• Korean Journal of Fisheries and Aquatic Sciences
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• v.28 no.4
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• pp.443-450
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• 1995

Listeria monocrogenes has been increasingly recognized as an important food poisoning and pathogenic bacterium which is Gram positive, non spore forming and facultative anaerobic rod shape. Bacteriological and physiological characterization of L. monocytogenes YM-7 isolated from a patient was performed. Optimum growth condition of 1. monocytogenes YM-7 was at $37^{\circ}$, pH 8.0 and $0\%$ sodium chloride in tryptic soy broth, and then it grew pretty well in the range of $8-40^{\circ}C,\;pH 5.0-10.0\;and\;up\;to\;7\%$ of sodium chloride in the medium. The highest hemolysin activity of hemolysin produced by L. monocytogenes YM-7 was shown in the stationary phase of its growth. Hemolysin produced by the isolated strain was stable at $4^{\circ}C\;and\;pH\;6.0-8.0, while it was gradually unstable by increasing the storage temperature.

• Journal of fish pathology
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• v.18 no.2
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• pp.179-185
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• 2005

In Vitro hemolysis and methemoglobin (MetHb) formation in olive flounder rythrocytes were investigated using potassium permanganate ($KMnO_4$) ranging from 2 to 250 ppm, stabilized chlorine dioxide ($S-ClO_2$)ranging from 3.13 to 400 ppm, formalin (37% formaldehyde) ranging from 31.3 to 2,000 ppm and copper sulphate ($CuSO_4$) ranging from 0.04 to 5 ppm. Remarkable hemolysis was found to be induced at $KMnO_4$ concentrations of 31.3-250 ppm and $CuSO_4$ concentrations of 0.63-5 ppm. On the other hand, MetHb formation could not be found at the same treatment concentrations. It is suggested that the cell-damaging system of $KMnO_4$ may be similar from that of $CuSO_4$ in the erythrocytes of olive flounder. Remarkable hemolysis and MetHb formation were found to be induced at $S-ClO_4$ concentrations of more than 25 ppm and 6.25 ppm, respectively. Only $S-ClO_2$ showed both hemolysis and MetHb formation among the chemicals used in the present study. Formalin did not provoke hemolysis at the highest concentration of 2,000 ppm but induced MetHb formation at ranging from 250 to 2,000 ppm. These findings reveal that the mechanism involved in formalin-induced cell-damaging effects differs from that induced by $S-ClO_2$ to olive flounder erythrocytes compared with $KMnO_4$ and $CuSO_4$.

• Applied Biological Chemistry
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• v.49 no.4
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• pp.281-286
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• 2006

Tolaasin, a pore-forming toxin, is a 1,985 Da peptide produced by Pseudomonas tolaasii and causes a brown blotch disease on cultivated mushrooms. Tolaasin forms pores on the plasma membrane of various cells including fungi, bacteria, plant as well as erythrocytes, and destroys cell structure. $Zn^{+2}$ has been known to block the tolaasin activity by an unknown mechanism. Thus, we investigated the inhibitory effects of $Zn^{+2}$ on the tolaasin-induced hemolysis to understand the molecular mechanism of tolaasin-induced pore formation. $Zn^{+2}$ and $Cd^{+2}$ inhibited the tolaasin-induced hemolysis in a dose-dependent manner and their Ki values were 170 ${\mu}M$ and 20 mM, respectively. The effect of $Zn^{+2}$ was reversible since the subsequent addition of EDTA chelates $Zn^{+2}$ and removes the inhibitory effect of $Zn^{+2}$. When an osmotic protectant, PEG 2000, was added, the tolaasin-induced hemolysis was not observed. After the removal of osmotic protectant by centrifugation, resuspended erythrocytes with fresh medium were immediately hemolyzed, while the addition of $Zn^{+2}$ prevented from hemolysis, implying that tolaasin-induced pores on the membrane were already formed in the medium containing osmotic protectant. These results suggest that $Zn^{+2}$ inhibits the activity of tolaasin pores and it has minor effects on the membrane binding of tolaasin and the formation of pore.

• Journal of Applied Biological Chemistry
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• v.61 no.3
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• pp.213-217
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• 2018

Tolaasin secreted by Pseudomonas tolaasii is a peptide toxin and causes brown blotch disease on the cultivated mushrooms by collapsing cellular and fruiting body structure. Toxicity of tolaasin was evaluated by measuring hemolytic activity because tolaasin molecules form membrane pores on the red blood cells and destroy cell membrane structure. In the previous studies, we found that tolaasin cytotoxicity was suppressed by $Zn^{2+}$ and $Ni^{2+}$. $Ni^{2+}$ inhibited the tolaasin-induced hemolysis in a dose-dependent manner and its $K_i$ value was 1.8 mM. The hemolytic activity was completely inhibited at the concentration higher than 10 mM. The inhibitory effect of $Zn^{2+}$ on tolaasin-induced hemolysis was increased in alkaline pH, while that of $Ni^{2+}$was not much dependent on pH. When the pH of buffer solution was increased from pH 7 to pH 9, the time for 50% hemolysis ($T_{50}$) was increased greatly by $100{\mu}M$ $Zn^{2+}$; however, it was slightly increased by 1 mM $Ni^{2+}$ at all pH values. When the synergistic effect of $Zn^{2+}$ and $Ni^{2+}$ on tolaasin-induced hemolysis was measured, it was not dependent on the pH of buffer solution. Molecular elucidation of the difference in pH-dependence of these two metal ions may contribute to understand the mechanism of tolaasin pore formation and cytotoxicity.

• Korean Journal of Clinical Laboratory Science
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• v.56 no.1
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• pp.32-42
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• 2024

This study compared the results of hemolyzed samples and re-collected samples to investigate a hemolysis influence and an acceptable hemolysis index (HI). Before and after hemolysis, alkaline phosphatase (ALP), aspartate aminotransferase (AST), alanine aminotransferase (ALT), amylase (Amy), direct bilirubin (D-bil), total bilirubin (T-bil), creatine phosphokinase (CK), gamma glutamyl transferase (GGT), iron, potassium (K), lactate dehydrogenase (LDH), magnesium (Mg), phosphorus (Phos), total protein (TP), and uric acid (UA) showed significant results in the paired t-test. LDH, K, iron, AST, CK, GGT, TP, Amy and Phos had a high correlation between the degree of hemolysis and the results of samples. When comparing Roche's cut-off HI with HI^QChigh obtained using quality control (QC) high standard deviation (SD), AST, D-bil, CK, and LDH were similar, but Amy, GGT, K, iron, Phos, and TP were lower than the cut-off HI of Roche, while ALP and ALT were higher. Some analytes which showed no significant results in the paired t-test, were found to have significant results in HI>200. Hence, it is suggested that the hemolyzed sample should be rejected if HI>200. Based on this study that some analytes were affected when HI<100, we recommend to set the standard of hemolysis starting from HI>50.

• Applied Biological Chemistry
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• v.50 no.4
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• pp.257-261
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• 2007

Tolaasin, a peptide toxin produced by Pseudomonas tolaasii, causes a serious disease on the cultivated mushrooms, known as brown blotch disease. Hemolysis using red blood cells was designed to measure the cytotoxicity of tolaasin molecules. Since tolaasin has two amine groups near the C-terminus, its membrane binding will be dependent on the ionic states of the amine groups. When the tolaasin peptide was titrated, its titration curve indicated the presence of titratable amine(s) at pH ranges from 7.0 to 9.6. When the pH-dependence of tolaasin-induced hemolysis was measured at various pHs, hemolysis was more efficient at alkaline pHs. In order to measure the membrane binding activity of tolaasin at different pHs, RBCs were incubated with tolaasin molecules for short time periods and washed out with fresh buffer. Because of the tolaasin binding during the preincubation period, fast hemolyses were observed at pH 8 or higher. These results imply that non-charged or less positively charged states of tolaasin molecules easily bind to membrane and show high hemolytic activity.

• The Korean Journal of Food And Nutrition
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• v.10 no.4
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• pp.533-538
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• 1997

This study was conduced to investigate on immune response of the hot water extract(PHE), 50% methanol extract(PME) and acetone extract(PAE) from Glycyrrhiza glabra. The experiment was carried out by phagocytosis, plaque forming cell(PFC), hemalysin titration(HY) and rosette forming cell(RFC) assay by using BALB/c mice. The results obtained from this study are as follow ; The effects of Glycyrrhiza glabra extracts on phagocytosis was tended to be slight increase in GME and GAE groups compared to the control group, but not significant. In the experiment of PFC and HY, the results of experiment groups which was given each samples were significantly higher than the control group. The result of rosette forming cell in GME and GAE groups were significantly higher than control group.

• Journal of Food Hygiene and Safety
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• v.13 no.3
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• pp.232-237
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• 1998

The purpose of this study is to examine the phototoxicity of four phenothiazine derivatives such as chlorpromazine, perphenazine, trifluoroperazine and promethazine, and to investigate the effects of ascorbic acid on their phototoxicity by UVB irradiation. Effects of the test compounds on RBCs were monitored with a spectrophotometer by the method of Kahn et al. The extent of photohemolysis by tested phenothiazine derivatives were increased with their concentration and toxic photoproducts were formed by chlorpromazine and promethazine with preirradiated UVB. Photohemolysis postirradiated chlorpromazine and perphenazine and preirradiated promethazine were decreased with the use of ascorbic acid significantly.

• Microbiology and Biotechnology Letters
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• v.43 no.1
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• pp.84-90
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• 2015

Seventeen ampicillin-sensitive coagulase-negative staphylococci (CNS) isolates identified in jeotgal were subjected to assessments for antibiotic susceptibility and safety hazards. Fifteen of the 17 CNS strains exhibited phenotypic resistances to at least one antibiotic, and their prevailing resistance was to penicillin G. The dfrA gene for trimethoprim and tetK for tetracycline were amplified by PCR from the two strains, respectively. α-Hemolytic activity was not detected from the 17 strains, while five strains presented δ-hemolytic activity. Among the five strains, two strains exhibited β-hemolytic activity. Biofilm was formed from twelve strains. All of the tested phenotypic characteristics were expressed in a strain-specific manner.

• Korean Journal of Microbiology
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• v.35 no.3
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• pp.248-252
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• 1999

The PI-oduction of virulence factors such as cholera toxin, heinolysin and hemagglutinin in V cliolerae non-01 and non-0139 were examined. Among 65 strains isolated from environmental and clinical blood sources, 29 (14.6%) strains produced hemolysin only, 35(53.9%) sh.ains produced both hemolysin and hemagglutinin. From one 037 slrain isolated from environmenl, cholera toxin, ctx gene, hemolysin, and hemagglutinin were detected. All of the strains isolated from clinical and environmental sources showed hemolytic activity against human 0 group e~ythrocytes. In inhibition patterns of heinagglotination, 5 of 18 clinical strains (27.8%) were inhibited by less than 1% mannose and galactose, while, among the 47 environmental isolates. hose paltems by less than 1% mannose and galactose 55.4% wel-e inhibited. Thel-ehre, exohamagglutinin positive rate was high in clinical blood isolates but in environnlental sources, the rate was almost similar lo ihe rate or endohemagglutinin positive. These results indicaled that V cholerae non-01 and non-0139 produced various virnlence factors such as cholera toxin, hemolysin, and hemagglutinin but not a single factor. Further studies are need for epidemiological or bacteriological shtdies of V cholerae 037 isolated from environment.