• Journal of Animal Science and Technology
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• v.55 no.4
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• pp.237-247
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• 2013

This study was to examine single or combined in vitro effects of environmental endocrine disruptors on boar sperm characteristics, oxidative stress damage in sperm and development of porcine IVF embryos. Addition of various concentration of NP (10, 20, $30{\mu}M$), DBP (10, 50, $100{\mu}M$) and BPA (1, 5 or $10{\mu}g/ml$) on boar sperm characteristics such as percentages of sperm motility, viability, membrane integrity and mitochondrial activity were dose-dependently decreased within 3, 6 or 9 hr incubation period (p<0.05). The overall detrimental effects increased with incubation time increasement. NP, DBP and BPA showed the detrimental effects on sperm membrane and mitochondria of energy production organelles affecting cell viability with the dependancy of dose and incubation time. In combination effects, NP ($10{\mu}M$) + DBP ($10{\mu}M$) significantly decreased boar general sperm characteristics for 3 or 6 hr incubation period compared with control (p<0.05). When both of NP and DBP concentrations (NP; $30{\mu}M$, DBP; $100{\mu}M$) increase, the detrimental effects on sperm characteristics were larger than those of low concentration combination (p<0.05). The inhibitory effects of NP ($30{\mu}M$) + BPA ($10{\mu}g/ml$) on sperm characteristics were larger than those of NP ($10{\mu}M$) + BPA ($1{\mu}g/ml$) (p<0.05). DBP ($100{\mu}M$) + BPA ($10{\mu}g/ml$) decreased sperm characteristics compared with the low concentration combination (DBP $10{\mu}M$ + BPA $1{\mu}g/ml$, p<0.05). This result indicates the detrimental effects of both chemicals on sperm characteristics were dose dependent. Addition of NP ($30{\mu}M$) + DBP ($100{\mu}M$), NP ($30{\mu}M$) + BPA ($10{\mu}g/ml$), DBP ($10{\mu}M$) + BPA ($1{\mu}g/ml$) or DBP ($100{\mu}M$) + BPA ($10{\mu}g/ml$) significantly increased lipid peroxidation for 3 or 6 hr incubation period (p<0.05) compared with no addition control. NP (${\geq}20{\mu}M$) decreased the percentages of IVF embryo development from morulae and blastocyst stages (p<0.05) and its detrimental effects were dose-dependant. BPA 0, 1, 5 or $10{\mu}g/ml$ decreased significantly and dose-dependently the percentage of morulae plus and blastocysts (p<0.05). Combinations of DBP ($100{\mu}M$) plus NP ($30{\mu}M$) and DBP ($100{\mu}M$) plus BPA ($10{\mu}g/ml$) did not affect on morulae and blastocyst development, but NP ($30{\mu}M$) plus BPA ($10{\mu}g/ml$) has significant detrimental effect on embryo development at these stages (p<0.05). These overall results indicate that the partial detrimental effects on boar sperm characteristics and embryo development by NP, DBP, BPA or the combination of these chemicals might be due to the increasement of lipid peroxidation and free radical formation in the cell and there were no specific interaction effects on boar sperm and embryo degeneration among the combined treatments.

• Asian-Australasian Journal of Animal Sciences
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• v.33 no.6
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• pp.879-887
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• 2020

Objective: Numerous studies have indicated that the apoptosis and proliferation of granulosa cells (GCs) are closely related to the normal growth and development of follicles and ovaries. Previous evidence has suggested that miR-126-3p might get involved in the apoptosis and proliferation of GCs, and phosphatidylinositol 3-kinase regulatory subunit 2 (PIK3R2) gene has been predicted as one target of miR-126-3p. However, the molecular regulation of miR-126-3p on PIK3R2 and the effects of PIK3R2 on porcine GCs apoptosis and proliferation remain virtually unexplored. Methods: In this study, using porcine GCs as a cellular model, luciferase report assay, mutation and deletion were applied to verify the targeting relationship between miR-126-3p and PIK3R2. Annexin-V/PI staining and 5-ethynyl-2'-deoxyuridine assay were applied to explore the effect of PIK3R2 on GCs apoptosis and proliferation, respectively. Real-time quantitative polymerase chain reaction and Western Blot were applied to explore the regulation of miR-126-3p on PIK3R2 expression. Results: We found that miR-126-3p targeted at PIK3R2 and inhibited its mRNA and protein expression. Knockdown of PIK3R2 significantly inhibited the apoptosis and promoted the proliferation of porcine GCs, and significantly down-regulated the mRNA expression of several key genes of PI3K pathway such as insulin-like growth factor 1 receptor (IGF1R), insulin receptor (INSR), pyruvate dehydrogenase kinase 1 (PDK1), and serine/threonine kinase 1 (AKT1). Conclusion: MiR-126-3p might target and inhibit the mRNA and protein expressions of PIK3R2, thereby inhibiting GC apoptosis and promoting GC proliferation by down-regulating several key genes of the PI3K pathway, IGF1R, INSR, PDK1, and AKT1. These findings would provide great insight into further exploring the molecular regulation of miR-126-3p and PIK3R2 on the functions of GCs during the folliculogenesis in female mammals.

• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.8
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• pp.919-928
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• 2017

The immuno-enhancing effects of alginate oligosaccharides from Sargassum coreanum were investigated. The alginate oligosaccharides were produced by an alginate-degrading enzyme from S. oneidensis PKA 1008. The degraded alginate oligosaccharides were visualized by thin-layer chromatography developed using a solvent system of 1-butanol/methanol/water, 4:1:2 (v/v/v). Alginate was degraded into dimmers at 60 h. As a result, the levels of Th1 cytokine [interferon $(IFN)-{\gamma}$ and interleukin (IL)-2] and Th2 cytokine (IL-6 and IL-10) increased with increasing incubation time compared to the control in vitro. Enzymatic extract treatment promoted proliferation of splenocytes at concentrations of 100 and 200 mg/kg at 24 h in vivo. Secretion of $IFN-{\gamma}$ and IL-2 significantly increased in a dose-dependent manner at 24 h as well as induced higher production of IgG2a in serum. Natural killer cell activity was measured and tended to increase. In addition, complete blood cell counts increased in a dose-dependent manner. These results indicate that alginate oligosaccharides produced by crude enzyme from S. oneidensis PKA 1008 may have significant immune activities.

• The Korean Journal of Food And Nutrition
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• v.28 no.3
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• pp.369-375
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• 2015

This study was performed to investigate the physiological activities of Ginkgo biloba sarcotesta extracts before and after heat treatment. G. biloba sarcotesta was heated at $130^{\circ}C$ for 2 h and extracted with water, 70% ethanol and 80% methanol. ABTS and DPPH radical scavenging activities increased after heating in the water (14.95 mg AAE/g and 7.36 mg TE/g) and ethanol extracts (12.20 mg AAE/g and 6.23 mg TE/g). ${\alpha}$-Glucosidase inhibitory activity decreased after heating in all but the water extract. Angiotensin converting enzyme I inhibitory activities decreased after heating in all extracts. Nitric oxide production inhibitory activity increased from 12.40~44.55% of the raw sample to 40.76~72.39% of the heated sample at a concentration of $200{\mu}g/mL$. Lipid accumulation inhibitory activities were similar before and after heat treatment. The highest antiproliferative effects on MCF-7 human breast cancer cell lines were observed in 80% methanol extract in the heated sample. Cell viability at concentrations of 25, 50, 100, and $200{\mu}g/mL$ measured 34.88, 17.58, 8.44 and 10.48%, respectively. From the results, the antioxidant and antiproliferative activities of G. biloba sarcotesta extracts increased with heat treatment, and research on the identification of the structure for the active compounds are needed in further studies.

• Restorative Dentistry and Endodontics
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• v.29 no.5
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• pp.470-478
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• 2004

The purpose of this study is to monitor the secretion of matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) produced by human osteosarcoma cell line (MG63) stimulated with Prevotella nigrescens lipopolysaccharides (LPS). and to compare the level of secretion before and after the treatment of calcium hydroxide on P. nigrescens LPS. LPS was extracted and purified from anaerobically cultured P. nigrescens. MG63 cells were stimulated by the LPS (0, 1, $10{\;}\mu\textrm{g}/ml$) or LPS($10{\;}\mu\textrm{g}/ml$) pretreated with 12.5 mg/ml of $Ca(OH)_2$ for 3 days. Total RNA was isolated from the cell. and real-time quantitative polymerase chain reaction (PCR) was performed for quantification of MMP-1 and TIMP-1. The results were as follows. 1. MMP-1 mRNA expression at 48 hr was highly increased by stimulation with P. nigrescens LPS. The increase was dose-dependent. 2. When stimulated with ($1{\;}\mu\textrm{g}/ml$ of LPS. TIMP-1 mRNA expression was highly increased at 24 hr and 48 hr. However. TIMP-1 expression was suppressed at higher concentration ($10{\;}\mu\textrm{g}/ml$). 3. When P. nigrescens LPS was pretreated with $Ca(OH)_2$. MMP-1 and TIMP-1 gene expression was downregulated. The results of this study suggest that transcriptional regulation of MMP-1 and TIMP-1 by P. nigrescens LPS could be one of the important mechanisms in bone resorption of periapical inflammation. The result of calcium hydroxide on MMP-1 and TIMP-1 gene expression suppression shows that calcium hydroxide detoxified bacterial LPS and thus should be used the medication of choice for intracanal dressings in root canal infected with black-pigmented bacteria.

• Environmental Analysis Health and Toxicology
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• v.18 no.2
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• pp.89-94
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• 2003

Tributyltin (TBT) used world-wide in antifouling paints for ships is a widespread environmental pollutant and cause reproductive organs atrophy in rodents. At low doses, antiproliferative modes of action have been shown to be involved, whereas at higher doses apoptosis seems to be the mechanism of toxicity in reproductive organs by TBT. In this study, we investigated that the mechanisms underlying DNA fragmentation induced by TBT in the rat leyding cell line, R2C. Effects of TBT on intracellular Ca$\^$2+/ level and reactive oxygen species (ROS) were investigated in R2C cells by fluorescence detector. TBT significantly induced intracellular Ca$\^$2+/ level in a time-dependent manner. The rise in intracellular Ca$\^$2+/ level was followed by a time-dependent generation of reactive oxygen species (ROS) at the cytosol level. Simultaneously, TBT induced the release of cytochrome c from the mitochondrial membrane into the cytosol. Furthermore, ROS production and the release of cytochrome c were reduced by BAPTA, an intracellular Ca$\^$2+/ chelator, indicating the important role of Ca$\^$2+/ in R2C during these early intracellular events. In addition, Z-DEVD FMK, a caspase-3 inhibitor, decreased apoptosis by TBT. Taken together, the present results indicated that the apoptotic pathway by TBT might start with an increase in intracellular Ca$\^$2+/ level, continues with release of ROS and cytochrome c from mitochondria, activation of caspases,and finally results in DNA fragmentation.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.42 no.1
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• pp.68-78
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• 1997

Laboratory and greenhouse studies were conducted to determine differential sensitivities on absorption of $^{14}$ C-oxyfluorfen and the anatomical responses in wheat and barley to protoporphyrinogen oxidase-inhibiting herbicides [oxyfluorfen (2-chloro-1- (3-ethoxy -nitrophen-oxy)-4-(trifluoromethyl) benzene, acifluorfen(5-[2-chloro-4-(trifluoromethyl) phenoxy]-2-nitro-benzoic acid), bifenox(methyl-5-(2, 4-dichlorophenoxy)-2-nitrobenzoate) and oxadiazon(5-tert-butyl-3-(2, 4-dichloro-5-isopropoxyphenyl)-1, 3, 4-oxadiazol-2-one)]. I$_{50}$ value of the tolerant wheat cultivars to oxyfluorfen was about 10$^{-4}$ , whereas that of the susceptible barley cultivars was about 10$^{-6}$ M, showing significant difference between the two groups. When foliage were applied with acifluorfen, bifenox or oxadiazon, the oxyfluorfen-tolerant wheat showed less decreased in shoot fresh weight and chlorophyll content than the susceptible barley. Also, when soil-applied with these herbicides test plants showed similar tendency in foliar application. Electrolyte leakage from the tissue treated with these compounds was the more influenced in the barley than the wheat. Malondialdehyde(MDA) production as index of lipid peroxidation was greater in the barley than the wheat by treatment of these compounds. Therefore, the differential sensitivities of wheat and barley to protoporphyrinogen oxidaseinhibiting herbicides was showed by our greenhouse and in vitro experiment. The absorption rates of $^{14}$ C-oxyfluorfen were higher in the barley than the wheat. And this tendency was showed appararitly difference by increase of treatment durations. After the oxfluorfen and oxadiazon treatment, the tolerant wheat did not show the structural damage in leaf surface, but the susceptible barley was damaged in the leaf waxy layer. However, the acifluorfen and bifenox treatment showed no difference between wheat and barley. The anatomical changes by these compounds treatment were not observed in the tolerant wheat but epidermal cell and mesophyll cell were highly broken in the susceptible barley.

• Journal of Life Science
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• v.18 no.2
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• pp.241-248
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• 2008

This study has been carried out to secretion antibodies for the purpose of preventing the infection of Helicobacter pylori and using them as a supplement for treatment. This experiments have been separated antigens from H. pylori and observed into antibody production and the agglutination of H. pylori for the separated antigens. As major antigenic proteins separated from H. pylori, the following could be verified: 12 kinds of band for whole cell (WC), seven kinds of band for outer membrane protein (OMP), three kinds of band for crude urease, and one kind of band for lipopolysaccharide (LPS). The IgG anti-H. pylori antibody of separated antigens showed $77.9{\pm}6.4{\mu}g/ml$ for we (L), $84.9{\pm}6.4{\mu}g/ml$ for OMP, and $123.8{\pm}2.9{\mu}g/ml$ for crude urease, at the same antigen concentration of $20{\mu}g/100ull$, which showed the most at the crude urease. And it turned out that the IgA antibodies were generated with $2.5{\pm}0.32{\mu}g/ml$ for WC (L), $2.0{\pm}0.43{\mu}g/ml$ for OMP, and $1.3{\pm}0.25{\mu}g/ml$ for crude urease, which demonstrated the most for WC (L) antigens. As a result of verifying the immunogenecity of antigenic protein through the Western blotting, major antigenic substances could be confirmed as follows: 10 kinds for WC, six kinds for OMP and three kinds for crude urease. The agglutination values on the H. pylori of the antibody were $2^5,\;2^5,\;2^6\;and\;2^7$ at the antigen serums of anti-WC (H), anti-WC (L), anti-OMP and anti-crude urease, respectively, which indicated the highest for the antigen serum of anti-crude urease. The urease activation-inhibiting absorbance of antigen serum created by each antigen was $0.14{\pm}0.01$ for WC (H), $0.16{\pm}0.01$ for WC (L), $0.18{\pm}0.03$ for OMP, and $0.18{\pm}0.04$ for urease, demonstrating a significant inhibiting effect, compared with $0.26{\pm}0.02$ of the control group.

• Journal of Korean Society of Forest Science
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• v.77 no.4
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• pp.382-388
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• 1988

Improvement of productivity by forest tree breeding work in Korea was estimated for a few important tree species. Progenies of 17 plus trees of red pine (Pines densiflora) outgrew by 57 percentage compared with progenies of unselected trees at age 15. If best three families are selected among the 17, more than double in volume grow-th is expected. The hybrid Pinus rigida ${\times}$ P. taeda showed more than double volume growth compare to P. rigida at a southern plantation at age 15. However, the superiority of the hybrid decreased at northern plantations, mainly because of low coldhardiness of the hybrid. At a northern plantation, the hybrid grew less than the P. rigida on upper hill, while the hybrid grew much better than the P. rigida on flat area. Another hybrid Populus alba ${\times}$ P. glandulosa grew faster than both parents by two to two and half times according to planting sites at age 10. Introduction of Pinus rigida also showed increased volume growth. Volume increase by selection of best five provenances among 45 at age 12 was estimated as 53 percent compare to progenies of plus trees in Korea, Additional 19 percent of volume increase was expected by selection of the best families within the best provenances. Annual production of chestnuts reached about 70,000 M/T by planting resistant clones to chestnut gall wasp (Dryocosmus kuriphilus), which killed almost all susceptible trees. Although polyploid trees and mutants have been produced by colchicine treatments in over 10 tree species, none of them is economically important Remarkable improvement of productivity is expected by biotechnology in future through selection, hybridization, introduction of foreign genes at cell, cell organelle and gene level, and gene transformation. At present, mass propagation of superior planting materials by tissue culture will increase the productivity.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.3
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• pp.487-496
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• 2013

Green coffee beans (CB, Indonesian Mandheling) were fermented with three kinds of mushrooms (Phellinus linteus, PL; Hericium erinaceum, HE; Ganoderma lucidum, GL) or two kinds of mycelia from molds (Monascus purpureus, MP; Monascus ruber, MR) using solid-state culture to enhance physiological activity. After the roasting of fermented green coffee beans, roasted coffees were extracted with a hot-water decoction or 95% ethanol reflux. Yields from hot water extracts (HW, 17.7~25.3%) were higher than those from ethanolic extracts (EE, 9.5~12.2%). Hot-water extracts of roasted coffees from green coffee beans fermented with two molds (MP-CB-HW and MR-CB-HW) showed higher total polyphenols, flavonoids, and DPPH free radical scavenging activity than roasted coffees from non-fermented (CB-HW) or fermented green coffee beans with the three mycelia from mushrooms. MR-CB-HW also had the most potent macrophage stimulating and mitogenic activity (1.32 and 1.40-fold of CB-HW, respectively). In addition, MP-CB-EE and MR-CB-EE did not show any cytotoxicity to the RAW 264.7 cell at a concentration of $100{\mu}g/mL$, and these extracts significantly inhibited nitric oxide (NO) production from the LPS-stimulated RAW 264.7 cell line (38.6 and 37.0% of the LPS-treated group). Meanwhile, the chlorogenic acid concentrations of MP-CB-HW or MR-CB-HW highly increased (to 76.21 or $76.73{\mu}g/mL$, respectively), but caffeine concentrations were not affected by solid-state fermentation. In conclusion, the physiological activities of roasted coffees were enhanced by the solid-state culture of green coffee beans with M. purpureus or M. ruber, suggesting that these roasted coffees could possibly serve industrial applications as functional coffee beverages.