• Nuclear Medicine and Molecular Imaging
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• v.42 no.3
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• pp.235-245
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• 2008

Purpose: The HSV1-tk gene has been extensively studied as a type of reporter gene. In hepatocellular carcinoma (HCC), only a small proportion of patients are eligible for surgical resection and there is limitation in palliative options. Therefore, there is a need for the development of new treatment modalities and gene therapy is a leading candidate. In the present study, we investigated the usefulness of substrate, 2'-fluoro-2'-deoxy-1-${\beta}$-D-arabino-furanosyi-5-[$^{124/125}I$]iodo- uracil ([$I^{124/125}I$]FIAU) as a non-invasive imaging agent for HSV1-tk gene therapy in hepatoma model using small animal PET. Material and Methods: With the Morris hepatoma MCA cell line and MCA-tk cell line which was transduced with the HSV1-tk gene, in vitro uptake and correlation study between [$^{125}I$]FIAU uptake according to increasing numeric count of percentage of MCA-tk cell were performed. The biodistribution data and small animal PET images with [$^{124}I$]FIAU were obtained with Balb/c-nude mice bearing both MCA and MCA-tk tumors. Results:, Specific accumulation of [[$^{125}I$]FIAU was observed in MCA-tk cells but uptake was low in MCA cells. Uptake in MCA-tk cells was 15 times higher than that of MCA cells at 480 min. [$^{125}I$]FIAU uptake was linearly correlated (R2 =0.964, p =0.01) with increasing percentage of MCA-tk numeric cell count. Biodistribution results showed that [$^{125}I$]FIAU was mainly excreted via the renal system in the early phase. Ratios of MCA-tk tumor to blood acting were 10, 41, and 641 at 1 h, 4 h, and 24 h post-injection, respectively. The maximum ratio of MCA-tk to MCA tumor was 192.7 at 24 h. Ratios of MCA-tk tumor to liver were 13.8, 66.8, and 588.3 at 1 h, 4 h, and 24 h, respectively. On small animal PET, [$^{124}I$]FIAU accumulated in substantial higher levels in MCA-tk tumor and liver than MCA tumor. Conclusion: FIAU shows selective accumulation to HSV1-tk expressing hepatoma cell tumors with minimal uptake in normal liver. Therefore, radiolabelled FIAU is expected to be a useful substrate for non-invasive imaging of HSV1-tk gene therapy and therapeutic response monitoring of HCC.

• Korean Journal of Animal Reproduction
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• v.21 no.2
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• pp.167-176
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• 1997

This study was carried out to evaluate the potential for preselection of transgenic embryos prior to transfer into recipient animals. In these experiments, I used a 3.2 kb transgene which contained the neomycin resistance gene (neo) and lac Z gene driven by the $\beta$ actin promoter (iacZ Ineo). Oocytes were aspirated from abattoir ovaries, matured in TCM-199 supplemented with 10% fetal bovine serum (FBS), 5 ${\mu}\textrm{g}$/ml LH, 0.5 ${\mu}\textrm{g}$/ml FSH, 100 unit/ml penicillin, and 100 ${\mu}\textrm{g}$/ml streptomycin for 22 to 24 hrs then inseminated with a sperm suspension of 1 X 10$^6$ sperm/ml containing 5 ${\mu}\textrm{g}$/ml of heparin. At 18-20 hrs after insemination, cumulus cells were removed by vortexing and pronuclei of centrifuged zygotes microinjected with the lacZ/neo construct (3 ng/$\mu$l). All cultures were carried out in CR1aa with transfected BRL monolayers containing 3 mg/ml BSA, 20 $\mu$/ml NEM amino acids and 40 $\mu$I/ml BME amino acids. To identify the appropriate concentration of G418 for selection, non-microinjected zygotes were cultured in the presence of 0, 50, 100 and 200 $\mu$g/ml of G418. After 8 days of culture in these treatments, 44/145 (30.3%), 13/150 (8. 7%), 1/151 (0.7%) and 0/134 (0.0%) devel-oped to the blastocyst stage in 0, 50, 100 and 200 $\mu$g/ml of G418, respectively. A total of 1,127 zygotes were microinjected and placed into culture (without G418) and subsequently 710 (63.0%) cleaved. At 48 hrs post-injection, embryos ($\geq$2-cell) were randomly assigned to control (medium alone) or G418 (100 ${\mu}\textrm{g}$/ml) treatments. A control culture of 740 non-microinjected embryos from the same replicates of embryos were also placed into control medium. After 8 days in culture, 54/343 (15.7%) and 22/367 (6.0 %) of the microinjected embryos developed to the blastocyst stage in control and G418 media, respectively. A total of 151/740 (27.2%) of the non-microinjected embryos placed in the control medium developed to the blastocyst stage. The blastocysts in the control treatment had a mean of 70.7 ${\pm}$ 4.7 cells of which 23.1 ${\pm}$ 2.6 (32.7%) stained for $\beta$-Gal activity. B1astocysts in the G418 treatment had a mean of 48.8${\pm}$7.5 cells of which 40.3 ${\pm}$ 4.1 (82.6%) stained for $\beta$-Gal ac tivity (P<0.05). In the control treatment 26 of 30 (87.0%) blastocysts had some cells with $\beta$-Gal activity while all of the blastocysts in the G418 treatment had some cell with $\beta$-Gal ac tivity (P<0.05). However, ICM colonies in either control or G418 treatments were not expressed any epiblast cell except of trophetoderm celIs. The $\beta$-actin promoter/lacZ gene may not be e expression or silence expression in epiblast cells These results clearly show an enrichment of blastocysts expressing the transgene in the majority of their cells after culture in the presence of G418. The exogeneous gene was not express a and silence in ICM colonies especiallly epiblast cells except of trophectederm cells. Even though the higher rate cell number expressed of exogeneous gene in the G418 treatments, a total cell number was decrease significantly (P<0.05) of which might be also drop of the establishment of ES like-cell colonies and production of transgenic animals. However, futher studies need to determine the viability of these selected embryos and the avability of production of transgenic offspring.

• The Korean Journal of Nuclear Medicine Technology
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• v.19 no.2
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• pp.87-92
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• 2015

Purpose When the patients takes myocardial perfusion SPECT using $^{201}Tl$, the operator gives the patients an injection of $^{201}Tl$. But the uniformity correction map in SPECT uses $^{99m}Tc$ uniformity correction map. Thus, we want to compare the image quality when it uses $^{99m}Tc$ uniformity correction map and when it uses $^{201}Tl$ uniformity correction map. Materials and Methods Phantom study is performed. We take the data by Asan medical center daily QC condition with flood phantom including $^{201}Tl$ 21.3 kBq/mL. After postprocessing with this data, we analyze CFOV integral uniformity(I.U) and differential uniformity(D.U). And we take the data with Jaszczak ECT Phantom by American college of radiology accreditation program instruction including $^{201}Tl$ 33.4 kBq/mL. After post processing with this data, we analyze spatial Resolution, Integral Uniformity(I.U), coefficient of variation(C.V) and Contrast with Interactive data language program. Results In the flood phantom test, when it uses $^{99m}Tc$ uniformity correction map, Flood I.U is 3.6% and D.U is 3.0%. When it uses $^{201}Tl$ uniformity correction map, Flood I.U is 3.8% and D.U is 2.1%. The flood I.U is worsen about 5%, but the D.U is improved about 30% inversely. In the Jaszczak ECT phantom test, when it uses $^{99m}Tc$ uniformity correction map, SPECT I.U, C.V and contrast is 13.99%, 4.89% and 0.69. When it uses $^{201}Tl$ uniformity correction map, SPECT I.U, C.V and contrast is 11.37%, 4.79% and 0.78. All of data are improved about 18%, 2%, 13% The spatial resolution was no significant changes. Conclusion In the flood phantom test, Flood I.U is worsen but Flood D.U is improved. Therefore, it's uncertain that an image quality is improved with flood phantom test. On the other hand, SPECT I.U, C.V, Contrast are improved about 18%, 2%, 13% in the Jaszczak ECT phantom test. This study has limitations that we can't take all variables into account and study with two phantoms. We need think about things that it has a good effect when doctors decipher the nuclear medicine image and it's possible to improve the image quality using the uniformity correction map of other radionuclides other than $^{99m}Tc$, $^{201}Tl$ when we make other nuclear medicine examinations.

• The Korean Journal of Nuclear Medicine Technology
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• v.18 no.2
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• pp.8-16
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• 2014

Purpose The quantitative analysis of gallbladder emptying is very important in diagnosis of motility disorder of gallbladder and in biliary physiology. The GBEF obtain the statics aquisition method or the dynamic acquisition method in two ways. The purpose of this study is to compare the GBEF value of statics acquisition method and the dynamic acquisition method. And we find the best way for calculate GBEF. Materials and Methods The quantitative hepatobiliary scan with $^{99m}Tc$-mebrofenin was performed of 27 patients. Initial images were acquired statically, for 60 min after injection of the radioactive tracer. And if the gallbladder is visualized to 60 min, performed stimulation of gallbladder (1egg, 200 mL milk). After that, started acquisition of dynamic image for 30 min. After that, image of after fatty meal of the statics method were acquired on equal terms with 60 min image. The statics GBEF was calculated using the images of before fatty meal and post fatty meal by the statics method. The dynamic GBEF was calculated using the images of time of maximum bile juice uptake ($T_{max}$) and time of minimum bile juice uptake ($T_{min}$) images from the gallbladder time-activity curve. A bile juice is secreted from gallbladder while eating a fatty meal. that is named early GBEF and that was calculated using before fatty meal image of the statics method and 1 min image of the dynamic method. Results The result saw very big difference between two according to $T_{max}$. The result, were as follows. 1) In case of less than 1 min, the dynamic mean GBEF was $40.1{\pm}21.7%$, the statics mean GBEF was $51.5{\pm}23.6%$ in 16 cases. The early mean GBEF was $14.0{\pm}29.1%$. The GBEF of statics method was higher because that include secreted bile juice while performed stimulation of gallbladder. A difference of GB counts according to acquisition method and the early bile juice counts was $17.6{\pm}14.8%$ and $13.5{\pm}15.3%$. 2) In case of exceed than 1 min, the dynamic mean GBEF was $31.0{\pm}19.7%$, the statics mean GBEF was $21.3{\pm}19.4%$ in 7 cases. The early GBEF was $-6.9{\pm}4.9%$. The GBEF of dynamic method was higher because that include concentrated bile juice to $T_{max}$. A difference of GB counts according to acquisition method and the early bile juice counts was $14.3{\pm}7.3%$ and $5.9{\pm}3.9%$. Conclusion The statics method is very easy and simple, but in case of $T_{max}$ delay, the GBEF can be lower. The dynamic method is able to calculate accurately in case of $T_{max}$ delay, but in case of $T_{max}$ is less than 1 min, the GBEF can be lower because dynamic GBEF exclude secreted bile juice while performed stimulation of gallbladder. The best way to calculate GBEF is to scan with dynamic method preferentially and to choose suitable method between the two way after conform $T_{max}$ on the T-A curve of the dynamic method.

• Progress in Medical Physics
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• v.22 no.3
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• pp.140-147
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• 2011

The purpose of this study was to estimate internal motion using molecular sieve for quantitative improvement of lung tumor and to localize lung tumor in the small animal PET image by evaluated data. Internal motion has been demonstrated in small animal lung region by molecular sieve contained radioactive substance. Molecular sieve for internal lung motion target was contained approximately 37 kBq Cu-64. The small animal PET images were obtained from Siemens Inveon scanner using external trigger system (BioVet). SD-Rat PET images were obtained at 60 min post injection of FDG 37 MBq/0.2 mL via tail vein for 20 min. Each line of response in the list-mode data was converted to sinogram gated frames (2~16 bin) by trigger signal obtained from BioVet. The sinogram data was reconstructed using OSEM 2D with 4 iterations. PET images were evaluated with count, SNR, FWHM from ROI drawn in the target region for quantitative tumor analysis. The size of molecular sieve motion target was $1.59{\times}2.50mm$. The reference motion target FWHM of vertical and horizontal was 2.91 mm and 1.43 mm, respectively. The vertical FWHM of static, 4 bin and 8 bin was 3.90 mm, 3.74 mm, and 3.16 mm, respectively. The horizontal FWHM of static, 4 bin and 8 bin was 2.21 mm, 2.06 mm, and 1.60 mm, respectively. Count of static, 4 bin, 8 bin, 12 bin and 16 bin was 4.10, 4.83, 5.59, 5.38, and 5.31, respectively. The SNR of static, 4 bin, 8 bin, 12 bin and 16 bin was 4.18, 4.05, 4.22, 3.89, and 3.58, respectively. The FWHM were improved in accordance with gate number increase. The count and SNR were not proportionately improve with gate number, but shown the highest value in specific bin number. We measured the optimal gate number what minimize the SNR loss and gain improved count when imaging lung tumor in small animal. The internal motion estimation provide localized tumor image and will be a useful method for organ motion prediction modeling without external motion monitoring system.

• Nuclear Medicine and Molecular Imaging
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• v.43 no.1
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• pp.10-18
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• 2009

Purpose: We investigated quantification of dopaminergic transporter (DAT) and serotonergic transporter (SERT) on $^{123}I$-FP-CIT SPECT for differentiating between multiple systemic atrophy (MSA) and idiopathic Parkinson's disease (IPD). Materials and Methods: N-fluoropropyl-$2{\beta}$-carbomethoxy-$3{\beta}$-4-[$^{123}I$]-iodophenylnortropane SPECT ($^{123}I$-FP-CIT SPECT) was performed in 8 patients with MSA (mean age: $64.0{\pm}4.5yrs$, m:f=6:2), 13 with early IPD (mean age: $65.5{\pm}5.3yrs$, m:f=9:4), and 12 healthy controls (mean age: $63.3{\pm}5.7yrs$, m:f=8:4). Standard regions of interests (ROls) of striatum to evaluate DAT, and hypothalamus and midbrain for SERT were drawn on standard template images and applied to each image taken 4 hours after radiotracer injection. Striatal specific binding for DAT and hypothalamic and midbrain specific binding for SERT were calculated using region/reference ratio based on the transient equilibrium method. Group differences were tested using ANOVA with the postHoc analysis. Results: DAT in the whole striatum and striatal subregions were significantly decreased in both patient groups with MSA and early IPD, compared with healthy control (p<0.05 in all). In early IPD, a significant increase in the uptake ratio in anterior and posterior putamen and a trend of increase in caudate to putamen ratio was observed. In MSA, the decrease of DAT was accompanied with no difference in the striatal uptake pattern compared with healthy controls. Regarding the brain regions where $^{123}I$-FP-CIT binding was predominant by SERT, MSA patients showed a decrease in the binding of $^{123}I$-FP-CIT in the pons compared with controls as well as early IPD patients (MSA: $0.22{\pm}0.1$ healthy controls: $0.33{\pm}0.19$, IPD: $0.29{\pm}0.19$), however, it did not reach the statistical significance. Conclusion: In this study, the differential patterns in the reduction of DAT in the striatum and the reduction of pontine $^{123}I$-FP-CIT binding predominant by SERT could be observed in MSA patients on $^{123}I$-FP-CIT SPECT. We suggest that the quantification of SERT as well as DAT using $^{123}I$-FP-CIT SPECT is helpful to differentiate parkinsonian disorders in early stage.