• Title/Summary/Keyword: cell lysis

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Dual Cytotoxic Responses Induced by Treatment of A549 Human Lung Cancer Cells with Sweet Bee Venom in a Dose-Dependent Manner

  • Yu-Na Hwang;In-Seo Kwon;Han-Heom Na;Jin-Sung Park;Keun-Cheol Kim
    • Journal of Pharmacopuncture
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    • v.25 no.4
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    • pp.390-395
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    • 2022
  • Objectives: Sweet bee venom (sBV) is purified from Apis mellifera, containing a high level of melittin-its main component. It has been used as a therapeutic agent for pain relief and anti-inflammation, as well as for treating neuronal abnormalities. Recently, there have been studies on the therapeutic application of sBV for anticancer treatment. In the present study, we investigated the pharmacological effect of sBV treatment in A549 human lung cancer cells. Methods: We used microscopic analysis to observe the morphological changes in A549 cells after sBV treatment. The MTT assay was used to examine the cytotoxic effect after dose-dependent sBV treatment. Molecular changes in sBV were evaluated by the expression of apoptosis marker proteins using western blot analysis. Results: Microscopic analysis suggested that the growth inhibitory effect occurred in a dose-dependent manner; however, cell lysis occurred at a concentration over 20 ㎍/mL of sBV. The MTT assay indicated that sBV treatment exhibited a growth inhibitory effect at a concentration over 5 ㎍/mL. On fluorescence activated cell sorting analysis, G0 dead cells were observed after G1 arrest at treatment concentrations up to 10 ㎍/mL. However, rapid cell rupture was observed at a concentration of 20 ㎍/mL. Western blot analysis demonstrated that sBV treatment modulated the expression of multiple cell death-related proteins, including cleaved-PARP, cleaved-caspase 9, p53, Bcl2, and Bax. Conclusion: sBV induced cell death in A549 human lung cancer cells at a pharmacological concentration, albeit causing hemolytic cell death at a high concentration.

A Role of Natural Killer Cell in Mouse Infected Herpes Simplex Virus (Herpes Simplex Virus에 감염된 Mouse의 NK세포역할)

  • Lee, Yun-Tai;Lee, Chong-Hoon
    • The Journal of the Korean Society for Microbiology
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    • v.17 no.1
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    • pp.7-14
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    • 1982
  • A model of induction of neoplasia by viruses has develpoed from experimental studies in animals and in cultured cells and oncogenic transformation of cells is the result of integration of viral genetic information into the cellular DNA. The evidence for these associations was derived primarily from seroepidemiologic investigation. However, data indicating that the relation between HSV-2 and cervical cancer fits the model derived from experimental animal studies are not yet sufficient to draw conclusion with regard to the etiologic role the virus in the development of the neoplasms. In other hand, the K562 tumor cell is highly susceptible target for natural killer cell lysis by the lymphocytes of human and murine periperal blood. The characteristics of this effector cell type has been investigated. A study on natural killer cell mediated cytotoxicity(NKMC) against $^{51}Cr$-K562 as target cell was studed in HSV-2 infected ICR mouse. We have studied for susceptibility of HSV-2 against mouse embryo fibroblast(MEF) cells and NKMC from HSV-2 infected mouse. The results obtained that the mouse embryo fibroblast cells culture, the number and size of the cells were markedly increased and formed a monolayers relatively rapid, and become complete monolayer sheet around 72 hrs. Duration of cytopathic effect on MEF cells was rapid by serial passing of HSV-2. The morphology of the HSV-2 infected cells appear to be mainly round, ovium, spindle form and some of them was forming large giant cells. The NKMC was decrease in mouse with HSV-2 and comparison between effector/target cells ratio as 25:1 and 50:1 respectively, the NKMC was found to be more significantly decreased than normal control we have concluded that the natural killer cell activity of the viral infected mouse was shown as a suppressed during the HSV-2 infection, day 7th and 14th.

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Fusarium mangiferae as New Cell Factories for Producing Silver Nanoparticles

  • Hamzah, Haider M.;Salah, Reyam F.;Maroof, Mohammed N.
    • Journal of Microbiology and Biotechnology
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    • v.28 no.10
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    • pp.1654-1663
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    • 2018
  • Finding a safe and broad-spectrum medication is a goal of scientists, pharmacists, and physicians, but developing and fabricating the right medicine can be challenging. The current study describes the formation of silver nanoparticles (AgNPs) by Fusarium mangiferae. It involves the antibiofilm activity of the nanoparticles against Staphylococcus aureus. It also involves cytotoxic effect against mammalian cell lines. Well-dispersed nanoparticles are formed by F. mangiferae. The sizes of the nanoparticles were found to range from 25 to 52 nm, and UV-Vis scan showed absorption around 416-420 nm. SEM, TEM, and AFM results displayed spherical and oval shapes. Furthermore, the FTIR histogram detected amide I and amide II compounds responsible for the stability of AgNPs in an aqueous solution. AgNPs were observed to decrease the formation of biofilm at 75% (v/v). DNA reducing, smearing, and perhaps fragmentation were noticed after treating the bacterial cells with 50% (v/v). Additionally, cell lysis was detected releasing proteins in the supernatant. It was also observed that the AgNPs have the ability to cause 59% cervical cancer cell line (HeLa) deaths at 25% (v/v), however, they showed about 31% toxicity against rat embryo fibroblast transformed cell lines (REF). The results of this study prove the efficiency of AgNPs as an antibiofilm against S. aureus, suggesting that AgNPs could be an alternative to antibiotics. It must also be emphasized that AgNPs displayed cytotoxic behavior against mammalian cell lines. Further studies are needed for assessing risk in relation to the possible benefit of prescribing AgNPs.

Rapid and Simple Method for Isolating Plasmid DNA from Lactic acid Bacteria (유산균 Plasmid DNA의 신속 간편한 분리방법)

  • Bae, Hyung-Seok;Baek, Young-Jin;Kim, Young-Ki;Yoo, Min;Park, Moo-Young
    • Microbiology and Biotechnology Letters
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    • v.13 no.3
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    • pp.289-296
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    • 1985
  • A simple procedure for rapid isolation of plasmid DNA from lactobacillus species and streptococcus species is described. Lactic acid bacteria were cultured in the TCM broth containing 0.5% glycine and plasmid DNA was isolated from cells treated with mutanolysin by alkaline-detergent lysis method. Good results for releasing and isolating plasmid DNA from lactobacillus species were obtained by treatment of cells with 30$\mu\textrm{g}$ of mutanolysin per ml at 37$^{\circ}C$ for 5 to 10 min. For the streptococcus species, the optimum conditions were slightly different. The procedure could be used for rapid characterization of plasmid DNA in Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus helveticus, Streptococcus lactis, Streptococcus faecalis, Streptococcus faecium, and Streptococcus cremoris strains. Using this procedure, plasmids isolated from $1.5m\ell$ cultures could readily be visualized in agarose gel.

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Effect of surface-treatments on flexibility and guided bone regeneration of titanium barrier membrane

  • Kim, Jin-Tae;Kim, Byoung Soo;Jeong, Hee Seok;Heo, Young Ku;Shin, Sang-Wan;Lee, Jeong-Yol;Shim, Young Ho;Lee, Deuk Yong
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.25 no.3
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    • pp.98-104
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    • 2015
  • Titanium barrier membranes are prepared to investigate the effect of surface-treatments, such as machining, electropolishing, anodizing, and electropolishing + TiN coating, on the biocompatibility and physical properties of the membranes. The surface roughness (Ra) of the membrane decreases from machining ($0.37{\pm}0.09{\mu}m$), TiN coating ($0.22{\pm}0.09{\mu}m$), electropolishing ($0.20{\pm}0.03{\mu}m$), to anodizing ($0.15{\pm}0.03{\mu}m$). The highest ductility (24.50 %) is observed for the electropolished Ti membrane. No evidence of causing cell lysis or toxicity is found for the membranes regardless of the surface-treatments. Cell adhesion results of L-929 and MG-63 show that the machined Ti membrane exhibits the highest cell adhesion while the electropolished membrane is the best membrane for the L-929 cell proliferation after 7 days. However, no appreciable difference in MG-63 cell proliferation among variously surface-treated membranes is detected, suggesting that the electropolished Ti membrane is likely to be the best membrane due to the synergic combination of tailored flexibility and excellent fibroblast proliferation.

Protoplast Formation and Regeneration of Bacillus strains producing biopolymer (Biopolymer 생산성 Bacillus속 균주의 원형질체 형성과 재생)

  • Yim, Moo-Hyun;Kim, Seong-Ho
    • Applied Biological Chemistry
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    • v.42 no.1
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    • pp.20-28
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    • 1999
  • To improve Bacillus strains producing biopolymer, conditions for protoplast formation and regeneration were investigated in biopolymer producing Bacillus subtilis K-1 and lactose utilizing Bacillus coagulans. Bacillus subtilis K-1 mutant (SM-2) and Bacillus coagulans mutants (CM-12) were marked auxotrophic and antibiotics-resistant (SM-2) and an antibiotics-resistant mutants, respectively. To formate protoplasts derived from the mutants, conditions were established as follows. For B. subtilis mutant SM-2, its culture in mid-logarithmic phase was added with penicillin G (1.0 unit/ml) and further reacted for 1.5 hr. Cells were collected and then treated in lysis fluid (pH 7.0) containing 0.4 M sucrose and lysozyme $25\;{\mu}g/ml$ for 40 min at $37^{\circ}$. Protoplast formation was very successful (99.6%) and the ratio of cell wall regeneration was 2.4%. For Bacillus coagulans mutant CM-12, its mid-logarithmic phase culture was treated with penicillin G (0.3 unit/ml) and glycine (0.5%) for 1hr. Cells were collected and then resuspended in lysis buffer (pH 7.0) containing 0.6 M lactose and lysozyme $(300\;{\mu}g/ml)$ for 30 min at $37^{\circ}$. Protoplast formation was also successful (90.8%) and cell wall regeneration ratio was similar to SM-2 (2.2%). To improve regeneration frequency, regeneration medium was obtained as followed condition,. Cell wall regeneration was improved 2-4 folds with 5.1% for B. subtilis SM-2 and 10.3% for B. coagulans CM-12 when protoplasts mixed with soft top agar(0.4%) was overlaid onto trypticase soy broth medium containing 0.4 M sucrose, 0.7% casamino acid, 1% PVP, 25 mM $MgCl_2,\;25\;mM\;CaCl_₂$ and 1.5% agar.

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Localization and Function of Cellulase in Endosperm Cells of Panax ginseng Seeds during Maturation and After-ripening (인삼 종자의 성숙과 후숙 과정에서 배유세포내 섬유소 가수분해효소의 분포 및 기능)

  • 유성철
    • Journal of Plant Biology
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    • v.36 no.4
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    • pp.327-335
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    • 1993
  • The active sites, intracellular transport, function of cellulase in association with the disintegration of the storage materials of the endosperm cells during seed maturation and after-ripening of Panax ginseng C.A. Meyer seeds were studied by electron microscopy. Cytochemical activities of the cellulase occurred in protein bodies and vesicles of endosperm cells in seed with red seed coat. In after-ripening seed, the activities were strongly found in the cell wall of endosperm near the umbiliform layer and on neighbouring vesicles, so it is assumed that these cells begin to be decomposed. Cellulase activities were initiated before the decomposition of storage materials. But, no activity was observed in the umbiliform layer, so it is suggested that cellulase lose its activity after the completion of lysis process.

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Occurrence of Saprolegnia diclina Associated with Fungal Gill disease at snake fishes Culture farm (가물치 양식어장(養殖漁場)에서 진균성새병(眞菌性鰓病)과 관련된 Saprolegnia diclina의 발생상황(發生狀況))

  • Min, Hong-Kyu;Chun, Soon-Bai;Bai, Suk
    • Journal of fish pathology
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    • v.4 no.2
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    • pp.95-100
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    • 1991
  • Saprolegnia diclina, which was the pathogen causing death in snake fishes(Channa argus) at culture farm, was investigated using scanning electron microscope. It was found that Saprolegnia diclina infection caused snake fishes to fail gas change in the gills. Cell lysis as well as edematous disease and hyperplasia as a result of Saprolegnea diclina attachment on the surface of gills were observed. The granules, the mean diameters of which ranged from 6 to $7\;{\mu}m$, attaching on the surface of gills were found to be secondary zoospores of Saprolegnia diclina. The failures of gas exchange in the gill cells and circulation as a result of the osmotic dilution of the blood were supposed to be the main cause of death.

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Antiprotozoal Activity of Deacetylated Chitosan Oligosaccharide (dp 2-8) on Trichomonas vaginalis

  • Shin, Woon-Seob;Kil, Jun-Cheul;Park, Gab-Man
    • Journal of Microbiology and Biotechnology
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    • v.16 no.12
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    • pp.1984-1989
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    • 2006
  • Deacetylated chitosan oligosaccharide (COS) had effective antiprotozoal activity against Trichomonas vaginalis (Minimal Inhibitory Concentration, MIC 0.25%), whereas 80% acetylated cas showed no antiprotozoal activity (MIC > 1 %). an the other hand, 80% acetylated cas showed growth stimulatory activity against the protozoa. When T. vaginalis was treated with 98% deacetylated COS at 0.25% concentration, the viability of the protozoa was rapidly decreased within 15 min, and the protozoa completely died within 40 min. Ultrastructural changes of trichomonads treated with COS included a loss of defined nuclear membrane and endoplasmic reticulum membranes, an increase in the number of free ribosome, vacuolation, and ultimately lysis of the cell membrane. These results indicate that deacetylated COS can be used as an antitrichomonal agent, although its lethal mechanism is not known.

Electron Microscopic Studies on the Rat Mast Cells Induced by Morphine Hydrochloride (Morphine Hydrochloride로 인한 흰쥐 장간막 비만세포의 형태학적 변화에 대한 전자현미경적 연구)

  • Kang, Ho-Suck;Kim, Chang-Whan
    • Applied Microscopy
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    • v.3 no.1
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    • pp.39-44
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    • 1973
  • The effects of morphine HCl on the mesenteric mast cells were studied the electron microscopy. The results of the observations are as follows: 1. In the experimental group for intravenous injection of morphine HCl 12 mg/kg, the granules appeared cluster, granular lysis and an electron transparent appearance. Frequently, some granules appeared in the extracellular space. 2. In the experimental group for intravenous injection of morphine HCl 24 mg / kg, it was observed. that the formation of a clear halo or a space around each granule. Many altered granules showing a reticular texture (type 2) are observed in the cytoplasm. 3. From the results mentioned above, it is suggested that rat mesenteric mast cell' granules were affected by morphine HCl.

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