• Applied Microscopy
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• v.35 no.1
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• pp.31-39
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• 2005

Synapses are contact points where one neuron communicates with another. The morphological change of synapses under various physiological or pathological conditions has long been hypothesized to modify their functional properties. 3-dimensional (3-D) reconstruction of synapses with serial ultrathin sections has contributed to the understanding of ultrastructural dimensions and compositions of synapses. The 3-D reconstruction procedures, however, require a great amount of expertise as well as include prohibitively timeconsuming processes. Here, we introduce efficient 3-D reconstruction technique using high-voltage electron microscopy (HVEM). Primarily, we established an optimal section thickness and staining condition to observe synaptic structures in detail under HVEM. The result showed that synaptic profiles were preserved at the section thickness of 250 nm without the overlapping of synaptic ultrastructures. An increase in the reaction time of en bloc staining was most efficient to enhance contrast than the extension of postembedding staining or the addition of uranyl acetate during dehydration. Then, 3-D reconstruction of parallel fiber-Purkinje cell synapses in the rat cerebellum was carried out with serial HVEM images and reconstruction software. The images were aligned and the contours of synapses were outlined on each section. 3-D synapses were finally extracted from the section files by grouping all the synaptic contours. The reconstructed synapse model clearly demonstrated the configuration of pre and postsynaptic components. These results suggest that 3-D reconstruction of synapses using HVEM is much efficient and suitable for massive quantitative studies on synaptic connectivity than conventional TEM approach using numerous ultrathin sections.

• Korean Journal of Microbiology
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• v.52 no.4
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• pp.406-414
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• 2016

Among 6 leu codons, CUG is the most frequently used codon in E. coli. It is recognized by leu-tRNA(CAG) encoded by four genes scattered on two chromosomal loci (leuT and leuPQV ). In the process of constructing a strain with no functional leu-tRNA (CAG) gene on chromosome, we made two mutant strains separately, one on leuPQV locus (${\Delta}leuPQV$), and the other on leuT locus [$leuT^*$(GAG)], where the anticodon of leuT was changed from CAG to GAG, thereby altering its recognition codon from CUG to CUC. We attempted to combine these two mutations by transduction using $leuT^*$(GAG) strain as a donor and ${\Delta}leuPQV$ strain as a recipient. Large and small colonies appeared from this transduction. From PCR and DNA sequencing, large colony was confirmed to be the reciprocal recombinant as expected, but the small colonies contained both mutant $leuT^*$(GAG) and wild type leuT (CAG) genes in the cell. This heterozygous diploid strain did not show any unusual morphology under microscopic observation, but, interestingly, it showed a linear growth curve in rich medium with much slower growth rate than wild type cell. It always formed homogenous small colonies in the selection medium, but, when there was no selection, it readily segregated into $leuT^*$(GAG) and leuT (CAG). From these observations, we suggested that the strain with both $leuT^*$(GAG) and leuT (CAG) genes was not a partial diploid (merodiploid), but a full diploid cell having two different chromosomes. We proposed a model explaining how such a heterozygous diploid cell was formed and how and why its growth showed a linear growth curve.

• Journal of Genetic Medicine
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• v.5 no.1
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• pp.34-40
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• 2008

Purpose : Preeclampsia is a major cause of maternal and perinatal mortality and morbidity and is considered to be a multifactorial disorder involving a genetic predisposition and environmental factors. Endothelin-1 (ET-1) is a potent vasoconstrictor peptide, and alterations in the ET-1 system are thought to play a role in triggering the vasoconstriction seen with preeclampsia. The aim of this study was to examine the frequency of the 4 common single-nucleotide polymorphisms (SNPs) (c.1370T>G, c.137_139delinsA, c.3539+2T>C, and c.5665G>T) of the ET-1 gene in normotensive and preeclamptic pregnancies and to investigate whether these SNPs are associated with preeclampsia in pregnant Korean women. Methods : We analyzed blood samples from 206 preeclamptic and 216 normotensive pregnancies using a commercially available SNapShot kit and an ABI Prism 3100 Genetic analyzer. Results : There were no significant differences in genotype or allele frequencies of the 4 SNPs in the ET-1 gene between preeclamptic and normotensive pregnancies. The respective frequencies of the 3 haplotypes (TDTG, GDCT, and TICT; >10% haplotype frequency) were 61%, 13% and 13%, respectively, in preeclampsic pregnancies and 62%, 14% and 12%, respectively, in normotensive pregnancies. The frequencies of these haplotypes were similar for both groups. Using multiple logistic regression analysis, we did not observe an increase in the risk of preeclampsia for the 4 SNPs of the ET-1 gene under either a recessive or dominant model. Conclusion : This study suggests that the 4 SNPs of the ET-1 gene are not associated with an increased risk for preeclampsia in pregnant Korean women.

• 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 the Korean Society of Radiology
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• v.17 no.2
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• pp.257-265
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• 2023

The purpose of this study was to vaccinate every year according to the general characteristics of COVID-19, whether to vaccinate every year according to the vaccination experience, whether to vaccinate every year according to knowledge/attitude about vaccination, and negative responses to the vaccinate every year In order to understand the factors affecting the vaccination physician every year by identifying the factors of Statistical analysis is based on general characteristics, variables based on vaccination experience, and knowledge/attitudes related to vaccination. The doctor calculates the frequency and percentage, A square test (-test) was performed, and if the chi-square test was significant but the expected frequency was less than 5 for 25% or more, a ratio difference test was performed with Fisher's exact test. Through multiple logistic regression analysis using variables that were significant in simple analysis, a predictive model for future vaccination and the effect size of each independent variable were estimated. As statistical analysis software, SAS 9.4 (SAS Institute Inc., Cary, NC, USA) was used, and because the sample size was not large, the significance level was set at 10%, and when the p-value was less than 0.10, it was interpreted as statistically significant. In the simple logistic regression analysis, the reason why they answered that they would not be vaccinated every year was that they answered 'to prevent infection of family and hospital guests' rather than 'to prevent my infection' as the reason for the vaccination. It was 11.0 times higher and 3.67 times higher in the case of 'for the formation of collective immunity of the local community and the country'. The adverse reactions experienced after the 1st and 2nd vaccination were 8.42 times higher in those who did not experience pain at the injection site than those who did not, 4.00 times higher in those who experienced swelling or redness, and 5.69 times higher in those who experienced joint pain. There was a 5.57 times higher rate of absenteeism annually than those who did not. In addition, the more anxious they felt about vaccination, the more likely they were to not get the vaccine every year by 2.94 times.

• Health Policy and Management
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• v.34 no.3
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• pp.271-281
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• 2024

Background: The unmet healthcare needs (UHNs) of people with disabilities (PWD) are not only detrimental to their quality of life but also can lead to serious health outcomes including death. A variety of factors including socioeconomic, personal, and environmental factors affect UHNs for PWD. Previous studies focused on individual socioeconomic and disability characteristics as influencing factors. Our studies included environmental factors that have a significant impact on the use of healthcare service by PWD. Methods: We analyzed the UHNs status and influencing factors among 4,326 adults with disabilities using the Korea Disability Life Data. Chisquare analysis identified differences in UHNs by general, disability, and environmental characteristics. Logistic regression determined factors affecting UHNs. Results: Those with low educational level (adjusted odds ratio [aOR], 1.229; 95% confidence interval [CI], 1.024-1.475), those with low income level (aOR, 1.416; 95% CI, 1.015-1.976), those who enrolled in private insurance (aOR, 1.234; 95% CI, 1.018-1.496), those who need help with daily living (aOR, 1.298; 95% CI, 1.059-1.592), those who did not go out (OR, 1.566; 95% CI, 1.274-1.924), those who use taxis (aOR, 1.407; 95% CI, 1.047-1.891) or call taxi for people with disabilities when going to the hospital (aOR, 1.370; 95% CI, 1.001-1.875), the communication disabled (aOR, 1.304; 95% CI, 1.029-1.651), those with poor subjective health status (aOR, 1.248; 95% CI, 1.043-1.494), those who felt the explanation of treatment results was insufficient (aOR, 4.035; 95% CI, 1.365-11.927), hose dissatisfied with healthcare services (aOR, 3.515; 95% CI, 2.741-4.508) were more likely to experience UHNs. Conclusion: Effective healthcare service provision for PWD requires not only financial assistance but also social support, along with education for healthcare staff, policies that consider the characteristics of disabilities.

• Journal of Chest Surgery
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• v.37 no.3
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• pp.201-209
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• 2004

Extracorporeal life support (ECLS) system is a device for respiratory and/or heart failure treatment, and there have been many trials for development and clinical application in the world. Currently, a non-pulsatile blood pump is a standard for ECLS system. Although a pulsatile blood pump is advantageous in physiologic aspects, high pressure generated in the circuits and resultant blood cell trauma remain major concerns which make one reluctant to use a pulsatile blood pump in artificial lung circuits containing a membrane oxygenator. The study was designed to evaluate the hypothesis that placement of a pressure-relieving compliance chamber between a pulsatile pump and a membrane oxygenator might reduce the above mentioned side effects while providing physiologic pulsatile blood flow. The study was performed in a canine model of oleic acid induced acute lung injury (N=16). The animals were divided into three groups according to the type of pump used and the presence of the compliance chamber, In group 1, a non-pulsatile centrifugal pump was used as a control (n=6). In group 2 (n=4), a single-pulsatile pump was used. In group 3 (n=6), a single-pulsatile pump equipped with a compliance chamber was used. The experimental model was a partial bypass between the right atrium and the aorta at a pump flow of 1.8∼2 L/min for 2 hours. The observed parameters were focused on hemodynamic changes, intra-circuit pressure, laboratory studies for blood profile, and the effect on blood cell trauma. In hemodynamics, the pulsatile group II & III generated higher arterial pulse pressure (47$\pm$ 10 and 41 $\pm$ 9 mmHg) than the nonpulsatile group 1 (17 $\pm$ 7 mmHg, p<0.001). The intra-circuit pressure at membrane oxygenator were 222 $\pm$ 8 mmHg in group 1, 739 $\pm$ 35 mmHg in group 2, and 470 $\pm$ 17 mmHg in group 3 (p<0.001). At 2 hour bypass, arterial oxygen partial pressures were significantly higher in the pulsatile group 2 & 3 than in the non-pulsatile group 1 (77 $\pm$ 41 mmHg in group 1, 96 $\pm$ 48 mmHg in group 2, and 97 $\pm$ 25 mmHg in group 3: p<0.05). The levels of plasma free hemoglobin which was an indicator of blood cell trauma were lowest in group 1, highest in group 2, and significantly decreased in group 3 (55.7 $\pm$ 43.3, 162.8 $\pm$ 113.6, 82.5 $\pm$ 25.1 mg%, respectively; p<0.05). Other laboratory findings for blood profile were not different. The above results imply that the pulsatile blood pump is beneficial in oxygenation while deleterious in the aspects to high pressure generation in the circuits and blood cell trauma. However, when a pressure-relieving compliance chamber is applied between the pulsatile pump and a membrane oxygenator, it can significantly reduce the high circuit pressure and result in low blood cell trauma.

• Progress in Medical Physics
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• v.25 no.2
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• pp.100-109
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• 2014

In stereotactic body radiotherapy (SBRT), the accurate location of treatment sites should be guaranteed from the respiratory motions of patients. Lots of studies on this topic have been conducted. In this letter, a new verification method simulating the real respiratory motion of heterogenous treatment regions was proposed to investigate the accuracy of lung SBRT for Volumetric Modulated Arc Therapy. Based on the CT images of lung cancer patients, lung phantoms were fabricated to equip in $QUASAR^{TM}$ respiratory moving phantom using 3D printer. The phantom was bisected in order to measure 2D dose distributions by the insertion of EBT3 film. To ensure the dose calculation accuracy in heterogeneous condition, The homogeneous plastic phantom were also utilized. Two dose algorithms; Analytical Anisotropic Algorithm (AAA) and AcurosXB (AXB) were applied in plan dose calculation processes. In order to evaluate the accuracy of treatments under respiratory motion, we analyzed the gamma index between the plan dose and film dose measured under various moving conditions; static and moving target with or without gating. The CT number of GTV region was 78 HU for real patient and 92 HU for the homemade lung phantom. The gamma pass rates with 3%/3 mm criteria between the plan dose calculated by AAA algorithm and the film doses measured in heterogeneous lung phantom under gated and no gated beam delivery with respiratory motion were 88% and 78%. In static case, 95% of gamma pass rate was presented. In the all cases of homogeneous phantom, the gamma pass rates were more than 99%. Applied AcurosXB algorithm, for heterogeneous phantom, more than 98% and for homogeneous phantom, more than 99% of gamma pass rates were achieved. Since the respiratory amplitude was relatively small and the breath pattern had the longer exhale phase than inhale, the gamma pass rates in 3%/3 mm criteria didn't make any significant difference for various motion conditions. In this study, the new phantom model of 4D dose distribution verification using patient-specific lung phantoms moving in real breathing patterns was successfully implemented. It was also evaluated that the model provides the capability to verify dose distributions delivered in the more realistic condition and also the accuracy of dose calculation.

• 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.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.1
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• pp.47-57
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• 2000

Objective: The effects of cryopreservation with or without coculture on the in vitro development of blastomere-biopsied 8-cell mouse embryos were investigated. This experimental study was originally designed for the setup of a preclinical mouse model for the preimplantation genetic diagnosis (PGD) in human. Methods: Eight-cell embryos were obtained after in vitro fertilization (IVF) from F1 hybrid mice (C57BL(표현불가)/CBA(표현불가)). Using micromanipulation, one to four blastomeres were aspirated through a hole made in the zona pellucida by zona drilling (ZD) with acid Tyrode's solution (ATS). A slow-freezing and rapid-thawing protocol with 1.5M dimethyl sulfoxide (DMSO) and 0.1M sucrose as cryoprotectant was used for the cryopreservation of blastomere- biopsied 8-cell mouse embryos. After thawing, embryos were cultured for 110 hours in Ham's F-10 supplemented with 0.4% bovine serum albumin (BSA). In the coculture group, embryos were cultured for 110 hours on the monolayer of Vero cells in the same medium. The blastocyst formation was recorded, and the embryos developed beyond blastocyst stage were stained with 10% Giemsa to count the total number of nuclei in each embryo. Results: The survival rate of embryos after cryopreservation was significantly lower in the blastomere-biopsied (7/8, 6/8, 5/8, and 4/8 embryos) groups than in the non-biopsied, zona intact (ZI) group. Without the coculture, the blastocyst formation rate of embryos after cryopreservation was not significantly different among ZI, the zona drilling only (ZD), and the balstomere-biopsied groups, but it was significantly lower than in the non-cryopreserved control group. The mean number of cells in embryos beyond blastocyst stage was significantly higher in the control group ($50.2{\pm}14.0$) than in 6/8 ($26.5{\pm}6.2$), 5/8 ($25.0{\pm}5.5$), and 4/8 ($17.8{\pm}7.8$) groups. With the coculture using Vero cells, the blastocyst formation rate of embryos after cryopreservation was significantly lower in 5/8 and 4/8 groups, compared with the control, 7/8, and 6/8 groups. The mean number of cells in embryos beyond blastocyst stage was also significantly lower in 4/8 group ($25.9{\pm}10.2$), compared with the control ($50.2{\pm}14.0$), 7/8 ($56.0{\pm}22.2$), and 6/8 ($55.3{\pm}25.5$) groups. Conclusion: After cryopreservation, blastomere-biopsied mouse embryos have a significantly impaired developmental competence in vitro, but this detrimental effect might be prevented by the coculture with Vero cells in 8-cell mouse embryos biopsied one or two blastomeres. Biopsy of mouse embryos after ZD with ATS is a safe and highly efficient preclinical model for PGD of human embryos.