• 한국식품영양과학회지
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• 제28권3호
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• pp.638-645
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• 1999

This study was done to investigate effects of ${\gamma}$ irradiated beef feeding on the formation of gluta thione S transferase placental form positive(GST P+) foci, lipid peroxidation, cytochrome P450 system and microsomal glucose 6 phosphate activity in diethylnitrosamine(DEN) initiated rat hepatocarci nogenesis. Weaning Sprague Dawley male rats were fed the diet containing ${\gamma}$ irradiatied ground beef at the dose of 0, 3, 5kGy as a 20% of protein source for 8 weeks. One week after feeding, rats were intraperitoneally injected twice with a dose of DEN(50mg/kg BW). As a promoter, 0.05% phenobarbital was fed in drinking water from one week after DEN treatment until the end of experiment. At the end of 8th week, rats were sacrificed and hepatic GST P+ foci, microsomal malondialdehyde(MDA) and conjugated diene contents were determined. In addition, cytochrome P450 content and the activities of NADPH cytochrome P450 reductase and glucose 6 phosphatase were also measured. There was no significant effect by gamma irradiation on microsomal MDA content, conjugated diene, cytochrome P450 content and activities of NADPH cytochrome P450 reductase and glucose 6 phosphatase. However with DEN treatment, microsomal MDA content and conjugated diene contents were significantly changed. Cytochrome P450 content was also significantly increased while microsomal glucose 6 phophatase activity was significantly decreased with DEN treatment. However activity of NADPH cytochrome P450 reductase was not affected. An interesting finding in this study was that the number and area of hepatic GST P+ foci of the rats fed gamma irradiated beef were significantly(p<0.05) lower than those of the control. Such a lowering effect on GST P+ foci formation was highest at the dose of 3kGy than others. Overall results suggest that the consumption of low dose of gamma irradiated beef does not affect the formation of lipid peroxide, cytochrome P450 system and membrane stability.

• Radiation Oncology Journal
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• 제29권1호
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• pp.53-62
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• 2011

방사선간염은 대개 황달을 동반하지 않은 알칼리인산 분해효소(alkaline phophatase)의 상승을 특징으로 보인다. 하지만, 방사선간염 환자의 일부에서는 아미노전달효소(transaminase)의 상승을 특징적인 소견으로 보이고, 이러한 소견은 특히 간암 환자의 70~90%가 B형 간염 바이러스 보균자인 아시아 지역에서 흔하게 관찰된다. 또한 B형 간염 바이러스 보균자인 간암 환자를 방사선으로 치료했을 때 방사선간염이 더 흔하게 발생한다. 이런 사실들은 B형 간염 바이러스 보균자에서 방사선간염의 발생 기전이 비보균자에서의 그것과 다를 수 있다는 것을 시사하고, B형 간염 바이러스 보균자에서 발생하는 방사선간염의 발생기전은 B형 간염 바이러스의 재활성화와 연관이 있을 것으로 추측된다. 하지만, 현재 간암의 방사선치료 후 B형 간염 바이러스의 재활성화에 대한 연구는 미미한 수준이다. 이에 저자들은 간암 환자의 토모테라피 후 발생한 B형 간염 바이러스 재활성화 3예를 보고하고자 한다.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제35권6호
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• pp.397-402
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• 2009

The purpose of this study is to investigate the effects of alendronate and pamidronate on proliferation and the alkaline phosphatase activity of human bone marrow derived mesenchymal stem cells and to relate the results with bisphosphonate related osteonecrosis of the jaw(BRONJ). With the consent of patients with no systemic disease and undergoing iliac bone graft, cancellous bone was collected to obtain human bone marrow derived mesenchymal stem cells through cell culture. 96 well plate were prepared with a concentration of $10^4$cell/ well. Alendronate and pamidronate were added to each well with the concentration of $10^{-6}M$, $10^{-8}M$ and $10^{-10}M$, respectively. Then proliferation capacity of each well was evaluated with the cell counting kit. 24 well plates were prepared with a concentration of $10^5$cell/ml/well and with the bone supplement, alendronate and pamidronate were added with the concentration of $10^{-6}M$, $10^{-8}M$ and $10^{-10}M$, respectively on each plate. The plates were cultured for either 24 or 72 hours. Then the cells were sonicated to measure the alkaline phosphatase activity and protein assay was done to standardize the data for analysis. As the concentration of alendronate or pamidronate added to the culture increased, the proliferation capacity of the cells decreased. However, no statistical significance was found between the group with $10^{-10}M$ of bisphophonate and the control group. Pamidronate was not capable of increasing the alkaline phosphatase activity in all trials. However, alkaline phosphatase activity increased with 24 hours of $10^{-8}M$ of alendronate treatment and with 48 hours of $10^{-10}M$ of alendronate treatment. Cell toxicity increased as the bisphosphonate concentration increased. This seems to be associated with the long half life of bisphosphonate, resulting in high concentration of bisphosphonate in the jaw and thus displaying delayed healing after surgical procedures. Alendronate has shown to increase the alkaline phophatase activity of human bone marrow derived mesenchymal stem cells. However, this data is insufficient to conclude that alendronate facilitates the differentiation of human bone marrow derived mesenchymal stem cells. Further studies on DNA level and animal studies are required to support these results.

• 동의생리병리학회지
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• 제26권3호
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• pp.320-324
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• 2012

Osteoporosis is an important public health issue in postmenopausal women. It is a major public health concern and is widely believed that osteoporosis results from imbalance between bone resorption and bone formation. Recently natural products from plants have been extensively studied as therapeutic drugs to treat and prevent various diseases. Hoelen (scientific name, Poria cocos) is a mushroom that is used in traditional Chinese medicine. Hoelen exhibits anti-inflammatory activity and has a protective effect on tumor progression. However, the effect of hoelen in osteoclast differentiation remains unknown. Thus, we examined the effect of hoelen in receptor activator of nuclear factor-${\kappa}B$ ligand (RANKL)-induced osteoclast differentiation. Hoelen significantly inhibited RANKL-induced osteoclast differentiation in bone marrow-derived macrophages (BMMs) in dose dependent manner without toxicity. Also, we showed that hoelen significantly inhibited the mRNA expression of tartrate-resistant acid phophatase (TRAP) and nuclear factor of activated T cells 1 (NFATc1) in BMMs treated with RANKL. In Particular, hoelen greatly inhibited the protein expression of NFATc1. Ectopic expression of NFATc1 partially reverses hoelen-mediated inhibition of osteoclast differentiation. Taken together, our results demonstrated that hoelen may be useful treatment option of bone-related disease such as osteoporosis, reumatoid arthritis, and periodontitis.

• Clinical and Experimental Reproductive Medicine
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• 제28권1호
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• pp.33-40
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• 2001

Objective: This study was to establish the human embryonic stem (ES) cells derived from frozen-thawed blastocyst stage embryo that were destined to be discarded after five years in routine human IVF-ET program. Methods: Frozen-thawed and survived human blastocysts were treated by immunosurgery, and recovered ICM cells were cultured onto STO feeder cell layer and ICM colony was subcultured by mechanical dissociation into clumps. To identify ES cell, alkaline phosphatase staining and expression of Oct4 in replated ICM colonies were examined. Also, to examine the possibility of ES cell differentiation, retinoic acid (RA), basic fibroblast growth factor (b-FGF), nerve growth factor (NGF) were added in culture medium. In addition, to classify the specific cell type, differentiated cells were stained by indirect immunocytochemistry. Results: One ICM colony recovered from frozen-thawed six blastocysts was subcultured, continuously replated during 40 passage culture duration without differentiation. Subcultured colonies were strong positively stained by alkaline phophatase. When the expression of Oct4 in cultured ES colony was examined, Oct4b type is more clearly indicated than Oct4a one although there was not detected in embryoid body or differentiated cells. In differentiated cardiomyocytes from ES colony, cells were beaten regularly (60 times/min). In differentiated neural cells from ES colony, neurofilament (NF) 200 kDa protein, microtubule associated protein (MAP) 2 and ${\beta}$-tubulin of specific marker in neurons, glial fibrillary acidic protein (GFAP) of specific marker in astrocytes and galactocelebrocide (GalC) of specific marker in oligodendrocytes were confirmed by indirect immunocytochemistry. Also, muscle cells were detected by indirect immunocytochemistry. In addition, ES colonies can be successfully cryopreserved. Conclusion: This study suggested that establishment of human ES cells can be successfully derived from frozen-thawed blastocysts that were destined to be discarded, and obtained specific cell types (cardiomyocytes, neurons and muscle cells) through the in vitro differentiation procedures of ES cells.

• 한국균학회지
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• 제25권3호통권82호
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• pp.176-190
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• 1997

Isozyme PAGE에 의한 isozyme polymorphism 및 random amplified polymorphic DNA(RAPD) pattern의 비교를 통하여 한국에서 수집된 63개의 표고 버섯(Lentinus edodes)품종의 유연관계를 조사하여 보았다. NTSYS PC program을 통하여 비교에 사용되어진 3가지 isozyme 즉 esterase, peroxidase, acid phophatase 등은 각각 10, 7, 3개의 isozyme polymorphism type으로 나뉘어질 수 있었고, 이러한 3개의 isozyme type들은 group-average method를 이용하여 최종적으로 6개의 집단으로 구분되어질 수 있었다. 또한 RAPD를 통한 표고 품종간 유연관계 비교는 사용되어진 총 20개의 random primer 들중 3개의 primer(OPA 03, OPA 18, OPA 19)가 품종간 polymorphism을 보였고 사용되어진 전체 random primer의 PCR product들의 집단 분석을 통해 최종적으로 5개의 집단으로 구분할 수 있었다. 사용되어진 두 가지 방법 즉, isozyme polymorphism및 RAPD pattern에 의해 구분되어진 63개 표고 버섯 품종의 subgroup level에서의 절대적인 유사성은 관찰할 수 없었다. Yii-Mattila(1996) 등에 의하면 품종간 비교시 isozyme analyses에서와는 달리 RAPD analyses에서는 많은 수의 strain-specific band들이 얻어질 수 있고 이것이 두 방법간의 유사성을 관찰하지 못하게 하는 원인이라고 보고하였다.

• 한국가축번식학회지
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• 제8권1호
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• pp.46-55
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• 1984

The purpose of the experiment was to clarify morphologically normal growth pattern of the ductus deference in accordance with the sex maturity of meat-type cockerels. 1. Diameter of lumens in u, pp.r, mid and lower parts of ductus deferens, the most conspicuous enlargement of lumen was observed in the lower part. Heights of epithelial layers of ductus deferens showed abrupt growth at 12 weeks of age with subsequent gradual growth in all the part of u, pp.r, mid and lower, and heights of those at 30 weeks were a, pp.oximately 4 times as large in the u, pp.r and mid parts and 5 times as large in the lower part in contrast to those at 4 weeks of age. Thickness of muscular layer of ductus showed gradual growth in contrast with the diameter of lumen and height of epithelial layer, showing 1.3 times as large in the u, pp.r part, 1.6 times in the mid part and 1.9 times in the lower part at 30 weeks of age in contrast to the thickness at 4 weeks of age. 2. Within 10 weeks after hatching, lining cells of ductus deferens were mainly composed of round cells and columnar cells in simple columnar epithelium. During 10th to 20th week, the lining cells were mainly composed of high columnar cells and round cells in pseudostratified epithelium. From 22nd week, the lining cells were composed of pseudostratified columnar cells. Whereas round cells disa, pp.ared gradually. Enlargement of lumen and pooling of sperms in ductus deferens coincided with the maturation of seminiferous tubules. 3. In simple correlation between the values of testis weight and the values from various measurements in the ductus deferens, there was significant correlation coefficient with each other. 4. In the India ink absorption test, India ink granules were not absorbed on the epithelium of the ductus deferens, but the granules reactive to acid phosphatase a, pp.ared in a line on the free border of each parts of the ductus deferens. The granules reactive to alkaline phosphatase were noted on the luminal border of ductus deferens mainly, but weak reaction showed than acid phophatase were a, pp.ared. The granules reactive to PAS were a, pp.ared mostly near on the free border of hte epithelial cells of ductus deferens. 5. Number of sperm, Indes of sperm vitality and MRT in the different parts of ductus deferens were tended to be somewhat dominant in the mid and lower parts than in u, pp.r part, even though not significant in the statistical analysis. Ratio of sperm abnormality was tended to be relatively high in the u, pp.r part too, and in the sperm of abnormality blunted head was less in number significantly in the mid and lower part than in the u, pp.r part.

• 대한치과교정학회지
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• 제31권3호
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• pp.357-367
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• 2001

당뇨병 흰쥐를 대상으로 치조골의 교체(turnover)와 치아 이동 중 치조골의 흡수와 형성의 활성을 비교 관찰하기 위하여 8주령의 웅성 Sprague-Dawley계 흰쥐를 정상대조군(N), 당뇨군(D), 정상-치아이동군(N-tm), 당뇨-치아이동군(D-tm)으로 나누고, 각 실험군은 치아이동 실험 1일, 3일, 7일, 14일의 소군으로 나누었다. 당뇨병은 치아이동 실험 18일전에 스트렙토조토신(50mg/체중 kg)을 꼬리정맥에 단 회 주사하여 유발하였다. 치아이동은 상악 제1대구치를 초기 교정력 40그램으로 근심 이동시켰다. 상악 제1대구치 주위 치조골을 대상으로 골형성 징후표지로서 치조골 alkaline phosphatase(ALP)와 osteocalcin(OC)을, 골흡수 징후표지로서 acid phosphatase (ACP)와 tartrate-resistant acid phophatase(TRAP)을 정량 분석하였으며, D군과 N군에서는 이들의 혈중 농도를 정량 분석하였다. 1. D군의 혈청 TRAP농도와 혈청 OC농도는 모든 실험 경과군에서 N군보다 감소하였고(p<0.05), 당뇨기간이 길어질수록 더 많이 감소한 것으로 나타났다. 또한 치조골 흡수 징후표지는 감소하는 경향이었으며 골형성 징후표지도 실험기간이 경과함에 따라 더 많이 감소하는 것으로 나타났다. 2. N-tm군의 치조골 ACP농토와 TRAP농도는 N군과 비교하여 1일과 3일군에서 가장 높았으며(p<0.01), 그 이후 감소경향을 나타냈고, 치조골 OC함량은 3일, 7일 14일군에서 N 군보다 높았다(p<0.001). 그러나 D-tm군에서 치조골 ACP와 TRAP농도는 정상 N군에 비하여 1일에는 낮으나 7일, 14일 경과군에서 가장 높았고(p<0.001), 실험 경과에 따라 계속 증가 하였으며(p<0.001), 치조골 ALP와 OC 함량은 3일 경과 후부터 증가하지만 정상 N군과 유의차가 없었다. 3. 당뇨-치아이동군의 치아이동량은 정상-치아이동군보다 많은 것으로 나타났다. 이러한 실험 결과는 당뇨병의 전신 골격과 치조골은 골 흡수 활성이 억제되어 있으며 골형성 활성은 당뇨 지속시간이 길어질수록 더 많이 억제되는 느린 골교체를 하며, 교정력에 의한 치조골 흡수활성이 처음 높아지는 시기는 정상치아이동시보다 늦고 더 오랬 동안 지속되며 치조골 형성활성은 낮고, 치아 이동량은 증가하며 이동중 치아동요도가 증가함을 시사한다.

• 미생물학회지
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• 제52권1호
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• pp.10-17
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• 2016

항체의 특이적 결합을 분석하는 효소면역분석법은 항원의 탐지를 위해 주로 horseradish peroxidase (HRP) 또는 alkaline phosphatase (AP) 등의 효소를 사용한다. 이때 효소를 주로 화학적으로 항체에 결합시켜 사용하게 되는데 이 과정이 복잡하며 불규칙하게 일어나서 항체 및 효소의 기능을 감소시키게 된다. 또한 대부분의 효소면역분석법에서는 주로 일차 항체의 항원결합을 탐지하기위해 이차 항체를 사용하는데, 즉 이차 항체에 결합한 효소의 기질발색에 의해 일차 항체의 항원결합을 탐지하므로 이차 항체가 요구 되어질 뿐만 아니라 이차 항체의 일차 항체에 대한 반응을 위한 부가적인 배양시간이 필요하다. 더욱 더 중요한 것은 이차 항체만의 비특이적 항원 결합 역시 제거되어져야 한다. 본 연구에서는 대장균의 genomic DNA로부터 PCR을 통해 alkaline phosphatase 유전자(Sadeghi et al., 2008)를 증폭 분리한 다음 이를 TRAIL (tumor necrosis factor ${\alpha}$ related apoptosis induced ligand) receptor인 death receptor 4 (DR4)에 특이적으로 결합하는 hAY4 single-chain Fv (ScFv)에 융합시킨 재조합 ScFv-AP 형태로 대장균에서 발현시켜 정제하였다. 정제된 hAY4 ScFv-AP는 SDS-PAGE에서 단량체(monomer) 분자량인 73.8 kDa을 나타내었다. 그러나 size-exclusion chromatography(SEC)에서는 147.6 kDa을 나타내는 결과를 통해 hAY4 ScFv-AP는 AP의 자연적인 비공유결합에 의해 이량체(dimeric form)형성이 유도되어짐을 확인하였다. 또한 ELISA, Western blot 그리고 immunocytochemistry에서 이차 항체 없이 일차 항체 hAY4 ScFv에 직접 융합된 AP의 기질발색에 의해 ScFv 일차 항체의 특이적 항원결합을 나타내었다. 요약하면 hAY4 ScFv와 대장균의 alkaline phosphatase 유전자를 융합시켜 대장균에서 수용성 형태로 성공적으로 정제하였으며 정제된 ScFv-AP 융합단백질은 ELISA, Western blot 및 immunocytochemistry에서 항원결합력을 나타냈으며 또한 구매에 따른 고비용, 부가적인 배양시간 및 비특이적 결합에 의한 오류 등의 문제점을 갖는 이차 항체를 사용하지 않고 직접적인 항원결합력을 나타내었다.

• 대한소아치과학회지
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• 제28권2호
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• pp.238-246
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• 2001

본 실험은 탁월한 골유도능으로 관심의 대상이 되고 있는 BMP의 세포내 신호 전달자로 알려진 Smad 1과 Smad 5가 조골세포 초기 분화표지인자인 ALP 유전자의 발현에 미치는 영향 및 그 조절기전을 알아보고자 하였다. BMP 처리 없이도 Smad에 의해 ALP가 발현되는가를 알아보기 위해 Smad 1과 Smad 5가 각각 stably transfection된 C2C12 세포를 3일간 배양후 histochemical assay를 하였고, Smad 1과 Smad 5의 expression vector와 ALP promoter vector를 transient co-transfection한 후 ALP promoter activity를 측정하였다. Smad에 의한 BMP의 효과를 알아보기 위해서 100ng/ml의 BMP-2를 처리한 군과 처리하지 않은 군으로 나누어 세포를 배양한후 ALP 유전자의 발현을 northern blot analysis로 확인 하였다. Smad가 ALP 유전자의 발현을 직접적으로 조절하는가를 알아보기 위해서는 단백질 합성억제제인 cycloheximide를 전처리하여 ALP 유전자의 발현을 northern blot analysis하였다. 이상의 실험결과 다음과 같은 결론을 얻었다. $\cdot$ Smad 1과 Smad 5가 과발현된 세포에서는 BMP 처리없이도 ALP가 발현된다. $\cdot$ Smad 1과 Smad 5가 과발현된 세포에서 BMP 처리후 ALP 발현 증가율이 대조군 보다 현저히 높게 나타나 Smad가 BMP 효과를 증가시킨다는 것을 알 수 있다. $\cdot$ Smad는 새로운 단백질의 합성을 통해 ALP 유전자를 발현시킨다.