• The Korean Journal of Nuclear Medicine
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• v.36 no.6
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• pp.317-324
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• 2002

Purpose: The use of statistical parametric mapping (SPM) program has increased for the analysis of brain PET and SPECT images. Montreal Neurological Institute (MNI) coordinate is used in SPM program as a standard anatomical framework. While the most researchers look up Talairach atlas to report the localization of the activations detected in SPM program, there is significant disparity between MNI templates and Talairach atlas. That disparity between Talairach and MNI coordinates makes the interpretation of SPM result time consuming, subjective and inaccurate. The purpose of this study was to develop a program to provide objective anatomical information of each x-y-z position in ICBM coordinate. Materials and Methods: Program was designed to provide the anatomical information for the given x-y-z position in MNI coordinate based on the Statistical Probabilistic Anatomical Map (SPAM) images of ICBM. When x-y-z position was given to the program, names of the anatomical structures with non-zero probability and the probabilities that the given position belongs to the structures were tabulated. The program was coded using IDL and JAVA language for 4he easy transplantation to any operating system or platform. Utility of this program was shown by comparing the results of this program to those of SPM program. Preliminary validation study was peformed by applying this program to the analysis of PET brain activation study of human memory in which the anatomical information on the activated areas are previously known. Results: Real time retrieval of probabilistic information with 1 mm spatial resolution was archived using the programs. Validation study showed the relevance of this program: probability that the activated area for memory belonged to hippocampal formation was more than 80%. Conclusion: These programs will be useful for the result interpretation of the image analysis peformed on MNI coordinate, as done in SPM program.

• Journal of Life Science
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• v.23 no.4
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• pp.471-478
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• 2013

Human adipose tissue-derived mesenchymal stem cells (hADSCs) have therapeutic potential, including the ability to self-renew and differentiate into multiple lineages. Understanding of molecular mechanisms of stem cell differentiation is important for improving the therapeutic efficacies of stem cell transplantation. In this study, we determined the role of nuclear factor of activated T cells (NFAT5) in the osteogenic differentiation of hADSCs. The down-regulation of NFAT5 expression by the transfection of a specific siRNA significantly inhibited osteogenic differentiation of hADSCs and decreased the activity of the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-${\kappa}B$) promoter without affecting their proliferation and adipogenic differentiation. The inhibition of NFAT5 expression inhibited the basal and Tumor Necrosis Factor ${\alpha}$ (TNF-${\alpha}$) induced activation of NF-${\kappa}B$, but it did not affect TNF-${\alpha}$-induced degradation of the $I{\kappa}B$ protein. These findings indicate that NFAT5 plays an important role in the osteogenic differentiation of hADSCs through the modulation of the NF-${\kappa}B$ pathway.

• Journal of Animal Science and Technology
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• v.46 no.2
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• pp.155-164
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• 2004

This involves identifying and cloning trapped genes from cultured cells carrying the gene-trap constructs and generating cloned zebrafish using these cells for functional study. Gene-trapping studies in gene-trapped cells were carried out in initial and cloned zebrafish carrying gene-trap events were successfully produced based on the nuclear transplantation technique. Two kind of retroviral gene-trap constructs were adopted. The first one(SA/GFP-TP), constructed in my laboratory, carries a GFP reporter gene containing a splicing acceptor and an internal neo gene. The second one(Neo-TP), obtained from Dr. Hicks (Hicks et al., 1997), contains a promoter-less neo gene located in the LTR sequence of a retroviral vector. The infected cells were subjected to drug selection(neomycin treatment) because the two constructs carry the neomycin resistant gene. All those cells survived the neomycin treatment should carry the proviral insertions. For Neo-TP, Isolated DNA from the neomycin-resistant fibroblast cells infected by Neo-TP, was digested with EcoR1 restriction enzyme and transformed into bacteria after ligation. This procedure led to the isolation of seven clones carrying flanking cellular DNA with a typical retroviral integration signature sequence. These clones contained genomic DNA ranging from 1kb to 7kb and sequences of 300-600 bp were obtained from each of the rescued plasmids. Database searching showed that all of them share high homology to zebrafish sequences. For fish cloning using tagged cells, initially, nucleus donors directly selected from a mixture of cells(Neo-TP cells) were used. A total of 44 embryos(3.7%) out of 1179 transplants were reached blastula stage; 8 of these embryos(0.8%) hatched and 3(0.3%) of them survived to adulthood. One out of three lived cloned zebrafish has an amplified fragment and was labeled with 32P.

• The Korean Journal of Nuclear Medicine Technology
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• v.23 no.2
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• pp.25-28
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• 2019

Purpose In patients with unusual kidney position after $^{99m}Tc-DTPA$ renal dynamic imaging study, the GFR(Glomerular Filtration Rate) values are significantly different according to the depth of the kidney. Thus, we tried to compare the difference of the GFR values between the depth measurement methods and in-vitro test. 30 adult patients who were subjected to renal study. 27 patients were in usual position and 3 patients were in unusual. $555{\pm}37MBq$ of $^{99m}Tc-DTPA$ was administrated to all patients. GE infinia gamma camera was used. GFR values were obtained in-vivo(gates method) and in-vitro(blood). The kidney depth in-vivo was calculated by three methods(tonnensen, manual, taylor). In-vitro, GFR was performed by blood test. Differences in the mean values of GFR and correlation between depth and GFR values were evaluated using the SPSS 12.0 statistical program. The GFR values for 27 patients with kidney in the usual position are as follows(1.tonnensen 2.manual 3.taylor 4.invitro); $69.3{\pm}4.2$, $88.2{\pm}5.6$, $77.8{\pm}4.3$, $82.2{\pm}5.8ml/min$. The three unusual cases are as follows, first(congenital renal anomaly): 66.4, 101.24, 69.07, 94.8 ml/min. second(transplantation kidney): 12.22, 29.99, 19.36, 23.5 ml/min. third(horseshoe kidney): 37.37, 93.54, 35.9, 92.5 ml/min. There was a difference between tonnensen and manual in the usual position of the kidney(p<0.05). There was no significant difference between the other methods. However, there was a significant difference in case of the unusual position of the kidneys. Correlation analysis between both kidney depth and GFR value shows person correlation as follows; Rt kidney: 0.298, Lt kidney: 0.322. When compared with the GFR values in-vitro test, it was useful to calculate the GFR value by measuring the kidney depth using a manual formula in the unusual position of the kidneys. GFR values and kidney depth were significantly related.

• BMB Reports
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• v.30 no.2
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• pp.106-112
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• 1997

The effect of cryopreservation on extracellular matrix was studied with the ultimate objective of permiting a prediction of the tendency of aorta conduit tissue to calcify following transplantation. Cryopreserved and fresh porcine aorta conduit tissues were extracted using guanidine-hydrochloride (Gdn-HCl) followed by sequential digestion of the tissues with collagenase, elastase, and papain. Glycosaminoglycans (GAGs) of the proteoglycans (PGs) were isolated and quantitated. Gdn-HCl extracted about 61% and 62% of the total GAG (proteoqlycan) material from cryopreserved and fresh tissues, respectively. Collagenasesolubilized proteoglycans from Gdn-HCl extracted tissue represented 20% and 13%, respectively, of the total GAGs present in cryopreserved and fresh tissues. Subsequent elastase hydrolysis of collagenase-digested tissue released about 11% of total GAGs from cryopreserved tissue and 16% from fresh tissue. The remaining 8%, from cryopreserved tissue, and 9%, from fresh tissue, of the total GAGs were obtained after using a papain hydrolysis. There was essentially no difference between fresh and cryopreserved tissues in the relative distribution of proteoglycans in the extracts and digestions except in the initial digestion step where more proteoglycans were obtained from collagenase solubilization of cryopreserved tissue than fresh tissue (p<0.05). The histologic status of the fresh and cryopreserved porcine aortic conduit did not differ markedly. The normal tissue architecture was not affected markedly by the cryopreservation procedure as neither alteration of elastic structure, fibrous proteins nor alteration of nuclear distribution or smooth muscle cell morphology was detected. Quantitative tissue mineral studies revealed that the mean calcium content of the cryopreserved aorta conduit tissue $(165{\pm}3\;{\mu}g/g\;wet\;tissue)$ was higher than that of the fresh tissue $(105{\pm}4\;{\mu}g/g\;wet\;tissue)$ $(p<0.05)$. The mean phosphorus content was $703{\pm}35\;{\mu}g$ wet tissue from cryopreserved tissue and $720{\pm}26\;{\mu}g$ wet tissue from fresh tissue. The study indicates that there is no significant alteration in the distribution of PGs in properly cryopreserved tissue, but the total calcium level appears to be increased in tissue cryopreserved by the cryopreservation process used in this study.

• Proceedings of the KSAR Conference
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• 2001.10a
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• pp.62-62
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• 2001

Development of effective activation protocols is of great importance for improving the success of cloning and subsequent transgenic. Three methods for oocyte activation, including 5μM ionomycin (5 min) alone, ionomycin＋1.9 mM 6-dimetylaminopurine (DMAP, 3 hrs) and ionomycin＋10㎍/㎖ cycloheximide(CHX, 3 hrs) were compared for their effects of pronuclei(PN) formation, development, developmental velocity and ploidy of parthenotes to IVF control in bovine. In group of ionomycin＋DMAP, the oocytes having more 3 PN were significantly(P〈0.05) higher than in groups of ionomycin alone and of ionomycin＋CHX (45.5% vs. 0 and 0%, respectively). Activation with the ionomycin alone, ionomycin＋DMAP and ionomycin＋CHX resulted in cleavage rates of 30, 85.5 and 57.9%, respectively. The blastocysts rate of parthenotes activated by ionomycin＋DMAP treatment was significantly higher (12.3%, P〈0.05) than those of other treated groups. Chromosome analysis shows that ionomycin＋DMAP treatment greatly increases the incidence of chromosomal abnormality of the parthenotes. When compared the developmental velocity at 24 hrs after insemination and activation, 27% eggs in IVF control and 55% in DMAP treatment out of total cleaved eggs developed to 2-cell stage, respectively. Developmental velocity of parthenotes activated by ionomycin ＋DMAP treatment was significantly (P〈0.05) faster than others. From the results, we may conclude that DMAP treatment to the oocytes accelerates developmental velocity resulting in both the higher incidence of chromosome abnormality and of PN formation suggesting that CHX combined with ionomycin is suitable DMAP for the purpose of successful nuclear transplantation.

• Reproductive and Developmental Biology
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• v.28 no.1
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• pp.71-76
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• 2004

Gene delivery is one of the keen interests in animal industry as well as research on gene functions. Some of the in vivo gene delivery techniques have been successively used in various tissues for the gene therapy and transgenesis. Despite intensive efforts, it still remains to overcome problems of limited local and regional administration and low transgene expression. To improve the efficiency of gene delivery, a new procedure was tested. We injected exogenous DNA containing LacZ into the female or male gonads and then pulsed electric field. Electroporated gonads showed positive X-gal staining in many seminiferous tubules of the porcine fetal gonads. Exogenously introduced LacZ genes were also expressed in female porcine gonad. In addition, we demonstrated efficient gene delivery in gonad of adult mouse. Furthermore, we succeed to generate genetically modified germline cells showing GFP and positive X-gal signals. The results suggest that the newly developed gene delivery is an effective way of in vivo transfection in mammals. The developed gene delivery procedure should be useful in producing transgenic animals when combined with primary cell culture and nuclear transplantation.

• The Korean Journal of Nuclear Medicine
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• v.29 no.1
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• pp.79-83
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• 1995

Purpose : We analysed $^{99m}Tc-MAG_3$ renal scans to evaluate renal function of transplanted kidney and to detect various renal transplant complications, measuring the ratio of renal radioactivity at three minutes to that at 20 minutes(elimination index). Material and Methods : The fifty seven renal transplantation recipients were studied. There were 50 normal functioning transplanted kidneys as group I and 7 abnormal function-ing transplanted kidney, including 5 cases of acute renal rejection, 2 cases of acute tubular necrosis as group IIl. The protocol consisted of: (1) $^{99m}Tc-MAG_3$ 740MBq injection intravenously : (2) sequential imaging for 2min(60two-second images) followed by 30min(30 sixty-second images) : (3) drawing of region of interest(ROI) on renal imaging; (4) time-activity corves were generated from renal ROI after background subtraction, and time of maximum activity($T_{max}$) and half time of maximal peak radioactivity($T_{1/2}$) were produced in the renogram curve. (5) EI through Bischof-Delaloye method as determined on the renogram curve. Results : Normal group( I ) shows mean EI of 2.21(95.0% Confidence limit of 2.01-2.87), $T_{max}$ of 154 sec, $T_{1/2}$ of 1,139 sec. Abnormal group(II) shows mean EI of 0.74, $T_{max}$ of 1,466 sec, $T_{1/2}$ of 19,224 sec. The EI, $T_{max}$, $T_{1/2}$, BUN and serum creatinine values are significantly different between normal group(I) and abnormal group(II) (p<0.0001). Conclusion : By measuring EI with $^{99m}Tc-MAG_3$, renal function of transplanted kidney could be easily evaluated and various complications could be detected early.

• The Korean Journal of Nuclear Medicine
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• v.36 no.4
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• pp.271-276
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• 2002

Purpose: To measure reliable glomerular filtration rate by using the representative values of transplanted renal depths, which are measured with ultrasonography. Materials and Methods: We included 54 patients (26 men, 28 women), with having both renal scintigraphy and ultrasonography after renal transplantation. We measured GFR with Gates' method using the renal depth measured by ultrasonography, and median and mean ones in each patient. We compared GFR derived from ultrasonography-measured renal depth with GFR derived from median and mean renal depths. The correlation coefficients were obtained among GFR derived from ultrasonography-measured renal depths, median and mean renal depths under linear regression analysis. We determined whether GFR derived from median or mean renal depth could substitute GFR derived from ultrasonography-measured renal depth with Bland-Altman method. We analyze the expected errors of the GFR using representative renal depth in terms of age, sex, weight, height, creatinine value, and body surface. Results: The transplanted renal depths range from 3.20 cm to 5.96 cm. The mean value and standard deviation of renal depths measured by ultrasonography are $4.09{\pm}0.65cm$ in men, and $4.24{\pm}0.78cm$ in women. The median value of renal depths measured by ultrasonography is 4.36 cm in men and 4.14 cm in women. The GFR derived from median renal depth is more consistent with GFR derived from ultrasonography-measured renal depth than GFR derived from mean renal depth. Differences of GFR derived from median and ultrasonography-measured renal depth are not significantly different in the groups classified with creatinine value, age, sex, height, weight and body surface. Conclusion: When median value is adapted as a representative renal depth, we could obtain reliable GFR in transplanted kidney simply.

• The Korean Journal of Nuclear Medicine
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• v.33 no.6
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• pp.519-526
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• 1999

Purpose: This study was designed to evaluate the usefulness of a technetium-99m mercaptoacetyltriglycine (Tc-99m MAG3) single photon emission computed tomography (SPECT) performed on transplanted kidney. Materials and Methods: Thirty renal transplant patients were included in this study. Planar scan was performed for 30 minutes using 555 MBq Tc-99m MAG3. A post-voiding SPECT scan was acquired on the third, seventh, fourteenth and twenty eighth day after transplantation. Results: SPECT scan showed interpretable image quality in 26 of 30 patients (86.7%) and 84 in 120 scans (70%). Fourteen of 26 patients with interpretable SPECT image showed decreased or increased radioactivity, but only 5 had abnormal findings on the planar scan. Focal SPECT defects were seen in allografts with normal function (n=3), acute tubular necrosis (n=3), and acute rejection (n=2). The defects are thought to reflect focally underperfused renal parenchyme or, in normal allografts, an artifact from uneven radioactivity distribution. Four of 10 patients with renal arterial variation showed focally decreased radioactivity and SPECT helped guide funker studies that confirmed the exact cause. Five of 10 patients with acute tubular necrosis or acute rejection showed focally decreased radioactivity, but its relation to the patients' clinical course was not clear. Focally increased radioactivity was observed in 5 allografts with normal function and 1 with double ureter in which local clearance delay was observed. Conclusion: Tc-99m MAG3 SPECT renal scan can detect additional focal abnormalities compared to planar scan. Further study is necessary to elucidate the exact clinical significance of the SPECT findings.