• Reproductive and Developmental Biology
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• 제37권3호
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• pp.97-102
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• 2013

The objective of this study was to develop of semen transport system for cryopreservation and fertility in bull sperm. The ejaculated semen were diluted with Triladyl containing 20% egg-yolk for transportation. Diluted semen was transported by three methods that there were wrapping tissue (Tissue), sinking under $30^{\circ}C$ water (Water) and sinking between warm water and air (Air) methods. Semen was transported within 2 hours in $0.3^{\circ}C$. For this study, the freezing of diluted semen were added with Triladyl containing 20% egg-yolk. And frozen-thawed sperm were estimated with SYBR14/PI double stain for viability, FITC-PNA/PI double stain for acrosome reaction analysis and Rhodamine123 double stain for mitochondrial intact assessment. In results, live sperm (SYBR+/PI-) in Air treatment group ($43.3{\pm}4.7%$) was significantly (p<0.05) higher than other treatment groups (Tissue: $16.3{\pm}2.7%$ and Water: $27.5{\pm}3.1%$), dying sperm (SYBR+/PI+) in Air treatment group ($55.6{\pm}4.7%$) was significantly lower than other treatment groups (Tissue: $77.6{\pm}3.2%$ and Water: $67.6{\pm}3.3%$) (p<0.05). Acrosome reaction in Air treatment group ($0.2{\pm}0.1%$) within live sperm (PI negative region) was significantly (p<0.05) lower than other treatment groups (Tissue: $0.7{\pm}0.2%$ and Water: $0.5{\pm}0.1%$), the acrosome reaction in Air treatment group ($28.6{\pm}2.8%$) within all sperm also was significantly lower than other treatment groups (Tissue: $44.2{\pm}1.8%$ and Water: $36.2{\pm}2.0%$) (p<0.05). And mitochondrial intact in Air treatment group within live ($97.1{\pm}0.4%$) and all ($61.9{\pm}3.3%$) sperm were significantly higher than other treatment groups (Tissue: $85.2{\pm}3.3%$, Water: $87.8{\pm}2.9%$ within live sperm and Tissue: $49.28{\pm}3.7%$, Water: $42.0{\pm}3.1%$ within all sperm) (p<0.05). Therefore, we suggest that transportation by sinking method between warm water and air was beneficial to improvement of fertility in frozen-thawed in bull semen.

• Animal cells and systems
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• 제15권4호
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• pp.287-294
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• 2011

The accelerated cooling rate associated with vitrification reduces injuries attributed to cryopreservation and improves the post-freezing developmental competence of vitrified embryos. In this study, embryos were vitrified and warmed and morphologically evaluated for their development to blastocysts. Survival rates between the fresh ($96.7%{\pm}3.8%$) and vitrified embryos ($90.7%{\pm}5.1%$) did not differ significantly (P>0.05). The mitochondrial membrane potential of fresh control cells measured by 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolyl carbocyanide iodide staining was similar to that of cryoprotected and vitrified embryos. Mitochondrial staining with rhodamine 123 did not differ among the fresh, cryoprotected, and vitrified embryos. Moreover, the distribution of $H_2O_2$, assessed by 2',7'-dichlorodihydrofluorescein diacetate staining, did not differ among the groups. The results showed that the developmental rate did not differ significantly among the fresh ($87.8%{\pm}11.3%$), cryoprotected ($83.2%{\pm}7.6%$), and vitrified 2-cell embryos ($75.8%{\pm}14.2%$). The mean number of the inner cell mass (ICM), trophectoderm (TE), and apoptotic cells was counted and statistically compared, and although the number of ICM and TE was decreased in the cryoprotected and vitrified embryos, there were no significant differences among the groups (P>0.05). During the cultivation period, randomly selected blastocysts from each group were stained using either 4',6-diamidino-2-phenylindole and bisbenzimide or the terminal deoxynucleotidyl transferase mediated dUTP nick-end labeling technique. The incidence of apoptosis appeared to be almost identical in all the groups. Droplet vitrification could subsequently lead to high survival and developmental rates of cryopreserved mouse embryos.

• 대한치과보철학회지
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• 제40권5호
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• pp.451-469
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• 2002

The purpose of this study was to estimate the effect of a bleaching agent on tooth surfaces and to evaluate the resin bond strength according to different surface treatments on bleached teeth. To prepare for the experimental samples, first, extracted human third molars were used and the body portions of the crowns were cut into four equal-sized specimens. Next, each specimen was mounted in an plastic bottle with self-cured resin and highly polished to have them reveal the enamel or dentin surfaces. Then, the enamel(E) and dentin(D) specimens were divided into four ; 1) non-bleached, laser-treated(NBLA) group 2) bleached, laser-treated(BLLA) group 3) non-bleached, acid-treated(NBAC) group and 4) bleached, acid-treated(BLAC) group. Here, $opalescence^{(R)}$ (10% carbamide peroxide) was used for bleaching agent. The treated specimens were observed by confocal laser scanning microscopy and bonded with composite resin for shear bond test. The following results were obtained from this experiment : 1. Compared with the ENB group, the EBL group was shown be dyed about $20{\mu}m$ deeper with rhodamine B. The DBL group appeared to be caved in at the entry part of the dentinal tubules, was dyed about $20{\mu}m$ deeper and $5{\mu}m$ wider in diameter, compared with the DNB group. 2. In comparison with the EBLAC group, the ENBAC group looked evenly bonded with the resin, while the DNBAC group, compared to DBLAC group, was observed to have its resin tags penetrated about $50{\mu}m$ deeper. Other than those, however, no observable differences between ENBLA and EBLLA group or between DNBLA and DBLLA group were found. 3, At the shear bond test, the ENBAC group was shown to have statistically significant higher shear bond strength than the EBLAC group(p<0.05). No statistically significant differences between the ENBLA and the EBLLA groups were observed(p>0.05). 4. At the shear bond test, the DNBAC group was shown to have statistically significant higher shear bond strength than the DBLAC group(p<0.05). No statistically significant differences between the DNBLA and the DBLLA groups were observed(p>0.05). The in vitro observations above suggest that tooth-bleaching procedure brings a certain change on enamel and dentin surfaces and it weakens the shear bond strength with composite resin when the bleached tooth was acid-treated.

• 대한소아치과학회지
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• 제44권1호
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• pp.20-27
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• 2017

본 연구는 포졸란 시멘트를 기반으로 하는 근관전색제인 Endoseal MTA (Maruchi, Wonju, Korea)를 근관전색제와 근관충전제로 사용하였을 때 나타나는 치근단부 미세누출을 기존의 근관전색제인 AH $Plus^{(R)}$ (Dentsply DeTrey, Konstanz, Germany)와 비교하여 평가하고자 하였다. 만곡되지 않은 영구 전치를 3개의 실험군(A군, E1군, E2군)으로 나누고 A군은 AH $Plus^{(R)}$와 gutta-percha (GP), E1군 은 Endoseal MTA와 GP, E2군은 Endoseal MTA 단독으로 충전하였다. 충전이 완료된 치아는 완전한 경화를 위해 24시간동안 멸균 증류수에 보관하였고 염색액 누출을 위해 치근단 부위를 0.2% 로다민 B 염색 용액에 24시간 동안 노출시켰다. 입체 현미경을 이용하여 미세누출 정도를 평가하였으며 3개의 실험군 사이에 유의한 차이가 관찰되지 않았다. 결론적으로 Endoseal MTA는 GP의 사용에 관계 없이 AH $Plus^{(R)}$와 유사한 밀폐력을 보였으며 이러한 결과는 Endoseal MTA의 근관전색제 및 근관 충전제로써 유용한 사용에 대한 가능성을 제시한다.

• 한국수자원학회:학술대회논문집
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• 한국수자원학회 2005년도 학술발표회 논문집
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• pp.1011-1015
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• 2005

본 실험에서는 부력 조건이 달라질 때 단공방류구에서 정체수역으로 수평방류되는 부력제트의 거동을 규명하였다. LIF (Laser Induced Fluorescence) 시스템을 이용하여 수행하였는데, LIF 시스템은 고해상도의 이미지를 취득할 수 있어 데이터의 정확도가 높으며, 동시에 한 평면상의 농도장을 일시에 측정할 수 있는 장점이 있는 기술이다. LIF 시스템은 크게 세부분으로 구성되어 있는데 방출시스템, 포착시스템, 처리시스템이 그것이다. 실험 조건을 고려해서 온수를 이용하여 주변수와의 밀도차를 재현하였으며, LIF 시스템의 추적입자로 형광염료 Rhodamine B를 사용하였다. 또한, 실험 데이터 취득과정에서 필요한 검정과정을 수행하였는데, LIF 시스템에서 검정과정은 레이저 입사광의 강도가 불균등한 분포를 가지는 점과 주변수의 매질에 의한 근의 감쇠가 발생하는 문제를 해결하기 위한 것이다. LIF 시스템은 부력제트의 농도장을 매우 정밀하게 측정할 수 있는데, 방류밀도 Froude 수가 변함에 따라 측정된 순간이미지를 통해 제트의 진화과정을 상세하고 가시적으로 확인할 수 있었다. 검정과정을 거친 농도 종단면에서 중심선의 연장선이 LIF 시스템에 의해 측정된 순간이미지의 중심선 궤적과 거의 일치하는 것도 알 수 있었다. 또한 LIF 시스템을 통해서 취득된 단일수평부력제트의 궤적과 중심선 희석률을 기존의 상용모형인 VISJET과 CORMIX1에 의해 예측된 결과와 비교$\cdot$분석한 결과, 제트 중심선 궤적의 경우, LIF 시스템을 이용한 측정값은 대체로 VISJET 모형의 결과와 일치하는 것으로 밝혀졌다. 중심선 희석률의 경우, LIF 측정값은 대체로 CORMIX1 모형, Cederwall(1968)의 경험식과 일치하는 경향을 보였다.0\%$일 때가 밸브를 $60\%$와 $80\%$ 개폐시켰을 때보다 $0.3kg/cm^2,\;0.29kg/cm^2$ 낮게 나타나 밸브를 전체 개방 했을 때 관로내의 수압이 상수설계기준에 적합한 수압을 유지함을 알 수 있다. 상수관로 설계 기준에서는 관로내 수압을 $1.5\~4.0kg/cm^2$으로 나타내고 있는데 $6kg/cm^2$보다 과수압을 나타내는 경우가 $100\%$로 밸브를 개방하였을 때보다 $60\%,\;80\%$ 개방하였을 때가 더 빈번히 발생하고 있으므로 대상지역의 밸브 개폐는 $100\%$ 개방하는 것이 선계기준에 적합한 것으로 나타났다. 밸브 개폐에 따른 수압 변화를 모의한 결과 밸브 개폐도를 적절히 유지하여 필요수량의 확보 및 누수방지대책에 활용할 수 있을 것으로 판단된다.8R(mm)(r^2=0.84)$로 지수적으로 증가하는 경향을 나타내었다. 유거수량은 토성별로 양토를 1.0으로 기준할 때 사양토가 0.86으로 가장 작았고, 식양토 1.09, 식토 1.15로 평가되어 침투수에 비해 토성별 차이가 크게 나타났다. 이는 토성이 세립질일 수록 유거수의 저항이 작기 때문으로 생각된다. 경사에 따라서는 경사도가 증가할수록 증가하였으며 $10\% 경사일 때를 기준으로 $Ro(mm)=Ro_{10}{\times}0.797{\times}e^{-0.021s(\%)}$로 나타났다.천성 승모판 폐쇄 부전등을 초래하는 심각한 선천성 심질환이다. 그러나 진단 즉시 직접 좌관상동맥-대동맥 이식술로 수술적 교정을 해줌으로써 좋은 성적을 기대할 수 있음을 보여주었다.특히

• 한국환경과학회지
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• 제20권10호
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• pp.1273-1284
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• 2011

The aim of this research was to evaluate the performance of insoluble electrode for the purpose of degradation of Rhodamine B (RhB) and oxidants generation [N,N-Dimethyl-4-nitrosoaniline (RNO, indicator of OH radical), $O_3$, $H_2O_2$, free Cl, $ClO_2$)]. Methods: Four kinds of electrodes were used for comparison: DSA (dimensional stable anode; Pt and JP202 electrode), Pb and boron doping diamond (BDD) electrode. The effect of applied current (0.5~2.5 A), electrolyte type (NaCl, KCl and $Na_2SO_4$) and electrolyte concentration (0.5~3.5 g/L) on the RNO degradation were evaluated. Experimental results showed that the order of RhB removal efficiency lie in: JP202 > Pb > BDD ${\fallingdotseq}$ > Pt. However, when concerned the electric power on maintaining current of 1 A during electrolysis reaction, the order of RhB removal efficiency was changed: JP202 > Pt ${\fallingdotseq}$ Pb > BDD. The total generated oxidants ($H_2O_2$, $O_3$, free Cl, $ClO_2$) concentration of 4 electrodes was Pt (6.04 mg/W) > JP202 (4.81 mg/W) > Pb (3.61 mg/W) > BDD (1.54 mg/W), respectively. JP202 electrode was the best electrode among 4 electrodes from the point of view of performance and energy consumption. Regardless of the type of electrode, RNO removal of NaCl and KCl (chlorine type electrolyte) were higher than that of the $Na_2SO_4$ (sulfuric type electrolyte) RNO removal. Except BDD electrode, RhB degradation and creation tendency of oxidants such as $H_2O_2$, $O_3$, free Cl and $ClO_2$, found that do not match. RNO degradation tendency were considered a simple way to decide the method which is simple it will be able to determinate the electrode where the organic matter decomposition performance is superior. As the added NaCl concentration was increases, the of hydrogen peroxide and ozone concentration increases, and this was thought to increase the quantity of OH radical.

• Journal of Microbiology and Biotechnology
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• 제25권11호
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• pp.1880-1893
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• 2015

In this study, Pseudomonas R0-14, which was isolated from Arctic soil samples, showed a clear halo when grown on M9 medium agarose plates containing olive oil-rhodamine B as substrate, suggesting that it expressed putative lipase(s). A putative lipase gene, lipR, was cloned from R0-14 by genome walking and Touchdown PCR. lipR encodes a 562-amino-acid polypeptide showing a typical α/β hydrolase structure with a catalytic triad consisting of Ser₁₅₃-Asp₂₀₂-His₂₆₀ and one α-helical lid (residues 103-113). A phylogenetic analysis revealed that LipR belongs to the lipase subfamily I.3. LipR was successfully expressed in Escherichia coli, purified, and biochemically characterized. Recombinant LipR exhibited its maximum activity towards p-nitrophenyl butyrate at pH 8.5 and 60℃ with a K_m of 0.37 mM and a k_cat of 6.42 s^-1. It retained over 90% of its original activity after incubation at 50℃ for 12 h. In addition, LipR was activated by Ca²⁺, Mg²⁺, Ba²⁺, and Sr²⁺, while strongly inhibited by Cu²⁺, Zn²⁺, Mn²⁺, and ethylenediaminetetraacetic acid. Moreover, it showed a certain tolerance to organic solvents, including acetonitrile, isopropanol, acetone, methanol, and tert-butanol. When algal oil was hydrolyzed by LipR for 24 h, there was an enrichment of n-3 long-chain polyunsaturated fatty acids, including eicosapentaenoic acid (1.22%, 1.65-fold), docosapentaenoic acid (21.24%, 2.04-fold), and docosahexaenoic acid (36.98%, 1.33-fold), and even a certain amount of diacylglycerols was also produced. As a result, LipR has great prospect in industrial applications, especially in food and/or cosmetics applications.

• 대한의용생체공학회:의공학회지
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• 제38권1호
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• pp.43-48
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• 2017

우리가 예상했던 DPNs의 지름은 약 500 nm였으며 이는 SEM과 AFM 영상, Size Distribution을 통해 기대했던 것과 유사한 크기를 가진다는 것을 확인하였다. 또한, Zeta potential은 약 $-17.8{\pm}4.4mV$으로 측정되었다. Zeta potential이 +30 mV이상이면 강한 양성을 띤다고 한다. 나노 입자의 Zeta potential이 강한 양성이면 nonspecific cellular interaction이 높아지지만 간에 의해 쉽게 제거되며, hemolytic activity가 높아지기 때문에 약물 전달을 하기에 적합하지 않은 것으로 알려져 있다. 또한 강한 음성이어도 간에 의해 제거될 확률이 높아진다. 하지만 나노 입자의 Zeta potential이 중성이거나 약한 전하를 띠면 혈액에서 제거가 잘 되지 않아 혈액에 오랫동안 남을 수 있어 약물전달에 유리하고, 약 -15 mV의 전하를 띤 입자는 tumor site에 high accumulation됨이 알려져 있다[14]. DPNs의 경우 $-17.8{\pm}4.4mV$이므로 인체에 적용하기에 적합한 것으로 판단된다. DPNs의 Encapsulation Efficiency는 약 $43.8{\pm}6.6%$로 Nano-precipitation과 같은 Bottom-up 방식보다 낮은 수치를 나타내었지만, 독성이 강한 Salinomycin을 사용함으로써 이를 해결할 수 있을 것으로 생각되며 적은 양의 약물만으로 항암효과를 나타낼 수 있을 것으로 기대된다. 암세포와 함께 배양했을 때 형광 현미경으로 확인해본 결과 암세포 주변에 나노 입자가 이동한 것으로 보아 Targeting ligand나 Peptide, Aptamer를 이용하면 더욱 정확한 암세포 표적화를 이룰 수 있을 것으로 예상된다[15]. DPNs의 Drug Carrier로서의 평가는 Loading Amount와 Drug Releasing Profile을 통해 추가로 검증을 할 예정이며, Cell viability를 실행하여 DPNs의 In vitro 항암 효과를 확인하고 In vivo 실험을 진행할 예정이다.

• Journal of Korean Neurosurgical Society
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• 제29권10호
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• pp.1289-1295
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• 2000

Purpose : Recent evidence suggests a possible role for leukocytes in brain injury following ischemia and reperfusion. This study examined the temporal profile of ischemic tissue damage and leukocyte response after transient middle cerebral artery occlusion(MCAO) with reperfusion in the mouse. Methods : Focal cerebral ischemia was made by temporary occluding of the stem of the proximal MCA. Two groups of the mouse were investigated : (1) sham operation(n＝10), and (2)those having the arterial occlusion released after 90 minute(n＝20). By 4 hours(n＝10) and 24 hours(n＝10) after the onset of ischemia-reperfusion, fluorescein videoimages were under-taken in the pial venules of the mouse using a closed cranial window technique. Rhodamine 6G was administered as a $80-100{\mu}l/min$ i.v. loading dose and a $30-40{\mu}l/min$ i.v. maintenance dose in saline to selectively label circulating leukocytes. Neuropathologic evaluation for brain injury was accomplished using the histochemical stain 2,3,5-triphen-yltetrazolium chloride(TTC) and hematoxylin and eosin(H & E) stain. Results : The mean number of adherent leukocytes to cerebral venules in the 90 minutes MCAO and 24 hours reperfusion group were $306{\pm}24$ compared with $72{\pm}8$ in the sham operation group. In the TTC staining method, the cortical infarct affecting 34.8% of hemispheric volume were created in all of animals (n＝10) undergoing 90 minute MCAO with 24 hours reperfusion, but the infarcted area were not found in the other(sham operation and 90 minute MCAO with 4 hours reperfusion)groups. In the H & E stain, the brain tissue following 90 minute MCAO with 4 hours reperfusion revealed only a pyknosis of the nuclei with shrunken cytoplasm, but infiltrated leukocytes were not observed. After 24 hours of reperfusion, a many leukocytes were infiltrated within parenchyma and blood vessles. Conclusions : These findings demonstrate the feasiblity of continous in vivo monitoring of leukocyte adherence in cerebral venules and suggest that reperfusion induced leukocyte adherence to venular endothelium may contribute to tissue injury following focal cerebral ischemia.

• 공업화학
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• 제28권6호
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• pp.705-713
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• 2017

독창적 물질인 $Bi_2WO_6-GO-TiO_2$ 나노복합체를 쉬운 수열법에 의해 성공적으로 합성하였다. 수열반응을 하는 동안, 그래핀 시트 위에 $Bi_2WO_6$와 $TiO_2$를 도포하였다. 합성한 $Bi_2WO_6-GO-TiO_2$ 복합체형 광촉매는 X-선 회절법(XRD), 주사전자현미경(SEM), 에너지 분산 X-선(EDX) 분석, 투과전자현미경(TEM), 라만분광법, UV-Vis 확산반사 분광법(UV-vis-DRS), 및 X-선 광전자분광기(XPS)에 의하여 특성화하였다. $Bi_2WO_6$ 나노입자는 불규칙한 dark-square block 나노 플페이트 형상을 보였으며, 이산화티탄 나노입자는 퀜텀 도트 사이즈로 그래핀 시트 위 표면을 덮고 있었다. 로다민 비의 분해는 농도감소의 측정과 함께 UV 분광법에 의하여 관찰하였다. 합성된 물질의 광촉매 반응은 Langmuir-Hinshelwood 모델과 띠 이론으로 설명하였다.