• Toxicological Research
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• v.29 no.2
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• pp.99-106
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• 2013

Clostridium difficile infection (CDI) has become a significant threat to public health. Although broad-spectrum antibiotic therapy is the primary treatment option for CDI, its use has evident limitations. Probiotics have been proved to be effective in the treatment of CDI and are a promising therapeutic option for CDI. In this study, 4 strains of lactic acid bacteria (LAB), namely, Lactobacillus rhamnosus (LR5), Lactococcuslactis (SL3), Bifidobacterium breve (BR3), and Bifidobacterium lactis (BL3) were evaluated for their anti-C. difficile activity. Co-culture incubation of C. difficile ($10^6$ and $10^{10}$ CFU/ml) with each strain of LAB indicated that SL3 possessed the highest antimicrobial activity over a 24-hr period. The cell-free supernatants of the 4 LAB strains exhibited $MIC_{50}$ values between 0.424 mg/ml (SL3) and 1.318 (BR3) mg/ml. These results may provide a basis for alternative therapies for the treatment of C. difficile-associated gut disorders.

• Toxicological Research
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• v.28 no.3
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• pp.151-157
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• 2012

This paper describes the effects of thermally oxidized tallow on the serum lipids profile and radical scavenging activity (RSA) of the lipids extracted from the different tissues of the rabbits. Tallow was thermally oxidized at $130^{\circ}C$ for 9, 18, 27, 36 and 45 h respectively. Thermally oxidized tallow was fed to the local strain of Himalayan rabbits for one week. Results show that oxidation increases the formation of hydroperoxides and decrease the level of radical scavenging activity of the tallow. The rabbit serum lipids profile showed a dose dependent increase in triglyceride, total cholesterol and LDL-cholesterol. However, no statistically significant increase was observed in the HDL-cholesterol with an increase of oxidation time. Serum glucose and rabbits body weight decrease significantly (p < 0.05) and was highly correlated with the serum lipids profile. The percent RSA of the lipids extracted from the liver, brain and muscles tissues showed a significant decrease with respect to 0.5, 1.0 and 1.5 g/body weight as well as oxidation time. Data suggests that thermal oxidation and use of thermally oxidized beef tallow is harmful and therefore an alternative way of cooking should be used.

• Bulletin of the Korean Chemical Society
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• v.33 no.12
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• pp.3998-4002
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• 2012

With continuing progress of nanotechnologies and various applications of nanoparticles, one needs to develop a quick and fairly standard assessment tool to evaluate cytotoxicity of nanoparticles. However, much cytotoxicity studies on the interpretation of the interaction between nanoparticles and cells are non-mechanistic and time-consuming. Here, we propose a simple screening method for the analysis of the interaction between several AgNPs (5.3 to 64 nm) and fluorescence-dye containing vesicles ($12{\mu}m$) acting as a biomimetic cell-membrane. Fluorescence-dye containing vesicle was prepared using a fluorescence probe (1,6-diphenyl-1,3,5-hexatryene), which was intercalated into the lipid bilayer due to their hydrophobicity. Zeta potential of all materials except for bare-AgNPs (+32.8 mV) was negative (-26 to -54 mV). The morphological change (i.e., rupture and fusion of vesicle, and release of dye) after mixing of the vesicle and AgNPs was observed by fluorescence microscopy, and fluorescence image were different with coating materials and surface charge of x-AgNPs. In the results, we found that the surface charge of nanoparticles is the key factor for vesicle rupture and fusion. This proposed method might be useful for analyzing the cytotoxicity of nanoparticles with cell-membranes instead of in vitro or in vivo cytotoxicity tests.

• Molecular & Cellular Toxicology
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• v.4 no.2
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• pp.157-163
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• 2008

Formation of ${\beta}$-amyloid $(A{\beta})$ fibrils has been identified as one of the major characteristics of Alzheimer's disease (AD). Inhibition of $A{\beta}$ fibril formation in the CNS would be attractive therapeutic targets for the treatment of AD. Several small compounds that inhibit amyloid formation or amyloid neurotoxicity in vitro have been known. Citrate has surfactant function effect because of its molecular structure having high anionic charge density, in addition to the well-known antibacterial and antioxidant properties. Therefore, we hypothesized that citrate might have the inhibitory effect against $A{\beta}$ fibril formation in vitro and have the protective effect against $A{\beta}$-induced neurotoxicity in PC12 cells. We examined the effect of citrate against the formation of $A{\beta}$ fibrils by measuring the intensity of fluorescence in thioflavin-T (Th-T) assay of between $A{\beta}_{25-35}$ groups treated with citrate and the control with $A{\beta}_{25-35}$ alone. The neuroprotective effect of citrate against $A{\beta}$-induced toxicity in PC12 cells was investigated using the WST-1 assay. Fluorescence spectroscopy showed that citrate inhibited dose-dependently the formation of $A{\beta}$ fibrils from ${\beta}$-amyloid peptides. The inhibition percentages of $A{\beta}$ fibril formation by citrate (1, 2.5, and 5 mM) were 31%, 60%, and 68% at 7 days, respectively in thioflavin-T (Th-T) assay. WST-1 assay revealed that the toxic effect of $A{\beta}_{25-35}$ was reduced, in a dose-dependent manner to citrate. The percentages of neuroprotection by citrate (1, 2.5, and 5 mM) against $A{\beta}-induced$ toxicity were 19%, 31 %, and 34%, respectively. We report that citrate inhibits the formation of $A{\beta}$ fibrils in vitro and has neuroprotective effect against $A{\beta}$-induced toxicity in PC12 cells. Neuroprotective effects of citrate against $A{\beta}$ might be, to some extent, attributable to its inhibition of $A{\beta}$ fibril formation. Although the mechanism of anti-amyloidogenic activity is not clear, the possible mechanism is that citrate might have two effects, salting-in and surfactant effects. These results suggest that citrate could be of potential therapeutic value in Alzheimer's disease.

• Molecular & Cellular Toxicology
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• v.5 no.2
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• pp.126-132
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• 2009

The usage and types of chemicals are advancing, specializing, large-scaled increasing, and new chemical exposed workers are concerning to occupational disease. The generation of reactive oxygen in the body from carcinogen, mutation and DNA damage in cancer is protected by natural antioxidants (phytochemicals) with antimutagenic effect. There were many reports of ginsenoside Rb$_1$, Rg$_1$ grievances of the genetic mutation to suppress the effect confirm the genetic toxicity test with chromosomal aberration test and the Comet (SCGE) assay confirmed the suppression effect occurring chromosomal DNA damage. We had wanted to evaluate the compatibility and sensitivity between the chromosomal aberration (CA) test and the Comet assay. We used the CA test and Comet assay to evaluate the anti-genotoxicity of ginsenoside Rb$_1$ and Rg$_1$, in CHO-K1 (Chinese hamster ovary fibroblast) cell in vitro, composed negative control (solvent), positive control (benzo[a]pyrene), test group (carcinogen+variety concentration of ginsenoside) group. The positive control was benzo[a]pyrene (50 $\mu$M), well-known carcinogen, and the negative control was the 1 % DMSO solvent. The test group was a variety concentration of ginsenoside Rb$_1$, Rg$_1$ with 10$^{-8}$%, 10$^{-6}$%, 10$^{-4}$%, 10$^{-2}$%, 1%, 10%. In chromo-somal aberration test, we measured the number of cells with abnormally structured chromosome. In Comet assay, the Olive tail moment (OTM) and Tail length (TL) values were measured. The ratio of cell proliferation was increased 8.3% in 10$^{-8}$%, 10$^{-6}$%, 10$^{-4}$%, 10$^{-2}$%, 1%, 10% Rb$_1$ treated groups, and increased 10.4% in 10$^{-10}$%, 10$^{-8}$%, 10$^{-6}$%, 10$^{-4}$%, 10$^{-2}$%, 1% Rg$_1$ treated groups. In the CA test, the number of chromosomal aberration was decreased all the Rb$_1$ and Rg$_1$ treated groups. In the Comet assay, the OTM values were decreased in all the Rb$_1$ and Rg$_1$ treated groups. To evaluate the compatibility between CA and Comet assay, we compared the reducing ratio of chromosomal abnormalities with its OTM values, it was identified the antimutagenicity of ginsenoside, but it was more sensitive the CA test than the Comet assay. Ginsenoside Rb$_1$ and Rg$_1$ significantly decrease the number of cells with chromosomal aberration, and decrease the extent of DNA migration. Therefore, ginsenoside Rb$_1$, Rg$_1$ are thought as an antioxidant phytochemicals to protect mutagenicity. The in vitro Comet assay seems to be less sensitive than the in vitro chromosomal aberration test.

• Toxicological Research
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• v.14 no.3
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• pp.341-348
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• 1998

We established an in vitro experimental system using the following procedure. We first introduced tritium-labelled norepinephrine ([3H]-NE)into PC12 cells. The [3H]-NE incorporated into PC12 cells were then stimulated by a high concentration (60 mM) of $K^+$ during 12 minutes. Then, we counted the amount of [3H]-NE release from PC12 cells with the scintillation counter. After screening fungal, Streptomyces or bacterial product using this experimental system, we obtained S9940 from Streptomyces spp. which inhibited [3H]-NE release from PC12 cells. S9940 also inhibits the release of ATP as a neurotransmitter of PC12 cells and rat cortical neurons. The inhibitory effect was seen even when the PC12 cells were treated with low $K^+$ buffer containing ionomycin $(1\muM)$ as an ionopore. This result suggests that the inhibitory action of S9940 on neurotransmitter release appeared after the influx of $Ca^{2+}$.

• Toxicological Research
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• v.23 no.4
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• pp.331-339
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• 2007

Titanium or titanium alloy is a widely used implant material according to its certified biocompatibility, sufficient strength and ready availability. The purpose of this study was to evaluate the relative biocompatibility of titanium and titanium alloy specimens (Ti-29Nb-13Ta, TiNb and Ti-6Al-4V, Ti64) using in vivo and in vitro methods. For in vivo experiment, the specimens were implanted in the abdominal subcutaneous region of female mice for 2 and 4 weeks. The reaction of connective tissue to specimens was evaluated histologically. The specimens were encapsulated by fibrous connective tissue consisting of fibroblast, fibrocyte and other cells including neutrophil, macrophage, giant multinucleated cell and unidentified cells. Some newly formed blood vessels were located in the fibrous capsule surrounding the implant. Cell types and the thickness of fibrous capsules were examined quantitatively. Most of cell types located in the fibrous capsule were fibroblasts and fibrocytes. The average thickness of fibrous capsules for the TiNb specimens was much thinner than that of the titanium alloy, Ti64. The thickness of the fibrous capsule around all titanium specimens decreased at 4 weeks compared to 2 weeks post-implantation. The biocompatibility of titanium and titanium alloy specimens were also investigated in in vitro method using alkaline phosphatase from MG-63 cells. Alkaline phosphatase activity of the TiNb specimen showed higher activity than the titanium alloy, Ti64. In conclusion, the TiNb alloy with thin capsule thickness in vivo and high alkaline phosphatase activity in vitro will be of considerable use in biomedical applications.

• The Journal of Internal Korean Medicine
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• v.11 no.2
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• pp.22-42
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• 1990

Sabaiksan has been prescribed to treat various allergic diseases in herbal medicine which were induced by various vasoactive amine released from the mast cells. The constituents of Sabaiksan are Mori Cortex Radices(MCR), Lycii Cortex Radicis(LCR) and Glycyrrhizae Radix(GR). Recently, simple models of compound 48/80 induced anaphylactic shock and cutaneous reaction in vivo were developed to test various agents employed in the field of allergy and toxicology research. The purpose of this study is to evaluate the effects of Sabaiksan on compound 48/80 induced anaphylactic stock, cutaneous reaction and mesenteric mast cell degranulation rate in ICR mice, and on compound 48/80 induced peritoneal mast cell degranulation and histamine release in vitro. Groups of ICR mice were intraperitoneally pretreated with $100{\mu}{\ell}$ of saline, $MCR(2g/m{\ell}),\;LCR(2g/m{\ell}),\;GR(g/m{\ell})$ or Sabaiksan itself(MCR+LCR+GR) at 24, 12 and 1 hour before compound 48/80 solution ($10{\mu}{\ell}/gm$ B. W) were peritoneally given into them, and then mortality within 72 hours after the compound 48/80 injection, and mesenteric mast cell degranulation rate at 15 minutes after compound 48/80 injection were calculated. In vitro experiment, $400{\mu}{\ell}$ of rat peritoneal mast cell suspension$(10^6cell/m{\ell})$ were pretreated with $50{\mu}{\ell}$ of saline, $MCR(2g/m{\ell}),\;LCR(2g/m{\ell}),\;GR(g/m{\ell})$ or Sabaiksan itself at room temperature for 30 minutes, and then $50{\mu}{\ell}$ of compound 48/80 solution $(100{\mu}g/m{\ell})$ were added into it. 30 minutes after the addition of compound 48/80 solution, histamine release assay in the supernatant of peritoneal mast cell suspension were performed employing radioisotope enzymatic assay and morphologic changes of mast cells in each regular time point were photographed. Compared with controls, compound 48/80 induced anaphylactic shock was significantly inhibited by MCR and GR pretreatment into the ICR mice. Significant inhibition of compound 48/80 induced cutaneous reaction, mesenteric mast cell degranulation rate in vivo and histamine release from the rat peritoneal mast cells in vitro was observed only in MCR pretreated group. From the above results, it is suggested that MCR component of Sabaiksan may playa key role to suppress mast cell function since it has been applied to various allergic diseases.

• Journal of Ginseng Research
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• v.45 no.3
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• pp.401-407
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• 2021

Background: Gintonin is an exogenous ginseng-derived G-protein-coupled lysophosphatidic acid (LPA) receptor ligand. LPA induces in vitro morphological changes and migration through neuronal LPA1 receptor. Recently, we reported that systemic administration of gintonin increases blood-brain barrier (BBB) permeability via the paracellular pathway and its binding to brain neurons. However, little is known about the influences of gintonin on in vivo neuron morphology and migration in the brain. Materials and methods: We examined the effects of gintonin on in vitro migration and morphology using primary hippocampal neural precursor cells (hNPC) and in vivo effects of gintonin on adult brain neurons using real time microscopic analysis and immunohistochemical analysis to observe the morphological and locational changes induced by gintonin treatment. Results: We found that treating hNPCs with gintonin induced morphological changes with a cell rounding following cell aggregation and return to individual neurons with time relapses. However, the in vitro effects of gintonin on hNPCs were blocked by the LPA1/3 receptor antagonist, Ki16425, and Rho kinase inhibitor, Y27632. We also examined the in vivo effects of gintonin on the morphological changes and migration of neurons in adult mouse brains using anti-NeuN and -neurofilament H antibodies. We found that acute intravenous administration of gintonin induced morphological and migrational changes in brain neurons. Gintonin induced some migrations of neurons with shortened neurofilament H in the cortex. The in vivo effects of gintonin were also blocked by Ki16425. Conclusion: The present report raises the possibility that gintonin could enter the brain and exert its influences on the migration and morphology of adult mouse brain neurons and possibly explains the therapeutic effects of neurological diseases behind the gintonin administration.

• Journal of Ginseng Research
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• v.40 no.4
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• pp.325-333
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• 2016

Background: Ginseng extracts are known to have angiogenic effects. However, to date, only limited information is available on the molecular mechanism underlying the angiogenic effects and the main components of ginseng that exert these effects. Human umbilical-vein endothelial cells (HUVECs) are used as an in vitro model for screening therapeutic agents that promote angiogenesis and wound healing. We recently isolated gintonin, a novel ginseng-derived lysophosphatidic acid (LPA) receptor ligand, from ginseng. LPA plays a key role in angiogenesis and wound healing. Methods: In the present study, we investigated the in vitro effects of gintonin on proliferation, migration, and tube formation of HUVECs, which express endogenous LPA1/3 receptors. Results: Gintonin stimulated proliferation and migration of HUVECs. The LPA1/3 receptor antagonist, Ki16425, short interfering RNA against LPA1 or LPA3 receptor, and the Rho kinase inhibitor, Y-27632, significantly decreased the gintonin-induced proliferation, migration, and tube formation of HUVECs, which indicates the involvement of LPA receptors and Rho kinase activation. Further, gintonin increased the release of vascular endothelial growth factors from HUVECs. The cyclooxygenase-2 inhibitor NS-398, nuclear factor kappa B inhibitor BAY11-7085, and c-Jun N-terminal kinase inhibitor SP600125 blocked the gintonin-induced migration, which shows the involvement of cyclooxygenase-2, nuclear factor kappa B, and c-Jun N-terminal kinase signaling. Conclusion: The gintonin-mediated proliferation, migration, and vascular-endothelial-growth-factor release in HUVECs via LPA-receptor activation may be one of in vitro mechanisms underlying ginsenginduced angiogenic and wound-healing effects.