Namhyun Chung

Cellular Toxicity of Various Inhalable Metal Nanoparticles on Human Alveolar Epithelial Cells

Nanoparticles (NPs) have a greater potential to travel through an organism via inhalation than any other larger particles, and could be more toxic due to their larger surface area and specific structural/chemical properties. The aim of this study was to evaluate in vitro biological effects of various inhalable metallic NPs (TiO2, Ag, Al, Zn, Ni). Human alveolar epithelial cells (A549) were exposed to various concentrations of NPs for 24 h. The extent of morphological damage was in the order of m-TiO2 > n-TiO2 > m-silica ≫ n-Ni ≈ n-Zn ≈ n-Ag ≈ n-Al and was affected in a dose-dependent manner. The extent of apoptotic damage measured with two-color flow cytometry was in the order of n-Zn > n- Ni > m-silica ≫ n- TiO2 > m- TiO2 > n-Al > n-Ag. The extent of apoptotic damage measured with DNA fragmentation was in the order of n-Zn ≈ m-silica > n- Ni ≫ m- TiO2 ≈ n- TiO2 ≈ n-Al > n-Ag, indicating no significant difference in the damages by both m-TiO2 and n-TiO2. The extents of apoptotic damages were also affected in a dose-dependent manner. Uptake of no other NPs but n-TiO2 and m-TiO2 into the cells was observed after 24 h exposure. The intracellular generation of ROS was significant with n-Zn but not with the other particles. These results demonstrated that various inhalable metallic NPs (TiO2, Ag, Al, Zn, Ni) could cause cell damages directly or indirectly. More detailed studies on the influence of size, structure, and composition of the NPs are needed to better understand their toxic mechanisms.

Effect of soil properties on bioavailability and extractability of phenanthrene and atrazine sequestered in soil

Sixteen soils with markedly different properties were analyzed to determine their porosity in the range of 7 nm-10 microm, cation-exchange capacity (CEC), surface area and clay mineralogy. The extent of sequestration of phenanthrene and atrazine has been shown to differ markedly among these soils. Correlations were sought between soil characteristics and four methods of measuring sequestration. Simple correlation analysis showed that some but not all measures of phenanthrene and atrazine sequestration were highly correlated with organic C content, nanoporosity or CEC but not other properties of the soils. Multiple linear-regression analysis suggested an interaction of organic C content with soil texture, CEC or surface area in determining the extent of atrazine or phenanthrene sequestration. We conclude that organic C content, CEC and other properties of soil may be useful predictors of sequestration of some compounds.

HOX gene analysis of endothelial cell differentiation in human bone marrow-derived mesenchymal stem cells

Human bone marrow-derived mesenchymal stem cells (hMSCs) have been shown to possess multilineage differentiation potential. HOX genes function in transcriptional regulators, and are involved in stem cell differentiation. The aim of the present study was to demonstrate HOX genes that are related to angiogenesis. To identify the expression patterns of 37 HOX genes in the endothelial cell differentiation of hMSCs, we analyzed HOX genes through profiling with multiplex RT-PCR. The results showed that the expression patterns of four HOX genes, HOXA7, HOXB3, HOXA3, and HOXB13, significantly changed during angiogenesis. The expression levels of HOXA7 and HOXB3 were dramatically increased, whereas those of HOXA3 and HOXB13 were decreased during endothelial cell differentiation. When further analysis of the expressions of these HOX genes was performed with real-time PCR and an immunoblot assay, the expression patterns were also found to be well-matched with the results of multiplex RT-PCR. Here, we report that HOXA7, HOXB3, HOXA3, and HOXB13 might be involved in the angiogenesis of hMSCs.

ROD-SHAPED IRON OXIDE NANOPARTICLES ARE MORE TOXIC THAN SPHERE-SHAPED NANOPARTICLES TO MURINE MACROPHAGE CELLS

Variable sizes of nanoparticles, ranging from nano to micro scale, are of toxicological interest. In the present study, the authors hypothesized that, in addition to the size, the shape of iron oxide (Fe2O3) nanoparticles is a major factor that contributes to particle cytotoxicity. Cytotoxicity to mouse macrophage cells (RAW 264.7) was investigated using 3 different particles: micro-sized Fe2 O3 (M-Fe2O3), nano-sized Fe2O3 (N-Fe2O3), and rod-shaped Fe2O3 (R-Fe2O3). Whereas M-Fe2O3 and N-Fe2O3 were located in the vacuole as aggregates, R-Fe2 O3 was often spread throughout the cytoplasm. The extent of cytotoxicity measured by the water soluble tetrazolium (WST-1) assay was in the order R-Fe2O3 ≈ N-Fe2O3 > M-Fe2O3, whereas the extent revealed by the lactate dehydrogenase assay was in the order R-Fe2O3 >> N-Fe2O3 ≈ M-Fe2 O3. In addition, the degree of tumor necrosis factor-α and reactive oxygen species (ROS) production was in the order of R-Fe2O3  > N-Fe2 O3 > M-Fe2O3. In addition, a much higher extent of necrosis was associated with the presence of R-Fe2O3. These results suggest that the higher degree of necrosis due to R-Fe2O3 is correlated with both the higher degree of membrane damage and ROS production by R-Fe2O3 compared with the results of the other Fe2O3 particles. These results also showed that the degree of cytotoxicity of nanoparticles should be evaluated based on shape as well as size, because changes in shape and size are accompanied by alterations in surface area, which relate closely to cytotoxicity.

Enhanced production of recombinant B-domain deleted factor VIII from Chinese hamster ovary cells by propionic and butyric acids

Sodium propionate, as well as sodium butyrate, enhanced the production of recombinant B-domain-deleted, factor VIII (rFVIIIdB) by Chinese hamster ovary cells growing in a spinner-flask with a protein-free medium by more than six-fold. The two acids, however, had different cytotoxicities.

Involvement of Oxidative Stress and Mitochondrial Apoptosis in the Pathogenesis of Pelvic Organ Prolapse

PURPOSE Biomechanical weakness of the pelvic supportive structures has been proposed to be a cause of pelvic organ prolapse. However, the molecular mechanism involved in these changes is not completely understood. In this investigation we evaluated oxidative stress biomarkers in the uterosacral ligaments of women with pelvic organ prolapse and compared them with those of women with normal support. In addition, mitochondrial apoptosis was examined. MATERIALS AND METHODS Samples were collected from 26 women with advanced stage pelvic organ prolapse and 29 age matched controls. The expression levels of 8-OHdG and 4-hydroxy-2-nonenal in the uterosacral ligaments were measured using immunohistochemistry. To assess mitochondrial apoptosis we performed TUNEL assay, immunohistochemistry for cleaved caspase-3 and cytochrome c, and Western blot analyses for cleaved caspase-3 and caspase-9. RESULTS The mean percentage of cells immunopositive for 8-OHdG, 4-hydroxy-2-nonenal, TUNEL, cleaved caspase-3 and cytochrome c in the uterosacral ligaments was significantly higher in patients with pelvic organ prolapse than in controls. Similarly, Western blot analysis revealed increased expression of cleaved caspase-3 and caspase-9 in patients with pelvic organ prolapse. Correlation analyses revealed significant positive correlations between the percentage of cells immunopositive for 8-OHdG or 4-hydroxy-2-nonenal and markers of mitochondrial apoptosis. Analyzing by pelvic organ prolapse quantification system stage according to C point, the mean percentage of cells immunopositive for 8-OHdG, 4-hydroxy-2-nonenal and cytochrome c was significantly higher in patients with pelvic organ prolapse compared to controls, regardless of stage. However, the mean percentage of TUNEL and cleaved caspase-3 positive cells was significantly higher only in patients with stage III or IV pelvic organ prolapse. CONCLUSIONS Oxidative stress and increased mitochondrial apoptosis may contribute to the pathological process of pelvic organ prolapse.

Berberine induces apoptosis via ROS generation in PANC-1 and MIA-PaCa2 pancreatic cell lines

Pancreatic cancer is the fourth leading cause of cancer death. Gemcitabine is widely used as a chemotherapeutic agent for the treatment of pancreatic cancer, but the prognosis is still poor. Berberine, an isoquinoline alkaloid extracted from a variety of natural herbs, possesses a variety of pharmacological properties including anticancer effects. In this study, we investigated the anticancer effects of berberine and compared its use with that of gemcitabine in the pancreatic cancer cell lines PANC-1 and MIA-PaCa2. Berberine inhibited cell growth in a dose-dependent manner by inducing cell cycle arrest and apoptosis. After berberine treatment, the G1 phase of PANC-1 cells increased by 10% compared to control cells, and the G1 phase of MIA-PaCa2 cells was increased by 2%. Whereas gemcitabine exerts antiproliferation effects through S-phase arrest, our results showed that berberine inhibited proliferation by inducing G1-phase arrest. Berberine-induced apoptosis of PANC-1 and MIA-PaCa2 cells increased by 7 and 2% compared to control cells, respectively. Notably, berberine had a greater apoptotic effect in PANC-1 cells than gemcitabine. Upon treatment of PANC-1 and MIA-PaCa2 with berberine at a half-maximal inhibitory concentration (IC50), apoptosis was induced by a mechanism that involved the production of reactive oxygen species (ROS) rather than caspase 3/7 activation. Our findings showed that berberine had anti-cancer effects and may be an effective drug for pancreatic cancer chemotherapy.

Development and evaluation of a protein microarray chip for diagnosis of hepatitis C virus.

A protein chip diagnostic kit was developed for the diagnosis of hepatitis C virus (HCV) based on the protein chip technique and the immuno-concentration method. This kit was designed for low-density protein chips and also for the availability of multiple sample screening. Applicability of the chip was evaluated using 96 blood specimens and the results were compared to results of an anti-HCV enzyme immunoassay (EIA) test. With further development, the technology associated with the development of this chip could be applied to the simultaneous detection of multiple protein-protein, protein-ligand interactions.

Contact and fumigant toxicities of 3-methylphenol isolated from ostericum koreanum and its derivatives against house dust mites

The acaricidal activities of an active constituent derived from Ostericum koreanum roots and its derivatives were determined using fumigant and direct-contact toxicity bioassays against Dermatophagoides farinae and Dermatophagoides pteronyssinus . This was compared with that of a commercial acaricide (benzyl benzoate). In the fumigant toxicity bioassay, 4-chloro-6-isopropyl-3-methylphenol (0.29 μg/cm(2)) was 37.17 times more effective than benzyl benzoate (10.78 μg/cm(2)) against D. farinae, followed by 6-fluoro-3-methylphenol (0.57 μg/cm(2)), 3-methylphenol (0.63 μg/cm(2)), 4-chloro-3-methylphenol (0.75 μg/cm(2)), and 4-isopropyl-3-methylphenol (0.78 μg/cm(2)). In the direct-contact toxicity bioassay, 4-chloro-6-isopropyl-3-methylphenol (0.21 μg/cm(2)) was 36.81 times more toxic than benzyl benzoate (7.73 μg/cm(2)) against D. farinae, followed by 6-fluoro-3-methylphenol (0.40 μg/cm(2)), 3-methylphenol (0.41 μg/cm(2)), 4-isopropyl-3-methylphenol (0.56 μg/cm(2)), and 4-chloro-3-methylphenol (0.60 μg/cm(2)). The acaricidal effects of 3-methyphenol derivatives against D. pteronyssinus were similar to those against D. farinae. In structure-activity relationships, acaricidal activities could be related to the introduction of chloro, fluoro, and isopropyl functional groups onto the 3-methylphenol skeleton. These results indicate that naturally occurring 3-methylphenol and its derivatives are potential house dust mite control agents.

Development of a varicella virus vaccine stabilizer containing no animal-derived component

Various stabilizers containing no animal-derived components were assessed for their efficacy in stabilizing live attenuated varicella virus under various storage temperatures. Stabilizers containing carrageenan, soy protein hydrolysates, or sucrose had a satisfactory performance to maintain infectivity of the cell-free virus when compared to control stabilizer containing animal-derived gelatin/gelatin hydrolysate.