• Asian-Australasian Journal of Animal Sciences
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• v.22 no.1
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• pp.35-41
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• 2009

This study was undertaken to evaluate how ${\beta}$-mercaptoethanol (bME), an exogenous antioxidant, interacts with preantral follicles cultured in vitro. Mouse primary or secondary follicles were cultured in glutathione (GSH)-free or GSH-containing medium supplemented with bME of various concentrations, and the growth of preantral follicles, the maturation of intrafollicular oocytes and preimplantation development after parthenogenesis were monitored. In experiment 1, 0, 25, 50 or 100 ${\mu}M$ bME was added to culture medium supplemented with 100 ${\mu}M$ GSH or not. When secondary follicles were cultured in GSH-free medium, no significant change in follicle growth was detected after bME addition. However, exposure to bME in the presence of GSH significantly inhibited both follicle growth and oocyte maturation. Such detrimental effect became prominent in primary follicles and bME strongly inhibited follicle growth in the absence of GSH. In conclusion, there are stage-dependent effects of bME on follicle growth and oocyte maturation, and selective use of antioxidants contributes to establishing an efficient follicle culture system.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.129.2-130
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• 2003

Tomato soil pathogens(Phythopthora spp.) analyed high rates in series culture soil and existed in culture parts. To make a diagnosis of Phythopthora sp. and Its concentration, potato slices were manufactured to a round shape(2.5cm) or retangular form(1x4cm). and then, The potato slices dipped into diagnostic reagents with an antibiotic substance for 2∼4hours. Potato slices treated with a few reagents varied into 15cm depths in innoculated soils for 24hrs. Mycelium of the Phytophthora root rot fungus, Phythopthora capsici, were produced easily on potato slice. We collected many potato slice samples on diseased fields in various area. After storage of 24hrs in 20$^{\circ}C$ incubator, White mycelium of Phythopthora sp. formed on potato slice surface. Dilute concentrations of Phythopthora sp. was detected very low contents(1${\times}$10$^1$sporangia/g). But expressing Phythopthora root rots on potato slice did not developed larger lesions upon storage time in room temperature. These results suggest that the use of potato slice in a series of soil cultural system may still serve as efficient means of diagnosis of Phythopthora root rots in the absence of control measures.

• Journal of Veterinary Clinics
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• v.21 no.3
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• pp.236-242
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• 2004

1,2-benzopyrone can stimulate macrophages to increase the ability of phagocytosis. Peripheral blood polymorphonuclear cells (PMN) and macrophages destroy microbial organisms with reactive oxygen species (ROS), called oxidative burst activity (OBA). This study was undertaken to determine whether 1,2-benzopyrone affects the OBA on the phagocytic response of canine peripheral blood phagocytes. The OBA of phagocytes in the addition or absence of latex beads was analyzed by flow cytometry system using dihydrorhodamine 123 (DHR). The direct treatments of 1,2-benzopyrone have no effect on the OBA of peripheral blood mononuclear cells (PBMC), PMN and monocyte-rich cells regardless of addition of latex beads. When latex beads are added to PMN, the OBA of PMN was remarkably enhanced by culture supernatant from PBMC but not PMN treated with 1,2-benzopyrone. Similary, it was also enhanced by human recombinant (hr) $TNF-\alpha.$ However, when latex beads were not added to PMN, its OBA was not enhanced by culture supernatant from either PBMC or PMN treated with 1,2-benzopyrone. The OBA of latex beads-phagocytized PBMC and monocyte-rich cells was not enhanced by culture supernatant from either PBMC or PMN treated with 1,2-benzopyrone. These results strongly suggested that 1,2-benzopyrone has an immunoenhancing effect on the OBA of PMN when phagocytic response occurred only. This enhanced OBA may be mediated through active humoral substance(s), such as $TNF-\alpha,$ produced by PBMC stimulated with 1,2-benzopyrone.

• Journal of Environmental Health Sciences
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• v.38 no.1
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• pp.1-7
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• 2012

Carbon nanotubes (CNTs) stand at the frontier of nanotechnology and are destined to stimulate the next industrial revolution. Rapid increase in their production and use in the technology industry have led to concerns over the effects of CNT on human health and the environment. The prominent use of CNTs in biomedical applications also increases the possibility of human exposure, while properties such as their high aspect ratio (fiber-like shape) and large surface area raise safety concerns for human health if exposure does occur. It is crucial to develop viable alternatives to in vivo tests in order to evaluate the toxicity of engineered CNTs and develop validated experimental models capable of identifying CNTs' toxic effects and predicting their level of toxicity in the human respiratory system. Human lung epithelial cells serve as a barrier at the interface between the surrounding air and lung tissues in response to exogenous particles such as air-pollutants, including CNTs. Monolayer culture of the key individual cell types has provided abundant fundamental information on the response of these cells to external perturbations. However, such systems are limited by the absence of cell-cell interactions and their dynamic nature, which are both present in vivo. In this review, we suggested two viable alternatives to in vivo tests to evaluate the health risk of human exposure to CNTs.

• Biomolecules & Therapeutics
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• v.18 no.1
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• pp.16-23
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• 2010

Adipocyte differentiation in human bone marrow mesenchymal stem cells (hBM-MSCs) is not as efficient as that in murine pre-adipocytes when induced by adipogenic agents including insulin, dexamethasone, and 3-isobutyl-1-methylxanthine (IDX condition). Therefore, the promotion of adipocyte differentiation in hBM-MSCs has been used as a cell culture model to evaluate insulin sensitivity for anti-diabetic drugs. In hBM-MSCs, $PPAR{\gamma}$ agonists or sulfonylurea anti-diabetic drugs have been added to IDX conditions to promote adipocyte differentiation. Here we show that troglitazone, a peroxisome proliferator-activated receptor-gamma ($PPAR{\gamma}$) agonist, significantly reduced the levels of anti-adipogenic $PGE_2$ in IDX-conditioned hBM-MSC culture supernatants when compared to $PGE_2$ levels in the absence of $PPAR{\gamma}$ agonist. However, there was no difference in the mRNA levels of cyclooxygenases (COXs) and the activities of COXs and prostaglandin synthases during adipocyte differentiation in hBM-MSCs with or without troglitazone. In hBM-MSCs, troglitazone significantly increased the mRNA level of 15-hydroxyprostaglandin dehydrogenase (HPGD) which can act to decrease $PGE_2$ levels in culture. These results suggest that the role of $PPAR{\gamma}$ activation in promoting adipocyte differentiation in hBM-MSCs is to reduce anti-adipogenic $PGE_2$ levels through the up-regulation of HPGD expression.

• Reproductive and Developmental Biology
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• v.30 no.4
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• pp.301-305
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• 2006

The aim of this study was to investigate whether addition of porcine epididymal fluid (pEF) into culture medium during in vitro maturation influences the nuclear maturation of porcine germinal vesicle (GV) oocytes. Porcine cumulus-oocyte complexes (COCs) from follicles were cultured in tissue culture medium 199 (TCM 199) containing pEF. After 48hr of culture, oocytes were examined for evidence of GV breakdown, metaphase I, anaphase-telophase I, and metaphase II. The proportion of oocytes reaching at metaphase II (M II) stage was significantly (p<0.05) increased in oocytes cultured in the media supplemented with 10% pEF during in vitro maturation than in those without pEF regardless of cumulus presence or absence (54.6% vs 22.5%, 51.7% vs 24.2%). The supplementation of pEF during maturation of oocyte enhanced oocytes maturation in a dose-dependent manner in vitro. Also significant differences (p<0.05) in the percentage of MII oocytes were observed according to exposure period in pEF. Present study suggests that pEF contains a enhancing component(s) for nuclear maturation of porcine immature oocytes in vitro.

• Journal of Life Science
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• v.8 no.4
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• pp.387-394
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• 1998

A thermophilic bacterium (strain DG0303) producing a thermostable $\alpha$-glucosidase was isolated from manure and identified as Bacillus sp. Strain DG0303 produced high level of $\alpha$-glucosidase compared with other thermophilic Bacillus strains. The cellular protein patterns were also compared with other Bacillus strains by sodium dodecyl sulfatepolyacrylamide gel electrophoresis(SDS-PAGE). On the basis of 16S rDNA analysis the Bacillus sp. DG0303 was found to be a member of Bacillus rDNA group 5. The optimum temperature for growth was 65$\circ$C and no growth was obtained at 40$\circ$C or 75$\circ$C. The optimum pH for growth was 5.5 to 8.5. $\alpha$-glucosidase activity was produced during growth and most activity was detected in the culture supernatant. The $\alpha$-glucosidase production was constitutive in the absence of carbohydrates. High level of enzyme activity was detected when the culture was grown on medium containing starch. Addition of glucose resulted in the repression of the $\alpha$-glucosidase production. The optimum pH and tempoerature for enzyme activity were pH 5.0 and 65$\circ$C, respectively. When analyzed by zymogram, the culture supernatant showed a single $\alpha$-glucosidase band with a molecular weight of approximately 60,000.

• Microbiology and Biotechnology Letters
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• v.22 no.4
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• pp.360-367
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• 1994

The effect of various nutrients for the production of the anti-complementary polysaccha- ride by Flammulina velutipes, was examined and the partial purification of the polysaccharide was carried out. The culture conditions and medium compositions considerably influenced the anti-complementary activity of the polysaccharides. When a culture was carried out at 23$\circ$C and pH 5.5 for 6 days in the synthetic medium supplemented with galactose and NaNO$_{3}$, the production of the anti-complementary polysaccharide was maximized. The crude polysaccharide, FV-1 was isolated from the culture broth and partially purified into two fractions, FV-1-II and FV-1-III by gel filtration using Sephadex G-100. FV-1-II was mainly consisted of xylose, mannose, galactose and glucose, and rhamnose, mannose and galactose, for FV-1-III. Also, the anti-complementary activity of FV-1-III was reduced partially in the absence of the Ca ion. When crossed immunoelect- rophoresis using anti-human C3 serum was carried out after incubation of normal human serum with the FV-1-III in the Ca ion free condition, a cleavage of C3 precipitin line was observed. These results indicate that the mode of complement activation by polysaccharide from Flammulina velutipes is via not only the classical pathway but also the alternative pathway.

• Journal of Microbiology and Biotechnology
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• v.9 no.6
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• pp.726-731
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• 1999

It was discovered that ammonium phosphate in the medium played an important role in both growing mycelium and producing exopolysaccharides (EPS) from G. lucidum. In lower concentration levels of ammonium phosphate (0-3 g/l), an improved mycelial growth was observed by maintaining more filamentous morphology than in high concentrations (5-11 g/l). In addition, it was confirmed by comparing the factual dimension and frequency of the area regarding the mycelial pellets. This must be attributed to limitations of nutrient transfer by maintaining filamentous mycelium during the cultivation in a low ammonium phosphate containing medium. On the other hand, the best EPS production was observed in medium with the absence or low concentration of ammonium phosphate. The shear stress of the culture broth was greatly affected by the shear rate, as compared with that of the culture broth with high ammonium phosphate concentration. The rheological characteristics of the fermentation broth and filtrate worked well according to the Herschel-Bulkley model. It was also found that the morphological changes of the mycelium resulting from the ammonium phosphate concentration directly affected the rheological characteristics of the system and resulted in reversely affecting the EPS production levels. Based on these results, it can be concluded that delicate regulation of the ammonium phosphate concentration in the culture media should be provided in order to obtain optimal mycelial growth and/or EPS production.

• Journal of Microbiology and Biotechnology
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• v.22 no.2
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• pp.239-243
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• 2012

Degradation of butylbenzyl phthalate (BBP) by the white rot fungus Pleurotus ostreatus and the activities of some degrading enzymes were examined in two different media containing 100 mg/l of the compound. P. ostreatus pre-grown for 7 days in complex YMG medium was able to completely degrade BBP within an additional 24 h but degraded only 35 mg/l of BBP in 5 days of incubation in minimal medium. Fungal cell mass in the culture in YMG medium was higher in the presence than in the absence of BBP. The esterase activity of the fungal culture in YMG medium was higher than that in minimal medium and increased with the addition of BBP. On the contrary, laccase activity was higher in minimal medium and it did not increase upon the addition of BBP. General peroxidase activity increased for a few days after the addition of BBP to both media. The degradation of BBP and its metabolites by P. ostreatus thus may be attributed mostly to esterase rather than lignin-degrading laccase. In addition, the activities of the enzymes involved in BBP degradation and their changes varied significantly in the different media and culture conditions.