• Proceedings of the KOSOMBE Conference
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• v.1996 no.11
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• pp.357-362
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• 1996

A way of quantitatively describing the tissue architecture we have investigated when developing a computer program for malignancy grading of transitional cell bladder carcinoma. The minimum spanning trees, MST was created by connecting the center points of the nuclei in the tissue section image. These nuclei were found by thresholding the image at an automatically determined threshold followed by a connected component labeling and a watershed algorithm for separation of overlapping nuclei. Clusters were defined in the MST by thresholding the edge lengths. For these clusters geometric and densitometric features were measures. These features were compared by multivariate statistical methods to the subjective grading by the pathologists and the resulting correspondence was 85% on a material of 40 samples.

• BMB Reports
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• v.41 no.10
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• pp.699-704
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• 2008

Embryonic stem (ES) cell-derived cardiomyocytes (ESCMs) must be specifically purified in order to prevent teratoma formation, and this confusing issue has hampered their clinical application. We therefore investigated a technique to generate pure labeled ESCMs for possible use in cardiac repair. We generated transgenic ES cell lines expressing enhanced green fluorescent protein (EGFP) under the transcriptional control of the $\alpha$-cardiac myosin heavy chain ($\alpha$-MHC) promoter. Differentiated EGFP-positive ES cells displayed characteristics of CMs. Furthermore, neuregulin-1 (NRG-1) upregulated the expression of the cardiac-restricted transcription factors Nkx2.5 and GATA-4, as well as differentiated CM factors ($\alpha$-MHC, $\beta$-MHC). Immunohistochemistry demonstrated that NRG-1 increased expression of cardiac-specific troponin T in the beating foci of the embryoid bodies. This work revealed a potential method for specifically labeling and enriching ESCMs by combining genetically-engineered ES cell clones and exogenous growth factor treatment.

• Molecules and Cells
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• v.19 no.1
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• pp.74-80
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• 2005

Stress is known to inhibit granule cell proliferation in the hippocampus. However, recent studies suggest that the commonly used dose of bromodeoxyuridine (BrdU) is insufficient to label all fractions of granule cells. Furthermore, stress-induced changes in BrdU availability may influence the labeling of newly born cells. To investigate whether changes in BrdU availability affect measurements of stress-induced granule cell proliferation, granule cell proliferation was assessed using injection of high doses of BrdU before and after restraint stress lasting 1 h. In addition, to determine whether stress-induced changes in plasma corticosterone levels were influenced by the BrdU, time-dependent changes in plasma corticosterone levels over 2 h after BrdU injection were compared with total accumulated plasma corticosterone levels [as determined by areas under the curve (AUC)]. Restraint stress significantly reduced the numbers of BrdU-labeled cells and clusters in the granule cell layer (GCL) of rats that received BrdU after stress, and decreases of similar magnitude were observed when the rats were given BrdU before stress. BrdU injection enhanced the stress-induced plasma corticosterone response, but there was no difference between the mean AUCs of plasma corticosterone levels of animals injected with BrdU before or after stress. These observations suggest that restraint stress decreases granule cell proliferation, and that this may be influenced by the extent and duration of plasma corticosterone increases rather than by changes in the availability of BrdU.

• The Journal of Korean Medicine
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• v.26 no.4
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• pp.12-21
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• 2005

Objectives: In the present study, we investigated whether an aqueous extract of Armeniacae semen induces apoptotic neuronal cell death upon mouse N2a neuroblastoma cells. Methods: 1. Cell viability was determined by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTI) assay. 2. For in situ detection of apoptotic cells, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay, 4,6-diamidino-2-phenylindole (DAPI) staining. 3. The fraction of cells was revealed by flow cytometric analysis used that. 4. For detection of apoptotic DNA cleavage, DNA fragmentation assay was performed. 5. For detection of bax and bcl-2, Western blot analysis was performed. 6. Caspase enzyme activity was measured using caspase-3 assay. Results: From the present results, N2a neuroblastoma cells treated with Armeniacae semen extract exhibited several characteristics of apoptosis. A treatment of Armeniacae semen extract was shown to increase the expression of Bax, a proapoptotic protein, and the treatment decreased the expression of Blc2, an anti-apoptotic protein. In addition, Armeniacae semen extract increased the caspase-3 enzyme activity. Conclusions: The present results show that Armeniacae semen extract induces apoptotic cell death in mouse N2a neuroblastoma cells.

• Journal of Microbiology and Biotechnology
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• v.14 no.3
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• pp.450-455
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• 2004

A reducing glucose-carrying polymer, called poly [3-O-(4'-vinylbenzyl)-D-glucose］(PVG), was interacted with HepG2 cells including a type-l glucose transporter (GLUT-1) on the cell membrane. The cooperative interaction between a number of GLUT-1s and a number of reducing 3-O-methyl-D-glucose moieties on the PVG polymer chain was found to be responsible for the increase in the interaction with HepG2 cells. The affinity between the cells and the PVG was studied using RITC-labeled glycopolymers. The specific interaction between the GLUT-1 on HepG2 cells and the PVG polymer carrying reducing glucose moieties was suppressed by the inhibitors, phloretin, phloridzin, and cytochalasin B. Direct observation by confocal laser microscopy with the use of RITC-labeled PVG and pretreatment of HepG2 cells with the inhibitors demonstrated that the cells interacted with the soluble form of the PVG polymer via GLUT-1, while fluorescence labeling of the cell surface was prevented after pretreatment with the inhibitors of GLUT-1.

• Natural Product Sciences
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• v.25 no.4
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• pp.298-303
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• 2019

This study investigated the cytotoxic effects and mechanism of action of asiatic acid in pancreatic cancer cell lines. First, we confirmed the cell viability of MIA PaCa-2 and PANC-1 cells after asiatic acid administration for 48 and 72 h. The viability of MIA PaCa-2 and PANC-1 cells decreased in a dose-dependent manner following asiatic acid administration. To investigate the underlying mechanism, we performed a terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, annexin V assay, and western blotting. Asiatic acid induced apoptosis and autophagy through activation of AMP-activated protein kinase (AMPK) and inhibition of mammalian target of rapamycin (mTOR) in MIA PaCa-2 cells. Finally, the expression of miR-17 and miR-21, known as oncogenes in pancreatic cancer, was decreased by asiatic acid. These results indicate that asiatic acid has potential as a new therapeutic agent against pancreatic cancer.

• Animal cells and systems
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• v.5 no.3
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• pp.211-216
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• 2001

Polyclonal antiserum against Manduca sexta allatotropin has been utilized to investigate the localization of allatotropin-immunoreactivity in the brain of the si1k moth Bombyx mori. Manduca sexta allatotropin-immunoreactive (Mas-AT-IR) neurons were found in all larval brains investigated, but not in prepupal, pupal and adult brains. In the larval stages, first appearance of Mas-AT-immunoreactivity w8s shown in the brain of first instar larvae, which contains four pairs of bilateral Mas-AT-IR cell bodies. Labeled neurons increased to six pairs in the second instar larval brain, including two pairs of median neurosecretory cells in the pars intercerebralis. In the third and fourth instar larvae, five pairs of labeled cell bodies were distributed throughout each brain. In the fifth instar, there were about ten pairs of bilateral cell bodies in the day-1 brain, about seven pairs in the day-3 brains, and five pairs in the day-5 brains, respectively. Mas-AT-labeling was observed in both axons within nervi corpora cavdiaci (NCC) 1+11 and corpora allata. This suggests that the Mas-AT produced from the brain neurons is transported via some axons of the NCC 1+11 and nervi corpora allati I to the corpora allata, which appears to be a main accumulation site for the Mas-AT neuropeptide in some brain neurons produced in B. mori.

• The Korean Journal of Physiology and Pharmacology
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• v.4 no.1
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• pp.73-79
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• 2000

This study investigated whether propofol, an intravenous, non-barbiturate anesthetic, could reduce brain damage following global forebrain ischemia. Transient global ischemia was induced in gerbils by occlusion of bilateral carotid arteries for 3 min. Propofol (50 mg/kg) was administered intraperitoneally 30 min before, immediately after, and at 1 h, 2 h, 6 h after occlusion. Thereafter, propofol was administered twice daily for three days. Treated animals were processed in parallel with ischemic animals receiving 10% intralipid as a vehicle or with sham-operated controls. In histologic findings, counts of viable neurons were made in the pyramidal cell layer of the hippocampal CA1 area 4 days after ischemia. The number of viable neurons in the pyramidal cell layer of CA1 area was similar in animals treated with a vehicle or a subanesthetic dose of propofol. In terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end-labeling (TUNEL) assay, semiquantitative analysis of dark-brown neuronal cells was made in the hippocampal CA1 area. There was no significant difference in the degree of TUNEL staining in the hippocampal CA1 area between vehicle-treated and propofol-treated animals. These results show that subanesthetic dose of propofol does not reduce delayed neuronal cell death following transient global ischemia in Mongolian gerbils.

• Journal of Korea Multimedia Society
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• v.24 no.8
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• pp.1000-1011
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• 2021

In recent years, using Deep Learning methods to apply for medical and biomedical image analysis has seen many advancements. In clinical, using Deep Learning-based approaches for cancer image analysis is one of the key applications for cancer detection and treatment. However, the scarcity and shortage of labeling images make the task of cancer detection and analysis difficult to reach high accuracy. In 2015, the Unet model was introduced and gained much attention from researchers in the field. The success of Unet model is the ability to produce high accuracy with very few input images. Since the development of Unet, there are many variants and modifications of Unet related architecture. This paper proposes a new approach of using Unet++ with pretrained EfficientNet as backbone architecture for breast tumor cell nuclei segmentation and uses the multi-organ transfer learning approach to segment nuclei of breast tumor cells. We attempt to experiment and evaluate the performance of the network on the MonuSeg training dataset and Triple Negative Breast Cancer (TNBC) testing dataset, both are Hematoxylin and Eosin (H & E)-stained images. The results have shown that EfficientUnet++ architecture and the multi-organ transfer learning approach had outperformed other techniques and produced notable accuracy for breast tumor cell nuclei segmentation.

• Korean Journal of Microbiology
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• v.21 no.2
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• pp.61-70
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• 1983

Synchronized cat kidney cells chronically infected with feline leukemia virus (FeLV) were used to study virus production, the synthesis of group specific antigen (gag) and envelope (env) proteins, the expression of env protein on the cell surface during the cell cycle, and the stability of viral RNA. As detecting method, we developed the radioimmunoassay (RIA) system using beta-emission of $^{131}I$ and demonstrated the validity of this system by comparison with routine RIA system using gamma-emission of $^{125}I$. The produced virus was analysed by developed RIA interval was determined by measuring reverse transcriptase activity. The results show that infected cells produce the complete virus particle containing products of gag, env and pol genes of FeLV, and maximum virus production occurs during mitosis of synchronized cells. Labeling of the cell surface of synchronized cells with $^{131}I$ shows that the amount of $gp70^{env}$ on the cell surface parallels cellular gorwth. Therefore, the cell cycle-dependent release of virus is not petition RIA of synchronized cells with $^{131}I$ labeled viral proteins synthesis during the cell cycle. The rate of synthesis of gag protein shows three peaks, corresponding to the $G_1,\;late\;S\;and\;late\;G_2$ phases of cell cycle. But the rate of synthesis of env protein dose not change, suggesting that in these cells the synthesis of these two gene products in controlled seperately. In Actionomycin D treated cells, the synthesis of viral proteins decreased sharply from 8 hours after treatment, and the late S and $G_2$ peaks of gag protein synthesis were disappeared. This shows the stability of viral RNA for about 6 hours in the absence of continuing viral RNA synthesis.