• Molecular & Cellular Toxicology
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• v.1 no.2
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• pp.124-129
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• 2005

Pro-inflammatory cytokines, such as tumor necrosis factor $(TNF)-{\alpha}$, interleukin-12 (IL-12) and interleukin $(IL-1)-{\beta}$, play a key role in causing inflammatory diseases, which are rheumatoid arthritis, Crohn's disease and sepsis. Accumulating evidences suggest that low level laser irradiation (LLLI) may have an anti-inflammatory action. However, there are few data regarding down regulation of Th1 immune response by using the diod typed laser emitting device for human patients. As a fundamental step in order to address this issue, we investigated immunological impact of the low level laser irradiation (10 mw laser diode with a wavelength of 630 nm) on expression of pro-inflammatory cytokines in murine immunocytes (splenocytes and peritoneal macrophages) in vitro. The LLLI on lipopolysaccharide (LPS 100 ng/ml)-stimulated murine splenocytes and macrophages, clearly down regulated mRNA expression of $TNF-{\alpha}$ and IL-12 in dose-dependent manner. In addition, LLLI significantly inhibits the NO production in the LPS-stimulated murine macrophages. This data suggests that LLLI (wavelength of 630 nm) may exert an anti-inflammatory action via modulation of pro-inflammatory cytokine and NO production pathway.

• Molecular & Cellular Toxicology
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• v.1 no.2
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• pp.106-110
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• 2005

N-ethyl-N-nitrosourea (ENU), which is a toxin and a carcinogen, as well as a mutagen, has a variety of effects on mice. ENU induces point mutation in male germ cell. Number of mutant animals are developed with ENU treatment. However, potentiality ot ENU as a carcinogen is not fully understood, even though, mutagenicity of ENU is broadly studied, In the present study, the gene expression profiling and histopathological investigation of ENU treated mouse's liver and brain were investigated. Also, the expression patterns of cancer related genes in ENU-treated mouse were analyzed.

• Molecular & Cellular Toxicology
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• v.1 no.2
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• pp.78-86
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• 2005

Dioxins, especially 2, 3, 7, 8-Tetrachlorodibenzo-p-dioxin (TCDD or dioxin), are ubiquitous environmental contaminants. TCDD is known that it has toxic effects in animals and humans, including chloracne, immune, reproductive and developmental toxicities, carcinogenicity, wasting syndrome and death. TCDD induces a broad spectrum of biological responses, including disruption of normal hormone signaling pathways, reproductive and developmental defects, immunotoxicity, liver damage, wasting syndrome and cancer. Many researches showed that TCDD induces gene expression of transcriptional factors related cell proliferation, signal transduction, immune system and cell cycle arrest at molecular and cellular levels. These toxic actions of TCDD are usually mediated with AhR (receptor, resulted from cell culture, animal and clinical studies). cDNA microarray can be used as a highly sensitive and informative marker for toxicity. Additionally, microarray analysis of dioxin-toxicity is able to provide an opportunity for the development of candidate bridging biomarkers of dioxin-toxicity. Through microarray technology, it is possible to understand the therapeutic effects of agonists within the context of toxic effects, classify new chemicals as to their complete effects on biological systems, and identify environmental factors that may influence safety.

• Molecular & Cellular Toxicology
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• v.1 no.2
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• pp.87-91
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• 2005

Although 2D-PAGE has been widely used as the primary method for protein separation, difficulties in displaying proteins with an extreme values of isoelectric paint (pI), molecular size and hydrophobicity limit the technique. In addition, time consuming steps involving protein transfer and extraction from the gel-pieces can result in sample loss. Here, we describe a novel protein separation technique with capillary size-exclusion chromatography (CSEC) for rapid protein identification from human plasma. The method includes protein fractionation along with molecular size followed by in-solution tryptic digestion and peptide analysis through reversed phase liquid chromatography (RPLC) coupled to nanoflow electrospray-tandem mass spectrometry (ESI-MS/MS). Tryptic peptides are applied an a $100\;{\mu}m\;i.d.{\times}10mm$ length pre-column and then separated on a $75\;{\mu}m{\times}200mm$ analytical column at -100 nL/min flaw rate. Proteins were identified over the wide ranges of pI (3.7-12.3) when this technique was applied to the analysis of $1-2\;{\mu}L$ of human plasma. This gel-free system provides fast fractionation and may be considered a complementary technique to SDS-PAGE in proteomics.

• Molecular & Cellular Toxicology
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• v.5 no.2
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• pp.108-112
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• 2009

Alzheimer's disease(AD) is a neurodegenerative disorder characterized pathologically by senile plaques, neurofibrillary tangles, and synapse loss. Especially, extracellular beta-amyloid (Abeta) deposition is a major pathological hallmark of Alzheimer's disease (AD). In AD senile plaques, high level of iron and car-bonylated Abeta were detected. Iron has a Lewis acid property which can increase the electrophilicity of carbonyls, which may react catalytically with nucleophiles, such as amines. Hence, this study investigated whether or not iron could promote the carbonylation of amine with malondialdehyde (MDA) in the physiological condition. As the basic study, we examined that iron might promote the conjugation reaction between propylamine, monoamine molecule and MDA in the physiological condition. As the concentration of iron increased, the fluorescence intensity produced from the conjugation reaction increased in a dose-dependent manner. Instead of propylamine, we applied the same reaction condition to Abeta 1-40 peptide, one of major components founded in AD senile plaques for the conjugation reaction. As the result, the fluorescence intensity produced from the conjugation reaction between Abeta 1-40 peptide and MDA showed the similar trend to that of the reaction used with propylamine. This study suggests that iron can accelerate the conjugation reaction of MDA to Abeta 1-40 peptide and play an another important role in deterioration of AD brain.

• Molecular & Cellular Toxicology
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• v.5 no.1
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• pp.14-22
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• 2009

In vitro cytotoxicity of 23 alkyl phenols (APs) on human cervical cancer cell lines (HeLa) was determined using the lactate dehydrogenase (LDH) cytotoxicity assay. Two different sets of descriptors were used to construct the calibration model based on Genetic Algorithm-Multiple Linear Regression (GA-MLR) based on the experimental data. A statistically robust Structure-Activity Relationships (QSAR) model was achieved ($R^2$=95.05%, $Q^2_{LOO}$=91.23%, F=72.02 and SE= 0.046) using three Dragon descriptors based on Me (0D-Constitutional descriptor), BELp8 (2D-Burden eigenvalue descriptor) and HATS8p (3D-GETAWAY descriptor). However, external validation could not fully prove its validity of the selected QSAR in characterization of the cytotoxicity of APs towards HeLa cells. Nevertheless, the cytotoxicity profiles showed a finding that 4-n-octylphenol (4-NOP), 4-tert-octyl-phenol (4-TOP), 4-n-nonylphenol (4-NNP) had a more potent cytotoxic effect than other APs tested, inferring that increased length and molecular bulkiness of the substituent had important influence on the LDH cytotoxicity.

• Molecular & Cellular Toxicology
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• v.5 no.1
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• pp.23-31
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• 2009

To compare the effects of nanometer-sized silver ions and support materials (nano-silver coating material, NM-silver) and silver ions, we exposed zebrafish embryos to both types of nano-silver ions and compared the acute responses during embryogenesis. The amount of silver in the NM-silver (17.16%) was greater than that in the silver ion (4.56%). Both of these materials have different atomic compositions. The silver ion-exposed groups (10 and 20 ppt) showed lower survival rates than the NM-silver-exposed groups (10 and 20 ppt). NM-silver penetrated the skin and blood tube of zebrafish larvae as aggregated particles, whereas, silver ions penetrated the organelles, nucleus and yolk in a spread-out pattern. Micro-array analysis of RNA from zebrafish larvae (72 hours post-fertilization) that were treated with either NM-silver or silver ions, showed alteration in expression of the BMP, activin, TGF-$\beta$, and $GSK3{\beta}$ genes pathway. Additionally, $GSK3{\beta}$ gene pathway for apoptosis that was related with left-right asymmetry. Gene expression changes in the NM-silver or silver ions-treated zebrafish embryo led to phenotypic changes in the hatched larvae, reflecting increased apoptosis and incomplete formation of an axis.

• Molecular & Cellular Toxicology
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• v.5 no.1
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• pp.7-13
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• 2009

In this study, we investigated the anti-proliferative effect of Damina 909 in human cancer cell lines and tumor-xenografted nude mice to elucidate its potential in treating many cancers. Damina 909 treatment resulted in inhibition of cell proliferation of human pancreatic cancer cells. Our in vivo study showed that the weight of pancreatic tumors in Damina 909-treated group were the lighter than control group. Consequently, the intake of food and water in Damina 909-treated group did not change, while those in control group were steadily decreased over a period of treatment. Moreover, Damina 909 treatment elevated the protein expression of p53 and p21 in pancreatic tumor of xenografted nude mice. In summary, compare to other human cancer cells such as prostate and hepatocyte, Damina 909 is most effectively inhibited proliferation of pancreatic cancer cells by increasing the expression of tumor suppressor genes. This led us to speculate that a candidate substance for effective cancer therapy of pancreatic cancer might be contained in Damina 909.

• Molecular & Cellular Toxicology
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• v.5 no.1
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• pp.1-6
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• 2009

Microglial cells constitute the first line of defense against infection and injury in the brain. This study was conducted to evaluate the protective mechanisms of Agrimonia pilosa (AP) on LPS-induced activation of BV-2 microglial cells. The effects of AP on gene expression profiles in activated BV-2 microglial cells were evaluated using microarray analysis. BV-2 microglial cells were cultured in a 100 mm dish ($1{\times}10^7/mL$) for 24 hr and then pretreated with 1 g/mL AP or left untreated for 30 min. Next, 1 g/mL LPS was added to the samples and the cells were reincubated at $37^{\circ}C$ for 30 min, 3 hr and 6 hr. The gene expression profiles of the BV-2 microglial cells varied depending on the AP. The microarray analysis revealed that MAPK signaling pathway-related genes were down-regulated and IL10 gene was up-regulated in AP-treated BV-2 microglial cells. AP can affect the inflammatory response and MAPK pathway in BV-2 microglial cells.

• Molecular & Cellular Toxicology
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• v.5 no.1
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• pp.44-50
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• 2009

This study was conducted to evaluate the inflammation mechanisms of tumor necrosis factor-$\alpha$ (TNF-$\alpha$), interleukin-4 (IL-4), and IL-$1{\beta}$-induced stimulation of A549 human epithelial cells. In the present study, A549 cells were stimulated with TNF-$\alpha$, IL-4 and IL-$1{\beta}$ to induce expression of chemokines and adhesion molecules involved in eosinophil chemotaxis. The effects of TNF-$\alpha$, IL-4 and IL-$1{\beta}$ on gene expression profiles in A549 cells were evaluated by oligonucleotide microarray and Real time RT-PCR. The gene expression profiles for the A549 cells varied depending on the cytokines. Also, the results of the microarray and Real time RT-PCR revealed that inflammatory-related genes were up-regulated in cytokine stimulated A549 cells. Cytokines can affect inflammation in A549 cells. A microarray-based genomic survey is a high-throughput approach that enables evaluation of gene expression in cytokine stimulated cell lines.