• Investigative Magnetic Resonance Imaging
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• 제21권1호
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• pp.1-8
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• 2017

Purpose: To investigate the exchange and redistribution of hyperpolarized $^{13}C$ metabolites between different pools by temporally analyzing the relative fraction of dual $T_2{^*}$ components of hyperpolarized $^{13}C$ metabolites. Materials and Methods: A dual exponential decay analysis of $T_2{^*}$ is performed for [1-$^{13}C$] pyruvate and [1-$^{13}C$] lactate using nonspatially resolved dynamic $^{13}C$ MR spectroscopy from mice brains with tumors (n = 3) and without (n = 4) tumors. The values of shorter and longer $T_2{^*}$ components are explored when fitted from averaged spectrum and temporal variations of their fractions. Results: The $T_2{^*}$ values were not significantly different between the tumor and control groups, but the fraction of longer $T_2{^*}$ [1-$^{13}C$] lactate components was more than 10% in the tumor group over that of the controls (P < 0.1). The fraction of shorter $T_2{^*}$ components of [1-$^{13}C$] pyruvate showed an increasing tendency while that of the [1-$^{13}C$] lactate was decreasing over time. The slopes of the changing fraction were steeper for the tumor group than the controls, especially for lactate (P < 0.01). In both pyruvate and lactate, the fraction of the shorter $T_2{^*}$ component was always greater than the longer $T_2{^*}$ component over time. Conclusion: The exchange and redistribution of pyruvate and lactate between different pools was investigated by dual component analysis of the free induction decay signal from hyperpolarized $^{13}C$ experiments. Tumor and control groups showed differences in their fractions rather than the values of longer and shorter $T_2{^*}$ components. Fraction changing dynamics may provide an aspect for extravasation and membrane transport of pyruvate and lactate, and will be useful to determine the appropriate time window for acquisition of hyperpolarized $^{13}C$ images.

• 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.

• Korean Journal of Radiology
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• 제25권5호
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• pp.459-472
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• 2024

Hyperpolarized (HP) carbon-13 (¹³C) MRI represents an innovative approach for noninvasive, real-time assessment of dynamic metabolic flux, with potential integration into routine clinical MRI. The use of [1-¹³C]pyruvate as a probe and its conversion to [1-¹³C]lactate constitute an extensively explored metabolic pathway. This review comprehensively outlines the establishment of HP ¹³C-MRI, covering multidisciplinary team collaboration, hardware prerequisites, probe preparation, hyperpolarization techniques, imaging acquisition, and data analysis. This article discusses the clinical applications of HP ¹³C-MRI across various anatomical domains, including the brain, heart, skeletal muscle, breast, liver, kidney, pancreas, and prostate. Each section highlights the specific applications and findings pertinent to these regions, emphasizing the potential versatility of HP ¹³C-MRI in diverse clinical contexts. This review serves as a comprehensive update, bridging technical aspects with clinical applications and offering insights into the ongoing advancements in HP ¹³C-MRI.

• Investigative Magnetic Resonance Imaging
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• 제20권1호
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• pp.53-60
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• 2016

Purpose: To develop a technique for quantifying the $^{13}C$-metabolites by performing frequency-selective hyperpolarized $^{13}C$ magnetic resonance spectroscopy (MRS) in vitro which combines simple spectrally-selective excitation with spectrally interleaved acquisition. Methods: Numerical simulations were performed with varying noise level and $K_p$ values to compare the quantification accuracies of the proposed and the conventional methods. For in vitro experiments, a spectrally-selective excitation scheme was enabled by narrow-band radiofrequency (RF) excitation pulse implemented into a free-induction decay chemical shift imaging (FIDCSI) sequence. Experiments with LDH / NADH enzyme mixture were performed to validate the effectiveness of the proposed acquisition method. Also, a modified two-site exchange model was formulated for metabolism kinetics quantification with the proposed method. Results: From the simulation results, significant increase of the lactate peak signal to noise ratio (PSNR) was observed. Also, the quantified $K_p$ value from the dynamic curves were more accurate in the case of the proposed acquisition method compared to the conventional non-selective excitation scheme. In vitro experiment results were in good agreement with the simulation results, also displaying increased PSNR for lactate. Fitting results using the modified two-site exchange model also showed expected results in agreement with the simulations. Conclusion: A method for accurate quantification of hyperpolarized pyruvate and the downstream product focused on in vitro experiment was described. By using a narrow-band RF excitation pulse with alternating acquisition, different resonances were selectively excited with a different flip angle for increased PSNR while the hyperpolarized magnetization of the substrate can be minimally perturbed with a low flip angle. Baseline signals from neighboring resonances can be effectively suppressed to accurately quantify the metabolism kinetics.

• 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.

• Journal of Microbiology and Biotechnology
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• 제13권4호
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• pp.628-631
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• 2003

The recombinant alanine dehydrogenase (ADH) from E. coli containing Thermus caldophilus ADH was purified to homogeneity from a cell-free extract. The enzyme was purified 38-fold with a yield of 68% from the starting cell-free extract. The purified enzyme gave a single band in polyacrylamide gel electrophoresis, and its molecular weight was estimated to be 45 kDa. The pH optimum was 8.0 for reductive amination of pyruvate and 12.0 for oxidative deamination of L-alanine. The enzyme was stable up to $70^{\circ}C$. The activity of the enzyme was inhibited by 1 mM $Zn^{2+}$, 20% hexane, and 20% $CHCl_3$. However, 10 mM $Mg^{2+}$ and 40% propanol had no effect on the enzyme activity. The Michaelis constants ($K_m$) for the substrates were $50\;\mu\textrm{M}$ for NADH, 0.2 mM for pyruvate, 39.4 mM for $NH_4+$, 2.6 mM for L-alanine, and 1.8 mM for $NAD^+$.

• 생명과학회지
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• 제20권2호
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• pp.260-268
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• 2010

가물치(Channa argus) 조직의 젖산탈수소효소 동위효소(EC 1.1.1.27, lactate dehydrogenase, LDH)를 정제하고 생화학적, 면역학적 및 역학적 방법으로 특성을 연구하였다. LDH 활성은 골격근이 380.4 units로 가장 높고 심장 13.4, 눈 3.5, 뇌 조직 5.4 units이었으며, 심장의 CS 활성은 20.7 unit로 가장 높고, LDH/CS는 골격근 172.9, 심장 0.6, 눈 0.32, 뇌 0.47이고, 단백질 양은 골격근 14.7 mg/g이며, 특이활성(units/mg)은 골격근 25.88, 심장 0.79, 눈 0.31, 뇌 1.38 units/mg이었으므로 골격근은 혐기적이고, 심장은 호기적이었다. LDH $A_4$, $B_4$, eye-specific $C_4$에 대한 항혈청을 사용한 Western blot, 면역침강반응 및 native-polyacrylamide gel electrophoresis에 의해 $A_4$, $A_3B$, $A_2B_2$, $AB_3$ 및 $B_4$가 모든 조직에서 확인되었고, 눈 조직에서 $C_4$와 $AC_3$, $A_2C_2$, $AC_3$, 뇌 조직에서 $A_3C$도 확인되었다. LDH $A_4$, $A_3B$, $A_2B_2$, $AB_3$, $B_4$, eye-specific $C_4$ 동위효소는 affinity chromatography와 Preparative PAGE Cell에 의해 정제되었다. LDH $A_4$ 동위효소는 $NAD^+$ 유입 후 정제되었고, eye-specific $C_4$는 $A_4$에 이어 용출되기 시작하였으며 $B_4$는 buffer 유입 후 용출되었다. 정제한 결과 $A_4$는 $B_4$ 및 eye-specific $C_4$와 분자구조의 일부가 유사하였지만 $B_4$와 $C_4$는 서로 다른 것으로 나타났으므로, 하부단위체 A는 보존적이고, 하부단위체 B는 A보다 더 빠르게 진화된 것으로 보인다. 피루브산 10 mM에서 $A_4$ 동위효소 39.98%, $A_2B_2$ 21.28%, $B_4$ 19.67% 및 eye-specific $C_4$ 16.87%의 활성이 남아있었고, 피루브산에 대한 $Km^{PYR}$은 $A_4$ 0.17 mM, $B_4$ 0.27 mM, eye-specific $C_4$ 0.133 mM였다. $A_4$, $B_4$, eye-specific $C_4$, $A_2B_2$, $A_3B$ 및 $AB_3$의 최적 pH는 각각 pH 6.50, pH 8.5, pH 5.5, pH 6.0-6.5, 5.0 및 pH 7.5였고, 동질사량체 $A_4$와 이질사량체 동위효소들은 넓은 pH 영역에서 안정하였다. 특히 골격근은 LDH 활성이 크므로 활동성이 크며, 눈조직에서 피루브산 친화력이 강한 eye-specific $C_4$에 의해 피루브산 대사가 빠르게 일어나고, 이어서 $A_4$에 의해 젖산이 산화되어지는 것으로 사료되므로, 종의 생태환경 및 먹이 획득 양식에 따라 LDH-C 발현, 기질에 대한 친화도 및 대사 시간이 다른 것으로 사료된다.

• 한국식품영양과학회지
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• 제37권9호
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• pp.1174-1181
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• 2008

열처리 온도를 달리한 마늘의 이화학적 성분 및 항산화활성을 비교하고자 통마늘을 지퍼백에 $400{\pm}20\;g$씩 넣어 60, 70, 80 및 $90^{\circ}C$로 조절한 incubate에 숙성하면서 1, 3, 6일에 시료를 취하여 이화학적 분석을 실시하였으며 각 열수 및 에탄올 추출물의 DPPH에 대한 전자공여능과 환원력을 통하여 항산화활성을 분석하였다. 열처리 온도를 달리하여 숙성한 마늘의 L, a 및 b값 모두 숙성온도가 높을수록 숙성기간이 경과할수록 유의적으로 낮아졌는데, $60^{\circ}C$에서 6일간 숙성한 시료의 b값은 $22.16{\pm}0.76$이었으나 $90^{\circ}C$ 숙성 시료는 $1.35{\pm}0.21$로 급격히 낮아졌다. 전단가의 경우 숙성 초기에는 $90^{\circ}C$ 숙성 시료가 $85.70{\pm}1.44\;cm/kg^2$로 가장 낮았으나 숙성기간이 경과할수록 경도가 증가하여 숙성 6일에는 $411.30{\pm}13.90\;cm/kg^2$으로 증가하였다. 마늘의 pH는 숙성온도가 높을수록 숙성 기간이 경과할수록 산성화되었는데 6일 숙성 후 $60^{\circ}C$와 $90^{\circ}C$ 시료의 pH는 각각 6.12와 3.90으로 큰 차이를 보였다. 총페놀과 플라보노이드의 함량은 숙성온도가 높을수록 숙성 기간이 경과할수록 증가하여 숙성 6일 시료의 경우 $60^{\circ}C$ 숙성 시료에 비하여 $90^{\circ}C$ 숙성 시료에서 총 페놀은 약 3.5배, 플라보노이드는 약 9.1배 더 높은 함량이었다. Total pyruvate의 경우 숙성 온도에 따라 증감의 경향이 서로 상이하여 $60^{\circ}C$와 $70^{\circ}C$ 숙성 시료에서는 숙성기간 1일과 6일을 비교할 때 더 감소하였으나 $80^{\circ}C$와 $90^{\circ}C$ 숙성 시료의 경우 숙성 6일에 오히려 그 값이 유의적으로 증가되었다. DPPH에 대한 항산화활성은 마늘의 숙성 온도가 높을수록, 숙성 기간이 경과할수록, 농도가 진할수록 증가하여 $90^{\circ}C$, 6일 숙성 시료의 항산화활성은 $87.48{\pm}0.20%$로 타 시료의 $43.61{\sim}55.76%$ 보다 월등히 높았고 환원력도 이와 동일한 경향이었다. 이처럼 항산화 활성이 증가하는 것은 마늘의 숙성온도가 높을수록 숙성 기간이 경과할수록 갈변화 물질의 생성이 많아지고, 이에 따라 총페놀과 플라보노이드 함량이 증가하는 것으로 볼 때 마늘 중의 항산화성 갈변 물질의 증가에 기인하는 결과로 판단된다.

• 대한유전성대사질환학회지
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• 제17권2호
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• pp.69-76
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• 2017

Citrin 결핍증은 요소회로 이상 질환 중 하나로, 7q21.3에 위치한 SLC25A13 유전자에 돌연변이로 발생하는 상염색체 열성 유전질환이다. 세 가지 표현형 중에 신생아 간내 담즙 정체형, 제 2형 시트룰린혈증은 잘 알려졌지만, 성장부진과 이상지질혈증형은 최근에 밝혀지고 있는 표현형으로 아직 우리나라에 보고된 적이 없다. 성장부진과 이상지질혈증형에서는 경미할 수는 있으나 식욕감소, 피곤함, 성장부진, 저혈당, 시트룰린 상승, 이상지질혈증, 젖산염/피루브산염 상승과 같은 이상이 관찰될 수 있다. 또한 저혈당으로 내원하였을 때 일반적인 검사로는 원인 규명이 어려울 수 있다. 저자들은 생후 30개월에 반복적인 저혈당으로 내원하여 소변 유기산 분석, 호르몬 검사와 같은 대사 이상 검사에서 명확한 특정 진단명이 의심되지 않아, 생후 31개월에 targeted exome sequencing을 통해 복합이형접합 SLC25A13 유전자 돌연변이[c.852_855del (p.Met285Profs*2), c.1177+1G>A]를 발견하여 성장부진과 이상지질혈증으로 발현한 citrin 결핍증을 우리나라에서 최초로 진단하여 보고하는 바이다.

• Natural Product Sciences
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• 제13권2호
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• pp.105-109
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• 2007

Alcoholic extract of Terminalia arjuna [TA] was evaluated far its hepatoprotective activity against carbon tetrachloride (CCl$_4$)-induced hepatic damage in rats. The hepatoprotective activity of TA was evaluated by measuring levels of serum marker enzymes like serum glutamate oxaloacetate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT), serum alkaline phosphatase (SALP). The serum levels of total proteins(TP), total albumins (TAL) and bilirubin (BILN) were also estimated. The histological studies were also carried out to support the above parameters. Silymarin (SM) was used as standard drug. Administration of TA (250 and 500 mg/kg/po) markedly prevented CCl$_4$-induced elevation of levels of SGOT, SGPT, SALP, TP, TAL and BILN. These biochemical observations were supplemented by histopathological examination of liver sections. Alcoholic extract of TA also shown significant in-vitro free radical scavenging activity against 1, 1-diphenyl-2-picryl hydrazyl (DPPH) and nitric oxide (NO) radicals. Thus, the present study provides a scientific rationale for the traditional use of this plant in the management of liver diseases.