• Title/Summary/Keyword: in vivo and in vitro test

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Immunological significance of recombinant VP2 and VP3 proteins of aquabirnavirus in olive flounder, Paralichthys olivaceus

  • Kim, Hyoung Jun;Kim, Ki Hong;Park, Jeong Su;Lee, Haeng Lim;Kwon, Hyuk Chu;Kwon, Se Ryun
    • Journal of fish pathology
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    • v.28 no.2
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    • pp.93-98
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    • 2015
  • Full-length recombinant VP2 and VP3 proteins of aquabirnavirus isolated from olive flounder were expressed successfully in E. coli expression system. After rats were immunized with these proteins, antisera were used for in vitro and in vivo neutralization test. In in vitro test, VP2 antibody titers were higher than that of VP3. In in vivo assays, fish challenged with aquabirnavirus neutralized with VP2 antibody survived longer than other fish.

In vitro test using chorioallantoic membrance vascular assay to assess the irritancy potential of surfactants (CAMVA(Choriollantoic Membrane Vascular Assay)를 이용한 계면활성제의 자극 평가에 관한 연구)

  • Go, Jae-Suk;An, Su-Seon;Park, Jong-Ho
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.21 no.1
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    • pp.67-83
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    • 1995
  • Skin irritations accompany the series of complex, physical and chemical changes. Alternative methods which reflect the biological phenomenon more sensitively are necessary for the purpose of reducing the animal tests and improving the accuracy and reliability. Chorioallantoic membrane (CAM) vascular assay is a in vitro method which utilizes the chorioallantoic membrane of hen's fertilized egg of about 10 days old. Test substances are placed directly onto the surface of the CAM and 1 hour later the CAM vasculature is subjectively evaluated to determine with a chemically related-injurious reponse including the appearance of haemorrage, congestion, coagulation, and so on. In this research, using the various surfactants, the correlations of CAMVA with in vivo models (intradermal safety test and human primary irritation test) were investigated. And CAMVA closely correlated to intradermal safety test (r=0.89) and human primary irritation test (r=0.90). From the result, it seems that CAMVA can also be used as a method for predicting the skin irritaions.

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Toxicological Test Methods and AChE Inhibition of Organophosphorus Acaricides of Twospotted Spider Mite, Tetranychus urticae (점박이응애의 독성 시험방법과 유기인계 살비제 AChE 활성저해에 관한 연구)

  • 김도익;이승찬
    • Korean journal of applied entomology
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    • v.31 no.1
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    • pp.1-6
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    • 1992
  • These studies were carried out to compare toxicological test methods of two spotted spider mite, Tetranychus urticae Koch,; and to investigate relationship between in vivo resistant level of highly acaricide-selected population, and in vitro insensitivity of the AChE in the same population to carbophenothion and ethion. The slide dip method (CV = 8.7%) was of more accuracy and suitability than that of the leaf dip method(CV=12.2%) and leaf disc method (CV= 13.6 %) in determination of the resistant levels of twospotted spider mite to acaricides. The slide dip method also had the advantages of simple treatment with different populations on a slide at the same time, standardization of post-treatement conditions and living plants exclud¬ed from the test. Even though the topical application method(CV =8.1 %) showed high accuracy, it had the demerits of the much time consuming, need of expensive equipment and difficulty of test manipulation. For a limited time, the 22nd successive carbophenothion-selected population of two-spotted spider mite showed 156- and 128-fold resistant levels to carbophenothion and ethion(both alPs), respectively. However, the 24th successive ethion-selected population revealed 64.1- and 65-fold resistant levels to ethion and carbophenothion, respectively. In the inhibition of AChE activity, the carbophenothion-selected population showed 3.3- and 2.7-fold insensitivity in AChE activity to carbophenothion and ethion, respectively. Likewise, the ethion -selected population exhibited 3- and 2.6-fold insensitivity in AChE activity to carbophenothion and ethion, respectively, as compared with that of susceptible population. As a result, a good relation was recognized between in vivo resistance to organophosphorous acaricides and in vitro insensitivity of the AChE to corresponding inhibitors.

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Genotoxicity Study of HM10411, Recombinant Human Granulocyte Colony Stimulating Factor (재조합 인과립구 콜로니 자극인자 HM10411의 유전독성 연구)

  • 권정;이미가엘;홍미영;조지희;정문구;권세창;이관순
    • Biomolecules & Therapeutics
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    • v.10 no.4
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    • pp.268-273
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    • 2002
  • Mutagenic potential of HM10411 (recombinant human granulocyte colony stimulating factor) was evaluated by bacterial reverse mutation test, in vitro chromosome aberration test and in vivo micronucleus test. The bacterial reverse mutation test was performed using the histidine auxotroph strains of Salmonella typhimurium TA100, TA1535, TA98, TA1537 and tryptophan auxotroph strain of Escherichia coli WP2 uvrA. The negative results of the bacterial reverse mutation test suggest that HM10411 does not induce mutation, in the genome of Salmonella typhimurium and E. coli under the conditions used. In addition, it has little clastogenicity either in vitro chromosome aberration test or in vivo micronucleus test. For in vitro chromosomal aberration test, Chinese hamster lung(CHL) cells were exposed to HM10411 of 23, 46 or 92 $\mu\textrm{g}$/ml for 6 or 24 hours in the absence and for 6 hours in the presence of metabolic activation system. There was no significant increase in the number of aberrant metaphase in HM 10411-treated groups at any dose levels both in the presence and absence of metabolic activation system. The micronucleus test was carried out using specific pathogen free(SPF) 7-week old male ICR mice, The test item, HM10411 was intraperitoneally administered at 1150, 2300 or 4600 $\mu\textrm{g}$/kg once a day for 2 consecutive days. There was no significant increase in the frequencies of micronucleated polychromatic erythrocytes(PCEs) at any treated groups compared with negative control group. Therefore, these results demonstrate that the test item, HM10411, was not mutagenic under the condition of these studies.

Affirmative Effect of Hwaweo-jeon (Huayu-jian) in Osteoblast Cells and Tibia Fracture-induced Mice (화어전(化瘀煎)이 조골세포 및 경골골절 유발 생쥐의 골유합에 미치는 영향)

  • Lee, Soo-Hwan;Parichuk, Kira;Cha, Yun-yeop
    • Journal of Korean Medicine Rehabilitation
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    • v.30 no.1
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    • pp.13-29
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    • 2020
  • Objectives This study was performed to decide the bone union effect of Hwaweo-jeon on tibia fractured mice. Methods In this study, laboratory experiments were implemented by the stage of in vitro and in vivo. In in vitro, MC3T3-E1 cells were treated with various concentration of Hwaweo-jeon extract (HWJ). To investigate effect of HWJ for osteoblast, relative mRNA expression of 5 substances (alkaline phosphatase [ALP], runt-related transcription factor 2 [Runx2], osteocalcin [OCN], osterix [OSX] and collagen type II alpha 1 chain [Col2a1]) was used as a marker of osteogenesis. In order to determine HWJ's effect for fracture healing, relative gene expression level of ALP, Runx2, OCN, OSX and Col2a1 were used to find out the influence to osteoblast. Furthermore, receptor activator of nuclear factor kappa-B ligand and osteoprotegerin relative mRNA expression were used to estimate the impact to osteoclast. Also, X-ray was used for the purpose of identifying bone union in tibia-fracture mouse model. Results In in vitro experiment, most part of relative mRNA expression were increased compared to control group. In in vivo and in vitro experiment, HWJ induced osteoblast activitation by verifying relative mRNA expression of 5 substances. And in vivo experiment, we can also identify that HWJ triggered osteoclast activation during early stage of tibia fracture. Furthermore, X-ray pictures show noticeable recovery of tibia fracture. Conclusions HWJ extract promotes bone union by facilitating the osteoblast. But, HWJ may occur liver & kidney toxicity over specific concentration. Therefore, when HWJ is applied to human body, doctors have to follow up the liver function test & renal function test of patient.

In vitro Biocompatibility Evaluation of Biomaterial-elution Using Inflammatory Cell Lines (염증세포주를 이용한 생체재료 용출물의 체외 생체적합성 평가)

  • Shin, Youn-Ho;Song, Kye-Yong;Seo, Min-Ji;Kim, Sung-Min;Park, Jung-Keug;Kim, Dong-Sup;Park, Ki-Jung;Hur, Chan-Hoi;Cha, Ji-Hun;Seo, Young-Kwon
    • KSBB Journal
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    • v.26 no.3
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    • pp.248-254
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    • 2011
  • Various biometerials have been researched and have been developed for treatment of some disease through transplantation to body. They have been evaluated by in vitro cytotoxicity test using some skin-derived cell lines for prediction of their biocompatibility in vivo. However, the results of experiments using mesenchymal or epithelial cells could not be considered in vivo immune reaction. In this study, we evaluated the biomaterial-elution (elute from high density polyethylene film) using some cell lines (L929, Jurkat, U937) in vitro, and then that results were compared with in vivo results from guinea pig sensitization test. In sensitization test, saline and elution of syringe could not induce erythema, but only DNCB (hypersensitive chemical) induce erythema at guinea pig sensitization test. In cell experiment, the cytotoxicity results of inflammatory cells (Jurkat; T lymphocyte, U937; monocyte) was no difference with L929 (fibroblast) in the overall trend. However, inflammatory cell lines were only secreted inflammatory cytokine (TNF-${\alpha}$, INF-${\gamma}$) in some materials (biomateriallution, FAC, DNCB). And the biomaterial-elution did not have toxicity to the cells, but it induced the inflammatory cytokines in inflammatory cell lines only. So, we were predicted inflammatory reaction through the cytokine resultes of inflammatory cell lines, and it was more correlated with in vivo results than cytotoxicity test. Therefore, we suggested that the inflammatory cytokine assay using inflammatory cell lines are more effective method in vitro for evaluation of biocompatibility of biomaterials or chemicals.

Allergy tests in clinical practice (알레르기 검사의 실제)

  • Lee, Jaechun
    • Journal of Medicine and Life Science
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    • v.15 no.1
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    • pp.1-5
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    • 2018
  • Allergy is conditions when a hypersensitivity reaction happens with a certain element, called as an allergen, which is commonly not reactive to ordinary individuals. Allergic diseases involve various organs or systems in the body. The purpose of allergy tests is to make a diagnosis of allergic diseases and to identify the affecting allergens. In vivo tests, more relevant in clinical situation, include skin test, patch test and provocation test. In in vitro tests, there are specific IgE test, histamine releasing assay, and lymphocyte activation test, safer and more objective than in vivo tests. In the view point of clinical practice, skin test, provocation test, total IgE test and specific IgE test were reviewed in depth.

Genotoxicological Safety of the Gamma-Irradiated Korean Red Ginseng In vitro (감마선 조사 홍삼의 안전성에 관한 유전독성학적 연구)

  • 조성기;육홍선;변명우
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.25 no.3
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    • pp.491-496
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    • 1996
  • This experiment was performed to determine the safety of the Korean red ginseng irradiated with gamma rays with respect to genotoxicity. Ethanol extracts of the 5 and 10 kGy gamma-irradiated red ginseng were examined in two short-term in vitro tests : (1) Salmonella typhimurium reversion assay(Ames test) in strain TA 98, TA 100 and TA 102 (2) Micronucleus test in cultured Chinese hamster ovary(CHO) cells. No mutagenicity was detected in the two assays with or without metabolic activation. It was suggested that the Korean red ginseng irradiated with gamma rays did not cause genotoxicity in vitro. Further tests of genotoxicity in vivo, chronic and reproductive toxicity should be carried out to determine whether it is safe to irradiate Korean red ginseng with practical doses of gamma rays.

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Skin corrosion and irritation test of sunscreen nanoparticles using reconstructed 3D human skin model

  • Choi, Jonghye;Kim, Hyejin;Choi, Jinhee;Oh, Seung Min;Park, Jeonggue;Park, Kwangsik
    • Environmental Analysis Health and Toxicology
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    • v.29
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    • pp.4.1-4.10
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    • 2014
  • Objectives Effects of nanoparticles including zinc oxide nanoparticles, titanium oxide nanoparticles, and their mixtures on skin corrosion and irritation were investigated by using in vitro 3D human skin models ($KeraSkin^{TM}$) and the results were compared to those of an in vivo animal test. Methods Skin models were incubated with nanoparticles for a definite time period and cell viability was measured by the 3-(4, 5-dimethylthiazol-2-yl)-2.5-diphenyltetrazolium bromide method. Skin corrosion and irritation were identified by the decreased viability based on the pre-determined threshold. Results Cell viability after exposure to nanomaterial was not decreased to the pre-determined threshold level, which was 15% after 60 minutes exposure in corrosion test and 50% after 45 minutes exposure in the irritation test. IL-$1{\alpha}$ release and histopathological findings support the results of cell viability test. In vivo test using rabbits also showed non-corrosive and non-irritant results. Conclusions The findings provide the evidence that zinc oxide nanoparticles, titanium oxide nanoparticles and their mixture are 'non corrosive' and 'non-irritant' to the human skin by a globally harmonized classification system. In vivo test using animals can be replaced by an alternative in vitro test.

Studies on the anti-coagulant component of Loranthus yadoriki

  • Lee, Sun-Kyung;Song, Hee-Sun;Yoo, Eun-A;Yang, Hyun-Ok
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2003.11a
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    • pp.71-71
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    • 2003
  • Methanol extract of Loranthus yadoriki showed the prolongation effect of bleeding time in vivo using mice in dose dependent manner. From the MeOH extract of Loranthus yadoriki, compound-A was isolated by the activity guided isolation method using silicagel column chromatography. The anti-coagulant activity was evaluated by the bleeding time test in vivo and plasma recalcification time test in vitro. Compound A showed moderate anti -coagulant activity on plasma recalcification time in vitro.

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