• Progress in Medical Physics
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• v.20 no.4
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• pp.269-276
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• 2009

Radiation treatment techniques using photon beam such as three-dimensional conformal radiation therapy (3D-CRT) as well as intensity modulated radiotherapy treatment (IMRT) demand accurate dose calculation in order to increase target coverage and spare healthy tissue. Both jaw collimator and multi-leaf collimators (MLCs) for photon beams have been used to achieve such goals. In the Pinnacle3 treatment planning system (TPS), which we are using in our clinics, a set of model parameters like jaw collimator transmission factor (JTF) and MLC transmission factor (MLCTF) are determined from the measured data because it is using a model-based photon dose algorithm. However, model parameters obtained by this auto-modeling process can be different from those by direct measurement, which can have a dosimetric effect on the dose distribution. In this paper we estimated JTF and MLCTF obtained by the auto-modeling process in the Pinnacle3 TPS. At first, we obtained JTF and MLCTF by direct measurement, which were the ratio of the output at the reference depth under the closed jaw collimator (MLCs for MLCTF) to that at the same depth with the field size $10{\times}10\;cm^2$ in the water phantom. And then JTF and MLCTF were also obtained by auto-modeling process. And we evaluated the dose difference through phantom and patient study in the 3D-CRT plan. For direct measurement, JTF was 0.001966 for 6 MV and 0.002971 for 10 MV, and MLCTF was 0.01657 for 6 MV and 0.01925 for 10 MV. On the other hand, for auto-modeling process, JTF was 0.001983 for 6 MV and 0.010431 for 10 MV, and MLCTF was 0.00188 for 6 MV and 0.00453 for 10 MV. JTF and MLCTF by direct measurement were very different from those by auto-modeling process and even more reasonable considering each beam quality of 6 MV and 10 MV. These different parameters affect the dose in the low-dose region. Since the wrong estimation of JTF and MLCTF can lead some dosimetric error, comparison of direct measurement and auto-modeling of JTF and MLCTF would be helpful during the beam commissioning.

• Journal of Life Science
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• v.24 no.2
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• pp.173-180
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• 2014

Estrogen can promote or inhibit cellular proliferation depending on tissue cell types and physiological condition and acts through the signal transduction pathways mediated primarily by estrogen receptors. This study examined the effects of fulvestrant (Ful), a well-known antagonist for the estrogen receptor, on the action of $17{\beta}$-estradiol (E2) with respect to the proliferation and apoptosis of Chinese hamster ovarian (CHO) cells. We used different concentrations of E2, Ful, and E2 plus Ful during different treatment durations. Treatment with 15-40 ${\mu}M$ E2 significantly inhibited proliferation in a time-dependent manner, although it had no influence in concentrations up to 1 ${\mu}M$. Interestingly, Ful at 10-40 ${\mu}M$ also inhibited cellular proliferation in both a concentration- and time-dependent manner. In addition, Ful enhanced rather than decreased the inhibitory effect on cellular proliferation by E2 in combined treatment for 10 days. Thus, Ful does not appear to have an antagonistic effect on estrogen's anti-proliferative action in CHO cells. In TUNEL assays to confirm DNA fragmentation by E2 and/or Ful, CHO cells treated with 20 ${\mu}M$ E2 showed a TUNEL-positive reaction in most DAPI-stained nuclei, and cells treated with either 40 ${\mu}M$ Ful or 40 ${\mu}M$ Ful plus 20 ${\mu}M$ E2 also exhibited a TUNEL-positive reaction but at a lower rate compared to the E2-treated cells. These results indicate that Ful does not have an antagonistic effect on estrogen's anti-proliferative action in CHO cells, suggesting that the anti-proliferative and apoptosis-related mechanism(s) through DNA fragmentation by E2 and Ful may be mediated by different signal transduction pathways.

• Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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• 2001.06a
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• pp.1031-1031
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• 2001

The mammary gland is made up of remarkably sensitive tissue, which has the capability of producing a large volume of secretion, milk, under normal or healthy conditions. When bacteria enter the gland and establish an infection (mastitis), inflammation is initiated accompanied by an influx of white cells from the blood stream, by altered secretory function, and changes in the volume and composition of secretion. Cell numbers in milk are closely associated with inflammation and udder health. These somatic cell counts (SCC) are accepted as the international standard measurement of milk quality in dairy and for mastitis diagnosis. NIR Spectra of unhomogenized composite milk samples from 14 cows (healthy and mastitic), 7days after parturition and during the next 30 days of lactation were measured. Different multivariate analysis techniques were used to diagnose the disease at very early stage and determine how the spectral properties of milk vary with its composition and animal health. PLS model for prediction of somatic cell count (SCC) based on NIR milk spectra was made. The best accuracy of determination for the 1100-2500nm range was found using smoothed absorbance data and 10 PLS factors. The standard error of prediction for independent validation set of samples was 0.382, correlation coefficient 0.854 and the variation coefficient 7.63%. It has been found that SCC determination by NIR milk spectra was indirect and based on the related changes in milk composition. From the spectral changes, we learned that when mastitis occurred, the most significant factors that simultaneously influenced milk spectra were alteration of milk proteins and changes in ionic concentration of milk. It was consistent with the results we obtained further when applied 2DCOS. Two-dimensional correlation analysis of NIR milk spectra was done to assess the changes in milk composition, which occur when somatic cell count (SCC) levels vary. The synchronous correlation map revealed that when SCC increases, protein levels increase while water and lactose levels decrease. Results from the analysis of the asynchronous plot indicated that changes in water and fat absorptions occur before other milk components. In addition, the technique was used to assess the changes in milk during a period when SCC levels do not vary appreciably. Results indicated that milk components are in equilibrium and no appreciable change in a given component was seen with respect to another. This was found in both healthy and mastitic animals. However, milk components were found to vary with SCC content regardless of the range considered. This important finding demonstrates that 2-D correlation analysis may be used to track even subtle changes in milk composition in individual cows. To find out the right threshold for SCC when used for mastitis diagnosis at cow level, classification of milk samples was performed using soft independent modeling of class analogy (SIMCA) and different spectral data pretreatment. Two levels of SCC - 200 000 cells/$m\ell$ and 300 000 cells/$m\ell$, respectively, were set up and compared as thresholds to discriminate between healthy and mastitic cows. The best detection accuracy was found with 200 000 cells/$m\ell$ as threshold for mastitis and smoothed absorbance data: - 98% of the milk samples in the calibration set and 87% of the samples in the independent test set were correctly classified. When the spectral information was studied it was found that the successful mastitis diagnosis was based on reviling the spectral changes related to the corresponding changes in milk composition. NIRS combined with different ways of spectral data ruining can provide faster and nondestructive alternative to current methods for mastitis diagnosis and a new inside into disease understanding at molecular level.

• Clinical and Experimental Reproductive Medicine
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• v.33 no.3
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• pp.171-178
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• 2006

Objective: Human embryonic stem (ES) cells have a great potential in regenerative medicine and tissue engineering. The human ES cells could be differentiated into specific cell types by treatments of growth factors and alterations of gene expressions. However, the efficacy of guided differentiation and isolation of specific cells are still low. In this study, we characterized isolated cells from differentiated human ES cells by magnetic activated cell sorting (MACS) system using specific antibodies to cell surface markers. Methods: The undifferentiated hES cells (Miz-hESC4) were sub-cultured by mechanical isolation of colonies and embryoid bodies were spontaneously differentiated with DMEM containing 10% FBS for 2 weeks. The differentiated cells were isolated to positive and negative cells with MACS system using CD34, human epithelial antigen (HEA) and human fibroblast (HFB) antibodies, respectively. Observation of morphological changes and analysis of marker genes expression were performed during further culture of MACS isolated cells for 4 weeks. Results: Morphology of the CD34 positive cells was firstly round, and then it was changed to small polygonal shape after further culture. The HEA positive cells showed large polygonal, and the HFB positive spindle shape. In RT-PCR analysis of marker genes, the CD34 and HFB positive cells expressed endodermal and mesodermal genes, and HEA positive cells expressed ectodermal genes such as NESTIN and NF68KD. The marker genes expression pattern of CD34 positive cells changed during the extension of culture time. Conclusion: Our results showed the possibility of successful isolation of specific cells by MACS system from undirected differentiated human ES cells. Thus, MACS system and marker antibodies for specific cell types might be useful for guided differentiation and isolation of specific cells from human ES cells.

• Journal of Chest Surgery
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• v.37 no.6
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• pp.482-491
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• 2004

Background: The xenogenic or allogenic valves after in Vitro repopulation with autologous cells or in vivo repo-pulation after acellularization treatment to remove the antigenicity could used as an alternative to synthetic polymer scaffold. In the present study, we evaluated the process of repopulation by recipient cell to the acellu-larized xenograft treated with NaCl-SDS solution and grafted in the right ventricular outflow tract. Material and Method: Porcine pulmonary valved conduit were treated with. NaCl-SDS solution to make the grafts acellularized and implanted in the right ventricular outflow tract of the goats under cardiopulmonary bypass. After evaluating the functions of pulmonary valves by echocardiography, goats were sacrificed at 1 week, 1 month, 3 months, 6 months, and 12 months after implantation, respectively. After retrieving the implanted valved conduits, histopathologic examination with Hematoxylin-Eosin, Masson' trichrome staining and immunohistochemical staining was performed. Result: Among the six goats, which had been implanted with acellularized pulmonary valved conduits, five survived the expected time period. Echocardiographic examinations for pulmonary valves revealed good function except mild regurgitation and stenosis. Microscopic analysis of the leaflets showed progressive cellular in-growth, composed of fibroblasts, myofibroblasts, and endothelial cells, into the acellularized leaflets over time. Severe inflammatory respon-se was detected in early phase, though it gradually decreased afterwards. The extracellular matrices were regenerated by repopulated cells on the recellularized portion of the acellularized leaflet. Conclusion: The acellularized xenogenic pulmonary valved conuits were repopulated with fibroblasts, myofibroblasts, and endothelial cells of the recipient and extracellullar matrices were regenerated by repopulted cells 12 months after the implantation. The functional integrity of pulmonary valves was well preserved. This study showed that the acellularized porcine xenogenic valved conduits could be used as an ideal valve prosthesis with long term durability.

• Journal of Chest Surgery
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• v.41 no.5
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• pp.550-562
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• 2008

Background: We attempted to reproduce a previously reported method that is known to be effective for decellularization, and we sought to find the optimal condition for decellularization by introducing some modifications to this method. Material and Method: Porcine semilunar valves, arterial walls and pericardium were processed for decellularization with using a variety of combinations and concentrations of decellularizing agents under different conditions of temperature, osmolarity and incubation time. The degree of decellularization and the preservation of the extracellular matrix were evaluated by staining with hematoxylin and eosin and with alpha-Gal and DAPI in some of the decellularized tissues. Result: Decellularization was achieved in the specimens that were treated with sodium deoxycholate, sodium dodesyl sulfate, Triton X-100 and sodium dodesyl sulfate with Triton X-100 as single-step methods, and this was also achieved in the specimens that were treated with hypotonic solution ${\rightarrow}$ Triton X-100 ${\rightarrow}$ sodium dodesyl sulfate, sodium deoxycholate ${\rightarrow}$ hypotonic solution ${\rightarrow}$ sodium dodesyl sulfate, and hypotonic solution sodium dodesyl sulfate as multi-step methods. Conclusion: Considering the number and the amount of the chemicals that were used, the incubation time and the degree of damage to the extracellular matrix, a single-step method with sodium dodesyl sulfate and Triton X-100 and a multi-step method with hypotonic solution followed by sodium dodesyl sulfate were both relatively optimal methods for decellularization in this study.

• Asian-Australasian Journal of Animal Sciences
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• v.33 no.2
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• pp.219-229
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• 2020

Objective: Considering the physiological and clinical importance of leptin receptor (LEPR) in regulating obesity and the fact that porcine LEPR expression is not known to be controlled by lncRNAs and miRNAs, we aim to characterize this gene as a potential target of SSC-miR-323 and the lncRNA TCONS_00010987. Methods: Bioinformatics analyses revealed that lncRNA TCONS_00010987 and LEPR have SSC-miR-323-binding sites and that LEPR might be a target of lncRNA TCONS_00010987 based on cis prediction. Wild-type and mutant TCONS_00010987-target sequence fragments and wild-type and mutant LEPR 3'-UTR fragments were generated and cloned into pmiRRB-REPORT^TM-Control vectors to construct respective recombinant plasmids. HEK293T cells were co-transfected with the SSC-miR-323 mimics or a negative control with constructs harboring the corresponding binding sites and relative luciferase activities were determined. Tissue expression patterns of lncRNA TCONS_00010987, SSC-miR-323, and LEPR in Anqing six-end-white (AQ, the obese breed) and Large White (LW, the lean breed) pigs were detected by real-time quantitative polymerase chain reaction; backfat expression of LEPR protein was detected by western blotting. Results: Target gene fragments were successfully cloned, and the four recombinant vectors were constructed. Compared to the negative control, SSC-miR-323 mimics significantly inhibited luciferase activity from the wild-type TCONS_00010987-target sequence and wild-type LEPR-3'-UTR (p<0.01 for both) but not from the mutant TCONS_00010987-target sequence and mutant LEPR-3'-UTR (p>0.05 for both). Backfat expression levels of TCONS_00010987 and LEPR in AQ pigs were significantly higher than those in LW pigs (p<0.01), whereas levels of SSC-miR-323 in AQ pigs were significantly lower than those in LW pigs (p<0.05). LEPR protein levels in the backfat tissues of AQ pigs were markedly higher than those in LW pigs (p<0.01). Conclusion: LEPR is a potential target of SSC-miR-323, and TCONS_00010987 might act as a sponge for SSC-miR-323 to regulate LEPR expression.

• Journal of Radiation Protection and Research
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• v.27 no.1
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• pp.37-49
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• 2002

The accuracy of radiation dose delivery to target volume is one of the most important factors for good local control and less treatment complication. In vivo dosimetry is an essential QA procedure to confirm the radiation dose delivered to the patients. Transmission dose measurement is a useful method of in vivo dosimetry and it's advantages are non-invasiveness, simplicity and no additional efforts needed for dosimetry. In our department, in vivo dosimetry system using measurement of transmission dose was manufactured and algorithms for estimation of transmission dose were developed and tested with phantom in various conditions successfully. This system was applied in clinic to test stability, reproducibility and applicability to daily treatment and the accuracy of the algorithm. Transmission dose measurement was performed over three weeks. To test the reproducibility of this system, X-tay output was measured before daily treatment and then every hour during treatment time in reference condition(field size; $10 cm{\times} 10 cm$, 100 MU). Data of 11 patients whose pelvis were treated more than three times were analyzed. The reproducibility of the dosimetry system was acceptable with variations of measurement during each day and over 3 week period within ${\pm}2.0%$. On anterior- posterior and posterior fields, mean errors were between -5.20% and +2.20% without bone correction and between -0.62% and +3.32% with bone correction. On right and left lateral fields, mean errors were between -10.80% and +3.46% without bone correction and between -0.55% and +3.50% with bone correction. As the results, we could confirm the reproducibility and stability of our dosimetry system and its applicability in daily radiation treatment. We could also find that inhomogeneity correction for bone is essential and the estimated transmission doses are relatively accurate.

• Korean Journal of Microbiology
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• v.48 no.4
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• pp.314-318
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• 2012

Most types of collagen used for biomedical applications, such as cell therapy and tissue engineering, are derived from animal tissues. Therefore, special precautions must be taken during the production of these proteins in order to assure against the possibility of the products transmitting infectious diseases to the recipients. The ability to remove and/or inactivate known and potential viral contaminants during the manufacturing process is an ever-increasingly important parameter in assessing the safety of biomedical products. The purpose of this study was to evaluate the efficacies of the 70% ethanol treatment and pepsin treatment at pH 2.0 for the inactivation of bovine viruses during the manufacture of collagen type I from bovine hides. A variety of experimental model viruses for bovine viruses including bovine herpes virus (BHV), bovine viral diarrhea virus (BVDV), bovine parainfluenza 3 virus (BPIV-3), and bovine parvovirus (BPV), were chosen for the evaluation of viral inactivation efficacy. BHV, BVDV, BPIV-3, and BPV were effectively inactivated to undetectable levels within 1 h of 70% ethanol treatment for 24 h, with log reduction factors of ${\geq}5.58$, ${\geq}5.32$, ${\geq}5.11$, and ${\geq}3.42$, respectively. BHV, BVDV, BPIV-3, and BPV were also effectively inactivated to undetectable levels within 5 days of pepsin treatment for 14 days, with the log reduction factors of ${\geq}7.08$, ${\geq}6.60$, ${\geq}5.60$, and ${\geq}3.59$, respectively. The cumulative virus reduction factors of BHV, BVDV, BPIV-3, and BPV were ${\geq}12.66$, ${\geq}11.92$, ${\geq}10.71$, and ${\geq}7.01$. These results indicate that the production process for collagen type I from bovine hides has a sufficient virus-reducing capacity to achieve a high margin of virus safety.

• Journal of Korean Society of Forest Science
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• v.83 no.2
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• pp.191-204
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• 1994

Due to its topographic complexities and various climatical condition, Korea exhibits diverse forest types. Dominant tree species in this zone are Quercus spp., Betula spp., Zelkova spp., Fraxinus spp., Pinus densiflora, Pinus koraiensis, and Pinus thunbergii ete. Genetic conservation in forest species in Korea there are three ways ; one is in situ, other is ex situ and third is in-facility conservation. In situ conservation include that are the present status of conservation of rare and endangered flora and ecosystem, the reserved forest, the national and provincial park, and the gene pool of natural forests. Ex situ conservation means to be established the new forest from in situ forest stands, progeny and provenance test populations, seed orchard and clone banks, and gene conservation in-facility. As a tool for low temperature storage, several aspects on in vitro system were studied ; (1) establishment of in vitro cultures from juvenile and/or rejuvenated tissues, (2) induction of multiple shoots from the individual micropropagules, (3) elongation of the proliferated shoots. Studies on cold storage for short-and long-term maintenance of in vitro cultures under $4^{\circ}C$ in the refrigerator were conducted. For the cryopreservation at $-196^{\circ}C$, various factors affecting survivability of the plant materials are being examined. The necessity of gene conservation of forest trees is enlarged not only to increase the adaptability for various environments but also to gain the breeding materials in the future. For effective gene conservation of forest trees, I would like to suggest followings ; 1. Forest stands reserved for other than the gene conservation purposes such as national parks should be investigated by botanical and gene-ecological studies for selecting bio-diversity and gene conservation stands. 2. Reserved forest for gene pool should be extented both economically important tree spp. and non-economical species. 3. Reserved forest for progeny test and clone bank should be systematically investigated for the use of Ex situ forest gene conservation. 4. We have to find out a new methodology of genetic analysis determining the proper and effective size of subpopulation for in situ gene conservation. 5. We should develop a new tree breeding systems for successful gene conservation and utilization of the genetic resources. 6. New method of in-facility gene conservation using advanced genetic engineering should be developed to save time and economic resources. 7. For the conservation of species with short-life span of seed or shortage of knowledge of seed physiology, tissue culture techniques will be played a great role for gene conservation of those species. 8. It is are very useful conservation not only of genes but of genotypes which were selected already by breeding program. 9. Institutional and administrative arrangements including legistlation must be necessarily taken for gene conservation of forest trees. 10. It is national problems for conservation of forest resources which have been rapidly destroyed because of degenerating environmental condition and of inexperienced management system of bio-diversity and gene conservation. 11. In order to international cooperation for exchanging data of bio-diversity and gene conservation, we should connect to international net works as soon as possible.