• Title/Summary/Keyword: enzymatic analysis

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Three Cases of Mitochondrial Disorders in the Neonatal Period (신생아기에 진단된 미토콘드리아 질환 3례)

  • Kim, Yoon-Hee;Lee, Young-Mock;Namgung, Ran;Kim, Jeong-Eun;Lee, Soon-Min;Park, Kook-In;Kim, Se-Hoon;Lee, Jin-Sung
    • Neonatal Medicine
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    • v.17 no.2
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    • pp.254-261
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    • 2010
  • Little is known about neonatal mitochondrial disease, though mitochondrial metabolic disorders may often present in the neonatal period because of the high energy requirement of neonate. In newborn period, common presentations are not specific and the disease course may be rapid and fatal. In this study, we report three cases of neonatal mitochondrial disease. The first case was strongly suspected because of sudden seizure and mental change with severe lactic acidosis, and multiorgan failure. Plasma lactate/pyruvate (L/P) ratio was increased to 55.6 with marked lactic aciduria and increased plasma alanin up to 2,237 nmol/mL. In the second patient, a peritoneal dialysis was performed for acute adrenal and renal failure, but metabolic acidosis persisted. Plasma L/P ratio was increased to 23.9, and MRC I (mitochondrial respiratory chain defect) was diagnosed through the enzymatic analysis of the muscles. The third case showed repetitive episode of lactic acidosis during the first two months of life, hypotonia, failure to thrive and feeding difficulties. We found markedly increased cerebrospinal fluid L/P ratio up to 57 though plasma L/P ratio(19.4) was borderline with increased plasma lactate. The lactate peak was prominent in brain magnetic resonance spectroscopy (MRS). MRC II was confirmed through muscle biopsy. Plasma lactate level and lactate peak of brain MRS were normalized after conservative treatment.

Screening of Biodegradable Function of Indigenous Ligno-degrading Mushroom Using Dyes

  • Jang, Kab-Yeul;Cho, Soo-Muk;Seok, Soon-Ja;Kong, Won-Sik;Kim, Gyu-Hyun;Sung, Jae-Mo
    • Mycobiology
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    • v.37 no.1
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    • pp.53-61
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    • 2009
  • The process of biodegradation in lingo-cellulosic materials is critically relevant to biospheric carbon. The study of this natural process has largely involved laboratory investigations, focused primarily on the biodegradation and recycling of agricultural by-products, generally using basidiomycetes species. In order to collect super white rot fungi and evaluate its ability to degrade lingo-cellulosic material, 35 fungal strains, collected from forests, humus soil, livestock manure, and dead trees, were screened for enzyme activities and their potential to decolorize the commercially used Poly-R 478 dye. In the laccase enzymatic analysis chemical test, 33 white rot fungi and 2 brown rot fungi were identified. The degradation ability of polycyclic aromatic hydrocarbons (PAHs) according to the utilized environmental conditions was higher in the mushrooms grown in dead trees and fallen leaves than in the mushrooms grown in humus soil and livestock manure. Using Poly-R 478 dye to assess the PAH-degradation activity of the identified strains, four strains, including Agrocybe pediades, were selected. The activities of laccase, MnP, and Lip of the four strains with PAH-degrading ability were highest in Pleurotus incarnates. 87 fungal strains, collected from forests, humus soil, livestock manure, and dead trees, were screened for enzyme activities and their potential to decolorize the commercially used Poly-R 478 dye on solid media. Using Poly-R 478 dye to assess the PAHdegrading activity of the identified strains, it was determined that MKACC 51632 and 52492 strains evidenced superior activity in static and shaken liquid cultures. Subsequent screening on plates containing the polymeric dye poly R-478, the decolorization of which is correlated with lignin degradation, resulted in the selection of a strain of Coriolus versicolor, MKACC52492, for further study, primarily due to its rapid growth rate and profound ability to decolorize poly R-478 on solid media. Considering our findings using Poly-R 478 dye to evaluate the PAH-degrading activity of the identified strains, Coriolus versicolor, MKACC 52492 was selected as a favorable strain. Coriolus versicolor, which was collected from Mt. Yeogi in Suwon, was studied for the production of the lignin-modifying enzymes laccase, manganese-dependent peroxidase (MnP), and lignin peroxidase (LiP).

Comparison between Single and Co-culture of Adipocyte and Muscle Cell Lines in Cell Morphology and Cytosolic Substances (지방과 근육 세포주의 단독 및 공동배양을 통한 세포형태학 및 세포물질 비교 연구)

  • Choi, Chang-Weon;Cho, Won-Mo;Yeon, Seong-Heum;HwangBo, Soon;Song, Man-Kang;Park, Sung-Kwon;Baek, Kyung-Hoon
    • Journal of Animal Science and Technology
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    • v.54 no.2
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    • pp.103-109
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    • 2012
  • Present study was performed to investigate the effect of single and co-culture of adipocyte and muscle cell lines on cell differentiation. 3T3-L1 (adipocyte) and L6 (muscle) cell lines were single-cultured on the condition of 10% fetal bovine serum (FBS)/Dulbeco's modified eagle's medium (DMEM) for 48 h followed by culture within 5% FBS/DMEM as a growth media. Then, the growth media was replaced by differentiation media composed of 2% FBS/DMEM without additives in single- or co-culture of the 3T3-L1 and the L6 cells to induce differentiation of both cell types. In co-culture system, the 3T3-L1 and the L6 cells were grown in separated places by being seeded on a $0.4{\mu}m$ insert membrane and on the bottom of 6 well plate, respectively. Cell differentiation was measured using morphological investigation and cytosolic analysis of glycerol-3-phosphate dehydrogenase (GPDH; for 3T3-L1) and creatine kinase (CK; for L6). Based on the GPDH results, the presence of L6 cells did not stimulate 3T3-L1 differentiation showing more differentiation of 3T3-L1 cells in the single-culture compared to the co-culture condition. In contrast, 3T3-L1 cells in the co-culture promoted differentiation of L6 cells. Enzymatic analysis supported this result showing that 3T3-L1 cells showed statistically (P<0.05) higher GPDH activity in the single-culture than the co-culture, whereas CK results of L6 cells were vice versa (P<0.05). Overall, present results may indicate that co-culture system is more reliable and precise technique compared to single-culture. Further studies on several co-culture trials including different media conditions, supplementation of differentiating substances, molecular biological analysis, etc. should be required to obtain practical and fundamental mass data.

Gelatinization Properties of Starch Dough with Moisture Content, Heating Temperature and Heating Time (수분함량, 가열온도 및 가열시간에 따른 전분 반죽의 호화특성)

  • Lee, Boo-Yong;Lee, Chang-Ho;Lee, Cherl-Ho
    • Korean Journal of Food Science and Technology
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    • v.27 no.3
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    • pp.428-438
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    • 1995
  • The gelatinization properties of corn and waxy corn starch doughs were examined at various moisture contents, heating temperatures and heating times. The onset temperatures of gelatinization with 1% CMC using Brabender Amylograph were $64^{\circ}C$ for both corn and waxy corn starch. In the gelatinization properties using DSC, onset temperature$(T_o)$, maximum peak temperature$(T_p)$, completion temperature$(T_c)$ and enthalpy of the corn starch were $68.15^{\circ}C,\;74.01^{\circ}C,\;85.65^{\circ}C$ and $3.2\;cal/gram$ respectively. While those of the waxy corn starch were $68.24^{\circ}C,\;75.43^{\circ}C,\;93^{\circ}C$ and $4.2\;cal/gram$ respectively. In enzymatic analysis, when the moisture content increased from 36% to 52% and heating temperature from $60^{\circ}C$ to $100^{\circ}C$, the gelatinization degree of starch dough increased from about 10% to about 62%. The gelatinization degree of waxy corn starch dough was $15{\sim}20%$ higher than that of corn starch dough under the same gelatinization conditions. The regression equations of gelatinization degree (Y) of starch dough in the range of $36{\sim}52%$ moisture content $(X_1)\;60{\sim}100^{\circ}C$ heating temperature $(X_2)\;and\;0{\sim}2.0$ min heating time $(X_3)$ were examined using response surface analysis. The regression equation of corn starch dough was: $Y=28.659+8.638\;X_}+15.675\;X_2+7.770\;X_3-1.620\;{X_1}^2+10.790\;X_1X_2-4.220\;{X_2}^2+0.510\;X_1X_3+1.980\;X_2X_3-6.850\;{X_3}^2\;(R^2=0.9714)$ and that of waxy corn starch dough was: $Y=32.617+12.535\;X_1+20.470\;X_2+8.608\;X_3+4.093\;{X_1}^2+13.550\;X_1X_2-4.467\;{X_2}^2+1.560\;X_1X_3+2.160\;X_2X_3-9.527\;{X_3}^2$\;(R^2=0.9621)$. As the moisture content, heating temperature and heating time increased, the reaction rate constant(k) of gelatinization increased. The greatest reaction rate constant was observed at initial 0.5 min heating time of 1st gelatinization stage. At the heating temperature of $90^{\circ}C$, gelatinization of starch dough was completed almost in the initial 0.5 min heating time. The reaction rate constant of waxy corn starch dough was higher than that of corn starch dough under the same gelatinization conditions. At the 52% moisture content, the regression equation between reaction rate constant(k) and heating temperature(T) for corn starch dough was $log\;k=11.1140-4.1226{\times}10^3(1/T)$ (r=-0.9520) and that of waxy corn starch dough was $log\;k=10.1195-3.7090{\times}10^3(1/T)$ (r=-0.9064).

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