• 한국가축번식학회지
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• 제17권2호
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• pp.103-111
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• 1993

These experiments were undertaken to establish the optimal culture systems for in vitro maturation, fertilization and subsequently embryonic development of porcine immature follicular oocytes isolated from the ovary of slaughtered pigs. Porcine ovaries were brought to the laboratory from local slaughter house within 1 hour after slaughtering and cumulus oocytes complexes were recovered from antral follicles (3~5mm) with 23 gauge needle. To maturate follicular oocytes, cumulus oocytes complexes were washed three times with TCM-199 containing 25mM HEPES and incubated (39$^{\circ}C$, 5% CO2 in air) for 42hrs. Ejaculated and liquid storaged boar spermatozoa capacitated with different sperm capacitation methods and media were prepared forfertilizaing of matured follicular oocytes in vitro. Fertilization was performed by adding 5~10${mu}ell$ of capacitated spermatozoa containing 1~5$\times$105 sperm/ml to droplets. Eighteen to twenty-eight hours after sperm insemination, fertilized eggs were washed three times with culture media and transferred to the culture media. The fertilization rates of in vitro matured follicular oocytes cultured in B. O., TCM-HEPES, m-KRB, and TALP-II media were 61.3%, 83.0%, 88.9% and 89.2%, respectively. In addition, the polyspermy rates were 60.7%, 66.5%, 53.8%, and 43.9%, respectively. These data indicated that the highest of fertilization and the lowest of polyspermy rate was shown in TALP-II medium. Spermatozoa capacitated by caffeine, heparin, and percoll density gradient treatment in the 4 different media, the fertilization rates were 33.0~57.2%, 39.9~90.2%, and 52.6~92.8%, respectively, showing the lowest rate in caffeine treatment. The development rate of follicular oocytes, fertilized with the spermatozoa capacitated by caffeine, heparin, and percoll gradient in the TALP-II medium, upto 2 to 4-cell stages were 32.6%, 74.5% and 70.9%, respectively. Finally, fertilization rates of follicular oocytes cultured with follicular fluid containing medium from 10 to 100% were 61.2~94.1% and the rates (90~94%) with 10~20% follicular fluids were significantly higher than those (85.3%) of cultured in the media without follicular fluid. In addition, the rates of pronucleus formation were also higher in follicular fluid treated group (73.1~83.0%) than those (64.7%) of oocytes cultured without follicular fluid. The highest fertilization and pronucleus formation rates was found in oocytes cultured with 10% follicular fluid. These results suggest that the addition of heparin or percoll density gradient method is better capacitation method. Furthermore, the addition of porcine follicular fluid to the fertilization medium may improve the fertilization rates and formation of pronucleus.

• The Korean Journal of Physiology
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• 제24권1호
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• pp.161-170
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• 1990

The effects of $H^{+}$ on the arterial contraction and their mechanisms were investigated in the renal artery of a rabbit. The helical strips of isolated renal artery were immersed in the HEPES-buffered or $CO_{2}/HCO_{3}^{-}$-buffered Tyrode's solution. The contractions induced by agonists (norepinephrine, histamine, serotonin and angiotensin II) or high $K^{+}$ were observed with change of extracellular or intracellular $H^{+}$ concentration. The contractions induced by norepinephrine, histamine, serotonin, angiotensin II or high $K^{+}$ in HEPES-buffered Tyrode's solution were inhibited by increase in extracellular $H^{+}$ concentration and potentiated by decrease in extracellular $H^{+}$ concentration. The degrees of these effects were most evident in the contraction induced by serotonin and angiotensin II, moderate in those by histamine and high $K^{+}$, and least in those by norepinephrine. Maximal contraction by norepinephrine, histamine and high $K^{+}$ were not influenced by change in extracellular $H^{+}$ concentration, but influenced in those contration by serotonin and angiotensin II. The attenuated contractions by an acidic pH were not returned to the level of contraction at normal pH (7.4) by elevation of extracellular $Ca{2+}$ concentration. The agonists (norepinephrine, histamine and serotonin)-induced contractions in $Ca{2+}$-free Tyrode's solution were also attenuated by increase in extracellular $H^{+}$ concentration and potentiated by decrease in extracellular $H^{+}$ concentration. Elevation of $Pco_{2}$ in the $CO_{2}/HCO_{3}^{-}$-buffered Tyrode's solution, which increase the intracellular $H^{+}$ concentration, at constant extracellular pH (7.4), increased the contraction by 30 mM $K^{+}$. From the above results, it is suggested that the decrease in contractions by increase in extracellular $H^{+}$ concentration may be resulted from that $H^{+}$ make the receptors less sensitive to agonists and cell membrane hyperpolarize and then inhibit the $Ca{2+}$ influx as well as $Ca{2+}$ release from intracellular $Ca{2+}$ storage site.

• 한국발생생물학회지:발생과생식
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• 제3권1호
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• pp.39-44
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• 1999

성선자극호르몬으로 자극된 난소로부터 회수된 사람의 성숙난자의 전핵형성과 초기발생에 정자의 운동성과 성숙도가 미치는 영향을 조사하였다. 세포질내정자주입 (ICSI)은 HEPES-buffer mTCM-199 배양액에서 실시하였다. 체내에서 성숙된 난구세포부착난자를 ICS에 의하여 사정된 운동성 정자 또는 비운동성 정자로 수정하였을 때 운동성 정자로 수정된 난자가 비운동성 정자로 수정된 난자보다 전핵형성율이 높았다 (79.8% vs 51.7% ; p<0.002)). 그러나 체내에서 성숙된 난구세포부착난자를 ICS에 의해 정소 내 운동성 정자 또는 비운동성 정자로 수정하였을 때 운동성 정자와 비운동성 정자 사이의 전핵형성율은 차이가 없었다. 10.0 mM lactate, 0.5 mM pyruvate, 0.2 mM taurine, 1.0 mM glutamine, 2.22 mM MEM amino acids, vitamin 그리고 10% 사람 난포액이 포함된 수정 Tyrode 배지에서 전핵이 형성된 수정란의 초기 발생은 정자의 채취원과 운동성에 관계없이 9∼16세포기로 발생하였다.

• 폴리머
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• 제28권6호
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• pp.478-486
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• 2004

소장점막하조직은 콜라젠과 글리코스아미노글리칸 및 세포활성을 촉진하는 성장인자들로 구성되어 있다. 최근에는 이종이식의 면역 거부반응이 없는 생체물질로서 응용되고 있고 SIS에 함유되어 있는 성장인자는 전층 피부손상층을 치료하는데 중요한 역할을 한다고 알려져 있다. 우리는 본래의 SIS 쉬트와 아세트산으로 처리하여 팽윤시킨 SIS 쉬트를 제조하였고, 각각에 대해 1겹과 5겹의 SIS 쉬트를 제조하였다. 이를 SEM을 통해 표면 및 단면의 형태를 확인하였고, 증류수, 인산염 완충액, HBSS (Hank's balanced salt solution)완충액, 트리스 완충액, HEPES (N-[2-hydroxyethyl) piperazine-N'-(2-ethanesulfonic acid]) 완충액에서의 물 흡수성, pH에 따른 물 흡수성 그리고 시간에 따른 물 흡수성을 비교 실험하였다. 본래의 SIS 쉬트보다 산 처리된 SIS 쉬트가, 증류수보다는 완충액에서의 물 흡수성이 높음을 확인하였다. 중성 용액보다 산성과 염기성 용액에서 SIS 쉬트는 팽윤되어 더 많은 물을 흡수하였다. 또한, 증류수와 HEPES 완충액에서 시간에 따른 물 흡수성 실험을 한 결과 1일 이후부터 10일 동안 약 200%물을 지속적으로 흡수하였다. 이로써 SIS 쉬트와 산 처리된 SIS 쉬트가 창상 치료를 위한 드레싱제와 생분해성이식 재료로서 사용가능하리라 판단된다.

• Journal of Chest Surgery
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• 제30권6호
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• pp.553-558
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• 1997

동종동맥판은 심장판막질환, 선천성 심기형 및 대동맥 질환의 치료에 있어서 우수한 판막도관으로 사용 되고 있다. 이 때 동종동맥 판의 장기성적을 좌우하는데 있어서 혈관내피세포의 생육성이 중요한 역활을 할 것이다. 혈관내피세포의 생육성을 평가하기 위하여 현재 임상에서 사용되는 보존방법으로 보존처리된 성돈의 대동맥판 및 대동맥 벽을 collagenase로 분해시켜서 순수한 내퍼세포군을 획득한 뒤, 혈관내피세포에 특이한 친화성을 갖는 GSA-FTTC(Criffonia simplicifolia agglutininfluorescein isothiocyanate)와 반응시켰다. 이 내피세포군을 세척한 다음, 살아있는 세포에는 침착되지 않는 Pl(Ropidium iodide)와 반응시켰다. 이렇게 처리된 내피세포군을 Row Cytometry 로 분석하여 GSA-FTIC(+), Pl(-) 인 세포를 생육성을 유지한 것으로 평가하였다. 동종동맥판은 $4^{\circ}C의$ 멸균용액에 24시간 담궈 멸균처리를 한 후, 2개군으로 나누어 (1군)은 $4^{\circ}C$ RPM 1640 with HEPES buffer cultlue medium with 10% fetal bovine uTm 용액에 1~14일간 보존하였고 (2군)은 냉동보존을 하였다. 조직의 획득과정과 멸균과정에서 각각 22.8%와 24.4%의 생육성이 소\ulcorner되었다. (1군) 에서는 14일의 보존기간 동안 11.9%의 생육성감소가 일어났고 (2군) 에서는 13.7%의 생육성감소가 일어났다. 이 실험의 결과로 동종동맥 판의 보존처리과정 초기에 대부분의 생육성소실이 일어나며, 14일간의 냉장보존이나 냉동보존 후에도 약 40%의 생육성이 보존됨을 알 수 있었다. 또한 혈관내피세포가 판막에서 얻어진 경우나 동맥벽에서 얻어진 경우에서 생육성의 차이는 없었다.

• 대한임상검사과학회지
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• 제56권1호
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• pp.43-51
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• 2024

[⁶⁸Ga]Ga-FAPI-04는 암세포에 과발현 되어 있는 fibroblast activation protein (FAP)에 특이적으로 결합하는 FAP 저해제(FAP inhibitor, FAPI)에 방사성동위원소 ⁶⁸Ga을 표지한 방사성의약품이다. 본 연구에서는 국내에서 제작된 카세트기반 자동합성장치를 사용하여 [⁶⁸Ga]Ga-FAPI-04를 제조하는 방법을 개발하였다. [⁶⁸Ga]Ga-FAPI-04의 합법을 개발하기 위해 완충액 HEPES의 농도, 반응시간, FAPI-04 전구체 양, 반응온도에 따른 표지효율을 확인하였다. [⁶⁸Ga]Ga-FAPI-04는 2 M HEPES를 사용하여 pH 3.85에서 반응할 경우 가장 높은 표지효율을 획득할 수 있었고, 반응시간이 10분일 경우와 25 ㎍의 FAPI-04 전구체를 사용할 경우 및 100℃에서 반응할 경우 가장 높은 표지효율을 획득할 수 있었다. 또한 최종 합성된 [⁶⁸Ga]Ga-FAPI-04는 모든 품질기준을 만족하여 본 연구를 통해 개발된 [⁶⁸Ga]Ga-FAPI-04의 합성법은 FAPI 기반 방사성의 약품생산에 활용도가 높을 것으로 예상된다.

• Journal of Pharmaceutical Investigation
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• 제31권2호
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• pp.101-106
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• 2001

Alginate-chitosan (anion-cationic polymeric complex) was prepared to control the release rate of silver sulfadiazine (AgSD). Na-alginate (2%) solution containing AgSD was gelled in $CaCl_2$ solution. The gel beads formed were immediately encapsulated with chitosan (CS). The gel matrix and membrane were then reinforced with chondroitin-6-sulfate (Ch6S). Release rate of AgSD from the gel matrix was investigated by placing alginate beads in the sac of cellulose membrane simmered in HEPES-buffer solution. The concentration of AgSD released was analyzed by UV at 264 nm. Incorporation capacity of AgSD in Ca-alginate gel was more than 90%. Alginate-Ch6S-CS could control the release rate of AgSD. The amount of AgSD release was dependent on the AgSD loading dose. Incorporation of tripolyphosphate (polyanionic crosslinker) onto the alginate-Ch6S-CS bead increased the release rate of AgSD. Collagen-coating had no influence on the AgSD release rate. Alginate-Ch6S-CS beads with a sufficiently high AgSD encapsulation were capable of controlling the release of the drug over 10 days. In summary, alginate-Ch6S-CS beads could be used as a sustained delivery for AgSD and provide local targeting with low silver toxicity and patient discomfort.

• Biomolecules & Therapeutics
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• 제11권3호
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• pp.174-177
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• 2003

The present experiment was performed to investigate the protective effects of paeonol isolated from Moutan Cortex Radicis on primary cultured rat hepatocytes exposed to Br-A23187 ($Ca^{2+}$ ionophore). Br-A23187 is frequently used as a model of cell killing as inducing both necrotic and apoptotic cell death. Hepatocytes were isolated by collagenase perfusion from livers of fasted male Sprague Dawley rats and cultured overnight. Cell viability was determined by propidium iodide using fluorocytometry in Krebs-Ringer-HEPES buffer at pH 7.4. In addition, intracellular calcium was measured by excitation at 340 and 380 nm and emission at 505 nm using a luminescence spectrophotometer. Paeonol (20-100 ${\mu}M$) inhibited cell killing induced by 10 ${\mu}M$ Br-A23187, in a dose-dependent manner. Paeonol also reduced increased intracellular calcium level when hepatocytes were exposed to Br-A23187. Therefore, the present results suggest that paeonol protects the hepatocytotoxicity induced by Br-A23187, via inhibiting the influx of calcium into into rat hepatocytes.

• 한국지하수토양환경학회:학술대회논문집
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• 한국지하수토양환경학회 2000년도 창립총회 및 춘계학술발표회
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• pp.52-55
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• 2000

The effects of environmental conditions, initial dissolved oxygen concentrations, pH, and the presence of electron carrier vitamin B$_{12}$ , on the reduction rate of TNT by Fe$^{0}$ was Quantitatively analyzed using a batch reactor. In all experiments, TNT reduction was best described with a first order reaction and the reduction rate decreased with the increase in the initial DO concentration. However, the specific reaction rate did not decrease linearly with the increase in the initial DO concentration. In the presence of HEPES buffer 0.2 and 2.0 mM(pH 5.7$\pm$0.2), the specific reaction rate increased more than 5.8 times, which showed reduction rate is rather significantly influenced by the pH of the solution. To test the possibility of reaction rate enhancement, well-known electron carrier(or mediator), vitamin B$_{12}$ has augmented besides Fe$^{0}$ . In the presence of 8.0 $\mu\textrm{g}$/L of vitamin B$_{12}$ , the specific reaction rate increased as much as 14.6 times. The results indicate that the addition of trace amount of vitamin B$_{12}$ can be a promising rate controlling option for the removal of organics using a Fe$^{0}$ filled permeable reactive barrier.

• Bulletin of the Korean Chemical Society
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• 제35권11호
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• pp.3326-3330
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• 2014

A quinoline-thiooxorhodamine conjugate fluorescent sensor (1) has been synthesized. Sensor 1 exhibits high selectivity and sensitivity to $Hg^{2+}$ in $H_2O$/DMSO (95/5, v/v, HEPES 20 mM, pH = 7.4) solution with fluorescence detection. Other tested metal ions do not induce any significant fluorescence intensity changes. Sensor 1 interacts with $Hg^{2+}$ through a 1:1 binding stoichiometry with a good anti-inference ability. In addition, fluorescent imaging of $Hg^{2+}$ in Hela cells is also successfully demonstrated.