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GAPDH, β-actin and β2-microglobulin, as three common reference genes, are not reliable for gene expression studies in equine adipose- and marrow-derived mesenchymal stem cells

  • Nazari, Fatemeh;Parham, Abbas;Maleki, Adham Fani
    • Journal of Animal Science and Technology
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    • 제57권5호
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    • pp.18.1-18.8
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    • 2015
  • Background: Quantitative real time reverse transcription PCR (qRT-PCR) is one of the most important techniques for gene-expression analysis in molecular based studies. Selecting a proper internal control gene for normalizing data is a crucial step in gene expression analysis via this method. The expression levels of reference genes should be remained constant among cells in different tissues. However, it seems that the location of cells in different tissues might influence their expression. The purpose of this study was to determine whether the source of mesenchymal stem cells (MSCs) has any effect on expression level of three common reference genes (GAPDH, ${\beta}$-actin and ${\beta}2$-microglobulin) in equine marrow- and adipose-derived undifferentiated MSCs and consequently their reliability for comparative qRT-PCR. Materials and methods: Adipose tissue (AT) and bone marrow (BM) samples were harvested from 3 mares. MSCs were isolated and cultured until passage 3 (P3). Total RNA of P3 cells was extracted for cDNA synthesis. The generated cDNAs were analyzed by quantitative real-time PCR. The PCR reactions were ended with a melting curve analysis to verify the specificity of amplicon. Results: The expression levels of GAPDH were significantly different between AT- and BM-derived MSCs (p < 0.05). Differences in expression level of ${\beta}$-actin (P < 0.001) and B2M (P < 0.006.) between MSCs derived from AT and BM were substantially higher than GAPDH. In addition, the fold change in expression levels of GAPDH, ${\beta}$-actin and B2M in AT-derived MSCs compared to BM-derived MSCs were 2.38, 6.76 and 7.76, respectively. Conclusion: This study demonstrated that GAPDH and especially ${\beta}$-actin and B2M express in different levels in equine AT- and BM-derived MSCs. Thus they cannot be considered as reliable reference genes for comparative quantitative gene expression analysis in MSCs derived from equine bone marrow and adipose tissue.

포도당(葡萄糖) 이성화(異性化) 효소(效素)에 관(關)한 연구(硏究) -제3보(第三報) 분리(分離) 선정(選定)된 균주(菌株) 효소(酵素)의 작용(作用)에 미치는 금속염(金屬鹽)의 영향(影響)에 대(對)해서- (Studies on the Glucose Isomerizing Enzyme -Part III The Effects of Metal Ions on the Reaction of K-17 Strain's Glucose Isomerizing Enzyme-)

  • 서정훈;김종규;기우경;이인구;권태종;우두리
    • Applied Biological Chemistry
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    • 제11권
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    • pp.55-61
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    • 1969
  • 본(本) 연구(硏究)에서 사용(使用)한 K-17 균주(菌株)의 효소생성(酵素生成)및 그의 금속염(金屬鹽) 등(等)에 대(對)한 영향(影響)등(等)을 요약(要約)하면 대략(大略) 다음과 같다. 1. 본(本) 균주(菌株)는 Glucose Isomerizing Enzyme 생성(生成)에 있어서 Xylose를 절대적(絶對的)으로 요구(要求)하지 않는다. 2. 본(本) 효소(酵素)는 Free Incubation 상태(狀態)에서는 $75^{\circ}C$, 30시간(時間)의 가열(加熱)로서 대부분(大部分)의 효소(酵素)가 실활(失活)된다. 3. 금속(金屬) Ion 중(中) $Mg^{++}$는 효소작용(酵素作用)에 있어 일종(一種) 의 Activator로 작용(作用)하며 $Co^{++}$는 Activator로 서의 작용(作用)은 없고 효소(酵素)의 열(熱) 실활(失活)을 보호(保護)하는 작용(作用)을 나타낸다. 4. 본(本) 효소작용(酵素作用)에 있어서 Glucose의 용매(溶媒)로서는 Phosphate Buffer가 가장 우수(優秀)한 결과(結果)를 나타내었다. 5. 본(本) 균주(菌株)의 효소(酵素) 생성(生成) 배지(培地)로서는 탄소원(炭素源)으로서 주어지는 당류(糖類)의 혼용(混用)이 단용(單用)에 비(比)해 균체(菌體)의 수량(收量)이나 효소능(酵素能)이 다같이 높아짐을 알게 되었다.

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분포반사기 레이저 다이오드와 광흡수 변조기가 집적된 소자의 제안 및 해석 (Proposan and Analysis of DR(Distributed Reflector)-LD/EA(electro-absorption)­Modulator Integrated Device)

  • 권오기;심종인
    • 한국광학회지
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    • 제9권5호
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    • pp.333-341
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    • 1998
  • 본 논문에서는 기존의 분포궤환형 레이저 다이오드와 광흡수 변조기(DFB-LD/EA­ MOD.)가 집적된 소자의 낮은 광효율과 수율의 문제점을 해결하고, 동시에 우수한 chirp특성을 나타내는 분포반사기 레이저 다이오드와 광흡수 변조기 (DR-LD/EA­MOD.)가 집적된 소자를 제안하였다. DR-LD/EA­MOD. 집적소자는 제작상으로 DR-LD/EA­MOD. 소자와 비교해서 선택 MOVPE성장시 SiO2 마스크 폭만 조절하는 것 외엔 거의 동일하므로 실용적 측면에서도 우수한다. 집적소자의 정특성 및 동특성을 해석하기 위하여 시간의존성을 갖는 전달 메트릭스방법, 활성층내의 비율 방정식, 변조기의 QCSE효과를 고려한 Schrodinger 방정식을 동시에 풀이하였다. 1.55$\mu\textrm{m}$ DR-LD/EA­MOD. 집적소자와 종래 사용중인 1.55$\mu\textrm{m}$ DR-LD/EA­MOD. 집적소자의 성능을 비교 분석하여, LD에 동일전류를 주입할 경우 광변조기의 on상태의 광출력이 약 30%이상 향상되고, 동일 광변조기의 잔류단면반사율에 대해 광출력의 ripple이 적고, 동적파장 천이량이 약 50%이상 줄어듦을 알 수 있었다. 또한 DR-LD/EA­MOD. 소자는 시분할 파장특성은 광 펄스의 leading edge에서 blue-shift, falling edge에서 red-shift 특성이 기대되었다. 이는 일반적인 단일모드 광섬유를 사용하여 광 펄스를 전송할 경우 전송시 펄스 폭이 좁아지는 효과를 주어 이로 인해 전송대역폭을 크게 향상시킬 수 있음을 의미한다.

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비골 골절 교정 후 Rolled Silastic Sheet를 이용한 비강 충진의 효과 (The Effect of Nasal Packing with Rolled Silastic Sheet after Closed Reduction of Nasal Bone Fracture)

  • 손경민;양정열;김규보;한윤주;천지선
    • Archives of Plastic Surgery
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    • 제38권5호
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    • pp.602-608
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    • 2011
  • Purpose: Nasal bone fracture is the most common type of facial bone fracture and most of nasal bone fracture is combined with septal fracture frequently. Nasal septum is important to support the distal nose and to maintain the nasal airway. But nasal septal fractures are usually unrecognized and untreated at the time of operation. Recently, various materials were using for nasal packing after closed reduction, however these materials are not focused on the correction of nasal bone and nasal septal fracture and many patients are suffered from nasal packing materials. Thus, the purpose of this study is to compare routine packing materials and rolled silastic sheet with respect to postoperative effect of correction of nasal bone fracture and discomfort of nasal packing materials. Methods: We examined 320 patients treated nasal bone fracture from January 2008 to December 2010. For Group I (n=92), $Merocel^{(R)}$ was used for nasal packing, for Group II (n=152) vaseline gauze was used, and Rolled silastic sheet (RSS) with vaseline gauze packing (VGP) was used for Group III (n=76). Under the general anesthesia, all patients were operated by closed reduction and nasal packing was done using three kinds of packing materials. At the time of postoperative 7 days, packing material was removed and studied for pt's satisfaction and postop. complications. Results: In patients with RSS with VGP, the complaints (nasal obstruction, foreign compressive sensation and discomfort during food ingestion) of keeping the nasal packing were decreased ($p$ <0.05) and the postoperative complication (deviation) were decreased comparing to vaseline gauze packing and $Merocel^{(R)}$ packing, however, these differences were not statistically significant ($p$ >0.05). Conclusion: Postoperative nasal packing with RSS with VGP was more comfortable to the patients and it was more effective method to correct the nasal bone fracture and nasal septal fracture.

화상 반흔에서 생긴 궤양을 동반한 이소성 석회화의 치료 (Treatment of Heterotopic Calcification with Ulceration in Burn Scar)

  • 강상구;이종욱;고장휴;서동국;최재구;장영철
    • Archives of Plastic Surgery
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    • 제37권4호
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    • pp.415-420
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    • 2010
  • Purpose: Heterotopic calcification is a pathological event in which deposits of calcium salts build up, usually in the joint area or soft tissues. It can occur under many conditions and in some rare cases may develop in burn scars. In particular, ulcerations in burn scars accompanied by heterotopic calcification are difficult to treat through conservative treatment. This study reports methods for accurate diagnosis and adequate treatment of ulceration in burn scars accompanied by heterotopic calcification. Methods: Fourteen patients who visited our hospital from March 2008 to January 2010 were subjected to this study. Their sex, age, modes of burn, degree, TBSA (%), ulcerated area, the time of occurrence of the ulcerations in the burn scars were investigated. In addition, radiological examination and biopsy was performed to diagnose heterotopic calcification. Results: Among the 14 cases, 6 were male and 8 were female. The average age of the patients was 48.2 (27 - 69 yrs). As for the mode of burn, 11 were flame burns and 3 were scalding burns. The average time of occurrence of the ulcerations in the burn scars was 4.5 months. The ulcerated areas were situated in the legs in 12 cases, arms in 1 case, and torso in 1 case. The diagnosis was confirmed through X-ray and biopsy, and skin graft was performed after wide excision. Conclusion: Diagnosis of the ulceration in burn scars accompanied by heterotopic calcification is possible through radiological and pathologic studies. Surgical treatment is the most reliable method of treatment, and we chose to perform skin graft after wide excision. Also, we learned that the complete removal of the calcified tissue and the inflammatory fibrotic tissues is crucial in preventing recurrence. Also, in contrast to Marjolin's ulcer, heterotopic calcification had a small size, little or no granulation tissue, and lacked fungating type ulceration. Therefore, favorable prognosis could be achieved through adequate treatment.

피부 제거 없이 시행한 높은 쌍꺼풀의 교정 (Correction of High Fold without Skin Excision)

  • 오흥찬;윤동주;강철욱;최치원;최수종;배용찬
    • Archives of Plastic Surgery
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    • 제36권5호
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    • pp.649-653
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    • 2009
  • Purpose: Double - eyelid operation is one of the most common cosmetic operations among Korean patients. In such operations, however, the complexity of and individual differences in the patients' anatomical structures may cause various complications, such as asymmetry, retraction of the eyelid, and the occurrence of a high fold. A high fold occurs frequently, and its correction is not very simple. Many methods have been developed to correct it, and among these, the operation involving the excision of the skin between the previous double - eyelid line and the new double - eyelid line is usually selected by plastic surgeons. In many cases, however, patients have insufficient eyelid skins for this operation. In this study, the authors introduce an operation procedure for high - fold correction that does not involve skin excision Methods: From June 2005 to June 2009, 246 cases were treated with this procedure. After the incision of the new double - eyelid line, dissection was done between the previous scar tissue and the levator aponeurosis. Then the orbital septum, orbital fat or the retro - orbicularis oculi fat was slid down and sutured with a tarsal plate. Such sutures were repeated at four to five points, including the lateral and medial limbus, to prevent the reattachment of the previous scar and to create a new double - eyelid line at the end of the orbital septum. Results: Most of the high - fold patients were satisfied with the procedure described above. Their previous scar was hidden under the new double - eyelid line after the operation. In the six cases, the scar was visible in the patients who had a very high and deep inner line. As such, scar revision was undertaken three months after the operation. It is known that scar revision is also required after an operation involving skin excision in the case of a very high inner - eyelid line. Conclusion: This method is an appropriative procedure for high - fold correction for patients who have insufficient upper - eyelid skin.

가상 n형 폴리아닐린의 제조 및 전기화학적 특성평가 (Preparation of pseudo n-type Polyaniline and Evaluation of Electrochemical Properties)

  • 김래현;최선용;정건용
    • 멤브레인
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    • 제13권3호
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    • pp.162-173
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    • 2003
  • 폴리아닐린(polyaniline, PANI)과 도판트인 camphorsulfonic acid(CSA), dodecyl benzene sulfonic acid(DBSA)와 의몰비 변화에 따라 가상 n형 PANI을 제조하였다. FT-IR측정으로부터 도핑유무를 확인하였고, indium thin oxide(ITO)에 코팅하여 제조한 전극에 대해, 순환전압전류법과 교류임피던스법을 이용하여 도판트의 영향을 조사하였다. FT-IR과 순환전압전류법으로부터, 제조된 전극이 양이온의 도핑-탈도핑이 일어나는 가상 n형의 특성을 가짐을 확인하였다. 순환전압전류법에서 산화-환원 피크전류값은 PANI/DBSA에 비하여 PANI/CSA가 약 5 배정도 더 큰 결과를 보였다. 교류임피던스법으로부터, 두 전극 모두 이상적인 Randles의 등가회로와 유사한 거동을 보였다. 전하이동저항은 PANI/CSA에서 $1.14~1.09 k{\Omega}$으로 거의 일정한 값을 보였고, PANI/DBSA는 DBSA 몰 비에 증가에 따라 $27.73{\sim}8.37K{\Omega}$으로 감소하여 나타났다. 이중층용량 또한 PANI/CSA는 $13.47{\sim}14.59 {\mu}F$으로 거의 일정하였으나, PANI/DBSA는 DBSA 몰 비 증가에 따라 $0.49{\sim}l.20 {\mu}F$으로 증가를 보였다. 결과적으로 PANI/CSA의 전기적 특성이 더 좋았으나, 도판트의 몰 비 증가에 따른 특성은 PANI/CSA 전극은 거의 일정하였고, PANI/DBSA 전극은 전기적 활성이 좋아짐을 알 수 있었다.

북방산개구리(Rnan dybowskii )유생의 포식자회피 행동에 대한 FNW와 FNW-Ag 복합체의 영향 (Effects of FNW and FNW-Ag on the Antipredator Behavior of Dybowski's Frog (Rana dybowskii) Larvae)

  • 김은지;고원배;한얼;고정원;정훈
    • 한국습지학회지
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    • 제20권3호
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    • pp.272-280
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    • 2018
  • 기존의 독성평가는 생물의 생존가능성에 영향을 줄 수 있는 이상행동반응에 대한 평가를 거의 하지 않았다. 따라서 본 연구에서는 행동학적 이해를 바탕으로 양서류 유생들의 생존가능성과 직결되는 포식자회피반응 행동을 이용하여 fullerene nanowhisker(FNW)와 nanowhisker-silver nanoparticle composites(FNW-Ag복합체)의 독성을 평가해 보았다. 우리는 나노물질에 노출되지 않는 그룹과 5가지 농도에 따른 나노물질에 노출되는 그룹(FNW $10{\mu}g/ml$, $100{\mu}g/ml$, $500{\mu}g/ml$과 FNW-Ag복합체 $10{\mu}g/ml$, $50{\mu}g/ml$)으로 나누어 실험하였다. 그 결과, FNW-Ag복합체 $50{\mu}g/ml$농도에 노출된 유생들을 제외하고는 평소 활동량에는 차이가 없었지만, FNW $10{\mu}g/ml$농도에 노출된 유생들을 제외하고는 모두 포식자인식 반응에 따른 활동변화에서는 통계적으로 유의한 차이가 나타났다. 즉, 나노물질노출 유무는 평소 움직임에는 영향을 주지 않지만, 행동학적 이해를 바탕으로 한 분석결과 포식자회피반응에 영향을 준다는 것을 알 수 있었다. 따라서 생물을 대상으로 하는 독성평가연구에 행동학적 분석이 이루어져야 할 것으로 판단된다.

독수리(Aegypius monachus)의 흡입마취에서 Capnography를 이용한 호기말 이산화탄소분압 모니터링 (Capnographic Monitoring of End-Tidal Partial Pressure of Carbon Dioxide in Anesthetized Cinereous vultures (Aegypius monachus))

  • 박원규;석성훈;박세진;이승용;김영기;황재민;정동혁;이희천;연성찬
    • 한국임상수의학회지
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    • 제32권4호
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    • pp.380-384
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    • 2015
  • The relationships between end-tidal partial pressure of carbon dioxide ($P_{ET}CO_2$) and arterial partial pressure of carbon dioxide ($P_aCO_2$), and between $P_{ET}CO_2$ and blood pH in isoflurane-anesthetized cinereous vultures were evaluated. Nine cinereous vultures (weighing 7.3-9.5 kg) were anesthetized via mask with isoflurane, intubated, and connected to a mechanical ventilator. By mechanical ventilator, respiratory rate was altered to achieve a $P_{ET}CO_2$ randomly selected among 30 to 35 mmHg. $P_aCO_2$ and blood pH were determined in serial arterial samples and compared with concurrent $P_{ET}CO_2$ measured with a sidestream capnography. Thirty six samples, nine cinereous vultures were tested 4 times each, were used to determine correlations coefficients between $P_aCO_2$ and $P_{ET}CO_2$, and between $P_{ET}CO_2$ and blood pH. Strong correlations were observed between $P_aCO_2$ and $P_{ET}CO_2$ (r = 0.951, P = 0.000, y = 1.679x - 30.236, $R^2=0.905$) as well as between $P_{ET}CO_2$ and blood pH (r = -0.728, P = 0.000, y = -0.024x + 8.389, $R^2=0.530$) with Pearson's correlation coefficient and simple linear regression. These results suggest that the capnography used provided a sufficiently accurate estimation of $P_aCO_2$ and blood pH. Thus, the monitoring of $P_{ET}CO_2$ can be useful method to prevent hypoventilation for cinereous vultures receiving mechanical ventilation.

논발생(發生) 담수조류(湛水藻類)에 관(關)한 연구(硏究) - V. 시비반응(施肥反應) 및 방제(防除) (Fresh-water Algae Occurred in Paddy Rice Fields - V. Fertilizer Response and Chemical Control)

  • 이한규;박재읍;류갑희;이정운;박영선
    • 한국잡초학회지
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    • 제13권1호
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    • pp.1-6
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    • 1993
  • 논에 발생(發生)하는 주요(主要) 조류(藻類)의 번식(繁殖)에 미치는 시비(施肥) 및 염분농도(鹽分濃度)의 영향(影響)과 약제방제법(藥劑防除法)을 구명(究明)하기 위한 시험(試驗)을 1991년 수행하여 다음과 같은 결과(結果)를 얻었다. 1. 남조류(籃藻類)인 흔들말(Oscillatoria)의 발생량(發生量)은 질소비료(窒素肥料)의 증시(增施)에 따라 증가(增加)하였고 녹조류(綠藻類)인 그물말(Hydrodictyon)의 발생량(發生量)은 인산비료(燐酸肥料)의 증시(增施)에 따라 증가(增加)하였다. 2. 그물말은 질소비료(窒素肥料)에 극(極)히 민감(敏感)하여 질소비료(窒素肥料)의 증시(增施)에 따라 민감장해(敏感障害)를 일으켰다. 3. 유그레나도 다량(多量)의 질소비료(窒素肥料) 시용(施用)으로 번식장해(繁殖障害)를 보였으나 그물말에 비(比)하여 민감(敏感)하지는 않았다. 4. 흔들말, 클라도포라 및 깃돌말(Navicula)은 내염성(耐鹽性)을 보였다. 5. Piperophos/dirnethametryn은 정체성(靜滯性) 녹조류(綠藻類)인 그물말과 부유성(浮遊性) 단세포(單細胞) 녹조류(綠藻類) 및 유그레나의 방제(防除)에 효과(效果)가 높았고 propineb와 류산동(硫酸銅)도 그물말에 대(對)하여 높은 방제효과(防除效果)가 인정(認定)되었다.

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