• Journal of Microbiology and Biotechnology
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• 제30권5호
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• pp.753-761
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• 2020

To determine the role of pyruvate dehydrogenase complex (PDHC) in Klebsiella pneumoniae, the growth and metabolism of PDHC-deficient mutant in glycerol-based medium were analyzed and compared with those of other strains. Under aerobic conditions, the PDHC activity was fourfold higher than that of pyruvate formate lyase (PFL), and blocking of PDHC caused severe growth defect and pyruvate accumulation, indicating that the carbon flux through pyruvate to acetyl coenzyme A mainly depended on PDHC. Under anaerobic conditions, although the PDHC activity was only 50% of that of PFL, blocking of PDHC resulted in more growth defect than blocking of PFL. Subsequently, combined with the requirement of CO₂ and intracellular redox status, it was presumed that the critical role of PDHC was to provide NADH for the anaerobic growth of K. pneumoniae. This presumption was confirmed in the PDHC-deficient mutant by further blocking one of the formate dehydrogenases, FdnGHI. Besides, based on our data, it can also be suggested that an improvement in the carbon flux in the PFL-deficient mutant could be an effective strategy to construct high-yielding 1,3-propanediol-producing K. pneumoniae strain.

• 생약학회지
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• 제20권1호
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• pp.32-36
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• 1989

Effects of Lycium chinensis, Epimedium koreanum and tuguaconitine which is isolated from Aconitum sibiricum on primary culture chicken embryonic brain cells were studied by microscopic observation and determined of the activity of pyruvate dehydrogenase complex(PDHC). Brain cells were prepared from the brain of 10-day-old chicken embryo and cultured with a medicine consisted of 90% Dulbecco's Modified Eagle Medium(DMEM) and 10% horse serum. It was observed that all substances studied seemed to show the tendency to stimulate the neurite outgrowth of brain cells which were cultured with a deficient medium under microscopic observation. The activity of PDHC in brain cells cultured with a deficient medium was increased by Lysium chinensis and Epimedium koreanum. However, tuguaconitine had not influence on the activity of PDHC.

• 약학회지
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• 제33권1호
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• pp.39-45
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• 1989

Effects of dammarane glycosides of Panax ginseng on primary cultured chicken embryonic brain cells were studied by microscopic observation and determination of the activity of pyruvate dehydrogenase complex (PDHC). Brain cells were prepared from the brain of 10-day-old chicken embryo and cultured with either a standard medium consisted of 85% Dulbecco's Modified Eagle Medium (DMEM), 10% horse serum and 5% chicken embryonic extracts or a deficient medium consisted of 90% DMEM and 10% horse serum. It was observed that dammarane glycosides of Panax ginseng seemed to show the tendency to stimulate the neurite outgrowth of brain cells which were cultured with a deficient medium under microscopic observation. The activity of PDHC in brain cells cultured with a deficient medium was increased by dammarane glycosides of Panax ginseng.

• Archives of Pharmacal Research
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• 제14권4호
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• pp.325-329
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• 1991

Effects of Lycii Fructus on primary cultured chicken embryonic brain cells were studied by microscopic observation, determination of the activity of pyruvate dehydrogenase complex (PDHC), and syntheses of protein, RNA and DNA. The brain cells were prepared from the brains or 10-day-old chicken embryos and cultured with a deficient medium. The activity of PDHC in the brain cells cultured with a deficient medium was increased to 1.8 times by the addition of $30\;{\mu}g/ml$ of the total methanol extract of Lycii Fructus. To seek the active fraction, total methanol extract was further fractionated by the polarity. The survival rate of neuronal cells was significantly increased by the addition of $100\;{\mu}g/ml$ of the buthanol or aqueous fraction. At this concentration, the significant increase of the syntheses of protein and RNA, but not of DNA, indicates that the fractions may act on the neuronal cells which are known to be non-dividing cells.

• 약학회지
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• 제34권5호
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• pp.365-373
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• 1990

The effects of the protein fraction of Panax ginseng on primary cultured chicken embryonic brain cells and DRG cultured with a deficient medium were studied. The protein fraction was further fractionated into four groups according to the molecular weight; larger than 10,000 dalton(fraction A), between 5,000 and 10,000 daltons(fraction B), between 1,000 and 5,000 daltons(fraction C), between 500 and 1,000 daltons(fraction D). All four protein fractions at the concentration of $100\;{\mu}g/ml$ significantly increased the number of the brain cells which promoted the neurite outgrowth. The activity of PDHC in the brain cells was elevated significantly by the protein fraction B at the concentration of $100\;{\mu}g/ml$. It was noted that $100\;{\mu}g/ml$ protein fraction C and D significantly enhanced the synthesis of protein in the brain cells. At the concentration of $100\;{\mu}g/ml$, the protein fraction B enhanced RNA synthesis and the protein fraction A significantly enhanced DNA synthesis in the brain cells. The protein fractions B, C, and D significantly promoted the neurite outgrowth of DRG at the concentration of $100\;{\mu}g/ml$.

• 생약학회지
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• 제21권1호
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• pp.92-99
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• 1990

Effects of the methanol extract of Bupleuri Radix on primary cultured chicken embryonic brain cells, dorsal root ganglia (DRG) and rat hepatocytes were studied. The methanol extract of Bupleuri Radix at the concentration ranging from $10{\;}{\mu}g/ml\;to\;100{\;}{\mu}g/ml$ significantly recovered the cytotoxicity of rat hepatocytes induced by the treatment of galactosamine; at the concentration of $100\;{\mu}g/ml$, values of GOT and GPT in the culture medium were reduced by the 60% and 75%, respectively of those in the absence of the methanol extract of Bupleuri Radix. The addition of the methanol extract of Bupleuri Radix. into chicken embryonic brain cells which were cultured with a deficient medium significantly increased the number of cells promoting the neurite outgrowth. However, the methanol extract of Bupleuri Radix showed no effect on the activities of PDHC and acetylcholinesterase in primary cultured brain cells.

• 대한유전성대사질환학회지
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• 제11권1호
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• pp.52-73
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• 2011

Since we have started organic acid analysis on Jul. 1997, we have been collecting data about organic acidemias in Korea. The data presented here is our 3 years experience in organic acid analysis. We have collected 712 samples from major university hospitals all over the Korea, large enough for relatively accurate incidence of organic acid disorders. We are using solvent extraction method with ethylacetate, MSTFA for derivatization and quantitation of 83 organic acids simultaneously. Out of 712 patients sample, 498 patients sample (70%) showed no evidence of organic acid abnormalities. Out of 214 remaining samples we have found very diverse disorders such as methylmalonic aciduria(6), propionic aciduria (10), biotinidase deficiency (6), maple syrup urine disease (3), isovaleric aciduria (4), tyrosinemia type II (4), tyrosinemia type IV (1), glutaric aciduria type I (1), glutaric aciduria type II (22), 3-methylglutaconic aciduria type I (3), 3-methylglutaconic aciduria type III (7), HMG-CoA lyase deficiency (1), hyperglyceroluria (2), cytosolic 3-ketothiolase deficiency (55), mitochondrial 3-ketothiolase deficiency (3), 3-hydroxyisobutyric aciduria (2), L-2-hydroxyglutaric aciduria (2), fumaric aciduria (2), lactic aciduria with combined elevation of pyruvate (most likely PDHC deficiency) (28), lactic aciduria without combined elevation of pyruvate (most likely mitochondrial respiratory chain disorders) (35), SCAD deficiency (3), MCAD deficiency (1), 3-methylcrotonylglycineuria (1), orotic aciduria (most likely urea cycle disorders) (7) and 2-methylbranched chain acyl-CoA dehydrogenase deficiency (1). In conclusion, though the incidence of indivisual organic acidemia is low, the incidence of overall organic acidemia is relatively high in Korea. Most of the patients showed some signs of neurological dysfunction. In other words, organic acid analysis should be included in the diagnostic work up of all neurological dysfunctions.

• Clinical and Experimental Pediatrics
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• 제49권3호
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• pp.258-267
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• 2006

목 적 : 국내의 유기산 대사 이상 질환들의 상대적 발병 빈도와 임상양상을 알아보기 위하여 유기산 정량 분석이 가능하여진 1997년 7월부터 2005년 5월까지 8년간 전국에 있는 대학병원에서 의뢰된 1,787명의 환자의 검체를 분석한 결과를 3년 전 발표한 연구결과와 비교하여 정리하였다. 방 법 : 1997년 7월 1일부터 2005년 5월 31일까지 약 8년 동안 본원 임상유전학연구실로 분석이 의뢰되었던 1,787명 환자의 검체들의 분석결과를 신생아기(생후 2개월까지), 영아기(2개월 이후 2세까지), 유소아기(2세 이후 12 세까지) 및 청소년기 이후(12세 이후)로 분류하였고, 환자들의 주요증상 및 소견을 분류하였다. 유기산 분석은 83종의 유기산을 gas chromatography와 mass spectrometry를 이용하여 정량 분석하였으며, 특징적인 양상으로 진단하였다. 결 과 : 총 28가지 질환 470명의 환자를 이 기간 동안에 진단할 수 있었으며, 가장 빈도가 높았던 질환들은 cytosolic 3-ketothiolase 결핍증, 사립체의 호흡 연쇄 효소의 이상 질환군(mitochondrial respiratory chain disorders), PDHC 결핍증, mitochondrial 3-ketothiolase 결핍증, glutaric aciduria type II biotinidase 결핍증, 메틸말론산뇨증(methylmalonic aciduria), 프로피온산뇨증(propionic aciduria) 등이었다. 결 론 : 유기산 대사 장애 질환들은 각각의 발병 빈도는 낮으나 국내에서도 다양한 질환들이 발병되고 있으며, 이들 질환들은 대부분의 경우 신경학적 이상 소견을 보이고 있으므로 신경학적 이상 소견을 보이는 경우는 감별진단에 포함시켜야 된다. 3년전 Choi 등이 발표한 연구결과와 비교하여 새로운 질환의 출현은 없었고 빈도가 높은 질환의 발병빈도의 차이도 보이지 않았다. 3년 전의 연구당시 짧은 연구기간으로 인한 오차의 가능성을 염려하였으나 8년 동안의 긴 연구기간 동안 비슷한 상대적 발병빈도를 보여 이러한 질환들이 우리나라에서 다수 발병되는 것으로 판단되며 주의가 필요하다고 판단된다.