• Korean Journal of Agricultural Science
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• v.46 no.3
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• pp.579-589
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• 2019

Due to the ban on the use of antibiotics, interest has been increasing for the development of therapeutic agents to treat various diseases using natural resources. Osthole, a natural coumarin compound used in traditional Chinese medicines, exerts an anti-inflammatory effect, but its effects in cows remain unknown. In this study, the effect of osthole on lipopolysaccharide (LPS)- or concanavalin-A (Con-A)- stimulated peripheral blood mononuclear cells (PBMCs) was assessed. Jugular venous blood was collected from Korean calves, and PBMCs were isolated. They were then used to study the immune response of PBMCs to treatment with osthole and LPS or Con-A for 72 h by measuring inflammatory cytokines including tumor necrosis factor-${\alpha}$ ($TNF-{\alpha}$) and interferon-${\gamma}$ ($IFN-{\gamma}$). Osthole significantly inhibited the mRNA secretion of $TNF-{\alpha}$ and $IFN-{\gamma}$ in a dose-dependent manner. Therefore, osthole inhibited LPS- or Con-A- induced $TNF-{\alpha}$ and Con-A-induced $IFN-{\gamma}$ production significantly in dose-dependent manner. These results clearly suggest that osthole inhibited the LPS- or Con-A- stimulated upregulation of pro-inflammatory cytokines in a dose-dependent manner, without causing obvious cytotoxic effects. Osthole could also protect cows from LPS- or Con-A- induced endotoxin shock, possibly by inhibiting the production of pro-inflammatory cytokines, which suggests that osthole might be a novel therapeutic agent for the prevention of inflammatory diseases.

• Korean Journal of Poultry Science
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• v.34 no.1
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• pp.31-36
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• 2007

This experiment was conducted to determine the effect of betaine on immune response in laying hens. A total of 72 ISA-brown laying hens were divided into four groups of 18 hens each and fed corn-soybean meal based diets with addition of 0, 300, 600 and 1,200 ppm betaine for four weeks. The effect of betaine on splenocyte proliferations with mitogens, concanavalin A(Con A) and pokeweed mitogen(PWM), were assayed after incubation using [3H] thymidine uptake. Proliferations of splenocyte were significantly increased by activation of mitogen Con A or PWM. Mitogen effects of Con A were increased by Con A plus betaine injection(0.1 mM), whereas PWM effects did not affect in PWM plus betaine injection(0.1 mM) in vitro. Splenocyte of laying hens fed betaine tended to proliferate in the presence of PWM, but appeared to be slightly suppressed in the presence of Con A in vivo. Proliferation of splenocytes which were stimulated by Con A or Con A+betaine injection(0.1 mM) were increased in dietary 600 ppm betaine, but inhibited in dietary 1,200 ppm betaine supplementation. Spleen weights and sheep red blood cell(SRBC) titers of hens fed betaine tended to increase compared to those of control, but were not significantly different. These results suggested that betaine could increase splenocyte proliferation in vitro.

• Parasites, Hosts and Diseases
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• v.27 no.2
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• pp.73-78
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• 1989

Cell-mediated and humoral immune reactions in mice infected with pathogenic Acanthamoeba culbertsoni were observed according to the period of time after amoebic infection by intranasal inoculation. The degrees of blastogenesis of spleen cells induced by mitogens, which were measured using radioactive [$^3H$]-thynndine, were compared between infected and non-infected control groups. The mitogens used in this blastogenesis experiment were concanavalin A (Con A) and lipopolysaccharide(LPS). On the other hand, enzyme-linked immunosorbent assay(ELISA) was employed for the detection of humoral antibodies against A, culbertsoni. The levels of blastogenesis of splenocytes and strum litres in the experimental group showed increasing tendency a week after inoculation of A. cuzberiseni, although there was no difference between the experimental and control groups in other periods of the experimental time.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.5
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• pp.713-719
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• 2006

The effect of dietary iodine and selenium supplementation, alone or in combination, on peripheral blood lymphocyte function was determined in laying hens. Eight-hundred-and-sixty-four New-Loman laying hens were randomly allotted into 12 dietary treatments with different inclusion levels of iodine (0, 0.1 and 0.2 mg/kg), selenium (0, 0.05, 0.1 and 0.2 mg/kg) or their combinations for 24 weeks. The lipopolysaccharide (LPS) stimulation index, concanavalin A (ConA) stimulation index, peroxide enzyme activity and phagocytosis to neutral red particles were tested. There were significant differences in LPS stimulation index, ConA stimulation index, peroxide enzyme activity and phagocytosis to neutral red particles in different iodine or selenium supplementation levels (p<0.05). The highest iodine and selenium supplementation both resulted in highest LPS-/ConA-stimulation indices (p<0.05). However, when iodine was lower than 0.2 mg/kg, the additional effect of different levels of selenium did not always result in significant differences in these indices. The results indicated that iodine and selenium may affect immunity in laying hens and, when the iodine level in the laying hen is lower than 0.2 mg/kg, a selenium allowance higher than 0.1 mg/kg may be necessary to improve immunity.

• Parasites, Hosts and Diseases
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• v.33 no.3
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• pp.195-200
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• 1995

The presence of biological response modifiers (BRMI-like effect was confirmed in peritoneal exudate (PE) of ToxopLnsmo gondii-infected ICR mice which inhibited Concanavalin A (Con A)-induced peritoneal Iymphocyte (PL) proliferation. During 5 days of PL incubation with $10{\;}\mu\textrm{g}/ml$ Con A with or without PE, 3H-thymidine uptake was measured for the last 24 hrs. Compared to uninduced control, PL proliferated by 7.3-fold with Con A induction_ When PE of infected mice was added, PL proliferation was inhibited by $74.0{\;}{\pm}{\;}11.9%$ whereas inhibition by PE of normal mice was $16.4{\;}{\pm}8.3%$. Inhibitory effect of PE increased exponentially from 3 days up to 4-5 days of survival after the infection. Inhibitory activity of PE was decreased concentration-dependently. Also the inhibition was diminished when the PE was treated with heat of $95^{\circ}C$ for 10 min orprecipitated with 10% trichloroacetic acid (TCA). In SDS-PAGE of PE, many minor bands appeared newly. Heat-labile protein molecule in PE exerted inhibitory activity to Con A- induced Iymphocyte proliferation.

• Molecules and Cells
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• v.38 no.11
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• pp.998-1006
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• 2015

Retinols are metabolized into retinoic acids by alcohol dehydrogenase (ADH) and retinaldehyde dehydrogenase (Raldh). However, their roles have yet to be clarified in hepatitis despite enriched retinols in hepatic stellate cells (HSCs). Therefore, we investigated the effects of retinols on Concanavalin A (Con A)-mediated hepatitis. Con A was injected into wild type (WT), Raldh1 knockout ($Raldh1^{-/-}$), $CCL2^{-/-}$ and $CCR2^{-/-}$ mice. For migration study of regulatory T cells (Tregs), we used in vivo and ex vivo adoptive transfer systems. Blockade of retinol metabolism in mice given 4-methylpyrazole, an inhibitor of ADH, and ablated Raldh1 gene manifested increased migration of Tregs, eventually protected against Con A-mediated hepatitis by decreasing interferon-${\gamma}$ in T cells. Moreover, interferon-${\gamma}$ treatment increased the expression of ADH3 and Raldh1, but it suppressed that of CCL2 and IL-6 in HSCs. However, the expression of CCL2 and IL-6 was inversely increased upon the pharmacologic or genetic ablation of ADH3 and Raldh1 in HSCs. Indeed, IL-6 treatment increased CCR2 expression of Tregs. In migration assay, ablated CCR2 in Tregs showed reduced migration to HSCs. In adoptive transfer of Tregs in vivo and ex vivo, Raldh1-deficient mice showed more increased migration of Tregs than WT mice. Furthermore, inhibited retinol metabolism increased survival rate (75%) compared with that of the controls (25%) in Con A-induced hepatitis. These results suggest that blockade of retinol metabolism protects against acute liver injury by increased Treg migration, and it may represent a novel therapeutic strategy to control T cell-mediated acute hepatitis.

• Journal of Life Science
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• v.17 no.5 s.85
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• pp.694-698
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• 2007

Berberine, an alkaloid initially isolated from chinese herbal medicine, has antibiotic activities against a variety of organisms including bacteria, viruses, fungi, protozoans, and chlamydia. Furthermore, berberine has shown a number of beneficial effects, including anti-tumor, anti-inflammation, and vasodilatory effects. In this work we have investigated the effects of berberine on lymphocyte proliferation and GM-CSF production in mice. Mouse splenocytes were incubated with berberine and concanavalin A(Con A) to observe the effects on cell proliferation. The culture supernatants of splenocytes exposed to berberine, berberine plus LPS, and berberine plus Con A were harvested to assay GM-CSF. The cell proliferation of nice splenocytes exposed to berberine only($1{\mu}g/ml$) was increased significantly more than PBS(control) group. But the Con A-induced cell growth was inhibited by berberine. The GM-CSF production from mice splenocyte culture exposed to berberine only was increased in comparison with PBS(control) group, but the production of it with LPS or Con A was inhibited by berbeline. The present findings may explain lympocyte proliferating and regulating effects of berberine.

• Journal of Sensor Science and Technology
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• v.27 no.4
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• pp.237-241
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• 2018

The real-time enrichment and detection of pathogens are serious issues and rapidly evolving field of research because of the ability of these pathogens to cause infectious diseases. In general, bacterial detection is accomplished by conventional colony counting or by polymerase chain reaction (PCR) after DNA extraction. As colony counting requires considerable time to cultivate, PCR is an attractive method for rapid detection. A small number of pathogens can cause diseases. Hence, a pretreatment process, such as enrichment is essential for detecting bacteria in an actual environment. Thus, in this study, we developed a microfluidic chip capable of performing rapid enrichment of bacteria and the extraction of their genes. A lectin, i.e., Concanavalin A (ConA), which shows binding affinity to the surface of most bacteria, was coated on the surface of magnetic particles to nonspecifically capture bacteria. It was subsequently concentrated through magnetic forces in a microfluidic channel. To lyse the captured bacteria, magnetic particles were irradiated by a wavelength of 532nm. The photo-thermal effect on the particles was sufficient for extracting DNA, which was consequently utilized for the identification of bacteria. Our device will help monitor the existence of bacteria in various environmental situations such as water, air, and soil.

• The Journal of the Korean Society for Microbiology
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• v.10 no.1
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• pp.1-8
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• 1975

Despite a number of recent studies on appendix its function appears to remain unknown. The present studies were undertaken in order to extend and confirm the previous studies concerning the role of appendix in immune response. An early hemagglutinin response of mercaptoethanol sensitive antibody(IgM antibody) in rabbit injected intravenously(i.v.) with 200mcg of bovine gamma globulin(BGG) was abolished by lethal whole body irradiation(900 r), but preserved in animals whose appendix and bone marrow were shielded during irradiation. Late formation of mercaptoethanol resistant antibody(IgG antibody) and the development of memory in bone marrow shielded animals were not affected by irradiation of the appendix. Formation of either IgM or IgG antibody to sheep red blood cells(SRBC) injected i.v. as determined by direct plaque forming cell(DPFC) technique in spleen were effectively abolished by appendectomy, thymectomy, or both followed by irradiation. When bone marrow was shielded in combination with autologous appendix reconstitution, DPFC response was about 5 times greater than the sum of two. Lysed appendix cells failed to restore the response. Lethally irradiated rabbits restored with combination of autologous appendix and thymus cells showed DPFC responses which were essentially normal. Three pools of appendix were obtained by manual separation technique and were stimulated with soluble concanavalin A(Con A), phytohemagglutinin-P(PHA) and pokeweed mitogen(PWM). Rabbit appendix cells responded to Con A, PHA and PWM. Cells of thymus dependent area(TDA) of the appendix were relatively enriched in their response to T cell mitogens compared to dome and follicle cells. The PHA/Con A responsive ratio of appenix TDA subpopulation was high, indicating that Con A responsive cells have a wider distribution among appendix. This finding showed that interfollicular area of the appendix is thymus-dependent. The present studies confirmed other evidence that the rabbit appendix cells itself are unable to form antibody and T lymphocytes in appendix TDA may be heterogenous, and that the appendix cells are synergistic with either bone marrow or thymus cells in the early hemagglutinin on splenic antibody response to BGG or SRBC.

• YAKHAK HOEJI
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• v.35 no.3
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• pp.154-164
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• 1991

These experiments were conducted to investigate the effects of Glycyrrhizae Radix extract(GR) on histamine synthesis, lymphocyte blastogenesis in C57BL/6J mice splenocytes, IL-1 production, $Ca^{2+}$ uptake by macrophage-like P388D$_{1}$ cells and plaque forming cell assay against SRBC. Histamine contents, lymphocyte blastogenesis, IL-1 activity, $Ca^{2+}$ uptake and plaque forming cell were determined by enzyme isotope method, [$^{3}$H]-thymidine incorporation, C3H/HeJ mouse thymocytes proliferation, the addition of 5 $\mu$Ci/ml $^{45}Ca^{2+}$ to P388D$_{1}$ cell suspension and assay to sheep red blood cell, respectively. Cytotoxicity, which was expressed as 50% mortality, was occurred by the addition of GR(10$^{-3}$g/ml). Histamine production in mouse spleen cell culture was significantly increased by 48 hour incubation added 0.25$\mu\textrm{g}$/ml of Con A. Con A-dependent T-lymphocyte proliferation was also enhanced by the addition of 0.25 $\mu\textrm{g}$/ml of Con A. GR depressed histamine contents at 10$^{-9}$~10$^{-4}$g/ml. and Con A (0.25 $\mu\textrm{g}$/ml) dependent T-lymphocyte proliferation at 10$^{-5}$~10$^{-4}$g/ml. IL-1 activity was significantly decreased by 10$^{-8}$~10$^{-4}$g/ml of GR. $Ca^{2+}$ uptake was not changed by GR, but antibody production markedly increased at 10.0~50.0 mg/kg of GR. From the above results, it is suggested that GR have immuno-regulatory action; GR decreased cell-mediated immune response and increased antibody production by B lymphocyte at high doses.