• 대한수의학회지
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• 제38권3호
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• pp.589-594
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• 1998

The in vitro screening tests against the in vitro cultivated $L_3$ of Ascaris suum (in vitro $L_3$), which were cultivated from the embryonated egg to third-stage larva on 7 days in culture(DIC) and the in vivo rat's lung-derived $L_3$ of Ascaris suum (in vivo $L_3$), which were recovered from the lungs of rat on 7 days after infection, carried out in order to compare the anthelmintic efficacy of in vitro $L_3$ and that of in vivo $L_3$ in RPMI medium 1640 with 5% bovine calf serum. And also a screening test of efficacy against adult worms of Trichuris suis performed. The efficacies of screening tests were as follows : 1. The screening efficacies of abamectin and ivermectin against the in vitro $L_3$ were all 100% at the 10ppm concentration in RPMI medium 1640 on 5 DIC. 2. The screening efficacies of abamectin and ivermectin against the in vivo $L_3$ were all 100% at the 20ppm on 5 DIC or at 40ppm on 3 DIC. 3. The screening efficacies of abamectin and ivermectin against the adult worms of Trichuris suis were all 100% at 20ppm on 4 DIC. And therefore, the in vitro cultivated $L_3$ of Ascaris suum were used in the screening test as well as the in vivo rat's lung-derived $L_3$ of Ascaris suum. And also the adult worms such as Trichuris suis and filaroids which is small size and difficult to cultivate to vitro, were used in the screening test in vitro.

• 대한화학회지
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• 제52권5호
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• pp.495-502
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• 2008

• 농약과학회지
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• 제3권3호
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• pp.37-44
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• 1999

In vivo와 in vitro 생물검정법에서의 활성관계를 조사하고 또한 보다 효율적인 in vitro 검정법을 개발하기 위하여 작용기구가 다른 두 계열 살균제 즉 sulphamide계 (dichlofluanid와 tolylfluanid)와 dicarboximide계 살균제 (iprodione, vinclozolin 및 procymidone)을 시험약제로 하여 본 실험을 수행하였다. Dichlofluanid, tolylfluanid, iprodione, vinclozolin procymidone의 토마토 유묘에서의 병 방제효과는 유사하였다. 그러나 이들 살균제에 대해 in vitro 생물검정법으로 항균활성을 비교한 결과, sulphamide계 살균제는 포자발아 억제실험에서 높은 활성을 나타낸 반면에, dicarboximide계 살균제는 균사생장 억제실험에서 항균활성이 더 우수하였다. 그러므로 in vivo 병 방제효과가 유사하더라도 작용기구가 다르면 in vitro 생물검정법에서의 항균활성은 다르게 나타남을 알 수 있었다. 또한 포자현탁액을 넣은 96-well microtiter plate에 약제를 처리하고 3시간 배양 후 배양액을 제거하고 다시 새 배지를 넣어 배양한 후 생장억제를 조사한 실험 (microtiter plate method I, MPM I)에서는 살균제들의 포자발아 억제실험과 유사한 경향의 활성을 보였다. 96-well microtiter plate에서 포자현탁액을 6시간 동안 배양하여 포자를 발아시킨 후에 약제를 처리한 실험(microtiter plate method II, MPM II)에서의 항균활성은 균사생장 억제효과와 일치하는 경향을 보였다. 따라서 기존의 in vitro 항균활성 실험보다 편리하고 효율적인 MPM I과 II 방법은 각각 포자발아와 균사생장 억제실험을 대신할 수 있으리라 생각되었다.

• Toxicological Research
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• 제16권2호
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• pp.109-116
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• 2000

The purposes of our study were to optimize the conditions of the screening and testing methods for endocrine disruptors, to characterize these assays using several compounds with well-defined endocrine activity, and to compare the sensitivity between these assays currently undergoing validation. Two in vitro test systems, MCF-7 cells proliferation (E-screen assay) and competitive binding to estrogen receptors (ER) were selected to evaluate the estrogenic effects. 17$\beta$-Estradiol (E2) and diethylstilbestrol (DES) were used as a positive control in vitro test. Also, E2 and ethinyl estradiol (EE) were used as a positive control in vivo uterotrophic assay. In in vitro test, E2 and DES showed a strong estrogenic response at concentration of 1.0 nM. In uterotrophic assay, E2 (0.3 $\mu\textrm{g}$/kg) and EE (0.3 $\mu\textrm{g}$/kg) produced a significant increase in uterus and vagina weight in both immature and ovariectomized rats. Although we did not com-pared the specificity between in vivo and in vitro assays, these assay systems may serve as a good tool for endocrine disruptors screening methods. Our data indicate that these assay systems exhibit some difference in their sensitivity to the same estrogenic compounds. Therefore, as a first rapid screening assay for estrogenic activity qf unknown chemicals, at least two assay systems should probably be carried out with a view of high sensitivity and standardization conditions. Also, a careful validation tests are necessary to obtain a reasonable degree of reproducibility.

• Toxicological Research
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• 제35권4호
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• pp.343-351
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• 2019

Many in vitro developmental toxicity assays have been proposed over several decades. Since the late 1980s, we have made intermittent attempts to introduce in vitro assays as screening tests for developmental toxicity of inhouse candidate products. Two cell-based assays which were developed two decades apart were intensively studied. One was an assay of inhibitory effects on mouse ascites tumor cell attachment to a concanavalin A-coated plastic sheet surface (MOT assay), which we studied in the early days of assay development. The other was an assay of inhibitory effects on the differentiation of mouse embryonic stem cell to beating heart cells (EST assay), which we assessed more recently. We evaluated the suitability of the assays for screening in-house candidates. The concordance rates with in vivo developmental toxicity were at the 60% level. The EST assay classified chemicals that inhibited cell proliferation as embryo-toxic. Both assays had a significant false positive rate. The assays were generally considered unsuitable for screening the developmental toxicity of our candidate compounds. Recent test systems adopt advanced technologies. Despite such evolution of materials and methods, the concordance rates of the EST and MOT systems were similar. This may suggest that the fundamental predictivity of in vitro developmental toxicity assays has remained basically unchanged for decades. To improve their predictivity, in vitro developmental toxicity assays should be strictly based on elucidated pathogenetic mechanisms of developmental toxicity.

• 대한화장품학회지
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• 제29권2호
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• pp.37-65
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• 2003

새로운 주름개선 효능물질을 개발하기 위하여 여러 가지 세포배양 모델을 이용한 in vitro 효능물질의 검색방법을 이용하여 후보물질을 도출하고 있다. 이처럼 In vitro에서 선별된 주름개선 후보물질은 사람피부에 직접 도포 하였을 때 in vitro에서 와 는 달리 주름개선 효능이 없는 경우가 많이 있다. 즉 in vitro 검색결과와 사람피부에서의 효능은 현실적으로 큰 차이를 보인다는 것이다. 그러나 모든 효능 후보물질을 사람 피부에 직접 검사할 수는 없는 현실적 어려움이 있다. 이러한 애로점을 해결하기 위하여, 동물모델이나, 인공피부배양모델 등을 이용하는 추세이다. 이번 강의에서, 본인은 동물모델과 in vitro에서 효능 측정방법을 상세하게 리뷰 하고 각 방법에 대해 토의할 것이며, 임상 전 시험 결과를 보고 할 것이다. 그 중 특히 동물 모델은 주름개선 효능물질의 효능을 평가하는데 좋은 대상이 될 것이다.

• 대한화장품학회지
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• 제34권3호
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• pp.183-188
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• 2008

미백제를 선발하기 위해 주로 사용하는 현재의 방법은 in vitro 타이로시네이즈 활성 및 항산화능을 측정하는 것이다. 이 결과에 기초하여 다음 단계인 멜라노사이트에서의 멜라닌 생성량을 측정한다. 세포 내의 멜라닌 생성량 측정 법은 시간, 인력 및 숙련도가 요구된다. 따라서 초기 선발 방법의 신뢰성이 중요하다. 200개 중국시료 중 측정범위 내에서 세포독성이 없는 34개를 대상으로 세포 내 멜라닌량, 타이로시네이즈 활성, 항산화능의 상관관계를 조사하였다. 조사결과 직선의 상판관계를 확인할 수 없었다. 이 결과는 현재 선발방법의 한계 및 새로운 방법이 필요함을 보여주었다.

• Journal of Pharmaceutical Investigation
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• 제38권3호
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• pp.183-189
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• 2008

The present study compared the feasibility of Caco-2 and MDCK cells as an efficient in-vitro model for the drug classification based on Biopharmaceutics Classification System (BCS) as well as an in-vitro model for drug interactions mediated by P-gp inhibition or P-gp induction. Thirteen model drugs were selected to cover BCS Class I{\sim}IV$ and their membrane permeability values were evaluated in both Caco-2 and MDCK cells. P-gp inhibition studies were conducted by using vinblastine and verapamil in MDCK cells. P-gp induction studies were also performed in MDCK cells using rifampin and the P-gp expression level was determined by western blot analysis. Compared to Caco-2 cells, MDCK cells required shorter period of time to culture cells before running the transport study. Both Caco-2 and MDCK cells exhibited the same rank order relationship between in-vitro permeability values and human permeability values of all tested model compounds, implying that those in-vitro models may be useful in the prediction of human permeability (rank order) of new chemical entities at the early drug discovery stage. However, in the case of BCS drug classification, Caco-2 cells appeared to be more suitable than MDCK cells. P-gp induction by rifampin was negligible in MDCK-cells while MDCK cells appeared to be feasible for P-gp inhibition studies. Taken all together, the present study suggests that Caco-2 cells might be more applicable to the BCS drug classification than MDCK-cells, although MDCK cells may provide some advantage in terms of capacity and speed in early ADME screening process.

• Archives of Pharmacal Research
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• 제22권2호
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• pp.137-142
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• 1999

Anti-ulcer activity of newly synthesized acylquinoline derivatives was investigated. For the in vitro screening, the effects of compounds on gastric $H^{+}/K^{+}$ ATPase isolated from hog and rabbit were examined. Among them, AU-090, AU-091, AU-254, AU-413 and AU-466 exhibited good in vitro activity on both enzymes. To correlate the in vitro activity with in vivo action, the effects of the compounds on the basal gastric acid secretion were studied. Some derivatives showed considerable anti-secretory activities, and AU-413 was selected for further studies. AU-413 protected gastric damage induced by either ethanol or NaOH dose dependently when given orally. $ED_{50}$ values of 12 mg/kg, p.o. (ethanol) and 41 mg/kg, p.o. (NaOH) were obtained. In addition, histamine-stimulated gastric secretion was reduced upon AU-413 administration. Taken together, newly synthesized acylquinoline derivatives, especially AU-413, is worthy of further investigation to be developed as an anti-ulcer agent.

• Clinical and Experimental Reproductive Medicine
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• 제51권1호
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• pp.85-90
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• 2024

Objective: The purpose of this study was to compare fresh and frozen-thawed euploid blastocyst transfer protocols following preimplantation genetic screening (PGS) in cases of advanced maternal age. Methods: A total of 330 patients were examined retrospectively. PGS was performed on the embryos of 146 patients for whom fresh transfers were chosen. In contrast, frozen-thawed euploid single embryo transfer (ET) was selected after PGS for 184 patients, and their embryos were vitrified. The percentage of euploid embryos and rates of implantation, pregnancy, and pregnancy continuity, as well as clinical and biochemical abortion rates, were compared. Results: The numbers of retrieved oocytes, metaphase II oocytes, and fertilized ova were greater in the frozen-thawed group. The percentages of euploid embryos were comparable between the fresh and frozen-thawed groups (32% vs. 34.8%, respectively). The rates of implantation (46.6%vs. 62.5%), pregnancy (50% vs. 66.8%), ongoing pregnancy (38.4% vs. 53.8%), and live birth percentage (37.0% vs. 53.8%) were significantly higher in the frozen-thawed group. However, no significant differences were found in the clinical and biochemical abortion rates. Conclusion: The use of frozen-thawed single euploid ET is associated with increased implantation and pregnancy rates compared to fresh single euploid ET with PGS.