Purpose: Several radioisotope-labeled thymidine derivatives such as $[^{11}C]$thymidine was developed to demonstrate cell proliferation in tumor. But it is difficult to track metabolism with $[^{11}C]$thymidine due to rapid in vivo degradation and its short physical half-life. 3'-$[^{18}F]$fluoro-3'-deoxythymidine ($[^{18}F]$FLT) was reported to have the longer half life of fluorine-18 and the lack of metabolic degradation in vivo. Here, we described the synthesis of the 3'-$[^{18}F]$fluoro-3'-deoxythymidine ($[^{18}F]$FLT) and compared with $([^{18}F]FET)\;and\;([^{18}F]FDG)$ in cultured 9L cell and obtained the biodistribution and PET image in 9L tumor hearing rats. Material and Methods: For the synthesis of $[^{18}F]$FLT, 3-N-tert-butoxycarbonyl-(5'-O-(4,4'-dimet hoxytriphenylmethyl)-2'-deoxy-3'-O-(4-nitrobenzenesulfonyl)-${\beta}$-D-threopentofuranosyl)thymine was used as a FLT precursor, on which the tert-butyloxycarbonyl group was introduced to protect N3-position and nitrobenzenesulfonyl group. Radiolabeling of nosyl substitued precursor with $^{18}F$ was performed in acetonitrile at $120^{\circ}C$ and deproteced with 0.5 N HCI. The cell uptake was measured in cultured 9L glioma cell. The biodistribution was evaluated in 9L tumor bearing rats after intravenous injection at 10 min, 30 min, 60 min and 120 min and obtained PET image 60 minutes after injection. Results: The radiochemical yield was about 20-30% and radiochemical purity was more than 95% after HPLC purification. Cellular uptake of $[^{18}F]$FLT was increased as time elapsed. At 120 min post-injection, the ratios of tumor/blood, tumor/muscle and tumor/brain were $1.61{\pm}0.34,\;1.70{\pm}0.30\;and\;9.33{\pm}2.22$, respectively. The 9L tumor was well visualized at 60 min post injection in PET image. Conclusion: The uptake of $[^{18}F]$FLT in tumor was higher than in normal brain and PET image of $[^{18}F]$FLT was acceptable. These results suggest the possibility of $[^{18}F]$FLT at an imaging agent for brain tumor.
Purpose: The HSV1-tk reporter gene system is the most widely used system because of its advantage that direct monitoring is possible without the introduction of a separate reporter gene in case of HSV1-tk suicide gene therapy. In this study, we investigate the usefulness of the reporter probe (substrate), $9-(4-[^{18}F]Fluoro-3-hydroxymethylbutyl)$guanine ($[^{18}F]FHBG$) for non-invasive reporter gene imaging using PET in HSV1-tk expressing hepatoma model. Materials and Methods: Radiolabeled FHBG was prepared in 8 steps from a commercially available triester. The labeling reaction was carried out by NCA nucleophilic substitution with $K[^{18}F]/K2.2.2.$ in acetonitrile using N2-monomethoxytrityl-9-14-(tosyl)-3-monomethoxytritylmethylbutyl]guanine as a precursor, followed by deprotection with 1 N HCl. Preliminary biological properties of the probe were evaluated with MCA cells and MCA-tk cells transduced with HSV1-tk reporter gene. In vitro uptake and release-out studies of $[^{18}F]FHBG$ were performed, and was analyzed correlation between $[^{18}F]FHBG$ uptake ratio according to increasing numeric count of MCA-tk cells and degree of gene expression. MicroPET scan image was obtained with MCA and MCA-tk tumor bearing Balb/c-nude mouse model. Results: $[^{18}F]FHBG$ was purified by reverse phase semi-HPLC system and collected at around 16-18 min. Radiothemical yield was about 20-25%) (corrected for decay), radiochemical purity was >95% and specific activity was around >55.5 $GBq/{\mu}\;mol$. Specific accumulation of $[^{18}F]FHBG$ was observed in HSV1-tk gene transduced MCA-tk cells but not in MCA cells, and consecutive 1 hour release-out results showed more than 86% of uptaked $[^{18}F]FHBG$ was retained inside of cells. The uptake of $[^{18}F]FHBG$ was showed a highly significant linear correlation ($R^2=0.995$) with increasing percentage of MCA-tk numeric cell count. In microPET scan images, remarkable difference of accumulation was observed for the two type of tumors. Conclusion: $[^{18}F]FHBG$ appears to be a useful as non-invasive PET imaging substrate in HSV1-tk expressing hepatoma model.
Proceedings of the Korean Society for Food Science of Animal Resources Conference
/
1998.05a
/
pp.37-54
/
1998
This study was conducted to investigate the effects of mugwort extracts on the blood ethanol concentration, liver function and low level of cadmuim(Cd) in rats. The effects of mugwort extracts on the blood ethanol concentration was studied in Sprague-Dawley rats (10 weeks old) administered p.o. with 25% ethanol (5g/1kg body weight) and then injected with mugwort extracts (at the 2% levels of daily feed consumption compared with the concentration of catechins level in mugwort extracts) in caudal vein. SD rats were divided into five groups : control group (CON-E, only ethanol and 0.85% saline sol'n treated instead of each extracts), water extracts of mugwort treated to the control (MDW-E), ethanol extracts of mugwort treated to the control (POH-E). And then rat plasma of each time (0hr, 1hr, 2hr, 3hr) was investigated ethanol concentration by gas chromatography. Another rats were measured at the time of 0 and 5hr for the test of GOD(Glutamic Oxaloacetic Transaminase) and GPT(Glutamic Pyruvic Transaminase). Components of each extracts were analyzed by using high performance liquid chromatography. The effects of mugwort extracts on the liver function were studied in culture of rat hepatocyte composed of three groups : Control group and two groups treated with each extracts (1% & 2% MDW, 1% & 2% MOH). Condition of rat hepatocytes cultured for 36hr at $37^{\circ}C$(5% $CO_2$ incubator), number of cells, GOT and GPT activity were investigated. The results obtained were summarized as follows ; 1. Catechins level of mugwort extracts was $8{\sim}10mg/100g(MDW)$, $3{\sim}4mg/100g(MOH)$ 2. The contents of (-)-Epigallocatechin was high in MDW 3. The effects of mugwort extracts on the blood ethanol concentration were as follows; 1) The order in ethanol degradation efficiency was MDW-E > MOH-E > CON-E. 2) Ethanol concentration significantly decreased (p<0.05) in MDW-E and MOH-E. 4. The effects of mugwort extracts on the liver function were as follows; (rat hepatocytes cultured for 36hr at $37^{\circ}C$) 1) Cells condition of MDW-L was better than other groups. 2) The order in number of cells (rat hepatocytes) was 2% MDW-L >1% MDW-L >1% MOH-L > Con-L > 2% MOH-L 5. Cd treatment increased concentrations of hepatic GSH level, and decreased GOT activity in plasma. Therefore, this results suggest that the effects of mugwort extracts may an important rols in degradation ethanol and recovery liver function in body. Also, Mugwort extracts may modify the toxicities of Cd in Cd-treated rats and play an important roles in preventing the liver from various toxicants including Cd in Cd treated rats.
Kim, Ju-Hye;Kim, Jong-Hwan;Kim, Dae-Wook;Lee, Bong-Jae;Kim, Chansub;Ihm, Yangbin;Seo, Jong-Su
The Korean Journal of Pesticide Science
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v.19
no.3
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pp.174-184
/
2015
In the present study, the metabolism of [$^{14}C$]butachlor was investigated in rice plant according to the OECD test guideline No. 501. [$^{14}C$]Butachlor was treated as granule to paddy water by application of 1.5 kg ingredient (a.i.)/ha at the 3~4 leave stage of rice plant. At 85 days after treatment (DAT), samples of panicle, foliage, and roots were taken for radioactivity analysis. Upon harvest at 126 DAT, rice plants were separated into brown rice, husk, straw, and root parts. Amounts of total radioactivity absorbed by rice plant ranged from 8.6 to 9.8% of applied radioactivity (AR). Total radioactive residues (TRRs) of rice plant at 126 DAT was the highest as 4.0421 mg/kg (7.3% AR) in the straw followed by 1.4595 mg/kg (2.4% AR) in the root, 0.7257 mg/kg (0.1% AR) in the husk. The lowest level recording 0.1020 mg/kg (0.1% AR) was found in brown rice. Each part was extracted with various solvents and solvent/water mixtures. Greater than 70% of TRRs was readily extractable from foliage, panicle, husk and straw. Only 34.0% of the brown rice and 43% of root based on TRRs were extractable showing that the residues were completely assimilated in the plant tissue. The level of non-extractable radioactivity was ranged from 26.2 to 66.0% of TRRs. From this study, five tentative major metabolites (M1, M2, M3, M4 and M5) were observed in rice extracts. Among the metabolites, 2,6-diethylaniline assigned as M4 was identified in rice plant by comparing to retention time of reference standard. Un-metabolized butachlor was not detected in any fractions. In soil extracts, N-(butoxymethyl)-N-(2,6-diethyl phenyl)acetamide, 2,6-diethylaniline, M2, M3 and M5 were observed. And the concentration of butachlor was low level (ca. 0.03 mg/kg).
To provide a basis for the variation of fruit bodies of winter mushroom (Flammulina velutipes), the organic acid composition of its fruit bodies was investigated with several varieties of winter mushroom indifferent temperature and storage period. In the fruit bodies of winter mushroom, a total of 10 organic acids including acetic acid, butyric acid, citric acid, fumaric acid, DL-isocitric acid, L(+)lactic acid, D-malic acid, propionic acid, succinic acid, and D-tartaric acid were detected. In the most of the winter mushroom, acetic acid was the main organic acid component and fumaric acid was the least included component. Acetic acid, which is a mono-carboxyl group of organic acid, is contained in different levels according to different varieties and different storage temperature. Butyric acid is extremely variable in its quantity, depending on variety and different storage temperature. In contrast, fumaric acid, which is a dicarboxyl group of organic acid, decreased in its quantity during storage with 1.5 mg/g. Especially, ASI 4149, 4166 varieties tend to differ in their quantity. Besides, malic acid is extremely variable in its quantity according to variety and storage temperature. Citric acid, a tri-carboxyl group of organic acid, increased in its quantity according to storage period, which enables us to efficiently manage storage period. Isocitric acid is also extremely variable in its quantity according to variety, storage temperature and storage period.
Sung, Jung Sook;Song, Seok Bo;Kim, Ji Young;An, Yeon Ju;Park, Jae Eun;Choe, Myeong Eun;Chu, Ji Ho;Ha, Tae Joung;Han, Sang Ik
KOREAN JOURNAL OF CROP SCIENCE
/
v.65
no.3
/
pp.231-240
/
2020
This research was conducted to evaluate the physicochemical properties, antioxidant components, and their activities for more taking advantage of small redbean cultivars. Seed size, 100 seeds weight, and hardness on the 8 cultivars were measured. The free sugar and crude protein contents were evaluated using HPLC and protein analyzer, respectively. Amylose content, antioxidant components, and activities were analyzed by spectrophotometer. The range of 100 seeds weight and hardness were 12.55-18.81 g and 9,527.38-14,341.25 gf, respectively. Total free sugar, amylose, and crude protein were showed 22.49-31.07 mg/g, 13.53-15.67%, and 21.27-23.30%, respectively. The cultivar 'Hongeon' was higher antioxidant component and activity more than others. In clustering the cultivars based on the results, the tree showed four major clades. The 'Huinnarae' group was high in total free sugar and amylose content. The 'Hongeon' group were high in 100 seeds weight, antioxidant component. and activity, while amylose content was lower than that in the other groups. The results of the cultivars can be utilized for research of functional materials. The findings of this study will provide valuable information for expansion of functional food industry related on small redbean.
Cervical cancer is one of the leading causes of female death. Viral oncoproteins E6 and E7 are selectively retained and expressed in carcinoma cells infected with HPV (Human papillomavirus) type 16. The HPV is cooperated in immotalization and transformation of primary keratinocyte. E6 and E7 oncoproteins interfere the functions of tumor suppressor proteins p53 and retinoblasoma protein (pRb), respectively. Among a lots of natural products, Artemisia scoparia Waldstein et Kitamura has inhibitory effects on the binding between E6 oncoprotein and tumor suppressor p53, or the binding between E6 and E6 associated protein (E6AP), an E3 ubiquitin-protein ligase. HPV oncoprotein inhibitors from Artemisia scoparia W. were isolated by solvent partition and column chromatography (Silica gel, RP-18) and the inhibitory compounds were finally purified by HPLC using an ELISA screening system based on the binding between E6 and E6AP. The aim of this study is to identify the structure of inhibitory compounds and to investigate whether these compounds have inhibitory effects on the functions of E6 oncoprotein. We investigated whether 3,5-di-O-caffeoylquinic acid (DCQA) extracted from Artemisia scoparia W. Could inhibit the function of E6 oncoprutein. DCQA inhibited the in vitro binding of E6 and E6AP which are essential for the binding and degradation of the tumor suppressor p53 and also inhibited the proliferation of human cervical cancer cell lines (SiHa and CaSKi) in a dose response manner. These results suggest that DCQA inhibited the function of E6 oncoprotein, suggesting that it can be used as a potential drug for the treatment of cervical cancers infected with HPV.
Jo, Ji-Song;Nguyen, Do Quynh Anh;Yun, Jun-Ki;Kim, Yu-Na;Kim, You-Geun;Kim, Sung-Bae;Seo, Yang-Gon;Lee, Byung-Hak;Kang, Moon-Kook;Kim, Chang-Joon
Microbiology and Biotechnology Letters
/
v.37
no.3
/
pp.231-237
/
2009
This paper aimed to develop a solvent extraction and purification process to recover high-purified ${\beta}$-carotene from recombinant Escherichia coli. Cells harvested from the culture broth were treated through numerous steps: dehydration, solvent extraction, crystal formation and separation. To optimize the extracting condition, experiments were carried out to investigate the effect of cell disruption, temperature, organic solvents, solvent-biomass ratio on the yield of ${\beta}$-carotene extracted from cells. The result indicated that no significant differences of extraction yield were observed from cells with or without step of cell disruption. Among different extracting solvents, the highest extraction yield of ${\beta}$-carotene, 30.3 mg-${\beta}$-carotene/g-dry cells, was obtained with isobutyl acetate at solvent-biomass ratio 25 mL/g-dry cells at $50^{\circ}C$. Notably, in case of acetone, the extraction yield was quite low when using acetone itself, but increased almost up to the highest value when combining this solvent and olive oil. The purity of ${\beta}$-carotene crystals obtained from crystallization and separation was 89%. The purity degree was further improved up to 98.5% by treating crude crystals with additional ethanol washing.
Kwon, Soon Woo;Ko, Hyun Ju;Bae, Jun Tae;Kim, Jin Hwa;Lee, Geun Soo;Pyo, Hyeong Bae
Journal of the Society of Cosmetic Scientists of Korea
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v.42
no.1
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pp.75-85
/
2016
Pectin, a naturally occurring polysaccharide, has in recent years attracted considerable attention. Its benefits are increasingly appreciated by scientists and consumers due to its safety and usefulness. The chemistry and gel-forming characteristics of pectin have enabled to be used in pharmaceutical industry, health promotion and treatment. Yet, it has been rarely used in cosmetics because of its incompatibility with many cosmetic ingredients, including alcohols, and unstable viscosity of pectin gels under various pH and salt conditions. However, low-molecular-weight pectin oligomers have excellent biological activities, and depolymerization of pectin to produce cosmetic ingredients would be very useful. In this study, we attempted the development of cosmetic ingredients using pectin with an excellent effect on human skin. We developed a bio-conversion process that uses enzymatic hydrolysis to produce pectin hydrolysates containing mainly low-molecular-weight pectin oligomers. Gel permeation chromatography was used to determined the ratio of hydrolysis. The molecular weight of the pectin hydrolysates obtained varied between 200 and 2,700 Da. The two newly developed low-molecular-weight pectin hydrolysates, LMPH A and B, had higher anti-oxidative activities than pectin or D-galacturonic. Exposure to UVB radiation induces apoptotic cell death in epidermal cells. Annexin V binding and propidium iodide uptake were measured by flow cytometry to evaluate UVB-induced cell death in HaCaT cells. Both LMPH A and B reduced UVB-induced cell death and increased cell proliferation by 22% and 30% at 0.5% concentration respectively, while pectin had no significant activity. In conclusion, this study suggests that the newly developed low-molecular-weight pectin hydrolysates can be used as safe and biologically active cosmetic ingredients.
Won, Beom Young;Shin, Ki Young;Ha, Hyun Jee;Wee, Ji-Hyang;Yun, Yeo Sang;Kim, Ye Ri;Park, Yong Jin;Jung, Kyoung Ok;Sung, Hea Mi;Lee, Hyung Gun
Journal of the Korean Society of Food Science and Nutrition
/
v.45
no.1
/
pp.1-11
/
2016
This study was conducted to investigate effect of the acetylcholinesterase inhibitor activity, the protective effect of the extract on SH-SY5Y cell death by $H_2O_2$, the memory improvement from scopolamine-induced rat. Moreover, the antioxidant activity of isorhamnetin from the dropwort (Oenanthe javanica) was investigated. Acetylcholinesterase inhibitor activity was highest (28.59%) in Hwasun O. javanica extract (H-OJE). H-OJE and Naju O. javanica extract (N-OJE) were not significantly different. SH-SY5Y cell death deceased to 37.23% and 36.68% for H-OJE and N-OJE, respectively, following treatment with the extracts. O. javanica extracts showed a protective effect against $H_2O_2$-induced neurotoxicity. Treatment with O. javanica extracts slightly improved scopolamine-induced (1 mg/kg, i.p.) memory impairment in rats. H-OJE contained the highest total phenolic and flavonoid contents of 117 mg/g and 30 mg gallic acid equivalents/g, respectively, and had a DPPH radical scavenging activity ($SC_{50}$) of $113.8{\mu}g/mL$ and ABTS radical scavenging activity of $48.2{\mu}g/mL$, which was higher than the other extracts. The thiobarbituric acid reactive substances value was highest (50.2%) in H-OJE. Antioxidant activity differed significantly among dropwort extracts. Isorhamnetin was known as one of the flavonoid and for having neuroprotective effect. So we analyzed acid-hydrolyzed O. javanica extract HPLC. The results were that peak at 14 min and spectrum of the extracts was consistent with standard solution. The results of LC/MS/MS analysis were that the extract and standard solution were confirmed total ion chromatogram at identical time, precursor ion was 317 $[M-H]^+$ m/z, product ion was 302 $[M-H]^+$ m/z. Overall, the results showed that the dropwort extract led to memory improvement and had antioxidant activity. Based on these finding, further research to investigate the production of ethanol extract of dropwort as a processed food is warranted.
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