• Title/Summary/Keyword: Yo-Pro-1

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Quantification of DNA Delivery Efficiency Labeled with Fluorescent Dye in Digital Electroporation System (디지털 전기천공시스템에서 형광 염료로 표지 된 DNA 전달 효율의 정량화)

  • Bae, Seo Jun;Im, Do Jin
    • Korean Chemical Engineering Research
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    • v.58 no.3
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    • pp.450-457
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    • 2020
  • In the previous study, there was a big difference between the tendency of the delivery efficiency of Yo-Pro-1 and the expression efficiency of the CFP gene, but there was a problem that could not provide a clue to this problem. Therefore, this study aimed to present a clue to this problem by quantifying and comparing the delivery efficiency after labeling DNA using a fluorescent dye, which was one of the methods for quantifying biomolecules. As a fluorescent dye for labeling, Yo-Pro-1 was used, and the delivery efficiency of the fluorescent dye Yo-Pro-1 and the labeled DNA was compared. The delivery efficiency of Yo-Pro-1 and labeled DNA according to the voltage condition of the digital electroporation system was maximized at 96 V, and the delivery efficiency tended to decrease as the voltage increased further. In the comparison of the delivery efficiency of Yo-Pro-1 and labeled DNA according to the number of voltage application conditions, the delivery efficiency was maximized at the number of 8 voltage application times for both delivery materials, and the delivery efficiency tended to decrease as the number of voltage application increases further. Through the two results, it was confirmed that the delivery efficiency using Yo-Pro-1 in the digital electroporation system represents the delivery efficiency of the system well. In addition, through the results of this study, the difference between the tendency of the delivery efficiency of Yo-Pro-1 and the expression efficiency of the CFP gene shown in the preceding study was not the result of the difference in the delivery efficiency of the delivery material, but it can be predicted to be due to a problem with the expression process of the genetic material that had been delivered.

Evaluation of Cell Viability and Delivery Efficiency in Electroporation System According to the Concentrations of Propidium Iodide and Yo-Pro-1 (전기천공시스템에서 Propidium Iodide와 Yo-Pro-1의 농도에 따른 세포 생존율과 전달효율 평가)

  • Bae, Seo Jun;Im, Do Jin
    • Korean Chemical Engineering Research
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    • v.57 no.6
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    • pp.898-906
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    • 2019
  • In this study, basic research was conducted to provide guidelines for selecting fluorescent dye and using proper concentration of fluorescent dye to use evaluation of cell viability and fluorescent dye delivery efficiency. Propidium iodide and Yo-Pro-1 were used as fluorescent dyes. In the evaluation of cell viability and the efficiency of delivery using Propidium Iodide and Yo-Pro-1, the histogram of each fluorescent dye was different depending on the type of fluorescent dye and the concentration used. These results were related to the characteristics of the fluorescent dyes used. This was related to the properties of the fluorescent dyes used. From these results, it was found that the analytical results depending on the characteristics of the fluorescent dyes used in the cell analysis. The effect of the fluorescent dye on the cell was confirmed, but it was confirmed that it did not affect the analysis result. In addition, the influence of interference between fluorescent signals was confirmed when two or more kinds of fluorescent dyes were used for analysis. The higher the concentration of Yo-Pro-1 was, the larger the effect of interference was, and the concentration of Propidium Iodide did not affect the interference of fluorescence signal. This study confirmed that the evaluation of the cell viability and the evaluation of the delivery efficiency were influenced by the type and concentration of the fluorescent dyes and it was related to the characteristics of the fluorescent dyes. Based on the results, appropriate concentrations of fluorescent dyes suitable for evaluation of cell viability and delivery efficiency were suggested.

Delivery of Protein into Microalgae by the Digital Electroporation (디지털 전기천공을 이용한 미세조류 내 단백질 전달 연구)

  • Im, Do Jin
    • Korean Chemical Engineering Research
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    • v.56 no.1
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    • pp.79-84
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    • 2018
  • In the present study, we performed electroporation to deliver protein into microalgae using previously developed digital electroporation system. Green fluorescence protein was successfully delivered into a live microalgae cell nucleus without cell wall removal. By investigating the effects of applied voltage on the protein delivery efficiency, optimal electroporation electric field condition was found (960 V/cm). We also investigated the delivery of Yo-Pro-1 into cell to examine the size effects of delivered materials and found that there is little size effects on the optimal condition. Finally, the implications of the present results and future work are discussed.

Comparative Study of Anti-inflammatory and Immunological Activities by Different Gender and Parts of Yeonsan Ogye (연산오계의 성별과 부위별 항염증 및 면역 활성 비교 연구)

  • Do, Young Min;Kim, Dong Hee
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.32 no.2
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    • pp.99-105
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    • 2018
  • The aim of this study is to compare the anti-inflammatory and immunological activity of different parts (bone, meat, and rind) of Yeonsan Ogye (YO). In order to evaluate cytotoxicity, MTT assay was performed. We investigated the production of nitric oxide (NO) and pro-inflammatory cytokines, such as IL-$1{\beta}$, IL-6, and TNF-${\alpha}$, in LPS-induced RAW264.7 cells. All parts of the YO showed no toxicity at concentrations of 1, 10, and $100{\mu}g/m{\ell}$. Rooster's bone, hen's bone, and rind decreased the production of NO. And rooster's bone, meat, and hen's bone also attenuated TNF-${\alpha}$ production in LPS-induced RAW 264.7 cells. In addition, all parts of the YO decreased IL-$1{\beta}$ and IL-6 production in LPS-induced RAW264.7 cells, whereas they all increased IL-$1{\beta}$, IL-6 and TNF-${\alpha}$ production in normal RAW264.7 cells. Rooster exhibited higher immune activation and inhibitory activity on inflammation than a hen, and among different parts of the YO, bone showed the highest activity. Our results demonstrated and compared the anti-inflammatory and immunological activity of different parts of the YO. These results suggest that YO may be developed as a raw material for new health supplement food and medicine to attenuate various symptoms related to inflammation and immunity.

Effects of 『Geum-Gwe-Yo-Ryak(金匱要略)』 Prescription for Chest Pain Including Kwaruhaebaekbanha-tang and Kwaruhaebaekpaekju-tang on Macrophage Polarization (금궤요략(金匱要略) 심통 처방 중 과루해백반하탕과 과루해백백주탕이 대식세포 극성화에 미치는 영향)

  • Son, Chang-Hyeon;Lee, Sang-Min;Yu, Ga-Ram;Lee, Seung-Jun;Lim, Dong-Woo;Kim, Hyuck;Park, Won-Hwan;Kim, Jai-Eun
    • The Journal of Korean Medicine
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    • v.40 no.2
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    • pp.51-62
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    • 2019
  • Objectives: This study was designed to evaluate the macrophages polarization of traditional Korean medicine on cardiac pain about Geum-Gwe-Yo-Ryak's two prescriptions including Kwaruhaebaekbanha-tang (KHB) and Kwaruhaebaekpaekju-tang (KHP). Materials and methods: Flow cytometry analysis was used to measure the changes in the ratio of M1 type and M2 type macrophages. Protein expression of nuclear factor-like 2 (Nrf2), heme oxygenase-1 (HO-1), inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) were measured by Western Blot, and ABCA1 and SR-B1 were detected by real time PCR (RT-PCR). Intracellular lipid accumulation was measured by Oil Red O staining (ORO staining). Results: KHB and KHP increase anti-oxidative activity related protein levels including Nrf2 and HO-1. Furthermore, KHB and KHP inhibit lipid accumulation on intracellular levels through induction of ATP binding receptor cassette subfamily A member 1 (ABCA1) and scavenging receptor class B member 1 (SR-B1), respectively. Finally, KHB and KHP also blocked pro-inflammatory mediators including tumor necrosis factor-alpha ($TNF{\alpha}$) and interleukin-6 (IL-6), iNOS and COX-2 expression. Conclusion: This study suggests that KHB and KHP potently regulate the M1/M2 macrophage polarization.

Idiopathic infantile hypercalcemia with severe nephrocalcinosis, associated with CYP24A1 mutations: a case report

  • Yoo, Jeesun;Kang, Hee Gyung;Ahn, Yo Han
    • Childhood Kidney Diseases
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    • v.26 no.1
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    • pp.63-67
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    • 2022
  • Nephrocalcinosis often occurs in infants and is caused by excessive calcium or vitamin D supplementation, neonatal primary hyperparathyroidism, and genetic disorders. Idiopathic infantile hypercalcemia (IIH), a rare cause of nephrocalcinosis, results from genetic defects in CYP24A1 or SLC34A1. Mutations in CYP24A1, which encodes 25-hydroxyvitamin D 24-hydroxylase, disrupt active vitamin D degradation. IIH clinically manifests as failure to thrive and hypercalcemia within the first year of life and usually remits spontaneously. Herein, we present a case of IIH wih CYP24A1 mutations. An 11-month-old girl visited our hospital with incidental hypercalcemia. She showed failure to thrive, and her oral intake had decreased over time since the age of 6 months. Her initial serum parathyroid hormone level was low, 25-OH vitamin D and 1,25(OH)2 vitamin D levels were normal, and renal ultrasonography showed bilateral nephrocalcinosis. Whole-exome sequencing revealed compound heterozygous variants in CYP24A1 (NM_000782.4:c.376C>T [p.Pro126Ser] and c.1310C>A [p.Pro437His]). Although her hypercalcemia and poor oral intake spontaneously resolved in approximately 8 months, we suggested that her nephrocalcinosis and renal function be regularly checked in consideration of potential asymptomatic renal damage. Hypercalcemia caused by IIH should be suspected in infants with severe nephrocalcinosis, especially when presenting with failure to thrive.

An atypical case involving real, ghost, and pseudo-ghost images on a panoramic radiograph

  • Jong-Won Kim;Yo-Seob Seo
    • Imaging Science in Dentistry
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    • v.54 no.1
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    • pp.57-62
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    • 2024
  • Purpose: This report presents a unique case featuring real, ghost, and pseudo-ghost images on the panoramic radiograph of a patient wearing earrings. It also explains the formation of these images in an easy-to-understand manner. Materials and Methods: One real image and two ghost images appeared on each side of a panoramic radiograph of a patient wearing earrings on both sides. Of the two ghost images on each side, one was considered a typical ghost image and the other was considered a ghost-like real image (pseudo-ghost image). The formation zones of the real, double, and ghost images were examined based on the path and angles of the X-ray beam from the Planmeca ProMax. To simulate the pseudo-ghost and typical ghost images on panoramic radiography, a radiopaque marker was affixed to the right mandibular condyle of a dry mandible, and the position of the mandible was adjusted accordingly. Results: The center of rotation of the Planmeca ProMax extended beyond the jaw area, and the area of double image formation also reached beyond the jaw. The radiopaque-marked mandibular condyle, situated in the outwardly extending area of double image formation, exhibited triple images consisting of real, double (pseudo-ghost), and ghost images. These findings helped to explain the image formation associated with the patient's earrings observed in the panoramic radiograph. Conclusion: Dentists must understand the characteristics and principles of the panoramic equipment they use and apply this understanding to taking and interpreting panoramic radiographs.

Developmental Efficiency of Bovine Embryos Cloned with Fetal Fibroblast Arrested at G0/G1 Phase

  • Cho, S.R.;Son, W.J.;Park, C.S.;Park, G.J.;S.Y. Choe
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.140-140
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    • 2003
  • The study evaluated the effect of donor cell treatments for G0/Gl synchronization and the donor ceil type on development and incidence of apoptosis in cloned cattle embryos. Primary cultures were established from a female fetus on day 50 of gestation and adult ear skin biopsies. Cells were randomly allocated into 3 experimental treatment groups after 6~8 passages. Group 1 (Confluent), cells were cultured in DMEM supplemented with 10% FBS until 90% confluent. Group 2 (Serum-starvation), cells were cultured in DMEM Supplemented With 0.5% FBS for 5 days. Group 3 (Roscovitine), Cells were cultured in DMEM supplemented with 10% FBS and 30 $\mu$M Roscovitine for 12 h. Cell cycle and apoptosis were analyzed using flow cytometry after labelling with DAPI and YO-PRO-1. At 19 h post-maturation (hpm), enucleated oocytes were reconstructed with donor cells and fused by a single DC pulse (1.6 kV/cm, 60 $\mu$sec). (중략)

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Sensing Domain and Extension Rate of a Family B-Type DNA Polymerase Determine the Stalling at a Deaminated Base

  • Kim, Yun-Jae;Cha, Sun-Shin;Lee, Hyun-Sook;Ryu, Yong-Gu;Bae, Seung-Seob;Cho, Yo-Na;Cho, Hyun-Soo;Kim, Sang-Jin;Kwon, Suk-Tae;Lee, Jung-Hyun;Kang, Sung-Gyun
    • Journal of Microbiology and Biotechnology
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    • v.18 no.8
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    • pp.1377-1385
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    • 2008
  • The uracil-sensing domain in archaeal family B-type DNA polymerases recognizes pro-mutagenic uracils in the DNA template, leading to stalling of DNA polymerases. Here, we describe our new findings regarding the molecular, mechanism underpinning the stalling of polymerases. We observed that two successive deaminated bases were required to stall TNA1 and KOD1 DNA polymerases, whereas a single deaminated base was enough for stalling Pfu DNA polymerase, in spite of the virtually identical uracil-sensing domains. TNA1 and KOD1 DNA polymerases have a much higher extension rate than Pfu DNA polymerase; decreasing the extension rate resulted in stalling by TNA1 and KOD1 DNA polymerases at a single deaminated base. These results strongly suggest that these polymerases require two factors to stop DNA polymerization at a single deaminated base: the presence of the uracil-sensing domain and a relatively slow extension rate.

Comparative LCA of three types of Interior Panel (IP) in Electric Motor Unit (EMU) (전동차 내장패널(Interior Panel)에 대한 비교 전과정평가)

  • Choi, Yo-Han;Lee, Sang-Yong;Kim, Yong-Ki;Lee, Kun-Mo
    • Journal of the Korean Society for Railway
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    • v.10 no.5
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    • pp.596-599
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    • 2007
  • A comparative Life Cycle Assessment (LCA) among three types of Electric Motor Unit (EMU) Interior Panel (IP) was conducted. A functional unit for comparative LCA is a weight of IP for 1 EMU. It is assumed that Manufacturing stage and its upstream processes, Use stage and End of Life (EoL) stage are included in the boundary of product system. For Use stage, the weight of IP causes electricity consumption. It is assumed that aluminum IP is recycled and the other IPs are incinerated at the EoL stage. As a comparison results, aluminum IP has much larger environmental impact (5.162pt) than others (FRP IP; 4.069pt, Phenol IP; 4.053pt) even though recycling consideration is included. The manufacturing stage of aluminum IP has relative big environmental impact (1.824pt) and this point make the most important difference from other IPs (FRP IP; 0.1617pt, Phenol IP; 0.4534pt)). Despite of large weight difference between FRP IP (888.96kg) and phenol IP (316kg), the final environmental impact result has only little difference (0.016pt, 0.39%). With this result, the EMU designer can choose IP with a consideration of the environmental performance of IP.