• Journal of Microbiology and Biotechnology
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• 제22권12호
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• pp.1782-1789
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• 2012

We have developed a novel type of polymer nanogel loaded with anticancer drug based on bio-derived poly(${\gamma}$-glutamic acid) (${\gamma}$-PGA). ${\gamma}$-PGA is a highly anionic polymer that is synthesized naturally by microbial species, most prominently in various bacilli, and has been shown to have excellent biocompatibility. Thiolated ${\gamma}$-PGA was synthesized by covalent coupling between the carboxyl groups of ${\gamma}$-PGA and the primary amine group of cysteamine. Doxorubicin (Dox)-loaded ${\gamma}$-PGA nanogels were fabricated using the following steps: (1) an ionic nanocomplex was formed between thiolated ${\gamma}$-PGA as the negative charge component, and Dox as the positive charge component; (2) addition of poly(ethylene glycol) (PEG) induced hydrogen-bond interactions between thiol groups of thiolated ${\gamma}$-PGA and hydroxyl groups of PEG, resulting in the nanocomplex; and (3) disulfide crosslinked ${\gamma}$-PGA nanogels were fabricated by ultrasonication. The average size and surface charge of Dox-loaded disulfide cross-linked ${\gamma}$-PGA nanogels in aqueous solution were $136.3{\pm}37.6$ nm and $-32.5{\pm}5.3$ mV, respectively. The loading amount of Dox was approximately 38.7 ${\mu}g$ per mg of ${\gamma}$-PGA nanogel. The Dox-loaded disulfide cross-linked ${\gamma}$-PGA nanogels showed controlled drug release behavior in the presence of reducing agents, glutathione (GSH) (1-10 mM). Through fluorescence microscopy and FACS, the cellular uptake of ${\gamma}$-PGA nanogels into breast cancer cells (MCF-7) was analyzed. The cytotoxic effect was evaluated using the MTT assay and was determined to be dependent on both the concentration and treatment time of ${\gamma}$-PGA nanogels. The bio-derived ${\gamma}$-PGA nanogels are expected to be a well-designed delivery carrier for controlled drug delivery applications.

• 한국안광학회지
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• 제20권4호
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• pp.437-443
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• 2015

목적: 본 연구는 3-chloropyridine-4-carboxylic acid와 3-fluoropyridine-4-carboxylic acid를 첨가제로 사용하여 소프트 콘택트렌즈 제조 후 물리적 및 광학성 특성을 분석하여 콘택트렌즈 재료로서 활용도를 알아보았다. 방법: 본 실험에서는 소프트 하이드로젤 콘택트렌즈의 주재료인 HEMA (2-hydroxyethyl methacrylate)와 AA(acylic acid), MMA(methacrylic acid) 그리고 개시제인 AIBN(azobisisobutyronitrile), 교차결합제인 EGDMA(ethylene glycol dimethacrylate)를 사용하여 공중합 하였다. 결과: 생성된 고분자의 물리적 특성을 측정한 결과, 굴절률 1.4320~1.4342, 함수율 34.54~37.15%, 접촉각 $57.82{\sim}79.57^{\circ}$, 인장강도 0.2872~0.3608로 각각 나타났고 광학적 특성 측정 결과 가시광선 영역에서는 투과율이 88.8~90.6%로 나타났으며, UV-B 76.8~82.4%, UV-A 84.6~86.6%로 각각 나타났다. 결론: 본 실험 결과 3-chloropyridine-4-carboxylic acid 및 3-fluoropyridine-4-carboxylic acid를 첨가제로 사용하였을 때 고습윤성 및 자외선 차단기능을 가진 콘택트렌즈 재료로 활용될 수 있을 것으로 판단된다.

• 생체재료학회지
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• 제22권4호
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• pp.328-336
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• 2018

Background: Biogenic fabrication of silver nanoparticles from naturally occurring biomaterials provides an alternative, eco-friendly and cost-effective means of obtaining nanoparticles. It is a favourite pursuit of all scientists and has gained popularity because it prevents the environment from pollution. Our main objective to take up this project is to fabricate silver nanoparticles from lichen, Usnea longissima and explore their properties. In the present study, we report a benign method of biosynthesis of silver nanoparticles from aqueous-ethanolic extract of Usnea longissima and their characterization by ultraviolet-visible (UV-vis), Fourier transform infrared (FTIR) spectroscopy, transmission electron microscopy (TEM) and scanning electron microscopy (SEM) analyses. Silver nanoparticles thus obtained were tested for antimicrobial activity against gram positive bacteria and gram negative bacteria. Results: Formation of silver nanoparticles was confirmed by the appearance of an absorption band at 400 nm in the UV-vis spectrum of the colloidal solution containing both the nanoparticles and U. longissima extract. Poly(ethylene glycol) coated silver nanoparticles showed additional absorption peaks at 424 and 450 nm. FTIR spectrum showed the involvement of amines, usnic acids, phenols, aldehydes and ketones in the reduction of silver ions to silver nanoparticles. Morphological studies showed three types of nanoparticles with an abundance of spherical shaped silver nanoparticles of 9.40-11.23 nm. Their average hydrodynamic diameter is 437.1 nm. Results of in vitro antibacterial activity of silver nanoparticles against Staphylococcus aureus, Streptococcus mutans, Streptococcus pyrogenes, Streptococcus viridans, Corynebacterium xerosis, Corynebacterium diphtheriae (gram positive bacteria) and Escherichia coli, Klebsiella pneuomoniae and Pseudomonas aeruginosa (gram negative bacteria) showed that it was effective against tested bacterial strains. However, S. mutans, C. diphtheriae and P. aeruginosa were resistant to silver nanoparticles. Conclusion: Lichens are rarely exploited for the fabrication of silver nanoparticles. In the present work the lichen acts as reducing as well as capping agent. They can therefore, be used to synthesize metal nanoparticles and their size may be controlled by monitoring the concentration of extract and metal ions. Since they are antibacterial they may be used for the treatment of bacterial infections in man and animal. They can also be used in purification of water, in soaps and medicine. Their sustained release may be achieved by coating them with a suitable polymer. Silver nanoparticles fabricated from edible U. longissima are free from toxic chemicals and therefore they can be safely used in medicine and medical devices. These silver nanoparticles were stable for weeks therefore they can be stored for longer duration of time without decomposition.

• Clinical and Experimental Reproductive Medicine
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• 제45권3호
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• pp.143-148
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• 2018

Objective: The favored method of preserving fertility in young female cancer survivors is cryopreservation and autotransplantation of ovarian tissue. Reducing hypoxia until angiogenesis takes place is essential for the survival of transplanted ovarian tissue. The aim of this study was to investigate the role of angiopoietin-1 (Angpt-1), angiopoietin-2 (Angpt-2), and vascular endothelial growth factor (VEGF) in ovarian tissue grafts that were cryopreserved using two methods. Methods: Ovarian tissues harvested from ICR mice were divided into three groups: group I (control), no cryopreservation; group II, vitrification in EFS (ethylene-glycol, ficoll, and sucrose solution)-40; and group III, slow freezing in dimethyl sulfoxide. We extracted mRNA for VEGF, Angpt-1, and Angpt-2 from ovarian tissue 1 week following cryopreservation and again 2 weeks after autotransplantation. We used reverse transcriptase-polymerase chain reaction to quantify the levels of VEGF, Angpt-1, and Angpt-2 in the tissue. Results: Angpt-1 and Angpt-2 expression decreased after cryopreservation in groups II and III. After autotransplantation, Angpt-1 and Angpt-2 expression in ovarian tissue showed different trends. Angpt-1 expression in groups II and III was lower than in group I, but Angpt-2 in groups II and III showed no significant difference from group I. The vitrified ovarian tissues had higher expression of VEGF and Angpt-2 than the slow-frozen ovarian tissues, but the difference was not statistically significant. Conclusion: Our results indicate that Angpt-2 may play an important role in ovarian tissue transplantation after cryopreservation although further studies are needed to understand its exact function.

• 한국패류학회지
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• 제29권3호
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• pp.251-258
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• 2013

본 연구에서는 굴, Crassostrea gigas의 계획적인 인공종묘생산 및 우량형질의 정자를 보존하기 위하여 냉동보존 시 적정 결빙억제제 (cryprotective agent, CPA) 및 농도를 알아보고자, CPA 종류 및 농도별 독성 및 냉동보존 효과를 파악하고 해동 후 정자의 세포 손상을 조사하였다. CPA 종류 및 농도별 침지시간에 따른 굴 정자의 독성평가를 실시한 결과, 생존율 및 운동성은 DMSO가 가장 좋았으며, 다음으로 EG, glycerol, Methanol 순이었다. 희석액으로 여과해수를 사용하여 CPA 종류 및 농도별 냉동보존 결과, 15% DMSO로 냉동보존 한 정자의 생존율 및 운동성이 가장 높았다. 굴의 냉동/해동 정자를 주사전자현미경으로 관찰한 결과, DMSO, EG, methanol, glycerol 순으로 세포 손상이 적었으며, 농도는 15, 20, 10, 5% 순이었다. 이상의 결과를 종합한 결과, 굴 정자의 냉동보존 시 여과해수를 희석액으로 사용할 경우 적정 CPA는 DMSO이며, 농도는 15%였다.

• 대한임상독성학회지
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• 제17권2호
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• pp.47-57
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• 2019

Purpose: Osmolar gap (OG) has been used for decades to screen for toxic alcohol levels. However, its reliability may vary due to several reasons. We validated the estimated ethanol concentration formula for patients with suspected poisoning and who visited the emergency department. We examined discrepancies in the ethanol level and patient characteristics by applying this formula when it was used to screen for intoxication due to toxic levels of alcohol. Methods: We retrospectively reviewed 153 emergency department cases to determine the measured levels of toxic ethanol ingestion and we calculated alcohol ingestion using a formula based on serum osmolality. Those patients who were subjected to simultaneous measurements of osmolality, sodium, urea, glucose, and ethanol were included in this study. Patients with exposure to other toxic alcohols (methanol, ethylene glycol, or isopropanol) or poisons that affect osmolality were excluded. OG (the measured-calculated serum osmolality) was used to determine the calculated ethanol concentration. Results: Among the 153 included cases, 114 had normal OGs (OG≤14 mOsm/kg), and 39 cases had elevated OGs (OG>14). The mean difference between the measured and estimated (calculated ethanol using OG) ethanol concentration was -9.8 mg/dL. The 95% limits of agreement were -121.1 and 101.5 mg/dL, and the correlation coefficient R was 0.7037. For the four subgroups stratified by comorbidities and poisoning, the correlation coefficients R were 0.692, 0.588, 0.835, and 0.412, respectively, and the mean differences in measurement between the measured and calculated ethanol levels were -2.4 mg/dL, -48.8 mg/dL, 9.4 mg/dL, and -4.7 mg/dL, respectively. The equation plots had wide limits of agreement. Conclusion: We found that there were some discrepancies between OGs and the calculated ethanol concentrations. Addition of a correction factor for unmeasured osmoles to the equation of the calculated serum osmolality would help mitigate these discrepancies.

• 광물과 암석
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• 제35권4호
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• pp.431-445
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• 2022

점토질 암석의 광물함량을 효과적이고 실용적으로 분석하는 방법을 X-선 분말회절분석 실험을 통하여 연구하였다. 점토질 암석의 X-선 분말회절 정량분석을 위해서는 측면마운팅(side mounting) 방법에 의한 무작위 배향(random orientation)의 전암(whole-rock) 분말시료의 준비가 필요하다. 또한, 암석을 구성하고 있는 점토광물의 감정을 위하여 2 ㎛ 이하 점토입도의 배향성 마운트(oriented mount)시편의 준비와 에틸렌글리콜 처리, 열처리 등의 실험과정이 요구된다. 정량분석을 위하여 RIR(reference intensity ratio)방법과 리트벨트(Rietveld) 회절도 계산 방법을 사용하였다. RIR값을 사용하여 전암 X-선 회절도로부터 총 점토 함량과 비점토광물(non-clay minerals)들의 함량을 얻을 수가 있었다. 또한, 점토입도의 배향성 X-선 회절도로부터는 각각 점토광물의 상대함량을 계산하여 이를 총 점토광물에 할당할 수가 있었다. 전암 X-선 회절의 리트벨트 방법에서는 10°(2θ) 미만의 X-선 회절 영역은 제외한 후에 리트벨트 회절도를 계산하였을 때 효과적인 정량분석 값을 얻을 수 있었다. 분석결과는 RIR방법과 리트벨트 방법이 서로 근사한 정량분석 값을 보여주었다. 따라서, 연구결과는 실험실에서의 일상적인 점토질암의 광물정량분석을 성공적으로 수행하는 것이 가능함을 지시한다. 그러나, 점토광물은 화학적 및 구조적 특정이 다른 수많은 변종이 존재하기 때문에 점토질암의 정량분석은 아직도 도전해야 하는 과제이다.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.76-76
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• 2001

The objective of this study was to optimize the vitrification method of in vitro produced bovine embryos. Thus, in vitro produced embryos at 8 cell, morula and blastocyst stages were vitrified on electron microscope grids (EM grids) or in open pulled straws (OPS) with EG5.5 (5.5 M ethylene glycol, 1.0 M sucrose and 10% FBS in m-DPBS medium) freezing solution and their survival rates after thawing were compared. The embryos on EM grids or in OPS were briefly exposed to EG5.5 freezing solution and plunged directly into liquid nitrogen within 30 to 35 sec. Post-thawed embryos were serially diluted in 0.5, 0.25 and 0.125 M sucrose in m-DPBS, each for 1 min, and then cultured in CRI aa medium supplemented with 10% FBS. Embryonic survival rate was assessed as re-expanded and hatched rates of those embryos after warming. The rates of re-expansion embryos did not significantly different between EM grid (8 cell: 42.10%, morula: 66.66% and blastocyst: 77.08%) and OPS (8 cell: 47.36%, morula: 61.90% and blastocyst: 83.33%) methods. In addition, the hatched rates in EM grid (8 cell: 31.57%, morula: 57.14% and blastocyst: 72.91%) were similar to those in OPS (8 cell. 34.21%, morula: 50.00% and Blastocyst: 77.08%). Interestingly, even at the same blastocyst stage, the in vitro survival of day 7 embryos (EM grid: 79.48 and OPS: 87.18%) was higher than those of day 8 embryos (EM grid: 72.10 and OPS: 82.06%). The total cell number of blastocyst developed in vitro after vitrification was examined with Hoechst 33342 staining to compare the embryo quality among different treatment groups. The total cell number of blastocyst was not significantly different between vitrified groups (EM grid: 162.4$\pm$8.0 and OPS: 158.4$\pm$7.1) and unvitrified control (168.0$\pm$5.6). These results indicate that both vitrification containers can provide the high rate of embryo survival. Moreover, the OPS container may not need a cap to protect the container from floating after immersion in L$N_2$. Therefore, this study suggest that bovine embryos can be cryopreserved easily, effectively and successfully by vitrification method using EM grid or OPS with EG5.5 freezing solution. In the future, the Pregnancy rate would be investigated after transfer of our vitrified embryos into the appropriated recipients.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2002년도 국제심포지엄
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• pp.78-78
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• 2002

The role of heat shock proteins in shielding organism from environmental stress is illustrated by the large-scale synthesis of these protein by the organism studied to date. However, recent evidence also suggests an important role for heat shock protein in fertilization and early development of mammalian embryos. Effects of elevated in vitro temperature on in vitro produced bovine embryos were analysed in order to determine its impact on the expression of heat shock protein 70 (HSP70) by control and frozen-thawed after in vitro fertilization (IVF) or nuclear transfer (NT). The objective of this study was to assess the developmental potential in vitro produced embryos with using of the various containers and examined expression and localization of heat shock protein 70 after it's frozen -thawed. For the vitrification, in vitro produced embryos at 2 cell, 8 cell and blastocysts stage after IVF and NT were exposed the ethylene glycol 5.5 M freezing solution (EG 5.5) for 30 sec, loaded on each containers such EM grid, straw and cryo-loop and then immediately plunged into liquid nitrogen. Thawed embryos were serially diluted in sucrose solution, each for 1 min, and cultured in CRI-aa medium. Survival rates of the vitrification production were assessed by re-expanded, hatched blastocysts. There were no differences in the survival rates of IVF using EM grid, cryo-loop. However, survival rates by straw were relatively lower than other containers. Only, nuclear transferred embryos survived by using cryo-loop. After IVF or NT, in vitro matured bovine embryos 2 cell, 8 cell and blastocysts subjected to control and thawed conditions were analysed by semiquantitive reverse transcription polymerase chain reaction methods for hsp 70 mRNA expression. Results revealed the expression of hsp 70 mRNA were higher thawed embryos than control embryos. Immunocytochemistry used to localization the hsp70 protein in embryos. Two, 8-cell embryos derived under control condition was evenly distributed in the cytoplasm but appeared as aggregates in some embryos exposed frozen-thawed. However, under control condition, blastocysts displayed aggregate signal while Hsp70 in frozen-thawed blastocysts appeared to be more uniform in distribution.

• 한국가금학회지
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• 제41권4호
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• pp.261-270
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• 2014

동결 닭 PGCs의 생식계열 키메라를 이용한 생체에의 복원을 실용화 하기 위해서는, 닭 PGCs의 동결 보존 기술의 향상에 의해 동결 및 융해 후의 많은 생존 세포를 확보하는 것과, 생식계열 키메라의 제작 효율을 높이는 것이 반드시 필요하다. 닭 PGCs는 배양 5.5~6일령의 닭 원시 생식선으로부터 채취하고, MACS 방법에 의해서 순수 닭 PGCs를 분리했다. 15% 각각의 EG와 DMSO를 동결 보호제로 사용한 처리군이 각 군의 농도에 상관없이 유의적(p<0.05)으로 PG 처리군보다 동결 및 융해 후의 세포의 생존율이 높음을 확인하였다. 특히, 10% EG+FBS 조합의 처리군에서 상업용 닭인 한협육종협회3호종(B : 88.7%)이 오계종(A : 85.1%), 아사 브라운종(C : 84.6%) 그리고 화이트 레그혼종(D : 85.9%)의 세 품종보다 동결 및 융해 후의 닭 PGCs의 생존율이 유의적(p<0.05)으로 높음을 확인하였다. 간이 동결에 있어서 가장 높은 생존율을 보인 10% EG이 10% DMSO와 함께 최적의 동결 보호제로서 사용 가능성을 확인하였다.