• Title/Summary/Keyword: reaction control system

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Evaluation of horizontal gene transfer from genetically modified zoysiagrass to the indigenous microorganisms in isolated GMO field (GMO 격리포장에서의 유전자변형 들잔디로부터 토착미생물로의 수평유전자전달 평가)

  • Bae, Tae-Wung;Lee, Hyo-Yeon;Ryu, Ki-Hyun;Lee, Tae-Hyeong;Lim, Pyung-Ok;Yoon, Pill-Yong;Park, Sin-Young;Riu, Key-Zung;Song, Pill-Soon;Lee, Yong-Eok
    • Journal of Plant Biotechnology
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    • v.34 no.1
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    • pp.75-80
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    • 2007
  • The release of genetically modified organisms ($GMO_{s}$) into the environment has the potential risks regarding the possibility of gene transfer from $GMO_{s}$ to natural organisms and this needs to be evaluated. This study was conducted to monitor the possible horizontal gene transfer from herbicide-resistant zoysiagrass (Zoysia japonica Steud.) to indigenous microorganisms. We have first examined the effect of field-released GM zoysiagrass on the microbial flora in the gut of locust (Locusts mlgratoria). The microbial flora was analyzed through determining the 165 rDHA sequences of microorganisms. The comparison of the microbial flora in the gut of locusts that were captured at the field of GM zoysiagrass and of wild-type revealed that there is no noticeable difference between these two groups. This result indicates that the GM zoysiagrass does not have negative impact on microbial flora in the gut of locust. We then investigated whether the horizontal gene transfer occurred from GM zoysiagrass to microbes in soil, rhizosphere and faecal pellets from locusts by utilizing molecular tools such as Southern hybridization and polymerase chain reaction (PCR). When the total DNAs isolated from microbes in GM zoysiagrass and in wild-type zoysiagrass fields were hybridized with probes for bar or hpt gene, no hybridization signal was detected from both field isolates, while the probes were hybridized with DNA from the positive control. Absence of these genes in the FNAs of soil microorganisms as well as microbes in the gut of locust was further confirmed by PCR. Taken together, our data showed that horizontal gene transfer did not occur in this system. These results further indicate that frequencies of transfer of engineered plant DNA to bacteria are likely to be negligible.

Lack of Association of the NPAS2 Gene Ala394Thr Polymorphism (rs2305160:G>A) with Risk of Chronic Lymphocytic Leukemia

  • Rana, Sobia;Shahid, Adeela;Ullah, Hafeez;Mahmood, Saqib
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.17
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    • pp.7169-7174
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    • 2014
  • Background: NPAS2 is a product of the circadian clock gene. It acts as a putative tumor suppressor by playing an important role in DNA damage responses, cell cycle control and apoptosis. Chronic lymphocytic leukemia (CLL) appears to be an apoptosis related disorder and alteration in the NPAS2 gene might therefore be directly involved in the etiology of CLL. Here, the Ala394Thr polymorphism (rs2305160:G>A) in the NPAS2 gene was genotyped and melatonin concentrations were measured in a total of seventy-four individuals, including thirty-seven CLL cases and an equal number of age- and sex-matched healthy controls in order to examine the effect of NPAS2 polymorphism and melatonin concentrations on CLL risk in a Pakistani population. Materials and Methods: Genotyping of rs2305160:G>A polymorphism at NPAS2 locus was carried out by amplification refractory mutation system-polymerase chain reaction (ARMS-PCR). Melatonin concentrations were determined by enzyme linked immunosorbent assay (ELISA). Statistical analysis was performed using Statistical Package for Social Sciences software. Results: Our results demonstrated no association of the variant Thr genotypes (Ala/Thr and Thr/Thr) with risk of CLL. Similarly, no association of rs2305160 with CLL was observed in either females or males after stratification of study population on a gender basis. Moreover, when the subjects with CLL were further stratified into shift-workers and non-shift-workers, no association of rs2305160 with CLL was seen in either case. However, significantly low serum melatonin levels were observed in CLL patients as compared to healthy subjects (p<0.05). Also, lower melatonin levels were seen in shift-workers as compared to non-shift-workers (p<0.05). There was no significant difference (p>0.05) in the melatonin levels across NPAS2 genotypes in all subjects, subjects with CLL who were either shift workers or non-shift-workers. General Linear Model (GLM) univariate analysis revealed no significant association (p>0.05) of the rs2305160 polymorphism of the NPAS2 gene with melatonin levels in any of the groups. Conclusions: While low melatonin levels and shift-work can be considered as one of the risk factors for CLL, the NPAS2 rs2305160 polymorphism does not appear to have any association with risk of CLL in our Pakistani population.

Quantitative Analysis of Bacillus amyloliquefaciens GR4-5 in Soil (Bacillus amyloliquefaciens GR4-5 균주의 토양 내 정량 분석)

  • Kim, Dayeon;Kim, Byung-Yong;Ahn, Jae-Hyung;Weon, Hang-Yeon;Kim, Sung-Il;Kim, Wan-Gyu;Song, Jaekyeong
    • Korean Journal of Organic Agriculture
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    • v.23 no.4
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    • pp.847-858
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    • 2015
  • Bacillus amyloliquefaciens GR4-5 was isolated from the rhizosphere soil of Korean ginseng and displayed broad-spectrum suppression of ginseng root rot pathogens. The survivability of B. amyloliquefaciens GR4-5 in soil was investigated under three different conditions; indoor, outdoor - of which soil was put in 14 mL tube after treatment - and field environments. Soil samples were collected over a four-week period from three experimental designs, and assessed for 16S rRNA gene copy number by quantitative polymerase chain reaction (qPCR). In outdoor condition, the 16S rRNA gene copy number of Bacillus spp. was 8.35 log copies g $soil^{-1}$ immediately after the GR4-5 treatment. Two weeks later, the 16S rRNA gene copy number of Bacillus spp. (6.70 log copies g $soil^{-1}$) was similar to that of the control (6.38 log copies g $soil^{-1}$). In indoor condition, the 16S rRNA gene copy number of Bacillus spp. maintained in a certain level for a longer period than those in outdoor and field. The 16S rRNA gene copy number of Bacillus spp. in field experiment was reduced faster than that of outdoor condition. Our results show that B. amyloliquefaciens GR4-5 can survive in bulk soil for 1 week, indicating its potential use as a biocontrol agent following 7 day application intervals. This study presents that outdoor microcosm system design could be a useful method to assess easily the survivability of beneficial microorganisms.

Effects of Visual Information Blockage on Landing Strategy during Drop Landing (시각 정보의 차단이 드롭랜딩 시 착지 전략에 미치는 영향)

  • Koh, Young-Chul;Cho, Joon-Haeng;Moon, Gon-Sung;Lee, Hae-Dong;Lee, Sung-Cheol
    • Korean Journal of Applied Biomechanics
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    • v.21 no.1
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    • pp.31-38
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    • 2011
  • This study aimed to determine the effects of the blockage of visual feedback on joint dynamics of the lower extremity. Fifteen healthy male subjects(age: $24.1{\pm}2.3\;yr$, height: $178.7{\pm}5.2\;cm$, weight: $73.6{\pm}6.6\;kg$) participated in this study. Each subject performed single-legged landing from a 45 cm-platform with the eyes open or closed. During the landing performance, three-dimensional kinematics of the lower extremity and ground reaction force(GRF) were recorded using a 8 infrared camera motion analysis system (Vicon MX-F20, Oxford Metric Ltd, Oxford, UK) with a force platform(ORG-6, AMTI, Watertown, MA). The results showed that at 50 ms prior to foot contact and at the time of foot contact, ankle plantar-flexion angle was smaller(p<.05) but the knee joint valgus and the hip flexion angles were greater with the eyes closed as compared to with the eyes open(p<.05). An increase in anterior GRF was observed during single-legged landing with the eyes closed as compared to with the eyes open(p<.05). Time to peak GRF in the medial, vertical and posterior directions occurred significantly earlier when the eyes were closed as compared to when the eyes were open(p<.05). Landing with the eyes closed resulted in a higher peak vertical loading rate(p<.05). In addition, the shock-absorbing power decreased at the ankle joint(p<.05) but increased at the hip joints when landing with the eyes closed(p<.05). When the eyes were closed, landing could be characterized by a less plantarflexed ankle joint and more flexed hip joint, with a faster time to peak GRF. These results imply that subjects are able to adapt the control of landing to different feedback conditions. Therefore, we suggest that training programs be introduced to reduce these injury risk factors.

Protective Effect of Soybean Sauce and Melanoidin on Lipid Oxidation in Rats Fed High PUFA Oils (고도불포화지방산 함량이 높은 유지를 섭취시킨 흰쥐에서 양조간장과 멜라노이딘의 지질산화 억제효과)

  • 이상조;류승희;이영순;송영선;문갑순
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.6
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    • pp.913-920
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    • 2003
  • Soybean sauce fermented with soybean and wheat, has been a major condiment of Korean diets from centuries ago. Melanoidin, a brown pigment generally found in various food systems, is a final product produced in amino-carbonyl reaction during soybean sauce processing. Antioxidative activities of soybean sauce and melanoidin were investigated in vitro system using linoleic acid emulsion. Soybean sauce and glucose-lysine model melanoidin showed the stronger antioxidative effect than control by ferric thiocyanate and conjugated diene assays. In addition, DPPH radical scavenging effect of soybean sauce was higher than melanoidin, which was ascribed to soluble peptide and low molecular protein existing in soybean sauce. To ascertain antioxidative effect of dietary soybean sauce and melanoidin in vivo, the male Wister rats were fed 10% soybean sauce or 10% glucose-lysine model melanoidin with corn oil or fish oil for 5 weeks. Fatty acid compositions in liver and plasma were influenced by oil source. Therefore, EPA and DHA contents of fish oil group were higher than those of corn oil group. When the inhibitory effect of soybean sauce and melanoidin on lipid peroxidation using TBARS methods was measured, fish oil group (FC) showed higher malondialdehyde (MDA) content than corn oil group (CC). However, supplementation of soybean sauce and melanoidin to fish oil group attenuated MDA formation. In the levels of phosphatidyl choline hydroperoxide (PCOOH) in liver and plasma by CL (chemiluminescence)-HPLC method, PCOOH in FC group was significantly higher than that of CC group both in liver and plasma. Supplementation of soybean sauce to fish oil groups significantly inhibited the formation of PCOOH in plasma and liver, while melanoidin suppressed hepatic PCOOH formation. Based on these results, it can be suggested that soybean sauce possesses stronger antioxidative potential than melanoidin.

Pathogenesis and pathogenicity for the porcine circovirus 2 Korean isolates in weaned pigs (국내 분리 porcine circovirus 2의 이유자돈에 대한 병원성 시험연구)

  • Roh, In-Soon;Lee, Kyung-Woo;Kim, Jae-Hoon;Han, Jeong-Hee;Jean, Young-Hwa
    • Korean Journal of Veterinary Research
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    • v.47 no.2
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    • pp.175-185
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    • 2007
  • This study was carried out to investigate the pathogenesis and pathogenicity of the porcine circovirus type 2 (PCV2) Korean isolate from weaned pigs. Twenty four weaned pigs, PCV2, porcine reproductive and respiratory syndrome virus (PRRSV) and porcine parvovirus (PPV) antibodies free, were allocated to 4 groups (n = 6). Six pigs were inoculated intranasally with PCV2 alone, 6 with PCV2 and PRRSV, 6 with the combined PCV2/PRRSV/PPV inoculum, and 6 were remained as a uninoculated negative control. Pigs were killed 3 and 6 weeks after inoculation and tissue samples examined for gross and microscopic lesions and for the presence of PCV2 antigens and nucleic acids. Experimentally inoculated pigs were evaluated for 3 considerations: 1. development of postweaning multisystemic wasting syndrome (PMWS), 2. distribution of viral antigens by immunohistochemistry and polymerase chain reaction (PCR), and 3. cytokine mRNA levels in lymph nodes. Pigs inoculated with PCV2/PRRSV/PPV showed typical clinical signs, gross findings, and histopathologic characteristics of PMWS. In the PCV2/PRRSV/PPV inoculated group, the PCV2 antigen was widely distributed in various parenchymal organs such as brain, spinal cord, tonsil, lymph nodes, lung, heart, liver, kidney, spleen, and peyer's patch. Lymph node mRNA expression of IL-$1{\alpha}$, IL-2R and IL-8 was determined by real-time PCR. The pigs of PCV2/PRRSV and PCV2/PRRSV/PPV inoculation group, the mRNA expression was characterized by a decrease of IL-$1{\alpha}$, IL-2R and IL-8. The decrease of cytokine mRNA represent the state of T cell immuno-suppression in pig, and nicely support the evidence for the impairment of immune system in pigs with PMWS. In conclusion, PCV2 infection and some additional infectious causes such as PRRSV and/or PPV are warranted for the presence of PMWS in weaned pigs in Korea.

The Physicochemical Characteristics of Marinated Beef Galbi under Different Cooking Conditions (양념 소갈비의 조리과정에서의 물리화학적 특성 평가)

  • Hong, Sang-Pil;Kim, Young-Ho;Lee, Nam-Hyouck;Heo, Yeong-Uk
    • Journal of the Korean Society of Food Culture
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    • v.28 no.1
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    • pp.78-88
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    • 2013
  • Marinated beef galbi is a traditional Korean dish cooked with soy sauce, pear juice, onion, sesame oil, and sugar. However, there are many differences in beef galbi, including flavor and physicochemical aspects, depending on cooking conditions. Therefore, the physicochemical characteristics of marinated beef galbi prepared through various recipes was evaluated for its effects on pH, texture, aging, proteolysis, heating conditions, cooking time, and flavor compounds (pyrazines, IMPs, or FAAs). There were significant differences in salt concentration (0.8~3.03%), pH (4.89~6.22), and solid soluble contents (1.34-6.31 Brix) between recipes in this study. In the Pearson assay for sensory evaluation, overall preference correlated well with texture (a well-known sensory attribute in meat evaluation). Controlling the pH of meat through soaking in lemon solution, alkali water, phosphate, and baking powder solution, improved water holding capacity as much as 9 to 15% compared with the control. The myofibril index (MFI) of marinated meat stored at $4^{\circ}C$ increased 32% with 24 hours of aging and reached 39% at 48 hours of aging, and its fragmentation was observed through microscopy. SDS-PAGE showed hydrolysis of acid-soluble collagen by the pear juice, possibly related to meat tenderness. On the basis of surface temperature, the cooking time was estimated to be 8 minutes with pan heating at $170^{\circ}C$, 6 minutes at $270{\sim}300^{\circ}C$, and 4 minutes with charcoal at $700{\sim}900^{\circ}C$. Different pyrazine compounds, such as 2-methyl-3-phenylpyrrol(2,3-b) pyrazine (the typical product of the browning reaction) was mainly detected, and IMP (one of the main taste compounds in beef) was in higher amounts with the charcoal treatment, potentially related to its flavor preference among treatments. Our results demonstrate an effective case study and cooking system for beef galbi.

Development of Hepatitis C Virus (HCV) Genome-Targeting Hammerhead Ribozyme Which Activity Can Be Allosterically Regulated by HCV NS5B RNA Replicase (C형 간염바이러스(HCV)의 NS5B RNA Replicase에 의해 그 활성이 조절되는 HCV지놈 표적 Hammerhead 리보자임 개발)

  • Lee, Chang-Ho;Lee, Seong-Wook
    • Korean Journal of Microbiology
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    • v.43 no.3
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    • pp.159-165
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    • 2007
  • For the development of basic genetic materials for specific and effective therapeutic approach to suppress multiplication of hepatitis C virus (HCV), HCV internal ribosome entry site (IRES)-targeting hammerhead ribozyme which activity is allosterically regulated by HCV regulatory protein, NS5B RNA replicase, was developed. The ribozyme targeted most effectively to +382 nucleotide (nt) site of HCV IRES RNA. The allosteric ribozyme was designed to be composed of sequence of RNA aptamer to HCV NS5B, communication module sequence which can transfer structural transition for inducing ribozyme activity upon binding NS5B to the aptamer, and sequence of ribozyme targeting +382 nt of HCV IRES. Noticeably, we employed in vitro selection technology to identify the most appropriate communication module sequence which can induce ribozyme activity depending on the US5B protein. We demonstrated that the ribozyme was nonfunctional either in the absence of any proteins or in the presence of control bovine serum albumin. In sharp contrast, the allosteric ribozyme can induce activity of cleavage reaction with HCV IRES RNA in the presence of the HCV NS5B protein. This allosteric ribozyme can be used as lead compound for specific and effective anti-HCV agent, tool for highthroughput screening to isolate lead chemicals for HCV therapeutics, and ligand for biosensor system for HCV diagnosis.

Transformation of the Diatom Phaeodactylum tricornutum with its Endogenous (E)-4-Hydroxy-3-methylbut-2-enyl Diphosphate Reductase Gene (Phaeodactylum tricornutum의 (E)-4-Hydroxy-3-methylbut-2-enyl Diphosphate Reductase 유전자의 형질전환)

  • Shin, Bok-Kyu;Jung, Yu-Jin;Kim, Sang-Min;Pan, Cheol-Ho
    • Journal of Applied Biological Chemistry
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    • v.58 no.3
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    • pp.273-279
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    • 2015
  • Phaeodactylum tricornutum is a model diatom that its genomic information and biological tools are well established. In this study, a gene encoding (E)-4-hydroxy-3-methylbut-2-enyl diphosphate reductase (PtHDR), a terminal enzyme of the methylerythritol phosphate pathway regulating chlorophyll and carotenoid biosynthesis, was isolated from P. tricornutum. The isolated gene was cloned into pPha-T1 vector containing fcpA promoter to prepare pPha-T1-HDR plasmid. As a positive control, pPha-T1-eGFP plasmid was constructed with egfp gene. Stable nuclear transformation was carried out with these plasmids by particle bombardment method and zeocin resistant colonies of P. tricornutum were selected on f/2 agar plate. In result, transformation efficiency was evaluated according to the amount of plasmid DNA coated with gold particles. Integration of introduced plasmids was confirmed with genomic DNA of each transformant by polymerase chain reaction. The eGFP fluorescence was visible in the cytoplasm, indicating that eGFP was successively expressed in P. tricornutum system. The transcript level of exogenous Pthdr gene was evaluated with the obtained transformants. The results presented here demonstrated that introduction of Pthdr gene into P. tricornutum chromosome succeeded and expression of PtHDR was enhanced under the fcpA promoter.

Effects of Arabinoxylan Rice Bran and Exercise Training on Immune Function and Inflammation Response in Lipopolysaccharide-stimulated Rats (유색미 겨 아라비녹실레인과 운동트레이닝이 Lipopolysaccharide 처치된 흰쥐의 면역인자 및 염증억제에 미치는 영향)

  • Son, Hee-Jeong;Kim, Hyung-Jun;Chae, Jeong-Hoon;Kwon, Hyung-Tae;Yeo, Hyo-Seong;Eo, Su-Ju;Leem, Yea-Hyun;Kim, Hyo-Jeong;Kim, Chang-Keun
    • Journal of Applied Biological Chemistry
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    • v.55 no.1
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    • pp.41-46
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    • 2012
  • Arabinoxylan (Ara) rice bran has been shown to be a potent biological response modifier as manifested by stimulation of different arms of the immune system. We examined the effects of Ara rice bran and exercise on the immune function and cytokines in lipopolysaccharide (LPS)-stimulated rats. As the results, tumor necrosis factor-${\alpha}$ as representative inflammatory cytokines showed a significantly lower in Ara supplement group, thus the Ara rice bran had a higher inhibitory activity than the both exercise and control group. However, 4 weeks of exercise training significantly increased inflammatory reactions rather than treatment with Ara in LPS-treated rats. The Ara rice bran acted to decrease the inflammatory reaction. These results suggest that the supplement of Ara rice bran is likely contribute to inflammation response and the Ara rice bran can be used as a possible safe alternative to the immunotherapeutic modalities.