 30, 24, 18.5 and 16 KDa. In the protein patterns of AEOM and CEOM antigen by immunoblot 15 bands were observed and these patterns were much similar between each other. The molecular weight of the major bands in the both antigens were 130, 120, 80, 60, 52, 50, 42, 30, 29, 18.5 and 16 KDa. Approximately 21 bands were observed in CEEM antigen and the molecular weight of the major bands were 140, 120, 96, 92, 85, 80, 76, 60, 52, 50, 37, 30, 24, 16 and 15 KDa. The specific antigenic bands in the artificially infected dogs were firstly observed at 3 weeks afrer inoculation of infected blood and these antigenic bands were maintained up to 18 months after inoculation. In the immunoblot of the sera of the splenectomized dogs the specific antigenic bands with the molecular weight of 93 KDa and 52 KDa, respectively, were observed weakly comparing to those of non-splenectomized dog. In immunoblot of the sera collected from the naturally infected dogs the antigenic bands were observed as same as those of artificially infected dogs while antigenic band of 29 KDa in some individual dog showed strongly. In comparison of immunoblot of the sera collected from dogs non-treated and treated with diminazene aceturate(7mg/kg, IM) after artificial infection no differences of antigenic bands were observed. In analysis of antigenic bands by digoxigenin glycan/protein double labeling, antigenic bands in the molecular weight of 106, 60 58, 36, 30 and 29 KDa were determined as glycoproteins.

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Seasoning of Commercial Wood Using Solar Energy (태양에너지를 이용한 유용목재의 건조)

  • Jung, Hee-Suk;Lee, Hyoung-Woo;Lee, Nam-Ho;Lee, Sang-Bong
    • Journal of the Korean Wood Science and Technology
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    • v.16 no.4
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    • pp.10-39
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    • 1988
  • This study investigated the temperatures and relative humidities in the semi-greenhouse type solar dryer with a black rock-bed heat storage and without heat storage and outdoor temperature and relative humidity at 9 a.m. and 2 p.m.. A comparison was made of the drying rates, final moisture contents, moisture content distributions, casehardening stresses, drying defects, volumetric shrinkage of dried lumber for solar- and air-drying from the green condition of mixtures of Douglas-fir, lauan, taun, oak and sycamore 25mm- and 50 mm-thick lumber during the same period for four seasons, and heat efficiencies for solar dryer with and without the heat storage for saving of heat energy and the cost of lumber drying using the solar energy. The results from this study were summarized as follows: I. The mean weekly temperatures in the solar dryers were 3 to $6^{\circ}C$ at 9 a.m. and 9 to $13^{\circ}C$ at 2 p.m. higher than mean outdoor temperature during all the drying period. 2. The mean weekly relative humidities in the solar dryers were about 1 to 19% at 9 a.m. higher than the outdoor relative humidity. and the difference between indoor and outdoor relative humidity in the morning was greater than in the afternoon. 3. The temperatures and relative humidities in the solar dryer with and without the heat storage were nearly same. 4. The overall solar insolation during the spring months was highest and then was greater in the order of summer, atumm, and winter month. S. The initial rate of solar drying was more rapid than that of air drying. As moisture content decreased, solar drying rate became more rapid than that of air drying. The rates of solar drying with and without heat storage were nearly same. The drying rate of Douglas-fir was fastest and then faster in the order of sycamore, lauan, taun and oak. and the faster drying rate of species, the smaller differences of drying rates between thicknesses of lumber. The drying rates were fastest in the summer and slowest in the winter. The rates of solar drying during the spring were more slowly in the early stage and faster in the later stage than those during the autumn. 6. The final moisture contents were above 15% for 25mm-thick air dried and about 10% for solar dried lumber, but the mean final MCs for 50mm-thick lumber were much higher than those of thin lumber. The differences of final MC between upper and lower course of pile for solar drying were greater than those of pile for air drying. The differences of moisture content between the shell and the core of air dried lumbers were greater than those of solar dried lumber, smallest in the drying during summer and greatest in the drying during winter among seasons. 7. Casehardening stresses of 25mm- and 50mm-thick dried lumber were slight, casehardening stress of solar dried lumber was severer than that of air dried lumber and was similar between solar dried lumber with and without heat storage, Casehardening stresses of lumber dried during spring were slightest and then slighter in the order of summer, autumn, and winter. Casehardening stresses of Douglas -fir, sycamore and lauan were slight, comparing with those of taun and oak. 8. Maximum initial checks of 25mm-thick lumber occurred above and below fiber saturation point and those of 50mm-thick lumber occurred in the higher moisture content than thin lumber. As the moisture content decreased, most of checks were closed and didn't show distinct difference of the degree of checks among drying methods. The degree of checks were very slight in case of Douglas-fir and lauan, and severe in case of taun and oak. The degree of checks for 50mm-thick lumber were severer than those for 25mm-thick lumber. 9. The degree of warpage showed severe in case of oak and sycamore lumber, but no warping was found in case of Douglas-fir, lauan and taun. 10. The volumetric shrinkages of taun and oak were large and medium in case of Douglas-fir, lauan and sycamore. 11. Heat efficiencies of solar dryer with heat storage were 6.9% during spring, 7.7% during summer, 12.1% during autumn and 4.1% during winter season. Heat efficiency of solar dryer with heat storage was slightly greater than that of without heat storage. As moisture content of lumber decreased, heat efficiency decreased.

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The Evaluation of Reconstructed Images in 3D OSEM According to Iteration and Subset Number (3D OSEM 재구성 법에서 반복연산(Iteration) 횟수와 부분집합(Subset) 개수 변경에 따른 영상의 질 평가)

  • Kim, Dong-Seok;Kim, Seong-Hwan;Shim, Dong-Oh;Yoo, Hee-Jae
    • The Korean Journal of Nuclear Medicine Technology
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    • v.15 no.1
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    • pp.17-24
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    • 2011
  • Purpose: Presently in the nuclear medicine field, the high-speed image reconstruction algorithm like the OSEM algorithm is widely used as the alternative of the filtered back projection method due to the rapid development and application of the digital computer. There is no to relate and if it applies the optimal parameter be clearly determined. In this research, the quality change of the Jaszczak phantom experiment and brain SPECT patient data according to the iteration times and subset number change try to be been put through and analyzed in 3D OSEM reconstruction method of applying 3D beam modeling. Materials and Methods: Patient data from August, 2010 studied and analyzed against 5 patients implementing the brain SPECT until september, 2010 in the nuclear medicine department of ASAN medical center. The phantom image used the mixed Jaszczak phantom equally and obtained the water and 99mTc (500 MBq) in the dual head gamma camera Symbia T2 of Siemens. When reconstructing each image altogether with patient data and phantom data, we changed iteration number as 1, 4, 8, 12, 24 and 30 times and subset number as 2, 4, 8, 16 and 32 times. We reconstructed in reconstructed each image, the variation coefficient for guessing about noise of images and image contrast, FWHM were produced and compared. Results: In patients and phantom experiment data, a contrast and spatial resolution of an image showed the tendency to increase linearly altogether according to the increment of the iteration times and subset number but the variation coefficient did not show the tendency to be improved according to the increase of two parameters. In the comparison according to the scan time, the image contrast and FWHM showed altogether the result of being linearly improved according to the iteration times and subset number increase in projection per 10, 20 and 30 second image but the variation coefficient did not show the tendency to be improved. Conclusion: The linear relationship of the image contrast improved in 3D OSEM reconstruction method image of applying 3D beam modeling through this experiment like the existing 1D and 2D OSEM reconfiguration method according to the iteration times and subset number increase could be confirmed. However, this is simple phantom experiment and the result of obtaining by the some patients limited range and the various variables can be existed. So for generalizing this based on this results of this experiment, there is the excessiveness and the evaluation about 3D OSEM reconfiguration method should be additionally made through experiments after this.

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Functional Properties of Walnut in Cosmetics (호두의 화장품 기능성)

  • Kim, Mee-Kyung;Kim, Jin-Sung;Jo, Bun-Sung;Kim, Jeung-Hoan;Lee, In-Cheol;Lee, Myung-Sup;Cho, Young-Je
    • Journal of Life Science
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    • v.21 no.6
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    • pp.858-864
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    • 2011
  • The phenolic compounds of walnut extracts by various solvents were shown to be 24.3 mg/g in hot water, 34.4 mg/g in ethanol, 32.5 mg/g in methanol and 15.1 mg/g in acetone. In a comparison of phenolic compounds from hot water and different concentrations of ethanol, which are harmless, 60% ethanol extract and hot water extract were 34.7 mg/g, 24.6 mg/g. The electron donating ability (EDA) of walnut extracts in hot water and 60% ethanol were 78.1% and 80.6%. According to ABTS radical cation decolorization for antioxidant activity, hot water and 60% ethanol extract showed high antioxidant activities of 98.1% and 98.3%. Antioxidant protection factor (PF) were $1.1{\pm}0.2$ PF and $1.1{\pm}0.4$ PF in hot water and 60% ethanol extract. In TBARs inhibitory activity, each extract showed high antioxidant activities at 60% and 75%. Anti-inflammation effects of walnut extract were tested, and inhibition of NO was 50% in 100 ${\mu}g/ml$ phenolics. Inhibitory activity against iNOS and COX-2 were shown, through Western blot, to be 10% in 100 ${\mu}g/ml$ phenolics. Tyrosine inhibitory activity of 60% ethanol extract was 43%, and astringent effect of 60% ethanol extract was 55%. These results suggest that walnut extracts are suitable for functional cosmetics requiring skin-whitening and anti-wrinkle activity.

Effect of Yearly Application of Straw and Nitrogen, Phosphate and Potassium Fertilizer on the Growth and Yield in Rice (생고(生藁) 및 삼요소(三要素)의 연속시용(連續施用)이 수도(水稻)의 생육(生育) 및 수량(收量)에 미치는 영향(影響))

  • Lee, Eun-Woong;Kwon, Y.W.
    • Applied Biological Chemistry
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    • v.8
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    • pp.51-59
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    • 1967
  • A split plot was laid out in 1965 for perennial application of straw by 500kg per 10 are, as main plot, and of chemical fertilizer in conventional way, as sub-plots. The sub-plot included the following 5 fertilizer treatments as (1) non-fertilized, (2) nitrogen-free, (3) phosphate-free (4) potassium-free, (5) completely fertilizer applied plots. Trends in the growth and yield of rice was observed at present experimental site in the first year, 1965, and second year, 1966, as follows: 1. Each treatmental plot showed nearly identical trends between the first and second year, in term of growth, yield and yield components. 2. Straw application deterred the initial growth at non-fertilized and nitrogen-free plots. But straw application raised the effective stem ratio and enlarged the values in yield components, finally to be resulted in the second year with significant difference. 3. Both phosphate and potassium did not much affected to plants for longer stem and larger number of tillers. 4. Effect of nitrogen, phosphate and potassium was recorded as investigated by previous research works. Phospate deficiency decreased maturing rate to a great extent. 5. Numerical index was calculated for the comparison of plots in yield, where the non-straw, completely fertilizer applied plot stands for 100. The indexes of the first and second year were as follows : at non-straw plot(indexes in the blank are from the first year trial); non-fertilized : 80.2(80.9), nitrogen-free: 83.6(89.4), phosphate-free : 89.4(93.1) and potassium-free plot: 93.5(102.4) at non-straw plot. On the other hand, indexes at straw applied plots were non-fertilized : 84.0(86.6), nitrogen-free 82.6(93.9), phosphate-free : 91.7(96.3), potassium-free: 103.2(102.0) and complete plot: 98.7(109.8).

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Screening for Effective Organic Farming Materials for the Control of Cucumber Scab Caused by Cladosporium cucumerinum (오이 검은별무늬병 방제에 효과적인 유기농업자재 선발)

  • Park, So-Hyang;Hong, Sung-Jun;Shim, Chang-Ki;Kim, Min-Jeong;Park, Jong-Ho;Han, Eun-Jung;Kim, Yong-Ki
    • Research in Plant Disease
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    • v.23 no.2
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    • pp.159-167
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    • 2017
  • This study was conducted to evaluate the control efficacy of the organic farming materials on cucumber scab caused by Cladosporium cucumerinum PT1 (KACC 48094). The antifungal activities in vitro as well as the suppressive effect of 43 organic farming materials on the spore germination and germ tube growth by inoculating spore suspension on cucumber seedlings in vivo were investigated. Thirteen organic farming materials inhibited the mycelial growth of C. cucumerinum and nine of these were microbial agents. In the screening using cucumber seedlings, six organic farming materials were very effective with control efficacy value of 90%. Among them, Bacillus amyloliquefaciens M27 provided suppressive effect on both mycelial growth and spore germination against cucumber scab. Finally, nine organic farming materials were selected to test the protective and curative effects, and all chosen organic farming materials significantly suppressed disease incidence when applied in the preventive action, in comparison with the curative action. Especially, Bordeaux mixture I and III gave excellent protective control efficacy with control values of 96.7% and 73.3%, respectively, whereas its curative control effect was significant low. Among these, only Thymus quinquecostatus+Sophora extract showed curative activity, although the control value was as low as 50%. This study suggests that cucumber scab can be controlled by some organic farming materials in the farmhouses under comparatively cold and wet condition and protective treatment is more important and efficient.

Evaluation of the Temperature Drop Effect and the Rutting Resistance of Moisture Retaining-Porous Asphalt Pavement Using Accelerated Pavement Testing (포장가속시험을 이용한 보수형 배수성 포장의 온도저감 효과 및 소성변형 저항특성 연구)

  • Kwak, Byoung-Seok;Suh, Young-Chan;Song, Chul-Young;Kim, Ju-Won
    • International Journal of Highway Engineering
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    • v.11 no.3
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    • pp.97-109
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    • 2009
  • One of the main causes of asphalt rutting is high temperature of the pavement. Nevertheless, there has been few research on lowering the pavement temperature for reducing rutting. This study investigated the performance characteristics of moisture-retaining porous asphalt pavement, which is known to have a temperature reducing effect. The purpose of this study is to quantify the temperature reducing effect of moisture-retaining porous asphalt pavement and its effect of reducing rutting through Accelerated Pavement Testing(APT). Additionally, the possibility of reducing the thickness of the pavement in comparison to general dense grade pavement by analyzing structural layer coefficient of moisture retaining pavement. A total of three test sections consisting of two moisture-retaining porous asphalt pavement sections and one general dense-grade porous asphalt pavement section were constructed for this study. Heating and spraying of water were carried out in a regular cycle. The loading condition was 8.2 ton of wheel load, the tire pressure of $7.03kgf/cm^2$, and the contact area of $610cm^2$. The result of this experiment revealed that the temperature reducing effect of the pavement was about $6.6{\sim}7.9^{\circ}C$(average of $7.4^{\circ}C$) for the middle layer and $7.9{\sim}9.8^{\circ}C$(average of $8.8^{\circ}C$) for surface course, resulting in a rutting reduction of 26% at the pavement surface. Additionally, the structural layer coefficient of moisture retaining pavement measured from a laboratory test was 0.173, about 1.2 times that of general dense grade pavement. The general dense-grade porous asphalt pavement test section exhibited rutting at all layers of surface course, middle layer, and base layer, while the test sections of moisture-retaining porous asphalt pavement manifested rutting mostly at surface course only.

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