• Title/Summary/Keyword: activity quantity

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Comparison of Total RNA Isolation Methods for Analysis of Immune-Related microRNAs in Market Milks

  • Oh, Sangnam;Park, Mi Ri;Son, Seok Jun;Kim, Younghoon
    • Food Science of Animal Resources
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    • v.35 no.4
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    • pp.459-465
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    • 2015
  • Bovine milk provides essential nutrients, including immunologically important molecules, as the primary source of nutrition to newborns. Recent studies showed that RNAs from bovine milk contain immune-related microRNAs (miRNA) that regulate various immune systems. To evaluate the biological and immunological activity of miRNAs from milk products, isolation methods need to be established. Six methods for extracting total RNAs from bovine colostrums were adopted to evaluate the isolating efficiency and expression of miRNAs. Total RNA from milk was presented in formulation of small RNAs, rather than ribosomal RNAs. Column-combined phenol isolating methods showed high recovery of total RNAs, especially the commercial columns for biofluid samples, which demonstrated outstanding efficiency for recovering miRNAs. We also evaluated the quantity of five immune-related miRNAs (miR-93, miR-106a, miR-155, miR-181a, miR-451) in milk processed by temperature treatments including low temperature for long time (LTLT, 63℃ for 30 min)-, high temperature for short time (HTST, 75℃ for 15 s)-, and ultra heat treatment (UHT, 120-130℃ for 0.5-4 s). All targeted miRNAs had significantly reduced levels in processed milks compared to colostrum and raw mature milk. Interestingly, the amount of immune-related miRNAs from HTST milk was more resistant than those of LTLT and UHT milks. Our present study examined defined methods of RNA isolation and quantification of immune-specific miRNAs from small volumes of milk for use in further analysis.

Identification of a Novel Single Nucleotide Polymorphism in Porcine Beta-Defensin-1 Gene

  • Pruthviraj, D.R.;Usha, A.P.;Venkatachalapathy, R.T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.3
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    • pp.315-320
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    • 2016
  • Porcine beta-defensin-1 (PBD-1) gene plays an important role in the innate immunity of pigs. The peptide encoded by this gene is an antimicrobial peptide that has direct activity against a wide range of microbes. This peptide is involved in the co-creation of an antimicrobial barrier in the oral cavity of pigs. The objective of the present study was to detect polymorphisms, if any, in exon-1 and exon-2 regions of PBD-1 gene in Large White Yorkshire (LWY) and native Ankamali pigs of Kerala, India. Blood samples were collected from 100 pigs and genomic DNA was isolated using phenol chloroform method. The quantity of DNA was assessed in a spectrophotometer and quality by gel electrophoresis. Exon-1 and exon-2 regions of PBD-1 gene were amplified by polymerase chain reaction (PCR) and the products were subjected to single strand conformation polymorphism (SSCP) analysis. Subsequent silver staining of the polyacrylamide gels revealed three unique SSCP banding patterns in each of the two exons. The presence of single nucleotide polymorphisms (SNPs) was confirmed by nucleotide sequencing of the PCR products. A novel SNP was found in the 5'-UTR region of exon-1 and a SNP was detected in the mature peptide coding region of exon-2. In exon-1, the pooled population frequencies of GG, GT, and TT genotypes were 0.67, 0.30, and 0.03, respectively. GG genotype was predominant in both the breeds whereas TT genotype was not detected in LWY breed. Similarly, in exon-2, the pooled population frequencies of AA, AG, and GG genotypes were 0.50, 0.27, and 0.23, respectively. AA genotype was predominant in LWY pigs whereas GG genotype was predominant in native pigs. These results suggest that there exists a considerable genetic variation at PBD-1 locus and further association studies may help in development of a PCR based genotyping test to select pigs with better immunity.

Genotoxicity on Structural Derivatives of Sophoricoside, a Component of Sophora Japonica, in Bacterial and Mammalian Cells

  • Ryu, Jae-Chun;Kim, Youn-Jung;Kim, Mi-Soon;Kim, Min-Ji;Sarma, Sailendra Nath;Jung, Sang-Hun
    • Molecular & Cellular Toxicology
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    • v.1 no.3
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    • pp.179-188
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    • 2005
  • To develop the novel anti-allergic drug, many sophoricoside derivatives were synthesized. Among these derivatives, JSH-II-3, VI-3, VII-3, VIII-3, VII-20 and VII-20 (sodium salt) were selected and subjected to high throughput toxicity screening (HTTS) because they revealed strong IL-5 inhibitory activity and limitation of quantity. Single cell gel electrophoresis (Comet) assay, mouse lymphoma thymidine kinase ($tk^{+/-}$) gene assay (MOLY), chromosomal aberration assay in mammalian cells and Ames reverse mutation assay in bacterial system were used as simplified, inexpensive, short-term in vitro screening tests in our laboratory. Through the primary screening using the comet assay, we could choose the first candidates of sophoricoside derivatives with no genotoxic potentials as JSH-VI-3, VII-3, VII-20 and VII-20 (sodium salt). Also JSH-VII-3, VII-20 and VII-20 (sodium salt) are non-mutagenic in MOLY assay, while JSH-II-3 is mutagenic at high concentration with the presence of metabolic activation system in both comet assay and MOLY assay. The selected derivatives (JSH-VI-3, VII-3, VII-20 and VII-20 (sodium salt) are not mutagenic in S. typhimurium TA98 and TA100 strains both in the presence and absence of metabolic activation. From results of chromosomal aberration assay, 6 h treatment of JSH-VI-3, VII-3 and VII-20 (sodium salt) were not revealed clastogenicity both in the presence and absence of S-9 mixture. Therefore, we suggests that JSH-VI-3, VII-3, VII-20 and VII-20 (sodium salt), as the optimal candidates with both no genotoxic potential and IL-5 inhibitory effects must be chosen. To process the development into new anti-inflammatory drug of these derivatives, further investigation will need.

Quality Characteristics of $Sikhea$ with Mulberry Fruit (오디 첨가한 식혜의 품질특성에 관한 연구)

  • Kim, Jung-Soo
    • Culinary science and hospitality research
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    • v.18 no.2
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    • pp.206-215
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    • 2012
  • The results of influence on the saccharification and $Sikhea$ by adding mulberry fruit in this study are as follows. pH was decreased with the amount of mulberry fruit increased during the saccharification period. A significant change was not found after 4 hours of saccharification. It is considered that 4-5 hours was proper for saccharification because brix increased by more than 1 brix per hour up to 4 hours while there were little change after 5 hours. L was decreased while a and b values were increased in the Hunter' value of mulberry fruit $Sikhea$. The DPPH free radical scavenging activities and SOD-like activity of mulberry fruit $Sikhea$ were increased as the addition quantity of mulberry fruit increased. Anthocynin color pigment in mulberry fruit has the inhibiting effect on microbes. The preferences of mulberry fruit $Sikhea$ were 10% < 30% < 15% < 20% < 0% < 25%, and less than 25% of mulberry fruit addition seems to be desirable. The addition of mulberry fruit will help decrease the amount of sugar to use in $Sikhea$ because the sugar level of $Sikhea$ increases with the addition of mulberry fruit. It will also improve the preference for Sihkea by giving it better color and sourness.

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벙커-C유 분해미생물의 선발 및 현장 생리적용성 검정

  • Park, Jin-Hui;Kim, Yeong-Sik;Lee, Tae-Jin
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.600-604
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    • 2003
  • This study pointed at isolation of bunker-C oil degrading bacteria and then estimation of it's degrading capability in environmental conditions. Degradation ratio of the excellent isolate was appeared to 40.5% and 44.7% when the oil was treated to 1% and 5%, respectively. The isolate was identified to Acinetobater calcoaceticus SEBCM. In pH test, high degrading effect was appeared to about 73% at pH 6 and pH 7, and low degrading ratio was 37% at pH 4. Its growth condition at temperature has not large variation in $15^{\circ}C\;{\sim}30^{\circ}C$, Quantity of nitrogen for it's good growth was ranged of $0.5\;g/L{\sim}2\;g/L$. As these results, we realized that this isolate have good activity when treated to $15\;{\sim}30^{\circ}C$ of temperature and $6{\sim}7$ of pH.

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The Variation of Surface Area in Porous Poly(Styrene-co-Divinylbenzene) Resin Beads (다공성 스티렌-디비닐벤젠 공중합 수지입자의 표면적 변화)

  • Kim, Yong-Man;Ihm, Son-Ki;Kim, Jong-Chan;Lee, Dong-Keun;Ahn, Jou-Hyeon
    • Applied Chemistry for Engineering
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    • v.7 no.5
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    • pp.843-848
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    • 1996
  • Porous resin beads of Poly(styrene-co-divinylbenzene) have been prepared by suspension polymerization. The bead could be made porous in the region above 30wt% of the crosslinking agent(divinylbenzene ) and the porogenic agent(toluene), respectively. The specific surface area of porous beads increased with increasing the concentrations of divinylbenzene and toluene. The specific surface area of the porous resin bead decreased, when sulfonated with concentrated sulfuric acid. The catalytic activity of sulfonated resin catalyses increased with increasing the degree of crosslinking in the liquid-phase reesterification of ethyl acetate with 1-propanol. The adsorbed quantity of sodium dodecylbenzene sulfonate in an aqueous solution also increased with increasing surface area of porous resins.

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Influences of Mixing Frequency on the Composting Performance of the Broiler and Dairy Manure Mixtures (퇴비재료의 교반빈도가 육계분과 유우분 혼합물 퇴비화 성능에 미치는 영향)

  • Park K.J.;Bae Y.H.;Hong J.H.;Wi T.W.
    • Journal of Animal Environmental Science
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    • v.12 no.1
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    • pp.41-44
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    • 2006
  • Broiler manure has much nutrient that can be used as the organic fertilizer to enhance the fertility of soil. However, if it is used directly without biodegradation of organic materials and destruction of weed seed and harmful bacteria, it can produce the generation of weed and disease of plant. Composting of manure is a biodegradation of organic materials into inorganic materials and humus. To proceed biodegradation of manure effectively and enhance the composting performance, optimum environmental condition for microbial growing should be maintained. Environmental variables which can influence the growing activity of microbes are moisture content, pH, porosity, C/N of the composting materials and oxygen supply quantity. Oxygen and porosity are usually supplied by aeration or mixing of materials. This study was intended to investigate the effect of mixing frequency on the composting performance. Mixing of composting materials was performed by turning the bioreactor up and down by hand without any mechanical energy. The broiler manure was captured from the greenhouse type broiler ham as the compounds of broiler manure and rice-hulls, which were used as the base materials. To compost the compounds of broiler manure and rice-hulls, dairy manure was mixed to get appropriate characteristics of composting material. Composting temperature over $55^{\circ}C$ for killing pathogen and weed seed was maintained for longer period with increase of mixing frequency.

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A study on an intermittent aeration membrane bioreactor system using ammonia sensor to decrease energy consumption and sludge concentration by tubular membrane (암모니아 센서를 이용한 간헐폭기 Membrane bioreactor공정에서의 전력비 저감과 관형막을 이용한 슬러지 농축에 관한 연구)

  • Kang, Heeseok;Lee, Euijong;Kim, Hyungsoo;Jang, Am
    • Journal of Korean Society of Water and Wastewater
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    • v.28 no.2
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    • pp.161-170
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    • 2014
  • It is essential to decrease energy consumption and excess sludge to economically operate sewage treatment plant. This becomes more important along with a ban on sea dumping and exhaustion of resource. Therefore, many researchers have been study on energy consumption reduction and strategies for minimization of excess sludge production from the activated sludge process. The aeration cost account for a high proportion of maintenance cost because sufficient air is necessary to keep nitrifying bacteria activity of which the oxygen affinity is inferior to that of heterotrophic bacteria. Also, additional costs are incurred to stabilize excess sludge and decrease the volume of sludge. There were anoxic, aerobic, membrane, deairation and concentration zone in this MBR process. Continuous aeration was provided to prevent membrane fouling in membrane zone and intermittent aeration was provided in aerobic zone through ammonia sensor. So, there was the minimum oxygen to remove $NH_4-N$ below limited quantity that could be eliminated in membrane zone. As the result of this control, energy consumption of aeration system declined by between 10.4 % and 19.1 %. Besides, we could maintain high MLSS concentration in concentration zone and this induced the microorganisms to be in starved condition. Consequentially, the amount of excess sludge decrease by about 15 %.

Ex vivo Cytotoxicity of the Bacillus thuringiensis Cry4B δ-Endotoxin to Isolated Midguts of Aedes aegypti Larvae

  • Barusrux, Sahawat;Sramala, Issara;Katzenmeier, Gerd;Bunyaratvej, Ahnond;Panyim, Sakol;Angsuthanasombat, Chanan
    • BMB Reports
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    • v.36 no.3
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    • pp.294-298
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    • 2003
  • The pathological effect of the Bacillus thuringiensis Cry $\delta$-endotoxins on susceptible insect larvae had extensive damage on the midgut epithelial cells. In this study, an ex vivo assay was devised for assessing the insecticidal potency of the cloned Cry4B mosquito-larvicidal protein that is expressed in Escherichia coli. Determination of toxicity was carried out by using a cell viability assay on the midguts that were dissected from 5-day old Aedes aegypti mosquito larvae. After incubation with the toxin proteins, the number of viable epithelial cells was determined photometrically by monitoring the quantity of the bioreduced formazan product at 490 nm. The results showed that the 65-kDa trypsin-activated Cry4B toxin exhibited toxic potency ca. 3.5 times higher than the 130-kDa Cry4B protoxin. However, the trypsin-treated products of the non-bioactive Cry4B mutant (R158A) and the lepidopteran-specific Cry1Aa toxin displayed relatively no ex vivo activity on the mosquito-larval midguts. The ex vivo cytotoxicity studies presented here confirms data that was obtained in bioassays.

A Study on Public Needs for Privately Owned Public Space (실내공적공간의 공공성에 관한 연구)

  • Yun, Ji-Hye;Kim, Jung-Gon
    • Korean Institute of Interior Design Journal
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    • v.15 no.5 s.58
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    • pp.157-166
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    • 2006
  • Recently, it appears several counterproposals about desirable figures of urban architecture. All of them proposes 'publicity' with cohernt tendency. The reason why it concentrates quantitative expansion of city without united design by urban plannar is that neglect quality values of city. As a solution of poor environment, there cue out the various efforts, about problem of each building, problem of city space, problem of laws and so forth. The reason why necessity of public space was embossed in that architecture extend the activity of citizen and make up the city space. But, each building pursues the private interest, so it is difficult to secure a public space with a high hand. Thus, architecture law has been revised in 1991 and bring the system of open space to match up the publicity and the private interest. Actually, western country brought it and obtained excellent results. While quantity of open space have increased since 1991, a lot of problems revealed in real usage and quality. By means of problem's solution, this study focus on the diversion of recognition for necessity of various open space. In result, on the occasion of approach and openess, except for several building, most glass a facade and the pedestrian can approach easily. Moreover, office buildings near the subway station connected with their low floor. So, the office buildings give openess to pedestrian and a people can approach easily to the buildings. On the occasion of amenity, most have bank and lobby on the first floor and have facilities on the underground floor. It leave open. But the reason why they have bank and lobby is that the space is dry and boring(without elements of nature and rest space). Hence, to make a space full of vitality, it have to plan various design elements and facilities. First of all, plan of indoor public space have to make up facility for the public interest. This study is basic investigation for necessity of indoor public space and through the survey of office buildings, it analyze the character of plan and find out the method of publicity's realization.