• 농업과학연구
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• 제46권3호
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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.

• 한국가금학회지
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• 제34권1호
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• pp.31-36
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• 2007

비태인이 산란계의 면역 반응에 어떠한 영향을 미치는지 알아보기 위하여 in vitro와 in vivo 시험을 시행하였다. 처리구당 18수씩 총 72수를 공시하였고, 비태인 0, 300, 600, 1,200 ppm 수준으로 산란계 사료에 첨가 급여하였다. 시험결과, in vitro 시험에서 비장 림프구 증식은 비태인 만을 처리하였을 때에는 증식 반응은 일어나지 않았으나 Con A와 PWM을 처리하였을 때 증식 반응은 현저하게 증가하였다. 또한 Con A만을 처리했을 때보다 Con A와 비태인을 동시에 처리했을 때 반응성이 현저하게 증가하였다. 그러나 PWM과 비태인을 동시에 처리했을 때는 PWM만을 처리했을 때와 비교해서 차이를 보이지 않았다. 사료에 비태인을 첨가 급여한 in vivo시험에서 Con A의 자극에 대한 비장 림프구 증식반응은 비태인의 첨가 수준에 따라 증가하는 경향을 보였으나 1,200 ppm 급여구에서는 대조구에 비하여 감소하는 경향을 보였다. 이러한 경향은 Con A와 함께 비태인(0.1 mM)을 처리했을 때도 동일하게 나타났다. PWM의 자극에 대한 반응은 Con A에 비하여 약하게 나타났으며 비태인 급여 수준에 따른 차이도 적게 나타났다. 비태인을 급여한 산란계 비장의 무게에서 처리구간에 통계적인 차이는 없었다. 비태인 첨가구는 대조구에 비하여 전체적으로 증가하는 경향을 보였고, 비태인 300 ppm 첨가구가 가장 높은 수치를 나타내었다. 비태인을 수준별로 급여하여 SRBC 항체가를 조사한 결과에서도 처리구간 유의적인 차이는 나타나지 않았다. 따라서 본 시험의 결과, 비태인의 처리는 in vivo 상에서 Con A로 비장 림프구의 증식을 유도한 경우 유의하게 비장 림프구의 증식을 증가시켰다.

• Parasites, Hosts and Diseases
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• 제27권2호
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• pp.73-78
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• 1989

병원성이 강한 Acanthamoeba culbertsoni를 마우스에 감염시킨 후 경과된 감염기간에 따른 세포매개성 면역반응과 혈청 내 항체가의 변동을 관찰하였다. [9H]-thymidine을 이용한 마우스 비장세포의 아세포화를 관찰하였는데 사용된 mitogen으로는 concanavalin A(Con A)와 lipopoITsaccharide(LPS)였으며, 항체의 측정을 위하여 효소표식 면역검사법(ELISA)을 시행하였다. T림프구 및 B림프구의 아세포화 정도는 A. culbertsoni 감염 7일 후 증가하는 경향을 보였으며 다른 기간에 서는 대조군에 비해 별다른 차이를 보이지 않았다. 또한 감염된 마우스의 혈청 내 항체가도 감염 7 일 후부터 증가하는 경향을 관찰할 수 있었다.

• Asian-Australasian Journal of Animal Sciences
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• 제19권5호
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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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• 제33권3호
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• pp.195-200
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• 1995

톡소포자충에 감염된 마우스의 복수액이 Con A로 유도한 정상 마우스 복수 림프구의 아세포화를 억제하는 효과를 관찰하였다. 마우스 감염은 톡소포자충의 RH주를 사용}하였다. ConA로 유도한 아세포화 정도는 정상 마우스의 복수 림프구를 얻어 96-well 배양기에 될주하고 Con A를 $10{\;}\mu\textrm{g}/ml$로 5일간 처리하면서, 4일째에 $^3H-thymidine$을 well 당 $1{\;}{\mu}Ci$씩 첨가하여 DNA 표지량으로 측정하였다. 아세포화 억제효과는 Con A와 감염 복수액을 동시 처리한 군의 아세포화를 Con A만 처리한 군에서의 아세포화에 대한 비율로 나타내었다. Con A에 의한 아세포화는 7.3배였다. 정상 마우스의 복수액에 의한 아세포화 억제효과는 $16.4{\;}{\pm}{\;}8.3%$이나, 감염 4일째 마우스의 복수액은 아세포화를 $74.0{\;}{\pm}{\;}11.9%$ 억제하였다. 생존일 4-5일의 감염 마우스에서 복수액을 일자별로 채취하여 처리하면 감염 1일에 $16.9{\;}{\pm}{\;}7.9%$, 2일에 $23.4{\;}{\pm}{\;}8.3%$, 3일에 $41.4{\;}{\pm}{\;}7.1%$ 및 4일에 $74.0{\;}{\pm}{\;}11.9%$로 높은 억제효과가 발현되었다. 감염후 4일에 얻은 복수액을 희석하여 처리하였을 때 아세포화 억제 효과는 농도에 따라 변화하였다. 감염복수액을 $95^{\circ}C$에서 10분간 열처리하면 억제 효과가 소멸되었으며, 10% TCA로 침전시킨 후 상층액으로 처리하여도 억제효과가 소멸하였다. 따라서 톡소포자충 감염 마우스 복수액에 복수 림프구의 아세포화를 억제하는 물질이 존재하며 그 생체활성물질은 열에 약한 단백질이라고 판단하였다.

• Molecules and Cells
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• 제38권11호
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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.

• 생명과학회지
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• 제17권5호
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• pp.694-698
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• 2007

본 연구에서는 berberine이 세포성 면역에 미치는 효과를 보기위하여 림프구증식과 GM-CSF생성을 보고자 하였다. 마우스 비장세포에 berberine을 작용시켰을 때 저농도에서 세포성장을 촉진함으로서 림프구증식에 자극제로서의 역할을 담당할 것으로 보인다. GM-CSF의 생성은 berberine 단독처리에 의해서 증가됨으로서 GM-CSF생성의 증가로 인한 세포성 면역반응의 상승이 예상되고 LPS나 Con A에 의한 GM-CSF의 생성을 저해함으로서 과도한 GM-CSF생성을 중간에서 차단할 것으로 보인다. 따라서 이러한 결과들은 앞으로의 추가적인 연구결과들이 있게 되면 임상에서의 면역요법과 항염증작용에 berberine이 이용될 가능성을 시사한다 하겠다.

• 센서학회지
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• 제27권4호
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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.

• 대한미생물학회지
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• 제10권1호
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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.

• 약학회지
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• 제35권3호
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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.