• KOREAN JOURNAL OF CROP SCIENCE
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• v.37 no.5
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• pp.468-475
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• 1992

In order to understand more clearly the integration between N-assmilation and C-metabolism in relation to N fertilization, a pot experiment with 5 different level of N fertilization(0, 5, 10, 25, 50 mM NO$_3$$_{[-10]}$ ) was conducted in Manchester, U.K. The peas (Pisum sativum L., cv. Early Onward) were sown in vermiculate (5 cm depth) and cultivated for 6 days under temperature controlled dark room conditions ($25^{\circ}C$). The plants received frequent irrigation with a nutrient solution: it was fertilized every 2 days, 3 times a day at 10h, 13h, 16h respectively. Elevated NO$_3$$^{[-10]}$ concentration, the activity levels of NR, NiR, total GS(crude extract), GS$_2$(plastid) in both root and shoot were increased and reached the peak in 5～25 mM, except NiR specific activity which increased its activity continually until 50 mM NO$_3$$^{[-10]}$ treatment. Total activities of GS (crude extract) in both root and shoot became higher than those of GS$_2$(Plastid), and the activity ratios of total GS in the crude extract and GS$_2$ in the plastids were 3.0 to 4.3 in root, but 3.2 to 10.6 in shoot. It was concluded that the reductants and A TP from OPPP itself should be enough to achieve the high rate of NR, NiR, GS$_1$, GS$_2$ in plant root and shoot for reduction or assimilation of nitrogen, but these enzyme activities might be inhibited by an excess of NO$_3$$^{[-10]}$ influx over the reduction capacity.

• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.6 no.1
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• pp.81-97
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• 2000

Objectives: This experimental study was carried out to evaluate the effects of aqueous and methanol extracts of Hedyotis diffusa which has long been used for cancer treatment in oriental medicines on the induction of apoptotic cell death in human lymphoid leukemia cell line, HL-60. Methods: Cells were treated with various concentrations (200 to $0.4{\mu}g$) and periods (6 to 30 hr) of $H_2O$ and methanol extracts of Hedyotis diffusa. Then, cells were tested for viability by MTT assay. Cells wrere treated with $200{\mu}g/ml$ of methanol extract fork various periods. Genomic DNA was isolated, separated, on 1.5% agarose gels, stained with ethidium bromide and visualized under UV light. Cells were treated with $200{\mu}g/ml$ of each extract for 16 hr. Then, cells were treated with Hoechst dye 33342 and observed by fluorescence microscopy. Cells were treated with various doses of each for 12 hr and $100{\mu}g/ml$ of methanol extract for various periods. Lysate from the cells used to measure the activity of Caspase-1 and-3 proteases by using fluorogenic peptide substrates including acetyl-YVAD-AMC and acetyl-DEVD-AMC, respectively. Cells were treated with $200{\mu}g/ml$ of each extract for various periods. Cell lysates were immunoprecipated with anti-JNKl antibodies. The immune complex was reacted with $32^p-ATP$ and c-Jun as a substrate. The phosphotransferase activity of JNKI was measured by using PhosphoImage analyzer (Fuji Co., Japan). Nuclear extracts were isolated and incubated with oligonucleotide probe of $NF-{\kappa}B$. Transcriptional activation of ${\kappa}B$ was measured by using EMSA and visualized by PhosphoImage analyzer (Fuji Co, Japan). Cell lysates were prepared and analyzed by Western blotting with anti-Bc12 antibodies and anti-Bax antibodies. Cells were pretreated with various doses of methanol extract for 2 hr. Then, the extract was removed by centrifugation. Cells were resuspended with RPMI-1640 media containing 0.3% agarose, 10% FBS, overlayred onto bottom layer agarose and incubated at $CO_2$ incubator for 6 days. The number of colony was counted under light microscopy ($\time100$). Results: The death of HL-60 cells was markedly induced by the addition of methanol extract of Hedyotis diffusa in a dose and time-dependent manners. The apoptotic characteristic ladder pattern of DNA strand break was observed in death of HL-60 cells. In addition, it was shown nucleus chromatin condensation and fragmentation under Hoechst staining. Therefore, Hedyotis diffusa extract-induced death of HL-60 cells is mediated by apoptotic signaling processes. The activity of Caspase 3-like proteases remained in a basal level in HL-60 cells treated with aqueous extract of Hedyotis diffusa. However, it was markedly increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. In addition, the phosphotransferase activity of JNKl was increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. Furthermore, the activation of transcriptional activator, $NF-{\kappa}B$ was markedly induced by methanol extract of Hedyotis diffusa. Anti-apoptotic Bc12 was cleaved into 23Kda fragment by treatment of methanol extract of Hedyotis diffusa. However, expression of proapoptotic Bax protein was increased by treatment of methanol extract of Hedyotis diffusa in a time-dependent manner. Furthermore, methanol extract markedly inhibited the colony forming efficiency of HL-60 cells in semisolid agar culture. Conclusions: Above results suggest that methanol extract of Hedyotis diffusa induces the apoptotic death of human leukemic HL-60 cells via activations of Caspase-3 proteases, JNKI, transcriptional activator $NF-{\kappa}B$, In addition, our results also suggest that methanol extract of Hedyotis diffusa reduces the malignant potential of HL-60 cells via down regulation of colony forming effciency through cleavage of Bc12 as well as induction of Bax.

• The Korean Journal of Nuclear Medicine
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• v.38 no.2
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• pp.180-189
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• 2004

Although the outcome of cancer patients after cytotoxic chemotherapy is related diverse mechanisms, multidrug resistance (MDR) for chemotherapeutic drugs due to cellular P-glycoprotein (Pgp) or multidrug-resistance associated protein (MRP) is most important factor in the chemotherapy failure to cancer. A large number of pharmacologic compounds, including verapamil, quinidine, tamoxifen, cyclosporin A and quinolone derivatives have been reported to overcome MDR. Single photon emission computed tomography (SPECT) and positron emission tomography (PET) are available for the detection of Pgp and MRP-mediated transporter. $^{99m}Tc$-MIBI and other $^{99m}Tc$-radiopharmaceuticals are substrates for Pgp and MRP, and have been used in clinical studies for tumor imaging, and to visualize blockade of PgP-mediated transport after modulation of Pgp pump. Colchicine, verapamil and daunorubicin labeled with $^{11}C$ have been evaluated for the quantification of Pgp-mediated transport with PET in vivo and reported to be feasible substrates with which to image Pgp function in tumors. Leukotrienes are specific substrates for MRP and $N-[^{11}C]acetyl-leukotriene$ E4 provides an opportunity to study MRP function non-invasively in vivo. SPECT and PET pharmaceuticals have successfully used to evaluate pharmacologic effects of MDR modulators. Imaging of MDR and reversal of MDR with bioluminescence in a living animal is also evaluated for future clinical trial. We have described recent advances in molecular imaging of MDR and reviewed recent publications regarding feasibility of SPECT and PET imaging to study the functionality of MDR transporters in vivo.

• Proceedings of the SOHE Conference
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• 1997.12a
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• pp.39-43
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• 1997

Toha-jeod was manufactured by seven methods ; low salt group (L:15% sodium chloride), high salt g group (H:23% sodium chloride), 50% conventional soybean sauce group (S), low salt group containing 2% w wheat bran (W2%-L), high saIt group containing 2% wheat bran (W2%-H),high salt group containing 2% wheat bran (W2%-H), high salt group containing 4% wheat bran (W4%-H). After these seven groups were refrigerated at $4{\pm}1^{\circ}C$, they were sampled at intervals of three months and analyzed functional components. The free amino acid in Toha-jeod which are omitine, glutamic acid, leucine, alanine, lysine and valine increased gradually up to six months of fermentation and decreased by nine months. Conventional soybean sauce group increased continuously during the fermentation process. Hypoxanthine was altered almost among other nueletides. ATP was not detected, IMP and inosine had disapapted after the six months fermentation. Polyene fatty acids and n-3 fatty acids were decreased and s saturated fatty acids were not altered in the group containing wheat bran during fermentation. In the Hunter values, the group containing wheat bran and high salt group showed lower level than the group n not containing wheat bran and low salt group. Redness indicating the value of Toha-jeod increased as Toha-jeod was fermentated. Low salt group and conventional soybean sauce group were superior to other groups in the extent of redness. As the fermentation of Toha-jeod progressed for a long time, molecular weight distribution tended to become less molecular and the formation of chitin oligosaccharides was increased significantly. After nine months of fermentation, 24.75% chitin oligosaccharides [($GlcNAd_4$ ~ ($GlcNAd_8$, M.W. 823~1789] were created in the high salt group containing 2% wheat bran. [($GlcNAd_6$. M.W. 1236J , that is NACOS-6, which was reported as an antitumor activity material, was present in 4.01~4.37% of total Toha chitin content. 66.30% chitin oligosaccharides were created in conventional soybean sauce.

• Journal of Ginseng Research
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• v.47 no.3
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• pp.408-419
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• 2023

Background: Ginsenoside compound K (CK), the main active metabolite in Panax ginseng, has shown good safety and bioavailability in clinical trials and exerts neuroprotective effects in cerebral ischemic stroke. However, its potential role in the prevention of cerebral ischemia/reperfusion (I/R) injury remains unclear. Our study aimed to investigate the molecular mechanism of ginsenoside CK against cerebral I/R injury. Methods: We used a combination of in vitro and in vivo models, including oxygen and glucose deprivation/reperfusion induced PC12 cell model and middle cerebral artery occlusion/reperfusion induced rat model, to mimic I/R injury. Intracellular oxygen consumption and extracellular acidification rate were analyzed by Seahorse multifunctional energy metabolism system; ATP production was detected by luciferase method. The number and size of mitochondria were analyzed by transmission electron microscopy and MitoTracker probe combined with confocal laser microscopy. The potential mechanisms of ginsenoside CK on mitochondrial dynamics and bioenergy were evaluated by RNA interference, pharmacological antagonism combined with co-immunoprecipitation analysis and phenotypic analysis. Results: Ginsenoside CK pretreatment could attenuate mitochondrial translocation of DRP1, mitophagy, mitochondrial apoptosis, and neuronal bioenergy imbalance against cerebral I/R injury in both in vitro and in vivo models. Our data also confirmed that ginsenoside CK administration could reduce the binding affinity of Mul1 and Mfn2 to inhibit the ubiquitination and degradation of Mfn2, thereby elevating the protein level of Mfn2 in cerebral I/R injury. Conclusion: These data provide evidence that ginsenoside CK may be a promising therapeutic agent against cerebral I/R injury via Mul1/Mfn2 mediated mitochondrial dynamics and bioenergy.

• Korean Journal of Soil Science and Fertilizer
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• v.2 no.1
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• pp.53-68
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• 1969

The nutriophysiological response of rice plant to root environment was investigated with eye observation of root development and rhizosphere in situation. The results may be summarized as follows: 1) The quick decomposition of organic matter, added in low yield soil, caused that the origainal organic matter content was reached very quickly, in spite of it low value. In high yield soil the reverse was seen. 2) In low yield soil root development, root activity and T/R value were very low, whereas addition of organic matter lowered them still wore. This might be contributed to gas bubbles around the root by the decomposition of organic matter. 3) Varietal difference in the response to root environment was clear. Suwon 82 was more susceptible to growth-inhibitine conditions on low-yield soil than Norin 25. 4) Potassium uptake was mostly hindered by organic matter, while some factors in soil hindered mostly posphorus uptake. When the organic matter was added to such soil, the effect of them resulted in multiple interaction. 5) The root activity showed a correlation coeffieient of 0.839, 0.834 and 0.948 at 1% level with the number of root, yield of aerial part and root yield, respectively. At 5% level the root-activity showed correlation-coefficient of 0.751, 0.670 and 0.769 with the uptake of the aerial part of respectively. N, P and K and a correlation-coefficient of 0.729, 0.742 and 0.815 with the uptake of the root of respectively N.P. and K. So especially for K-uptake a high correlation with the root-activity was found. 6) The nitrogen content of the roots in low-yield soil was higher than in high-yield soil, while the content in the upper part showed the reverse. It may suggest ammonium toxicity in the root. In low-yield soil Potassium and Phosphorus content was low in both the root and aerial part, and in the latter particularly in the culm and leaf sheath. 7) The content of reducing sugar, non-recuding sugar, starh and eugar, total carbohydrates in the aerial part of plants in low yield soil was higher than in high yield soil. The content of them, especially of reducing sugar in the roots was lower. It may be caused by abnormal metabolic consumption of sugar in the root. 8) Sulfur content was very high in the aerial part, especially in leaf blade of plants on low yield soil and $P_2O_5/S$ value of the leaf blade was one fifth of that in high yield soil. It suggests a possible toxic effect of sulfate ion on photophosphorization. 9) The high value of $Fe/P_2O_5$ of the aerial part of plants in low yield soil suggests the possible formation of solid $Fe/PO_4$ as a mechanical hindrance for the translocation of nutrients. 10) Translocation of nutrients in the plant was very poor and most nutrients were accumulated in the root in low yield soil. That might contributed to the lack of energy sources and mechanical hindrance. 11) The amount of roots in high yield soil, was greater than that in low yield soil. The in high-yield soil was deep, distribution of the roots whereas in the low-yield soil the root-distribution was mainly in the top-layer. Without application of Nitrogen fertilizer the roots were mainly distributed in the upper 7cm. of topsoil. With 120 kg N/ha. root were more concentrated in the layer between 7cm. and 14cm. depth. The amount of roots increased with the amount of fertilizer applied.