• Ocean and Polar Research
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• v.34 no.2
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• pp.137-149
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• 2012

Hemocyte parameters of the wild Pacific oyster Crassostrea gigas inhabiting intertidal zones in small bays (Gwangyang and Jinhae Bay) on the southern coast of Korea were evaluated using flow cytometry and neutral red retention (NRR) assay. Morphological features, cell count, mortality, DNA damage, phagocytosis, and lysosomal membrane stability of hemocytes were analyzed. Three types of hemocytes were identified in the oyster hemolymph: granulocytes, hyalinocytes, and blast-like cells. Immune related functions of hemocyte including phagocytosis and lysosomal membrane stability were significantly different among the study areas (P<0.05), while cell count, mortality, and DNA damage of hemocytes were not significantly different. In Gwangyang Bay, phagocytosis of granulocytes and lysosomal membrane stability of oyster hemocytes inhabiting inside bay were significantly lower than those of oyster hemocytes in outside bay (P<0.05), indicating that oysters in inside bay of Gwangyang were relatively suppressed the immunological function in hemocytes. Contrary to Gwangyang Bay, immune parameters of oyster hemocytes in Jinhae Bay not showed the difference between sampling sites. In conclusion, flow cytometry and NRR assay using oyster hemocyte has a powerful tool to investigate the cell level in a short time due to no-preprocessing of material.

• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.8
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• pp.1057-1065
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• 2012

The effects of volatile flavor extracts of eight different herbal medicines, Juniperus rigida (JR), Saussurea lappa SL), Cnidium officinale (CO), Angelica gigas (AG), Eugenia caryophyllata (EC), Angelica tenuissima (AT), Mentha arvense (MA), and Artemisiae argyi (AA), were investigated on LPS-stimulated inflammation using Raw 264.7 cells. The volatile flavor extracts of CO and AG considerably inhibited LPS-stimulated NO, $PGE_2$, IL-6, and TNF-${\alpha}$ (except AG) production, as well as iNOS expression. Major volatile components of CO were identified as ligustilide and of ${\beta}$-eudesmol as AG by GC-MS analysis. Thus, these results suggest that the volatile extracts of CO and AG may be useful as potential therapeutic agents for inflammation-associated disorders.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.3
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• pp.334-339
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• 2007

Reactive oxygen species (ROS) generate during electrical activation of oocytes which has detrimental effects on embryo survival when overwhelmed. The present study was designed to investigate the ability of L-ascorbic acid, a novel water soluble antioxidant, to reduce the ROS level in developing embryos and their subsequent effects on embryo development in vitro. The compact cumulus oocyte complexes (COCs) were cultured in tissue culture medium (TCM)-199 supplemented with 10 ng/ml epidermal growth factor, 4 IU/ml pregnant mare serum gonadotropin (PMSG), and human chorionic gonadotropin (hCG) and 10% (v/v) porcine follicular fluid (pFF) for 44 h. After maturation culture, the denuded oocytes were activated with a single DC pulse of 2.0 kV/cm in 0.3 M mannitol solution containing 0.5 mM of HEPES, 0.1 mM of $CaCl_2$ and 0.1 mM of $MgCl_2$ for $30{\mu}s$ using a BTX Electro-cell Manipulator. The activated oocytes were cultured in modified North Carolina State University-23 (mNSCU-23) medium for 168 h. The level of $H_2O_2$ in each embryo was measured by the dichlorohydrofluorescein diacetate (DCHFDA) method at 48 h after activation. The blastocyst formation rate was significantly higher when culture medium was supplemented with 50 and $100{\mu}M$ L-ascorbic acid (31.2 and 38.7%, respectively) compared to non-supplemented (16.1%) group. Accordingly, significantly more cells in blastocyst were found for 50 and $100{\mu}M$ L-ascorbic acid (50.0 and 56.4, respectively) compared to 0 and $200{\mu}M$ L-ascorbic acid (36.5 and 39.8, respectively). L-ascorbic acid reduces the $H_2O_2$ level in developing embryos in a dose-dependant manner. The $H_2O_2$ level (pixels/ embryos) was 191.5, 141.0, 124.0 and 163.3 for 0, 50, 100 and $200{\mu}M$ L-ascorbic acid, respectively. So, we recommend to supplement 50 or $100{\mu}M$ L-ascorbic acid in porcine in vitro culture medium.

• IMMUNE NETWORK
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• v.10 no.6
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• pp.239-246
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• 2010

Background: Monoclonal antibodies (mAbs) recognizing Class III epitope of CD34 are essential for flow cytometric diagnosis of leukemia. Methods: 27H2 mAb was developed from a mouse alternatively immunized with human acute leukemia cell lines, KG1 and Molm-1. Using flow cytometric analysis of various leukemic cell lines and peripheral blood, immunohistochemical study of frozen tonsil, we characterized 27H2 mAb. Antigen immunoprecipitated with 27H2 mAb immunobloted with anti-CD34 mAb. A case of bone marrow sample of acute lymphoblastic leukemia (ALL) patient was obtained at CBNU Hospital. For epitope identification enzyme treatment with neuraminidase and O-sialoglycoprotein endopeptidase (OSGE) and blocking assay with known classIII mAb (HPCA-2) were done. Results: Only KG1 and Molm-1 revealed positive immunoreactivity. Immunohistochemical staining disclosed strong membranous immunoreactivity on high endothelial venules. Antigen immunoprecipitated by 27H2 mAb showed approximately 100 kDa sized band immunoblotted with anti-CD34 under non-reducing conditions. Epitope recognized by 27H2 mAb disclosed resistancy to both neuraminidase and OSGE treatment and completely blocked with known class III mAb preincubation. CD34 positive leukemic cells in BM of pre B cell ALL patient detected by FITC-conjugated 27H2 and HPCA-2 were identified with similar sensitivity. Conclusion: A novel murine mAb recognizing class III epitope of human CD34 with high affinity, which is useful for flow cytometric diagnosis of leukemia, was developed.

• Journal of Nutrition and Health
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• v.38 no.4
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• pp.307-312
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• 2005

We investigated the growth inhibitory effects of Atrina pecitinata (AP) on the proliferation in human cancer cell lines in vitro. AP was extracted with methanol which was further fractionated into four different types: methanol (APMM), haxane (APMH), butanol (APMB), and aquous layers (APMA). Among various partition layers, the APMM showed the strongest cytotoxic effects on all cancer cell lines which we used. In the MTT assay of AP fractions, the growth inhibitory effects was increased in proportion to its concentration. We observed quinone reductase (QR) induced effects in all fraction layers of AP on HepG2 cells. The QR induced effects of APMM on HepG2 cell at 80 $\mu$g/mL concentration indicated 2.0 with a control value of 1.0.

• Applied Microscopy
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• v.40 no.4
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• pp.201-209
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• 2010

Korean traditional herbs, especially Anemarrhena asphodeloides (A. asphodeloides), Salvia miltiorrhiza (S. miltiorrhiza), and Terminalia chebula (T. chebula) have been known to have the immunomodulatory, anti-tumor, and anti-inflammatory effects. However, direct cellular mechanism underlying the mast cell-mediated anaphylaxis-like reaction has poorly been understood. In the present study, the effects of the methanol extracts of A. asphodeloides (MEAA), S. miltiorrhiza (MESM), and T. chebula (METC) on anaphylactic reaction were investigated. Using in vitro and in vivo experiments, the influences of MEAA, MESM, and METC on the compound 48/80-induced anaphylaxis-like reaction and mast cell activation, and IgEmediated PCA were examined. Results are below; 1) MEAA, MESM, and METC significantly inhibited systemic anaphylaxis-like reaction, ear swelling response, and IgE-mediated PCA. 2) the compound 48/80-induced mast cell degranulation, histamine release of rat peritoneal mast cells (RPMC) were significantly inhibited by the pretreatment with MEAA, MESM, and METC, and 3) the compound 48/80-induced calcium influx in RPMC was significantly inhibited by the pretreatment with MEAA, MESM, and METC. These results suggest that MEAA, MESM, and METC may be an activity to inhibit the compound 48/80-induced or anti-DNP IgE-mediated anaphylactic reactions by blocking histamine release and calcium uptake into RPMC. MEAA, MESM, and METC potentially may serve as an effective therapeutic tool for allergic diseases.

• Journal of Plant Biotechnology
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• v.29 no.4
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• pp.229-233
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• 2002

Ferritin is ubiquitous in bacteria, animals and plants. Ferritin is thought to play two main roles in living cells to provide iron for the synthesis of iron protein such as ferretoxin and cytochromes and to prevent damage from radicals produced by iron/dioxygen interaction. To enhance the resistance of potato to Erwinia carotovora, the soybean ferritin gene was introduced into the potato either under CaMV 35S or hsr203J promoter. Potato transgenic plants were screened by PCR analysis using specific primers to the ferritin gene. Expression of ferritin gene under CaMV 35S and hsr203J promoter in potato transgenic plants was confirmed by northern blot analysis. hsr203J promoter known to pathogen inducible in tobacco drives the induction upon Phytophthora infestan in potato and the transcript level of ferritin gene was extremely high after 24 hours post inoculation. One of transformants under CaMV 35S promoter was increased 2.5 fold than untransformant. Each one of transgenic potato containing gene promoter CaMV 35S and hsr203J-ferrtin fusion exhibited tolerance against potato soft rot.

• Korean Journal of Veterinary Research
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• v.34 no.1
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• pp.25-36
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• 1994

The inward tail current after a short depolarizing pulse has been known as Na-Ca exchange current activated by intracellular calcium which forms late plateau of the action potential in rabbit atrial myocytes. Chloride conductance which is also dependent upon calcium concentration has been reported as a possible tail current in many other excitable tissues. Thus, in order to investigate the exsitance of the calcium activated chloride current and its contribution to tail current, whole cell voltage clamp measurement has been made in single atrial cells of the rabbit. The current was recorded during repolarization following a brief 2 ms depolarizing pulse to +40mV from a holding potential of -70mV. When voltage-sensitive transient outward current was blocked by 2 mM 4-aminopyridine or replacement potassium with cesium, the tail current were abolished by ryanodine$(1{\mu}M)$ or diltiazem$(10{\mu}M)$ and turned out to be calcium dependent. The magnitudes of the tail currents were increased when intracellular chloride concentration was increased to 131 mM from 21 mM. The current was decreased by extracellular sodium reduction when intracellular chloride concentration was low(21 mM), but it was little affected by extracellular sodium reduction when intracellual chloride concentration was high(131 mM). The current-voltage relationship of the difference current before and after extracellular sodium reduction, shows an exponential voltage dependence with the largest magnitude of the current occurring at negative potentials, with is similar to current-voltage relationship at negative potentials, which is similar to current-voltage relationship of Na-Ca exchange current. The current was also decreased by $10{\mu}M$ niflumic acid and 1 mM bumetanide, which is well known anion channel blockers. The reversal potentials shifted according to changes in chloride concentration. The current-voltage relationships of the niflumic acid-sensitive currents in high and low concentration of chloride were well fitted to those predicted as chloride current. From the above results, it is concluded that calcium activated chloride component exists in the tail current with Na-Ca exchange current and it shows the reversal of tail current. Therefore it is thought that in the physiologic condition it leads to rapid end of action potential which inhibits calcium influx and it contributes to maintain the low intracellular calcium concentration with Na-Ca exchange mechanism.

• Applied Chemistry for Engineering
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• v.16 no.4
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• pp.491-495
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• 2005

Vanadium pentoxide xerogels with a doping ratio of $Al/V_2O_5$ ranging from 0.01 to 0.05 were synthesized by doping Al into $V_2O_5$ xerogel via the sol-gel process. By using the synthesized $Al_xV_2O_5$, the $Li/Al_xV_2O_5$ cells were assembled to investigate the chemical and electrochemical properties. Surface morphology of the $Al_xV_2O_5$ xerogel showed an anisotropic corrugated sheet-like matrix, and the interlayer distance was about $11.5{\AA}$. The IR spectra of the $Al_xV_2O_5$ revealed that the doped Al was coordinated to the vanadyl group in $V_2O_5$. The $Al_xV_2O_5$ xerogels showed enhanced reversibility and energy density compared with the $V_2O_5$ xerogel. The specific capacity of the $Al_{0.05}V_2O_5$ xerogel was more than 200 mAh/g at 10 mA/g discharge rate, and cycle efficiency was about 90% after the 31st cycling test between 1.9 V and 3.9 V.

• Journal of Food Hygiene and Safety
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• v.33 no.5
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• pp.389-398
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• 2018

The purpose of this study was to evaluate anti-obesity activity of Cirsium setidens Nakai test material (CNTM) in 3T3-L1 adipocytes and obese C57BL/6J mice fed with a high-fat diet using various obesity-related in vitro experiments. During adipocyte differentiation, CNTM significantly inhibited lipid accumulation and ROS production compared to controls. To evaluate whether CNTM could exert glycerol release effects on mature 3T3-L1 adipocytes, we treated cells with various concentrations of CNTM for 1 h. Treatment of mature adipocytes with $160-320{\mu}g/mL$ of CNTM increased the release of glycerol, but not in a significant dose-dependent manner. Anti-adipogenic and anti-lipogenic effects of CNTM seemed to be mediated by the inhibition of $PPAR{\gamma}$ and $C/EBP{\alpha}$. Moreover, CNTM stimulated fatty acid oxidation in an AMPK-dependent manner. CNTM-treated groups of C57BL/6J mice showed reduced body weights and adipose tissue weight with improving serum lipid profiles and adiponectin protein expression in obese C57BL/6J mice fed with a high-fat diet. These results suggest that CNTM might have anti-obesity effect on adipogenesis and lipid metabolism in vitro and in vivo. This presents the possibility of developing a treatment for obesity using nontoxic natural resources.