• Title/Summary/Keyword: Single Cell

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Single Cell Oil-Recent Trends in Microbial Production and Utilization (미생물에 의한 단세포유지의 생산과 이용)

  • Kim, Yong-Ro;Yoon, Suk Hoo
    • Korean Journal of Food Science and Technology
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    • v.47 no.6
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    • pp.687-697
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    • 2015
  • With the shortage of edible fats and oils and depletion of fossil fuels in many countries, microbial lipids is emerging as one of the most promising sources of fats and oils in the global market. Oleaginous microorganisms, also called single cell oils (SCOs), can accumulate lipids more than 25% in the cell volume. Triacylglycerols are the major storage lipids. SCOs offer several advantages for lipid production as follows: SCOs have short life span which would shorten production time, cultivation conditions are not affected by climate and place; the production process is easy to control. There are a number of oleaginous yeasts, molds, and microalgae. Furthermore, the lipid productivity of SCOs can be enhanced through strain improvement and the optimization of cultivation conditions. The new strains developed using recent advanced biotechnical methods showed greatly improved oleaginicity. Further, hydrolysates of lignocellulosic materials can be used as carbon sources for economic production of SCO.

Effects of irradiation on the mRNA expression of the osteocalcin and osteopontin in MC3T3-E1 osteoblastic cell line (MC3T3-E1 조골세포주의 osteocalcin과 osteopontin mRNA 발현에 미치는 방사선의 영향)

  • Cho Su-Beom;Lee Sang-Rae;Koh Kwang-Joon
    • Imaging Science in Dentistry
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    • v.33 no.3
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    • pp.179-185
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    • 2003
  • Purpose: To investigate the effects of irradiation on the phenotypic expression of the MC3T3-El osteoblastic cell line, particularly on the expression of osteocalcin and osteopontin. Materials and Methods: Cells were irradiated with a single dose of 0.5, 1,4, and 8 Gy at a dose rate of 5.38 Gy/min using a cesium 137 irradiator. After the specimens were harvested, RNA was extracted on the 3rd, 7th, 14th, and 21st day after irradiation. The RNA strands were reverse-transcribed and the resulting cDNAs were subjected to amplification by PCR. Results: The irradiated cells demonstrated a dose-dependent increase in osteocalcin and a dose-dependent decrease in osteopontin mRNA expression compared with the non-irradiated control group, The amount of osteocalcin mRNA expression decreased significantly at the 3rd day after irradiation of 0,5, 1,4, and 8 Gy, and also decreased significantly at the 3rd, 14th, and 21 st day after irradiation in the 8 Gy exposed group compared with the control group, The degree of osteopontin mRNA expression increased significantly at the 7th day after irradiation of 0,5, 1,4, and 8Gy, Conclusion: These results showed that each single dose of 0,5, 1, 4, and 8 Gy influenced the mRNA expression of osteocalcin and osteopontin associated with the calcification stage of osteoblastic cells, suggesting that each single dose affected bone formation at the cell level.

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Development of a real-time PCR method for detection and quantification of the parasitic protozoan Perkinsus olseni

  • Gajamange, Dinesh;Yoon, Jong-Man;Park, Kyung-Il
    • The Korean Journal of Malacology
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    • v.27 no.4
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    • pp.387-393
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    • 2011
  • The objective of this study was to develop a real-time PCR method for the rapid detection and quantification of the protozoan pathogen Perkinsus olseni using a TaqMan probe. For the standard, genomic DNA was extracted from $10^5$ in vitro-cultured P. olseni trophozoites, and then 10-fold serial dilutions to the level of a single cell were prepared. To test the reliability of the technique, triplicates of genomic DNA were extracted from $5{\times}10^4$ cells and 10-fold serial dilutions to the level of 5 cells were prepared. The standards and samples were analyzed in duplicate using an $Exicycler^{TM}$ 96 real-time quantitative thermal block. For quantification, the threshold cycle ($C_T$) values of samples were compared with those obtained from standard dilutions. There was a strong linear relationship between the $C_T$ value and the log concentration of cells in the standard ($r^2$ = 0.996). Detection of DNA at a concentration as low as the equivalent of a single cell showed that the assay was sensitive enough to detect a single cell of P. olseni. The estimated number of P. olseni cells was similar to the original cell concentrations, indicating the reliability of P. olseni quantification by real-time PCR. Accordingly, the designed primers and probe may be used for the rapid detection and quantification of P. olseni from clam tissue, environmental water, and sediment samples.

Effect of pH on the ATP-sensitive $K^+$ Channel in Aortic Smooth Muscle Cells from Rats

  • Kim, Se-Hoon;Kim, Il-Su;Kim, Hoe-Suk;Jeon, Byeong-Hwa;Chang, Seok-Jong
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.5
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    • pp.555-563
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    • 1997
  • The effects of pH on $K^+$ currents were investigated in single smooth muscle cells isolated from the thoracic aorta of Wistar-Kyoto rats. Whole-cell $K^+$ currents were recorded in the conventional configuration of the voltage-clamp technique. Pinacidil (10uM) activated the whole-cell current and the pinacidil-activated current was completely inhibited by glibenclamide (10uM) , an inhibitor of ATP-sensitive $K^+$ channel ($K_{ATP}$ channel). Pinacidil-activated current was reversed at near the $K^+$ equilibrium potential. This current was time- and voltage-independent and reduced by elevating intracellular ATP. Pinacidil-activated current was reduced by lowering the external pH. However, alteration of internal pH has controversial effects on pinacidil-activated current. When the single cell was dialyzed with 0.1 mM ATP, alteration of internal pH had no effect on pinacidil-activated $K^+$ current. In the contrast, when the single cell was dialyzed with 3 mM ATP, pinacidil-activated current was increased by lowering internal pH. Our results suggest that $K^+$ channel activated by pinacidil may be $K_{ATP}$ channel and internal $H^+$ may reduce the inhibitory effect of ATP on $K_{ATP}$ channel.

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Use of the In Vivo Single-cell Gel Electrophoresis Assay for Evaluating Genotoxicity in Clam (Single-cell Gel Electrophoresis Assay에 의한 대합에서의 In Vivo 유전독성 평가)

  • Kim Il-Yang;Hyun Chang-Kee
    • Toxicological Research
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    • v.20 no.3
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    • pp.225-232
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    • 2004
  • The suitability of the single cell gel electrophoresis (SCGE) assay as a test for the monitoring of genotoxicity of aquatic environment was evaluated. The SCGE assay was employed to detect DNA damage induced in clam (Spisula sachalinensis) exposed to a direct mutagen, N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) or an indirect mutagen, benzo[a]pyrene (B[a]P). The cells of gill and digestive glands were isolated from clam by homogenization, which was the optimized cell dissociation method, and the level of DNA damage was assessed and expressed as mean tail length. In the gill cells, significant dose- and time-dependent increase was observed in the mean tail length at the concentration from 0.01 to 0.5 ppm MNNG for 96 h. The linear correlation between relative dam-age index (RDI) values was suggested to provide criteria of genotoxicity monitoring for direct acting mutagen. The dose- and time-dependent responses of the digestive glands cells were less sensitive than those of the gill cells. In contrast, the genotoxic response resulting from the exposure of 0.01~1.0 ppm B[a]P to clam revealed a higher sensitivity in the digestive glands cells than the gill cells. The comparison between the time profiles of genotoxic responses in clam and carp, the latter had been obtained in our previous study, indicated that the metabolism of genotoxic compounds in the two aquatic organisms were quite different each other. We conclude that the SCGE assay has the potential as a screening test for routine genotoxicity monitoring of aquatic organisms because of its higher sensitivity and simplicity.