Purpose: The herpes simplex virus type 1 thymidine kinase gene(HSV1-tk) is an attractive candidate as a reporter gene in noninvasive reporter gene monitoring system. The HSV1-tk gene was chosen as a reporter gene, because it has been extensively studied, and there are appropriate reporter probes, substrates of HSV1-tk gene product, to apply for HSV1-tk gene imaging. We used radiolabeled 5-iodovinyl-2'-deoxyuridine (IVDU) and 5-iodovinyl-2'-fluoro-2'-deoxyuridine (IVFRU) as reporter probes for HSV1-tk gene monitoring system. Materials and Methods: We prepared HSV1-tk gene transduced Morris hepatoma cell line using retroviral vector, MOLTEN containing HSV1-tk gene. And we confirmed the HSV1-tk gene expression by Northern blotting and Western blotting. We compared in vitro uptakes of radioiodinated IVDU and IVFRU to monitor HSV1-tk gene expression in Morris hepatoma cell line (MCA) and HSV1-tk gene tranduced MCA (MCA-tk) cells until 480 minutes. We also peformed correlation analysis between percentage of HSV1-tk gene tranduced MCA cell % (MCA-tk%) and uptakes of radiolabeled IVDU or IVFRU. Results: MCA-tk cell expressed HSV1-tk mRNA and HSV1-TK protein. Two compounds showed minimal uptake in MCA, but increased uptake was observed in MCA-tk. IVDU showed 4-fold higher accumulation than IVFRU at 480 min in MCA-tk (p<0.01). Both IVDU and IVFRU uptake were linearly correlated ($R^2>0.96$) with increasing MCA-tk%. Conclusion: The radiolabeld IVDU and IVFRU showed higher specific accumulation in retrovirally HSV1-tk gene transfected Morris hepatoma cell line. Both IVDU and IVFRU could be used as good substrates for evaluation of HSV1-tk gene expression.
Avermectin (AVM) $B_{1a}$ produced by Streptomyces avermitilis via polyketide pathway is a secondary metabolite with powerful anthelmintic and insecticidal activities, thus being used as an efficient agent in the field of agriculture and animal health. It has been reported that a precursor for AVM $B_{1a}$ biosynthesis was isoleucine and the biosynthetic pathway of AVM $B_{1a}$ was closely similar to that of fatty acid. Based on understanding of the biosynthetic pathway of AVM $B_{1a}$, we intended to screen various mutants resistant against O-methyl threonine (OMT), an isoleucine-anti metabolite, and/or mutants resistant against p-fluoro phenoxy acetic acid (pFAC), an inhibitor of fatty acid biosynthesis. It was inferred that these mutants could produce AVM $B_{1a}$ more efficiently, due to the acquired capability of not only overproducing isoleucine intracellularly but also channelling metabolized carbon-sources into the polyketide pathway, thus leading to enhanced biosynthesis of AVM $B_{1a}$. The resulting mutant (PFA-1 strain) resistant against 100 ppm of pFAC was able to produce approximately 42 fold higher amount of AVM $B_{1a}$ compared to the parallel mother strain (4,200 vs. 100 units/l). In addition, through the process of continuous strain improvement program carried out by gradually increasing the OMT concentration, it was possible to obtain a more attractive mutant with greater AVM $B_{1a}$ production capacity (9,000 units/l). Notable was that significantly higher producer (12,000 units/l) could be selected through further screening of the resistant mutants, this time, to even higher concentration of PFAC. Meanwhile, through the analysis of AVM Bla production histograms (i.e., number of strains according to their AVM $B_{1a}$ biosynthetic ability) for the earlier strains in comparison with the high producers having the characteristics of resistance to OMT and pFAC, it was found that production stability of the high-yielding producers were remarkably improved, as demonstrated by the fact that larger proportion of the mutated strains had greater capability of AVM $B_{1a}$ biosynthesis ($71\%$ in the range between 5,000 and 7,000 units/L; $47\%$ in the range between 6,000 and 7,000 units/l). Based on these consequences, it was concluded that the rational screening strategy based on the understanding of the biosynthetic pathway of AVM $B_{1a}$ was very effective in obtaining high-yielding mutants with the features of enhanced production stability.
Lee Sang-wook;Im Ki Chun;Nam Soon Yuhl;Kim Jae Seung;Choi Eun Kyung;Ahn Seung Do;Shin Seong Soo;Ryu Jin Sook;Kim Sang Yoon;Lee Bong-Jae;Choi Seung-Ho;Kim Sung-Bae;Moon Dae Hyuk
Radiation Oncology Journal
/
v.23
no.1
/
pp.9-16
/
2005
Purpose : To prospectively evaluate the use of positron emission tomography with the glucose analog fluoro-deoxyglucose (FDG-PET) to deoxyglucose (FDG-PET) to predict disease-free survival (DFS) after concurrent chemo-radiotherapy (CCRT) in patients with non-disseminated nasopharyngeal carcinoma (NPC). Materials and Methods : We studied 41 patients with non-disseminated NPC scheduled to undergo platinum-based CCRT were eligible for this study. Patients were studied by FDG-PET prior to the CCRT. FDG uptake of tumors were measured with the maximal standardized uptake value (SUV). Results : Complete response rate was $100\%$. In ten patients who presented with any component of treatment failure, the median $SUV_{max}$ was 8.55 (range: $2.49\~14.81$) in any component of failure and the median $SUV_{max}$ was 5.48 (range: $2.31\~26.07$) In the remaining patients without any such failure. Patients having tumors with high FDG uptake had a significantly lower 3-year DFS ($51\%\;{\nu}91\%$, p=0.0070) compared with patients having low uptake tumors. Conclusion : FDG uptake, as measured by the SUV, has potential value in predicting DFS in NPC treated by CCRT, High FDG uptake may be a useful parameter for Identifying patients requiring more aggressive treatment approaches.
Purpose: The aim of this study was to investigate the feasibility of 3 ' -[F-18]fluoro-3 ' -deoxythymidine positron emission tomography(FLT-PET) for the detection of locally advanced breast cancer and to compare the degree of FLT and 2' -deoxy-2 ' -[F-18]fluoro-d-glucose(FDG) uptake in primary tumor, lymph nodes and other normal organs. Material & Methods: The study subjects consisted of 22 female patients (mean age; $42{\pm}6$ years) with biopsy-confirmed infiltrating ductal carcinoma between Aug 2005 and Nov 2006. We performed conventional imaging workup, FDG-PET and FLT PET/CT. Average tumor size measured by MRI was $7.2{\pm}3.4$ cm. With visual analysis, Tumor and Lymph node uptakes of FLT and FDG were determined by calculation of standardized uptake value (SUV) and tumor to background (TB) ratio. We compared FLT tumor uptake with FDG tumor uptake. We also investigated the correlation between FLT tumor uptake and FDG tumor uptake and the concordant rate with lymph node uptakes of FLT and FDG. FLT and FDG uptakes of bone marrow and liver were measured to compare the biodistribution of each other. Results: All tumor lesions were visually detected in both FLT-PET and FDG-PET. There was no significant correlation between maximal tumor size by MRI and SUVmax of FLT-PET or FDG-PET (p>0.05). SUVmax and $$SUV_{75} (average SUV within volume of interest using 75% isocontour) of FLT-PET were significantly lower than those of FDG-PET in primary tumor (SUVmax; $6.3{\pm}5.2\;vs\;8.3{\pm}4.9$, p=0.02 /$SUV_{75};\;5.3{\pm}4.3\;vs\;6.9{\pm}4.2$, p=0.02). There is significant moderate correlation between uptake of FLT and FDG in primary tumor (SUVmax; rho=0.450, p=0.04 / SUV75; rho=0.472, p=0.03). But, TB ratio of FLT-PET was higher than that of FDG-PET($11.7{\pm}7.7\;vs\;6.3{\pm}3.8$, p=0.001). The concordant rate between FLT and FDG uptake of lymph node was reasonably good (33/34). The FLT SUVs of liver and bone marrow were $4.2{\pm}1.2\;and\;8.3{\pm}4.9$. The FDG SUVs of liver and bone marrow were $1.8{\pm}0.4\;and\;1.6{\pm}0.4$. Conclusion: The uptakes of FLT were lower than those of FDG, but all patients of this study revealed good FLT uptakes of tumor and lymph node. Because FLT-PET revealed high TB ratio and concordant rate with lymph node uptakes of FDG-PET, FLT-PET could be a useful diagnostic tool in locally advanced breast cancer. But, physiological uptake and individual variation of FLT in bone marrow and liver will limit the diagnosis of bone and liver metastases.
The purpose of this study was to identify the preventive and the progressive inhibitory effects of enamel demineralization with fluoride releasing light-and self-cured orthodontic sealants(FluoroBond), in vitro, under the polarizing light microscope and the scanning electon microscope. The polarizing light microscopic group was subdivided into seven groups(Group A-Group G). The scanning electron microscopic group was also subdivided into seven groups(Group A'-Goup G'). For polarizing light microscopic evaluation, longitudinal sections were made longitudinally by Maruto cutter(Maruto Co., Japan) and Maruto grinding machine(Maruto Co., Japan). Sections were examined and photographed by the polarizing light microscope(Olympus Optical Co., Japan) using crossed polars and with the enamel rod longitudinal axis oriented at $45^{\circ}$ to the extinction position. For scanning electron microscopic evaluation, the specimens were coated with a highly conducting layer of gold palladium in a model Hus-4 high-vacuum evaporator and examined in an ISI-100B scanning electron microcope operated at 20kV. The results of this study were as follows : 1. The mean depths of artificial carious lesions under a polarized light microscope were $Group\;A(5.08{\mu}m),\;Group\;B(47.82{\mu}m,\;Group\;C(8.42{\mu}m),\;Group\;D(7.20{\mu}m),\;Group\;E(85.41{\mu}m),\;Group\;F(60.38{\mu}m),\;Group\;G(60.13{\mu}m)$. 2. There were statistically significant differences in Group B compared with Group A, C, and D(p<0.05), and also, in Group I compared with Group F and Group G(p<0.05). 3. Light-and self-cured orthodontic sealants had the preventive effects of enamel demineralization. 4. Light-and self-cured orthodontic sealants had the progressive inhibitory effects of enamel demineralization. 5. The time progress of demineralizing agent had no influence on the samples of light-and self-cured orthodontic sealants under the scanning electron microscope. 6. There was no difference between the specimens of light-and self-cured orthodontic sealants both in the polarized light microscopic group and in the scanning electron microscopic group.
Background: High 2-[$^{18}F$] fluoro-2-deoxy-D-glucose (FDG) uptake on positron emission tomography-computed tomography (PET-CT) is a prognostic factor for poor survival in non-small cell lung cancer (NSCLC), especially in Stage I. We determined whether the high FDG uptake value of a primary tumor was associated with recurrence and death in patients with resected Stage I and Stage II NSCLC. Methods: We identified consecutive patients who underwent complete surgical resection for Stage I and II NSCLC between 2006 and 2009, who had preoperative PET-CT, and reviewed clinical records retrospectively. FDG uptake was measured as the maximal standardized uptake value (SUVmax) for body weight. Patients were divided into two groups based on SUVmax: (i) above or (ii) below the cut-off value (SUVmax=5.9) determined by a receiver operating characteristic (ROC) curve. Results: Of 57 patients who were enrolled consecutively, 32 (56%) had Stage I NSCLC and 25 (44%) had Stage II. The 5-year recurrence-free survival (RFS) for patients with high (${\geq}5.9$) and low (<5.9) SUVmax were 31% and 57%, respectively (p=0.014). The 5-year overall survival (OS) rates were 39% and 60%, respectively (p=0.029). In univariate analyses, SUVmax (p=0.014), T staging (p=0.025), and differentiation of tumor tissue (p=0.034) were significantly associated with RFS. But, multivariate analyses did not show that SUVmax was an independently significant factor for RFS (p=0.180). Conclusion: High FDG uptake on PET-CT is not an independent prognostic factor for poor outcomes (disease recurrence in patients with resected Stage I and II NSCLC).
In order to develope a clean technology by liquid-supported membrane of ionic liquid/PVDF, the solubility of R22 and HFP gases using BMIBF4 as ionic liquid were measured at temperatures from 0 to $30^{\circ}C$, at total pressures up to 4 bars. The solubility of R22 in this ionic liquid was shown a rapid increasing tendency with increases of pressure and decreases of temperature, respectively, whereas the solubility of HFP was showed only a little in the same conditions. Based on these results, liquid-supported membranes of ionic liquid/PVDF were prepared by variables of the deposition amount of ionic liquid in polymer matrix, PVDF and were applied to the separation of fluoro-gases(R22, HFP) including $N_2$ gas. The permeability of R22 was rapidly increased by depending on the deposition amounts of ionic liquid, whereas both of HFP and $N_2$ were just showed so little. Especially, the diffusivity coefficient and solubility parameter of R22 were increased by lower operating temperatures and increased deposition amount of ionic liquid in 1iquid membrane. In conclusion, the selectivity of R22 against HFP was changed to 10-45 times depending on both of operating temperatures and the deposition amount of ionic liquid in BMIBF4/PVDF liquid membrane.
Sohn, Young Bae;An, Young Sil;Lee, Su Jin;Choi, Jin Wook;Jeong, Seon-Yong;Kim, Hyon-Ju;Ko, Jung Min
Journal of Genetic Medicine
/
v.9
no.2
/
pp.84-88
/
2012
Purpose: Neurofibromatosis type 1 (NF1), which is caused by mutations of the NF1 gene, is the most frequent single gene disorder to affect the nervous system. Unidentified bright objects (UBOs) are commonly observed on brain magnetic resonance imaging (MRI) in patients with NF1. However, their clinical and pathologic significance is not well understood. The purpose of this study was to investigate the correlation between UBOs and cerebral glucose metabolism measured by $^{18}F$-2-Fluoro-2-deoxy-D-glucose ($^{18}F$-FDG) positron emission tomography (PET) in Korean patients with NF1. Materials and Methods: Medical records of 75 patients (34 males and 41 females) with NF1 who underwent brain MRI and PET between 2005 and 2011 were evaluated retrospectively. Clinical data including demographics, neurological symptoms, and brain MRI and PET findings, were reviewed. Results: UBOs were detected in the brain MRI scans of 31 patients (41%). The region most frequently affected by UBOs was the basal ganglia. The most frequent brain PET finding was thalamic glucose hypometabolism (45/75, 60%). Of the 31 patients with UBOs, 26 had thalamic glucose hypometabolism on brain PET, but the other 5 had normal brain PET findings. Conversely, of the 45 patients with thalamic glucose hypometabolism on brain PET, 26 showed UBOs on their brain MRI scans, but 19 had normal findings on brain MRI scans. Conclusion: UBOs on brain MRI scans and thalamic glucose hypometabolism on PET appear to be 2 distinctive features of NF1 rather than correlated symptoms. Because the clinical significance of these abnormal imaging findings remains unclear, a longitudinal follow-up study of changes in clinical manifestations and imaging findings is necessary.
The pharmacokinetics and tissue distribution of DWP20367 (1-cyclopropyl-6-fluoro-8-chloro-7-(2, 7-diazabicyclo[3,3,0]tract-4-ene-7-yl)-1,4-dihydro-4-oxoquinoline-3-carboxylic acid), a novel fluoroquinolone, were examined in rats and beagle dogs after a single intravenous and oral administration. Analysis of DWP20367 in plasma, tissue, and urine was determined by both HPLC and microbiological assay (bioassay). The plasma concentration-time curves of the drug in rats and beagle dogs were biexponentially declined. The terminal half-life (t$_{1}$2$\beta$/) of the drug in rats was about 60.1 $\pm$7.3 min (i.v.) and 61.3 $\pm$ 12.4 min (p.o.) in bioassay, and 86.3 $\pm$19.8 min (i.v.) and 50.9$\pm$ 14.9 min (p.o.) in HPLC. In beagle dogs, half-life of the drug determined by bioassay was about 121.8$\pm$6.2 min (i.v.) and 111.0$\pm$7.6 min (p.o.). The volume of distribution at steady-state (Vd$_{ss}$ ) was 243.8$\pm$74.1 ml/kg (bioassay) and 339.2$\pm$84.3 ml/kg (HPLC) in rats, and 1587.5 $\pm$536.9 ml/kg (bioassay) in beagle dogs. The total body clearance (Cl$_{t}$) of DWP20367 was 3.4 $\pm$ 0.4 ml/min/kg (bioassay) and 2.4$\pm$0.4 ml/min/kg (HPLC) in rats, and 12.3$\pm$ 1.0 ml/min/kg (bioassay) in beagle dogs, respectively. The extent of bioavailability after oral administration was 89.1%(bioassay) and 79.9% (HPLC) in rats, and 78.7% (bioassay) in beagle dogs. Urinary recovery (24-h) assayed by bioassay was 0.7% (p.o.) and 1.2% (i.v.) in rats, and 0.8% (p.o.) and 1.0% (i.v.) in beagle dogs. In rats, 24-h fecal recovery determined by bioassay was 11.2% (p.o.) and 0.1% (i.v.). Rat and human serum protein binding ratios at 2$\mu$g/ml were about 90~91%. This drug determined by bioassay was also distributed by the order of liver, kidney, lung, heart, spleen and muscle 30 min after oral administration.on.
Park, Jun-Hyung;Im, Ki-Seop;Lee, Hong-Jin;Jeong, Kyung-Il;Lee, Byung-Chul;Lee, In-Won
The Korean Journal of Nuclear Medicine Technology
/
v.13
no.3
/
pp.147-151
/
2009
Purpose: 2-[$^{18}F$]Fluoro-2-deoxy-D-glucose ([$^{18}F$]FDG) particularly plays as a important role in Positron Emission Tomography (PET) imaging in nuclear medicine. Domestic [$^{18}F$]FDG auto synthesizers are installed in Seoul National University Bundang Hospital (SNUBH) at June 2008, these modules were known that it's synthetic yields were guaranteed in average $45{\pm}5%$ so far. To improve yields and convenience of domestic [$^{18}F$]FDG auto synthesizer, numerous trials in reaction time, base concentration, pressure and temperature were performed to increase [$^{18}F$]FDG yields. Materials and Methods: Several synthetic factors (temperature, time and pressure) and shortcoming were corrected based on many evaporation test. Syringe dispensing of tetra-butylammonium bicarbonate (TBAB) was replaced with micro pipette to prepare tetrabutyl ammonium fluoride salt ([$^{18}F$]TBAF). Troublesome refill of liquid nitrogen every 2 hours which was used to protect vacuum system was changed to charcoal cartridge, base guard filter. To monitor the volume of delivered $[^{18}O]OH_2$ from cyclotron by surveillance camera, we set up the volumetric vial on the cover of the module. In addition to, the recovery vial was added in [$^{18}F$]FDG production system to recover [$^{18}F$]FDG loss due to the leak of valve ($V_{13,14}$) in [$^{18}F$]FDG purification process. Results: When we used micro pipette for adding TBAB ($30\;{\mu}L$ in 12% $H_2O$ in acetonitrile), this quantitative dispensation has enabled to improve $5.5{\pm}1.7%$ residual fluorine-18 activity in fluorine separation cartridge compared to syringe adding. Besides, the synthetic yields of [$^{18}F$]FDG has increased $58{\pm}2.6%$ (n=19), $58{\pm}2.9%$ (n=14), $60%{\pm}2.5%$ (n=17) for 3 months. The life cycle of charcoal cartridge and base vacuum was 3 months prior to filling liquid nitrogen every 2 hours and additional side separator can prevent pump corrosion by organic solvent. After setting of volumetric indicator vial, the operator can easily monitor the total volume of irradiated $[^{18}O]OH_2$ from cyclotron. The recovery vial can be used for the stabilizer when an irregular [$^{18}F$]FDG loss was generated by the leak of valves ($V_{13,14}$). Conclusions: We has optimized appropriate synthetic conditions (temperature, time, pressure) in domestic [$^{18}F$]FDG auto synthesizer. In addition to, the remodeling with several accessories improve yields of domestic [$^{18}F$]FDG auto synthesizer with reliable reproducibility.
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