• Title/Summary/Keyword: single-level cell

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Identification of Cell Type-Specific Effects of DNMT3A Mutations on Relapse in Acute Myeloid Leukemia

  • Seo-Gyeong Bae;Hyeoung-Joon Kim;Mi Yeon Kim;Dennis Dong Hwan Kim;So-I Shin;Jae-Sook Ahn;Jihwan Park
    • Molecules and Cells
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    • v.46 no.10
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    • pp.611-626
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    • 2023
  • Acute myeloid leukemia (AML) is a heterogeneous disease caused by distinctive mutations in individual patients; therefore, each patient may display different cell-type compositions. Although most patients with AML achieve complete remission (CR) through intensive chemotherapy, the likelihood of relapse remains high. Several studies have attempted to characterize the genetic and cellular heterogeneity of AML; however, our understanding of the cellular heterogeneity of AML remains limited. In this study, we performed single-cell RNA sequencing (scRNAseq) of bone marrow-derived mononuclear cells obtained from same patients at different AML stages (diagnosis, CR, and relapse). We found that hematopoietic stem cells (HSCs) at diagnosis were abnormal compared to normal HSCs. By improving the detection of the DNMT3A R882 mutation with targeted scRNAseq, we identified that DNMT3A-mutant cells that mainly remained were granulocyte-monocyte progenitors (GMPs) or lymphoid-primed multipotential progenitors (LMPPs) from CR to relapse and that DNMT3A-mutant cells have gene signatures related to AML and leukemic cells. Copy number variation analysis at the single-cell level indicated that the cell type that possesses DNMT3A mutations is an important factor in AML relapse and that GMP and LMPP cells can affect relapse in patients with AML. This study advances our understanding of the role of DNMT3A in AML relapse and our approach can be applied to predict treatment outcomes.

Evaluation of DNA damage in Pesticide Sprayers using Single Cell Gel Electrophoresis (단세포전기영동법(single Cell Gel Electrophoresis Assay)을 이용한 농약 살포자의 DNA손상 평가)

  • 이연경;이도영;이은일;이동배;류재천;김해준;설동근
    • Environmental Mutagens and Carcinogens
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    • v.21 no.2
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    • pp.128-134
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    • 2001
  • Single cell gel electrophoresis (SCGE) assay, also called comet assay, is a rapid and sensitive method to detect DNA damage in single cell level. To evaluate the DNA damage of lymphocytes of pesticides sprayers, SCGE assay was carried out for 50 pesticides sprayer and 58 control subjects. They were interviewed with structured questionnaire to get the information about the kinds and amount of pesticide. Insecticides and fungicides were predominant among pesticides. Major components of pesticides were organophosphorus, organosulfate, cartap, carbamates, and triazole. Sprayed pesticides were classified into two groups. Group I included organophosphorus, organoarsenic, organotin, tetrazine, triazole and gramoxone, which were known to cause DNA damages. Group II pesticide were carbamates, surfactants, organosulfates, etc., which were not found as DNA damaging agents in scientific documents. Olive tail moments of 100 lymphocytes were measured by KOMET 3.1 program for each person. The means of tail moments were compared between farmers exposed to pesticides and control subjects. Farmers showed higher tail moments than control subjects (2.07$\pm$1.40 vs 1.53$\pm$0.77, p<0.05). The means of tail moments also were compared among group I sprayers (n=36), group II sprayers (n=24) and, control subject, and the means or tail moments were 3.4s$\pm$3.2o, 2.66$\pm$2.20 and 1.53$\pm$0.77 respectively. The difference between means of group I sprayers and controls was statistically significant (p<0.05). In conclusion, this study showed higher DNA damage in farmers exposed to pesticides than control subjects, and comet assay could be useful as a biological monitoring method of genotoxic pesticides for farmers.

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Chemopreventive Effect of Vegetable or Fruit Extract Against Total Diesel Exhaust Particle Extract in NIH/3T3 Cells Using Alkaline Single Cell Gel Electrophoresis (총 디젤분진의 DNA 손상작용과 야채 및 과일추출물의 보호효과)

  • Heo Chan;Kim Nam-Yee;Heo Moon-Young
    • Environmental Analysis Health and Toxicology
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    • v.21 no.2 s.53
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    • pp.127-138
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    • 2006
  • In urban areas, diesel exhaust particles (DEP) are probably a major component of particulate matters, especially in Korea where drive many diesel vehicles. The aim of this study was to investigate genotoxic effects of DEP using single ceil gel electrophoresis. In order to evaluate the mechanisms of DEP genotoxicity, the rat microsome mediated and DNA repair enzyme treated comet assays together with conventional comet assay were performed. Total diesel particles (DEPT) was collected without site fractionation from diesel engine bus and dichloromethane extract was obtained. The organic extract of DEPT revealed DNA damage itself in NIH/3T3 cells. The level of DNA breaks plus oxidative DNA lesions and microsome mediated DNA damage was assessed by modified single cell gel eletrophoresis. DEPT was able to induce oxidative DNA damage as well as microsome mediated DNA damage. Vitamin C as an model antioxidant reduced DNA damage in endonuclase III treated comet assay. One of flavonoid, galangin as a CYP1A1 inhibitor. reduced DNA damage in the presence of S-9 mixture. $DEP_T$ is the sources of oxidative stress, but antioxidants can significantly reduce oxidative DNA dmage. And $DEP_T$ may contain indirect mutagens which can be inhibited by CYP1A1 inhibitors. The ethanol extracts of the mixed vegetables (BV) or the mixed fruits (BF) were evaluated for their in vitro antigenotoxic effects. BV and BF showed potent Inhibitory effects against DEPT induced DNA damage with oxidative DNA lesions and in the prescence of S-9 mixture. These results indicate that BV and BF could prevent cellular DNA damage by inhibiting oxidative stress and suppressing cytochrome P4501A1 in cell culture.

Protoplast Production from Sphacelaria fusca (Sphacelariales, Phaeophyceae) Using Commercial Enzymes

  • Avila-Peltroche, Jose;Won, Boo Yeon
    • Journal of Marine Bioscience and Biotechnology
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    • v.12 no.1
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    • pp.50-58
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    • 2020
  • Sphacelaria is a filamentous brown algal genus that can be epibiotic on macroalgae, marine plants, and sea turtles. Its important role in benthic ecosystems, exposure to different stressors (e.g., grazing), and use as a model organism make Sphacelaria ideal for assessing physiological responses of organisms to environmental inputs. Single-cell RNA sequencing is a powerful new probe for understanding environmental responses of organisms at the molecular (transcriptome) level, capable of delineating gene regulation in different cell types. In the case of plants, this technique requires protoplasts ("naked" plant cells). The existing protoplast isolation protocols for Sphacelaria use non-commercial enzymes and are low-yielding. This study is the first to report the production of protoplasts from Sphacelaria fusca (Hudson) S.F. Gray, using a combination of commercial enzymes, chelation, and osmolarity treatment. A simple combination of commercial enzymes (cellulase Onozuka RS, alginate lyase, and driselase) with chelation pretreatment and an increased osmolarity (2512 mOsm/L H2O) gave a protoplast yield of 15.08 ± 5.31 × 104 protoplasts/g fresh weight, with all the Sphacelaria cell types represented. Driselase had no crucial effect on the protoplast isolation. However, the increased osmolarity had a highly significant and positive effect on the protoplast isolation, and chelation pretreatment was essential for optimal protoplast yield. The protocol represents a significant step forward for studies on Sphacelaria by efficiently generating protoplasts suitable for cellular studies, including single-cell RNA sequencing and expression profiling.

Analysis of Radiation Dose on Single Cells Using Therapeutic Radioisotopes Using the Monte Carlo Method (몬테카를로 방법을 이용한 치료용 방사성동위원소 사용 시 단일 세포에 대한 선량 분석)

  • Kim, Jung-Hoon;Kim, Yu-Soo
    • Journal of radiological science and technology
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    • v.45 no.5
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    • pp.433-438
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    • 2022
  • Targeted radionuclides treatment (TRT) requires the establishment of treatment plans that consider various factors, such as the type of radionuclides, target organs, and administration methods. For this reason, in this study, the absorption dose of a single cell was analyzed according to the type of radioisotope used to treat target radionuclides. In this study, a simulation was performed on beta rays used in the treatment of target radionuclides at the cell level using MCNPX (ver. 2.5.0). First, according to the calculation formula, the beam path according to the type of radioisotope for treatment was calculated. Second, the amount of self-radiation by beta rays emitted from cell diameters of 5 ㎛ and 10 ㎛ cell nuclei was evaluated. As a result, it showed a high range proportional to the maximum energy of the beta-ray, and the highest self-dose distribution from 177 Lu radiation sources among therapeutic radioisotopes. This was analyzed as a result that is inversely proportional to the maximum energy of the beta-ray, and it suggests that the selection of a nuclide considering the range of the beta-ray is necessary in the treatment of target radionuclides in the future.

MLC NAND-type Flash Memory Built-In Self Test for research (MLC NAND-형 Flash Memory 내장 자체 테스트에 대한 연구)

  • Kim, Jin-Wan;Kim, Tae-Hwan;Chang, Hoon
    • Journal of the Institute of Electronics and Information Engineers
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    • v.51 no.3
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    • pp.61-71
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    • 2014
  • As the occupancy rate of the flash memory increases in the storage media market for the embedded system and the semi-conductor industry grows, the demand and supply of flash memory is increasing by a big margin. They are especially used in large quantity in the smart phones, tablets, PC, SSD and Soc(System on Chip) etc. The flash memory is divided into the NOR type and NAND type according to the cell arrangement structure and the NAND type is divided into the SLC(Single Level Cell) and MLC(Multi Level Cell) according to the number of bits that can be stored in each cell. Many tests have been performed on NOR type such as BIST(Bulit-In Self Test) and BIRA(Bulit-In Redundancy Analysis) etc, but there is little study on the NAND type. For the case of the existing BIST, the test can be proceeded using external equipments like ATE of high price. However, this paper is an attempt for the improvement of credibility and harvest rate of the system by proposing the BIST for the MLC NAND type flash memory of Finite State Machine structure on which the pattern test can be performed without external equipment since the necessary patterns are embedded in the interior and which uses the MLC NAND March(x) algorithm and pattern which had been proposed for the MLC NAND type flash memory.

Effects of Monosodium Glutamate on Unscheduled DNA Synthesis and DNA Single-Strand Breaks in Primary Cultures of Rat Hepatocytes (일차배양 간세포에서 Monosodium Glutamate에 의한 돌연변이 유발성의 검증)

  • 김동현;양규환
    • Environmental Mutagens and Carcinogens
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    • v.7 no.2
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    • pp.59-64
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    • 1987
  • Cytotoxic and genotoxic potential of monosodium glutamate (MSG) were evaluated in primary cultures of rat hepatocytes. When exposed to liver cell culture continuously for 24 hr, MSG did not show any cytotoxic effects up to 0.5% (w/v) level as determined by Tryphan Blue exclusion and lactic dehydrogenase release test. MSG also did not induce unscheduled DNA synthesis or DNA single-strand breaks in hepatocyte cultures up to 1% level. No synergistic effects of MSG were observed on aflatoxin B$_1$-induced DNA damage when 1% MSG was treated to liver cell culture along with aflatoxin B$_1$.

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An Efficient SLC Transition Method for Improving Defect Rate and Longer Lifetime on Flash Memory (플래시 메모리 상에서 불량률 개선 및 수명 연장을 위한 효율적인 단일 비트 셀 전환 기법)

  • Hyun-Seob Lee
    • Journal of Internet of Things and Convergence
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    • v.9 no.3
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    • pp.81-86
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    • 2023
  • SSD (solid state disk), which is flash memory-based storage device, has the advantages of high density and fast data processing. Therefore, it is being utilized as a storage device for high-capacity data storage systems that manage rapidly increasing big data. However, flash memory, a storage media, has a physical limitation that when the write/erase operation is repeated more than a certain number of times, the cells are worn out and can no longer be used. In this paper, we propose a method for converting defective multi-bit cells into single-bit cells to reduce the defect rate of flash memory and extend its lifetime. The proposed idea distinguishes the defects and treatment methods of multi-bit cells and single-bit cells, which have different physical characteristics but are treated as the same defect, and converts the expected defective multi-bit cells into single-bit cells to improve the defect rate and extend the overall lifetime. Finally, we demonstrate the effectiveness of our proposed idea by measuring the increased lifetime of SSD through simulations.

Multilevel multiuser detection system in multi-cell MFSK/FH-CDMA environment

  • ;Ryuji Kohno
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.23 no.4
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    • pp.865-872
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    • 1998
  • This paper proposes a multiuse detection system in a multi-cell M-ary Frequency Shift Keying(MFSK)/frequency hopping(FH)-Code Division Multiple Access(CDMA) environment, in which the channel model is an OR-channel and in the reverse link. We have proposed a multiuse detection system in a single cell. However, this sitye is not adequate to detect multiuser in a multi-cell environment. Therefore, we propose a multiuser detection system based on 3 level OR decision with two threholds. The proposed detection system can delete interference as well as intra-cell interference, receive the weakened desired signal and reject the false alarm. computer simulation shows the performance improvement.

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Differential Effects of Nongenotoxic and Genotoxic Carcinogen on Cell Proliferation and c-Jun Expression in the Rat Liver Initiated with Diethylnitrosamine

  • Kim, Hye-Jin;Kim, Jong-Won;Hong, Jin-Tae;Nam, Ki-Taek;Kim, Dae-Joong
    • Environmental Mutagens and Carcinogens
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    • v.19 no.2
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    • pp.89-94
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    • 1999
  • Cell proliferation and c-Jun expression pattern in liver exposed by nongenotoxic carcinogens phenobarbital (PB) and clofibrate, and genotoxic carcinogen 2-amino-3-methylimidazo [4,5-f] quinoline (IQ) were investigated to see whether differential effects of genotoxic and non-genotoxic carcinogens on the development of neoplastic foci may be related to differential effect on cell proliferation. Male F344 rats were initially given a single intraperitioneal injection of diethylnitrosamine (200 mg/kg body weight), and 2 weeks later, animals were fed diets containing 0.03% IQ or 0.5% CE or 0.05% PB or basal diet as a control for 6 weeks. All rats were subjected to the two-thirds partial hepatectomy (PH) at week 3. Sequential sacrifice of rats was performed until 8 weeks. Cell proliferation was examined by immunohistochemical staining of bromodeoxyuridine and c-Jun expression was determined by northern blotting. The increase of cell proliferation rate after PH was significant in the rats fed 0.05% IQ and continued until 8 weeks, while the increase was not significant in the rats fed phenobarbital and clofibrate compared to that in the rats fed control diet. mRNA level of c-Jun in the liver treated with IQ was about 7 fold higher than that of control and peak at 5 hours after rH. In the liver treated with CE, mRNA level of c-Jun was 3-4 fold higher than that of control and the highest level of mRNA of c-Jun was seen at 24 hours after PH. These results show that differential effects of genotoxic and non-genotoxic carcinogens on the development of neoplastic foci may be related to differential effect on cell proliferation pattern.

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