• 한국축산식품학회지
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• 제37권5호
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• pp.743-751
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• 2017

During slaughtering, animal blood is typically discarded, resulting in water pollution. However, this discarded blood has valuable components, such as immunoglobulin (Ig). Although several studies have been conducted to develop methods for effective recycling of slaughterhouse blood, they have not been commercially utilized in Korea. Here, we extracted an Ig-rich fraction from porcine blood that was then subjected to various in vitro tests, including pathogen growth inhibition, antigenic cross-reactivity, and anti-toxin activity. The porcine immunoglobulin concentrate (PIC) was effectively purified by eliminating other components, such as albumin, and consisted of approximately $63.2{\pm}2.9%$ IgG and $7.2{\pm}0.4%$ IgM on a protein basis. The results showed that it significantly suppressed the growth of pathogenic bacteria, and bound to all tested pathogens, including both gram-positive and gram-negative species, although the degree of activity differed according to strain. The PIC bound to two types of lipopolysaccharide (LPS) obtained from Escherichia coli O111:B4 and Salmonella enterica serotype typhimurium in a concentration-dependent manner. In addition, the PIC restored the proliferation activity of the lymphoblast K-562 cells when co-incubated with pathogenic LPS. These results confirm that the PIC prepared in this study is a potentially valuable functional food material or diet supplement as an alternative to antibiotics that can protect animals from pathogenic bacteria.

• 대한본초학회지
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• 제30권5호
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• pp.45-49
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• 2015

Objectives : The aim of this study is to investigate anti-inflammatory activity using various extracts of rice straw (DaoCao) extract (RS).Methods : To investigate the anti-inflammatory effect of RS, we examined the effect of RS on cytokines production on THP-1 cell. Cells were cultured in incubator (37℃, CO₂5%, 0.5% FBS-RPMI, 1X10⁶cells/ml). One hour after,Dermatophagoides pteronissinus(Dp., 10 ug/ml) was treated into cell and at 6 hour after, each different concentrations(0.1, 1 and 10 ug/ml) of RS were treated. The cells were incubated for 16 hours and harvest the supernatant. The levels of IL-4, IL-5, IL-6, IL-8, MCP-1 and TNF-αwere determined using a commercially available ELISA kit.Results : We investigated whether RS has the inhibition of inflammatory response in Jurkat cells and THP-1 cells. RS suppressed secretion of IL-4, IL-5, and TNF-αinduced by house dust mites in Jurkat cells. It showed significant effects for all concentrations. RS suppressed the increased expression of IL-6, IL-8 and MCP-1 after treatment with mite in THP-1 cells. These results suggest that RS may be used as a valuable agent for treating allergic diseases such as atopy due to its anti-inflammatory property.Conclusions : RS showed significant biological activities with anti-inflammatory in the human T cells. These results suggest that RS may be used as a valuable agent for treating allergic diseases such as atopy due to its anti-inflammatory property. In terms of Korean traditional medicine, we expect the results to contribute to building of EBM (Evidence-Based Medicine).

• The Korean Journal of Physiology and Pharmacology
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• 제1권6호
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• pp.657-664
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• 1997

The effects of adenosine, adenosine A1 receptor antagonist (DPCPX), or NMDA receptor antagonist (APV) on the spontaneous release of $[^3H]-5-hydroxytryptamine$ ($[^3H]-5-HT$) during normoxic/normoglycemic or hypoxic/hypoglycemic period were studied in the rat hippocampal slices. The hippocampus was obtained from the rat brain and sliced $400\;{\mu}m$ thickness with the tissue slicer. After 30 min's preincubation in the normal buffer, the slices were incubated for 30 min in a buffer containing $[^3H]-5-HT$ ($0.1\;{\mu}M,\;74{\mu}Ci/8\;ml$) for uptake, and washed. To measure the release of $[^3H]-5-HT$ into the buffer, the incubation medium was drained off and refilled every ten minutes through sequence of 14 tubes. Induction of glucose/oxygen deprivation (GOD; medium depleting glucose and gassed with 95% $N_2/5%\;CO_2$) was done in 6th and 7th tube. The radioactivities in each buffer and the tissue were counted using liquid scintillation counter and the results were expressed as a percentage of the total radioactivities. When slices were exposed to GOD for 20 mins, the spontaneous release of $[^3H]-5-HT$ was markedly increased and this increase of $[^3H]-5-HT$ release was blocked by adenosine ($10\;{\mu}M$) or DL-2-amino-5-phosphonovaleric acid (APV; $30\;{\mu}M$). Adenosine $A_1$ receptor specific antagonist, 8-cyclopentyl-1,3-dipropylxanthine (DPCPX) exacerbate GOD-induced increase of spontaneous release of $[^3H]-5-HT$. These results suggest that Adenosine may play a role in the GOD-induced spontaneous release of $[^3H]-5-HT$ through adenosine $A_1$ receptor activity.

• 한국식품영양과학회지
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• 제23권4호
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• pp.698-703
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• 1994

In spore rec-assay using B. subtillus H17(rec) and M 45(rec) , the ethanol extract of buckwheat leaves showed antimutagenicity in condition of low concentrations, but its did comutagenicity in condition of high concentrations. In Ames test, the ethanol extract of buckwheat leaves reduced the mtabenicity of N-methyl-N' -nitro-N-nitrosoguaidine (MNNG), benzo (a) pyrene(B($\alpha$)P), 2-amino-fluorene(2AF), and 3-amino -1, 4-dime-thyl-5-H-pyrido(4, 3-b) indol(Trp-P-1) in Salmonella typhimurium TA98 and TA100. The ethanol extract was fractionated by hexane, ethylacetate, butanol and water. Among Them hexane fraction showed the highest inhibition rate on the mutagenicity of B($\alpha$)P, and so did chloroform fraction on the mutagenicity of MNNG in S. typhimurium Ta98 and TA100. To elucidate the antimutagenic mechanism of the ethanol extract, it was mixed and co-incubated with various metagens, S9 mix, and the bacteria with different experimental orders and different reaction times. The ethanol extract did not affect reversion rate of pre-mutated. S.typhimurium. However, when the ethanol extract was added to the mutagens before their interaction with S.typhimurium , it reduced the mutation rate to 152$\pm$12-273$\pm$18 colonies/plates in case of MNNG, and 135$\pm$13-195$\pm$10 colonies/ plates in case of B($\alpha$)P), showing strong desmutagenic activity.

• 한국동물생명공학회지
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• 제34권3호
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• pp.240-246
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• 2019

The aim of this study was to investigate effect of heat stress on expression levels of plasminogen activators (PAs) related mRNAs and proteins, and changes of PAs activity in porcine endometrial explants. The endometrial explants (200 ± 50 mg) were isolated from middle part of uterine horn at follicular phase (Day 19-21) and were pre-incubated in serum-free culture medium at 38.5℃ in 5% CO₂ for 18 h. Then, the tissues were transferred into fresh medium and were cultured at different temperature (38.5, 39.5, 40.5 or 41.5℃) for 24 h. The expression level of urokinase-type PA (uPA), type-1 PA inhibitor (PAI-1), type-2 PAI (PAI-2), and heat shock protein-90 (HSP-90) mRNA were analysis by reverse-transcription PCR and proteins were measured by western blotting. The supernatant were used for measurement of PAs activity. In results, mRNA and protein levels of HSP-90 was higher in 41.5℃ treatment groups than other treatment groups (p < 0.05). The expression of uPA, PAI-1, and PAI-2 mRNA were slightly increased by heat stress, however, there were no significant difference. Heat stress condition suppressed expression of active uPA and PAI-2 proteins (p < 0.05), whereas PAI-1 protein was increased (p < 0.01). Although PAI-1 protein was increased and active uPA was decreased, PAs activity was greatly enhanced by exposure of heat stress (p < 0.05). These results suggest that heat stress condition could change intrauterine microenvironment through regulation of PAs activity and other factors regarding with activation of PAs might be regulate by heat stress. Therefore, more studies regarding with regulatory mechanism of PAs activation are needed.

• Journal of Animal Science and Technology
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• 제47권3호
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• pp.363-370
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• 2005

Experiments were conducted to determine the effects of beta-mercaptoethanol ($\beta$-ME) supplements to the in vitro maturation (IVM) medium on in vitro fertilization (IVF) and intracellular glutathione (GSH) concentration. Porcine cumulus-intact oocytes were matured in TCM-I99 medium containing porcine follicular fluid, sodium pyruvate, D-glucose, FBS, hormonal supplements, and $\beta$-ME (0, 25, 50 and 100 ${\mu}$M) for 36 to 46h. After culture, cumulus-free matured oocytes were co-incubated with epididymal spermatozoa for 18h. There were no significant differences in the maturation rate among treatment groups. However, increases (P < 0.05) in intracellular GSH concentration before and after. fertilization were observed in 50 ${\mu}$M $\beta$-ME supplements to the IVM medium. Also, increases (P < 0.05) in male pronuclear formation after IVF were observed in same treatment group. In conclusion, supplementing $\beta$-ME into the IVM medium increased intracellular GSH concentrations and increased fertilization in vitro.

• 한국수정란이식학회지
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• 제7권2호
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• pp.125-131
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• 1992

The nudear changes of bovine oocytes during 24 hrs. of culture for mejotic maturation were examined. Bovine oocytes were collected from small(<2 mm), medium(2~6 mm) and large(>6mm) follicles and classified into three grades by their morphological characteristics. A total of 242 oocytes collected were obtained:from 184 small, 157 medium and 1 large follicles, respectively and were classified into 95 grade I, 155 grade H and 92 grade III oocytes. All the bovine oocytes collected and graded were washed with a basal medium and incubated in groups of 10 for 24 hrs in 5% $CO_2$ and 39$^{\circ}C$. The basal medium used was composed of TCM-199 supplemented with sodium bicarbonate, sodium pyruvate, streptomycin, penicillin G and 10% FCS. The oocytes were cultured in drops of 50,$\mu$l basal medium supplemented with 35$\mu$g /ml FSH, 10$\mu$g /ml LH and 1$\mu$g /ml estradiol-17$\beta$. The oocytes were fixed and examined on their chromosomal status by 1% acetorcein staining in the interval of 3 hrs. Most of the grade I oocytes developed to germinal vesicule stage at 0 to 3 hrs., germinal vesicle breakdown at 6 hrs., metaphase I at 9 to 15 hrs., anaphase I and telophase I at 18 hrs., and metaphase II and the first polar body at 24 hrs. after culture for meiotic maturation. However, it was found that compared to grade I oocytes, grade H and W oocytes reached earlier to germinal vesicle breakdown and most of them developed earlier to M II stage at 21 hrs. after culture.

• 한국식품영양학회지
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• 제28권5호
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• pp.759-765
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• 2015

The effects of extracts from Lentinula edodes (L. edodes) and Aquilariae agallocha (A. agallocha) on the DNA damage response in ultraviolet A (UVA)-exposed HaCaT cells and on the allergic contact dermatitis caused by 2,4-dinitro-chlorobezene (DNCB) were investigated. When UVA-exposed cells were incubated for 24 hours in medium containing L. edodes or A. agallocha extract, the level of 8-OHdG and CPD decreased in a concentration-dependent manner. The combined treatment with both extracts potentiated the decrease in UVA-induced 8-OHdG and CPD levels as compared with those following treatment with a single extract. In addition, the two extracts showed preventive effects against the UVA-induced reduction in collagen levels. Furthermore, the blood levels of IgE, IL-6, and histamine decreased more significantly upon combined treatment with L. edodes and A. agallocha extracts as compared with those following treatment with single extracts in DNCB-induced allergic contact dermatitis in the ICR mouse. The results of the present study suggest that the components with in the extracts of L. edodes and A. agallocha can help to prevent of UVA-induced genomic instability via a decrease in DNA damage, and to decrease the DNCB-induced allergic dermatitis via modulation of relevant proteins including IgE and IL-6. Further study is needed to clarify the purified components related to the preventative effects of the two extracts against UVA- or DNCB-induced genomic damage.

• 대한약침학회지
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• 제15권4호
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• pp.42-51
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• 2012

Objectives: This study used the basic principle of Oriental medicine, the sovereign, minister, assistant and courier principle (君臣佐使論) to investigate the effects of the component of ONGABO, which is composed of Ginseng Radix (Red Ginseng), Angelica Gigantis Radix, Schisandrae Fructus, Cuscuta Semen and Curcumae tuber on the viability of HepG2 cells. Methods: Single and mixed extracts of the component of ONGABO were prepared by lypohilizing powder of Red Ginseng (6-year root from Kanghwa), Angelica Gigantis Radix, Schisandrae Fructus, Cuscuta Semen, Curcumae Tuber (from Omniherb Co., Ltd., Korea) at the laboratory of herbal medicine in Woosuk University and were eluted after being macerated with 100% ethanol for three days. The cell viability of HepG2 was determined by using an absorptiometric analysis with PrestoBlue (Invitrogen) reagent after the plate had been incubated for 48 hours. All of the experiments were repeated three times to obtain the average value and standard deviation. The statistical analysis was done and the correlation factor was obtained by using Microsoft Office Excel 2007 and Origin 6.0 software. Results: Although Ginseng Radix (Red Ginseng) and Schisandrae Fructus did not enhance the viability of HepG2 cells, they were shown to provide protection of those cells. On the other hand, Angelica Gigantis Radix decreased the viability of HepG2 cells significantly, Cuscuta Semen and Curcumae Tuber had a small or no effect on the viability of HepG2 cells. Conclusions: In the sovereign, minister, assistant and courier principle (君臣佐使論), Ginseng Radix (Red Ginseng) corresponds to the sovereign component because it provides cell protection effects, Angelica Gigantis Radix corresponds to minister medicinal because it kills cells, Schisandrae Fructus corresponds to the assistant medicinal to help red ginseng having cell protect effects. Cuscuta Semen and Curcumae Tuber correspond to the courier medicinal having no effect in cell viability in HepG2. We hope this study provides motivation for advanced research on the sovereign, minister, assistant and courier principle.

• 동의생리병리학회지
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• 제25권5호
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• pp.857-862
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• 2011

The purpose of this study is to investigate the anti-proliferative mechanism by Trichosanthes kirilowii (TCK) in MCF-7 human breast carcinoma cell. In this study, we used human breast cancer cell line, Michigan cancer foundation-7 cells (MCF-7 cells). They were co-incubated with 30~200 ${\mu}g$/ml TCK for 48 hours, and cell viability was measured by Water-soluble tetrazolium salt-1 (WST-1) assay. After MCF-7 cells were exposed to 60 ${\mu}g$/ml of TCK for 0, 3, 6, 12, 24, 48 hours, We performed flow analysis cytometry sorting(FACS) and western blot analysis. We investigated the effect of dose-dependent cell growth inhibition by TCK, which could be proved by WST-1 assay. Also, flow cytometry analysis showed that TCK increased percentage of subG1 phase and G2/M phase cell cycle. In addition, TCK induced apoptosis through the expression of caspase-9, -3 and poly(ADP-ribose) polymerase(PARP) activation. Moreover, we showed that ATM-dependent G2/M phase arrest by DNA damage and phosphorylation of chk2, cdc25C, cdc2(Tyr15). Taken together, these results suggest that by G2/M phase arrest through DNA damage and inducing of apoptosis through intrinsic pathway, TCK may have potential tumor suppressor in breast cancer.