• Title/Summary/Keyword: Cell State

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Characterization of Two GAS1 Genes and Their Effects on Expression and Secretion of Heterologous Protein Xylanase B in Kluyveromyces lactis

  • Lian, Zhao;Jiang, Jing-Bo;Chi, Shuang;Guan, Guo-Hua;Li, Ying;Li, Ji-Lun
    • Journal of Microbiology and Biotechnology
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    • v.25 no.12
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    • pp.1977-1988
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    • 2015
  • β-1,3-glucanosyltransferases play essential roles in cell wall biosynthesis in yeast. Kluyveromyces lactis has six putative β-1,3-glucanosyltransferase genes. KlGAS1-1 and KlGAS1-2 are homologs of Saccharomyces cerevisiae gene GAS1. RT-qPCR indicated the transcription level of KlGAS1-1 was significantly reduced while heterologous protein (thermostable xylanase B) secretion was enhanced during medium optimization. To evaluate if these two events were related, and to improve xylanase B secretion in K. lactis, we constructed KlGAS1-1 and KlGAS1-2 single deletion strains and double deletion strain, respectively. KlGAS1-1 gene deletion resulted in the highest xylanase B activity among the three mutants. Only the double deletion strain showed morphology similar to that of the GAS1 deletion mutant in S. cerevisiae. The two single deletion strains differed in terms of cell wall thickness and xylanase B secretion. Transcription levels of β-1,3-glucanosyltransferase genes and genes related to protein secretion and transport were assayed. The β-1,3-glucanosyltransferase genes displayed transcription complementation in the cell wall synthesis process. KlGAS1-1 and KlGAS1-2 affected transcription levels of secretion- and transport-related genes. Differences in protein secretion ratio among the three deletion strains were associated with changes of transcription levels of secretion- and transport-related genes. Our findings indicate that KlGAS1-1 deletion is an effective tool for enhancing industrial-scale heterologous protein secretion in K. lactis.

An Experimental Study on Transient Characteristics of PEM Fuel Cell Stack (PEM 연료전지 스택의 과도상태 출력특성에 관한 실험적 연구)

  • Kim, Hyun-il;Hwang, Jae-Soon;Chung, Tae-Yong;Shin, Dong-Hoon;Nam, Jin-Hyun;Kim, Young-Gyu
    • Proceedings of the KSME Conference
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    • 2007.05b
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    • pp.2003-2008
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    • 2007
  • The transient power characteristics of a PEM fuel cell stack was experimentally studied using a commercial 1.2kW PEM fuel cell ($Nexa^{TM}$ Power Module, Ballard Power System Inc.). The conditions in PEM fuel cell stack such as temperature and water content change rather slowly because of their large heat capacity and long channel length, which results in long transient time to converge to a steady state. The steady characteristics of the PEM fuel cell module was determined first, followed by the measurement of its transient characteristics upon stepwise and continuous load current changes. During the stepwise current change from 5A to 25A, the output voltage initially decreased below the steady voltage and then increased gradually. Similar behavior was also observed for the stepwise current change from 25A to 5A. This transient behavior is explained with reference to the evolution of the temperature and water content of the PEM fuel cell stack.

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miR-15b induced by platelet-derived growth factor signaling is required for vascular smooth muscle cell proliferation

  • Kim, Sunghwan;Kang, Hara
    • BMB Reports
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    • v.46 no.11
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    • pp.550-554
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    • 2013
  • The platelet-derived growth factor (PDGF) signaling pathway is essential for inducing a dedifferentiated state of vascular smooth muscle cells (VSMCs). Activation of PDGF inhibits smooth muscle cell (SMC)-specific gene expression and increases the rate of proliferation and migration, leading to dedifferentiation of VSMCs. Recently, microRNAs have been shown to play a critical role in the modulation of the VSMC phenotype in response to extracellular signals. However, little is known about microRNAs regulated by PDGF in VSMCs. Herein, we identify microRNA- 15b (miR-15b) as a mediator of VSMC phenotype regulation upon PDGF signaling. We demonstrate that miR-15b is induced by PDGF in pulmonary artery smooth muscle cells and is critical for PDGF-mediated repression of SMC-specific genes. In addition, we show that miR-15b promotes cell proliferation. These results indicate that PDGF signaling regulates SMC-specific gene expression and cell proliferation by modulating the expression of miR-15b to induce a dedifferentiated state in the VSMCs.

Hybrid Learning-Based Cell Morphology Profiling Framework for Classifying Cancer Heterogeneity (암의 이질성 분류를 위한 하이브리드 학습 기반 세포 형태 프로파일링 기법)

  • Min, Chanhong;Jeong, Hyuntae;Yang, Sejung;Shin, Jennifer Hyunjong
    • Journal of Biomedical Engineering Research
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    • v.42 no.5
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    • pp.232-240
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    • 2021
  • Heterogeneity in cancer is the major obstacle for precision medicine and has become a critical issue in the field of a cancer diagnosis. Many attempts were made to disentangle the complexity by molecular classification. However, multi-dimensional information from dynamic responses of cancer poses fundamental limitations on biomolecular marker-based conventional approaches. Cell morphology, which reflects the physiological state of the cell, can be used to track the temporal behavior of cancer cells conveniently. Here, we first present a hybrid learning-based platform that extracts cell morphology in a time-dependent manner using a deep convolutional neural network to incorporate multivariate data. Feature selection from more than 200 morphological features is conducted, which filters out less significant variables to enhance interpretation. Our platform then performs unsupervised clustering to unveil dynamic behavior patterns hidden from a high-dimensional dataset. As a result, we visualize morphology state-space by two-dimensional embedding as well as representative morphology clusters and trajectories. This cell morphology profiling strategy by hybrid learning enables simplification of the heterogeneous population of cancer.

The correlation between ionic conductivity and cell performance with various compositions of polymer electrolyte in dye-sensitized solar cells (염료감응형 태양전지에서의 고분자 전해질 종류에 따른 이온전도도와의 상호관계)

  • Cha, Si-Young;Kim, Su-Jin;Lee, Yong-Gun;Kang, Yong-Soo
    • 한국신재생에너지학회:학술대회논문집
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    • 2007.11a
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    • pp.306-308
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    • 2007
  • Poly(ethylene glycol) dimethyl ether (PEGDME)/fumed silica/ 1-methyl -3-propylimidazolium iodide (MPII)/$I_2$ mixtures were used as polymer electrolytes in solid state dye-sensitized solar cells (DSSCs). The contents of MPII were changed and the concentration of $I_2$ was fixed at 0.1 mole% with respect to the MPII. The maximum ionic conductivity was obtained at [EG]:[MPII]:[$I_2$]=10:1.5:0.15. It was supposed that the maximum of ionic conductivities would match with that of cell efficiencies, if the ionic conductivity is a rate determining step in the sol id state DSSCs. However, the maximum composition did not show the maximum solar cell performance, indicating the mismatch between ionic conductivity and cell performance. This suggests that the ionic conductivity may not be the rate controlling step in determining the cell efficiency in these experimental conditions, whereas other parameters such as the electron recombination might play an important role. Thus, we tried to modify the surface of the $TiO_2$ particles by coating a thin metal oxide such as $Al_2O_3$ or $Nb_2O_5$ layer to prevent electron recombination. As a result, the maximum of the cell efficiency was shifted to that of the ionic conductivity. The peak shifts were also attempted to be explained by the diffusion coefficient and the lifetime of electrons in the $TiO_2$ layer.

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A study on the modelling and simulation of robotic assembly cells (로보틱 조립셀의 모델링 및 시뮬레이션에 관한 연구)

  • 홍지민;김대원;이범희;고명삼
    • 제어로봇시스템학회:학술대회논문집
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    • 1990.10a
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    • pp.411-416
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    • 1990
  • A modelling process of a robotic assembly cell and a method for analysis of the assembly cell operation through simulation are presented. An assembly cell including industrial robots is the subject of the model. The states of the assembly cell elements are taken as the state variables and the relationships between the states are described mathematically using the operators. An algorithm for the cell operation is developed from the relationships between the states and the information on the assembly task, and efficient analyses are performed by the simulation results.

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Screening and Isolation of the Cell Differentiation Inducers from Medicinal Plants (I) (생약으로부터 세포분화유도물질의 검색 및 분리 및 분리 (I))

  • Park, Eun-Jung;Kim, Jin-Woong
    • Korean Journal of Pharmacognosy
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    • v.28 no.4
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    • pp.225-232
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    • 1997
  • 300 extracts derived from 100 plants were tested for their potential to induce HL-60 cell differentiation using NBT assay and NSE/SE staining methods. Morphological changes from suspended to adherent state of the cells were also observed by microscopic examination. In result, 55 extracts induced cell differentiation into monocyte/macrophage lineage in the NBT and the NSE assay.

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Temporal and Spatial Regulation of Cell Cycle Genes during Maize Sex Determination (옥수수 성 결정에 있어서 세포주기 유전자들의 시간적, 공간적 조절)

  • Lee, Jung-Ro;Kim, Jong-Cheol
    • Journal of Life Science
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    • v.16 no.5
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    • pp.828-833
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    • 2006
  • Maize (Zea mays L.) pistil cell death and stamen cell arrest are pivotal process on the sex determination, which diverges from bisexual state of floral meristem to unisexual state in staminate or pistillate floret. We investigated the temporal and spatial distribution of cell cycle gene expression during maize sex determination. The positive regulatory genes of cell cycle, cyclin A, cyclin B, cyclin dependent kinase (CDK) and Mad2 were highly expressed in the developing pistil and stamen but the expression was disappeared in the dying pistil and arresting stamens. In contrast, the negative regulatory genes of cell cycle, Wee1 and CDK inhibitor (CKI) were expressed in the arresting stamens in the wild-type ear and tasselseed2 mutant tassel, however, these genes were not detected in dying pistil although the cyclin B gene expression was disappeared. These results suggest that both the pistil cell death and stamen cell arrest process in maize sex determination are involved in cell cycle regulation, but the different expression patterns of negative regulatory cell cycle genes in the arresting stamens and aborting pistils suggest that the two processes may have distinctive modes of action.

Expression Characteristics of Proteins of the Insulin-like Growth Factor Axis in Non-small Cell Lung Cancer Patients with Preexisting Type 2 Diabetes Mellitus

  • Ding, Jing;Tang, Jie;Chen, Xin;Men, Hai-Tao;Luo, Wu-Xia;Du, Yang;Ge, Jun;Li, Cong;Chen, Ye;Cheng, Ke;Qiu, Meng;Liu, Ji-Yan
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.10
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    • pp.5675-5680
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    • 2013
  • Background: Preexisting type 2 diabetes mellitus (T2DM) affects the prognosis and mortality of patients with some cancers. Insulin like growth factor (IGF) and insulin receptor (IR) signaling axes play important roles in both cancer and diabetes development. We aimed to explore the expression characteristics of proteins in IGF/IR axis in non-small cell lung cancer (NSCLC) cases with preexisting T2DM. Methods: Fifty-five NSCLC patients with preexisting T2DM were retrospectively included and matched by 55 NSCLC without diabetes at a 1:1 ratio. The expression of proteins in IGF/IR axis was detected by immunohistochemical staining. Clinicopathological data were collected to analyze their relationship with the protein expression. Results: Both IGF 1 receptor (IGF-1R) and insulin receptor substrate 2 (IRS-2) showed higher expression in the NSCLC with T2DM group, compared with those without T2DM. The high expression of IGF-1R and IRS-2 were found to be negatively associated with lymph node metastases and T staging in the T2DM group, respectively, and IRS-2 expression was also found more in the subgroup whose T2DM duration was more than 4 years. No difference was detected in the expression of IRS-1, IGF-1, IGF-2, IGFBP3, IR and mTOR between groups with or without T2DM. Conclusion: Our study found higher expression of IGF-1R and IRS-2 proteins in NSCLC patients with preexisting T2DM, and that there was an association with early stage NSCLC, which suggested that IGF signaling may play an important early event in development of NSCLC associated with diabetes.

Expression, Purification, and Biological Characterization of The Amino-Terminal Fragment of Urokinase in Pichia pastoris

  • Li, Jianping;Lin, Yuli;Zhuang, Hongqin;Hua, Zi-Chun
    • Journal of Microbiology and Biotechnology
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    • v.23 no.9
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    • pp.1197-1205
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    • 2013
  • Urokinase (uPA) and its receptor (uPAR) play an important role in tumor growth and metastasis. Targeting the excessive activation of this system as well as the proliferation of the tumor vascular endothelial cell would be expected to prevent tumor neovasculature and halt the tumor development. In this regard, the amino-terminal fragment (ATF) of urokinase has been confirmed as effective to inhibit the proliferation, migration, and invasiveness of cancer cells via interrupting the interaction of uPA and uPAR. Previous studies indicated that ATF expressed in Escherichia coli was mainly contained in inclusion bodies and also lacked posttranslational modifications. In this study, the biologically active and soluble ATF was cloned and expressed in Pichia pastoris. The recombinant protein was purified to be homogenous and confirmed to be biologically active. The yield of the active ATF was about 30 mg/l of the P. pastoris culture medium. The recombinant ATF (rATF) could efficiently inhibit angiogenesis, endothelial cell migration, and tumor cell invasion in vitro. Furthermore, it could inhibit in vivo xenograft tumor growth and prolong the survival of tumor-bearing mice significantly by competing with uPA for binding to cell surfaces. Therefore, P. pastoris is a highly efficient and cost-effective expression system for large-scale production of biologically active rATFs for potential therapeutic application.