• Title/Summary/Keyword: Cell analysis

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LAMMER Kinase Modulates Cell Cycle by Phosphorylating the MBF Repressor, Yox1, in Schizosaccharomyces pombe

  • Kibum Park;Joo-Yeon Lim;Je-Hoon Kim;Jieun Lee;Songju Shin;Hee-Moon Park
    • Mycobiology
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    • v.51 no.5
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    • pp.372-378
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    • 2023
  • Lkh1, a LAMMER kinase homolog in the fission yeast Schizosaccharomyces pombe, acts as a negative regulator of filamentous growth and flocculation. It is also involved in the response to oxidative stress. The lkh1-deletion mutant displays slower cell growth, shorter cell size, and abnormal DNA content compared to the wild type. These phenotypes suggest that Lkh1 controls cell size and cell cycle progression. When we performed microarray analysis using the lkh1-deletion mutant, we found that only four of the up-regulated genes in the lkh1-deletion were associated with the cell cycle. Interestingly, all of these genes are regulated by the Mlu1 cell cycle box binding factor (MBF), which is a transcription complex responsible for regulating the expression of cell cycle genes during the G1/S phase. Transcription analyses of the MBF-dependent cell-cycle genes, including negative feedback regulators, confirmed the up-regulation of these genes by the deletion of lkh1. Pull-down assay confirmed the interaction between Lkh1 and Yox1, which is a negative feedback regulator of MBF. This result supports the involvement of LAMMER kinase in cell cycle regulation by modulating MBF activity. In vitro kinase assay and NetPhosK 2.0 analysis with the Yox1T40,41A mutant allele revealed that T40 and T41 residues are the phosphorylation sites mediated by Lkh1. These sites affect the G1/S cell cycle progression of fission yeast by modulating the activity of the MBF complex.

Optimization of Culture Medium for Lactosucrose ($^4G-{\beta}$-D-Galactosylsucrose) Production by Sterigmatomyces elviae Mutant Using Statistical Analysis

  • Lee, Jong-Ho;Lim, Jung-Soo;Song, Yoon-Seok;Kang, Seong-Woo;Prak, Chul-Hwan;Kim, Seung-Wook
    • Journal of Microbiology and Biotechnology
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    • v.17 no.12
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    • pp.1996-2004
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    • 2007
  • In this study, the optimization of culture medium using a Sterigmatomyces elviae mutant was investigated using statistical analysis to increase the cell mass and lactosucrose ($^4G-{\beta}$-D-galactosylsucrose) production. In basal medium, the cell mass and lactosucrose production were 4.12 g/l and 140.91 g/l, respectively. However, because of the low cell mass and lactosucrose production, optimization of culture medium was carried out to increase the cell mass and lactosucrose production. Culture media were optimized by the S. elviae mutant using analysis of variance (ANOVA) and response surface methodology (RSM). Central composite designs using RSM were utilized in this investigation. Quadratic models were obtained for cell mass and lactosucrose production. In the case of cell mass, optimal components of the medium were as follows: sucrose 1.13%, yeast extract 0.99%, bactopeptone 2.96%, and ammonium sulfate 0.40%. The predicted maximum value of cell mass was about 5.20 g/l and its experimental value was 5.08 g/l. In the case of lactosucrose production, optimal components of the medium were as follows: sucrose 0.96%, yeast extract 1.2%, bactopeptone 3.0%, and ammonium sulfate 0.48%. Then, the predicted maximum value of lactosucrose production was about 194.12 g/l and the corresponding experimental value was about 183.78 g/l. Therefore, by culturing using predicted conditions, the real cell mass and lactosucrose production increased to 23.3% and 30.42%, respectively.

Identification of Sugar-Responsive Genes and Discovery of the New Functions in Plant Cell Wall

  • Lee, Eun-Jeong
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2007.04a
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    • pp.65-73
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    • 2007
  • The objective of this study is to understand how regulatory mechanisms respond to sugar status for more efficient carbon utilization and source-sink regulation in plants. So, we need to identify and characterize many components of sugar-response pathways for a better understanding of sugar responses. For this end, genes responding change of sugar status were screened using Arabidpsis cDNA arrays, and confirmed thirty-six genes to be regulated by sucrose supply in detached leaves by RNA blot analysis. Eleven of them encoding proteins for amino acid metabolism and carbohydrate metabolism were repressed by sugars. The remaining genes induced by sugar supply were for protein synthesis including ribosomal proteins and elongation factors. Among them, I focused on three hydrolase genes encoding putative $\beta$-galactosidase, $\beta$-xylosidase, and $\beta$-glucosidase that were transcriptionally induced in sugar starvation. Homology search indicated that these enzymes were involved in hydrolysis of cell wall polysaccharides. In addition to my results, recent transcriptome analysis suggested multiple genes for cell wall degradation were induced by sugar starvation. Thus, I hypothesized that enzyme for cell wall degradation were synthesized and secreted to hydrolyze cell wall polysaccharides producing carbon source under sugar-starved conditions. In fact, the enzymatic activities of these three enzymes increased in culture medium of Arabidopsis suspension cells under sugar starvation. The $\beta$-galactosidase encoded by At5g56870 was identified as a secretory protein in culture medium of suspension cells by mass spectrometry analysis. This protein was specifically detected under sugar-starved condition with a specific antibody. Induction of these genes was repressed in suspension cells grown with galactose, xylose and glucose as well as with sucrose. In planta, expression of the genes and protein accumulation were detected when photosynthesis was inhibited. Glycosyl hydrolase activity against galactan also increased during sugar starvation. Further, contents of cell wall polysaccharides especially pectin and hemicellulose were markedly decreased associating with sugar starvation in detached leaves. The amount of monosaccharide in pectin and hemicellulose in detached leaves decreased in response to sugar starvation. These results supported my idea that cell wall has one of function to supply carbon source in addition to determination of cell shape and physical support of plant bodies.

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in vitro Assessment of Antineoplastic Effects of Deuterium Depleted Water

  • Soleyman-Jahi, Saeed;Zendehdel, Kazem;Akbarzadeh, Kambiz;Haddadi, Mahnaz;Amanpour, Saeid;Muhammadnejad, Samad
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.5
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    • pp.2179-2183
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    • 2014
  • Background: In vitro, in vivo and clinical studies have demonstrated anti-cancer effects of deuterium depleted water (DDW). The nature of this agents action, cytotoxic or cytostatic, remains to be elucidated. We here aimed to address the point by examining effects on different cell lines. Materials and Methods: 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) -based cytotoxicity analysis was conducted for human breast, stomach, colon, prostate cancer and glioblastoma multiforme cell lines as well as human dermal fibroblasts. The cell lines were treated with decreasing deuterium concentrations of DDW alone, paclitaxel alone and both. One way analysis of variance (ANOVA) was used for statistical analysis. Results: Treatment with different deuterium concentrations of DDW alone did not impose any significant inhibitory effects on growth of cell lines. Paclitaxel significantly decreased the survival fractions of all cell lines. DDW augmented paclitaxel inhibitory effects on breast, prostate, stomach cancer and glioblastoma cell lines, with influence being more pronounced in breast and prostate cases. Conclusions: DDW per se does not appear to have inhibitory effects on the assessed tumor cell lines as well as normal fibroblasts. As an adjuvant, however, DDW augmented inhibitory effects of paclitaxel and thus it could be considered as an adjuvant to conventional anticancer agents in future trials.

The effect of β-sitosterol proliferation and apoptosis in human uterine leiomyoma cells (계혈등(鷄血藤)의 Beta-sitosterol 성분이 자궁근종세포의 증식억제와 세포자멸사의 유도에 미치는 영향)

  • Park, Youngsun;Baek, Seunghee
    • The Journal of Korean Obstetrics and Gynecology
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    • v.18 no.1
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    • pp.181-191
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    • 2005
  • Purpose : ${\beta}$-sitosterol is kind of phytosterols or plant which are structurally similar to cholesterol. This study was aimed to investigate the inhibitory effect of the ${\beta}$-sitosterol on the proliferation of human uterine leiomyoma cells and the expression of gene related the mechanism of cell apoptosis. Methods : We counted the number of death cells treated with indicated time of the ${\beta}$-sitosterol and investigated cell death rate by cell count assay. Furthermore, flow cytometry analysis and DNA fragmentation assay were used to dissect between necrosis and apoptosis. and then we observed the differential gene expression by western blot analysis. Results : 1) The inhibitory effect on the growth of uterine leiomyoma cell treated with the ${\beta}$-sitosterol $16{\mu}M$ was increased in a time dependent. 2) The result of flow cytometry analysis, subG1 phase arrest related cell apoptosis was investigated 16.97% in uterine leiomyoma cell treated with the ${\beta}$-sitosterol $16{\mu}M$ and showed the fashion of proportional time dependent. 3) The gene expression of p27, p21 related cell cycle was increased according to increasing time interval but cyclin E-CDK2 complex was decreased expression. 4) The character of apoptosis, DNA fragmentation was significantly observed on the time dependent. 5) The expression of pro-caspase 3 and PARP were decreased dependent on treatment with time dependent. Conclusion : This study showed that the ${\beta}$-sitosterol have the inhibitory effect on the proliferation of human uterine leiomyoma cell and the effect was related with apoptosis.

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Analysis of the Deformed Unit Cell by Clamping Force Through the FEM and CFD Interaction (FEM과 CFD 연동을 통한 스택 체결 시 압력에 의해 변형된 단위 전지 해석)

  • YOO, BIN;LIM, KISUNG;JU, HYUNCHUL
    • Transactions of the Korean hydrogen and new energy society
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    • v.32 no.4
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    • pp.228-235
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    • 2021
  • Polymer electrolyte membrane fuel cells (PEMFC) are currently being used in various transport applications such as drones, unmanned aerial vehicles, and automobiles. The power required is different according to the type of use, purpose, and the conditions adjusted using a cell stack. The fuel cell stack is compressed to reduce the size and prevent fuel leakage. The unit cells that make up the cell stack are subjected to compression by clamping force, which makes geometrical changes in the porous media and it impacts on cell performance. In this study, finite elements method (FEM) and computational fluid dynamics (CFD) analysis for the deformed unit cell considering the effects of clamping force is performed. First, structural analysis using the FEM technique over the deformed gas diffusion layer (GDL) considering compression is carried out, and the resulting porosity changed in the GDL is calculated. The PEMFC model is then verified by a three-dimensional, two-phase fuel cell simulation applying the physical properties and geometry obtained before and after compression. The detailed simulation results showed different concentration distributions of fuel between the original and deformed geometry, resulting in the difference in the distribution of current density is represented at compressed GDL region with low oxygen concentration.

Engineered microdevices for single cell immunological assay

  • Choi, Jong-Hoon
    • Interdisciplinary Bio Central
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    • v.2 no.2
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    • pp.1.1-1.8
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    • 2010
  • Microdevices have been used as effective experimental tools for the rapid and multiplexed analysis of individual cells in single-cell assays. Technological advances for miniaturizing such systems and the optimization of delicate controls in micron-sized space homing cells have motivated many researchers from diverse fields (e.g., cancer research, stem cell research, therapeutic agent development, etc.) to employ microtools in their scientific research. Microtools allow high-throughput, multiplexed analysis of single cells, and they are not limited by the lack of large samples. These characteristics may significantly benefit the study of immune cells, where the number of cells available for testing is usually limited. In this review, I present an overview of several microtools that are currently available for single-cell analyses in two popular formats: microarrays and microfluidic microdevices. Then, I discuss the potential to study human immunology on the single-cell level, and I highlight several recent examples of immunoassays performed with single-cell microdevice assays. Finally, I discuss the outlook for the development of optimized assay platforms to study human immune cells. The development and application of microdevices for studies on single immune cells presents novel opportunities for the qualitative and quantitative characterization of immune cells and may lead to a comprehensive understanding of fundamental aspects of human immunology.

An Experimental Analysis of the Ripple Current Applied Variable Frequency Characteristic in a Polymer Electrolyte Membrane Fuel Cell

  • Kim, Jong-Hoon;Jang, Min-Ho;Choe, Jun-Seok;Kim, Do-Young;Tak, Yong-Sug;Cho, Bo-Hyung
    • Journal of Power Electronics
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    • v.11 no.1
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    • pp.82-89
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    • 2011
  • Differences in the frequency characteristic applied to a ripple current may shorten fuel cell life span and worsen the fuel efficiency. Therefore, this paper presents an experimental analysis of the ripple current applied variable frequency characteristic in a polymer electrolyte membrane fuel cell (PEMFC). This paper provides the first attempt to examine the impact of ripple current through immediate measurements on a single cell test. After cycling for hours at three frequencies, each polarization and impedance curve is obtained and compared with those of a fuel cell. Through experimental results, it can be absolutely concluded that low frequency ripple current leads to long-term degradation of a fuel cell. Three different PEMFC failures such as membrane dehydration, flooding and carbon monoxide (CO) poisoning that lead to an increase in the impedance magnitude at low frequencies are simply introduced.

Neuroprotective Effects of Some Plant Extracts Against Dopamine-induced Oxidative Stress on Neuronal Cell (Dopamine에 의해 산화적 스트레스를 받은 Neuronal Cell에 뇌 보호 효과를 가지는 수종 생약추출물의 검색)

  • Koo, Uk;Lee, Hak-Ju;Lee, Dong-Ho;Lee, Hyun-Jung;Ham, Ah-Rom;Mar, Woong-Chon
    • Korean Journal of Pharmacognosy
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    • v.40 no.1
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    • pp.41-45
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    • 2009
  • Parkinson's disease (PD) is the second most common neurodegenerative disorder after Alzhemier's disease. Neuropathologically, PD is characterized by the selective loss of dopaminergic neurons. The neuronal toxicity of cytosolic excess dopamine (DA) has been described in many studies using several cell lines. In dopaminergic neurons, cytosolic excess DA is easily oxidized via monoamine oxidase (MAO)-B, tyrosinase or by auto-oxidation to produce neurotoxic metabolites such as DA quinone. So, in the present study, we induced cell death by treatment of DA ($600{\mu}M$) in human neuroblastoma SH-SY5Y cell which was treated samples before 24 hr, and cell viability was measured by fluorescence activated cell sorter (FACs) analysis. Of those tested, the extracts of Poria cocos (赤茯笭)(whole), Gastrodia elata (rhizomes), Eucommia ulmoides (炒)(barks), Syneilesis palmata (whole), Acorus gramineus (rhizomes), Ligustrum japonicum (leaves) showed neuroprotective effects in dose dependent manner.

CFD-based Flow Simulation Study of Fuel Cell Protective Gas (CFD를 활용한 연료전지 모듈 보호가스 유동 연구)

  • Kwon, Kiwook;Lim, Jongkoo;Park, Jongcheol;Shin, Hyun Khil
    • 한국신재생에너지학회:학술대회논문집
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    • 2011.11a
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    • pp.86.1-86.1
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    • 2011
  • To improve the safety, the fuel cell operate inside a pressurized enclosure which contains inert gas so called protective gas. The protective gas not only prevents the mixture of hydrogen and oxygen, but also removes the water in the vessel with the condenser. This study presents the details of the flow optimization in order to reduce the humidity in the fuel cell housing. The protective gas flow in the fuel cell container is studied by Computational Fluid Dynamics(CFD) simulations. This study focuses on optimizing the geometry of an protective gas circulation system in fuel cell module to reduce the humidity in the vessel. CFD analysis was carried out for an existing model to understand the flow behavior through the fuel cell system. Based on existing model CFD results, geometrical changes like inlet placement, optimization of outlet size, modification of fuel cell module system are carried out, to improve the flow characteristics. The CFD analysis of the optimized model is again carried out and the results show good improvement in protective gas flow behavior.

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