• Title/Summary/Keyword: In vivo

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Replacement of the in vivo Bioassay for Erythropoietin with the in vitro Bioassay (Erythropoietin in vivo 시험법의 in vitro 대체 시험법 확립)

  • 백상훈;김진만;권기성;박송용;허재욱
    • KSBB Journal
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    • v.18 no.4
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    • pp.255-260
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    • 2003
  • In vivo bioassays for biological medicines have been considered final resort to unequivocally assess the biological activities for them because there are some cases in which the biological activities obtained from in vivo bioassay and in vitro bioassay quite differ each other. The in vivo biological activity of EPO depends on its sialic acid contents which confer microheterogeneity-isoforms to this protein. We have devise a method which consists of a in vitro bioassay using BaF3 cell line and a capillary zone electrophoresis (CZE) for the measurement of the EPO isoform distribution. The biological activity of EPO obtained using in vitro bioassay with BaF3 cell line showed good correlation (C.V.(%) 7.34, 5.85, 8,16, 8.08, 8.8) to EPO content measured either spectrophotometric assay (A280 0.1 % =0.743) or radio immunoassay. The assay validation results of in vitro bioassay with 3 lot of in house EPO showed good results to EPO content measured either in vivo assay or radio immunoassay. and also showed good results the robustness of our method in terms of precision, accuracy, repeatability. The isoform distribution for EPO-BRP (1 : 1 mixture of epoetin-${\alpha}$ and epoetin-${\beta}$, European Pharmacopoeia) by CZE method resulted in isoform 2 through isoform 8. The major peaks in electrophoregram were composed of isoform 3 through 7. Our recombinant EPO (epoetin-${\alpha}$) having equivalent in vivo biological activity showed the isoform distribution of isoform 3 through 9. The major peaks consisted of isoform 4 through 8. The peak area of isoform 4 was always smaller than that of isoform 5. The preparations of recombinant epoetin-${\alpha}$ with lower in vivo biological activity than EPO-BRP showed the isoform 2 through 8 in their electrophoregrams whose major peaks consisted of the isoform 3 through 7. The peak area of isoform 4 was larger than that of isoform 5.

The Study on the Skin Penetration of Cosmetic Ingredient with in vivo Raman Spectroscopy and in vitro Franz Cell (라만 분광 피부 측정기를 이용한 기능성 화장품 성분의 in vivo 피부 투과 측정 및 in vitro 비교 평가 연구)

  • Jeon, Serim;Han, Min-Hee;Chung, Dae-Kyun;Hwang, Jae-Sung
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.40 no.1
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    • pp.1-10
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    • 2014
  • At present, there are few research papers on skin penetration of cosmeceutical ingredients. What is worse is that in vivo studies are hard to find. In this study, we measured skin epidermal penetration of cosmeceutical ingredients using in vivo Raman spectroscopy and compared with the results obtained from experiments using in vitro franz cell. Results showed that ascorbyl-2-glucoside, retinol, retinyl palmitate, and kojic acid were good for penetration ratio in measurement in vitro and retinol, vitamin C, and arbutin were good in measurement in vivo. Among them, retinol was best in skin penetration in vivo experiment using Raman spectroscopy and ascorbyl-2-glucoside was best in skin penetration in vitro experiment using Franz cell system. It is estimated that the differences were originated from the experimental procedures of two different methods; in vivo Raman experiment can be sensitive to the effect of epidermis and dermis as characteristics of matter by estimating the stratum corneum and in vitro measurement is evaluation of material to penetrate skin of hairless mouse. However, most penetration barrier is the stratum corneum, thus it is important to examine movement of material in the stratum corneum. We expect that these results provided useful information for many cosmetic related research.

Differences in in vivo Fluorescence Yield for Netplankton and Nanoplankton Size Classes (Netplankton과 Nanoplankton 크기별 in vivo Fluorescence의 차이)

  • MOON Chang-Ho;LEE Seung-Yong
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.27 no.6
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    • pp.727-732
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    • 1994
  • In the South Sea of Korea, in vivo fluorescence intensity (IVF) and extractable chlorophyll a concentration were measured to determine whether there was significant defference in in vivo fluorescence per unit chlorophyll a (R) between netplankton and nanoplankton size classes (less than $22{\mu}m$). IVF and chlorophyll a were linearly related for both size classes, but R's were significantly different between two size classes. The R of nanoplankton was about 7 times higher than that of netplankton. Therefore, the size dependency of R must be taken into consideration when size fraction of phytoplankton biomass is determined from the measurements of in vivo fluorescence intensity.

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New Record of Three Colpodean Ciliates (Ciliophora: Colpodea) from Korea

  • Kim, Kang-San;Min, Gi-Sik
    • Korean Journal of Environmental Biology
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    • v.33 no.4
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    • pp.375-382
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    • 2015
  • We discovered three soil ciliates of the class Colpodea-Colpoda henneguyi Fabre-Domergue, 1889; C. lucida Greeff, 1888; and Bursaria truncatella $M{\ddot{u}}ller$, 1773 from Obong-ri, Ayajin-ri and Elwang-ri (Korea), respectively. Colpoda henneguyi had the following features: often wider preorally than postorally, size in vivo $60-80{\mu}m{\times}50-70{\mu}m$; extrusomes indistinct in vivo, cylindroid approximately $1{\mu}m$ long; notches caused by deep diagonal groove; yellowish globules on the cortex of the cell; 10-12 postoral kineties; silverline system aspera-type. Colpoda lucida exhibited the following features: broadly reniform, size in vivo $70-90{\mu}m{\times}50-70{\mu}m$; conspicuous extrusomes, $3.5-5{\mu}m$ long in vivo, cylindroid to fusiform; 13-16 postoral kineties; silverline system cucullus-type. Bursaria truncatella had the following features: bursiform, size in vivo $300-470{\mu}m{\times}120-260{\mu}m$; macronucleus coiled with highly variable shapes, $600-1100{\mu}m{\times}30-40{\mu}m$ long in vivo; micronuclei 16-25 in number, approximately $4{\mu}m$ in diameter; extrusomes cylindroid, $3-4{\mu}m$ long in vivo. This is the first report of colpodean ciliates from Korea, and we describe these species based on observations of live and impregnated (protargol and silver nitrate impregnation) specimens.

Correlation between in vitro release and in vivo bioavailability of Propranolol.HCI from Poly(vinyl alcohol) Hydrogel Suppositories (폴리비닐알코올 하이드로겔 좌제로부터 프로프라놀롤의 in vitro 방출과 in vivo 생체이용률간의 상관성)

  • Kim, Ho-Jeong;Ku, Young-Soon
    • Journal of Pharmaceutical Investigation
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    • v.28 no.4
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    • pp.275-282
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    • 1998
  • In order to develop a desirable in vitro release which correlates well with in vivo bioavailability, hollow type suppository containing Propranolol HCl(PPH) powder in the cavity and conventional type suppository with dispersed PPH in the base were prepared. Polyvinyl alcohol (PVA) hydrogel as a base and PPH as a model drug were used for the preparation of suppository. The rates of drug release from the suppositories were studied by Paddle method, Muranish method, Dialysis tubing method and Rotating dialysis cell method. The release profiles from suppositories using the four different release tests were compared. After a rectal administration in rat, the mean $C_{max}$ of hollow type suppository was significantly lower than that of conventional type, but $T_{max}$, $AUC_{0{\to}12}$ and MRT of hollow type were significantly higher 1.6 times, 1.2 times and 1.9 times than those of conventional type, respectively. The computer program was used to simulate plasma concentration from in vitro released amounts of drug and in vivo pharmacokinetic parameters. Based on comparison of the simulated bioavailability from computer program with experimental bioavailability in rat we have found out in vitro release test which correlates well with in vivo bioavailability. Our results have shown the best correlation between in vitro release and in vivo bioavailability in PPH-PVA hydrogel hollow type suppository for the paddle method and conventional type suppository for the rotating dialysis cell method. In this work we propose that PPH-PVA hydrogel suppository shows in vitro-in vivo correlation. This data should help to optimize the formulation of the drug and provide a basis for quality control procedures.

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Effects of Iron, Chelators and Nitrate Concentration on in vivo Fluorescence and Nitrate Reductase of the Red Tide Organism Amphidinium carterae

  • Yang, Sung-Ryull;Song, Hwan-Seok;Pae, Se-Jin;Huh, Sung-Hoi
    • Journal of the korean society of oceanography
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    • v.34 no.1
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    • pp.49-57
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    • 1999
  • A red tide organism, Amphidinium carterae was incubated under different iron/chelator and nitrate concentrations to investigate the factors controlling the growth. The chelation capacity played a critical role in regulating the nitrate reductase (NR) activity and in vivo fluorescence of this organism. However, there was a significant difference between the NR activity and in vivo fluorescence in response to trace metals and chelator treatments. In vivo fluorescence was the highest in FeEDTA 10 ${\mu}$M treatments and the lowest in DTPA 10 ${\mu}$M treatments. This indicates that the availability of the trace metal is important in regulating the in vivo fluorescence of this photosynthetic microalgae In contrast, NR activity showed the highest values in trace metal enriched treatments, and trace metal + DTPA treatments showed fairly high NR activities. This suggests that DTPA treatment did not hinder the NR activity as much as it did in vivo fluorescence. In vivo fluorescence and NR activity increased with nitrate concentration of up to 50 ${\mu}$M and remained relatively constant or the rate of increase decreased above that concentration, indicating that initial nitrate concentration of higher than a certain level would not accelerate the growth of A. carterae. Further investigation is needed to elucidate the reason for the difference in timing sequence between the NR and in vivo fluorescence in response to different metal treatments and chelation capacity.

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A Study on the Source of Reductants for Nitrate Reduction in Rice (Oryza sativa cv. Tongil) Roots (벼(Oryza sativa cv. Tongil) 뿌리에 있어서 Nitrate 환원에 필요한 환원력의 공급원에 관한 연구)

  • Chang, Nam-Kee;Choe, Hong-Gwan
    • The Korean Journal of Ecology
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    • v.7 no.2
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    • pp.85-90
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    • 1983
  • There was a decrease in nitrate reductase activity (NRA) measured in vivo in rice roots (Oryza sativa cv. Tongil) grown in anaerobic culture solution. But it was reversed by addition of malonate to the in vivo nitrate reduction assay medium. Malonate increased the in vivo NRA during 2-5 hours incubation and decreased it in longer incubation hours. In vivo NRA was stimulated by addition of NaHCO3 to the assay medium, but not by Na2CO3. The stimulation of NRA by NaHCO3 was not observed in shoot removed rice roots. It is suggested that CO2 from NaHCO3 is carboxylated by phosphoenol pyruvate carboxylase, results in increasing the malate contents in the roots, and stimulates the in vivo NRA. NADH needed in nitrate reduction is supported by malate oxidation. In rice roots, it seems probable that malate oxidation in the mitochondria is more important to nitrate reduction than malae oxidation in cytoplasm.

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ULTRASTRUCTURAL COMPARISON OF BOVINE BLASTOCYSTS DEVELOPED IN VIVO AND IN VITRO

  • Ohboshi, S.;Nakamichi, R.;Hanada, K.;Zhao, J.;Hattori, M.;Fujihara, N.;Umetsu, R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.8 no.6
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    • pp.599-605
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    • 1995
  • The ultrastructures of in vitro-derived bovine blastocysts have been compared with those of blastocysts obtained from a superovulated cow. In vivo blastocysts obtained from the uterus showed well-differentiated features, while in vitro-derived embryos, which were developed from in vitro fertilized ovum, showed insufficient cellular organizations. In vitro-derived embryos contained many undefined cellular organizations in the perivitelline spaces compared with in vivo-derived blastocysts. Other features of in vivo and in vitro blastocysts were characterized by differential development of microvilli projection into blastocoele from the surface of the trophoblast cells. The conceivable reason for the difference between in vivo and in vitro developments of bovine embryos is that it is likely that in vitro culture system adopted in the present experiment may not be sufficient for better embryonic development.

The Emergence of Behavioral Testing of Fishes to Measure Toxicological Effects

  • Brooks, Janie S.
    • Toxicological Research
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    • v.25 no.1
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    • pp.9-15
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    • 2009
  • Historically, research in toxicology has utilized non-human mammalian species, particularly rats and mice, to study in vivo the effects of toxic exposure on physiology and behavior. However, ethical considerations and the overwhelming increase in the number of chemicals to be screened has led to a shift away from in vivo work. The decline in in vivo experimentation has been accompanied by an increase in alternative methods for detecting and predicting detrimental effects: in vitro experimentation and in silico modeling. Yet, these new methodologies can not replace the need for in vivo work on animal physiology and behavior. The development of new, non-mammalian model systems shows great promise in restoring our ability to use behavioral endpoints in toxicological testing. Of these systems, the zebrafish, Danio rerio, is the model organism for which we are accumulating enough knowledge in vivo, in vitro, and in silico to enable us to develop a comprehensive, high-throughput toxicology screening system.