• Title/Summary/Keyword: high-throughput biology

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Proteome in Toxicological Assessment of Endocrine Disrupting Chemicals (프로테오믹스를 이용한 내분비계 교란물질 환경독성 연구)

  • 김호승;계명찬
    • Korean Journal of Environmental Biology
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    • v.21 no.2
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    • pp.87-100
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    • 2003
  • It is important to understand the potential human health implications of exposure to environmental chemicals that may act as hormonally active agents. It is necessary to have an understanding of how pharmaceutical and personal care products and other chemicals affect the ecosystem of our planet as well as human health. Endocrine disruption is defined as the ability of a chemical contaminating the workplace or the environment to interfere with homeostasis, development, reproduction, and/or behavior in a living organism or it's offspring. Certain classes of environmentally persistent chemicals such as polychlorinated biphenyls (PCBs), dioxins, furans, and some pesticides can adversely effect the endocrine systems of aquatic life and terrestrial wildlife. Research continues to support the theory of endocrine disruption. However, endocrine disruption researches have been applied to proteomics poorly. Proteomics can be defined as the systematic analysis of proteins for their identity, quantity and function. It could increase the predictability of early drug development and identify non-invasive biomarkers of tonicity or efficacy. Proteome analysis is most commonly accomplished by the combination of two-dimensional gel electrophoresis (2D/E) and MALDI-TOF mass spectrometry (MS) sr protein chip array and SELDI-TOF MS. Proteomics have an opportunity to play an important role in resolving the question of what role endocrine disruptors play in initiating human disease. Proteomics can also play an imfortant role in the evaluation of the risk assessment and use of risk management and risk communication tools required to address public health concerns related to notions of endocrine disruptors. Understanding the need for the proteomics and possessing knowledge of the developing biomakers used to abbess endocrine activity potential will he essential components relevant to the topic of endocrine disruptors.

The Effects of Cesium, Strontium and Cobalt on Cell Toxicity in the 2D and 3D Cell Culture Platforms (단층 및 입체 세포배양환경에서 세슘, 스트론튬 및 코발트가 세포 독성에 미치는 영향 분석)

  • Kim, Gi Yong;Kang, Sung-Min;Jang, Sung-Chan;Huh, Yun Suk;Roh, Changhyun
    • Korean Journal of Environmental Biology
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    • v.34 no.2
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    • pp.107-115
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    • 2016
  • Currently, there are 442 operating nuclear power plants in the world, and 62 more are under construction. According to this reasoning, the treatment of radioactive waste is important to prevent the environmental ecosystem including humans, animals, and plants. Especially, a leakage of radioactive waste causes not only regional problem but also serious global one. In this study, we demonstrate the effect of radioisotopes (e.g., cesium, strontium, and cobalt) on a 3D culture cell. To develop the 3D cell culture system, we used a 96-well-culture plate with biocompatible agarose hydrogel. Using this method, we can perform the 3D cell culture system with three different cell lines such as HeLa, HepG2, and COS-7. In addition, we conducted a cell viability test in the presence of radioisotopes. Interestingly, the 3D morphological cells showed 42% higher cell viability than those on the 2D against cesium. This result indicates that the 3D platform provides cells morphological and physiological characteristic similar to in vivo grown tissues. Moreover, it overcomes the limitation of conventional cell culture system that can't reflect in vivo systems. Finally, we believe that the proposed approach can be applied a new strategy for simple high-throughput screening and accurate evaluation of metal toxicity assay.

Establishment of the large-scale longitudinal multi-omics dataset in COVID-19 patients: data profile and biospecimen

  • Jo, Hye-Yeong;Kim, Sang Cheol;Ahn, Do-hwan;Lee, Siyoung;Chang, Se-Hyun;Jung, So-Young;Kim, Young-Jin;Kim, Eugene;Kim, Jung-Eun;Kim, Yeon-Sook;Park, Woong-Yang;Cho, Nam-Hyuk;Park, Donghyun;Lee, Ju-Hee;Park, Hyun-Young
    • BMB Reports
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    • v.55 no.9
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    • pp.465-471
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    • 2022
  • Understanding and monitoring virus-mediated infections has gained importance since the global outbreak of the coronavirus disease 2019 (COVID-19) pandemic. Studies of high-throughput omics-based immune profiling of COVID-19 patients can help manage the current pandemic and future virus-mediated pandemics. Although COVID-19 is being studied since past 2 years, detailed mechanisms of the initial induction of dynamic immune responses or the molecular mechanisms that characterize disease progression remains unclear. This study involved comprehensively collected biospecimens and longitudinal multi-omics data of 300 COVID-19 patients and 120 healthy controls, including whole genome sequencing (WGS), single-cell RNA sequencing combined with T cell receptor (TCR) and B cell receptor (BCR) sequencing (scRNA(+scTCR/BCR)-seq), bulk BCR and TCR sequencing (bulk TCR/BCR-seq), and cytokine profiling. Clinical data were also collected from hospitalized COVID-19 patients, and HLA typing, laboratory characteristics, and COVID-19 viral genome sequencing were performed during the initial diagnosis. The entire set of biospecimens and multi-omics data generated in this project can be accessed by researchers from the National Biobank of Korea with prior approval. This distribution of large-scale multi-omics data of COVID-19 patients can facilitate the understanding of biological crosstalk involved in COVID-19 infection and contribute to the development of potential methodologies for its diagnosis and treatment.

Establishment of a NanoBiT-Based Cytosolic Ca2+ Sensor by Optimizing Calmodulin-Binding Motif and Protein Expression Levels

  • Nguyen, Lan Phuong;Nguyen, Huong Thi;Yong, Hyo Jeong;Reyes-Alcaraz, Arfaxad;Lee, Yoo-Na;Park, Hee-Kyung;Na, Yun Hee;Lee, Cheol Soon;Ham, Byung-Joo;Seong, Jae Young;Hwang, Jong-Ik
    • Molecules and Cells
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    • v.43 no.11
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    • pp.909-920
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    • 2020
  • Cytosolic Ca2+ levels ([Ca2+]c) change dynamically in response to inducers, repressors, and physiological conditions, and aberrant [Ca2+]c concentration regulation is associated with cancer, heart failure, and diabetes. Therefore, [Ca2+]c is considered as a good indicator of physiological and pathological cellular responses, and is a crucial biomarker for drug discovery. A genetically encoded calcium indicator (GECI) was recently developed to measure [Ca2+]c in single cells and animal models. GECI have some advantages over chemically synthesized indicators, although they also have some drawbacks such as poor signal-to-noise ratio (SNR), low positive signal, delayed response, artifactual responses due to protein overexpression, and expensive detection equipment. Here, we developed an indicator based on interactions between Ca2+-loaded calmodulin and target proteins, and generated an innovative GECI sensor using split nano-luciferase (Nluc) fragments to detect changes in [Ca2+]c. Stimulation-dependent luciferase activities were optimized by combining large and small subunits of Nluc binary technology (NanoBiT, LgBiT:SmBiT) fusion proteins and regulating the receptor expression levels. We constructed the binary [Ca2+]c sensors using a multicistronic expression system in a single vector linked via the internal ribosome entry site (IRES), and examined the detection efficiencies. Promoter optimization studies indicated that promoter-dependent protein expression levels were crucial to optimize SNR and sensitivity. This novel [Ca2+]c assay has high SNR and sensitivity, is easy to use, suitable for high-throughput assays, and may be useful to detect [Ca2+]c in single cells and animal models.

Effect of Culture Media on Production of Biomass, Fatty Acid, and Carotenoid in a Newly Isolated Mychonastes sp. (신규 분리된 Mychonastes sp.의 생장, 지방산 및 색소 생산에 생장배지가 미치는 영향)

  • Yim, Kyung June;Jang, Hyun-Jin;Park, Yeji;Nam, Seung Won;Hwang, Byung Su;Jung, Ji Young;Lee, Chang Soo;Kim, Z-Hun
    • Journal of Marine Bioscience and Biotechnology
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    • v.14 no.1
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    • pp.1-8
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    • 2022
  • This study examined the growth, fatty acid (FA) content, and carotenoids of a newly isolated freshwater microalga, Mychonastes sp. 246, in various culture media. The appropriate temperature and light intensity for culturing Mychonastes sp. 246 were determined as 18℃-22℃ and 200-250 µmol/m2/s using a high throughput photobioreactor. The microalgal cells were cultivated in 0.5 L bubble column photobioreactors using BG11, Bold's Basal media, and f/2 media. According to the growth results of the microalgae, BG11, among the tested media, showed the highest biomass concentrations (3.5 ± 0.1 g/L in 10 d). To enhance the biomass growth of the microalgae, the N:P ratio in BG11 was manipulated from 45:1 to 7:1 based on the stoichiometric cell composition. The biomass concentrations of Mychonastes sp. 246 grown on the manipulated BG11 (MBG) increased to 38% (4.6 ± 0.3 g/L in d) compared with the original BG11 (3.3 g/L). The FA content of the microalgae grown on the MBG was lower (8.4%) than that of the original BG11 (10.1%) while the FA compositions did not exhibit any significant differences. Furthermore, three kinds of carotenoids were identified in Mychonastes sp. 246, zeaxanthin, lutein, and β-carotene. These results suggest an effective strategy for increasing biomass concentrations, FA content, and carotenoids of microalgae by performing a simple N:P adjustment in the culture media.