• 한국식품영양과학회지
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• 제23권6호
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• pp.1032-1037
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• 1994

A simple, reproducible , and accurate enzymatic method using a cholesterol assay kit was developed to quantify total cholesterol content in egg yolk. Total egg yolk lipid was extracted with hexane : isopropanol(3 : 2, v/v) mixture. Samples containing various amount of the total lipid(0-3mg) in optically identifical culture tubes were reacted for 10 min in a water bath (37$^{\circ}C$) with the enzyme solution (5ml) from the cholesterol assay kit. Cholesterol content of the reaction mixturesin culture tubes was spectrophotometrically determined by two different ways : (1) using the culture tube as a curvette(designate culture tube method ; CTM) and (2) the quartz cvette containing the reaction mixture transferred from the culture tube (designate standard cvette method, SCM). CTM revealed lower cholesterol content in 0.1-1.0mg lipid sample range that SCM did, but not significant. For more than 2.0mg lipid sample, CTM gave significantly (p<0.01) lower cholesterol content relative to that by SCM, suggesting that SCM give a false positive result from the sample containing more than 2 mg lipid due to the interference of absorbance by lipid dispersed in the reaction solution . Cholesterol content of less than 1.0mg lipid sample by CTM was proportional to the amount of lipid used, but its linear relationship was not seen in more than 2mg lipid sample. Thus, to determine the appropriate lipid amounts (mg) analyzed . A constant level (41$\mu\textrm{g}$/mg) of cholesterol concentration was observed from the sample containing 0.1-1mg lipid. after which the cholesterol level was dropped to less than 41$\mu\textrm{g}$ /mg. Cholesterol concentration in egg yolk samples quantified by CTM was in accordance with that by GC method. These results suggest that CTM is an useful method for the quantification of cholesterol in egg yolk lipid and other lipids as well.

• 대한수의학회지
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• 제39권5호
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• pp.1006-1012
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• 1999

This study was conducted to evaluate the ELISA kit for measuring the level of cortisol in the urine. The CV of within-run variation and day to day variation were 0.4~2.8 and 1.8~5.7, respectively. The minimum limitation of measurement was 1ng/ml. The cross reaction was high ($CR_{50}(%)=11.4{\sim}43.2$) in prednisolone, 11-deoxycortisol, 21-deoxycortisol and predinosone. There was low and no cross reaction in other steroid. To develop the ELISA kit we measured the cortisol level in diluted urine with PBS (procedure I), extracted urine with methylene chloride (procedure II) and extracted methylene chloride-extracted urine from thin-layer chromatography (procedure III). The CV value of procedure I, II, III was 9.4~28.3%, 7.2~8.9% and 2.5~5.7%, respectively. There was significant difference between procedure I with II, and pro-cedure I with III(p < 0.01), but no difference between procedure II with III significantly(p < 0.01). The mean UCCR of urine collected through am 8 to 10 was $9.5{\pm}7.6$(0.14~28.0) in 12-month-old dog(n = 47). In this study we can measure the cortisol level in extracted urine with methylene chloride and sequential thin-layer chromatography accurately using ELISA kit.

• 한국수정란이식학회지
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• 제21권4호
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• pp.281-286
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• 2006

본 연구는 소 난관 상피 세포를 채취 체외 배양을 실시하고, 이에 착상과 관련이 있은 IL-4를 첨가하여 배양액내의 임신에 관련된 호르몬들(P4, E2, TGF-$\beta$)의 변화를 관찰함으로써, 소 난관 상피 세포와 착상과의 관계를 구명하고자 실시하였으며, 그에 따른 결과는 다음과 같다. 소 난관 상피 세포의 체외 배양시 IL-4 첨가에 의한 배양액내의 P4의 생산은 0.001 ng/ml의 IL-4를 첨가한 배양액의 P4의 농도는 배양 시간이 경과할수록 증가하는 경향을 보였으며, 24시간보다 120시간에서는 약 2배의 생산을 보여 유의적인 차이를 나타냈다(P<0.05). 0.01 ng/ml의 경우에도 0.001의 경우와 유사한 경향을 보였으나, 0.001 ng/ml의 경우보다는 다소 생산량이 낮았다. 0.1이나 1 ng/ml의 경우는 배양 시간에 따른 생산량은 다른 두 가지의 농도와 같이 배양시간 96시간까지는 증가하였으나, 배양 시간 120시간에서는 감소하였다. 소 난관 상피 세포의 체외 배양시 IL-4 첨가에 의한 배양액내의 E2의 생산은 0.001, 0.01 ng/ml 첨가시는 P4의 경우와 같이 배양 시간 72시간까지 배양 시간에 따라 생산량이 증가하여 유의적인 차이를 나타내었으며(P<0.05), 0.1 및 1 ng/ml의 경우는 배양 시간 96시간까지 증가하는 경향을 보였다. 그러나 배양 시간 120 시간에는 IL-4의 첨가 농도에 관계없이 배양 시간 24시간째의 생산량과 유사한 경향을 나타냈다. 소 난관 상피 세포의 체외 배양시 IL-4 첨가에 의한 배양액 내 TGF-$\beta$의 생산은 IL-4의 첨가 농도 및 배양 시간에 대하여 차이를 나타내지 않았으며, 유의성도 나타나지 않았다. 배양 초기에 비하여 배양시간 120시간에는 약간 생산이 낮아지는 것으로 나타나 IL-4에 의한 TGF-$\beta$의 생산은 배양 시간 96이후에는 활성이 저하하는 것으로 나타났다. 이상의 결과로 소 난관 상피 세포의 체외 배양시 IL-4 첨가는 P4 및 E2의 생산에 영향을 미치는 것으로 나타났으며, TGF-$\beta$의 생산에는 영향을 미치지 않는 것으로 나타나, IL-4는 소의 임신의 성립에 중요한 역할을 하며, 난관 상피 세포 이외의 자성 생식 기도 내에 있어서 IL-4와 관련된 기전에 대하여 더 많은 연구가 요구된다.

• 동의생리병리학회지
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• 제25권3호
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• pp.496-502
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• 2011

The present study was undertaken to investigate the effect of bee venom acupuncture therapy on the hair follicle growth by macroscopic, microscopic and immunohistochemical observation of skin 10 and 17 days after treatment. The results were as follows : Macroscopic hair follicle growth of 0.2 ml S.B.V. acupuncture treated group was more prominent than those of 0.1 ml S.B.V. acupuncture treated group and control group. Microscopic observation indicated that the hair follicle growth of control group reached anagen phase IV of hair growing cycle, and that of 0.1 ml and 0.2 ml S.B.V. acupuncture treated groups reached anagen phase VI and catagen, respectively. Immunohistochemical observations of the expression of various cytokines, enzymes and receptors in association with hair follicle cycle after local treatment of S.B.V. acupuncture therapy are as follows: Expression of fibroblast growth factor was more intense in epidermis and outer root sheath in 0.2 ml S.B.V. acupuncture treated group than that of 0.1 ml S.B.V. acupuncture treated group and control group. Expression of epidermal growth factor was more intense in bulge and outer root sheath in 0.2 ml S.B.V. acupuncture treated group than that of 0.1 ml S.B.V. acupuncture treated group and control group. Expression of c-kit receptor was more intense in epidermis, bulge and outer root sheath in 0.2 ml S.B.V. acupuncture treated group than that of control group. Expression of vascular endothelial growth factor was more intense in epidermis, bulge and outer root sheath in 0.2 ml S.B.V. acupuncture treated group than that of control group. Expression of protein kinase C-${\alpha}$ was more intense in epidermis, bulge and outer root sheath in 0.2 ml S.B.V. acupuncture treated group than control group. It is concluded that bee venom acupuncture therapy promoted the expression of various cytokines, enzymes and receptors related to the hair growth cycle for hair growth. This findings indicates that bee venom acupuncture therapy is applicable to the treatment of hair loss.

• 한방안이비인후피부과학회지
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• 제20권2호통권33호
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• pp.77-88
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• 2007

Objectives : This experimental study was performed to investigate the effects of Yeouigeumhwang-san(YUGHS) on anti-inflammation and anti-Propionibacterium acnes. Methods : The cytotoxicity of YUGHS about viability of Raw 264.7 cell was tested by using a colorimetric tetrazolium assay(MTT assay). To investigate the anti-inflammatory effets of YUGHS on LPS-induced macrophage Raw 264.7 cell, we used ELISA kit and Western blots. Inhibitory effects of YUGHS on Propionibactrium acnes were investigated by using paper disk diffusion method. Results : 1. YUGHS has no cytotoxicity under 50 ${\mu}g/ml$ concentration but over 50 ${\mu}g/ml$ has a little cytotoxicity in Raw 264.7 cell. 2. Concentration of 100 ${\mu}g/ml$ YUGHS inhibited the production of NO in the Raw 264.7 cell stimulated with LPS. 3. All concentrations of YUGHS did not inhibit the production of $TNF-{\alpha}$ in the Raw 264.7 cell stimulated with LPS. 4. All concentrations of YUGHS significantly inhibited the production of $PGE_2$ in the Raw 264.7 cell stimulated with LPS. 5. YUGHS did not inhibit the expression of COX-2 but concentration of 50 ${\mu}g/ml$ YUGHS inhibited iNOS expression in the Raw 264.7 cell stimulated with LPS. 6. YUGHS has the effect of blocking $NF-{\kappa}B$ into nucleus in LPS-induced macrophage Raw 264.7 cell 7. YUGHS did not have the inhibitory effect of Propionibactrium acnes. Conclusions : These results indicate that Yeouigeumhwang-san has anti-inflammatory effets. If further study is performed, the use of Yeouigeumhwang-san will be valuable and benificial in the therapy of acnes.

• Parasites, Hosts and Diseases
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• 제39권1호
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• pp.67-76
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• 2001

This experiment was focused on the characterization of anti- Toxoplasma monoclonal antibodies (mAbs) and the effect of mAbs on the parasite invasion of mouse peritoneal macrophages. Twenty eight mAbs including M110, M556, R7A6 and M62l were characterized by Ab titer, immunoglobulin isotyping and western blot pattern. Antibody titer (optical density) of 4 mAbs. Ml 10. M556. R7A6 and M62l. were 0.53,0.67, 0.45 and 0.39 (normal mouse serum; 0.19) with the same IgGl isotypes shown by Enzyme-linked immunosorbent assay (ELISA). Western blot analysis showed that Ml 10. M556. R7A6 and M62l reacted with the 33 kDa (p30),31 kDa (p28),43 kDa and 36 kDa protein. Immuno-gold labelling of mAbs M110, M556, R7A6 and M621 reacted with the surface membrane, dense granules and parasitophorous vacuolar membrane (PVM) , rhoptries and cytoplasm of tachyzoite, respectively. For in vitro assay, preincubation of tachyzoties with four mAbs, Ml 10, M556, R7A6 and M62l resulted in the decrease of the number of infected macrophages (p < 0.05) and the suppression of parasite multiplication at 18 h post-infection. Four monoclonal antibodies including Ml 10 (SAGI) were found to have an important role in the inhibition of macrophage invasion and T. gondii multiplication in vitro, and these mAbs may be suitable for vaccine candidates, diagnostic kit and for chemotherapy.

• 한국식품영양학회지
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• 제20권1호
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• pp.74-78
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• 2007

In vitro를 통한 혼합시료의 물 추출물과 에탄올 추출물을 시료로 사용하였고, 마우스 비장세포 증식능 및 활성 복강 대식세포에서 분비하는 사이토카인(IL-1 ${\beta}$, IL-6, TNF-${\alpha}$)의 분비능을 측정하였다. 그 결과 혼합시료의 물 추출물과 에탄올 추출물 모두 5${\sim}$250 ${\mu}$g/ml의 농도로 첨가했을 때 세포증식을 촉진하는 효과가 있는 것으로 나타났고, 물 추출물인 경우 고농도인 1,000 ${\mu}$g/ml에서, 에탄올 추출물인 경우는 500${\sim}$1,000 ${\mu}$g/ml 이상의 고농도 첨가시 세포증식이 억제되는 효과를 보였다. 복강 대식세포의 사이토카인 분비량을 측정한 결과, 혼합 물 추출물과 에탄올 추출물 모두에서 대조군보다 높은 분비량을 보였다. IL-1${\beta}$ 생성량 검색 결과, 물 추출물 투여군 10 ${\mu}$g/ml 농도 첨가에서 유의적(p<0.05)으로 높은 분비능을 보여주었고, 에탄올 추출물을 10 ${\mu}$g/ml와 100 ${\mu}$g/ml 첨가시에도 대조군보다 높은 분비량을 보였다. IL-6의 결과에서도, 물 추출물 10 ${\mu}$g/ml을 첨가했을 때 유의적(p<0.05)으로 높은 분비량을 보였고, TNF-${\alpha}$의 경우, 물 추출물의 10 ${\mu}$g/ml와 100 ${\mu}$g/ml의 농도에서 대조군에 비해 높은 TNF-${\alpha}$ 분비능을 보였다. 이상의 결과에 따르면, 혼합 물 추출물이 비장세포 증식능과 복강 대식세포에 의한 사이토카인 분비능을 상승시킴으로서 면역 기관의 주요 기능을 증진시키는 것으로 사료된다.

• 대한한방부인과학회지
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• 제19권3호
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• pp.13-24
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• 2006

Purpose : 이 연구는 MCF-7 인간 유방암 세포주에 대한 활락효령단(活絡效靈丹) 추출물의 증식억제효과, 세포독성효과, 세포사 유발효과를 확인하기 위하여 이루어졌다. Methods : MCF-7 인간 유방암 세포주는 Dulbecco's modified Eagle's medium/F12 (DMEM/F12)에 10% fetal bovine serum (FBS)와 항생제를 가하여 만든 배지를 이용하여 배양하였고 MCF-7 세포를 96-well plate에 접종한 후 다양한 농도 (0 ${\sim}$ 2000 g/ml)의 활락효령단(活絡效靈丹)이 든 배지로 처리한 후 72시간 동안 배양하였고 또한 1000g/ml의 활락효령단(活絡效靈丹)이 든 배지로 처리한 후 48, 96, 192 시간동안 배양하여 각각 MTS assay kit로 세포생존율을 측정하였다. 세포독성은 Sulforhodamine B assay 방법을 이용해 측정하였고 세포사 과정에서 MCF-7 세포에서의 caspase 활성화를 측정하기 위해 Western blotting을 수행하여 poly ADP ribose polymerase (PARP)의 절단을 확인하였다. Results : 실험결과 활락효령단(活絡效靈丹) 추출물에 의한 세포성장 및 독성효과는 시간 및 농도에 비례하는 것으로 나타났고 세포고사과정에서 작용하는 caspase의 전기질인 PARP 절단량이 활락효령단(活絡效靈丹) 처리 농도에 비례에 증가하였다. Conclusions : 활락효령단(活絡效靈丹)은 다양한 기전에 의해서 유방암 세포에 대한 억제효과를 가질 수 있는 것으로 인식할 수 있다.

• 대한핵의학회지
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• 제14권1호
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• pp.9-16
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• 1980

A 2-site immunoradiometric assay for serum ferritin was evaluated with commercially available kit. The assay required 6 hours. The slope of the standard curve kept up ideal range with the calculation of maxium binding instead of total dose until expire date. The stage II washing was more important than the stage I washing on the modified washing procedure as the bead keeping to remain in the tube. With this modified mothod, three times of tube. washing was sufficient to reduce the significant errors The measured values of serially diluted sample with standard diluting buffer was proportional to the predicted values. In the experiment of serum effect on the assay. a linear relationship from 5 to 50% serum, but beyond 50% there was reduction in measured ferritin concentration. It has a sensitivity of 2.77 ng/ml, within-assay precision (CV) of 8.0%, and between-assay reproducibility(CV) of 7.4% (mean 174.8 ng/ml).

• Asian-Australasian Journal of Animal Sciences
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• 제10권6호
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• pp.603-608
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• 1997

To improve animal production, a simple and accurate pregnancy diagnosis plays a very important role. Therefore, the purpose of this study was to develop an on-farm blood progesterone enzyme immunoassay (EIA) system for monitoring the early pregnancy in sows. Star tubes coated with mouse monoclonal anti-progesterone antibody were used for this proposed EIA system which was tested in field trials. The results could be obtained within 30 minutes either by spectrophotometry or the naked eye. Heparinized fresh blood samples collected from the ear vein of sows 17-22 days after breeding (day 0) were tested qualitatively to diagnose sows as pregnant or non-pregnant with high ( > 3 ng/ml) or low ($${{\leq_-}}3ng/ml$$) progesterone in the blood. To provided a double check data, plasma progesterone levels were also measured quantitatively by the same EIA system with some modification. Total agreement of diagnosis by the on-farm EIA kit and by farrowing or abortion from 128 tested sows was found to be 92.2% accuracy (93.1% on pregnant diagnosis and 83.3% on non-pregnant diagnosis). It was concluded that the on-farm EIA blood progesterone test is a very useful method for monitoring the early pregnancy status of sows.