• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1998년도 Proceedings of UNESCO-internetwork Cooperative Regional Seminar and Workshop on Bioassay Guided Isolation of Bioactive Substances from Natural Products and Microbial Products
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• pp.192-192
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• 1998

The blood - brain barrier (BBB) expresses high concentrations of transferrin receptor, and it was revealed that anti-transferrin receptor mouse monoclonal antibody (OX26) undergoes transcytosis through the BBB. This property allows the OX26 to serve as a brain drug delivery vector. In an attempt to produce broadly useful targeting agents, genetic engineering and expression techniques have been used to produce antibody-avidin (AV) fusion protein (OX26 IgG3C$\_$H/3-AV). In the present study we estimated the BBB permeability and stability of genetically engineered vector.

• Clinical and Experimental Reproductive Medicine
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• 제17권1호
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• pp.81-85
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• 1990

We cryopreserved mouse morulae by a simple ultra-rapid method of freezing embryos directly in $LN_2$ after holding 2min in a $LN_2$ vapor, and thawed them in $37^{\circ}C$ water bath. The time requirements for permeation and dehydration by 2.0 M glycerol and 0.2 M sucrose before freezing were studied. When the embryos were equilibrated for 10 min, the optimun post-thaw survival was obtained. Embryos those developed normally to blastocyst after in vitro culture for over 24hrs were regarded as survival ones. Two experiments to assess post-thaw survival following predehydration in various mixtures of glycerol and sucrose were also accomplished. When sucrose was held constant (0.2 M) and glycerol concentration varied (1.5-3.5 M), post-thaw survival was best (78.0%) in 3.0 M glycerol. When glycerol was held constant (3.0M) and sucrose concentration varied (0.0-1.0M), optimun post-thaw survival (78.0%) was found in 0.2 M sucrose.

• Parasites, Hosts and Diseases
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• 제37권1호
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• pp.21-26
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• 1999

The growth and development of Gymnophalloides seoi were studied in C3H/HeN mice and effects of immunosuppression of the host on the worm development were observed. Two hundred metacercariae of G. seoi were orally administered to each mouse, and worms were recovered on days 1, 3, 5, 7, 14 and 21 post-infection (PI). The worm recovery rate was significantly higher in immunosuppressed (ImSP) mice than in immunocompetent (ImCT) mice except on days 1 and 3 PI. The worms attained sexual maturity by day 3 PI with eggs in the uterus, and worm dimensions and the number of uterine eggs continuously increased until day 14 PI in ImSP mice. Worms recovered from ImSP mice were significantly larger in size than those from ImCT mice on days 1 and 3 PI, and the number of uterine eggs was significantly larger in ImSP mice on days 5 and 7 PI. Genital organs such as the ovary, testes, and vitellaria, that were already developed in the metacercarial stage. grew a little in size until day 14 PI. The results show that the C3H/HeN mouse is, though not excellent, a suitable laboratory host for G. seoi.

• Clinical and Experimental Reproductive Medicine
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• 제21권3호
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• pp.315-323
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• 1994

Mammalian oviductal epithelial cells have been known to improve in vitro fertilization and embryonic development. Recently, co-cultured human embryos with the epithelial cells in human genital tract has been reported to improve the pregnancy rate. The purpose of the study was to investigate the effects of the epithelial cells of human genital tract on the development of mouse early embryos and human fertilized oocytes. The epithelial cells of human genital tract were collected from the fallopian tubes which were obtained during hysterectomy in fertile women and from the endometrium during endometrium biopsy. Collected human ampullary cells(HACs) and endometrial cells(HECs) were cultured for 10 days to establish primary monolayer. Second passaged HACs and HECs were obtained by trypsinization were cryopreserved in PBS with 1.5 M DMSO for later use. To investigate the effect when co-cultured with HACs and HECs, we tried to apply strict quality control on mouse embryo, from two cell to blastocyst prior to human trial. The results of quality control were as follows; In Group I (Ham's F10 with 10% FCS), Group IT (co-cultured with HACs) and Group ill (co-cultured with HECs), developmental rates to blastocyst were 63.3%(253/400), 76.0%(304/ 400),74.0%(296/400), respectively. Hatching rates were 36.8%(147/400), 41.80/0(167/400), 38.0%(152/400), respectively(p<0.05). To perform the human IVF, cryopreserved HACs were thawed at 37$^{\circ}C$ waterbath, seeded on the well dish and cultured for 48 hI'S. The pronuclear stage embryos were transferred to the seeded well dish. After 24 hRS, co-cultured embryos were examined and transferred to patient's uterus. The results of human IVF when co-cultured with HACs were that fertilization and developmental rates were 61.8% (256/414), 95.3% (244/256) as compared with 57.2% (279/488) and 94.6%(264/279) in Ham's F10 supplemented with 10% FCS(control). However, 62.9% (161/256) of co-cultured human embryos showed good embryos(no or slight fragmentation) as compared with 53.8 % (150/279) in control(p < 0.05). Pregnancy rate was 40.0% (12/30) when co-cultured with HACs whereas 30.6%(11/36) in control. In conclusions, co-culture system using HACs and HECs improved the developmental and hatching rates of mouse embryo. Also, in human IVF system when co-cultured with HACs, it improved both the quality of human embryos and the pregnancy rate.

• Journal of Pharmaceutical Investigation
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• 제37권5호
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• pp.297-303
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• 2007

The purposes of this study were to clarify the involvement of P-glycoprotein (P-gp) in the efflux of levosulpiride in knockout mice that lack the mdr1a1b gene and to evaluate the relationship between the genetic polymorphisms in MDR1 gene (exon 21) and levosulpiride disposition in healthy Korean subjects. After oral administration ($10\;{\mu}g/g$) of levosulpiride to mdr1a/1b(-/-) and wild-type mice, plasma and brain samples were obtained at 45 min. We also investigated the genotype for MDR1 (exon 21) gene in humans using a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. A single oral dose of 25 mg levosulpiride was administered to 58 healthy subjects, who were based on the MDR1 genotype for the G2677T SNP. Blood samples were taken up to 36 hr after dosing. The concentrations of levosulpiride in mouse plasma and brain were statistically significant difference between the two animal groups (P<0.05). In addition, the average brain-to-plasma concentration ratio (Kp) of levosulpiride was 3.4-fold (P<0.01) higher in the mdr1a/1b(-/-) mice compared with the wild-type mice. We also found that the values of $AUC_{0-{\infty}$, partial AUC ($AUC_{0-4h}$) and $C_{max}$ were significantly different between homozygous 2677TT subjects and the subjects with at least one wild-type allele (GG and GT subjects, P=0.012 for $AUC_{0-{\infty}$; P=0.008 for $AUC_{0-4h}$; P=0.038 for $C_{max}$). The results confirm that levosulpiride is a P-gp substrate in vivo, and clearly demonstrate the effect of SNP 2677G>T in exon 21 of the MDR1 gene on levosulpiride disposition.

• Applied Biological Chemistry
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• 제40권6호
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• pp.490-494
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• 1997

Benzo[a]pyrene (BP)으로 유발한 mouse의 전위암 형성에 대한 astaxanthin 함유 난황 (astaxanthin-containing egg yolk : AEY)의 영향을 연구하였다. Female ICR mouse (6-7 주령, 5 mice/cage, 20 mice/treatment)에게 물과 사료를 자유로이 공급하면서 일주일간 적응시킨 후 다음과 같이 처리를 하였다. BP (2 mg/0.2 ml corn oil)를 각 mouse에 경구투여하기 4 일과 2 일 전에 50 mg AEY, 100 mg AEY, 150 mg AEY, 또는 150 mg control egg yolk (CEY)을 함유하는 phosphate-buffered saline (PBS) 0.2 ml를 경구투여하였다. Control mouse는 0.2 ml PBS와 BP 만 경구투여하였다. 이 과정을 4회 반복하였다. BP를 경구투여 한 일주일 후부터 몸무게와 사료섭취량을 매주 기록하였으며 24주 후에 생존한 모든 mouse에 대해 전위암 생성을 조사하였다. AEY를 처리한 mouse에서는 control mouse나 CEY 처리 mouse에 비해 mouse 당 암의 수가 1/3 정도로 감소되었다. 이와 같은 AEY의 항암효과는 AEY의 처리량에 비례하였다. AEY 처리에 따른 암발생율은 control이나 CEY 처리에 비해 감소되었으나 150 mg AEY 처리에서만 유의성이 있는 감소를 보였다. AEY 처리에 따른 몸무게나 사료 소비량의 감소현상은 볼 수 없었다. 따라서 이 결과는 AEY가 BP로 유발한 mouse의 전위암 발생을 억제함을 암시한다.

• Journal of Pharmaceutical Investigation
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• 제33권1호
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• pp.21-28
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• 2003

Ketorolac tromethamine(KT) is a nonsteroidal agent with potent analgesic and moderate anti-inflammatory activity. The lipid-water partition coefficient of KT was evaluated and KT gel was formulated as a gel containing different pH, different concentrations of polymer (poloxamer 407, carbopol 941), propylene glycol, ethanol and various enhancers. The resulting KT gels were evaluated with respect to their viscosity, in vitro drug permeation rate through hairless mouse skin and stability. In n-octanol and chloroform, the lipid-water partition coefficient of KT was the highest at pH 4 phosphate buffer. The apparent viscosity of KT gel increased with an increase in gel pH, polymer and enhancer concentration. But the apparent viscosity of KT gel decreased with an increase in ethanol concentration. The permeation rate of KT through hairless mouse skin from gels different pH was maximum at pH 4 which is close to KT $pK_{a}$ 3.54. The permeation rate decreased with an increase in polymer, propylene glycol concentration. But the permeation rate increased with an increase in ethanol. The increase of drug concentration from 1 to 3% induced linear increase in permeation rate. The best enhancer was the combination of $Labrasol^{\circledR},\;Transcutol^{\circledR}$, oleic acid and l-menthol. In the accelerated stability test(25, 40 and $50{\circ}C$), pH 5 gel was most stable and pH 4 gel was most unstable for 90 days.

• Radiation Oncology Journal
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• 제20권2호
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• pp.147-154
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• 2002

목적 : 현재까지 방사선에 대한 저산소세포의 감수성을 높이기 위한 많은 실험적 및 임상적 연구가 진행되어 왔으나, 아직 적절한 방법이 개발되지 못한 상태이다. 본 연구에서는 혈관수축 이완 및 혈액점도 저하를 통하여 말초혈류 증가작용을 갖고 있는 Ginkgo biloba extract (GBE)가 종양내 대사상태에 어떠한 변화를 가져오는지를 $^{31}P$ 핵자기 공명 분광법을 통하여 알아보고자 하였다. 방법 : $100\;mm^3$에서 $130\;mm^3$의 섬유육종을 갖는 18마리의 C3H 마우스를 각각 9 마리씩 두 군으로 분리하였다. 한 군은 GBE로 전처치를 하지 않았으며 나머지 다른 한 군은 $^{31}P$ 자기공명분광법을 시행하기 24시간 전에 100 mg/kg의 GBE를 복강내로 투여하여 전처치를 하였다. 우선 각 군에서 $^{31}P$ 자기공명분광법을 실시하여 대조 spectrum을 얻었으며 그 후 100 mg/kg의 GBE를 재투여 하고 약 1시간 후에 $^{31}P$ 자기공명분광법을 다시 실시하였다. 결과 : 전처치를 하지 않은 군에서는 GBE 투여 전의 평균 pH, PCr/Pi, PME/ATP, Pi/ATP, PCr/(Pi+PME) 수치를 GBE 투여 후 1시간에 측정한 값과 비교하였을 때 통계학적인 차이가 없었다. 그러나 전처치를 한 군에서는 GBE 투여 전의 평균 PCr/Pi, Pi/ATP, PCr/(Pi+PME)수치가 0.49, 0.77, 0.17에서 GBE 투여 후에는 0.74, 0.57, 0.28로 변화하였으며 이는 paired-t test상 통계학적으로 의미있는 차이였다. 결론 : 전처치를 한 군에서 GBE의 재투여로 대사상태가 현저히 호전되었으며 이는 간접적으로 GBE 에 의한 방사선감수성의 증가가 혈류 증가 및 이에 따른 대사상태 호전에 기인함을 나타낸다.

• Archives of Pharmacal Research
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• 제21권4호
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• pp.458-464
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• 1998

Search for a new $\alpha$-methylene-$\gamma$-butyrolactone-bearing 6-substituted purine as a potental antitumor agent has led to synthesize seven, hitherto unreported, $5^1$-Methyl-$5^1$-[(6-substituted-9H-purin-9-yl)methyl]-$2^1$-oxo-$3^1$- methylenetetrahydrofurans (H, Cl, l, $CH_3$, $NH_2$, SH, >C=O) (6a-g). These include $5^1$-Methyl-$5^1$-[(9H-purin-9-yl)methyll-$2^1$-oxo-$3^1$ -methylenetetrahydrofurans (6a), $5^1$-Methyl-$5^1$-[(6-chloro-9H-purin-9-yl)methyl]-$2^1$-oxo-$3^1$-methylenetetrahydr ofurans (6b), $5^1$-Methyl-$5^1$-[(6-chloro-9H-purin-9-yl) methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofurans (6c), $5^1$-Methyl-$5^1$-[(6-methyl-9H-purin-9-yl) methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofurans (6d), $5^1$-Methyl-$5^1$-[(9H-adenin-9-yl)methyll-$2^1$-oxo-$3^1$-methylenetetrahydrofurans (6e), $5^1$-Methyl-$5^1$-[(6-mercapto-9H-purin-9-yl) methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofurans (6f) and $5^1$-Methyl-$5^1$-[(9H-hypoxanthin-9-yl)methyll-$2^1$-oxo-$3^1$-methylenetetrahydrof urans (6g) which were made by the Reformatsky-type reaction of ethyl $\alpha$-(bromomethyl) acrylate with the corresponding (6-substituted-9H-purin-9-yl)-2-propanone intermediates (5a-g). These ketone intermediates 5a-g, 1-(9H-purin-9-yl)-2-propanone (5a), 1-(6-chloro-9H-purin-9-yl)-2-propanone (5b), 1-(6-iodo-9H-purin-9-yi)-2-propanone (5c), 1-(6-methyl-9H-purin-9-yl)-2-propanone (5d), 1-(9H-adenin-9-yl)-2-propanone (Se), 1-(6-mercapto-9H-purin-9-yl)-2-propanone (5f), and 1-(9H-hypoxanthin-9-yl)-2-propanone (5g) were directly obtained by the alkylation of the 6-substituted purine bases with the chloroacetone in the presence of $K_2$$CO_3$ (or NaH) under DMF (or DMSO). The preliminary in vitro cytotoxcity assay for the synthetic .alpha.-methylene-y-butyro-lactone compounds (6a-g) were determined against three cell lines (PM-3A, P-388, and K-562) and showed the moderate antitumor activity ($IC_50$ ranged from 1.4 to 4.3 $\mu\textrm{g}$/ml) with the compound $5^1$-methyl-$5^1$ -[(9H-hypoxanthin-9-yl)methyl]-$2^1$-oxo-$3^1$-methylenetetrahydrofuran (6g) showing the least antitumor activity.

• 대한미생물학회지
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• 제22권3호
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• pp.197-208
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• 1987

Chlamydia trachomatis has now shown that this interesting intracellular parasite is a cause of nongonococcal urethritis, infantile pneumonia, pelvic inflammatory disease and epididymitis, in addition to lymphogranuloma venerum and inclusion conjunctivitis. There are several diagnostic methods for C. trachomatis, but the method using monoclonal antibody is the most sensitive and specific. The hybride cell were prepared by fusion of myeloma cell($P_3X_{63}\;Ag_8{\cdot}V_{653}$) of mouse and lymphocyte of mouse(BALB/c) that were immunized with formalin killed C. trachomatis serotype D. The cell mixtures after fusion were dispensed into 640 wells of the 96 well culture plates and continuously cultured in HAT medium for 2 weeks. The supernatants of culture media in 83(13%) wells were reacted with C. trachomatis, which were determined by enzyme-linked immunosorbent assay in 96 well microplate. The clones that secreted antibody to C. trachomatis were cloned by limiting dilution. Only six monoclones secreted antibody to C. trachomatis. The antibody titer of ascitic fluid that collected from same BALB/c mice bearing hybridoma cells was above 1:100,000. These monoclonal antibodies that were IgG reacted with elementary and reticulate bodies of all serotypes(Ba, D, E, F, G, H, J and LGV type-I) using ELISA and indirect immunofluorescence stain, but there were no cross reaction with other bacteria(coagulase negative Staphylococcus, Proteus and E. coli). We concluded these six monoclones secreted the same monoclonal antibody to C. trachomatis. The sensitivity and specificity of the monoclonal antibody compared with Microtrak(confirmatory test of C. trachomatis, Syva) was 100%, respectively.