• Title/Summary/Keyword: In vitro testing

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Blood-brain barrier-on-a-chip for brain disease modeling and drug testing

  • Cui, Baofang;Cho, Seung-Woo
    • BMB Reports
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    • v.55 no.5
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    • pp.213-219
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    • 2022
  • The blood-brain barrier (BBB) is an interface between cerebral blood and the brain parenchyma. As a gate keeper, BBB regulates passage of nutrients and exogeneous compounds. Owing to this highly selective barrier, many drugs targeting brain diseases are not likely to pass through the BBB. Thus, a large amount of time and cost have been paid for the development of BBB targeted therapeutics. However, many drugs validated in in vitro models and animal models have failed in clinical trials primarily due to the lack of an appropriate BBB model. Human BBB has a unique cellular architecture. Different physiologies between human and animal BBB hinder the prediction of drug responses. Therefore, a more physiologically relevant alternative BBB model needs to be developed. In this review, we summarize major features of human BBB and current BBB models and describe organ-on-chip models for BBB modeling and their applications in neurological complications.

RNase Resistant RNA in the Egg of Xenopus laevis: I. RNA Extraction and in Vitro Labeling

  • Chung, Hae-Moon
    • The Korean Journal of Zoology
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    • v.20 no.1
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    • pp.9-18
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    • 1977
  • RNA was extracted from the eggs of Xenopus laevis to do preliminary experiments before testing the possibility that if RNase resistant RNA molecules exist in the amphibian egg. Chromatography on Sephadex G-100 column indicated 3 peaks consistently. Only high molecular weight RNA species eluted in the first peak were labeled in vitro using $^{3}H$-dimethyl sulfate to eliminate the possible contribution of base paired oligonucleotides from tRNA. By this method, high specific activity could be obtained and the attached methyl groups were quite stable.

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Screening and Characterization of Lactic Acid Bacteria Strains with Anti-inflammatory Activities through in vitro and Caenorhabditis elegans Model Testing

  • Lee, Hye Kyoung;Choi, Sun-Hae;Lee, Cho Rong;Lee, Sun Hee;Park, Mi Ri;Kim, Younghoon;Lee, Myung-Ki;Kim, Geun-Bae
    • Food Science of Animal Resources
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    • v.35 no.1
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    • pp.91-100
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    • 2015
  • The present study was conducted to screen candidate probiotic strains for anti-inflammatory activity. Initially, a nitric oxide (NO) assay was used to test selected candidate probiotic strains for anti-inflammatory activity in cultures of the murine macrophage cell line, RAW 264.7. Then, the in vitro probiotic properties of the strains, including bile tolerance, acid resistance, and growth in skim milk media, were investigated. We also performed an in vitro hydrophobicity test and an intestinal adhesion assay using Caenorhabditis elegans as a surrogate in vivo model. From our screening, we obtained 4 probiotic candidate lactic acid bacteria (LAB) strains based on their anti-inflammatory activity in lipopolysaccharide (LPS)-stimulated RAW 264.7 cell cultures and the results of the in vitro and in vivo probiotic property assessments. Molecular characterization using 16S rDNA sequencing analysis identified the 4 LAB strains as Lactobacillus plantarum. The selected L. plantarum strains (CAU1054, CAU1055, CAU1064, and CAU1106) were found to possess desirable in vitro and in vivo probiotic properties, and these strains are good candidates for further investigations in animal models and human clinical studies to elucidate the mechanisms underlying their anti-inflammatory activities.

Development of Automated Diffusion Cell for Determining In Vitro Drug Release from Transdermal Device (경피흡수제형의 in vitro 약물방출실험을 위한 연속확산 장치의 개발)

  • Byun, Young-Rho;Choi, Young-Kweon;Jeong, Seo-Young;Kim, Young-Ha
    • YAKHAK HOEJI
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    • v.34 no.3
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    • pp.161-165
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    • 1990
  • An automated, simple, and reliable method was developed for determining in vitro drug release rate from transdermal delivery dosage forms. The patch is held in position in the heating block by sandwiching it between the middle plate and the bottom plate of diffusion cell. The dissolution profile of the commercially available transdermal scopolamine patch was determined over a 72-h period, and the results were compared with those obtained with other methods; paddle-over-disk method, reciprocating method, and diffusion cell method. It was demonstrated that the flow-through method is equivalent in terms of release rate profile and accumulated released drug amount over the lifetime of the dosage form tested. Also this method is simple, reliable and reproducible. Therefore, this technique can be used in a quality control for assuring product uniformity.

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In vivo and in vitro efficacy of florfenicol, terbinafine, and mometasone furoate topical otic solution for the treatment of canine otitis externa

  • Bae, Seulgi;Jin, Yunseok;Oh, Taeho
    • Korean Journal of Veterinary Research
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    • v.61 no.4
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    • pp.37.1-37.6
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    • 2021
  • This study evaluated the efficacy of a new, single-dose otic solution combining florfenicol, terbinafine, and mometasone furoate for the treatment of canine otitis externa (OE) in vitro and in vivo. Forty-one client-owned dogs with OE were included in the study and divided into a treatment group that received the test solution and a negative control group that received a normal saline solution. On day 0, the dogs were treated either with the test or the control solution and evaluated over 30 days. Clinical efficacy was evaluated by clinical signs and cytological organism counts. In vitro antimicrobial susceptibility was evaluated by the minimum inhibitory concentration (MIC). After treatment with the test solution, clinical signs continuously decreased and cytological scores were significantly reduced. The results of MIC testing showed that the test solution was potent against the common pathogenic causes of canine OE. In this study, the most common causative pathogens were Staphylococcus pseudintermedius, Pseudomonas spp. and Malassezia pachydermatis. No issues related to safety were identified. Based on these results, this new ototopical drug can be used as first line treatment for canine OE.

Toxicogenomics and Cell-based Assays for Toxicology

  • Tong, Weida;Fang, Hong;Mendrick, Donna
    • Interdisciplinary Bio Central
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    • v.1 no.3
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    • pp.10.1-10.5
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    • 2009
  • Toxicity is usually investigated using a set of standardized animal-based studies which, unfortunately, fail to detect all compounds that induce human adverse events and do not provide detailed mechanistic information of observed toxicity. As an alternative to conventional toxicology, toxicogenomics takes advantage of currently advanced technologies in genomics, proteomics, metabolomics, and bioinformatics to gain a molecular level understanding of toxicity and to enhance the predictive power of toxicity testing in drug development and risk/safety assessment. In addition, there has been a renewed interest, particularly in various government agencies, to prioritize and/or supplement animal testing with a battery of mechanistically informative in vitro assays. This article provides a brief summary of the issues, challenges and lessons learned in these fields and discuss the ways forward to further advance toxicology using these technologies.

Assessment of the Dermal and Ocular Irritation Potential of Lomefloxacin by Using In Vitro Methods

  • Ahn, Jun-Ho;Eum, Ki-Hwan;Lee, Mi-Chael
    • Toxicological Research
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    • v.26 no.1
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    • pp.9-14
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    • 2010
  • The evaluation of eye and skin irritation potential is essential to ensuring the safety of human in contact with a wide variety of substances. Despite this importance of irritation test, little is known with respect to the irritation potency of lomefloxacin, a fluoroquinolone antibiotic, which has been known to cause phototoxicity with an abnormal reaction of the skin. Thus, to investigate the tendency of lomefloxacin to cause eye and skin irritation, we carried out in vitro eye irritation test using Balb/c 3T3, and in vitro skin irritation test using $KeraSkin^{TM}$ human skin model system. 3T3 neutral red uptake assay has been proposed as a potential replacement alternative for the Draize Eye irritation test. In this study, the $IC_{50}$ value obtained for lomefloxacin was 375 ${\mu}g$. According to the classification model used for determining in vitro categories, lomefloxacin was classified as moderately irritant. For evaluation of skin irritation, engineered epidermal equivalents ($KeraSkin^{TM}$) were subjected to 10 and 25 mg of lomefloxacin for 15 minutes. Tissue damage was assessed by tissue viability evaluation, and by the release of a pro-inflammatory mediator, interleukin- 1${\alpha}$. Lomefloxacin increased the interleukin-1${\alpha}$ release after 15 minutes of exposure and 42 hours of post incubation, although no decrease in viability was observed. Therefore, lomefloxacin is considered to be moderately irritant to skin and eye.

Alternative Methods for Testing Botulinum Toxin: Current Status and Future Perspectives

  • Nepal, Mahesh Raj;Jeong, Tae Cheon
    • Biomolecules & Therapeutics
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    • v.28 no.4
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    • pp.302-310
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    • 2020
  • Botulinum toxins are neurotoxic modular proteins composed of a heavy chain and a light chain connected by a disulfide bond and are produced by Clostridium botulinum. Although lethally toxic, botulinum toxin in low doses is clinically effective in numerous medical conditions, including muscle spasticity, strabismus, hyperactive urinary bladder, excessive sweating, and migraine. Globally, several companies are now producing products containing botulinum toxin for medical and cosmetic purposes, including the reduction of facial wrinkles. To test the efficacy and toxicity of botulinum toxin, animal tests have been solely and widely used, resulting in the inevitable sacrifice of hundreds of animals. Hence, alternative methods are urgently required to replace animals in botulinum toxin testing. Here, the various alternative methods developed to test the toxicity and efficacy of botulinum toxins have been briefly reviewed and future perspectives have been detailed.

Antimicrobial Activity and Chemical Composition of Some Essential Oils

  • Arldogan, Buket-Cicioglu;Baydar, Hasan;Kaya, Selcuk;Demirci, Mustafa;Ozbasar, Demir;Mumcu, Ethem
    • Archives of Pharmacal Research
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    • v.25 no.6
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    • pp.860-864
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    • 2002
  • In this study the composition and antimicrobial properties of essential oils obtained from Origanum onites, Mentha piperita, Juniperus exalsa, Chrysanthemum indicum, Lavandula hybrida, Rosa damascena, Echinophora tenuifolia, Foeniculum vulgare were examined. To evaluate the in vitro antibacterial activities of these eight aromatic extracts; their in vitro antimicrobial activities were determined by disk diffusion testing, according to the NCCLS criteria. Escherichia coli (ATTC 25922), Staphylococcus aureus (ATCC 25923) and Pseudomonas aeruginosa (ATTC 27853 were used as standard test bacterial strains. Origanum onites recorded antimicrobial activity against all test bacteria, and was strongest against Staphylococcus aureus. For Rosa damascena, Mentha piperita and Lavandula hybrida antimicrobial activity was recorded only to Staphylococcus aureus. Juniperus exalsa, and Chrysanthemum indicum exhibited antibacterial activities against both Staphylococcus aureus and Escherichia coli. We also examined the in vitro artimicrobial activities of some components of the essential oils and found some components with antimicrobial activity.

Guidelines for Manufacturing and Application of Organoids: Kidney

  • Hyun Mi Kang;Dong Sung Kim;Yong Kyun Kim;Kunyoo Shin;Sun-Ju Ahn;Cho-Rok Jung
    • International Journal of Stem Cells
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    • v.17 no.2
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    • pp.141-146
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    • 2024
  • Recent advancements in organoid technology have led to a vigorous movement towards utilizing it as a substitute for animal experiments. Organoid technology offers versatile applications, particularly in toxicity testing of pharmaceuticals or chemical substances. However, for the practical use in toxicity testing, minimal guidance is required to ensure reliability and relevance. This paper aims to provide minimal guidelines for practical uses of kidney organoids derived from human pluripotent stem cells as a toxicity evaluation model in vitro.