• BMB Reports
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• 제37권2호
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• pp.239-245
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• 2004

We have previously demonstrated that the redox reactant pyruvate prevents apoptosis in the oxidant model of bovine pulmonary artery endothelial cells (BPAEC), and that the anti-apoptotic mechanism of pyruvate is mediated in part via the mitochondrial matrix compartment. However, cytosolic mechanisms for the cytoprotective feature of pyruvate remain to be elucidated. This study investigated the pyruvate protection against endothelial cytotoxicity when the glycolysis inhibitor 2-deoxy-D-glucose (2DG) was applied to BPAEC. Millimolar 2DG blocked the cellular glucose uptake in a concentration- and time-dependent manner with >85% inhibition at $\geq$5 mM within 24 h. The addition of 2DG evoked BPAEC cytotoxicity with a substantial increase in lipid peroxidation and a marked decrease in intracellular total glutathione. Exogenous pyruvate partially prevented the 2DG-induced cell damage with increasing viability of BPAEC by 25-30%, and the total glutathione was also modestly increased. In contrast, 10 mM L-lactate, as a cytosolic reductant, had no effect on the cytotoxicity and lipid peroxidation that are evoked by 2DG. These results suggest that 2DG toxicity may be a consequence of the diminished potential of glutathione antioxidant, which was partially restored by exogenous pyruvate but not L-lactate. Therefore, pyruvate qualifies as a cytoprotective agent for strategies that attenuate the metabolic dysfunction of the endothelium, and cellular glucose oxidation is required for the functioning of the cytosolic glutathione/NADPH redox system.

• 한국가축번식학회지
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• 제19권3호
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• pp.161-169
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• 1995

To investigate the metabolism of various substrates in preimplantation bovine embryos, uptake of glucose and pyruvate, and lactate production were measured in single IVF-derived bovine embryos by a non-invasive method. When the embryos were incubated for 5 h in culture medium supplemented with 1 mM glucose and 0.4mM pyruvate as substrates at each developmental stage, glucose uptake was increased with more advanced developmental stages while pyruvate uptake was decreased. Total lactate producton of 2-cell embryos was significantly higher than that of blastocysts (p<0.05). Both of glucose uptake and lactate production in normal morulae produced in vitro was significantly high compared to the degenerated embryos(p<0.05). The results obtained in the study suggest that pyruvate as an exogenous substrate may be support in bovine embryos until 8-cell stage, whereas glucose may be effective as an energy source after morula stage. In addition, it was proven thatlactate was not effective as an energy source in preimplantation development of IVF-derived bovine embryos.

• 원예과학기술지
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• 제33권5호
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• pp.763-771
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• 2015

작물의 구성성분이나 생리활성은 재배지역의 자연환경에 영향을 많이 받는다. 제주 동부와 서부 마늘 재배지역의 토양분석 및 생산된 마늘의 성분분석을 실시하였다. 토양 및 마늘의 대량원소는 동부지역에서 높게 나타났으며 미량원소는 서부지역에서 높게 나타났다. 또한 알리인과 총 페놀 함량은 서부지역에서 높았으며 알리신, 총 피루베이트산, 총 플라보노이드 함량 및 ${\alpha}$-glucosidase 저해활성은 동부지역에서 높게 나타나 지역간 차이가 나타남을 알 수 있었다. 본 연구는 제주에서 재배지역에 따른 마늘 성분의 차이를 이해하는데 도움을 줄 것으로 사료된다.

• BMB Reports
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• 제31권3호
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• pp.258-263
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• 1998

Acetolactate synthase (ALS) is the common enzyme in the biosynthetic pathways leading to leucine, valine, and isoleucine. Tobacco ALS was expressed in E. coli and purified to homogeneity. The recombinant tobacco ALS was inactivated by thiol-specific reagents, N-ethylmaleimide (NEM) and 5,5'-dithio-bis-(2-nitrobenzoic acid) (DTNB). Inactivation of the ALS by NEM followed pseudo-first order kinetics and was first order with respect to the modifier. The substrate pyruvate protected the enzyme against the inactivation by NEM and DTNB. Extrapolation to complete inactivation of the enzyme by DTNB showed modification of approximately 2 out of 4 total cysteinyl residues (or 2 cysteinyl and 1 cysteinyl residues), with approximately 1 residue protected by pyruvate. The tobacco ALS was also inactivated by the tryptophanspecific reagent, N-bromosuccinimide (NBS), and was similarly protected by pyruvate. The kinetics of the inactivation was first-order with respect to NBS. The present data suggest that cysteinyl and tryptophanyl residues play a key role in the catalytic function of the enzyme.

• 대한의생명과학회지
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• 제21권4호
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• pp.241-247
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• 2015

Lactate and ketone bodies are considered biological markers for ketosis and several inherited metabolic disorders. In the current study, the specific ratios of lactate and ketone bodies as analytical tools for differential diagnosis of various lactic acidosis were devised. The study included a protein precipitation step following tert-butyldimethylsilyl derivatisation. Total run time was approximately 30 min including sample preparation and GS/MS analysis. The limits of detection were below 0.1 pg/mL over the targeted 4 analytes. The calibration curve was linear over the concentration range of $0.001{\sim}250{\mu}g/mL$ for pyruvate, beta-hydroxybutyrate, and acetoacetate ($R^2$ > 0.99). Inter-day accuracy and precision were 87.7~94.8% with RSD of 2.5~5.7% at 2 levels. Absolute recoveries (%) of target analytes were 87.0~98.4%. The method was validated for the quantification of lactate and ketone bodies for differentiation of lactic acidosis.

• Investigative Magnetic Resonance Imaging
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• 제26권2호
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• pp.125-134
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• 2022

Purpose: The aim of this study was to investigate the change in signal sensitivity over different acquisition start times and optimize the scanning window to provide the maximal signal sensitivity of [1-¹³C]pyruvate and its metabolic products, lactate and alanine, using spatially localized hyperpolarized 3D ¹³C magnetic resonance spectroscopic imaging (MRSI). Materials and Methods: We acquired 3D ¹³C MRSI data from the brain (n = 3), kidney (n = 3), and liver (n = 3) of rats using a 3T clinical scanner and a custom RF coil after the injection of hyperpolarized [1-¹³C]pyruvate. For each organ, we obtained three consecutive 3D ¹³C MRSI datasets with different acquisition start times per animal from a total of three animals. The mean signal-to-noise ratios (SNRs) of pyruvate, lactate, and alanine were calculated and compared between different acquisition start times. Based on the SNRs of lactate and alanine, we identified the optimal acquisition start timing for each organ. Results: For the brain, the acquisition start time of 18 s provided the highest mean SNR of lactate. At 18 s, however, the lactate signal predominantly originated from not the brain, but the blood vessels; therefore, the acquisition start time of 22 s was recommended for 3D ¹³C MRSI of the rat brain. For the kidney, all three metabolites demonstrated the highest mean SNR at the acquisition start time of 32 s. Similarly, the acquisition start time of 22 s provided the highest SNRs for all three metabolites in the liver. Conclusion: In this study, the acquisition start timing was optimized in an attempt to maximize metabolic signals in hyperpolarized 3D ¹³C MRSI examination with [1-¹³C] pyruvate as a substrate. We investigated the changes in metabolic signal sensitivity in the brain, kidney, and liver of rats to establish the optimal acquisition start time for each organ. We expect the results from this study to be of help in future studies.

• 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.

• The Korean Journal of Physiology
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• 제1권1호
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• pp.33-41
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• 1967

The metabolic patterns of C-1 and C-6-carbon atoms of glucose were observed in the tissue homogenates of the Ehrlich ascites tumor tissue which was incubated for 3 hours in the Dubnuff metabolic shaking incubator. $C^{14}-1-and\;C^{14}-6-glucose$ were used as tracers. The glucose media in which tissue homogenate was incubated was kept at a concentration of 200mg% glucose of carrier and appropriate amount of $C^{14}-1-or\;C^{14}-6-tracer$. At the end of 3 hour incubation, respiratory $CO_2$ samples trapped by alkaline which is placed in the tenter well of incubation flask were analyzed for the total $CO_2$ production rates and their radioactivities. The tissue homogenate samples after incubation were analyzed for their concentrations of glucose, lactate, pyruvate and glycogen and calculations were made on the glucose consumption rate, pyruvate and lactate accumulation rates. The following results were obtained. Data obtained in each group are as follows: 1. In the tissue homogenate, which was incubated with $C^{14}-1-glucose as a substrate, total $CO_2$ production rate averaged $19.0{\pm}5.0{\mu}M/hr/gm$ and the mean specific activity of respiratory $CO_2$ was $840{\pm}296\;cpm/mgC.$ Relative specific activity (RSA) which means the fraction of $CO_2$ derived from medium $C^{14}-1-glucose$ to total $CO_2$ production rate was calculated by ratio of SA of respiratory $CO_2$ and medium $C^{14}-1-glucose.$ RSA was $14.3{\pm}5.0%,$ Accordingly actual $CO_2$ production rate from medium $C^{14}-1-glucose$ showed a mean value of $2.79{\pm}1.35\;{\mu}m$ of which amount was equivalent to the mean value of total glucose consumption rate $(RGDco_2)$, namely, $5.1{\pm}1.3%.$ Lactate and pyruvate appearance rates averaged $7.13{\pm}1.26\;and\;0.21{\pm}0.02{\mu}M/hr/gm,$ respectively. Assuming that these 3 carbon compounds appeared in the medium were derived from glucose, calculations were made that relative glucose disappearance rate into lactate $(RGD_L)$ was $38.0{\pm}5.4%\;and\;RGD_P$ was $1.23{\pm}0.03%.$ Therefore, about 43.3% of the total glucose consumed were accounted for by conversion into the respiratory $CO_2$, lactate and pyruvate. 2. In the second group, which was incubated with $C^{14}-1-glucose$ as a substrate, glucose consumption rate, lactate and pyruvate appearance rates showed almost the same order as the values of the $C^{14}-1-glucose$ substrate group. However, RSA was remarkably decreased showing a mean value of $1.02{\pm}0.13%.$ This fact means that the C-6 carbon of glucose take the minor part in the oxidative metabolism of glucose. The glycogen level in both substrate tissue homogenate showed less than 0.3% of tissue weight. These low value suggested that there was an inhibition of carbohydrate synthesis in the Ehrlich ascites tumor tissue. 3. The catabolic pathway of glucose in the tumor tissue were analyzed on the basis of Bloom's principle from the values of RSA. It was found that in the tumor tissue more than 90% of $CO_2$ derived from glucose were oxidized via the alternate pathway other than principal EMP-TCA cycle such as hexose monophosphate pathway (HMP). From the data described above, it was assumed that in the Ehrlich tumor tissue anaerobic glycolysis proceeds normally although, the oxidation of products of anaerobic glycolysis via the TCA cycle is inhibited resulting in the accumulation of lactate and almost all of oxidative energy from glucose is released by oxidative pathway such as HMP.

• The Korean Journal of Physiology
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• 제3권1호
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• pp.11-18
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• 1969

Tissue homogenates of 10 kinds of human cancer tissues were incubated in medium containing either one of $C^{14}-1,\; C^{14}-2,\;or\; C^{14}-3-lactate $ as a substrate in order to observe the oxidative pathway of lactate in cancer tissues. Lactate concentration in incubation medium was maintained at 50 mg%. At the end of incubation period, gas samples and incubation media were analyzed for total $CO_2$ production rates, radioactivities of respiratory $CO_2$, lactate uptake rates and pyruvate appearance rates. The following results were obtained. 1. Lactate uptake rates in all of cancer tissues examined were less than $2.5\;{\mu}M/hr/gm$ and much lower than those in normal tissues. 2. In the 10 kind of human cancer tissues, total $CO_2$ production rates were less than $10\;{\mu}M/hr/gm$, in all cases. These lower values impressed that oxidative metabolism in tumor tissues generally inhibited as compared with that in normal tissue. On the other hand, fractions of $CO_2$ derived from lactate to total $CO_2$ production rates were less than 15% except one case These facts showed that oxidation of lactate into $CO_2$ was greatly inhibited in tumor tissues. 3. Respiratory $CO_2$ yields from C-1 carbon of lactate in various cancer tissues were mean of 77.7% of total $CO_2$ yield from lactate and $CO_2$ yields from C-2 and C-3 carbon of lactate were mean of 9.1% and 12.6% respectively. These facts showed that carboxyl carbon of lactate oxidized more easily than ${\alpha}\;and\;{\beta}$ carbon of lactate. 4. In 10 kinds of cancer tissues, fractions of disappeared lacteate from media into $CO_2$ and pyruvate, which expressed as RLD $co_2$ and RLDpy respectively, were about 5% in except 3 cases and less than 3% except one case. These fact showed that almost of disappeared lactate from media were degraded into compounds other than $CO_2$ and pyruvate. From the above date, it was suggested that in the oxidative pathway of lactate in cancer tissues $CO_2$ was easily Produced from carboxyl carbon of lactate by oxidative decarboxylation as in the normal tissue, and further oxidation of 2 carbon unit via TCA cycle was inhibited.

• 농업생명과학연구
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• 제46권5호
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• pp.91-100
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• 2012

국내 마늘의 품종과 관련한 연구는 산지에 따른 비교로 한정되어 있어 마늘의 성분 및 특성에 대한 기초자료를 확보하고자 품종별 마늘 시료 12종을 대상으로 이화학적 특성과 항균활성을 분석하였다. 품종별 마늘의 구폭은 캘리포니아 만생종(60.44mm, 53.73g)이 가장 크고 무거웠으며, 창녕종(44.04mm, 25.15g)이 가장 작고 가벼웠다. 품종별 마늘의 표면색 중 L값은 84.13~90.56, a값은 -1.10~0.77, b값은 18.24~26.61의 범위였다. 전단가는 캘리포니아 조생종이 가장 낮았으며, 94-12-2종은 $4211.35cm/kg^2$ 으로 타 시료에 비하여 전단가가 가장 높았다. 가용성 고형분 함량은 6.40~11.33 %brix의 범위였는데, 창녕종에서 유의적으로 높았다. 품종별 마늘의 pH는 5.57~6.53의 범위였다. 총 thiosulfinate 함량은 이태리종이 93.23 mM/g로 가장 낮았고, 캘리포니아 만생종이 146.05 mM/g로 가장 높았다. 총 pyruvate 함량은 유고종($162.50{\mu}M/g$)에서 가장 높았고, 캘리포니아 조생종($147.41{\mu}M/g$)에서 가장 낮았다. 품종 별 마늘의 추출물로 항균활성을 측정한 결과, Streptococcus mutans균에서 활성이 가장 낮았으며, Escherichia coli균에 대한 항균 활성이 높았다.