• Title/Summary/Keyword: Biological fixation

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Expression of EuNOD-ARP1 Encoding Auxin-repressed Protein Homolog Is Upregulated by Auxin and Localized to the Fixation Zone in Root Nodules of Elaeagnus umbellata

  • Kim, Ho Bang;Lee, Hyoungseok;Oh, Chang Jae;Lee, Nam Houn;An, Chung Sun
    • Molecules and Cells
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    • v.23 no.1
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    • pp.115-121
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    • 2007
  • Root nodule formation is controlled by plant hormones such as auxin. Auxin-repressed protein (ARP) genes have been identified in various plant species but their functions are not clear. We have isolated a full-length cDNA clone (EuNOD-ARP1) showing high sequence homology to previously identified ARP genes from root nodules of Elaeagnus umbellata. Genomic Southern hybridization showed that there are at least four ARP-related genes in the genome of E. umbellata. The cDNA clone encodes a polypeptide of 120 amino acid residues with no signal peptide or organelle-targeting signals, indicating that it is a cytosolic protein. Its cytosolic location was confirmed using Arabidopsis protoplasts expressing a EuNOD-ARP1:smGFP fusion protein. Northern hybridization showed that EuNOD-ARP1 expression was higher in root nodules than in leaves or uninoculated roots. Unlike the ARP genes of strawberry and black locust, which are negatively regulated by exogenous auxin, EuNOD-ARP1 expression is induced by auxin in leaf tissue of E. umbellata. In situ hybridization revealed that EuNOD-ARP1 is mainly expressed in the fixation zone of root nodules.

Lipid N-formylation Occurs During Fixation with Formalin

  • Kim, Min Jung;Lim, Heejin;Kim, Muwoong;Choi Yuri;Nguyen, Thy N.C.;Park, Seung Cheol;Kim, Kwang Pyo;Jung, Junyang;Kim, Min-Sik
    • Mass Spectrometry Letters
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    • v.13 no.2
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    • pp.35-40
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    • 2022
  • Human tissues and organs can be preserved intact by fixation with formalin for the future analysis of biomolecules of interest. With the advances in high-throughput methods, numerous protocols have been developed and optimized to attain the most pathophysiological information out of biomolecules, including RNA and proteins, in formalin-fixed samples. However, there is no systematic study to examine the effects of formalin fixation on the lipidome of biological samples in a global fashion. In this study, we conducted a mass spectrometry-based analysis to survey the alteration in the lipidome of mice brains by fixation methods. A total of 308 lipids were quantitatively measured using triple quadrupole mass spectrometry. We found that most were unchanged after formalin fixation except for a few lipid classes such as phosphatidylethanolamine.

Ultrastructures of Ptilota filicina (Rhodophyta) by High Pressure Freezing(HPF): Comparison of HPF Fixation and Chemical Fixation (High Pressure Freezing (HPF)을 이용한 조류 Ptilota filicina의 미세구조 관찰:HPF 고정법과 화학 고정법의 비교)

  • Lee, Sang-Hee;Kim, Youn-Joong;Jeong, Jong-Man;Kim, Jin-Gyu;Kim, Young-Min;Kweon, Hee-Seok;Moon, Won-Jin;Lee, Seok-Hoon
    • ALGAE
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    • v.21 no.4
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    • pp.479-483
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    • 2006
  • In preparation of the biological samples for electron microscopy, the chemical fixation by glutaraldehyde, paraformaldehyde, and OsO4 has been generally used for a long time. However, the chemical fixation method has some problems: the infiltration time is a little bit long and the ultrastructure of cell or tissue transforms before complete fixation of sample. So, recently, cryo-fixation is considered more often in biomedical field. In this study, we compared High Pressure Freezing (HPF) method with chemical fixation method using a algal sample (Ptilota filicina J. Agardh), which was difficult to fix using chemical fixation method. In chloroplast, the ultrastructure of thylakoid lamella and phycobilisome can not show clearly by chemical fixation. In this study we could observe the ultrastructure of thylakoid lamella and phycobilisome of chloroplast very clearly using HPF fixation. An improved images of ultrastructures of nucleus, mitochondrion and floridean starch could obtain. These results suggest that HPF method is very useful method in algal specimen for electron microscopy.

EFFECTS OF THE HERBICIDE, BUTACHLOR, ON NITROGEN FIXATION IN PHOTOTROPHIC NONSULFUR BACTERIA

  • Lee, Kyung-Mi;Kim, Jai-Soo;Lee, Hyun-Soon
    • Environmental Engineering Research
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    • v.12 no.4
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    • pp.136-147
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    • 2007
  • In an effort to identify possible microbes for seeking bioagents for remediation of herbicide-contaminated soils, seven species of phototrophic nonsulfur bacteria (Rhodobacter capsulatus and sphaeroides, Rhodospirillum rubrum, Rhodopseudomonas acidophila, blastica and viridis, Rhodomicrobium vannielii) were grown in the presence of the herbicide, butachlor, and bacterial growth rates and nitrogen fixation were measured with different carbon sources. Under general conditions, all species showed 17-53% reductions in growth rate following butachlor treatment. Under nitrogen-fixing conditions, Rb. capsulatus and Rs. rubrum showed 1-4% increases in the growth rates and 2-10% increases in nitrogen-fixing abilities, while the other 5 species showed decreases of 17-47% and 17-85%, respectively. The finding that Rp. acidophila, Rp. blastica, Rp. viridis and Rm. vannielii showed stronger inhibitions of nitrogenase activity seems to indicate that species in genera Rhodobacter and Rhodospirillum are less influenced by butachlor than those in Rhodopseudomonas and Rhodomicrobium in terms of nitrogen-fixing ability. Overall, nitrogenase activity was closely correlated with both growth rate and glutamine synthetase activity (representing nitrogen metabolism). When the carbon sources were compared, pyruvate (three carbons) was best for all species in terms of growth rate and nitrogen fixation, with malate (four carbons) showing intermediate values and ribose(five carbons) showing the lowest; these trends did not change in response to butachlor treatment. We verified that each of the 7 species had a plasmid ($12.2{\sim}23.5\;Kb$). We found that all 7 species could use butachlor as a sole carbon source and 3 species were controlled by plasmid-born genes, but it is doubtful whether plasmid-born genes were responsible to nitrogen fixation.

Biological Fixation of Carbon Dioxide by Synechocystis PCC 6803 (Synechocystis PCC 6803에 의한 이산화탄소의 생물학적 고정화)

  • 김장규;원성호;김남기
    • KSBB Journal
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    • v.13 no.1
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    • pp.101-107
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    • 1998
  • Carbon dioxide is estimated to be responsible for 60% of the global warming effect, and this percentage is tending upward. Studies on removal and fixation of $CO_2$ in the flue gas are recognized as one of the important roles of the future biotechnology. Photobiological systems have considerably higher photosynthetic efficiency than conventional biomass system. The experiment for the photosynthetic fixation of $CO_2$ and the biomass production was performed with various initial cell concentration in a tubular photobioreactor and a bubble column $CO_2$ contactor with a gas sparger of $CO_2$ -enriched air(0.03~20%). Synechocystis PCC 6803 could grow at 10~20% $CO_2$ content under pH control. The highest specific growth rate, 0.0258 $h^{-1}$ , was obtained at 5% $CO_2$-air mixture. The maximum cell production rate, 0.2784 g/L.day, was obtained when the initial cell concentration was 0.45 g/L at 5% $CO_2$ -air mixture. The maximum cell concentration was 2.03 g/L in the tubular photobioreactor when the light intensity was $45.5{\mu}$ $E/m^2$ . s. This system showed 0.482 g $CO_2$ /L . day of the $CO_2$ fixation.

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Improvement of Histopathological Sample Preparation by Employing Microwave Heating Method on Frozen Section Specimens

  • Ahn, Seung-Ju
    • Biomedical Science Letters
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    • v.13 no.4
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    • pp.361-368
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    • 2007
  • Biological samples can be fixed either by chemical method by using chemical solution or physical methods by using heat treatment. The problem in traditional heat fixation is unsatisfactory quality due to uneven heat conduction in specimen and loss of inner cell contents. Chemical fixation method also bears several intrinsic problems like the limit in specimen size, time consumption in fixative impregnation, and loss of low molecular weight cell components. These factors deteriorate the quality of fixed specimen, thus limit the magnification and contrast of tissue pictures. Microwave heat has been reported to be a good alternative to current chemical methods to overcome these problem. In this study, we tried to introduce the microwave energy method to routine fixation work in hospital. We replaced chemical fixative with saline to provide moderate reaction condition, and used frozen section to reduce time for sample preparation. Temperature was measured at each experiment. The fixation of rat kidney tissue with 2.45 GHz electromagnetic wave and saline showed similar result to the control group fixed with traditional chemical method. Human tumor tissue fixed with 2.45 GHz electromagnetic in frozen section was improved in terms of histochemistry of PAS and immunohistochemistry of tumor marker like cytokeratin. Total turnaround time was reduced from $24\sim38$ h to to $2\sim4$ h. In conclusion, the quality of samples prepared by microwave heating method was at least as good as that of traditional method. If the condition for the fixation of different specimens is standardized, this new method could be applied to routine work in hospital, and could save working time as well.

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Microwave fixation of Setaria Digitata for scanning electron microscopy (선충류의 주사전자현미경적 관찰을 위한 마이크로웨이브 고정법)

  • Lee, San-Soo;Cho, Kyoung-Oh;Shin, Kil-Sang;Shin, Sung-Shik
    • Korean Journal of Veterinary Research
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    • v.47 no.2
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    • pp.203-207
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    • 2007
  • Conventional processing of biological materials including nematode parasites for scanning electron microscopy includes fixation with glutaraldehyde and osmium, followed by dehydration in an ascending grade of ethanol, and finally freeze drying. This procedure takes about 8 to 12 h depending on the characteristics of samples. Microwave irradiation of 2,450 MHz enhance the action of cross-linking fixatives and can greatly accelerate various stages of tissue processing. In this study, samples of nematode parasites, Setaria digitata, were fixed by a combination of conventional chemical fixation and the microwave irradiation during the process. The microwave irradiation was also incorporated in the serial dehydration process with ethanol. The complete procedure from the initial fixation to the completion of dehydration with ethanol was reduced to 1 h with good preservation of the ultrastructural details of the specimens.

Effects of legume mixture on nitrogen fixation and transfer to grasses in spring paddy field

  • Lee, H.
    • Korean Journal of Organic Agriculture
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    • v.19 no.spc
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    • pp.161-164
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    • 2011
  • Nitrogen fixation by legumes can be valuable sources for organic farming. This study was to investigate the effect of different legume mixtures on nitrogen fixation and transfer to grasses on spring paddy field. Three different mixtures were used (rye+hairy vetch, Italian ryegrass+crimson clover, oat+pea) in a randomized complete block design with three replications and sowed in pots with different sowing rate (5:5 rye:hairy vetch,7:3=Italian:crimson, 6:4=oat:pea) on early March. $(^{15}NH_4)SO_4$ solution at. 99.8 atom%$^{15}N$ was applied to the each pot at the rate of 2kg N $ha^{-1}$ on $16^{th}$ April. Forage were harvested at ground level in heading stage and separated into legume and grass. Total N content and $^{15}N$ value were determined using a continuous flow stable isotope ratio mass spectrometry. DM yield of rye+vetch, Italian+crimson and oat+pea were 6,607, 3,213 and 4,312kg/ha, respectively. Proportion of N from fixation was 0.73(rye+vetch), 0.42(Italian+crimson) and 0.93(oat+pea). The percentages of N transfer from legume to grass were from 61% to 24% in different method by treatments and -35% to 21% in isotope dilution method.

The Influence of Fixation Rigidity on Intervertebral Joints - An Experimental Comparison between a Rigid and a Flexible System

  • Kim, Won-Joong;Lee, Sang-Ho;Shin, Song-Woo;Rivard, Charles H.;Coillard, Christine;Rhalmi, Souad
    • Journal of Korean Neurosurgical Society
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    • v.37 no.5
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    • pp.364-369
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    • 2005
  • Objective: Spinal instrumentation without fusion often fails due to biological failure of intervertebral joints (spontaneous fusion, degeneration, etc). The purpose of this study is to investigate the influence of fixation rigidity on viability of intervertebral joints. Methods: Twenty pigs in growing period were subjected to posterior segmental fixation. Twelve were fixed with a rigid fixation system(RF) while eight were fixed with a flexible unconstrained implant(FF). At the time of the surgery, a scoliosis was created to monitor fixation adequacy. The pigs were subjected to periodic radiological examinations and 12pigs (six in RF, six in FF) were euthanized at 12-18months postoperatively for analysis. Results: The initial scoliotic curve was reduced from $31{\pm}5^{\circ}$ to $27{\pm}8^{\circ}$ in RF group (p=0.37) and from $19{\pm}4^{\circ}$ to $17{\pm}5^{\circ}$ in FF group (p=0.21). Although severe disc degeneration and spontaneous fusion of facet joints were observed in RF group, disc heights of FF group were well maintained without major signs of degeneration. Conclusion: The viability of the intervertebral joints depends on motion spinal fixation. Systems allowing intervertebral micromotion may preserve the viability of intervertebral discs and the facet joint articular cartilages while maintaining a reasonably stable fixation.

Nitrogen Balance and Biological Nitrogen Fixation of Soybean in Soybean-Barley Cropping System

  • Park Sei Joon;Kim Wook Han;Lee Jae Eun;Kwon Young Up;Shin Jin Chul;Ryu Yong Hwan;Seong Rak Chun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.50 no.1
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    • pp.1-4
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    • 2005
  • This experiment was conducted to investigate the soil nitrogen credit of biological nitrogen fixation (BNF) and the nitrogen balance of soybean in soybean-barley cropping systems. Soybean cultivar, Shinpaldalkong2 and barley cultivar, Olbori, were used in soybean mono-cropping (SM), barley monocropping (BM), and barley­soybean double cropping system. The barley-soybean double cropping system was treated with two different levels of nitrogen fertilizers, 0 nitrogen fertilizer (BS-F0), and standard nitrogen fertilizer (BS-F1). Nitrogen and organic matter concentrations in soil of BS-F1 plot on October, 2001 were increased $4.8\%\;and\;5.9\%$, respectively, compared with those on October, 2000. The ranges of BNF rate in soybean were $69.1\~ 88.2\%$ in two years, and the rate was the highest in BS-F0 plot and the lowest in SM plot. The ranges of nitrogen harvest index (NHI) in all treatments were $83.9\~86.7\%$. The yield was 270 kg/10a in BS­F1 plot and 215 kg/10a in BS-F0 plot. However, the nitrogen balances were +0.6 kg/10a of gain of soil nitrogen in BS-F0 plot and -0.4 kg/10a of loss of soil nitrogen in BS-F1 plot. In comparisons of SM and BS-F1 plots, although the seed yields were similar in two plots, the loss of soil nitrogen was higher in SM than BS-F1 plot. Overall, our results suggest that barley-soybean double cropping system was more effective in respect to seed productivity and soil nitrogen conservation than soybean monocropping system, and the N credit to following crops by soybean cultivation was identified in soybean double cropping system.