• Applied Microscopy
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• v.19 no.1
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• pp.59-69
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• 1989

Ultrastructure of the secretory portion of the small ampullate gland in the orb web spider, Nephila clavata, has been investigated using the electron microscope. The secretory portion of this gland comprise two different regions which are a S-shaped storage sac and a long, convoluted tail. By the internal textures of the secretory granules, the sac is subdivided into two regions ; the proximal region near the tail and the distal region near the duct. Commonly single layered connectives cover the basal portion of the sac epithelium, and apical portion of the epithelial cells is occupied by the thick cuticles. Within the epithelial cells of both the proximal and distal region, several types of the secretory granules surrounded by a limiting membrane and had characteristic crystalloid are scattered throughout the cytoplasm. The granular size and its electron densities are not coincide with each other according to the maturation level of the granules. The wall of the tail is composed of single layer of columnar epithelial cells, and their nuclei are found at the basal portion of the cells. Dissimilar to the epithelial cells of the sac, apical cuticles are not found at this portion. In the cytoplasm of these cells, numerous secretory granules, synthesized from the rough endoplasmic reticula commonly and had fine fibrous materials, are found. At the cell surface bordering the lumen, microvilli are seen, arid along the cellular boundaries specialized septate junctions and desmosomes appeared.

• The Transactions of The Korean Institute of Electrical Engineers
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• v.60 no.1
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• pp.100-104
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• 2011

In this work, an optimum condition of electrolytes preparation for photovoltaic cells application was investigated experimentally in terms of impedance and conversion efficiency of the cells. 3-methoxyppropionitrie and redox pairs with LiI and $I_2$ were used as stable solvents for fabrication of electrolyte. Efficiency comparison of the prepared cells carried out for various additives and ionic liquids. From the results, there was an optimum concentration (about 0.3 M) of ionic liquids for efficient cell fabrication. For case of electrolyte using single DMAp additive, the maximum conversion efficiency of the cell was 6.4%($V_{oc}$: 0.78V, $J_{sc}$: 14.4 mA/$cm^2$, ff: 0.57). For case of electrolyte using both DMAp and CEMim additives, the maximum conversion efficiency of the cell was 7.2%($V_{oc}$: 0.79V, $J_{sc}$: 16 mA/$cm^2$, ff: 0.57). From the result of electrochemical impedance measurement, both Z1 and Z3 values of binary additives-based cell decreased compared to those of single additive-based. This is due to the decreased in internal and charge transfer resistivities of the cells.

• Journal of fish pathology
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• v.34 no.2
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• pp.177-184
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• 2021

Vaccines based on single-cycle viruses that are replication-incompetent due to knockout of replication-related structural gene(s) are more immunogenic than inactivated or subunit vaccines and can be used as delivery vehicles for foreign antigens without concerns on the reverting to virulent forms. The aim of this study was to develop a delivery vehicle for nervous necrosis virus (NNV)-like particles (VLPs) using G gene deleted single-cycle VHSV (rVHSV-𝚫G). Recombinant single-cycle VHSVs carrying NNV capsid protein gene between N and P gene of rVHSV-𝚫G genome (rVHSV-𝚫G-NNV_Cap) were rescued by reverse genetic technology. The successful expression of NNV capsid protein in cells infected with rVHSV-𝚫G-NNV_Cap was demonstrated by Western blot analysis, and the production of NNV VLPs in infected cells was confirmed using an electron microscopy. The results suggest that single-cycle VHSVs can be used as a safe delivery vehicle for NNV VLPs, and can be extended to other pathogens for the development of prophylactic vaccines.

• The Korean Journal of Physiology and Pharmacology
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• v.3 no.4
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• pp.439-445
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• 1999

This study is to investigate the mechanism of inhibitory effect of imipramine on the calcium utilization in single cells isolated from canine detrusor. 2 mm thick smooth muscle chops were incubated in 0.12% collagenase solution at $36^{circ}C,$ and aerated with 95% $O_2/5%\;CO_2,$ and then cell suspension was examined. Acetylcholine (ACh) evoked a concentration-dependent contraction of the isolated detrusor cells in normal physiologic salt solution (PSS), and the ACh-induced contraction was significantly inhibited by imipramine. In $Ca^{2+}-free$ PSS, ACh-induced contraction was less than those in normal PSS and it was not affected by the pretreatment with imipramine. $Ca^{2+}-induced$ contraction in $Ca^{2+}-free$ PSS was supressed by imipramine, but addition of A 23187, a calcium ionophore, overcomed the inhibitory effect of imipramine. High potassium-depolarization (40 mM KCl) evoked cell contraction, which was inhibited by imipramine. Caffeine, a releasing agent of the stored $Ca^{2+}$ from sarcoplasmic reticulum, evoked a contraction of the cells that was not blocked by the pretreatment with imipramine. These results suggest that imipramine inhibits the influx of calcium in the detrusor cells through both the receptor-operated- and voltage-gated-calcium channels, but does not affect the release of calcium from intracellular storage site.

• The Korean Journal of Physiology and Pharmacology
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• v.13 no.6
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• pp.455-460
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• 2009

Glutamate-induced cobalt uptake reveals that non-NMDA glutamate receptors (GluRs) are present in rat taste bud cells. Previous studies involving glutamate induced cobalt staining suggest this uptake mainly occurs via kainate type GluRs. It is not known which of the 4 types of taste bud cells express subunits of kainate GluR. Circumvallate and foliate papillae of Sprague-Dawley rats (45~60 days old) were used to search for the mRNAs of subunits of non-NMDA GluRs using RT-PCR with specific primers for GluR1-7, KA1 and KA2. We also performed RT-PCR for GluR5, KA1, $PLC\beta2$, and NCAM/SNAP 25 in isolated single cells from taste buds. Taste epithelium, including circumvallate or foliate papilla, express mRNAs of GluR5 and KA1. However, non-taste tongue epithelium expresses no subunits of non-NMDA GluRs. Isolated single cell RT-PCR reveals that the mRNAs of GluR5 and KA1 are preferentially expressed in Type II and Type III cells over Type I cells.

• Toxicological Research
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• v.20 no.3
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• pp.225-232
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• 2004

The suitability of the single cell gel electrophoresis (SCGE) assay as a test for the monitoring of genotoxicity of aquatic environment was evaluated. The SCGE assay was employed to detect DNA damage induced in clam (Spisula sachalinensis) exposed to a direct mutagen, N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) or an indirect mutagen, benzo[a]pyrene (B[a]P). The cells of gill and digestive glands were isolated from clam by homogenization, which was the optimized cell dissociation method, and the level of DNA damage was assessed and expressed as mean tail length. In the gill cells, significant dose- and time-dependent increase was observed in the mean tail length at the concentration from 0.01 to 0.5 ppm MNNG for 96 h. The linear correlation between relative dam-age index (RDI) values was suggested to provide criteria of genotoxicity monitoring for direct acting mutagen. The dose- and time-dependent responses of the digestive glands cells were less sensitive than those of the gill cells. In contrast, the genotoxic response resulting from the exposure of 0.01～1.0 ppm B[a]P to clam revealed a higher sensitivity in the digestive glands cells than the gill cells. The comparison between the time profiles of genotoxic responses in clam and carp, the latter had been obtained in our previous study, indicated that the metabolism of genotoxic compounds in the two aquatic organisms were quite different each other. We conclude that the SCGE assay has the potential as a screening test for routine genotoxicity monitoring of aquatic organisms because of its higher sensitivity and simplicity.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.7
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• pp.1029-1036
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• 2017

Objective: In the livestock industry, the regulatory mechanisms of muscle proliferation and differentiation can be applied to improve traits such as growth and meat production. We investigated the regulatory pathway of MyoD and its role in muscle differentiation in quail myoblast cells. Methods: The MyoD gene was mutated by the clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 technology and single cell-derived MyoD mutant sublines were identified to investigate the global regulatory mechanism responsible for muscle differentiation. Results: The mutation efficiency was 73.3% in the mixed population, and from this population we were able to establish two QM7 MyoD knockout subline (MyoD KO QM7#4) through single cell pick-up and expansion. In the undifferentiated condition, paired box 7 expression in MyoD KO QM7#4 cells was not significantly different from regular QM7 (rQM7) cells. During differentiation, however, myotube formation was dramatically repressed in MyoD KO QM7#4 cells. Moreover, myogenic differentiation-specific transcripts and proteins were not expressed in MyoD KO QM7#4 cells even after an extended differentiation period. These results indicate that MyoD is critical for muscle differentiation. Furthermore, we analyzed the global regulatory interactions by RNA sequencing during muscle differentiation. Conclusion: With CRISPR/Cas9-mediated genomic editing, single cell-derived sublines with a specific knockout gene can be adapted to various aspects of basic research as well as in functional genomics studies.

• Genomics & Informatics
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• v.21 no.2
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• pp.18.1-18.11
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• 2023

Immunologists have activated T cells in vitro using various stimulation methods, including phorbol myristate acetate (PMA)/ionomycin and αCD3/αCD28 agonistic antibodies. PMA stimulates protein kinase C, activating nuclear factor-κB, and ionomycin increases intracellular calcium levels, resulting in activation of nuclear factor of activated T cell. In contrast, αCD3/αCD28 agonistic antibodies activate T cells through ZAP-70, which phosphorylates linker for activation of T cell and SH2-domain-containing leukocyte protein of 76 kD. However, despite the use of these two different in vitro T cell activation methods for decades, the differential effects of chemical-based and antibody-based activation of primary human T cells have not yet been comprehensively described. Using single-cell RNA sequencing (scRNA-seq) technologies to analyze gene expression unbiasedly at the single-cell level, we compared the transcriptomic profiles of the non-physiological and physiological activation methods on human peripheral blood mononuclear cell-derived T cells from four independent donors. Remarkable transcriptomic differences in the expression of cytokines and their respective receptors were identified. We also identified activated CD4 T cell subsets (CD55⁺) enriched specifically by PMA/ionomycin activation. We believe this activated human T cell transcriptome atlas derived from two different activation methods will enhance our understanding, highlight the optimal use of these two in vitro T cell activation assays, and be applied as a reference standard when analyzing activated specific disease-originated T cells through scRNA-seq.

• Korean Journal of Food Science and Technology
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• v.36 no.1
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• pp.58-63
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• 2004

Cell-separating enzymes were used to investigate enzymatic maceration of watermelon and muskmelon containing single cells. Watermelon and muskmelon were macerated with Macerozyme A and Sumyzyme MC for 30-120min. Changes in maceration properties such as yields, color, viscosity, total sugar, total pectin, total polyphenol, particle size distribution, minerals, and free amino acids of suspensions after enzymatic disintegration were investigated. Contents of biochemical components in the supernatant of suspensions increased with increasing treatment time. Suspensions containing single cells showed good thermal stability without affecting original qualities. Mineral content of single-cell suspension was higher than those of watermelon and muskmelon juices. Potassium contents of single-cell suspension and juice were 240.8 and 172.7 mg%, respectively. Results suggest single-cell suspensions of watermelon and muskmelon can he utilized as ingredients for new beverages.

• Korean Journal of Food Science and Technology
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• v.36 no.1
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• pp.64-68
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• 2004

Cell-separating enzyme (Sumyzyme MC) was used to investigate enzymatic maceration of strawberry, sweet persimmon, kiwi, onion, garlic, and cucumber, Maceration rate, volume, brix, color, particle size distribution, and viscosity were determined, and microscopic observation made on suspensions containing single cells. Sweet persimmon and strawberry showed over 90% meceration rates, and kiwi showed 80%. Color, storage test, and sensory evaluations of single-cell suspensions and their filtrates were performed before and after sterilization. Total dietary fiber contents of raw material and single-cell suspension of garlic were 30.77 and 18.55%, respectively, Results indicate fruit and vegetable suspensions produced through enzymatic disintegration using cell-separating enzyme can be utilized as basic materials in the manufacture of single-cell foods.