• Title/Summary/Keyword: Bio membrane

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Optimization of Protoplast Isolation and Ribonucleoprotein/Nanoparticle Complex Formation in Lentinula edodes (표고버섯의 원형질체 분리 최적화와 RNPs/나노파티클 복합체 형성)

  • Kim, Minseek;Ryu, Hojin;Oh, Min Ji;Im, Ji-Hoon;Lee, Jong-Won;Oh, Youn-Lee
    • Journal of Mushroom
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    • v.20 no.3
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    • pp.178-182
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    • 2022
  • Despite the long history of mushroom use, studies examining the genetic function of mushrooms and the development of new varieties via bio-molecular methods are significantly lacking compared to those examining other organisms. However, owing to recent developments, attempts have been made to use a novel gene-editing technique involving CRISPR/Cas9 technology and genetic scissors in mushroom studies. In particular, research is actively being conducted to utilize ribonucleoprotein particles (RNPs) that can be genetically edited with high efficiency without foreign gene insertion for ease of selection. However, RNPs are too large for Cas9 protein to pass through the cell membrane of the protoplasmic reticulum. Furthermore, guide RNA is unstable and can be easily decomposed, which remarkably affects gene editing efficiency. In this study, nanoparticles were used to mitigate the shortcomings of RNP-based gene editing techniques and to obtain transformants stably. We used Lentinula edodes (shiitake mushroom) Sanjo705-13 monokaryon strain, which has been successfully used in previous genome editing experiments. To identify a suitable osmotic buffer for the isolation of protoplast, 0.6 M and 1.2 M sucrose, mannitol, sorbitol, and KCl were treated, respectively. In addition, with various nanoparticle-forming materials, experiments were conducted to confirm genome editing efficiency via the formation of nanoparticles with calcium phosphate (CaP), which can be bound to Cas9 protein without any additional amino acid modification. RNPs/NP complex was successfully formed and protected nuclease activity with nucleotide sequence specificity.

Embryonic and Larval Development of Goby Minnow, Pseudogobio esocinus (모래무지(Pseudogobio esocinus)의 난발생 및 자치어 형태발달)

  • Lee, Sung-Hun;Oh, Gwang-Nam;Kim, Kwan-Seok;Oh, Yong-Seok;Kang, Kyeong-Wan;Hwang, Jae-Ho;Lee, Bae-ik;Lee, Won-Kyo;Han, Kyeong-Ho
    • Development and Reproduction
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    • v.12 no.3
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    • pp.283-288
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    • 2008
  • The Pseudogobio esocinus were caught at Wyuleo-ri, Gyeombaek-myeon, Boseong-gun, Jeollanamdo from April to May 2003. The fishes were incubated in transparent aquarium located at the laboratory of Chonnam National University, and their embryonic and larval development were observed. The fertilized eggs were spherical, semitransparent, and adhesive, and were $1.98{\pm}0.19mm$ (n=50) in diameter. The embryo, including 31$\sim$32 myotomes, hatched through egg membrane at 164 hrs after fertilization. The newly-hatched larvae were $4.61{\pm}0.83mm$ (n=10) in total length (TL). At that moment, yolk was not absorbed, and mouth and anus were not open. Star and spot shaped melanophores were distributed on the lens, and dorsal, ventral, and caudal parts. At 42 days after hatching, larva was $16.22{\pm}0.65mm$ (n=10) in TL. Melanophores were scaterred at head, back, and side parts. Morphological features of the embryo were transferred to juvenile stage showing similar features with those of the adult fish.

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Study on Anti-Helicobacter pylori Antibody of Sparated Antigen from H. pylori (Helicobacter pylori로부터 유래된 항원의 anti-H, pylori 항체에 관한 연구)

  • Park, Chang-Ho;Bae, Man-Jong
    • Journal of Life Science
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    • v.18 no.2
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    • pp.241-248
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    • 2008
  • This study has been carried out to secretion antibodies for the purpose of preventing the infection of Helicobacter pylori and using them as a supplement for treatment. This experiments have been separated antigens from H. pylori and observed into antibody production and the agglutination of H. pylori for the separated antigens. As major antigenic proteins separated from H. pylori, the following could be verified: 12 kinds of band for whole cell (WC), seven kinds of band for outer membrane protein (OMP), three kinds of band for crude urease, and one kind of band for lipopolysaccharide (LPS). The IgG anti-H. pylori antibody of separated antigens showed $77.9{\pm}6.4{\mu}g/ml$ for we (L), $84.9{\pm}6.4{\mu}g/ml$ for OMP, and $123.8{\pm}2.9{\mu}g/ml$ for crude urease, at the same antigen concentration of $20{\mu}g/100ull$, which showed the most at the crude urease. And it turned out that the IgA antibodies were generated with $2.5{\pm}0.32{\mu}g/ml$ for WC (L), $2.0{\pm}0.43{\mu}g/ml$ for OMP, and $1.3{\pm}0.25{\mu}g/ml$ for crude urease, which demonstrated the most for WC (L) antigens. As a result of verifying the immunogenecity of antigenic protein through the Western blotting, major antigenic substances could be confirmed as follows: 10 kinds for WC, six kinds for OMP and three kinds for crude urease. The agglutination values on the H. pylori of the antibody were $2^5,\;2^5,\;2^6\;and\;2^7$ at the antigen serums of anti-WC (H), anti-WC (L), anti-OMP and anti-crude urease, respectively, which indicated the highest for the antigen serum of anti-crude urease. The urease activation-inhibiting absorbance of antigen serum created by each antigen was $0.14{\pm}0.01$ for WC (H), $0.16{\pm}0.01$ for WC (L), $0.18{\pm}0.03$ for OMP, and $0.18{\pm}0.04$ for urease, demonstrating a significant inhibiting effect, compared with $0.26{\pm}0.02$ of the control group.

Effects of Concentrated Pig Slurry Using Membrane Filter on the Growth and Yield of Tomato in Nutriculture (막분리 돈분뇨 농축액비를 이용한 양액재배가 토마토의 생육과 수량에 미치는 영향)

  • Ryoo, Jong-Won;Seo, Woon-Kab
    • Journal of Animal Environmental Science
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    • v.14 no.2
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    • pp.119-128
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    • 2008
  • This experiment was conducted to investigate the effects of concentrated pig slurry using membrane filter on growth of tomato in nutriculture. Pig slurry was filtered by ultra filtration and concentrated by reverse osmosis process. Filtration of pig slurry was necessary to prevent the hose clogging in nutriculture. The concentrated pig slurry (CS) and nutrient solution (NS) were mixed by six different mixing ratios of 0:100, 20:80, 40:60, 60:40%, 80: 20 and 100%:0% based on nitrogen content. The chemical nutrient solution was the solution of National Horticulture Research Station for the growth of tomato. The concentration of nutrient solution was adjusted a range of $1.6{\sim}2.0mS/cm$ in EC. The plant height of tomato treated with CS 20+NS 80% was similar with NS 100% control plot. Plant height was highest in the plot of CS 20+NS 80%. The treatment of 100% concentrated pig slurry was lowest in the gowth characteristics of tomato. Number of cluster was very lower in 100% concentrated pig slurry compared with plot of chemical nutrient solution. In the beginning of growth stage, SPAD reading value was reduced in plot treated with CS 100%, but CS 20+NS 80% plot was higher compared to 100% concentrated pig slurry. SPAD value of tomato leaves was decreased as the amount of CS was increased. The SPAD value also in treatment of concentrated pig slurry was lower in the middle growth stage compared to control plot. The dry weight of stem and leaf were 107.4, 104.2g in plot of NS 100% and CS 20%+NS 80%, respectively. The fruit number and weight were decreased at high application plots of concentrated pig slurry, The fruit setting of tomato showed lowest in the plot treated with 100% concentrated pig slurry, and the growth of tomato severely decreased after application of 100% CS treatment. In conclusion, the growth characteristics such as plant height and fruit weight of tomato were not significantly different between the plots treated with mixture of 20% CS +80%NS and 100% nutrient solution treatment. In conclusion, the mixture solution of 20% of concentrated pig slurry and 80% of nutrient solution could be used as a nutrition solution of tomato nuticulture.

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Performance Evaluation of Bio-Membrane Hybrid Process for Treatment of Food Waste Leachate (음식물 침출수 청정화를 위한 파일롯 규모의 생물-분리막 복합공정의 성능 평가 연구)

  • Lee, Myung-Gu;Park, Chul-Hwan;Lee, Do-Hoon;Kim, Tak-Hyun;Lee, Byung-Hwan;Lee, Jin-Won;Kim, Sang-Yong
    • KSBB Journal
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    • v.23 no.1
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    • pp.90-95
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    • 2008
  • In this study, a combined process of sequential anaerobic-aerobic digestion (SAAD), fluidized-bed bioreactor (FBBR), and ultrafiltration (UF) for the treatment of small scale food waste leachate was developed and evaluated. The SAAD process was tested for performance and stability by subjecting leachate from food waste to a two-phase anaerobic digestion. The main process used FBBR composed of aerators for oxygen supply and fluidization, three 5 ton reaction chambers containing an aerobic mesophilic microorganism immobilized in PE (polyethylene), and a sedimentation chamber. The HRTs (hydraulic retention time) of the combined SAAD-FBBR-UF process were 30, 7, and 1 day, and the operation temperature was set to the optimal one for microbial growth. The pilot process maintained its performance even when the CODcr of input leachate fluctuated largely. During the operation, average CODcr, TKN, TP, and salt of the effluent were 1,207mg/L, 100mg/L, 50 mg/L, and 0.01 %, which corresponded to the removal efficiencies of 99.4%, 98.6%, 89.6%, and 98.5%, respectively. These results show that the developed process is able to manage high concentration leachate from food waste and remove CODcr, TKN, TP, and salt effectively.

Genetic Characterization of Antigenic Variant Infectious Bursal Disease Virus (IBDV) in Chickens in Korea

  • Jong-Yeol Park;Ki-Woong Kim;Ke Shang;Sang-Won Kim;Yu-Ri Choi;Cheng-Dong Yu;Ji-Eun Son;Gyeong-Jun Kim;Won-Bin Jeon;In-Hwan Kim;Bai Wei;Min Kang;Hyung-Kwan Jang;Se-Yeoun Cha
    • Korean Journal of Poultry Science
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    • v.50 no.4
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    • pp.231-240
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    • 2023
  • Infectious bursal disease (IBD) is an acute, highly contagious, and immunosuppressive disease in young chickens, and causes considerable economic losses to the poultry industry. More than 30 years ago, an antigenic variant IBDV (avIBDV) was reported in chicken farms in the United States. Recently, a novel avIBDV exhibited clear differences in molecular characteristics compared with previous variant strains. This study investigated the molecular characteristics of recently isolated avIBDV strains in Korea. Strains of avIBDV were confirmed by reverse transcription PCR (RT-PCR) and were propagated in 10-day-old specific-pathogen-free (SPF) embryonated chicken eggs through chorioallantoic membrane (CAM) inoculation. Multiple sequence alignment and phylogenetic analyses of hypervariable regions VP2 gene revealed that the strains originated from two different avIBDV lineages (G2a and G2d). In our results, we confirmed the co-existence and prevalence of avIBDV genogroup G2a and G2d in chicken farms. It is necessary to study the protective efficacy of current vaccines against avIBDVs.

Real-time Nutrient Monitoring of Hydroponic Solutions Using an Ion-selective Electrode-based Embedded System (ISE 기반의 임베디드 시스템을 이용한 실시간 수경재배 양액 모니터링)

  • Han, Hee-Jo;Kim, Hak-Jin;Jung, Dae-Hyun;Cho, Woo-Jae;Cho, Yeong-Yeol;Lee, Gong-In
    • Journal of Bio-Environment Control
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    • v.29 no.2
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    • pp.141-152
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    • 2020
  • The rapid on-site measurement of hydroponic nutrients allows for the more efficient use of crop fertilizers. This paper reports on the development of an embedded on-site system consisting of multiple ion-selective electrodes (ISEs) for the real-time measurement of the concentrations of macronutrients in hydroponic solutions. The system included a combination of PVC ISEs for the detection of NO3, K, and Ca ions, a cobalt-electrode for the detection of H2PO4, a double-junction reference electrode, a solution container, and a sampling system consisting of pumps and valves. An Arduino Due board was used to collect data and to control the volume of the sample. Prior to the measurement of each sample, a two-point normalization method was employed to adjust the sensitivity followed by an offset to minimize potential drift that might occur during continuous measurement. The predictive capabilities of the NO3 and K ISEs based on PVC membranes were satisfactory, producing results that were in close agreement with the results of standard analyzers (R2 = 0.99). Though the Ca ISE fabricated with Ca ionophore II underestimated the Ca concentration by an average of 55%, the strong linear relationship (R2 > 0.84) makes it possible for the embedded system to be used in hydroponic NO3, K, and Ca sensing. The cobalt-rod-based phosphate electrodes exhibited a relatively high error of 24.7±9.26% in the phosphate concentration range of 45 to 155 mg/L compared to standard methods due to inconsistent signal readings between replicates, illustrating the need for further research on the signal conditioning of cobalt electrodes to improve their predictive ability in hydroponic P sensing.

A Study on the Water Reuse Systems (중수도개발연구(中水道開發研究))

  • Park, Chung Hyun;Lee, Seong Key;Chung, Jae Chul
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.4 no.4
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    • pp.113-125
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    • 1984
  • Water supply has been mainly dependent on the construction of the dams in Korea. It is difficult, however, to continue to construct dams for many reasons, such as the decrease of construction sites, the increase of construction costs, the compensation of residents in flooded areas, and the environmental effects. Water demands have increased and are expected to continue increasing due to the concentration of people in the cities, the rise of the living standard, and rapid industrial growth. It is acutely important to find countermeasures such as development of ground water, desalination, and recycling of waste water to cope with increasing water demands. Recycling waste water includes all means of supplying non-potable water for their respective usages with proper water quality which is not the same quality as potable water. The usages of the recycled water include toilet flushing, air conditioning, car washing, yard watering, road cleaning, park sprinkling, and fire fighting, etc. Raw water for recycling is obtained from drainage water from buildings, toilets, and cooling towers, treated waste water, polluted rivers, ground water, reinfall, etc. The water quantity must be considered as well as its quality in selecting raw water for the recycling. The types of recycling may be classified roughly into closed recycle systems and open recycle systems, which can be further subdivided into individual recycle systems, regional recycle systems and large scale recycle system. The treatment methods of wastewater combine biochemical and physiochemical methods. The former includes activated sludge treatment, bio-disc treatment, and contact aeration treatment, and the latter contains sedimentation, sand filtration, activated carbon adsorption, ozone treatment, chlorination, and membrane filter. The recycling patterns in other countries were investigated and the effects of the recycling were divided into direct and indirect effects. The problems of water reuse in recycle patterns were also studied. The problems include technological, sanitary, and operational problems as well as cost and legislative ones. The duties of installation and administrative organization, structural standards for reuse of water, maintenance and financial disposal were also studied.

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Amended Soil with Biopolymer Positively Affects the Growth of Camelina sativa L. Under Drought Stress (가뭄 조건 하에서 바이오폴리머 혼합 토양이 Camelina sativa L.의 생장에 미치는 긍정적 영향)

  • Lim, Hyun-Gyu;Kim, Hyun-Sung;Lee, Hyeon-Sook;Sin, Jung-Ho;Kim, Eun-Suk;Woo, Hyo-Seop;Ahn, Sung-Ju
    • Ecology and Resilient Infrastructure
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    • v.5 no.3
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    • pp.163-173
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    • 2018
  • The biopolymer (BP) used in this study is mainly composed of xanthan gum and ${\beta}$-glucan derived from microorganism and has been introduced as a novel material for soil stabilization. However, the broad applicability of BP has been suggested in the field of geotechnical engineering while little information is available about the effects of BP on the vegetation. The goal of this study is to find the BP effects on the growth of Camelina sativa L. (Camelina) under drought condition. For more thorough evaluation of BP effects on the plant growth, we examined not only morphological but also physiological traits and gene expression patterns. After 25 days of drought treatment from germination in the soil amended with 0, 0.25, 0.5, and 1% BP, we observed that the BP concentration was strongly correlated the growth of Camelina. When plants were grown under drought stress, Camelina in 0.5% BP mixture showed better physiological parameters of the leaf stomatal conductance, electrolyte leakage and relative water content compared to those in control soil without BP. Plant recovery rate after re-watering was higher and the development of lateral root was lower in BP amended soil. RNA expression of Camelina leaf treated with/without drought for 7 and 10 days showed that aquaporin genes transporting solutes at bio-membrane, CsPIP1;4, 2;1, 2;6 and TIP1;2, 2;1, were induced more in the plants with BP amendment and drought treatment. These results suggest that the soil amended with BP has a positive effect on the transport of nutrients and waters into Camelina by improving water retention in soil under drought condition.

Effect of n-3 Fatty Acid Deficiency on Fatty Acid Composition in Brain, Retina and Liver Using a Novel Artificial Rearing System (인공 사육 동물 모델 시스템을 이용한 n-3 지방산 결핍이 쥐의 뇌, 망막, 간의 지방산 조성에 미치는 영향)

  • Lim, Sun-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.4
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    • pp.466-475
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    • 2005
  • Docosahexaenoic acid (22:6n-3, DHA) is highly enriched in membrane of brain and retina, and plays an important role in maintaining an optimal function of the central nervous system. We investigated the effect of n-3 fatty acid deficiency on rat brain, retina and liver fatty acyl composition at two different ages (3 wks and 15 wks) under DHA deficient condition. Rat pups born to dams fed a diet with $3.1\%$ of total fatty acids as $\alpha-linolenic$ acid (LNA) were fed using an artificial rearing system either an n-3 deficient (n-3 Def) or n-3 adequate (n-3 Adq) diet. Both diets contained $17.1\%$ linoleic acid (LA) but the n-3 Adq diet also contained $3.1\%$ LNA. Rats consuming the n-3 Def diet showed a lower brain $(50\%\;in\;13\;wks\;and\;70\%\;in\;15\;wks,\;p<0.05)$ and retinal $(50\%\;in\;13\;wks\;and\;63\%\;in\;15\;wks,\;p<0.05)$ DHA than those on the n-3 Adq diet, which was largely compensated for by an increase in docosapentaenoic acid (22:5n-6, DPAn-6). In the liver of the n-3 Def group, the percentage of DHA decreased by $97\%$ at 3 wks of age with an apparent increase in DPAn-6 relative to the n-3 Adq group (p<0.05), while there was a $65\%$ lower liver DHA in n-3 Def group at 15 wks of age than the n-3 Adq group (p<0.05). Liver arachidonic acid (20:4n-6, AA) was increased at 3 wks of age but decreased at 15 wks of age in the n-3 Def group compared with n-3 Adq group (p<0.05). In conclusion, the replacement of DHA by DPAn-6 in brain and retina fatty acid composition may be related to the suboptimal function in spatial learning, memory and visual acuity. This artificial rearing method presents a first generation model for n-3 deficiency that is similar to the case of human nutrition that commonly employed two generation model.