• Title/Summary/Keyword: single cells

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Ultrastructural Characteristics of Necrosis and Stunt Disease in Red Pepper by the Mixed Infections of Tobacco mosaic virus-U1 or Pepper mild mottle virus and Pepper mottle virus

  • Kim, Dae-Hyun;Cho, Jeom-Deog;Kim, Jae-Hyun;Kim, Jenog-Soo;Cho, Eui-Kyoo
    • The Plant Pathology Journal
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    • v.21 no.3
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    • pp.252-257
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    • 2005
  • In single infection of Tobacco mosaic virus-U1 (TMV­U1) or Pepper mild mottle virus (PMMoV), mosaic symptoms were produced on the chili pepper cultivars of 'Cheongyang' and 'Wangshilgun'. However, in cultivars of 'Manitta' and 'Bugang', no symptoms were occurred. In single infection of Pepper mottle virus (PepMoV), symptoms of mottle and malformation were produced on the tested cultivars of 'Manitta', 'Bugang', 'Cheongyang', and 'Wangshilgun'. In the cultivars of 'Cheongyang' and 'Wangshilgun', synergistic symptoms of stunt and lethal death were induced by mixed infections in the two combinations of TMV-U1 + PepMoV and PMMoV+PepMoV. However, in cultivars of 'Manitta' and 'Bugang', synergistic symptoms were not noted, but mottling which was milder than that of single infection was produced. Cells infected singly with TMV-U1 and PMMoV in the cultivars of 'Cheongyang' and 'Wangshilgun', respectively, had the typical ultra-structures of tobamovirus as the stacked-band structure and multiple spiral aggregate (SA). In the cells and tissues infected with PepMoV on the cultivars of 'Cheongyang', 'Wangshilgun', 'Manitta' and 'Bugang', the potyvirus inclusions of pinwheels, scrolls, lamminated aggregates and amorphous inclusion were observed. In the cells infected mixedly with combinations of TMV­U1+PepMoV and PMMoV+PepMoV, the virus particles and inclusions of the two different viruses were found simultaneously in the same cytoplasm. The amounts of virus particles in mixed infections were more abundant than in single infection. The angled-layer aggregates (ALA) were observed only in the cells infected with both TMV-U1 and PepMoV.

Effect of Propofol, an Intravenous Anesthetic Agent, on $K_{ATP}$ Channels of Pancreatic ${\beta}-cells$ in Rats

  • Park, Eun-Jee;Song, Dae-Kyu;Cheun, Jae-Kyu;Bae, Jung-In;Ho, Won-Kyung;Earm, Yung-E
    • The Korean Journal of Physiology and Pharmacology
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    • v.4 no.1
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    • pp.25-31
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    • 2000
  • ATP-sensitive potassium channels ($K_{ATP}$ channels) play an important role in insulin secretion from pancreatic beta cells. We have investigated the effect of propofol on $K_{ATP}$ channels in cultured single pancreatic beta cells of rats. Channel activity was recorded from membrane patches using the patch-clamp technique. In the inside-out configuration bath-applied propofol inhibited the $K_{ATP}$ channel activities in a dose-dependent manner. The half-maximal inhibition dose (ED50) was $48.6{\pm}8.4\;{\mu}M$ and the Hill coefficient was $0.73{\pm}0.11.$ Single channel conductance calculated from the slope of the relationship between single channel current and pipette potential $(+20{\sim}+100\;mV)$ was not significantly altered by propofol $(control:\;60.0{\pm}2.7\;pS,\;0.1\;mM\;propofol:\;58.7{\pm}3.5\;pS).$ However, mean closed time was surely increased. Above results indicate that propofol blocks the $K_{ATP}$ channels in the pancreatic beta cells in the range of its blood concentrations during anesthesia, suggesting a possible effect on insulin secretion and blood glucose level.

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Trophoblast Cell Subtypes and Dysfunction in the Placenta of Individuals with Preeclampsia Revealed by Single-Cell RNA Sequencing

  • Zhou, Wenbo;Wang, Huiyan;Yang, Yuqi;Guo, Fang;Yu, Bin;Su, Zhaoliang
    • Molecules and Cells
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    • v.45 no.5
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    • pp.317-328
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    • 2022
  • Trophoblasts, important functional cells in the placenta, play a critical role in maintaining placental function. The heterogeneity of trophoblasts has been reported, but little is known about the trophoblast subtypes and distinctive functions during preeclampsia (PE). In this study, we aimed to gain insight into the cell type-specific transcriptomic changes by performing unbiased single-cell RNA sequencing (scRNA-seq) of placental tissue samples, including those of patients diagnosed with PE and matched healthy controls. A total of 29,006 cells were identified in 11 cell types, including trophoblasts and immune cells, and the functions of the trophoblast subtypes in the PE group and the control group were also analyzed. As an important trophoblast subtype, extravillous trophoblasts (EVTs) were further divided into 4 subgroups, and their functions were preliminarily analyzed. We found that some biological processes related to pregnancy, hormone secretion and immunity changed in the PE group. We also identified and analyzed the regulatory network of transcription factors (TFs) identified in the EVTs, among which 3 modules were decreased in the PE group. Then, through in vitro cell experiments, we found that in one of the modules, CEBPB and GTF2B may be involved in EVT dysfunction in PE. In conclusion, our study showed the different transcriptional profiles and regulatory modules in trophoblasts between placentas in the control and PE groups at the single-cell level; these changes may be involved in the pathological process of PE, providing a new molecular theoretical basis for preeclamptic trophoblast dysfunction.

Studies on Multi-step Addition of NMP in (LiNi0.80Co0.15Al0.05) (NCA) Cathode Slurry Preparation and its Rheological, Mechanical Strength and Electrochemical Properties for Li-ion Cells

  • Vasudevarao Pasala;Satyanarayana Maddukuri;V. Sethuraman;Rekha Lankipalli;Devi Gajula;Venkateswarlu Manne
    • Journal of Electrochemical Science and Technology
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    • v.14 no.3
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    • pp.262-271
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    • 2023
  • For electrode stability and the electrochemical performance of the Li-ion cell, it is essential that the active ingredients and unique additives in the polymer binder be well dispersed with the solvent-based slurry. The efficient procedure used to create the slurry affects the rheological characteristics of the electrode slurry. When successively adding different steps of Nmethyl-2-pyrrolidone (NMP) solvent to the cathode composition, it is evenly disseminated. The electrochemical performance of the Li-ion cells and the electrodes made with slurry formed by single step and multiple steps of addition of NMP solvent are examined. To preform rheological properties of cathode electrode slurry on Ni-rich Lithium Nickel-Cobalt-Aluminum Oxide (LiNi0.80Co0.15Al0.05) (NCA). Also, we investigate different step addition of electrode formation and mechanical strength characterization like peel strength. According to the EIS study, a multi-step electrode slurry has lower internal resistance than a single-step electrode slurry, which results in better electrical characteristics and efficiency. Further, microstructure of electrodes is obtained electrochemical performance in the 18650 cylindrical cells with targeted capacity of 1.5 Ah. The slurry of electrodes prepared by single step and multiple steps of addition of NMP solvent and its effect on the fabrication of 1.5 Ah cells. A three-step solvent addition on slurry has been found to be a lower internal resistance than a single-step electrode slurry as confirmed by the EIS analysis, yielding improved electrical properties and efficiency.

Isolation and Characterization of Mammary FpithelialStem Cells in Culture (유선상피 간세포의 분리 및 특성연구)

  • ;;Kelly H. Clifton
    • Journal of Life Science
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    • v.10 no.1
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    • pp.37-44
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    • 2000
  • The mammary gland contains a subpopulation of epithelial cells with large proliferative potentials which are the likely targets for carcinogens. These clonogenic cells can proliferate and differentiate into functional glandular structures. Rat mammary epithelial cells (RMEC) were isolated and characterized in vitro. By flow cytometry of RMEC stained with fluorescein isothiocyanate-peanut agglutinin(PNA) and phycoerythrin anti-Thy-1.1 monoclonal antibody, it was possible to four cell subpopulations from 7-8 week old F344 female rat mammary glands: cells negative to both reagents (B-), PNA-positive cells (PNA+), Thy-1.1-positive cells (Thy-1.1+), and cells positive to both reagents (B+). When single PNA+ cells were isolated and cultured in Matrigel with irradiated (∼50 Gray) 3T3 fibroblast feeder layer, they gave rise to multicellular clonal structures of three types: alveolar, foamy alveolar, and squamous colonies. The developed structures were similar to the mammary glands in vivo. These results suggest that some of PNA+ cells possesses many of the characteristics of multipotent clonogenic stem-like cells.

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Impact of Co-transfection with Livin and Survivin shRNA Expression Vectors on Biological Behavior of HepG2 Cells

  • Xu, Wei;Chang, Hong;Qin, Cheng-Kun;Zhai, Yun-Peng
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.9
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    • pp.5467-5472
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    • 2013
  • Objective: To construct short hairpin RNA (shRNA) eukaryotic expression vectors targeting Livin and Survivin genes, and to explore the impact of co-transfection of Livin and Survivin shRNA expression vectors on the biological behavior of HepG2 cells. Methods: shRNA eukaryotic expression vectors pSD11-Livin and pSD11-Survivin were designed and constructed then transfected into HepG2 cells separately or in combination. mRNA and protein expression in transfected cells was assessed by quantitative fluorescence PCR and Western blotting, respectively. Cell proliferation was measured by MTT assay and cell apoptosis by TUNEL assay. Results: The Livin and Survivin shRNA eukaryotic expression vectors were successfully constructed and transfected into HepG2 cells. The relative mRNA expression levels of Livin and Survivin in HepG2 cells co-transfected with pSD11-Livin and pSD11-Survivin were $0.12{\pm}0.02$ and $0.33{\pm}0.13$, respectively, which was significantly lower than levels in cells transfected with either pSD11-Livin or pSD11-Survivin (P<0.05). The relative protein expression levels of Livin and Survivin in the co-transfected cells were also significantly decreased compared to single-transfection (P<0.05). The inhibition rate of cell growth in the co-transfection group was higher than that in the single-transfection groups at 48 h, 60 h, or 72 h after transfection (P<0.01). The apoptotic rate increased to the greatest extent in the co-transfection group relative to any other group (P<0.05). Conclusions: Co-transfection with pSD11-Livin and pSD11-Survivin was more efficient than transfection with either vector alone in reducing the mRNA and protein expression of Livin and Survivin genes in HepG2 cells. Co-transfection also inhibited the proliferation of transfected cells more than the other groups, and induced cellular apoptosis more effectively.

Ultrastructure of Endocrine Cells in the Midgut Epithelium of the Japanese Cockroach, Periplaneta japonica (집바퀴의 중장 상피조직내에 있는 내분비세포의 미세구조)

  • Yu, Chai-Hyeock
    • Applied Microscopy
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    • v.26 no.4
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    • pp.421-430
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    • 1996
  • The endocrine cells in the midgut epithelium of the Japanese cockroach, Periplaneta japonica were observed by the light and electron microscopy. The midgut epithelium of the last instar larva and adult cockroach consisted of principal columnar cells, regenerative cells, and secretory granular cells. Midgut endocrine cells were positioned basally as a cone-shaped single cell in the epithelium or underneath the regenerative crypt cells. When midgut epithelium grows and the cell composing it transforms, between the endocrine cells and regenerative cells were made desmosome type junction and large vesicular shaped stretches of loose contact. The endocrine cells were characterized by a clear cytoplasm with abundant Golgi complex and numerous secretory granules. The secretory granules in the cell were spherical and electron dense with their diameter of $200{\sim}400nm$. The secretory granules have been observed as discharged by exocytosis on the basal and lateral side of the cell.

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Comparison between Single and Co-culture of Adipocyte and Muscle Cell Lines in Cell Morphology and Cytosolic Substances (지방과 근육 세포주의 단독 및 공동배양을 통한 세포형태학 및 세포물질 비교 연구)

  • Choi, Chang-Weon;Cho, Won-Mo;Yeon, Seong-Heum;HwangBo, Soon;Song, Man-Kang;Park, Sung-Kwon;Baek, Kyung-Hoon
    • Journal of Animal Science and Technology
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    • v.54 no.2
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    • pp.103-109
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    • 2012
  • Present study was performed to investigate the effect of single and co-culture of adipocyte and muscle cell lines on cell differentiation. 3T3-L1 (adipocyte) and L6 (muscle) cell lines were single-cultured on the condition of 10% fetal bovine serum (FBS)/Dulbeco's modified eagle's medium (DMEM) for 48 h followed by culture within 5% FBS/DMEM as a growth media. Then, the growth media was replaced by differentiation media composed of 2% FBS/DMEM without additives in single- or co-culture of the 3T3-L1 and the L6 cells to induce differentiation of both cell types. In co-culture system, the 3T3-L1 and the L6 cells were grown in separated places by being seeded on a $0.4{\mu}m$ insert membrane and on the bottom of 6 well plate, respectively. Cell differentiation was measured using morphological investigation and cytosolic analysis of glycerol-3-phosphate dehydrogenase (GPDH; for 3T3-L1) and creatine kinase (CK; for L6). Based on the GPDH results, the presence of L6 cells did not stimulate 3T3-L1 differentiation showing more differentiation of 3T3-L1 cells in the single-culture compared to the co-culture condition. In contrast, 3T3-L1 cells in the co-culture promoted differentiation of L6 cells. Enzymatic analysis supported this result showing that 3T3-L1 cells showed statistically (P<0.05) higher GPDH activity in the single-culture than the co-culture, whereas CK results of L6 cells were vice versa (P<0.05). Overall, present results may indicate that co-culture system is more reliable and precise technique compared to single-culture. Further studies on several co-culture trials including different media conditions, supplementation of differentiating substances, molecular biological analysis, etc. should be required to obtain practical and fundamental mass data.

Isolation and Characterization of Single-Chain Fv Against Ductal Cells

  • Lee Myung-Hoon;Ryu Hye-Myung;Kim Sun-Zoo;Park Ji-Young;Uhm Ji-Hyun;Park Tae-In
    • Biomedical Science Letters
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    • v.10 no.3
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    • pp.211-217
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    • 2004
  • For discrimination of ductal and ascinar cells, we isolated a single-chain variable domain fragment (scFv) antibody against ductal cells of salivary gland using phage display technique. From the spleen of a mouse immunized with ductal cell lysate, total RNA was prepared and used as a template for cDNA synthesis of antibody genes. The scFv genes were constructed with variable domain genes of heavy and light chain and were introduced into pCANTAB5E to construct phage scFv library. The phage particles specific for acinar cells were screened by subtraction using immunotubes coated with acinar and ductal cell lysate and enzyme-linked immunoabsorbance assay (ELISA). The characteristics of the scFv were determined by immunohistochemistry (IHC) and the result indicated that the isolated scFv has the specificity against ductal cells of salivary glands and tubules of kidney. And the scFv has an unique binding activity specific for Hashimoto's thyroiditis. The nucleotide sequence of isolated scFv gene was determined and revealed that V/sub H/ belongs to the mouse H-chain family subgroup IB and V/sub L/ to the mouse L-chain family subgroup III.

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Effects of Higenamine on the Calcium Current and the Action Potential in the Guinea-pig Myocytes (Higenamine이 단일심근세포의 Ca-전류 및 활동전압에 미치는 효과)

  • Kim, Young-Duck;So, In-Suk;Earm, Yung-E
    • The Korean Journal of Physiology
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    • v.21 no.2
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    • pp.169-177
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    • 1987
  • The effects of higenamine were investigated in the single atrial and ventricular myocyte of the guinea pig by using patch clamp method. The results obtained were as follows: 1) Isoprenaline which is known to be ${\beta}-agonist$ increased the duration of action potential and calcium current in ventricular cells. 2) Higenamine also increased the duration of action potential and calcium current in ventricular myocytes. And its effect was blocked by propranolol. 3) In the atrial cells, isoprenaline showed ${\beta}-agonist$ effects, which were increasing the duration of action potential and calcium current same as in ventricular cells. 4) Higenamine, however, showed the opposite effects of ${\beta}-agonist$ which were decreasing the duration of action potential and calcium current. The above results suggest that higenamine has the typical ${\beta}-agonist$ effect in ventricular cells but inhibitory effect in atrial cells and this effect on atrium could be due to the reduction of calcium current.

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