• Journal of Plant Biotechnology
• /
• v.36 no.1
• /
• pp.30-37
• /
• 2009

Tissue-type plasminogen activator (t-PA) is a thrombolytic agent important in fibirn clot lysis. T-PA causes fibirn-specific plasminogen activation. Six binary vectors harboring t-PA and its derivative genes were cloned and expressed in transgenic alfalfa plants. The insertion of the t-PA and its derivative genes in genomic DNA of alfalfa plants was confirmed by PCR. The presence of the t-PA and its derivative transcripts in total RNAs of the transgenic alfalfa leaves was verified by RT-PCR. ELISA experiments demonstrated that the highest level of recombinant t-PA expression was $75.1{\mu}g$/ total soluble protein (mg) in alfalfa plants. The amount of recombinant t-PA and its derivative proteins in transgenic plants was estimated to range from 9.7 to $39.5{\mu}g$/ total soluble proteins (mg). Western blot analysis of the transformed alfalfa leaves revealed bands of approximately 68-kDa recombinant t-PA and its derivative proteins. The fibrinolysis of recombinant t-PA and its derivative proteins was confirmed by a fibrin plate assay (range from 3.2 to 8.1 cm). The results presented provide information for the development of an additional production of recombinant human proteins having pharmaceutical applications using transgenic plants.

• Korean Journal of Poultry Science
• /
• v.35 no.1
• /
• pp.85-99
• /
• 2008

Avian reovirus (ARV) is a causative agent of viral arthritis/tenosynovitis, and malabsorption syndrome in broiler. The characteristics of malabsorption syndrome caused by ARV are diarrhea, poor feed conversion and stunting. Therefore, ARV infection has been recognized as one of the most important disease in the poultry industry because of economical losses. However, few study of ARV infection in broiler industry has been conducted in Korea. To evaluate the presence of ARV infection in broiler farms, epidemiological survey such as serological test and virus isolation has been conducted. For the serological survey using ELISA method, we selected five broiler farms which were located at different area and had a history of growth retardation, lameness, diarrhea and poor feathering. From these farms serum samples were collected at 1 day, 14 days and market age. All these farms had no history of vaccination against ARV. In addition to serological survey, we tried to isolate ARV from birds of designated farms at market age and collected feces and tissue samples such as cecal tonsil, intestine and liver. We were identified ARV by RT-PCR and transmissible electron microscopy. The samples were inoculated into 9-day-old embryonated eggs via the chorioallantoic membrane to observe the pock formation. For the pathogenicity test of ARV isolates, we inoculated with the isolates to the right footpad of 3-week-old SPF chicks and observed clinical signs and pathological changes for 14 days after challenge. Most broilers sampled for serological survey have maternal antibodies which were widely distributed at 1 day and decreased by 14 days. However, at the market age several broiler farms showed fairly high antibody titer against ARV. This increase of antibody titer at market age means the possible infection of ARV during the grow-out period. Among total 15 samples for the isolation of ARV. 2 samples were positive by RT-PCR and finally identified as a ARV. We inoculated these isolates in the SPF birds and observed that the antibody titer was increased from 7 days after challenge. However, we did not find any clinical signs both control and challenge groups. Based on the above results, it is clear that the ARV infection has been circulated in the broiler industry and caused significant economic losses. Further study is needed to evaluate the virulence of the isolates in the digestive system of broiler and the molecular characteristics of isolates.

• Korean Journal of Ecology and Environment
• /
• v.41 no.3
• /
• pp.402-411
• /
• 2008

Grazing effects of two freshwater shellfish, Unio douglasiae (UNIO) and Cipangoplaudina chinese malleata (CCM), on the hibernal diatom communities in eutrophic water was examined in a laboratory. Two animals having different feeding types used in the study were collected from Keum River (Boryeong, Korea), acclimatized in the artificial management system in a laboratory over at least one month, and starved in a laboratory for 2 days before the experiment. Experimental waters, which dominated by Synedra ulna-Scenedesmus sp. (ca. 80%), was collected from eutrophic lake, Lake Ilgam (Seoul) in Feb., 19, 2008 at AM 10:00, and used in the study after the filtration with 1 mm Nylon mesh. Feeding experiments were largely divided into three kinds of animal treatments; five densities of UNIO (U0, U1, U2, U3, U4) and CCM (C0, C1, C2, C3, C4), and four combined densities of two shellfish (M0, M1, M2, M3). U0 and C0 were control (no addition of UNIO or CCM), U1 or C1 (each animal density at 0.5 ind. $L^{-1}$), U2 or C2 (1.0 ind. $L^{-1}$), U3 or C3 (1.5 ind. $L^{-1}$), and U4 or C4 (2.0 ind. $L^{-1}$), respectively. Four combined treatments were consisted of M0 (no animals), M1 (U1+C3), M2 (U2+C2), and M3 (U4+C1), respectively. Under the presence of animal, the concentration of Chl-a and algal abundance were clearly decreased with the increase of UNIO density and the treated time, while in combined group a strong decrease of algal density showed with the increase of UNIO density. Total phytoplankton density shifted as the similar patterns to that of Chl-a concentration (r=0.6527, p<0.0001), however, there showed the differences following a species. There were strong decreases of dominant species Synedra ulna, Scenedesmus sp., Ankistrodesmus falcatus in UNIO treatment group, Diatoma vulgare in combined group, while Cryptomonas ovata in high density of CCM increased about 20% in algal density. Grazing rates (GRs) based on the concentration of Chl-a was depend on the kind of shellfish and treatment time; a strong feeding of CCM showed in the initial stage, and four hours later, UNIO and combined group with high UNIO density showed the high GRs. Interestingly, faeces production of shellfish was highest in combined group with high density of CCM, while their size over 60 ${\mu}m$ was much higher in production magnitude than that less 60 ${\mu}m$. Collectively, these results suggest that two domestic shellfish and its combined treatment have a strong potential as an effective biological controller of diatom bloom in cold eutrophic waters.

• The Korean Journal of Nuclear Medicine Technology
• /
• v.16 no.2
• /
• pp.7-11
• /
• 2012

Purpose : The effect of concomitant use of $^{18}F$-FDG and intravenous contrast agent (CA) on dual-energy X-ray absorptiometry (DXA), was rarely reported. We had investigated these potentially confounding effects. Materials and Methods : Twenty-two patients had undergone DXA before and immediately after $^{18}F$-FDG PET/CT scans. Two DXA and 1 PET/CT scans had performed within one-day. $^{18}F$-FDG PET/CT scans had been performed with CA in 17 patients and without CA in 5 patients. Whole body bone mineral content (BMC), whole body bone mineral density (BMD), total fat mass (TFM), and lean body mass (LBM) were measured by DXA scanner before and after the $^{18}F$-FDG PET/CT scans. Results : BMC, BMD, TFM and LBM had significantly affected by $^{18}F$-FDG PET/CT with CA (BMC, +13.7%, from $2061.3{\pm}393.7$ to $2343.4{\pm}373.3$; BMD, +9.3%, from $1.07{\pm}0.09$ to $1.17{\pm}0.08$; TFM, -34.1%, from $17052.1{\pm}4049.9$ to $11237.1{\pm}2990.3$; LBM, +13.6%, from $45834.5{\pm}5662.1$ to $52094.0{\pm}6335.4$). However, $^{18}F$-FDG PET/CT without CA had no effect on the measurement of DXA (BMC, +2.4%, from $2197.7{\pm}391.6$ to $2251.5{\pm}380.9$; BMD, +1.8%, from $1.13{\pm}0.09$ to $1.15{\pm}0.07$; TFM, -6.8%, from $14585.6{\pm}3455.9$ to $13591.3{\pm}4351.4$; LBM, +2.2%, from $47360.5{\pm}8381.8$ to $48441.1{\pm}8488.1$). Conclusion : The measurements of DXA are affected by using CA. However, DXA scans might be unaffected by the presence of $^{18}F$-FDG administered for PET/CT.

• The Korean Journal of Nuclear Medicine
• /
• v.34 no.5
• /
• pp.403-409
• /
• 2000

Objectives: Due to the heterogeneous receptor distribution and changes of receptor status over time, the biochemical measurement of estrogen receptor status of biopsy specimens is not sufficient to diagnose breast cancer. As a result, I-123 labeled estradiols have been applied for the diagnosis. The purpose of this study was to develop a suitable radioligand for imaging estrogen receptor-positive human breast tumors. Methods: Among the various estradiol derivatives, $17{\alpha}-[^{123}I]$iodovinyl estradiol ($[^{123}I]$IVE) has been prepared from $17{\alpha}$-ethynyl estradiol. Labeling of $E-17{\alpha}-[^{123}I]$iodovinyl estradiol (E-$[^{123}I]$IVE) was carried out using peracetic acid with $[^{123}I]NaI\;and\;Z-[^{123}I]IVE$ labelling was archived using chloamine-T/HCl solution with $[^{123}I]$NaI. Labeling yield was determined by silica thin-layer chromatography (TLC) and radiochemical purity was measured by high performance liquid chromatography (HPLC). The biodistribution of E-$[^{123}I]$IVE was measured in immature female rats at 60 min, 120 min and 300 min after injection. Results: The labeling yield of two isomers was 92% and 94% ($E-[^{123}I]IVE\;and\;Z-[^{123}I]IVE$, respectively). The radiochemical purity was more than 98% after purification. The highest uptake was observed at 120 min in uterus (3.11% ID/g for E-$[^{123}I]$IVE). Conclusion: These results suggest the possibility of using E-$[^{123}I]$IVE as an imaging agent for the evaluation of the evaluation of the presence of estrogen receptor in patients with breast cancer.

• Korean journal of applied entomology
• /
• v.31 no.4
• /
• pp.496-508
• /
• 1992

Phytophagous insects associated with Dicotyledoneae weeds and host specificities in the field populations were investigated for the survey of biological control agents of weeds in Korea. Fifty four weed species in 39 genera were collected during the survey. The most insects were collected from Polygonales by 24 species in 22 genera and followed by Urticales and Centrospermales by 17 species of 17 genera. The insects collected in the other weed orders were ranged from 1 to 12 species. Out of 17 insect species collected in Urticales, Baris sp. damaged the leaves of Hamulus japonicus in Cannabinaceae as scattered holeshape and showed host specificity. In Polygonaceae, Rumex japonicus and R. crispus were severely damaged by Aphis rumicis and Gastrophysa atrocyanea. G. atrocyanea leaf beetle had host specificity on R. japonicus and ate all the leaves except veins. The leaf beetle, Lypesthes japonicus was a potential biological control agent by feeding leaves of Persicaria spp .. And Lixus spp. were also often collected from Persicaria spp .. Liothrips vaneeckei was first collected from weed, P. modosa. P. senticosa was damaged by unidentified geometrid moth larvae and P. perfoiliata by Miarus atricolor snout beetle. Cassida piperata damaged leaves of Chenopodium album of Centrospermales and showed host specificity. In a soybean field, C. album and Amaranthus mangostanus were severely damaged by Spodoptera litura larvae which were eating soybean leaves. This phenomenon indicates that the presence of weed in cultivated land influences the outbreak of insect pests. Altica oleracea leaf beetle was frequently collected from Oenothera spp. of Onagraceae in Myrtales. Aphis gossyphi was outbroken on Solanum nigrum and Phylliodes brettinghami leaf beetle was first recorded on the same plant. Leaf beetles, Longitarsus scutellais and Hemipyxis plagioderoides were first collected from Plantago asiatica of Plantaginaceae in Plantaginales. They showed host specificities in the fields. The hemipterans were collected from many weeds during the survey and their roles on weeds should be investigated. A tractomorpha bedeli was also collected from many kinds of weeds in forest areas.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.29 no.1
• /
• pp.147-152
• /
• 2000

Various lines of evidence suggest that dietary components protect the initiation of carcinogenesis. In this study, the ethanol extracts (AGE) and the methanol and hexane partition layers (AGEM, AGEH) of the Angelica radix were screened for their cytotoxic effects using the MTT assay on HepG2, HeLa, MCF7 and SW626 cells and for their ability to induce quinone reductase (QR) in HepG2 cells. AGEM and AGEH of the Angelica radix showed the strongest cytotoxic effects on HepG2 and HeLa cells. Cell growth was inhibited by 99.8% and 99.8% on HepG2 cells and 99.3% and 99.4% on HeLa cells, at dose of $100\;\mu\textrm{g}/ml$ of AGEM and AGEH extracts respectively. AGE and AGEH significantly induced QR activities in the HepG2 cells. The QR activities of HepG2 cells grown in the presence of AGE, AGEH, and AGEM at the concentration of $50\;\mu\textrm{g}/mL$ were 313.5, 273.3 and 133.3 nmol/min/mg protein, respectively. Therefore, based on these studies, Angelica radix may be developed into a potentially useful cancer chemopreventive agent.

• Archives of Pharmacal Research
• /
• v.21 no.4
• /
• pp.458-464
• /
• 1998

Search for a new $\alpha$-methylene-$\gamma$-butyrolactone-bearing 6-substituted purine as a potental antitumor agent has led to synthesize seven, hitherto unreported, $5^1$-Methyl-$5^1$-[(6-substituted-9H-purin-9-yl)methyl]-$2^1$-oxo-$3^1$- methylenetetrahydrofurans (H, Cl, l, $CH_3$, $NH_2$, SH, >C=O) (6a-g). These include $5^1$-Methyl-$5^1$-[(9H-purin-9-yl)methyll-$2^1$-oxo-$3^1$ -methylenetetrahydrofurans (6a), $5^1$-Methyl-$5^1$-[(6-chloro-9H-purin-9-yl)methyl]-$2^1$-oxo-$3^1$-methylenetetrahydr ofurans (6b), $5^1$-Methyl-$5^1$-[(6-chloro-9H-purin-9-yl) methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofurans (6c), $5^1$-Methyl-$5^1$-[(6-methyl-9H-purin-9-yl) methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofurans (6d), $5^1$-Methyl-$5^1$-[(9H-adenin-9-yl)methyll-$2^1$-oxo-$3^1$-methylenetetrahydrofurans (6e), $5^1$-Methyl-$5^1$-[(6-mercapto-9H-purin-9-yl) methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofurans (6f) and $5^1$-Methyl-$5^1$-[(9H-hypoxanthin-9-yl)methyll-$2^1$-oxo-$3^1$-methylenetetrahydrof urans (6g) which were made by the Reformatsky-type reaction of ethyl $\alpha$-(bromomethyl) acrylate with the corresponding (6-substituted-9H-purin-9-yl)-2-propanone intermediates (5a-g). These ketone intermediates 5a-g, 1-(9H-purin-9-yl)-2-propanone (5a), 1-(6-chloro-9H-purin-9-yl)-2-propanone (5b), 1-(6-iodo-9H-purin-9-yi)-2-propanone (5c), 1-(6-methyl-9H-purin-9-yl)-2-propanone (5d), 1-(9H-adenin-9-yl)-2-propanone (Se), 1-(6-mercapto-9H-purin-9-yl)-2-propanone (5f), and 1-(9H-hypoxanthin-9-yl)-2-propanone (5g) were directly obtained by the alkylation of the 6-substituted purine bases with the chloroacetone in the presence of $K_2$$CO_3$ (or NaH) under DMF (or DMSO). The preliminary in vitro cytotoxcity assay for the synthetic .alpha.-methylene-y-butyro-lactone compounds (6a-g) were determined against three cell lines (PM-3A, P-388, and K-562) and showed the moderate antitumor activity ($IC_50$ ranged from 1.4 to 4.3 $\mu\textrm{g}$/ml) with the compound $5^1$-methyl-$5^1$ -[(9H-hypoxanthin-9-yl)methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofuran (6g) showing the least antitumor activity.

• Applied Biological Chemistry
• /
• v.22 no.2
• /
• pp.109-122
• /
• 1979

Present study was carried out to reduce residual toxicity of BHC insecticides inherent in the organochlorine pesticides. For This end, r-isomer, the most potent insecticidal component among the BHC stereoisomers, was isolated and thus fortified by means of solvent precipitation. In parallel, 3-(2,4,5-trichlorophenyl)-1-methyl urea was prepared in good yield from technical BHC via 1,2,4-trichlorobenzene, 1,2,4,-trichloronitrobenzene, and 2,4,5-trichloroaniline. In addition, certain merit of the compound which make it possible to use as a herbicide is discussed. The results are summarized as follows; 1. Recrystallizing technical BHC from methanol-water binary solvent system, r-isomer was enriched to 49.7% at 95% recovery of r-isomer. 2. By partitioning technical BHC in 85% of methanolic solution into chloroform, r-isomer was fortified to 89.6% at 90.5% recovery of r-isomer. 3. Yield of 1,2,4-trichlorobenzene from technical BHC was greatly dependent upon concentration of alkalies and to less degree on the alkalies. 4. Surfactants, in particular cationic a quartenary ammonium salt, increased yield of 1,2,4-trichlorobenzene from technical BHC by alkaline hydrolysis. 5. Conversion of 1,2,4-trichlorobenzene to 2,4,5-trichloronitrobenzene was effected almost quantitatively utilizing $HNO_3-H_2SO_4$ nitrating agent at low temperature. 6. Yield of 91.4% was observed for the synthesis of 2,4,5-trichloroaniline by reducing 2,4,5-trichloronitrobenzene in the presence of iron turning and hydrochloric acid. 7. Overall yield based on BHC of 3-(2,4,5-trichlorophenyl)-1- methyl urea was 60.8%. 8. Inhibition effects, both germination and growth, 3-(2,4,5-trichlorophenyl)-1-methyl urea on several crops were found comparable to or more potent than those of $linuron{\circledR}\;and\;diuron{\circledR}$. In addition, it was also noted that susceptibility to the prepared compound depended upon the crops as well as specific part (shoots, roots) of the plant exposed to the chemicals.

• Journal of Life Science
• /
• v.23 no.2
• /
• pp.197-206
• /
• 2013

The present study investigated whether leukemia-maintaining cells reside in a differentiation-resistant fraction using a megakaryocytic differentiation model of K562 cells. Treatment with phorbol-12-myristate-13-acetate (PMA) significantly inhibited the colony-forming efficiency of the K562 cells. At a PMA concentration of 1 nM or higher, colony was not formed, but approximately 40% of K562 cells still survived in soft agar. Approximately 70% of colony-forming cells that were isolated following the removal of PMA after exposure to the agent were differentiated after treatment with 10 nM PMA for 3 days. The differentiation rate of the colony-forming cells was gradually increased and reached about 90% 6 weeks after colony isolation, which was comparable to the level of a PMA-treated K562 control. Meanwhile, imatinib-resistant variants from the K562 cells, including K562/R1, K562/R2, and K562/R3 cells, did not show any colony-forming activity, and most imatinib-resistant variants were CD44 positive. After 4 months of culture in drug-free medium, the surface level of CD44 was decreased in comparison with primary imatinib-resistant variants, and a few colonies were formed from K562/R3 cells. In these cells, Bcr-Abl, which was lost in the imatinib-resistant variants, was re-expressed, and the original phenotypes of the K562 cells were partially recovered. These results suggest that leukemia-maintaining cells might reside in a differentiation-resistant population. Differentiation therapy to eliminate leukemia-maintaining cells could be a successful treatment for leukemia if the leukemia-maintaining cells were exposed to a differentiation inducer for a long time and at a high dose.