• Proceedings of the KSAR Conference
• /
• 2002.06a
• /
• pp.37-37
• /
• 2002

The success of embryo cloning depends on numerous factors; interaction between recipient ooplasm and donor nucleus, nuclear reprogramming, oocyte activation, and donor cell cycle and type. In this study, the cell cycle and apoptosis of bovine fetal fibroblast as a donor cell for embryo cloning were evaluated following different activation treatments. (omitted)

• Mycobiology
• /
• v.45 no.1
• /
• pp.25-30
• /
• 2017

Metal-based drugs, such as 1,10-phenanthroline, have demonstrated anticancer, antifungal and antiplasmodium activities. One of the 1,10-phenanthroline derivatives compounds (1)-N-2-methoxybenzyl-1,10-phenanthrolinium bromide (FEN), which has been demonstrated an inhibitory effect on the growth of Candida spp. This study aimed to explore the in vitro antifungal activity of FEN and its effect on the membrane integrity of Candida albicans. The minimum inhibitory concentration (MIC) and the minimum fungicidal concentration (MFC) of FEN against planktonic C. albicans cells were determined using the broth microdilution method according to the Clinical and Laboratory Standards Institute guidelines. Cell membrane integrity was determined with the propidium iodide assay using a flow cytometer and were visualized using scanning electron microscopy (SEM). Planktonic cells growth of C. albicans were inhibited by FEN, with an MIC of $0.39-1.56{\mu}g/mL$ and a MFC that ranged from 3.125 to $100{\mu}g/mL$. When C. albicans was exposed to FEN, the uptake of propidium iodide was increased, which indicated that membrane disruption is the probable mode of action of this compound. There was cells surface changes of C. albicans when observed under SEM.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.22 no.4
• /
• pp.771-777
• /
• 2008

Chungpaesagan-tang which is used for treating patients of brain in cerebrovascular disease frequently from clinical doctor has not reported about the effect of neuronal aptosis caused of brain ischemia. The aim of this study is to investigate effect of Chungpaesagan-tang protecting neuronal cells from being damaged by brain ischemia through using organotypic hippocampal slice cultures. We caused ischemic damage to organotypic hippocampal slice cultures by oxygen and glucose deprivation. And added Chungpaesagan-tang extract to cultures. thereafter we measured area percentage of propidium iodide (PI)-stained neuronal cell, lactate dehydrogenase (LDH) levels in culture media and Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL)-positive cells. Area percentage of PI-stained neuronal cells and count of TUNEL-positive cells in CA1 and DG area of organotypic hippocampal slice culture were significantly decreased in pertinent density level of Chungpaesagan-tang extract. LDH levels in culture media of organotypic hippocampal slice culture were significantly decreased in pertinent density level of Chungpaesagan-tang extract. Within pertinent density level, Chungpaesagan-tang has cell protection effect that prevents brain ischemia damaging neuronal cells and apoptosis increasing.

• Journal of Microbiology and Biotechnology
• /
• v.19 no.12
• /
• pp.1635-1643
• /
• 2009

The aim of this study was to provide new insight into the mechanism whereby the housekeeping enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) locates to cell walls of Lactobacillus plantarum 299v. After purification, cytosolic and cell wall GAPDH (cw-GAPDH) forms were characterized and shown to be identical homotetrameric active enzymes. GAPDH concentration on cell walls was growth-time dependent. Free GAPDH was not observed on the culture supernatant at any time during growth, and provoked cell lysis was not concomitant with any reassociation of GAPDH onto the cell surface. Hence, with the possibility of cw-GAPDH resulting from autolysis being unlikely, entrapment of intracellular GAPDH on the cell wall after a passive efflux through altered plasma membrane was investigated. Flow cytometry was used to assess L. plantarum 299v membrane permeabilization after labeling with propidium iodide (PI). By combining PI uptake and cw-GAPDH activity measurements, we demonstrate here that the increase in cw-GAPDH concentration from the early exponential phase to the late stationary phase is closely related to an increase in plasma membrane permeability during growth. Moreover, we observed that increases in both plasma membrane permeability and cw-GAPDH activity were delayed when glucose was added during L. plantarum 299v growth. Using a double labeling of L. plantarum 299v cells with anti-GAPDH antibodies and propidium iodide, we established unambiguously that cells with impaired membrane manifest five times more cw-GAPDH than unaltered cells. Our results show that plasma membrane permeability appears to be closely related to the efflux of GAPDH on the bacterial cell surface, offering new insight into the understanding of the cell wall location of this enzyme.

• ALGAE
• /
• v.31 no.4
• /
• pp.373-378
• /
• 2016

Cochlodinium polykrikoides is a red-tide forming dinoflagellate that causes significant worldwide impacts on aquaculture industries and the marine ecosystem. There have been extensive studies on managing and preventing C. polykrikoides blooms, but it has been difficult to identify an effective method to control the bloom development. There is also limited genome information on the molecular mechanisms involved in its various ecophysiology and metabolism processes. Thus, comprehensive genome information is required to better understand harmful algal blooms caused by C. polykrikoides. We estimated the C. polykrikoides genome size using flow cytometry, with detection of the fluorescence of DNA stained with propidium iodide (PI). The nuclear genome size of C. polykrikoides was 100.97 Gb, as calculated by comparing its mean fluorescence intensity (MFI) to the MFI of Mus musculus, which is 2.8 Gb. The exceptionally large genome size of C. polykrikoides might indicate its complex physiological and metabolic characteristics. Our optimized protocol for estimating the nuclear genome size of a dinoflagellate using flow cytometry with PI can be applied in studies of other marine organisms.

• The Journal of Internal Korean Medicine
• /
• v.29 no.1
• /
• pp.231-242
• /
• 2008

Objectives : We can find out the experimental reports of Yanggyuksanhwa-tang, which has the function of regulating blood pressure related with cerebral disease, and increasing local cerebral blood stream volume, also has the recoveries for the damage of vessel endothelium, and endothelium hypertrophy caused by angiospasm after subarachnoid hemorrhage, and reduces the contraction of smooth muscle, so simultaneously improves necrosis. The aim of this study is to investigate effect of Yanggyuksanhwa-tang protecting neuronal cells from being damaged by brain ischemia through using organotypic hippocampal slice cultures. Methods : We caused ischemic damage to organotypic hippocampal slice cultures by oxygen and glucose deprivation, and Yanggyuksanhwa-tang extract was added to cultures. Thereafter we measured area percentage of propidium iodide (PI)-stained neuronal cell, lactate dehydrogenase (LDH) levels in culture media and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL)-positive cells. Results : Area percentage of PI-stained neuronal cells and count of TUNEL-positive cells in CA1 and DG area of organotypic hippocampal slice culture were significantly decreased in pertinent density level of Yanggyuksanhwa-tang extract. LDH levels in culture media of organotypic hippocampal slice culture were significantly decreased in pertinent density level of Yanggyuksanhwa-tang extract. Conclusions : Within pertinent density level, Yanggyuksanhwa-tang has cell protection effect that prevents brain ischemia damaging neuronal cells and apoptosis increasing.

• Korean Journal of Microbiology
• /
• v.44 no.4
• /
• pp.299-304
• /
• 2008

Monocytic cells in myeloid lineage are known for latent site of HCMV Previous studies have suggested that HCMV regulates cell cycle progression in a variety of cells, but studies in monocytic cells are limited. In this study, we attempted to understand cell cycle changes after HCMV infection in the monocytic cell lines. Flow cytometric analyses using propidium iodide revealed that the proportion of G0-G1 phase was increased and the proportion of S phase decreased in HCMV-infected THP-1 cells, but not in HL-60 cells. BrdU-incorporation assay supported that cell proliferation was inhibited in HCMV-infected THP-1 cells by inhibition of de novo DNA synthesis. Western blot analysis revealed that p21, inhibitor of cell cycle progression from G1 phase to S phase, was induced in HCMV-infected THP-1 cells but not in HL-60 cells. Thus, HCMV inhibited cell pro-liferation by arresting the cell cycle at G0-G1 phase through induction of p21 protein in promocytic THP-1 cells.

• The Korean Journal of Mycology
• /
• v.29 no.2
• /
• pp.104-109
• /
• 2001

The antifungal effects of polyphosphates on growth of Candida albican and Trichophyton mentagrophytes were studied. The polyphosphates with chain length of 15, 45, and 75 were inhibitory to growth of fungi whereas no inhibition was shown by pyrophosphate. As chain length increase, the more inhibitory effect of the polyphosphates on fungal growth was observed. The concentration of polyphosphate at $800\;{\mu}g/ml$ completely inhibited the growth of fungus. Supplementation of the medium with $Mg^{2+}\;and\;Ca^{2+}$ reduced inhibitory effect of polyphosphate on growth of C. albican treatment of C. albican with polyphosphate, the release of nucleic acid out of cell was observed. When C. albican exposed to polyphosphate were examined, profound changes of cell morphology such as cell swelling and surface blebs were observed. In addition, propidium iodide, membrane impermeable dye, stained the nucleus of C. albican cell treated with polyphosphate. Therefore, it is proposed that the antifungal activity of polyphosphate might be related with its chelation effect to essential cation components of fungal cell wall or membrane.

• Journal of The Korean Ophthalmological Society
• /
• v.59 no.11
• /
• pp.1056-1061
• /
• 2018

Purpose: To investigate the effects of valproic acid on the survival of cultured human Tenon's capsule fibroblasts (HTFBs). Methods: Primary cultured HTFBs were exposed to 0, 0.25, 0.5, and 1.0 mM valproic acid with or without 0, 1.0, $2.5{\mu}g/mL$ mitomycin C, and incubated for 5 days. Cell survival was assessed using an MTT (3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide) assay and the degree of apoptosis was assessed by flow cytometry using annexin-V/propidium iodide double staining. Results: Valproic acid decreased the survival of HTFBs in a dose-dependent manner, and survival was further decreased by adding mitomycin C to valproic acid. Both valproic acid and mitomycin C induced apoptosis of HTFBs. Valproic acid induced less apoptosis than mitomycin C. Conclusions: Valproic acid decreased the cellular survival of HTFBs and induced apoptosis. The antiproliferative effects of valproic acid were further enhanced by the addition of mitomycin C.

• Animal Bioscience
• /
• v.34 no.9
• /
• pp.1525-1531
• /
• 2021

Objective: The objective of this study was to evaluate the antimicrobial effects of fermented Maillard reaction products made by milk proteins (FMRPs) on Clostridium perfringens (C. perfringens), and to elucidate antimicrobial modes of FMRPs on the bacteria, using physiological and morphological analyses. Methods: Antimicrobial effects of FMRPs (whey protein plus galactose fermented by Lactobacillus rhamnosus [L. rhamnosus] 4B15 [Gal-4B15] or Lactobacillus gasseri 4M13 [Gal-4M13], and whey protein plus glucose fermented by L. rhamnosus 4B15 [Glc-4B15] or L. gasseri 4M13 [Glc-4M13]) on C. perfringens were tested by examining growth responses of the pathogen. Iron chelation activity analysis, propidium iodide uptake assay, and morphological analysis with field emission scanning electron microscope (FE-SEM) were conducted to elucidate the modes of antimicrobial activities of FMRPs. Results: When C. perfringens were exposed to the FMRPs, C. perfringens cell counts were decreased (p<0.05) by the all tested FMRPs; iron chelation activities by FMRPs, except for Glc-4M13. Propidium iodide uptake assay indicate that bacterial cellular damage increased in all FMRPs-treated C. perfringens, and it was observed by FE-SEM. Conclusion: These results indicate that the FMRPs can destroy C. perfringens by iron chelation and cell membrane damage. Thus, it could be used in dairy products, and controlling intestinal C. perfringens.