• 제목/요약/키워드: phosphatase activity

검색결과 1,320건 처리시간 0.031초

Membrane associated guanylate kinase inverted-3의 AKT signaling을 통한 enterovirus replication 조절 (Membrane-associated Guanylate Kinase Inverted-3 Modulates Enterovirus Replication through AKT Signaling Activation)

  • 박진호;남궁예나;임병관
    • 생명과학회지
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    • 제26권10호
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    • pp.1182-1188
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    • 2016
  • Membrane associated guanylate kinase inverted-3 (MAGI-3)는 세포-세포 연접의 형성을 유도하는 막단백질인 membrane associated guanylate kinases (MAGUKs)의 한 종류 단백질로 140 kDa 크기를 가진다. MAGI-3는 PTEN/MMAC와 함께 협력하여 AKT/PKB의 kinase 활성을 조절하거나 MAPKs 신호전달경로로 ERK 활성을 조절로 한다. Coxsackievirus B3 (CVB3)는 가장 일반적으로 감염된 심근 세포 사멸으로 인한 바이러스성 심근염을 일으키는 enterovirus에 속하는 인간 병원체이다. 이전 연구에서 protein kinase B (PKB, 또는 AKT)와 extracellular signal-regulated kinases 1/2 (ERK1/2)의 활성은 HeLa 세포에서 CVB3 복제를 위해 필수적임이 밝혀졌다. 본 연구에서 enterovirus 복제와 AKT 신호 활성조절에서 MAGI-3의 역할을 검증하였다. MAGI-3-Flag의 발현은 CVB3 감염 후에 AKT 신호 활성과 viral capsid protein VP1의 발현을 유도하였으며 이는 MAGI-3에 의한 enterovirus 증식 조절을 보여주었다. AKT 신호는 MAGI-3 발현에 의해 enterovirus 감염과 함께 유의하게 증가하고 이것은 감염 바이러스의 증식을 활발하게 유도함을 확인하였다. 이 결과는 MAGI-3의 발현은 AKT와 ERK의 활성이 증가하고, 더 나아가 바이러스 증식과 연관이 있다는 것을 입증한다. MAGI-3는 아마도 AKT 신호 조절을 통해 enterovirus 증식 조절에 중요한 역할을 할 것으로 생각된다.

순천만 갯벌 토양의 섬유소 분해능 및 체외효소 활성 (Cellulose Degradation and Extracellulat Enzymatic Activity of the Mud Flat in Sunchon Bay)

  • 백근식;최지혁;성치남
    • 미생물학회지
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    • 제36권2호
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    • pp.130-135
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    • 2000
  • 순천만 갯벌의 3개 지점의 표층(5cm)과 심층(20 cm)토양의 물리화학적 요인과 종속영양세균의 분포, 섬유소 분해율과 체외효소의 활성을 측정하였다. 온도, 수분함량, 인산염 인 그리고 유기물함량은 각각 -1~$30^{\circ}C$, 42.1~53.1%, 0.0779~0.1961mg/g, 그리고 1.99~7.64%로 나타나었다. 섬유소 films의 분해율은 7.7%~100%/month 범위로 1~2월에 최저 그리고 8~9월에 최고의 분해율을 나타냈다. 종속영양세균의 분포는 $0.87{\times}10^6 ~ 3.6{\times}10^7 $CUFs/g dry soil의 범위에 속해Tdmu 심층의 경우 표층에 비해 낮았다. MUF-기질의 분해율로 측정한 phoshatase, $\alpha$-D-gluocosidase, $\beta$-D-glucosidase, cellobiohydrolase 활성의 변화는 각각 152.23~1779.80 nMi/hr 2.67~202.18 nM/hr, 5.03~258.26 nM/hr, 3.42~63.07 nM/hr으로 모두 하계에 상대적으로 높은 값을 나타내었다. 섬유소 분해는 수온상승이 주된 요인이었으며, 정점간, 깊이간 큰 차이는 나타나지 않았다. 체외효소의 활성은 온도 및 섬유소 분해율과 높은 양의 상관관계를 나타냈다. 종속영양세균의 분포는 섬유소 분해와 체외효소의 활성과 높은 상괸은 나타나지 않았다.

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흰쥐의 간장 및 혈청 지질농도와 임상생화학적 지표에 미치는 식이섬유 및 식이지방 급원을 달리한 식이 투여의 영향 (Effects of Different Dietary Fats and Fibers on the Lipid Concentrations of Liver and Serum and Biochemical Index in Rats)

  • 차재영;홍상식;조영수;김대진
    • 한국식품영양과학회지
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    • 제32권8호
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    • pp.1377-1384
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    • 2003
  • 본 실험에서는 Sprague-Dawley계 수컷 흰쥐의 간장 및 혈청 지질 농도와 임상생화학적 지표에 미치는 식이지방원 및 식이섬유원을 달리한 식이를 급여한 영향을 검토하였다. 실험 식이군은 5% corn oil +5% cellulose 함유 CO군, 10% corn oil +10% 차전자피 함유 CO +PH군 및 10% lard+10% 차전자피 함유 LA+PH군인 3군으로 나누었으며 실험식이와 물은 6주간 자유급여 시켰다. 체중 증가량 및 식이효율은 CO군에 비해 LA+PH군에서 유의적으로 증가하였고, CO+PH군에서는 유의적인 증가경향을 보였다. 식이 섭취량 및 장기 중량은 실험군간에 차이가 없었다. 혈청총 콜레스테롤 농도는 CO군에 비해 CO+PH군에서 유의적으로 감소하였으나, LA + PH군과는 차이가 없었다. 혈청 중성지방 및 인지질 농도는 CO군에 비해 CO +PH군 및 LA+ PH군에서 유의적으로 감소하였다. 간장 콜레스테롤 농도는 LA +PH군에서 유의적으로 감소하였다. 혈청 임상생화학적 지표 중 glutamic pyruvic transaminase 활성 은 LA+PH군에서 유의적으로 높았으며, lactate dehydrogenase 활성 및 blood urea nitrogen은 오히려 LA + PH군에서 유의적으로 감소하였다. 다른 혈청 임상생화학적 지표중 총 단백질, 알부민, 포도당, glutamic oxaloacetic transaminase 및 lactate dehydrogenase활성은 각 실험군 간에 차이가 없었다. 이상의 결과로부터 식이 차전자피에 의한 혈중 지질 농도의 저하효과는 식이지방원 lard 첨가군에 비해 corn oil 첨가군에서 더욱 현저하였다.

사람 연골하골 중간엽 줄기세포의 효율적인 골형성 유도 (Induction of Effective Osteogenesis by Mesenchymal Stem Cells from the Human Subchondral Bone)

  • 허정은;조윤제;유명철;백용현;이재동;최도영;박동석
    • Journal of Acupuncture Research
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    • 제23권5호
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    • pp.69-77
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    • 2006
  • Background : Mesenchymal stem cells (MSCs) are present in most of the tissue matrix, taking part in their regeneration when injury or damage occurs. The aim of this study was to investigate the presence of cells with pluripotential characteristics in human subchondral bone and the capacity of these cells to differentiate to osteoblast. Methods : Human subchondral bone were digested with collagenase. Isolated cells were cultured with a-MEM, 15% FBS, 10-8M dexamethasone and 50 ng/mL ascoric acid. Cells from 0 day(isolated cells), 7 day (first subculture) and 14 days (third subculture) were used to carry out phenotypic characterization experiments flowcytometry analysis with 11 monoclonal antibodies) and osteogenic differentiation experiments. Osteogenic differentiation of cells was assessment by quantification of bone extracellular matrix components by following analysis: alkaline phosphatase(ALP) stains to detect ALP activity, RT-PCR and western blot to detect osteocalcin (OCN), osteopontin (OPN) and type I collagen(Col I), and Alizarin red stains to detect calcium deposition. Results : Flowcytometry analyses showed that in our population more than 98% of cells were positive for MSC markers: SH-2(CD105, 99%), CD29 (95%), CD73 (95%). Cells were negative for hematopoietic markers (CD11b, CD34, and CD45). Furthermore, cells showed positive stain to multipotent markers such as CDl17 (c-kit) (15.1%), and CD166 (74.9%), and cell adhesion molecules such as CD54 (78.1%) and CD106 (63.5%). The osteogenic specific marker analyses showed that the culture of these cells for 7 and 14 days stimulates ALP, OCN, OPN and Col I synthesis by RT-PCR and Western blot analysis. Also, after 14 days in the culture of MSCs induces mineralization by Arizarin red stain. Conclusion : In this work, we demonstrated a new and efficient method for osteoblastic differentiation of human subchondral bone stem cells. As MSCs takes part in reparative processes of adult tissues, these cells could play an important role in osteogenesis.

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폐경 전 여성의 골밀도에 영향을 미치는 요인분석 (Factors Affecting Bone Mineral Status of Premenopausal Women)

  • 오세인;이행신;이미숙;김초일;권인순;박상철
    • 대한지역사회영양학회지
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    • 제8권6호
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    • pp.927-937
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    • 2003
  • This study was conducted to investigate dietary and other factors affecting bone mineral density (BMD) in Korean premenopausal women. Seventy-eight premenopausal women who visited the Health Promotion Center for health examinations volunteered to participate in this study. They were divided into two groups according to their bone status as shown by their T-scores: a non-osteoporotic group and a osteoporotic group. The results are as follows: The mean BMDs of the lumbar spine and femoral neck were 1.21$\pm$0.02$g/cm^2$ and 0.97$\pm$0.04$g/cm^2$, respectively. The BMD levels of the osteoporotic group were significantly lower than those of the non-osteoporotic group (p<0.001, respectively). The heights of the women in the osteoporotic group were significantly lower than those of the non-osteoporotic group (p<0.01) however, their body weights did not show any significant differences although they tended to be lower. The mean daily intake of energy was 1720$\pm$52㎉. When the nutrient intake was compared with the Korean recommended dietary allowances (RDA), calcium, Fe, vitamin A and riboflavin intakes were lower than the RDA. Their was no significant difference in the nutrient intake of the non-osteoporotic group and osteoporotic group except for the intakes of protein, fat and niacin. Their was no significant difference between the non-osteoporotic group and the osteoporotic group and all were within the normal range. However, the serum alkaline phosphatase level of the osteoporotic group was significantly higher than that of the non-osteoporotic group (p<0.001). Height measurements showed positive correlations with lumbar spine bone mineral density (LBMD, r=0.332, p<0.01) however there was no correlation with femoral neck bone mineral density (NBMD). Age, age at menarche, body weight, body mass index (BMI) and obesity showed no correlation with BMD. The BMD of the lumbar spine was significantly and positively related to the intake of niacin and vitamin C (r=0.236, p<0.05; r=0.274, p<0.05). Serum levels of calcium and phosphorus showed negative correlations with LBMD (r=-0.698, p=0.0001, r=-0.503, p=0.0001, respectively). The results suggested that the BMD of the lumbar spine was positively related to the intake of niacin and vitamin C in premenopausal women. Therefore, this study confirmed that one of the most effective ways to minimize bone loss would be have a higher intake of niacin and vitamin C rich foods and engaging habitually in physical activity may have a beneficial effect on BMD in the Premenopausal Period.

D-Galactosamine으로 유발된 간손상에 대한 민들레 열수추출물의 예방효과 (Hepatoprotective Activity of Dandelion (Taraxacum officinale) Water Extract against D-Galactosamine-Induced Hepatitis in Rats)

  • 박지영;박충무;김진주;송영선
    • 한국식품영양과학회지
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    • 제37권2호
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    • pp.177-183
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    • 2008
  • 본 연구에서는 실험동물에 민들레 열수추출물 식이를 급여한 후, GalN으로 간손상을 유발함으로써 그 예방효과를 혈액중의 생화학적 변화 및 간조직의 효소적인 변동을 통해서 규명하고자 하였다. GalN의 투여로 현저히 증가하였던 AST, ALT의 활성은 민들레 열수추출물 투여로 억제되었으나 군간의 차이는 보이지 않았고 ALP의 활성과 TBARS 함량은 3%의 추출물을 급여한 군에서 유의적인 감소를 보였다. GalN의 투여로 현격하게 높아졌던 혈중 $TNF-{\alpha}$의 농도 또한 감소하는 경향을 확인하였으나 유의적인 차이는 보이지 않았다. GalN의 투여로 억제되었던 catalase, GSH-reductase, Mn-SOD의 활성은 민들레 추출물 투여로 유의적인 회복이 관찰되었으나 GSH-px의 활성은 그 경향만을 확인할 수 있었다. 조직 검경을 통해 민들레 열수추출물의 간염 예방효과를 확인한 결과 GalN으로 인해 유발된 광범위한 간세포의 괴사와 변성, 지방변성 등이 민들레 열수추출물식이로 다소 감소하는 것을 관찰할 수 있었다. 이상의 결과로 민들레 열수추출물은 AST, ALT와 ALP의 활성 및 산화적 스트레스를 감소시키고 활성산소 해독계에 관여하는 효소의 활성을 증가시킴으로써 GalN으로 인한 간 손상을 예방하는 것으로 사료된다.

The effect of biomechanical stimulation on osteoblast differentiation of human jaw periosteum-derived stem cells

  • Lee, Ju-Min;Kim, Min-Gu;Byun, June-Ho;Kim, Gyoo-Cheon;Ro, Jung-Hoon;Hwang, Dae-Seok;Choi, Byul-Bora;Park, Geun-Chul;Kim, Uk-Kyu
    • Maxillofacial Plastic and Reconstructive Surgery
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    • 제39권
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    • pp.7.1-7.9
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    • 2017
  • Background: This study was to investigate the effect of biomechanical stimulation on osteoblast differentiation of human periosteal-derived stem cell using the newly developed bioreactor. Methods: Human periosteal-derived stem cells were harvested from the mandible during the extraction of an impacted third molar. Using the new bioreactor, 4% cyclic equibiaxial tension force (0.5 Hz) was applied for 2 and 8 h on the stem cells and cultured for 3, 7, and 14 days on the osteogenic medium. Biochemical changes of the osteoblasts after the biomechanical stimulation were investigated. No treatment group was referred to as control group. Results: Alkaline phosphatase (ALP) activity and ALP messenger RNA (mRNA) expression level were higher in the strain group than those in the control group. The osteocalcin and osteonectin mRNA expressions were higher in the strain group compared to those in the control group on days 7 and 14. The vascular endothelial growth factor (VEGF) mRNA expression was higher in the strain group in comparison to that in the control group. Concentration of alizarin red S corresponding to calcium content was higher in the strain group than in the control group. Conclusions: The study suggests that cyclic tension force could influence the osteoblast differentiation of periosteal-derived stem cells under optimal stimulation condition and the force could be applicable for tissue engineering.

임플란트 적용을 위한 하이드록시아파타이트·이산화티탄 표면의 생체적합성 평가 (A Biocompatibility Evaluation of Hydroxyapaite·Titania Surface for Dental Implant)

  • 강민경;배성숙
    • 치위생과학회지
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    • 제16권1호
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    • pp.70-76
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    • 2016
  • 이번 연구의 목적은 하이드록시아파타이트를 이용하여 블라스팅 처리한 뒤 양극산화 방법을 이용하여 하이드록시아파타이트와 이산화티탄이 복합된 표면을 만들고, 이에 대해 세포부착양상을 관찰하고 ALP 활성도와 칼슘 침착량을 측정함으로써 세포 분화능을 평가하여 표면의 생체적합성을 평가하는 데 있다. 이를 위해 실험을 진행한 결과 다음과 같은 결과를 얻었다. 접촉각 분석 결과, 복합처리된 HA+100과 HA+MAO 150은 유의하게 낮은 접촉각을 나타내었다(p<0.05). 세포부착 관찰 결과 연마 처리한 SM 시편에서는 납작한 모양으로 세포가 붙어있는 모습을 관찰할 수 있었으나, 표면 처리된 실험군에서는 세포의 모양이 시편을 따라 좀 더 길게 뻗어 부착된 모습을 관찰할 수 있었다. ALP 활성도 측정 결과 HA+MAO 150과 HA+MAO 200은 1, 2, 3주 모든 기간에서 가장 높은 ALP 활성도를 나타내었다(p>0.05). 칼슘양 측정 결과 HA+MAO 150 과 HA+MAO 200은 1, 2, 3주 모든 기간에서 가장 많은 칼슘양을 나타내었다(p<0.05). 따라서 하이드록시아파타이트 이산화티탄이 복합 코팅된 표면은 높은 표면에너지를 가지며 우수한 세포활성도를 나타내어 치과용 임플란트 표면으로 유용하게 사용될 수 있을 것이라고 생각된다.

Effect of microgrooves and fibronectin conjugation on the osteoblast marker gene expression and differentiation

  • Park, Su-Jung;Leesungbok, Richard;Ahn, Su-Jin;Im, Byung-Jin;Lee, Do Yun;Jee, Yu-Jin;Yoon, Joon-Ho;Cui, Taixing;Lee, Sang Cheon;Lee, Suk Won
    • The Journal of Advanced Prosthodontics
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    • 제7권6호
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    • pp.496-505
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    • 2015
  • PURPOSE. To determine the effect of fibronectin (FN)-conjugated, microgrooved titanium (Ti) on osteoblast differentiation and gene expression in human bone marrow-derived mesenchymal stem cells (MSCs). MATERIALS AND METHODS. Photolithography was used to fabricate the microgrooved Ti, and amine functionalization (silanization) was used to immobilize fibronectin on the titanium surfaces. Osteoblast differentiation and osteoblast marker gene expression were analyzed by means of alkaline phosphatase activity assay, extracellular calcium deposition assay, and quantitative real-time PCR. RESULTS. The conjugation of fibronectin on Ti significantly increased osteoblast differentiation in MSCs compared with non-conjugated Ti substrates. On the extracellular calcium deposition assays of MSCs at 21 days, an approximately two-fold increase in calcium concentration was observed on the etched 60-${\mu}m$-wide/10-${\mu}m$-deep microgrooved surface with fibronectin (E60/10FN) compared with the same surface without fibronectin (E60/10), and a more than four-fold increase in calcium concentration was observed on E60/10FN compared with the non-etched control (NE0) and etched control (E0) surfaces. Through a series of analyses to determine the expression of osteoblast marker genes, a significant increase in all the marker genes except type I collagen ${\alpha}1$ mRNA was seen with E60/10FN more than with any of the other groups, as compared with NE0. CONCLUSION. The FN-conjugated, microgrooved Ti substrate can provide an effective surface to promote osteoblast differentiation and osteoblast marker gene expression in MSCs.

Supplementation of Safflower Seed Powder and Extracts Enhances Bone Metabolism in Rib-Fractured Rats

  • Seo, Hyun-Ju;Moon, Kwang-Deog;Jeon, Seon-Min;Kim, Jun-Han;Cho, Myung-Sook
    • Preventive Nutrition and Food Science
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    • 제8권1호
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    • pp.46-53
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    • 2003
  • The current study investigated the effect of Korean safflower (Carthamus tinctorius L.) seed powder and its water and ethanol extracts on bone metabolism during recovery from rib-fracture induced by surgical operation in rats. 10-week-old male Sprague-Dawley rats weighing about 320 g were divided into 9 groups after arrival: 10d control (AIN 76 semi-purified diet), 10d safflower seed powder (10d SS-powder), 10d safflower seed ethanol extract (10d SS-EtOH), 10d safflower seed water extract (10d SS-$H_2O$), 20d control (AIN-76 semi-purified diet), 20d safflower heed powder (20d SS-powder), 20d safflower seed ethanol extract (20d SS-EtOH), 20d safflower seed water extract (20d SS-$H_2O$), and 20d sham-operation (20d sham), The dietary level for all the supplements was 5% based on the raw material weight. The rats were fed the experimental diets for 10 days before the rib fracture operation and for a further 10 or 20 days after the operation. A number 9 rib was fractured surgically and a sham-operation also performed. The rats were then sacrificed on the l0th or 20th day after the operation. The body weight initially decreased after the operation in all the rib-fractured groups, then gradually recovered. The concentrations of plasma osteocalcin were higher in the control group than in all the safflower-supplemented groups 10 and 20 days after the rib-fracture (p < 0.05). The bone-specific ALP (alkaline phosphatase) activity was significantly higher in the SS-EtOH group than in the other groups 20 days after the rib-fracture (p < 0.05). The level of urinary DPD (deoxypridinoline) was significantly higher in the SS-EtOH and SS-$H_2O$ groups than in the other groups 10 days after the rib-fracture. When comparing the PTH (parathyroid hormone) and calcitonin levels, the SS-$H_2O$ group exhibited the highest PTH level among the groups 10 and 20 days after the rib-fracture. Thus, it was concluded that the bone turnover during the fracture-healing period was more rapid in the rats supplemented with safflower seed powder or its fractions than in the control rats. Furthermore, the SS-$H_2O$ fraction was identified as the most effective in stimulating bone remodeling, as bone resorption and bone formation were both significantly increased during fracture healing when compared to the control group.