WorldWideScience

Sample records for vitro test systems

  1. Validation of artificial skin equivalents as in vitro testing systems

    Science.gov (United States)

    Schmitt, Robert; Marx, Ulrich; Walles, Heike; Schober, Lena

    2011-03-01

    With the increasing complexity of the chemical composition of pharmaceuticals, cosmetics and everyday substances, the awareness of potential health issues and long term damages for humanoid organs is shifting into focus. Artificial in vitro testing systems play an important role in providing reliable test conditions and replacing precarious animal testing. Especially artificial skin equivalents ASEs are used for a broad spectrum of studies like penetration, irritation and corrosion of substances. One major challenge in tissue engineering is the qualification of each individual ASE as in vitro testing system. Due to biological fluctuations, the stratum corneum hornified layer of some ASEs may not fully develop or other defects might occur. For monitoring these effects we developed an fully automated Optical Coherence Tomography device. Here, we present different methods to characterize and evaluate the quality of the ASEs based on image and data processing of OCT B-scans. By analysing the surface structure, defects, like cuts or tears, are detectable. A further indicator for the quality of the ASE is the morphology of the tissue. This allows to determine if the skin model has reached the final growth state. We found, that OCT is a well suited technology for automatically characterizing artificial skin equivalents and validating the application as testing system.

  2. Accelerated in-vitro release testing methods for extended-release parenteral dosage forms.

    Science.gov (United States)

    Shen, Jie; Burgess, Diane J

    2012-07-01

    This review highlights current methods and strategies for accelerated in-vitro drug release testing of extended-release parenteral dosage forms such as polymeric microparticulate systems, lipid microparticulate systems, in-situ depot-forming systems and implants. Extended-release parenteral dosage forms are typically designed to maintain the effective drug concentration over periods of weeks, months or even years. Consequently, 'real-time' in-vitro release tests for these dosage forms are often run over a long time period. Accelerated in-vitro release methods can provide rapid evaluation and therefore are desirable for quality control purposes. To this end, different accelerated in-vitro release methods using United States Pharmacopeia (USP) apparatus have been developed. Different mechanisms of accelerating drug release from extended-release parenteral dosage forms, along with the accelerated in-vitro release testing methods currently employed are discussed. Accelerated in-vitro release testing methods with good discriminatory ability are critical for quality control of extended-release parenteral products. Methods that can be used in the development of in-vitro-in-vivo correlation (IVIVC) are desirable; however, for complex parenteral products this may not always be achievable. © 2012 The Authors. JPP © 2012 Royal Pharmaceutical Society.

  3. Accelerated in vitro release testing methods for extended release parenteral dosage forms

    Science.gov (United States)

    Shen, Jie; Burgess, Diane J.

    2012-01-01

    Objectives This review highlights current methods and strategies for accelerated in vitro drug release testing of extended release parenteral dosage forms such as polymeric microparticulate systems, lipid microparticulate systems, in situ depot-forming systems, and implants. Key findings Extended release parenteral dosage forms are typically designed to maintain the effective drug concentration over periods of weeks, months or even years. Consequently, “real-time” in vitro release tests for these dosage forms are often run over a long time period. Accelerated in vitro release methods can provide rapid evaluation and therefore are desirable for quality control purposes. To this end, different accelerated in vitro release methods using United States Pharmacopoeia (USP) apparatus have been developed. Different mechanisms of accelerating drug release from extended release parenteral dosage forms, along with the accelerated in vitro release testing methods currently employed are discussed. Conclusions Accelerated in vitro release testing methods with good discriminatory ability are critical for quality control of extended release parenteral products. Methods that can be used in the development of in vitro-in vivo correlation (IVIVC) are desirable, however for complex parenteral products this may not always be achievable. PMID:22686344

  4. In Vitro Toxicity testing in the 21st Century

    Directory of Open Access Journals (Sweden)

    Erwin L Roggen

    2011-02-01

    Full Text Available The National Research Council (NRC article Toxicity Testing in the 21st Century: A vision and A Strategy (National Research Council, 2007 was written to bring attention to the application of scientific advances for use in toxicity tests so that chemicals can be tested in a more time and cost efficient manner while providing a more relevant and mechanistic insight into the toxic potential of a compound.Development of tools for in vitro toxicity testing constitutes an important activity of this vision and contributes to the provision of test systems as well as data that are essential for the development of computer modelling tools for e.g. system biology, physiologically-based modelling. This article intends to highlight some of the issues that have to be addressed in order to make in vitro toxicity testing a reality in the 21st century.

  5. Physiological parameters for oral delivery and in vitro testing.

    Science.gov (United States)

    Mudie, Deanna M; Amidon, Gordon L; Amidon, Gregory E

    2010-10-04

    Pharmaceutical solid oral dosage forms must undergo dissolution in the intestinal fluids of the gastrointestinal tract before they can be absorbed and reach the systemic circulation. Therefore, dissolution is a critical part of the drug-delivery process. The rate and extent of drug dissolution and absorption depend on the characteristics of the active ingredient as well as properties of the dosage form. Just as importantly, characteristics of the physiological environment such as buffer species, pH, bile salts, gastric emptying rate, intestinal motility, and hydrodynamics can significantly impact dissolution and absorption. While significant progress has been made since 1970 when the first compendial dissolution test was introduced (USP apparatus 1), current dissolution testing does not take full advantage of the extensive physiologic information that is available. For quality control purposes, where the question is one of lot-to-lot consistency in performance, using nonphysiologic test conditions that match drug and dosage form properties with practical dissolution media and apparatus may be appropriate. However, where in vitro-in vivo correlations are desired, it is logical to consider and utilize knowledge of the in vivo condition. This publication critically reviews the literature that is relevant to oral human drug delivery. Physiologically relevant information must serve as a basis for the design of dissolution test methods and systems that are more representative of the human condition. As in vitro methods advance in their physiological relevance, better in vitro-in vivo correlations will be possible. This will, in turn, lead to in vitro systems that can be utilized to more effectively design dosage forms that have improved and more consistent oral bioperformance.

  6. Implementation of in vitro replacement technologies in regulatory drug testing - An innovation systems perspective

    NARCIS (Netherlands)

    Kooijman, M.; Van Meer, P.J.K.; Moors, E.H.M.; Hekkert, M.P.; Schellekens, H.

    2011-01-01

    The replacement of in vivo methods by in vitro methods in regulatory drug testing is rare. The aim of this research is to identify barriers and drivers of the replacement of in vivo methods by in vitro methods in Europe. We studied two cases. The first case is the Draize eye test. Since 2009, the in

  7. Potential countersample materials for in vitro simulation wear testing.

    Science.gov (United States)

    Shortall, Adrian C; Hu, Xiao Q; Marquis, Peter M

    2002-05-01

    Any laboratory investigation of the wear resistance of dental materials needs to consider oral conditions so that in vitro wear results can be correlated with in vivo findings. The choice of the countersample is a critical factor in establishing the pattern of tribological wear and in achieving an efficient in vitro wear testing system. This research investigated the wear behavior and surface characteristics associated with three candidate countersample materials used for in vitro wear testing in order to identify a possible suitable substitute for human dental enamel. Three candidate materials, stainless steel, steatite and dental porcelain were evaluated and compared to human enamel. A variety of factors including hardness, wear surface evolution and frictional coefficients were considered, relative to the tribology of the in vivo situation. The results suggested that the dental porcelain investigated bore the closest similarity to human enamel of the materials investigated. Assessment of potential countersample materials should be based on the essential tribological simulation supported by investigations of mechanical, chemical and structural properties. The selected dental porcelain had the best simulating ability among the three selected countersample materials and this class of material may be considered as a possible countersample material for in vitro wear test purposes. Further studies are required, employing a wider range of dental ceramics, in order to optimise the choice of countersample material for standardized in vitro wear testing.

  8. Toxicity testing: the search for an in vitro alternative to animal testing.

    Science.gov (United States)

    May, J E; Xu, J; Morse, H R; Avent, N D; Donaldson, C

    2009-01-01

    Prior to introduction to the clinic, pharmaceuticals must undergo rigorous toxicity testing to ensure their safety. Traditionally, this has been achieved using in vivo animal models. However, besides ethical reasons, there is a continual drive to reduce the number of animals used for this purpose due to concerns such as the lack of concordance seen between animal models and toxic effects in humans. Adequate testing to ensure any toxic metabolites are detected can be further complicated if the agent is administered in a prodrug form, requiring a source of cytochrome P450 enzymes for metabolism. A number of sources of metabolic enzymes have been utilised in in vitro models, including cell lines, primary human tissue and liver extracts such as S9. This review examines current and new in vitro models for toxicity testing, including a new model developed within the authors' laboratory utilising HepG2 liver spheroids within a co-culture system to examine the effects of chemotherapeutic agents on other cell types.

  9. Surface modification of nano-silica on the ligament advanced reinforcement system for accelerated bone formation: primary human osteoblasts testing in vitro and animal testing in vivo.

    Science.gov (United States)

    Li, Mengmeng; Wang, Shiwen; Jiang, Jia; Sun, Jiashu; Li, Yuzhuo; Huang, Deyong; Long, Yun-Ze; Zheng, Wenfu; Chen, Shiyi; Jiang, Xingyu

    2015-05-07

    The Ligament Advanced Reinforcement System (LARS) has been considered as a promising graft for ligament reconstruction. To improve its biocompatibility and effectiveness on new bone formation, we modified the surface of a polyethylene terephthalate (PET) ligament with nanoscale silica using atom transfer radical polymerization (ATRP) and silica polymerization. The modified ligament is tested by both in vitro and in vivo experiments. Human osteoblast testing in vitro exhibits an ∼21% higher value in cell viability for silica-modified grafts compared with original grafts. Animal testing in vivo shows that there is new formed bone in the case of a nanoscale silica-coated ligament. These results demonstrate that our approach for nanoscale silica surface modification on LARS could be potentially applied for ligament reconstruction.

  10. The current limitations of in vitro genotoxicity testing and their relevance to the in vivo situation.

    Science.gov (United States)

    Nesslany, Fabrice

    2017-08-01

    The standard regulatory core battery of genotoxicity tests generally includes 2 or 3 validated tests with at least one in vitro test in bacteria and one in vitro test on cell cultures. However, limitations in in vitro genotoxicity testing may exist at many levels. The knowledge of the underlying mechanisms of genotoxicity is particularly useful to assess the level of relevance for the in vivo situation. In order to avoid wrong conclusions regarding the actual genotoxicity status of any test substance, it appears very important to be aware of the various origins of related bias leading to 'false positives and negatives' by using in vitro methods. Among these, mention may be made on the metabolic activation system, experimental (extreme) conditions, specificities of the test systems implemented, cell type used etc. The knowledge of the actual 'limits' of the in vitro test systems used is clearly an advantage and may contribute to avoid some pitfalls in order to better assess the level of relevance for the in vivo situation. Copyright © 2016. Published by Elsevier Ltd.

  11. RNA biology in a test tube--an overview of in vitro systems/assays.

    Science.gov (United States)

    Roca, Xavier; Karginov, Fedor V

    2012-01-01

    In vitro systems have provided a wealth of information in the field of RNA biology, as they constitute a superior and sometimes the unique approach to address many important questions. Such cell-free methods can be sorted by the degree of complexity of the preparation of enzymatic and/or regulatory activity. Progress in the study of pre-mRNA processing has largely relied on traditional in vitro methods, as these reactions have been recapitulated in cell-free systems. The pre-mRNA capping, editing, and cleavage/polyadenylation reactions have even been reconstituted using purified components, and the enzymes responsible for catalysis have been characterized by such techniques. In vitro splicing using nuclear or cytoplasmic extracts has yielded clues on spliceosome assembly, kinetics, and mechanisms of splicing and has been essential to elucidate the function of splicing factors. Coupled systems have been important to functionally connect distinct processes, like transcription and splicing. Extract preparation has also been adapted to cells from a variety of tissues and species, revealing general versus species-specific mechanisms. Cell-free assays have also been applied to newly discovered pathways such as those involving small RNAs, including microRNAs (miRNAs), small interfering RNAs (siRNAs), and Piwi-interacting RNAs (piRNAs). The first two pathways have been well characterized largely by in vitro methods, which need to be developed for piRNAs. Finally, new techniques, such as single-molecule studies, are continuously being established, providing new and important insights into the field. Thus, in vitro approaches have been, are, and will continue being at the forefront of RNA research. Copyright © 2012 John Wiley & Sons, Ltd.

  12. Toward the establishment of standardized in vitro tests for lipid-based formulations, part 4

    DEFF Research Database (Denmark)

    Williams, Hywel D; Sassene, Philip; Kleberg, Karen

    2014-01-01

    The Lipid Formulation Classification System Consortium looks to develop standardized in vitro tests and to generate much-needed performance criteria for lipid-based formulations (LBFs). This article highlights the value of performing a second, more stressful digestion test to identify LBFs near...... a performance threshold and to facilitate lead formulation selection in instances where several LBF prototypes perform adequately under standard digestion conditions (but where further discrimination is necessary). Stressed digestion tests can be designed based on an understanding of the factors that affect LBF...... development, and facilitate dialogue with the regulatory authorities. This classification system is based on the concept that performance evaluations across three in vitro tests, designed to subject a LBF to progressively more challenging conditions, will enable effective LBF discrimination and performance...

  13. Evaluation of seven in vitro alternatives for ocular safety testing.

    Science.gov (United States)

    Bruner, L H; Kain, D J; Roberts, D A; Parker, R D

    1991-07-01

    Seven in vitro assays were evaluated to determine if any were useful as screening procedures in ocular safety assessment. Seventeen test materials (chemicals, household cleaners, hand soaps, dishwashing liquids, shampoos, and liquid laundry detergents) were tested in each assay. In vivo ocular irritation scores for the materials were obtained from existing rabbit low volume eye test (LVET) data. The seven assays evaluated included the silicon microphysiometer (SM), luminescent bacteria toxicity test (LBT), neutral red assay (NR), total protein assay (TP), Tetrahymena thermophila motility assay (TTMA), bovine eye/chorioallantoic membrane assay (BE/CAM), and the EYTEX system (ETS). For the seventeen materials used in this study there was a significant correlation between the in vivo irritant potential and in vitro data for all the tests except the EYTEX System (SM, r = -0.87; LBT, r = -0.91; NR, r = -0.85; TTMA, r = 0.78; TP, r = -0.86; ETS, r = 0.29). The irritation classifications provided by the BE/CAM also did not correspond with the actual in vivo irritancy potential of the test materials. The result of this study suggested it may be possible to classify materials into broad irritancy categories with some of the assays. This would allow their use as screens prior to limited in vivo confirmation in the ocular safety assessment process.

  14. Sex in a test tube: testing the benefits of in vitro recombination.

    Science.gov (United States)

    Pesce, Diego; Lehman, Niles; de Visser, J Arjan G M

    2016-10-19

    The origin and evolution of sex, and the associated role of recombination, present a major problem in biology. Sex typically involves recombination of closely related DNA or RNA sequences, which is fundamentally a random process that creates but also breaks up beneficial allele combinations. Directed evolution experiments, which combine in vitro mutation and recombination protocols with in vitro or in vivo selection, have proved to be an effective approach for improving functionality of nucleic acids and enzymes. As this approach allows extreme control over evolutionary conditions and parameters, it also facilitates the detection of small or position-specific recombination benefits and benefits associated with recombination between highly divergent genotypes. Yet, in vitro approaches have been largely exploratory and motivated by obtaining improved end products rather than testing hypotheses of recombination benefits. Here, we review the various experimental systems and approaches used by in vitro studies of recombination, discuss what they say about the evolutionary role of recombination, and sketch their potential for addressing extant questions about the evolutionary role of sex and recombination, in particular on complex fitness landscapes. We also review recent insights into the role of 'extracellular recombination' during the origin of life.This article is part of the themed issue 'Weird sex: the underappreciated diversity of sexual reproduction'. © 2016 The Author(s).

  15. In vitro pyrogen test for toxic or immunomodulatory drugs

    OpenAIRE

    Daneshian, Mardas; Guenther, Armin; Wendel, Albrecht; Hartung, Thomas; Aulock, Sonja von

    2006-01-01

    Pyrogenic contaminations of some classes of injectable drugs, e.g. toxic or immunomodulatory as well as false-positive drugs, represent a major risk which cannot yet be excluded due to the limitations of current tests. Here we describe a modification of the In vitro Pyrogen Test termed AWIPT (Adsorb, Wash, In vitro Pyrogen Test), which addresses this problem by introducing a pre-incubation step in which pyrogenic contaminations in the test sample are adsorbed to albumin-coated beads. After ri...

  16. Toward the establishment of standardized in vitro tests for lipid-based formulations, part 4: proposing a new lipid formulation performance classification system.

    Science.gov (United States)

    Williams, Hywel D; Sassene, Philip; Kleberg, Karen; Calderone, Marilyn; Igonin, Annabel; Jule, Eduardo; Vertommen, Jan; Blundell, Ross; Benameur, Hassan; Müllertz, Anette; Porter, Christopher J H; Pouton, Colin W

    2014-08-01

    The Lipid Formulation Classification System Consortium looks to develop standardized in vitro tests and to generate much-needed performance criteria for lipid-based formulations (LBFs). This article highlights the value of performing a second, more stressful digestion test to identify LBFs near a performance threshold and to facilitate lead formulation selection in instances where several LBF prototypes perform adequately under standard digestion conditions (but where further discrimination is necessary). Stressed digestion tests can be designed based on an understanding of the factors that affect LBF performance, including the degree of supersaturation generated on dispersion/digestion. Stresses evaluated included decreasing LBF concentration (↓LBF), increasing bile salt, and decreasing pH. Their capacity to stress LBFs was dependent on LBF composition and drug type: ↓LBF was a stressor to medium-chain glyceride-rich LBFs, but not more hydrophilic surfactant-rich LBFs, whereas decreasing pH stressed tolfenamic acid LBFs, but not fenofibrate LBFs. Lastly, a new Performance Classification System, that is, LBF composition independent, is proposed to promote standardized LBF comparisons, encourage robust LBF development, and facilitate dialogue with the regulatory authorities. This classification system is based on the concept that performance evaluations across three in vitro tests, designed to subject a LBF to progressively more challenging conditions, will enable effective LBF discrimination and performance grading. © 2014 Wiley Periodicals, Inc. and the American Pharmacists Association.

  17. In vitro tests and ethnopharmacological investigations: wound healing as an example.

    Science.gov (United States)

    Houghton, P J; Hylands, P J; Mensah, A Y; Hensel, A; Deters, A M

    2005-08-22

    In vitro tests are now widely employed in ethnopharmacological research because of ethical reasons and their usefulness in bioactive-guided fractionation and determination of active compounds. For many disease conditions, a variety of in vitro tests can now be employed as the biochemical mechanisms underlying disease and healing processes are understood. Approaches to the in vitro investigations of wound healing processes are exemplified by studies on extracts of Buddleja species and three Ghanaian species Spathodea campanulata, Commelina diffusa and Secamone afzelii. Most studies have been carried out on Buddleja officinalis or Buddleja globosa. The extracts have been shown to have anti-inflammatory and antioxidant properties due to flavonoids, triterpenoids, diterpenoids and caffeic acid derivatives. There appears to a slight effect on proliferation of fibroblasts at lower concentrations, but this was not significant, and higher concentrations appeared to be cytotoxic. Novel findings are the ability of Buddleja globosa leaf extracts to induce differentiation in keratinocytes and to alter the profile of proteins produced by cultured fibroblasts. Extracts also had some effect on lattice contraction. The three Ghanaian species examined show a mixture of antimicrobial and antioxidant activities. The evolution over recent years of tests for wound healing, from in vivo tests to cell-based systems and chemical reactions and on to investigations into effects on secondary messengers and protein expression, is described.

  18. In vitro pyrogen test for toxic or immunomodulatory drugs.

    Science.gov (United States)

    Daneshian, Mardas; Guenther, Armin; Wendel, Albrecht; Hartung, Thomas; von Aulock, Sonja

    2006-06-30

    Pyrogenic contaminations of some classes of injectable drugs, e.g. toxic or immunomodulatory as well as false-positive drugs, represent a major risk which cannot yet be excluded due to the limitations of current tests. Here we describe a modification of the In vitro Pyrogen Test termed AWIPT (Adsorb, Wash, In vitro Pyrogen Test), which addresses this problem by introducing a pre-incubation step in which pyrogenic contaminations in the test sample are adsorbed to albumin-coated beads. After rinsing, the beads are incubated with human whole blood and the release of the endogenous pyrogen interleukin-1beta is measured as a marker of pyrogenic activity. Intentional contaminations with lipopolysaccharide were retrieved from the chemotherapeutic agents paclitaxel, cisplatin and liposomal daunorubicin, the antibiotic gentamicin, the antifungal agent liposomal amphotericin B, and the corticosteroid prednisolone at lower dilutions than in the standard in vitro pyrogen test. This represents a promising new approach for the detection of pyrogenic contamination in drugs or in drugs containing interfering additives and should lead to improved safety levels.

  19. The development of in vitro mutagenicity testing systems using T-lymphocytes

    International Nuclear Information System (INIS)

    Albertini, R.J.

    1992-05-01

    This work has focused on the development of in vitro T-cell mutation assays. Conditions have been defined to measure the in vitro induction of mutations at the hypoxanthine-guanine phosphoribosyl transferase (hprt) locus in human T-lymphocytes. This assay is a parallel to our in vivo hprt assay, in that the same cells are utilized. However, the in vitro assay allows for carefully controlled dose response studies. 21 refs., 16 figs., 13 tabs

  20. In vitro test for pyrogenes in radiopharmaceuticals

    Energy Technology Data Exchange (ETDEWEB)

    Jovanovic, V; Zmbova, B; Bzenic, J [Institut za Nuklearne Nauke Boris Kidric, Belgrade (Yugoslavia); Berkes, J [Institut za Biohemije, Belgrade (Yugoslavia)

    1978-05-01

    Procedure and results of determination of pyrogenic substances in radiopharmaceutical preparations by an in vitro method based on the reaction between bacterial endotoxine and Limulus Amebocyte Lysate are presented. The advantage of this method as compared to the test in experimental animals performed so far has also been analyzed and proved by the fact that it enables avoidance of introduction of radioactive materials in experimental animals and of radiation effects on the results obtained in efficiency studies. The in vitro method is a quick one and requires only small quantities of the radiopharmaceutical preparation to be examined.

  1. In vitro metabolism and bioavailability tests for endocrine active substances: What is needed next for regulatory purposes?

    Science.gov (United States)

    Legistation and prospective legislative proposals internationally (may) require that chemicals be tested for their ability to disrupt the hormonal systems of mammals. Chemicals found to test positive in vitro are considered to be endocrine active substances (EAS) and may be puta...

  2. Interpreting in vitro developmental toxicity test battery results: The consideration of toxicokinetics

    NARCIS (Netherlands)

    Bosgra, S.; Westerhout, J.

    2015-01-01

    In the EU collaborative project ChemScreen an alternative, in vitro assay-based test strategy was developed to screen compounds for reproductive toxicity. A toxicokinetic modeling approach was used to allow quantitative comparison between effective concentrations in the in vitro test battery and

  3. Comparing in vitro activity of tigecycline by using the disk diffusion test, the manual microdilution method, and the VITEK 2 automated system.

    Science.gov (United States)

    Leal Castro, A L; Buitrago Gutierrez, G; Ovalle, V; Cortes, J A; Alvarez, C A

    2010-01-01

    Tigecycline is a broad spectrum antibiotic having activity against multiresistant isolates. In vitro susceptibility testing is difficult to perform with the use of traditional microbiological techniques. The aim of this study was to evaluate the disk diffusion test with three different Mueller-Hinton agar brands, and the Vitek 2 automated system in comparison with the standard broth microdilution method against 200 gram-negative isolates (Escherichia coil, Klebsiella pneumoniae, Enterobacter cloacae, Serratia marcescens and Acinetobacter baumannii). Among Enterobacteriaceae, the Becton Dickinson agar had the lowest rate of minor (32.5%) and major errors (3.8%). No very major errors were found. For A. baumanni, the rate of minor and major errors was lower. A high rate of agreement (94%) was found between the broth microdilution method and the Vitek 2 system. Our results show that there are important differences between agars used for the disk diffusion test, and that Vitek 2 is a valid tool for susceptibility testing in clinical laboratories.

  4. In Vitro Tests for Aerosol Deposition. V: Using Realistic Testing to Estimate Variations in Aerosol Properties at the Trachea.

    Science.gov (United States)

    Wei, Xiangyin; Hindle, Michael; Delvadia, Renishkumar R; Byron, Peter R

    2017-10-01

    The dose and aerodynamic particle size distribution (APSD) of drug aerosols' exiting models of the mouth and throat (MT) during a realistic inhalation profile (IP) may be estimated in vitro and designated Total Lung Dose, TLD in vitro , and APSD TLDin vitro , respectively. These aerosol characteristics likely define the drug's regional distribution in the lung. A general method was evaluated to enable the simultaneous determination of TLD in vitro and APSD TLDin vitro for budesonide aerosols' exiting small, medium and large VCU-MT models. Following calibration of the modified next generation pharmaceutical impactor (NGI) at 140 L/min, variations in aerosol dose and size exiting MT were determined from Budelin ® Novolizer ® across the IPs reported by Newman et al., who assessed drug deposition from this inhaler by scintigraphy. Values for TLD in vitro from the test inhaler determined by the general method were found to be statistically comparable to those using a filter capture method. Using new stage cutoffs determined by calibration of the modified NGI at 140 L/min, APSD TLDin vitro profiles and mass median aerodynamic diameters at the MT exit (MMAD TLDin vitro ) were determined as functions of MT geometric size across Newman's IPs. The range of mean values (n ≥ 5) for TLD in vitro and MMAD TLDin vitro for this inhaler extended from 6.2 to 103.0 μg (3.1%-51.5% of label claim) and from 1.7 to 3.6 μm, respectively. The method enables reliable determination of TLD in vitro and APSD TLDin vitro for aerosols likely to enter the trachea of test subjects in the clinic. By simulating realistic IPs and testing in different MT models, the effects of major variables on TLD in vitro and APSD TLDin vitro may be studied using the general method described in this study.

  5. In Vitro Testing of Scaffolds for Mesenchymal Stem Cell-Based Meniscus Tissue Engineering—Introducing a New Biocompatibility Scoring System

    Directory of Open Access Journals (Sweden)

    Felix P. Achatz

    2016-04-01

    Full Text Available A combination of mesenchymal stem cells (MSCs and scaffolds seems to be a promising approach for meniscus repair. To facilitate the search for an appropriate scaffold material a reliable and objective in vitro testing system is essential. This paper introduces a new scoring for this purpose and analyzes a hyaluronic acid (HA gelatin composite scaffold and a polyurethane scaffold in combination with MSCs for tissue engineering of meniscus. The pore quality and interconnectivity of pores of a HA gelatin composite scaffold and a polyurethane scaffold were analyzed by surface photography and Berliner-Blau-BSA-solution vacuum filling. Further the two scaffold materials were vacuum-filled with human MSCs and analyzed by histology and immunohistochemistry after 21 days in chondrogenic media to determine cell distribution and cell survival as well as proteoglycan production, collagen type I and II content. The polyurethane scaffold showed better results than the hyaluronic acid gelatin composite scaffold, with signs of central necrosis in the HA gelatin composite scaffolds. The polyurethane scaffold showed good porosity, excellent pore interconnectivity, good cell distribution and cell survival, as well as an extensive content of proteoglycans and collagen type II. The polyurethane scaffold seems to be a promising biomaterial for a mesenchymal stem cell-based tissue engineering approach for meniscal repair. The new score could be applied as a new standard for in vitro scaffold testing.

  6. In vitro vaccine potency testing: a proposal for reducing animal use for requalification testing.

    Science.gov (United States)

    Brown, K; Stokes, W

    2012-01-01

    This paper proposes a program under which the use of animals for requalification of in vitro potency tests could be eliminated. Standard References (USDA/CVB nomenclature) would be developed, characterized, stored and monitored by selected reference laboratories worldwide. These laboratories would employ scientists skilled in protein and glycoprotein chemistry and equipped with state-of-the-art instruments for required analyses. After Standard References are established, the reference laboratories would provide them to the animal health industry as "gold standards". Companies would then establish and validate a correlation between the Standard Reference and the company Master Reference (USDA/CVB nomenclature) using an internal in vitro assay. After this correlation is established, the company could use the Standard References for qualifying, monitoring and requalifying company Master References without the use of animals. Such a program would eliminate the need for animals for requalification of Master References and the need for each company to develop and validate a battery of Master Reference Monitoring assays. It would also provide advantages in terms of reduced costs and reduced time for requalification testing. As such it would provide a strong incentive for companies to develop and use in vitro assays for potency testing.

  7. Use of external metabolizing systems when testing for endocrine disruption in the T-screen assay

    International Nuclear Information System (INIS)

    Taxvig, Camilla; Olesen, Pelle Thonning; Nellemann, Christine

    2011-01-01

    Although, it is well-established that information on the metabolism of a substance is important in the evaluation of its toxic potential, there is limited experience with incorporating metabolic aspects into in vitro tests for endocrine disrupters. The aim of the current study was a) to study different in vitro systems for biotransformation of ten known endocrine disrupting chemicals (EDs): five azole fungicides, three parabens and 2 phthalates, b) to determine possible changes in the ability of the EDs to bind and activate the thyroid receptor (TR) in the in vitro T-screen assay after biotransformation and c) to investigate the endogenous metabolic capacity of the GH3 cells, the cell line used in the T-screen assay, which is a proliferation assay used for the in vitro detection of agonistic and antagonistic properties of compounds at the level of the TR. The two in vitro metabolizing systems tested the human liver S9 mix and the PCB-induced rat microsomes gave an almost complete metabolic transformation of the tested parabens and phthalates. No marked difference the effects in the T-screen assay was observed between the parent compounds and the effects of the tested metabolic extracts. The GH3 cells themselves significantly metabolized the two tested phthalates dimethyl phthalate (DMP) and diethyl phthalate (DEP). Overall the results and qualitative data from the current study show that an in vitro metabolizing system using liver S9 or microsomes could be a convenient method for the incorporation of metabolic and toxicokinetic aspects into in vitro testing for endocrine disrupting effects.

  8. Vancomycin-resistant enterococci: validation of susceptibility testing and in vitro activity of novel antibiotics

    DEFF Research Database (Denmark)

    Rathe, Mathias; Lise, Kristensen,; Ellermann-Eriksen, Svend

    Vancomycin-resistant enterococci: validation of susceptibility testing and in vitro activity of novel antibiotics......Vancomycin-resistant enterococci: validation of susceptibility testing and in vitro activity of novel antibiotics...

  9. Chemosensitivity and radiosensitivity testing of freshly explanted human tumour cells in vitro

    International Nuclear Information System (INIS)

    Wells, J.

    1977-10-01

    In this thesis, in vitro testing for the chemosensitivity and radiosensitivity of freshly explanted human tumour cells is described. The cells were incubated with anti-tumour drugs and either a 6-day growth test performed or a clonal growth test as a measure of survival of cell reproductive capacity. It was shown that if one aims to develop a suitable in vitro method for predicting the subsequent response of human tumour cells in situ to cytotoxic chemotherapy, the test procedure must be initiated before the explanted cells have undergone significant growth in vitro. The survival of the reproductive capacity of tumour cell explants following X-radiation was also studied. Using a 'feeder' layer technique, values for the survival curve parameter Dsub(q) were in the range 400-610 rad and the values for D 0 were in the range 120-160 rad. The shape of the X-ray survival curves did not change when cells were retested after repeated subculturing in vitro. Therefore, unlike chemosensitivity measured by the same biological end-point, radiosensitivity apparently does not change once cells have reached their maximum growth potential. (UK)

  10. Correlation between the results of in vitro and in vivo chromosomal damage tests in consideration of exposure levels of test chemicals.

    Science.gov (United States)

    Yamamura, Eiji; Aruga, Chinami; Muto, Shigeharu; Baba, Nobuyuki; Uno, Yoshifumi

    2018-01-01

    We examined the correlation between the results of in vitro and in vivo chromosomal damage tests by using in-house data of 18 pharmaceutical candidates that showed positive results in the in vitro chromosomal aberration or micronucleus test using CHL/IU cells, and quantitatively analyzed them especially in regard to exposure levels of the compounds. Eight compounds showed that the exposure levels [maximum plasma concentration (C max ) and AUC 0-24h ] were comparable with or higher than the in vitro exposure levels [the lowest effective (positive) concentration (LEC) and AUC vitro  = LEC (μg/mL) × treatment time (h)]. Among them, 3 compounds were positive in the in vivo rodent micronucleus assays using bone marrow cells. For 2 compounds, cytotoxicity might produce false-positive results in the in vitro tests. One compound showed in vitro positive results only in the condition with S9 mix which indicated sufficient concentration of unidentified active metabolite(s) might not reach the bone marrow to induce micronuclei. These facts suggested that the in vivo exposure levels being equal to or higher than the in vitro exposure levels might be an important factor to detect in vivo chromosomal damage induced by test chemicals.

  11. Transgenic Mouse Models Transferred into the Test Tube: New Perspectives for Developmental Toxicity Testing In Vitro?

    Science.gov (United States)

    Kugler, Josephine; Luch, Andreas; Oelgeschläger, Michael

    2016-10-01

    Despite our increasing understanding of molecular mechanisms controlling embryogenesis, the identification and characterization of teratogenic substances still heavily relies on animal testing. Embryonic development depends on cell-autonomous and non-autonomous processes including spatiotemporally regulated extracellular signaling activities. These have been elucidated in transgenic mouse models harboring easily detectable reporter genes under the control of evolutionarily conserved signaling cascades. We propose combining these transgenic mouse models and cells derived thereof with existing alternative toxicological testing strategies. This would enable the plausibility of in vitro data to be verified in light of in vivo data and, ultimately, facilitate regulatory acceptance of in vitro test methods. Copyright © 2016 Elsevier Ltd. All rights reserved.

  12. 21 CFR 312.160 - Drugs for investigational use in laboratory research animals or in vitro tests.

    Science.gov (United States)

    2010-04-01

    ... research animals or in vitro tests. 312.160 Section 312.160 Food and Drugs FOOD AND DRUG ADMINISTRATION... Drugs for Investigational Use in Laboratory Research Animals or In Vitro Tests § 312.160 Drugs for investigational use in laboratory research animals or in vitro tests. (a) Authorization to ship. (1)(i) A person...

  13. Evaluation of a new Syphilis assay on Vitros® 5600 Integrated System

    Directory of Open Access Journals (Sweden)

    Giusy Longo

    2010-12-01

    Full Text Available Introduction. A new homogeneous immunoassay for detection of primary infection of Treponema Pallidum (TP on Vitros® 5600 Integrated System was evaluated.The scope of the study was to verify analytical performances and diagnostic accuracy in comparison to commercial methods (Immunoblotting test, ELISA test, Immunoturbidimetric test. Methods. The new Syphilis assay from SENTINEL CH. SpA, is an immunoturbidimetric assay, using microparticles coated with TP fixed on the surface of polystyrene latex particles which agglutinate by an antigen-antibody reaction when anti-TP antigen is present in the specimen. The assay was implemented on Vitros® 5600 Integrated System. Modified CLSI protocols were adopted. Acceptance criteria for total imprecision were 5% for negative samples (or SD 0.5 U/mL and 4% for positive samples. In comparison to commercial methods, sensitivity must be 99.5% and specificity 99.5%. Results. Total imprecision (22 days gave SD at 6 U/mL lower than 0.5 U/mL, and CV% at 10 U/mL and 45 U/mL lower than 4%. Low quantitation limit is 5 U/mL. No prozone up to 13000 U/mL was found. In the on-board calibration stability study no drift was found up to 4 weeks. 153 samples were tested vs immunoblotting method and specificity was 100%, sensitivity was 100%. 495 samples were tested vs ELISA method and test specificity and sensitivity were 99.6% and 100% respectively. 521 samples were tested vs immunoturbidimetric method and specificity was 99.8%, sensitivity was 100%. Interference from Bilirubin (20 mg/dL, Hemoglobin (500 mg/dL and Triglycerides (1000 mg/dL was not detected.All the sample collection tubes tested (K2EDTA, SST, LH PST II, LH, NH did not interfere with the assay. Conclusion. Performances of the new SENTINEL Syphilis assay on Vitros® 5600 Integrated System meet the requirements for its use as screening tool in blood bank, thus allowing consolidation with general chemistry on a single high volume chemistry analyzer, which is

  14. Diagnostic efficacy of in vitro methods vs. skin testing in patients with inhalant allergies

    International Nuclear Information System (INIS)

    Corey, J.P.; Liudahl, J.J.; Young, S.A.; Rodman, S.M.

    1991-01-01

    The purpose of our study was to investigate the diagnostic efficacy of two selected methods of in vitro allergy testing. Specifically, the PRIST/modified RAST I125 isotope systems and the Quantizyme/modified EAST alkaline phosphatase method were compared. The time, expense, convenience, and diagnostic efficacy of the two procedures are discussed. Special attention is given to the practicality of each method for the practicing physician

  15. Validation of Alternative In Vitro Methods to Animal Testing: Concepts, Challenges, Processes and Tools.

    Science.gov (United States)

    Griesinger, Claudius; Desprez, Bertrand; Coecke, Sandra; Casey, Warren; Zuang, Valérie

    This chapter explores the concepts, processes, tools and challenges relating to the validation of alternative methods for toxicity and safety testing. In general terms, validation is the process of assessing the appropriateness and usefulness of a tool for its intended purpose. Validation is routinely used in various contexts in science, technology, the manufacturing and services sectors. It serves to assess the fitness-for-purpose of devices, systems, software up to entire methodologies. In the area of toxicity testing, validation plays an indispensable role: "alternative approaches" are increasingly replacing animal models as predictive tools and it needs to be demonstrated that these novel methods are fit for purpose. Alternative approaches include in vitro test methods, non-testing approaches such as predictive computer models up to entire testing and assessment strategies composed of method suites, data sources and decision-aiding tools. Data generated with alternative approaches are ultimately used for decision-making on public health and the protection of the environment. It is therefore essential that the underlying methods and methodologies are thoroughly characterised, assessed and transparently documented through validation studies involving impartial actors. Importantly, validation serves as a filter to ensure that only test methods able to produce data that help to address legislative requirements (e.g. EU's REACH legislation) are accepted as official testing tools and, owing to the globalisation of markets, recognised on international level (e.g. through inclusion in OECD test guidelines). Since validation creates a credible and transparent evidence base on test methods, it provides a quality stamp, supporting companies developing and marketing alternative methods and creating considerable business opportunities. Validation of alternative methods is conducted through scientific studies assessing two key hypotheses, reliability and relevance of the

  16. In Vitro Exposure Systems and Dosimetry Assessment Tools ...

    Science.gov (United States)

    In 2009, the passing of The Family Smoking Prevention and Tobacco Control Act facilitated the establishment of the FDA Center for Tobacco Products (CTP) and gave it regulatory authority over the marketing, manufacture and distribution of tobacco products, including those termed “modified risk”. On 4-6 April 2016, the Institute for In Vitro Sciences, Inc. (IIVS) convened a workshop conference titled “In Vitro Exposure Systems and Dosimetry Assessment Tools for Inhaled Tobacco Products” to bring together stakeholders representing regulatory agencies, academia, and industry to address the research priorities articulated by the FDA CTP. Specific topics were covered to assess the status of current in vitro smoke and aerosol/vapor exposure systems, as well as the various approaches and challenges to quantifying the complex exposures, in in vitro pulmonary models developed for evaluating adverse pulmonary events resulting from tobacco product exposures. The four core topics covered were, 1) Tobacco Smoke And E-Cigarette Aerosols, 2) Air-Liquid Interface-In Vitro Exposure Systems, 3) Dosimetry Approaches For Particles And Vapors; In Vitro Dosimetry Determinations and 4) Exposure Microenvironment/Physiology Of Cells. The two and a half day workshop included presentations from 20 expert speakers, poster sessions, networking discussions, and breakout sessions which identified key findings and provided recommendations to advance these technologies. Here, we will re

  17. Critical Evaluation of Air-Liquid Interface Cell Exposure Systems for in Vitro Assessment of Atmospheric Pollutants

    Science.gov (United States)

    We compared various in vitro exposure systems for their ability to expose cells to particles and gases. The systems tested use different mechanisms to deliver multi-pollutants to the cells: diffusion, sedimentation, thermophoresis (THP) and electrostatic precipitation (ESP). Vari...

  18. Toxicity assessment of industrial chemicals and airborne contaminants: transition from in vivo to in vitro test methods: a review.

    Science.gov (United States)

    Bakand, S; Winder, C; Khalil, C; Hayes, A

    2005-12-01

    Exposure to occupational and environmental contaminants is a major contributor to human health problems. Inhalation of gases, vapors, aerosols, and mixtures of these can cause a wide range of adverse health effects, ranging from simple irritation to systemic diseases. Despite significant achievements in the risk assessment of chemicals, the toxicological database, particularly for industrial chemicals, remains limited. Considering there are approximately 80,000 chemicals in commerce, and an extremely large number of chemical mixtures, in vivo testing of this large number is unachievable from both economical and practical perspectives. While in vitro methods are capable of rapidly providing toxicity information, regulatory agencies in general are still cautious about the replacement of whole-animal methods with new in vitro techniques. Although studying the toxic effects of inhaled chemicals is a complex subject, recent studies demonstrate that in vitro methods may have significant potential for assessing the toxicity of airborne contaminants. In this review, current toxicity test methods for risk evaluation of industrial chemicals and airborne contaminants are presented. To evaluate the potential applications of in vitro methods for studying respiratory toxicity, more recent models developed for toxicity testing of airborne contaminants are discussed.

  19. In vitro and ex vivo testing of tenofovir shows it is effective as an HIV-1 microbicide.

    Directory of Open Access Journals (Sweden)

    Lisa C Rohan

    2010-02-01

    Full Text Available Tenofovir gel has entered into clinical trials for use as a topical microbicide to prevent HIV-1 infection but has no published data regarding pre-clinical testing using in vitro and ex vivo models. To validate our findings with on-going clinical trial results, we evaluated topical tenofovir gel for safety and efficacy. We also modeled systemic application of tenofovir for efficacy.Formulation assessment of tenofovir gel included osmolality, viscosity, in vitro release, and permeability testing. Safety was evaluated by measuring the effect on the viability of vaginal flora, PBMCs, epithelial cells, and ectocervical and colorectal explant tissues. For efficacy testing, PBMCs were cultured with tenofovir or vehicle control gels and HIV-1 representing subtypes A, B, and C. Additionally, polarized ectocervical and colorectal explant cultures were treated apically with either gel. Tenofovir was added basolaterally to simulate systemic application. All tissues were challenged with HIV-1 applied apically. Infection was assessed by measuring p24 by ELISA on collected supernatants and immunohistochemistry for ectocervical explants. Formulation testing showed the tenofovir and vehicle control gels were >10 times isosmolar. Permeability through ectocervical tissue was variable but in all cases the receptor compartment drug concentration reached levels that inhibit HIV-1 infection in vitro. The gels were non-toxic toward vaginal flora, PBMCs, or epithelial cells. A transient reduction in epithelial monolayer integrity and epithelial fracture for ectocervical and colorectal explants was noted and likely due to the hyperosmolar nature of the formulation. Tenofovir gel prevented HIV-1 infection of PBMCs regardless of HIV-1 subtype. Topical and systemic tenofovir were effective at preventing HIV-1 infection of explant cultures.These studies provide a mechanism for pre-clinical prediction of safety and efficacy of formulated microbicides. Tenofovir was effective

  20. Recommended procedures for performance testing of radiobioassay laboratories: Volume 2, In vitro samples

    International Nuclear Information System (INIS)

    Fenrick, H.W.; MacLellan, J.A.

    1988-11-01

    Draft American National Standards Institute (ANSI) Standard N13.30 (Performance Criteria for Radiobioassay) was developed for the US Department of Energy and the US Nuclear Regulatory Commission to help ensure that bioassay laboratories provide accurate and consistent results. The draft standard specifies the criteria for defining the procedures necessary to establish a bioassay performance-testing laboratory and program. The bioassay testing laboratory will conduct tests to evaluate the performance of service laboratories. Pacific Northwest Laboratory helped develop testing procedures as part of an effort to evaluate the performance criteria by testing the existing measurement capabilities of various bioassay laboratories. This report recommends guidelines for the preparation, handling, storage, distribution, shipping, and documentation of in vitro test samples (artificial urine and fecal matter) for indirect bioassay. The data base and recommended records system for documenting radiobioassay performance at the service laboratories are also presented. 8 refs., 3 tabs

  1. Evaluation of the Ortho-Clinical Diagnostics Vitros ECi Anti-HCV test: comparison with three other methods.

    Science.gov (United States)

    Watterson, Jeannette M; Stallcup, Paulina; Escamilla, David; Chernay, Patrick; Reyes, Alfred; Trevino, Sylvia C

    2007-01-01

    After observing a high incidence of low positive hepatitis C virus (HCV) antibody screens by the Ortho-Clinical Vitros ECi test (Orthoclinical Diagnostics, Raritan, NJ), we compared results against those obtained using another chemiluminescent analyzer, as well as two U.S. Food and Drug Administration (FDA)-approved confirmatory methodologies. To ascertain the true anti-HCV status of samples deemed low-positive by the Ortho-Clinical Vitros ECi test, we tested samples using the ADVIA Centaur HCV screen test (Siemens Medical Solutions Diagnostics), the Chiron recombinant immunoblot assay (RIBA) test (Chiron Corp., Emeryville, CA), and the Roche COBAS Amplicor HCV qualitative test (Roche Diagnostics, Indianapolis, IN) in a series of studies. Of 94 specimens positive by Vitros ECi, 19% were observed to be negative by Centaur. A separate study of 91 samples with signal-to-cutoff (s/co) values less than 8.0 showed that all but one was negative for HCV ribonucleic acid (RNA). In comparison with RIBA, 100% (77) samples positive by the Vitros ECi test with s/co values less than 12.0 were negative or indeterminate by RIBA. A final study comparing all four methods side-by-side showed 63% disagreement by Centaur for Vitros ECi low-positive samples, 75% disagreement by RIBA, and 97% disagreement by polymerase chain reaction (PCR). In conclusion, the Ortho-Clinical Vitros ECi Anti-HCV test yields a high rate of false-positive results in the low s/co range in our patient population. (c) 2007 Wiley-Liss, Inc.

  2. A comparison of finite element analysis with in vitro bond strength tests of the bracket-cement-enamel system

    NARCIS (Netherlands)

    Algera, T.J.; Feilzer, A.J.; Prahl-Andersen, B.; Kleverlaan, C.J.

    2011-01-01

    The aim of this study was to determine the in vitro shear bond strength (SBS) and tensile bond strength (TBS) of 45 metal brackets bonded with Transbond XT to bovine enamel. The SBS was determined by loading the short and the long sides of the bracket base. Testing took place after storage of the

  3. In vitro binding of estrogens by dietary fiber and the in vivo apparent digestibility tested in pigs

    NARCIS (Netherlands)

    Arts, C.J.M.; Govers, C.A.R.L.; Berg, H. van den; Wolters, M.G.E.; Leeuwen, P. van; Thijsen, J.H.

    1991-01-01

    Within the framework of experiments related to the association between dietary fiber and breast cancer an in vitro test system was used to study the binding of estrogens to various fibers (e.g. cholestyramin, lignin and cellulose) and fiber sources (e.g. wheat bran, cereals, seeds and legumes).

  4. In vitro Alternative Methodologies for Central Nervous System Assessment: A Critique using Nanoscale Materials as an Example.

    Science.gov (United States)

    Identifying the potential health hazards to the central nervous system of a new family of materials presents many challenges. Whole-animal toxicity testing has been the tradition, but in vitro methods have been steadily gaining popularity. There are numerous challenges in testing...

  5. A reproducible accelerated in vitro release testing method for PLGA microspheres.

    Science.gov (United States)

    Shen, Jie; Lee, Kyulim; Choi, Stephanie; Qu, Wen; Wang, Yan; Burgess, Diane J

    2016-02-10

    The objective of the present study was to develop a discriminatory and reproducible accelerated in vitro release method for long-acting PLGA microspheres with inner structure/porosity differences. Risperidone was chosen as a model drug. Qualitatively and quantitatively equivalent PLGA microspheres with different inner structure/porosity were obtained using different manufacturing processes. Physicochemical properties as well as degradation profiles of the prepared microspheres were investigated. Furthermore, in vitro release testing of the prepared risperidone microspheres was performed using the most common in vitro release methods (i.e., sample-and-separate and flow through) for this type of product. The obtained compositionally equivalent risperidone microspheres had similar drug loading but different inner structure/porosity. When microsphere particle size appeared similar, porous risperidone microspheres showed faster microsphere degradation and drug release compared with less porous microspheres. Both in vitro release methods investigated were able to differentiate risperidone microsphere formulations with differences in porosity under real-time (37 °C) and accelerated (45 °C) testing conditions. Notably, only the accelerated USP apparatus 4 method showed good reproducibility for highly porous risperidone microspheres. These results indicated that the accelerated USP apparatus 4 method is an appropriate fast quality control tool for long-acting PLGA microspheres (even with porous structures). Copyright © 2015 Elsevier B.V. All rights reserved.

  6. Chapter 31: Common in vitro tests for allergy and immunology.

    Science.gov (United States)

    Makhija, Melanie; O'Gorman, Maurice R G

    2012-01-01

    Allergen-specific IgE antibody is the most commonly ordered in vitro test in the practice of allergy and is used to diagnose type I hypersensitivity reactions to foods or reactivity to aeroallergens in patients with relative contraindications to skin-prick testing such as dermatographism. The Phadebas radioallergosorbent test (RAST; Pharmacia, Uppsala, Sweden) was the first assay reported for the detection of the allergen-specific IgE antibody. In a RAST, antigen (allergen) is bound to a solid phase, such as a paper disk, and then incubated with human serum. A buffer wash removes unbound serum proteins, and radiolabeled anti-human IgE is added to detect bound IgE, if present. The results are reported in arbitrary units of IgE per milliliter of serum. The term RAST was originally a brand name but it is now often used colloquially (and incorrectly) to describe any in vitro assay for allergen-specific IgE. Total serum IgE can be measured and is helpful in determining atopic presentations such as in allergic bronchopulmonary aspergillosis or in patients with persistent asthma who are candidates for monoclonal anti-IgE antibody therapy with, omalizumab. In patients with recurrent bacterial infections of the sinopulmonary tract, the basic humoral immune system testing includes measuring quantitative immunoglobulins (IgG, IgA, and IgM) and comparing them to age-matched normal ranges. Most clinical laboratories use nephelometry to measure immunoglobulin levels quantitatively. Nephelometry detects either the rate or the end point of soluble immune complex formation (the IgG in sera complexes with an anti-IgG antibody forming a classic immunoprecipitation reaction) by monitoring the scatter of transmitted light. The most common method for the screening of cellular immunodeficiency involved the measurement of the absolute and relative representation of the major lymphocyte subsets, T-cells, T-helper cells, T-cytotoxic cells, B-cells and NK-cells.

  7. The sensitivity testing of Wilms' tumors to cytostatic agents with an autoradiographic in vitro short-term test

    International Nuclear Information System (INIS)

    Willnow, U.

    1984-01-01

    Sensitivity of 15 Wilms' tumors in children was tested towards cytostatic agents in vitro by means of an autoradiographic short-term test. Sensitivity was measured as the magnitude of the inhibition of 3 H-thymidine or 3 H-uridine incorporation. The test was performed with Adriamycin, Actinomycin D, Daunomycin, Bleomycin, Cyclophosphamide, Ifosfamide, Trenimon, and Arabinosylcytosine. None of the tumors is resistant to all substances, they are responsive against 2 or more drugs. The most effective drugs tested are Adriamycin, Actinomycin D and Cyclophosphamide. The tumors show a marked individual sensitivity pattern. This behavior is explained mainly by the usually high proliferative activity of Wilms' tumors. The possibilities and limits of long-term and short-term methods for sensitivity testing are discussed critically. For the evaluation of the results of in vitro testing and in vivo effectiveness the close correlation should be considered between the type of cytostatic agent and proliferation kinetics of the tumor, cytostatic agent and effect on tumor metabolism as well as the effect of the cytostatics and the nucleic acid precursors used for the short-term test. Despite the methodological limitations preclinical testing should be preferred to unselected chemotherapy. (author)

  8. In vitro antitumour activity, safety testing and subcellular distribution of two poly[oxyethylene(aminophosphonate-co-H-phosphonate]s in Ehrlich ascites carcinoma and BALB/c 3T3 cell culture systems

    Directory of Open Access Journals (Sweden)

    Ani Georgieva

    2016-01-01

    Full Text Available Two polyphosphoesters containing anthracene-derived aminophosphonate and hydrophilic H-phosphonate repeating units, poly[oxyethylene(aminophosphonate-co-H-phosphonate]s (1 and 2, were tested for the in vitro antitumour activity on cell cultures derived from ascitic form of Ehrlich mammary adenocarcinoma by 3-(4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (MTT-dye reduction assay. The in vitro safety testing of the copolymers was performed by BALB/c 3T3 neutral red uptake assay. A study on their uptake and subcellular distribution in non-tumourigenic and tumour cells was performed by means of fluorescence microscopy. Both copolymers showed significant antitumour activity towards Ehrlich ascites carcinoma (EAC cells. However, the in vitro safety testing revealed significant toxicity of polymer 2 to BALB/c 3T3 mouse embryo cells. In contrast, polymer 1 showed complete absence of cytotoxicity to BALB/c 3T3 cells. The fluorescent studies showed that the substances were diffusely distributed in the cytoplasm in both cell culture systems. As opposed to BALB/c 3T3 cells, in EAC cells, intense fluorescent signal was observed in the nuclei and in the perinuclear region. The tested polyphosphoesters are expected to act under physiological conditions as prodrugs of aminophosphonates.

  9. In vitro chemical and cellular tests applied to uranium trioxide with different hydration states

    International Nuclear Information System (INIS)

    Ansoborlo, E.; Chalabreysse, J.; Henge-Napoli, M.H.; Pujol, E.

    1992-01-01

    A simple and rapid in vitro chemical solubility test applicable to industrial uranium trioxide (UO 3 ) was developed together with two in vitro cellular tests using rat alveolar macrophages maintained either in gas phase or in alginate beads at 37 degrees C. Industrial UO 3 was characterized by particle size, X-ray, and IR spectra, and chemical transformation (e.g., aging and hydration of the dust) was also studied. Solvents used for the in vitro chemical solubility study included carbonates, citrates, phosphates, water, Eagle's basal medium, and Gamble's solution (simulated lung fluid), alone, with oxygen, or with superoxide ions. Results, expressed in terms of the half-time of dissolution, according to International Commission on Radiological Protection (ICRP) classification (D,W,Y), varied for different hydration states of UO 3 , showing a lower solubility of hydrated UO 3 in solvents compared to basic UO 3 or UO 3 heated at 450 degrees C. Two in vitro cellular tests on cultured rat alveolar macrophages (cells maintained in gas phase and cells immobilized in alginate beads) were used on the same UO 3 samples and generally showed a lower solution transfer rate in the presence of macrophages than in the culture medium alone. The results of in vitro chemical and cellular tests were compared, with four main conclusions; a good reproducibility of the three tests in Eagle's basal medium of the effect of hydration state on solubility, the classification of UO 3 in terms of ICRP solubility criteria, and the ability of macrophoges to decrease uranium solubility in medium. 16 refs., 3 figs., 4 tabs

  10. In vitro toxicities of experimental jet fuel system ice-inhibiting agents.

    Science.gov (United States)

    Geiss, K T; Frazier, J M

    2001-07-02

    One research emphasis within the Department of Defense has been to seek the replacement of operational compounds with alternatives that pose less potential risk to human and ecological systems. Alternatives to glycol ethers, such as diethylene glycol monomethyl ether (M-DE), were investigated for use as jet fuel system ice-inhibiting agents (FSIIs). This group of chemicals includes three derivatives of 1,3-dioxolane-4-methanol (M-1, M-2, and M-3) and a 1,3-dioxane (M-27). In addition, M-DE was evaluated as a reference compound. Our approach was to implement an in vitro test battery based on primary rat hepatocyte cultures to perform initial toxicity evaluations. Hepatocytes were exposed to experimental chemicals (0, 0.001, 0.01, 0.1, 1, 10 mM dosages) for periods up to 24 h. Samples were assayed for lactate dehydrogenase (LDH) release, MTT dye reduction activity, glutathione level, and rate of protein synthesis as indicators of toxicity. Of the compounds tested, M-1, especially at the 10-mM dose, appeared to be more potent than the other chemicals, as measured by these toxicity assays. M-DE, the current FSII, elicited little response in the toxicity assays. Although some variations in toxicity were observed at the 10-mM dose, the in vitro toxicities of the chemicals tested (except for M-1) were not considerably greater than that of M-DE.

  11. The use of metabolising systems for in vitro testing of endocrine disruptors

    NARCIS (Netherlands)

    Jacobs, M.N.; Janssens, W.; Bernauer, U.; Brandon, E.; Coecke, S.; Combes, R.; Edwards, P.; Freidig, A.; Freyberger, A.; Kolanczyk, R.; Mc Ardie, C.; Mekenyan, O.; Schmieder, P.; Schrader, T.; Takeyoshi, M.; Burg, B. van der

    2008-01-01

    Legislation and prospective legislative proposals in for instance the USA, Europe, and Japan require, or may require that chemicals are tested for their ability to disrupt the hormonal systems of mammals. Chemicals found to test positive are considered to be endocrine active substances (EAS) and may

  12. Test systems in drug discovery for hazard identification and risk assessment of human drug-induced liver injury.

    Science.gov (United States)

    Weaver, Richard J; Betts, Catherine; Blomme, Eric A G; Gerets, Helga H J; Gjervig Jensen, Klaus; Hewitt, Philip G; Juhila, Satu; Labbe, Gilles; Liguori, Michael J; Mesens, Natalie; Ogese, Monday O; Persson, Mikael; Snoeys, Jan; Stevens, James L; Walker, Tracy; Park, B Kevin

    2017-07-01

    The liver is an important target for drug-induced toxicities. Early detection of hepatotoxic drugs requires use of well-characterized test systems, yet current knowledge, gaps and limitations of tests employed remains an important issue for drug development. Areas Covered: The current state of the science, understanding and application of test systems in use for the detection of drug-induced cytotoxicity, mitochondrial toxicity, cholestasis and inflammation is summarized. The test systems highlighted herein cover mostly in vitro and some in vivo models and endpoint measurements used in the assessment of small molecule toxic liabilities. Opportunities for research efforts in areas necessitating the development of specific tests and improved mechanistic understanding are highlighted. Expert Opinion: Use of in vitro test systems for safety optimization will remain a core activity in drug discovery. Substantial inroads have been made with a number of assays established for human Drug-induced Liver Injury. There nevertheless remain significant gaps with a need for improved in vitro tools and novel tests to address specific mechanisms of human Drug-Induced Liver Injury. Progress in these areas will necessitate not only models fit for application, but also mechanistic understanding of how chemical insult on the liver occurs in order to identify translational and quantifiable readouts for decision-making.

  13. Assessment of cosmetic ingredients in the in vitro reconstructed human epidermis test method EpiSkin™ using HPLC/UPLC-spectrophotometry in the MTT-reduction assay.

    Science.gov (United States)

    Alépée, N; Hibatallah, J; Klaric, M; Mewes, K R; Pfannenbecker, U; McNamee, P

    2016-06-01

    Cosmetics Europe recently established HPLC/UPLC-spectrophotometry as a suitable alternative endpoint detection system for measurement of formazan in the MTT-reduction assay of reconstructed human tissue test methods irrespective of the test system involved. This addressed a known limitation for such test methods that use optical density for measurement of formazan and may be incompatible for evaluation of strong MTT reducer and/or coloured chemicals. To build on the original project, Cosmetics Europe has undertaken a second study that focuses on evaluation of chemicals with functionalities relevant to cosmetic products. Such chemicals were primarily identified from the Scientific Committee on Consumer Safety (SCCS) 2010 memorandum (addendum) on the in vitro test EpiSkin™ for skin irritation testing. Fifty test items were evaluated in which both standard photometry and HPLC/UPLC-spectrophotometry were used for endpoint detection. The results obtained in this study: 1) provide further support for Within Laboratory Reproducibility of HPLC-UPLC-spectrophotometry for measurement of formazan; 2) demonstrate, through use a case study with Basazol C Blue pr. 8056, that HPLC/UPLC-spectrophotometry enables determination of an in vitro classification even when this is not possible using standard photometry and 3) addresses the question raised by SCCS in their 2010 memorandum (addendum) to consider an endpoint detection system not involving optical density quantification in in vitro reconstructed human epidermis skin irritation test methods. Copyright © 2016 Elsevier Ltd. All rights reserved.

  14. A ring test of a wireless in vitro gas production system

    DEFF Research Database (Denmark)

    Cornou, Cecile; Storm, Ida Marie Lindhardt Drejer; Hindrichsen, Ida Katarina Auf der M.

    2013-01-01

    ) or sheep (SP). Curves, corrected for blanks, were fitted using an exponential regression model with a lag time. The following variables were considered: (i) GP24 and GP48: raw values at 24 and 48 h (mL/g DM), corrected for blanks; (ii) A: asymptotic GP (mL/g DM); (iii) T1/2: time when half A is produced (h......); (iv) GPMR: maximum predicted GP rate (mL/h); (v) L: lag time (h). A mixed model including laboratories as random effect was used. A significant interaction between substrate and laboratories was found for all variables except A. The most repeatable and reproducible results were observed for A and GP48......The in vitro gas production (GP) technique has been widely used for feed evaluation. However, variability in results limits useful comparisons. Results from a ring test undertaken in four laboratories (Italy - IT, Spain - SP, Wales - WA and Denmark - DK) using the same wireless equipment (ANKOM...

  15. Anticoccidial efficacy testing: In vitro Eimeria tenella assays as replacement for animal experiments.

    Science.gov (United States)

    Thabet, Ahmed; Zhang, Runhui; Alnassan, Alaa-Aldin; Daugschies, Arwid; Bangoura, Berit

    2017-01-15

    Availability of an accurate in vitro assay is a crucial demand to determine sensitivity of Eimeria spp. field strains toward anticoccidials routinely. In this study we tested in vitro models of Eimeria tenella using various polyether ionophores (monensin, salinomycin, maduramicin, and lasalocid) and toltrazuril. Minimum inhibitory concentrations (MIC 95 , MIC 50/95 ) for the tested anticoccidials were defined based on a susceptible reference (Houghton strain), Ref-1. In vitro sporozoite invasion inhibition assay (SIA) and reproduction inhibition assay (RIA) were applied on sensitive laboratory (Ref-1 and Ref-2) and field (FS-1, FS-2, and FS-3) strains to calculate percent of inhibition under exposure of these strains to the various anticoccidials (%I SIA and%I RIA, respectively). The in vitro data were related to oocyst excretion, lesion scores, performance, and global resistance indices (GI) assessed in experimentally infected chickens. Polyether ionophores applied in the RIA were highly effective at MIC 95 against Ref-1 and Ref-2 (%I RIA ≥95%). In contrast, all tested field strains displayed reduced to low efficacy (%I RIA animal model (p89%) against all strains used in this study. However, adjusted GI (GI adj ) for toltrazuril-treated groups exhibited differences between reference and field strains which might indicate varying sensitivity. RIA is a suitable in vitro tool to detect sensitivity of E. tenella towards polyether ionophores, and may thus help to reduce, replace, or refine use of animal experimentation for in vivo sensitivity assays. Copyright © 2016 Elsevier B.V. All rights reserved.

  16. In vitro preliminary cytotoxicity testing of vegetal extracts, using colorimetric methods

    Directory of Open Access Journals (Sweden)

    Claudia Patricia Cordero Camacho

    2002-01-01

    Full Text Available To advance in the study of the Colombian vegetal biodiversity, considered as a potential source of pharmacologically active products, the establishment of biological activity evaluation systems is necessary, which allow the detection of active products against pathologies with high social and economical impact, such as cancer. This work describes the implementation of a preliminary in vitro methodology for the determination of potential anticancer activity in vegetal extracts, by cytotoxicity testing upon human tumor cell lines, measuring the cellular mass indirectly with the colorimetric assays of MTT (methyl tetrazolium tiazole reduction and SRB (sulforhodamine Bstaining. HT-29, MCF-7, SiHa and HEp-2 cell lines cultures were adapted, MTT concentration, cellular density and treatment period parameters for the cytotoxicity assay were selected. Cell lines sensitivity to the chemotherapeutic agent Doxorubicin HCl was determined. Colombian vegetal species extracts cytotoxicity was tested and usefulness of the assay as a tool to bioguide the search of active products was evidenced.

  17. In vitro preliminary cytotoxicity testing of vegetal extracts, using colorimetric methods

    Directory of Open Access Journals (Sweden)

    Claudia Patricia Cordero Camacho

    2011-12-01

    Full Text Available To advance in the study of the Colombian vegetal biodiversity, considered as a potential source of pharmacologically active products, the establishment of biological activity evaluation systems is necessary, which allow the detection of active products against pathologies with high social and economical impact, such as cancer. This work describes the implementation of a preliminary in vitro methodology for the determination of potential anticancer activity in vegetal extracts, by cytotoxicity testing upon human tumor cell lines, measuring the cellular mass indirectly with the colorimetric assays of MTT (methyl tetrazolium tiazole reduction and SRB (sulforhodamine Bstaining. HT-29, MCF-7, SiHa and HEp-2 cell lines cultures were adapted, MTT concentration, cellular density and treatment period parameters for the cytotoxicity assay were selected. Cell lines sensitivity to the chemotherapeutic agent Doxorubicin HCl was determined. Colombian vegetal species extracts cytotoxicity was tested and usefulness of the assay as a tool to bioguide the search of active products was evidenced.

  18. In-vitro orthodontic bond strength testing : A systematic review and meta-analysis

    NARCIS (Netherlands)

    Finnema, K.J.; Ozcan, M.; Post, W.J.; Ren, Y.J.; Dijkstra, P.U.

    INTRODUCTION: The aims of this study were to systematically review the available literature regarding in-vitro orthodontic shear bond strength testing and to analyze the influence of test conditions on bond strength. METHODS: Our data sources were Embase and Medline. Relevant studies were selected

  19. A strategy for in vitro safety testing of nanotitania-modified textile products

    International Nuclear Information System (INIS)

    Roszak, Joanna; Stępnik, Maciej; Nocuń, Marek; Ferlińska, Magdalena; Smok-Pieniążek, Anna; Grobelny, Jarosław; Tomaszewska, Emilia; Wąsowicz, Wojciech; Cieślak, Małgorzata

    2013-01-01

    Highlights: • Commercially available TiO 2 /Ag nanomaterials (NMs) showed higher cytotoxic effect than TiO 2 NMs. • Both titania NMs in pristine form induced a weak genotoxic effect in in vitro studies. • Cytotoxic effect of textile materials modified with TiO 2 /Ag NMs depended on the mode of the fiber manufacturing. • The strategy of in vitro testing of textile materials modified with NMs was proposed. -- Abstract: Titanium dioxide nanomaterials are extensively used in many applications, also for modification of textile materials. Toxicological assessment of such textile materials is currently seldom performed, mainly because of lack of appropriate guidelines. The aim of the study was to assess cytotoxic and genotoxic potential of commercially available TiO 2 and TiO 2 /Ag NMs in pristine form as well as polypropylene fibers modified with the NMs. Both titania NMs showed a cytotoxic effect on BALB/3T3 clone A31 and V79 fibroblasts after 72-h exposure. Both NMs induced a weak genotoxic effect in comet assay, with TiO 2 /Ag being more active. In vitro micronucleus test on human lymphocytes revealed a weak mutagenic effect of both materials after 24 h of exposure. In contrast, no significant increase in micronuclei frequency was observed in the in vitro micronucleus test on V79 fibroblasts. The 24-h extracts prepared from polypropylene fibers modified with TiO 2 /Ag induced a cytotoxic effect on BALB/3T3 cells which strongly depended on the mode of the fibers manufacturing. The study presents a comprehensive approach to toxicity assessment of textile fibers modified with NMs. Proposed approach may form a good “starting point” for improved future testing strategies

  20. A strategy for in vitro safety testing of nanotitania-modified textile products

    Energy Technology Data Exchange (ETDEWEB)

    Roszak, Joanna; Stępnik, Maciej; Nocuń, Marek; Ferlińska, Magdalena; Smok-Pieniążek, Anna [Nofer Institute of Occupational Medicine, 8 St Teresy St., 91-348 Łódź (Poland); Grobelny, Jarosław; Tomaszewska, Emilia [University of Lodz, Faculty of Chemistry, 163 Pomorska St, 90-236 Łódź (Poland); Wąsowicz, Wojciech [Nofer Institute of Occupational Medicine, 8 St Teresy St., 91-348 Łódź (Poland); Cieślak, Małgorzata, E-mail: cieslakm@iw.lodz.pl [Textile Research Institute, 118 Gdańska St., 90-520, Łódź (Poland)

    2013-07-15

    Highlights: • Commercially available TiO{sub 2}/Ag nanomaterials (NMs) showed higher cytotoxic effect than TiO{sub 2} NMs. • Both titania NMs in pristine form induced a weak genotoxic effect in in vitro studies. • Cytotoxic effect of textile materials modified with TiO{sub 2}/Ag NMs depended on the mode of the fiber manufacturing. • The strategy of in vitro testing of textile materials modified with NMs was proposed. -- Abstract: Titanium dioxide nanomaterials are extensively used in many applications, also for modification of textile materials. Toxicological assessment of such textile materials is currently seldom performed, mainly because of lack of appropriate guidelines. The aim of the study was to assess cytotoxic and genotoxic potential of commercially available TiO{sub 2} and TiO{sub 2}/Ag NMs in pristine form as well as polypropylene fibers modified with the NMs. Both titania NMs showed a cytotoxic effect on BALB/3T3 clone A31 and V79 fibroblasts after 72-h exposure. Both NMs induced a weak genotoxic effect in comet assay, with TiO{sub 2}/Ag being more active. In vitro micronucleus test on human lymphocytes revealed a weak mutagenic effect of both materials after 24 h of exposure. In contrast, no significant increase in micronuclei frequency was observed in the in vitro micronucleus test on V79 fibroblasts. The 24-h extracts prepared from polypropylene fibers modified with TiO{sub 2}/Ag induced a cytotoxic effect on BALB/3T3 cells which strongly depended on the mode of the fibers manufacturing. The study presents a comprehensive approach to toxicity assessment of textile fibers modified with NMs. Proposed approach may form a good “starting point” for improved future testing strategies.

  1. Assuring safety without animal testing: the case for the human testis in vitro.

    Science.gov (United States)

    Chapin, Robert E; Boekelheide, Kim; Cortvrindt, Rita; van Duursen, Majorie B M; Gant, Tim; Jegou, Bernard; Marczylo, Emma; van Pelt, Ans M M; Post, Janine N; Roelofs, Maarke J E; Schlatt, Stefan; Teerds, Katja J; Toppari, Jorma; Piersma, Aldert H

    2013-08-01

    From 15 to 17 June 2011, a dedicated workshop was held on the subject of in vitro models for mammalian spermatogenesis and their applications in toxicological hazard and risk assessment. The workshop was sponsored by the Dutch ASAT initiative (Assuring Safety without Animal Testing), which aims at promoting innovative approaches toward toxicological hazard and risk assessment on the basis of human and in vitro data, and replacement of animal studies. Participants addressed the state of the art regarding human and animal evidence for compound mediated testicular toxicity, reviewed existing alternative assay models, and brainstormed about future approaches, specifically considering tissue engineering. The workshop recognized the specific complexity of testicular function exemplified by dedicated cell types with distinct functionalities, as well as different cell compartments in terms of microenvironment and extracellular matrix components. This complexity hampers quick results in the realm of alternative models. Nevertheless, progress has been achieved in recent years, and innovative approaches in tissue engineering may open new avenues for mimicking testicular function in vitro. Although feasible, significant investment is deemed essential to be able to bring new ideas into practice in the laboratory. For the advancement of in vitro testicular toxicity testing, one of the most sensitive end points in regulatory reproductive toxicity testing, such an investment is highly desirable. Copyright © 2013. Published by Elsevier Inc. All rights reserved.

  2. In vitro validation of a novel mechanical model for testing the anchorage capacity of pedicle screws using physiological load application.

    Science.gov (United States)

    Liebsch, Christian; Zimmermann, Julia; Graf, Nicolas; Schilling, Christoph; Wilke, Hans-Joachim; Kienle, Annette

    2018-01-01

    Biomechanical in vitro tests analysing screw loosening often include high standard deviations caused by high variabilities in bone mineral density and pedicle geometry, whereas standardized mechanical models made of PU foam often do not integrate anatomical or physiological boundary conditions. The purpose of this study was to develop a most realistic mechanical model for the standardized and reproducible testing of pedicle screws regarding the resistance against screw loosening and the holding force as well as to validate this model by in vitro experiments. The novel mechanical testing model represents all anatomical structures of a human vertebra and is consisting of PU foam to simulate cancellous bone, as well as a novel pedicle model made of short carbon fibre filled epoxy. Six monoaxial cannulated pedicle screws (Ø6.5 × 45mm) were tested using the mechanical testing model as well as human vertebra specimens by applying complex physiological cyclic loading (shear, tension, and bending; 5Hz testing frequency; sinusoidal pulsating forces) in a dynamic materials testing machine with stepwise increasing load after each 50.000 cycles (100.0N shear force + 20.0N per step, 51.0N tension force + 10.2N per step, 4.2Nm bending moment + 0.8Nm per step) until screw loosening was detected. The pedicle screw head was fixed on a firmly clamped rod while the load was applied in the vertebral body. For the in vitro experiments, six human lumbar vertebrae (L1-3, BMD 75.4 ± 4.0mg/cc HA, pedicle width 9.8 ± 0.6mm) were tested after implanting pedicle screws under X-ray control. Relative motions of pedicle screw, specimen fixture, and rod fixture were detected using an optical motion tracking system. Translational motions of the mechanical testing model experiments in the point of load introduction (0.9-2.2mm at 240N shear force) were reproducible within the variation range of the in vitro experiments (0.6-3.5mm at 240N shear force). Screw loosening occurred continuously in

  3. In vitro testing of thiolated poly(aspartic acid) from ophthalmic formulation aspects.

    Science.gov (United States)

    Budai-Szű Cs, Mária; Horvát, Gabriella; Gyarmati, Benjámin; Szilágyi, Barnabás Áron; Szilágyi, András; Csihi, Tímea; Berkó, Szilvia; Szabó-Révész, Piroska; Mori, Michela; Sandri, Giuseppina; Bonferoni, Maria Cristina; Caramella, Carla; Csányi, Erzsébet

    2016-08-01

    Ocular drug delivery formulations must meet anatomical, biopharmaceutical, patient-driven and regulatory requirements. Mucoadhesive polymers can serve as a better alternative to currently available ophthalmic formulations by providing improved bioavailability. If all requirements are addressed, a polymeric formulation resembling the tear film of the eye might be the best solution. The optimum formulation must not have high osmotic activity, should provide appropriate surface tension, pH and refractive index, must be non-toxic and should be transparent and mucoadhesive. We would like to highlight the importance of in vitro polymer testing from a pharmaceutical aspect. We, therefore, carried out physical-chemical investigations to verify the suitability of certain systems for ophthalmic formulations. In this work, in situ gelling, mucoadhesive thiolated poly(aspartic acid)s were tested from ophthalmic formulation aspects. The results of preformulation measurements indicate that these polymers can be used as potential carriers in ophthalmic drug delivery.

  4. Use of HPLC/UPLC-spectrophotometry for detection of formazan in in vitro Reconstructed human Tissue (RhT)-based test methods employing the MTT-reduction assay to expand their applicability to strongly coloured test chemicals.

    Science.gov (United States)

    Alépée, N; Barroso, J; De Smedt, A; De Wever, B; Hibatallah, J; Klaric, M; Mewes, K R; Millet, M; Pfannenbecker, U; Tailhardat, M; Templier, M; McNamee, P

    2015-06-01

    A number of in vitro test methods using Reconstructed human Tissues (RhT) are regulatory accepted for evaluation of skin corrosion/irritation. In such methods, test chemical corrosion/irritation potential is determined by measuring tissue viability using the photometric MTT-reduction assay. A known limitation of this assay is possible interference of strongly coloured test chemicals with measurement of formazan by absorbance (OD). To address this, Cosmetics Europe evaluated use of HPLC/UPLC-spectrophotometry as an alternative formazan measurement system. Using the approach recommended by the FDA guidance for validation of bio-analytical methods, three independent laboratories established and qualified their HPLC/UPLC-spectrophotometry systems to reproducibly measure formazan from tissue extracts. Up to 26 chemicals were then tested in RhT test systems for eye/skin irritation and skin corrosion. Results support that: (1) HPLC/UPLC-spectrophotometry formazan measurement is highly reproducible; (2) formazan measurement by HPLC/UPLC-spectrophotometry and OD gave almost identical tissue viabilities for test chemicals not exhibiting colour interference nor direct MTT reduction; (3) independent of the test system used, HPLC/UPLC-spectrophotometry can measure formazan for strongly coloured test chemicals when this is not possible by absorbance only. It is therefore recommended that HPLC/UPLC-spectrophotometry to measure formazan be included in the procedures of in vitro RhT-based test methods, irrespective of the test system used and the toxicity endpoint evaluated to extend the applicability of these test methods to strongly coloured chemicals. Copyright © 2015 Elsevier Ltd. All rights reserved.

  5. In vitro to in vivo extrapolation of effective dosimetry in developmental toxicity testing : Application of a generic PBK modelling approach

    NARCIS (Netherlands)

    Fragki, Styliani; Piersma, Aldert H; Rorije, Emiel; Zeilmaker, Marco J

    2017-01-01

    Incorporation of kinetics to quantitative in vitro to in vivo extrapolations (QIVIVE) is a key step for the realization of a non-animal testing paradigm, in the sphere of regulatory toxicology. The use of Physiologically-Based Kinetic (PBK) modelling for determining systemic doses of chemicals at

  6. In vitro to in vivo extrapolation of effective dosimetry in developmental toxicity testing: Application of a generic PBK modelling approach.

    NARCIS (Netherlands)

    Fragki, Styliani; Piersma, Aldert H; Rorije, Emiel; Zeilmaker, Marco J

    2017-01-01

    Incorporation of kinetics to quantitative in vitro to in vivo extrapolations (QIVIVE) is a key step for the realization of a non-animal testing paradigm, in the sphere of regulatory toxicology. The use of Physiologically-Based Kinetic (PBK) modelling for determining systemic doses of chemicals at

  7. In vitro irradiation system for radiobiological experiments

    International Nuclear Information System (INIS)

    Tesei, Anna; Zoli, Wainer; D’Errico, Vincenzo; Romeo, Antonino; Parisi, Elisabetta; Polico, Rolando; Sarnelli, Anna; Arienti, Chiara; Menghi, Enrico; Medri, Laura; Gabucci, Elisa; Pignatta, Sara; Falconi, Mirella; Silvestrini, Rosella

    2013-01-01

    Although two-dimensional (2-D) monolayer cell cultures provide important information on basic tumor biology and radiobiology, they are not representative of the complexity of three-dimensional (3-D) solid tumors. In particular, new models reproducing clinical conditions as closely as possible are needed for radiobiological studies to provide information that can be translated from bench to bedside. We developed a novel system for the irradiation, under sterile conditions, of 3-D tumor spheroids, the in vitro model considered as a bridge between the complex architectural organization of in vivo tumors and the very simple one of in vitro monolayer cell cultures. The system exploits the same equipment as that used for patient treatments, without the need for dedicated and highly expensive instruments. To mimic the passage of radiation beams through human tissues before they reach the target tumor mass, 96-multiwell plates containing the multicellular tumor spheroids (MCTS) are inserted into a custom-built phantom made of plexiglass, the material most similar to water, the main component of human tissue. The system was used to irradiate CAEP- and A549-derived MCTS, pre-treated or not with 20 μM cisplatin, with a dose of 20 Gy delivered in one session. We also tested the same treatment schemes on monolayer CAEP and A549 cells. Our preliminary results indicated a significant increment in radiotoxicity 20 days after the end of irradiation in the CAEP spheroids pre-treated with cisplatin compared to those treated with cisplatin or irradiation alone. Conversely, the effect of the radio- chemotherapy combination in A549-derived MCTS was similar to that induced by cisplatin or irradiation alone. Finally, the 20 Gy dose did not affect cell survival in monolayer CAEP and A549 cells, whereas cisplatin or cisplatin plus radiation caused 100% cell death, regardless of the type of cell line used. We set up a system for the irradiation, under sterile conditions, of tumor cells

  8. Rapid prototyping compliant arterial phantoms for in-vitro studies and device testing

    Directory of Open Access Journals (Sweden)

    Biglino Giovanni

    2013-01-01

    Full Text Available Abstract Background Compliant vascular phantoms are desirable for in-vitro patient-specific experiments and device testing. TangoPlus FullCure 930® is a commercially available rubber-like material that can be used for PolyJet rapid prototyping. This work aims to gather preliminary data on the distensibility of this material, in order to assess the feasibility of its use in the context of experimental cardiovascular modelling. Methods The descending aorta anatomy of a volunteer was modelled in 3D from cardiovascular magnetic resonance (CMR images and rapid prototyped using TangoPlus. The model was printed with a range of increasing wall thicknesses (0.6, 0.7, 0.8, 1.0 and 1.5 mm, keeping the lumen of the vessel constant. Models were also printed in both vertical and horizontal orientations, thus resulting in a total of ten specimens. Compliance tests were performed by monitoring pressure variations while gradually increasing and decreasing internal volume. Knowledge of distensibility was thus derived and then implemented with CMR data to test two applications. Firstly, a patient-specific compliant model of hypoplastic aorta suitable for connection in a mock circulatory loop for in-vitro tests was manufactured. Secondly, the right ventricular outflow tract (RVOT of a patient necessitating pulmonary valve replacement was printed in order to physically test device insertion and assess patient’s suitability for percutaneous pulmonary valve intervention. Results The distensibility of the material was identified in a range from 6.5 × 10-3 mmHg-1 for the 0.6 mm case, to 3.0 × 10-3 mmHg-1 for the 1.5 mm case. The models printed in the vertical orientation were always more compliant than their horizontal counterpart. Rapid prototyping of a compliant hypoplastic aorta and of a RVOT anatomical model were both feasible. Device insertion in the RVOT model was successful. Conclusion Values of distensibility, compared with literature data, show that Tango

  9. Rapid prototyping compliant arterial phantoms for in-vitro studies and device testing.

    Science.gov (United States)

    Biglino, Giovanni; Verschueren, Peter; Zegels, Raf; Taylor, Andrew M; Schievano, Silvia

    2013-01-16

    Compliant vascular phantoms are desirable for in-vitro patient-specific experiments and device testing. TangoPlus FullCure 930 is a commercially available rubber-like material that can be used for PolyJet rapid prototyping. This work aims to gather preliminary data on the distensibility of this material, in order to assess the feasibility of its use in the context of experimental cardiovascular modelling. The descending aorta anatomy of a volunteer was modelled in 3D from cardiovascular magnetic resonance (CMR) images and rapid prototyped using TangoPlus. The model was printed with a range of increasing wall thicknesses (0.6, 0.7, 0.8, 1.0 and 1.5 mm), keeping the lumen of the vessel constant. Models were also printed in both vertical and horizontal orientations, thus resulting in a total of ten specimens. Compliance tests were performed by monitoring pressure variations while gradually increasing and decreasing internal volume. Knowledge of distensibility was thus derived and then implemented with CMR data to test two applications. Firstly, a patient-specific compliant model of hypoplastic aorta suitable for connection in a mock circulatory loop for in-vitro tests was manufactured. Secondly, the right ventricular outflow tract (RVOT) of a patient necessitating pulmonary valve replacement was printed in order to physically test device insertion and assess patient's suitability for percutaneous pulmonary valve intervention. The distensibility of the material was identified in a range from 6.5 × 10(-3) mmHg(-1) for the 0.6 mm case, to 3.0 × 10(-3) mmHg(-1) for the 1.5 mm case. The models printed in the vertical orientation were always more compliant than their horizontal counterpart. Rapid prototyping of a compliant hypoplastic aorta and of a RVOT anatomical model were both feasible. Device insertion in the RVOT model was successful. Values of distensibility, compared with literature data, show that TangoPlus is suitable for manufacturing arterial phantoms, with

  10. Accelerated in vitro release testing of implantable PLGA microsphere/PVA hydrogel composite coatings.

    Science.gov (United States)

    Shen, Jie; Burgess, Diane J

    2012-01-17

    Dexamethasone loaded poly(lactic-co-glycolic acid) (PLGA) microsphere/PVA hydrogel composites have been investigated as an outer drug-eluting coating for implantable devices such as glucose sensors to counter negative tissue responses to implants. The objective of this study was to develop a discriminatory, accelerated in vitro release testing method for this drug-eluting coating using United States Pharmacopeia (USP) apparatus 4. Polymer degradation and drug release kinetics were investigated under "real-time" and accelerated conditions (i.e. extreme pH, hydro-alcoholic solutions and elevated temperatures). Compared to "real-time" conditions, the initial burst and lag phases were similar using hydro-alcoholic solutions and extreme pH conditions, while the secondary apparent zero-order release phase was slightly accelerated. Elevated temperatures resulted in a significant acceleration of dexamethasone release. The accelerated release data were able to predict "real-time" release when applying the Arrhenius equation. Microsphere batches with faster and slower release profiles were investigated under "real-time" and elevated temperature (60°C) conditions to determine the discriminatory ability of the method. The results demonstrated both the feasibility and the discriminatory ability of this USP apparatus 4 method for in vitro release testing of drug loaded PLGA microsphere/PVA hydrogel composites. This method may be appropriate for similar drug/device combination products and drug delivery systems. Copyright © 2011 Elsevier B.V. All rights reserved.

  11. Development of an in vitro Endotoxin Test for Monoolein–Water ...

    African Journals Online (AJOL)

    Purpose: Drugs that are administered by parenteral route must be apyrogenic. The aim of this study was to develop an in vitro endotoxin test for liquid crystalline gels for use as implants, using a monoolein–water liquid crystalline gel as a model. Methods: The gel-clot technique was used. The gel was dissolved first in ...

  12. An in vitro human skin test for assessing sensitization potential.

    Science.gov (United States)

    Ahmed, S S; Wang, X N; Fielding, M; Kerry, A; Dickinson, I; Munuswamy, R; Kimber, I; Dickinson, A M

    2016-05-01

    Sensitization to chemicals resulting in an allergy is an important health issue. The current gold-standard method for identification and characterization of skin-sensitizing chemicals was the mouse local lymph node assay (LLNA). However, for a number of reasons there has been an increasing imperative to develop alternative approaches to hazard identification that do not require the use of animals. Here we describe a human in-vitro skin explant test for identification of sensitization hazards and the assessment of relative skin sensitizing potency. This method measures histological damage in human skin as a readout of the immune response induced by the test material. Using this approach we have measured responses to 44 chemicals including skin sensitizers, pre/pro-haptens, respiratory sensitizers, non-sensitizing chemicals (including skin-irritants) and previously misclassified compounds. Based on comparisons with the LLNA, the skin explant test gave 95% specificity, 95% sensitivity, 95% concordance with a correlation coefficient of 0.9. The same specificity and sensitivity were achieved for comparison of results with published human sensitization data with a correlation coefficient of 0.91. The test also successfully identified nickel sulphate as a human skin sensitizer, which was misclassified as negative in the LLNA. In addition, sensitizers and non-sensitizers identified as positive or negative by the skin explant test have induced high/low T cell proliferation and IFNγ production, respectively. Collectively, the data suggests the human in-vitro skin explant test could provide the basis for a novel approach for characterization of the sensitizing activity as a first step in the risk assessment process. Copyright © 2015 John Wiley & Sons, Ltd.

  13. In vivo and in vitro testing for selenium and selenium compounds bioavailability assessment in foodstuff.

    Science.gov (United States)

    Moreda-Piñeiro, Jorge; Moreda-Piñeiro, Antonio; Bermejo-Barrera, Pilar

    2017-03-04

    The assessment of selenium and selenium species bioavailability in foodstuff is of special concern on the context of human nutrition. In vivo (human and animal), and in vitro tests are important approaches for estimating the bioavailability of toxic and essential compounds to humans. An overview on in vivo and in vitro bioavailability assays for releasing selenium and selenium species in foodstuffs is summarized. Se and Se species content in a foodstuff critically influence Se bioavailability and bioactivity to humans and animals. Se bioavailability is affected by foodstuff-matrix major composition and minor components. Foodstuffs processing and/or treatments could enhancement or decrease Se bioavailability. Experimental conditions such as the selection of healthy status of examined people (in in vivo humans approaches), the selection of animal model (in vivo animals approaches), or the selection of GI conditions (in in vitro tests) could determines the results. Thus, international standardized protocol for in vivo and in vitro approaches assessment is mandatory.

  14. THE ANTIGEN-SPECIFIC CELL IN VITRO TESTS FOR POST-VACCINATION ANTIPLAGUE IMMUNITY FORMATION

    Directory of Open Access Journals (Sweden)

    A. N. Kulichenko

    2017-01-01

    Full Text Available The possibility of post-vaccination anti-plague immunity evaluation was researched using antigen-stimulated cells tests in vitro and cytometry analysis. The object of study — the blood samples of 17 people immunised by the live plague vaccine (Yersinia pestis EV epicutaneously. Blood taking was carried out before vaccination and after immunisation on 7 and on 21 days, in 3 and in 6 months. Intensity antigen reactivity of lymphocytes was detected by cell tests in vitro, analysing markers of early (CD45+CD3+CD25+ and late (CD45+CD3+HLA-DR+ lymphocyte activation using flow cytometry. The complex of water-soluble Y. pestis antigens and allergen — pestin PP was tested as antigen. The high stimulating potential was defined of the water-soluble antigens Y. pestis complex. It is shown that coefficient of stimulation of relative level T- lymphocytes which express receptors for IL-2 was positive for all observation times after immunisation. The coefficient of stimulation had maximum values at 21 days (56.37% and at 3 (47.41% months. In identifying HLADR-positive lymphocytes before vaccination, the negative coefficient of stimulation was indicated on 7 and 21 days and the positive coefficient of stimulation was indicated at 3 and at 6 months. Analysis of intensity expression of early and late lymphocyte activation markers dynamics showed the possibility and prospect of application of cellular in vitro tests for the laboratory evaluation of specific reactivity of cellular immunity in both the early (7 days and late (6 months periods after vaccination. The results can be the basis for developing a new algorithm for assessment of immunological effectiveness of vaccination people against plague. It is the algorithm based on the identification of lymphocyte activation markers by antigen stimulation in conditions in vitro.

  15. Diagnosis of stinging insect allergy: utility of cellular in-vitro tests.

    Science.gov (United States)

    Scherer, Kathrin; Bircher, Andreas J; Heijnen, Ingmar Afm

    2009-08-01

    Diagnosis of stinging insect allergy is based on a detailed history, venom skin tests, and detection of venom-specific IgE. As an additional diagnostic tool, basophil responsiveness to venom allergens has been shown to be helpful in selected patients. This review summarizes the current diagnostic procedures for stinging insect allergy and discusses the latest developments in cellular in-vitro tests. Cellular assays have been evaluated in patients with Hymenoptera venom allergy. The diagnostic performance of the cellular mediator release test is similar to that of the flow cytometric basophil activation test (BAT), but the BAT has been the most intensively studied. BAT offers the possibility to assess basophil reactivity to allergens in their natural environment and to simultaneously analyze surface marker expression and intracellular signaling. It has been demonstrated that BAT represents a valuable additional diagnostic tool in selected patients when used in combination with other well established tests. A major limitation is the current lack of unified, standardized protocols. Flow cytometry offers huge possibilities to enhance knowledge of basophil functions. The BAT may be used as an additional test to confirm the diagnosis of stinging insect allergy in selected patients, provided that it is performed by an experienced laboratory using a validated assay. Test results have to be interpreted by clinicians familiar with the methodological aspects. The utility of the BAT to confirm allergy diagnosis and to predict the risk of subsequent systemic reactions may be improved by combined analysis of multiple surface markers and intracellular signaling pathways.

  16. Rapid lymphocyte immunoreactivity test utilizing [3H]uridine in vitro

    International Nuclear Information System (INIS)

    Pienkowski, M.M.; Lyerly, M.M.; Miller, H.C.

    1978-01-01

    A microculture assay utilizing [ 3 H]uridine incorporation was developed to test murine spleen lymphocyte immunoreactivity in vitro. Parameters of the culture technique which included cell density, doses of LPS, Con A, PHA, [ 3 H]uridine levels, and length of culture time were investigated. Responses were detectable at 4 h for all 3 mitogens, with labelling ranging up to 180% of the control value. By 8 h there was a 200-350% increase in mitogen-induced incorporation of radioactivity. Similar increases were observed in a serum-free system. The responses were the result of increased incorporation of label by stimulated cultures rather than decreased labeling of non-mitogen treated cultures over time. The [ 3 H]uridine incorporation was demonstrated to be the selective response of T or B cell populations when stimulated with appropriate lectins. This assay detects early RNA synthesis, as supported by experimental observations in which accumulation of radioactivity in stimulated lymphocytes was TCA precipitable, resistant to SDS treatment, and inhibited by actinomycin D. (Auth.)

  17. Biological evaluation of dental materials, in vitro and in vivo

    International Nuclear Information System (INIS)

    Kawahara, H.

    1982-01-01

    In this paper, the correlation between the user of tissue culture for in vitro tests and the tissue irritability and pupal response observed in in vitro tests, will be discussed. It would produce confusion if dental materials were standardised with the unreliable parameter of the living system in dynamic balance. Biological tests, both in vitro and in vivo, should be used for pre-standards testing, without any political control to establish physicochemical standards. As a first step, corrosion tests and the dissolution dosje of toxic components from the material in the tissue culture medium and/or artificial salvia should be standardised under conditions simulating the oral environment. The CNC method and photo-pattern analysis are used for the interpretation of cytotoxicity. The need for biological testing, both in vitro and in vivo, definitely exists in order to obtain physicochemical standards, with a biological simulation depending upon the feedback obtained from the results of in vitro and in vivo tests

  18. Influence of different test parameters on in vitro drug release from topical diclofenac formulations in a vertical diffusion cell setup.

    Science.gov (United States)

    Klein, S

    2013-07-01

    In the past decades, the vertical diffusion cell has emerged as a useful device for testing drug release of topical dosage forms. However, to date neither a general USP method nor formulation-related monographs have been published in international pharmacopoeia. The purpose of the present work was to examine the influence of different test parameters in a vertical diffusion cell setup on in vitro drug release from semi-solid preparations for cutaneous application. Diclofenac was selected as the model compound. Release experiments were performed in a 7 ml Microett vertical diffusion cell system. Various test parameters, including the media composition and pH, degassing, membrane material and pore size, stirring speed and stirrer type, were varied. Results obtained with different test parameter settings clearly indicate that both drug properties and instrumental details can have a huge impact on the outcome of in vitro diffusion/drug release studies with the vertical diffusion cell. Thus, the selection of adequate test parameters is crucial for the success of the release experiments and, as shown in the present study, optimal test parameters/conditions need to be established and validated on a case by case study.

  19. Deciphering defective amelogenesis using in vitro culture systems.

    Science.gov (United States)

    Arinawati, Dian Yosi; Miyoshi, Keiko; Tanimura, Ayako; Horiguchi, Taigo; Hagita, Hiroko; Noma, Takafumi

    2018-04-01

    The conventional two-dimensional (2D) in vitro culture system is frequently used to analyze the gene expression with or without extracellular signals. However, the cells derived from primary culture and cell lines frequently deviate the gene expression profile compared to the corresponding in vivo samples, which sometimes misleads the actual gene regulation in vivo. To overcome this gap, we developed the comparative 2D and 3D in vitro culture systems and applied them to the genetic study of amelogenesis imperfecta (AI) as a model. Recently, we found specificity protein 6 (Sp6) mutation in an autosomal-recessive AI rat that was previously named AMI. We constructed 3D structure of ARE-B30 cells (AMI-derived rat dental epithelial cells) or G5 (control wild type cells) combined with RPC-C2A cells (rat pulp cell line) separated by the collagen membrane, while in 2D structure, ARE-B30 or G5 was cultured with or without the collagen membrane. Comparative analysis of amelogenesis-related gene expression in ARE-B30 and G5 using our 2D and 3D in vitro systems revealed distinct expression profiles, showing the causative outcomes. Bone morphogenetic protein 2 and follistatin were reciprocally expressed in G5, but not in ARE-B30 cells. All-or-none expression of amelotin, kallikrein-related peptidase 4, and nerve growth factor receptor was observed in both cell types. In conclusion, our in vitro culture systems detected the phenotypical differences in the expression of the stage-specific amelogenesis-related genes. Parallel analysis with 2D and 3D culture systems may provide a platform to understand the molecular basis for defective amelogenesis caused by Sp6 mutation. Copyright © 2017 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  20. In vitro characterization of microcontainers as an oral drug delivery system

    DEFF Research Database (Denmark)

    Nielsen, Line Hagner; Keller, Stephan Sylvest; Jacobsen, J.

    We here present in vitro studies showing the promise of microcontainers (fabricated in either SU-8 or Poly(lactic acid) (PLLA)) as an oral drug delivery system for the poorly watersoluble drug, furosemide.......We here present in vitro studies showing the promise of microcontainers (fabricated in either SU-8 or Poly(lactic acid) (PLLA)) as an oral drug delivery system for the poorly watersoluble drug, furosemide....

  1. Neuroblast of the grasshopper embryo as a new mutagen test system. Pt. 1. In vitro radiosensitivity

    Energy Technology Data Exchange (ETDEWEB)

    Liang, J C; Gaulden, M E [Texas Univ., Dallas (USA). Dept. of Radiology

    1982-04-01

    The neuroblasts of the grasshopper embryo (Chortophaga viridifasciata De Geer) are being studied to determine their suitability for detecting environmental clastogens (chromosome-breaking agents). They are very sensitive to the induction of chromosome breakage by radiation in vitro. Their sensitvity, 0.011 break/cell/R, is 4-5 times higher than pollen mother cells of Tradescantia (micronuclei), 10 times higher than either human lymphocytes or Chinese hamster cells (metaphase chromosome aberrations), and 15 times higher than mouse erythroblasts (micronuclei). Furthermore, they have no spontaneous chromosome breakage, which facilitates the detection of agents that break chromosomes. The present study shows that Chortophaga embryos maintain normal mitotic activity in vitro for 5 cell cycles at 38/sup 0/C (20 h), and that neuroblasts of embryos grown in vitro have the same radiosensitivity as those of embryos in vivo. Thus in vitro exposure of grasshopper embryos is a promising method for obtaining data on the response of neuroblasts to chemical clastogens.

  2. Possibilities for in-vitro testing of controlled-release parenterals.

    OpenAIRE

    Kožák, Jan

    2014-01-01

    The objective of this study was to start with the development of a gel-based medium simulating intramuscular environment for in-vitro drug release testing as an alternative to currently used water-based media. A pork meat was selected as a model of the muscle tissue. At the first part of the study, diffusion rate was proposed as the most determining factor of drug release in gel-based medium. Consequently, the diffusion rate of sodium fluorescein in pork meat was compared with the diffusion r...

  3. Etest cannot be recommended for in vitro susceptibility testing of mucorales.

    Science.gov (United States)

    Caramalho, Rita; Maurer, Elisabeth; Binder, Ulrike; Araújo, Ricardo; Dolatabadi, Somayeh; Lass-Flörl, Cornelia; Lackner, Michaela

    2015-01-01

    Amphotericin B and posaconazole susceptibility patterns were determined for the most prevalent Mucorales, following EUCAST (European Committee on Antimicrobial Susceptibility Testing) broth microdilution guidelines. In parallel, Etest was performed and evaluated against EUCAST. The overall agreement of MICs gained with Etest and EUCAST was 75.1%; therefore, Etest cannot be recommended for antifungal susceptibility testing of Mucorales. Amphotericin B was the most active drug against Mucorales species in vitro, while the activities of posaconazole were more restricted. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  4. A new in vitro screening system for anticancer drugs for the treatment of non-small cell lung cancer

    International Nuclear Information System (INIS)

    Hanauske, U.; Hanauske, A.R.; Clark, G.M.; Tsen, D.; Buchok, J.; Hoff, D.D. von

    1989-01-01

    We have evaluated a semiautomated radiometric assay (BACTEC 460 system) for screening of activity of anticancer drugs against human non-small cell lung cancer cell lines. Cells from seven cell lines were exposed to standard antineoplastic agents at four different concentrations using a 1-h incubation. Alpha 2-interferon was tested using a continuous incubation. In vitro drug activity was analyzed as a function of the clinically achievable serum concentration. Our results indicate that two cell lines (CALU-3, SK-MES-1) exhibit in vitro drug sensitivity patterns closest to those observed in clinical studies. These two cell lines might therefore be most useful for screening new anticancer compounds for activity against non-small cell lung cancer. The radiometric assay is a semiautomated system which has advantages over other, more time-consuming screening systems

  5. In Vitro Screening of Environmental Chemicals for Targeted Testing Prioritization: The ToxCast Project

    OpenAIRE

    Judson, Richard S.; Houck, Keith A.; Kavlock, Robert J.; Knudsen, Thomas B.; Martin, Matthew T.; Mortensen, Holly M.; Reif, David M.; Rotroff, Daniel M.; Shah, Imran; Richard, Ann M.; Dix, David J.

    2009-01-01

    Background Chemical toxicity testing is being transformed by advances in biology and computer modeling, concerns over animal use, and the thousands of environmental chemicals lacking toxicity data. The U.S. Environmental Protection Agency?s ToxCast program aims to address these concerns by screening and prioritizing chemicals for potential human toxicity using in vitro assays and in silico approaches. Objectives This project aims to evaluate the use of in vitro assays for understanding the ty...

  6. In vitro screening of environmental chemicals for targeted testing prioritization: the ToxCast project.

    Science.gov (United States)

    Judson, Richard S; Houck, Keith A; Kavlock, Robert J; Knudsen, Thomas B; Martin, Matthew T; Mortensen, Holly M; Reif, David M; Rotroff, Daniel M; Shah, Imran; Richard, Ann M; Dix, David J

    2010-04-01

    Chemical toxicity testing is being transformed by advances in biology and computer modeling, concerns over animal use, and the thousands of environmental chemicals lacking toxicity data. The U.S. Environmental Protection Agency's ToxCast program aims to address these concerns by screening and prioritizing chemicals for potential human toxicity using in vitro assays and in silico approaches. This project aims to evaluate the use of in vitro assays for understanding the types of molecular and pathway perturbations caused by environmental chemicals and to build initial prioritization models of in vivo toxicity. We tested 309 mostly pesticide active chemicals in 467 assays across nine technologies, including high-throughput cell-free assays and cell-based assays, in multiple human primary cells and cell lines plus rat primary hepatocytes. Both individual and composite scores for effects on genes and pathways were analyzed. Chemicals displayed a broad spectrum of activity at the molecular and pathway levels. We saw many expected interactions, including endocrine and xenobiotic metabolism enzyme activity. Chemicals ranged in promiscuity across pathways, from no activity to affecting dozens of pathways. We found a statistically significant inverse association between the number of pathways perturbed by a chemical at low in vitro concentrations and the lowest in vivo dose at which a chemical causes toxicity. We also found associations between a small set of in vitro assays and rodent liver lesion formation. This approach promises to provide meaningful data on the thousands of untested environmental chemicals and to guide targeted testing of environmental contaminants.

  7. Absorbable magnesium-based stent: physiological factors to consider for in vitro degradation assessments

    OpenAIRE

    Wang, Juan; Smith, Christopher E.; Sankar, Jagannathan; Yun, Yeoheung; Huang, Nan

    2015-01-01

    Absorbable metals have been widely tested in various in vitro settings using cells to evaluate their possible suitability as an implant material. However, there exists a gap between in vivo and in vitro test results for absorbable materials. A lot of traditional in vitro assessments for permanent materials are no longer applicable to absorbable metallic implants. A key step is to identify and test the relevant microenvironment and parameters in test systems, which should be adapted according ...

  8. In vitro antibacterial activity of adhesive systems on Streptococcus mutans.

    Science.gov (United States)

    Paradella, Thaís Cachuté; Koga-Ito, Cristiane Yumi; Jorge, Antonio Olavo Cardoso

    2009-04-01

    To evaluate the antibacterial activity of three adhesive systems -- Prime & Bond 2.1 (PB), Clearfil SE Bond (CS) and One Up Bond F (OU) -- on Streptococcus mutans in vitro. Adherence and agar disk-diffusion tests were performed. For the adherence testing, 40 human enamel specimens (4 mm2) were sterilized and the adhesive sytems were applied (n = 10). The control group did not receive the application of any adhesive system. Specimens were immersed in brain heart infusion broth (BHI) inoculated with S. mutans standardized suspension (10(6) cells/ml) for 48 h at 37 degrees C and 5% CO2. The number of S. mutans cells adhered to each specimen was evaluated by the plating method on BHI agar. For agar disk-diffusion testing, adhesive disks and disks soaked in distilled water (negative control) or 0.2% chlorexidine (positive control) were incubated with S. mutans for 48 h. The diameters of the zones of bacterial inhibition were measured. Adherence data were transformed in logarithms of base 10 (log10). Data were submitted to Kruskal-Wallis and Student-Neuman-Keuls tests at the 5% level of significance. The results of the adherence test showed that One Up Bond F (OU) and Clearfil SE Bond (CS) did not differ significantly from one another, but allowed significantly less adherence than Prime & Bond 2.1 (PB) and control [mean log10 (standard deviation) values: PB 6.10 (0.19); CS primer 4.55 (0.98); OU 4.65 (0.54); control group 6.34 (0.27)]. The disk-diffusion test showed no significant difference between OU (diameter in mm: 3.02 +/- 0.13) and CS (3.0 +/- 0.12), but both were significantly more effective in inhibiting bacterial growth than PB (1.0 +/- 0.10). The self-etching systems Clearfil SE Bond and One Up Bond F presented a greater inhibitory effect against S. mutans, also in terms of adherence, than did the conventional system, Prime & Bond 2.1.

  9. Study of the system of tuberous root induction in vitro from ...

    African Journals Online (AJOL)

    Abstract. This study investigated the induction system of tuberous root in vitro from Rehmannia glutinosa. The roles of plant growth substance, carbohydrates, and minerals were evaluated for induction and development of tuberous root in vitro. The results show that Murashige and Skoog (MS) contributed greatly to induction ...

  10. 10 CFR 31.11 - General license for use of byproduct material for certain in vitro clinical or laboratory testing.

    Science.gov (United States)

    2010-01-01

    ... therefrom, to human beings or animals. (2) Iodine-131, in units not exceeding 10 microcuries each for use in... microcuries each for use in in vitro clinical or laboratory tests not involving internal or external... (tritium), in units not exceeding 50 microcuries each for use in in vitro clinical or laboratory tests not...

  11. Recommendations for In Vitro and In Vivo Testing of Magnetic Nanoparticle Hyperthermia Combined with Radiation Therapy

    Directory of Open Access Journals (Sweden)

    Spiridon V. Spirou

    2018-05-01

    Full Text Available Magnetic nanoparticle (MNP-mediated hyperthermia (MH coupled with radiation therapy (RT is a novel approach that has the potential to overcome various practical difficulties encountered in cancer treatment. In this work, we present recommendations for the in vitro and in vivo testing and application of the two treatment techniques. These recommendations were developed by the members of Working Group 3 of COST Action TD 1402: Multifunctional Nanoparticles for Magnetic Hyperthermia and Indirect Radiation Therapy (“Radiomag”. The purpose of the recommendations is not to provide definitive answers and directions but, rather, to outline those tests and considerations that a researcher must address in order to perform in vitro and in vivo studies. The recommendations are divided into 5 parts: (a in vitro evaluation of MNPs; (b in vitro evaluation of MNP-cell interactions; (c in vivo evaluation of the MNPs; (d MH combined with RT; and (e pharmacokinetic studies of MNPs. Synthesis and characterization of the MNPs, as well as RT protocols, are beyond the scope of this work.

  12. Magnesium alloys: predicting in vivo corrosion with in vitro immersion testing.

    Science.gov (United States)

    Walker, Jemimah; Shadanbaz, Shaylin; Kirkland, Nicholas T; Stace, Edward; Woodfield, Tim; Staiger, Mark P; Dias, George J

    2012-05-01

    Magnesium (Mg) and its alloys have been proposed as degradable replacements to commonly used orthopedic biomaterials such as titanium alloys and stainless steel. However, the corrosion of Mg in a physiological environment remains a difficult characteristic to accurately assess with in vitro methods. The aim of this study was to identify a simple in vitro immersion test that could provide corrosion rates similar to those observed in vivo. Pure Mg and five alloys (AZ31, Mg-0.8Ca, Mg-1Zn, Mg-1Mn, Mg-1.34Ca-3Zn) were immersed in either Earle's balanced salt solution (EBSS), minimum essential medium (MEM), or MEM-containing 40 g/L bovine serum albumin (MEMp) for 7, 14, or 21 days before removal and assessment of corrosion by weight loss. This in vitro data was compared to in vivo corrosion rates of the same materials implanted in a subcutaneous environment in Lewis rats for equivalent time points. The results suggested that, for the alloys investigated, the EBSS buffered with sodium bicarbonate provides a rate of degradation comparable to those observed in vivo. In contrast, the addition of components such as (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) (HEPES), vitamins, amino acids, and albumin significantly increased corrosion rates. Based on these findings, it is proposed that with this in vitro protocol, immersion of Mg alloys in EBSS can be used as a predictor of in vivo corrosion. Copyright © 2012 Wiley Periodicals, Inc.

  13. A novel physiological testing device to study knee biomechanics in vitro

    NARCIS (Netherlands)

    van de Bunt, Fabian; Emanuel, Kaj S.; Wijffels, Thomas; Kooren, Peter N.; Kingma, Idsart; Smit, Theodoor H.

    2017-01-01

    Background To properly study knee kinetics, kinematics and the effects of injury and surgical treatment in vitro, the knee should be constrained as little as possible, while imposing physiological loads. A novel dynamic biomechanical knee system (BKS) is presented here. The aim of this study was to

  14. A novel physiological testing device to study knee biomechanics in vitro

    NARCIS (Netherlands)

    van de Bunt, Fabian; Emanuel, Kaj S.; Wijffels, Thomas; Kooren, Peter N.; Kingma, Idsart; Smit, Theodoor H.

    2017-01-01

    Background: To properly study knee kinetics, kinematics and the effects of injury and surgical treatment in vitro, the knee should be constrained as little as possible, while imposing physiological loads. A novel dynamic biomechanical knee system (BKS) is presented here. The aim of this study was to

  15. In vitro dissolution of curium oxide using a phagolysosomal simulant solvent system

    International Nuclear Information System (INIS)

    Helfinstine, S.Y.; Guilmette, R.A.; Schlapper, G.A.

    1992-01-01

    Detailed study of actinide oxide behavior in alveolar macrophages (AM) in vitro is limited because of the short life span of these cells in culture. We created an in vitro dissolution system that could mimic the acidic phagolysosomal environment for the actinide and be maintained for an indefinite period so that dissolution of more insoluble materials could be measured. The dissolution system for this investigation, consisting of nine different solutions of HCl and the chelating agent diethylenetriamine pentaacetate (DTPA) in distilled water, is called the phagolysosomal simulant solvent (PSS). In this system, both the pH and the amount of DTPA were varied. We could observe the effect of altering pH within a range of 4.0-6.0 (similar to that of the phagolysosome) and the effect of the molar ratio of DTPA to curium at 1000: 1, 100;1, or 10:1. We chose curium sesquioxide ( 244 Cm 2 O 3 ) to validate the PSS for actinide dissolution versus that occurring in AM in vitro because it dissolves significantly in less than 1 week. The polydisperse 244 Cm 2 O 3 aerosol was generated, collected on filters, resuspended, and added to the PSS solutions and to cultured canine AM. By comparing dissolution in the two systems directly, we hoped to arrive at an optimum PSS for future dissolution studies. PSS and cell culture samples were taken daily for 7 days after exposure and tested for the solubilized curium. The amount of soluble material was determined by ultracentrifugation to separate the insoluble CM 2 O 3 from the soluble curium in the PSS solutions and filtration for the cell-containing material. After separating the soluble and insoluble fractions, the samples were analyzed using alpha liquid scintillation counting. Time-dependent dissolution measurements from the PSS/AM showed that the CM 2 O 3 dissolution was similar for both the PSS solutions and the cultured AM. 13 refs., 4 tabs

  16. Test systems to identify reproductive toxicants.

    Science.gov (United States)

    Riecke, K; Stahlmann, R

    2000-09-01

    Experience with drugs and other xenobiotics indicates that both animal testing and epidemiological studies are necessary to provide adequate data for an estimation of risks that might be associated with exposure to a chemical substance. In this review, the pros and cons of test systems for reproductive toxicity are discussed. Usually, several studies are performed to cover the different phases of the reproductive cycle. In the preclinical development of drugs, the three so-called 'segment testing protocols' have been used for several decades now. More recently, new testing concepts have been accepted internationally which include more flexibility in implementation. Several examples of compounds with the potential for reproductive toxicity are presented in more detail in a discussion of some pitfalls of the tests for fertility (phthalates and fluoroquinolones), teratogenicity (acyclovir and protease inhibitors) and postnatal developmental toxicity (fluoroquinolones). In addition, important aspects of kinetics and metabolism as a prerequisite for a rational interpretation of results from toxicological studies are briefly discussed. In vitro assays are useful for supplementing the routinely used in vivo approaches or for studying an expected or defined effect, but they are not suitable for revealing an unknown effect of a chemical on the complex reproductive process.

  17. In vivo and in vitro experimental study on cervix cancer with combination of HSV-TK/GCV suicide gene therapy system and 60Co radiotherapy

    International Nuclear Information System (INIS)

    Chen Daozhen; Xue Wenqun; Zhan Huiying; Zhu Yunxia; Yang Youyi; Liu Lu; Tang Qiusha

    2006-01-01

    Objective: To evaluate the killing effect of HSV-TK/GCV suicide gene therapy system combined with 60 Co radiotherapy on human cervical cancer HeLa cell line in vivo and in vitro, and to explore radiosensitization by the HSV-TK/GCV system. Methods: The HSV-TK/GCV suicide gene therapy system and 60 Co radiotherapy were used separately or in combination for human cervical cancer HeLa cell line in vivo and in vitro to compare their effects. Colony formation test and the rate of radiosensitization effect(E/O) were employed to observed the radiosensitization by the HSV-TK/GCV system. Results: The HSV-TK/GCV suicide gene therapy system showed strong therapeutic effects on HeLa cells both in vitro and in vivo (the inhibition rates were 45.8% and 39.5%, respectively). Moreover, the combined application of gene therapy and radiotherapy exhibited stronger therapeutic effects in vitro and in vivo (the inhibition rate was 87.5% in vitro, and was 87.9% in vivo) (P 1.4), indicating the HSV-TK/GCV system could exert a sensitizing effect on 60 Co radiotherapy of the transplanted human cervical cancer cells in nude mice. Conclusion: The HSV-TK/GCV system has radiosensitizationaction. Gene therapy combined with radiotherapy may be a good supplementary method for synthetic treatment of cervical cancer. (authors)

  18. Assessment of an in vitro whole cigarette smoke exposure system: The Borgwaldt RM20S 8-syringe smoking machine

    Directory of Open Access Journals (Sweden)

    McAughey John

    2011-08-01

    Full Text Available Abstract Background There have been many recent developments of in vitro cigarette smoke systems closely replicating in vivo exposures. The Borgwaldt RM20S smoking machine (RM20S enables the serial dilution and delivery of cigarette smoke to exposure chambers for in vitro analyses. In this study we have demonstrated reliability and robustness testing of the RM20S in delivering smoke to in vitro cultures using an in-house designed whole smoke exposure chamber. Results The syringe precision and accuracy of smoke dose generated by the RM20S was assessed using a methane gas standard and resulted in a repeatability error of ≤9%. Differential electrical mobility particle spectrometry (DMS measured smoke particles generated from reference 3R4F cigarettes at points along the RM20S. 53% ± 5.9% of particles by mass reached the chamber, the remainder deposited in the syringe or connecting tubing and ~16% deposited in the chamber. Spectrofluorometric quantification of particle deposition within chambers indicated a positive correlation between smoke concentration and particle deposition. In vitro air-liquid interface (ALI cultures (H292 lung epithelial cells, exposed to whole smoke (1:60 dilution (smoke:air, equivalent to ~5 μg/cm2 demonstrated uniform smoke delivery within the chamber. Conclusions These results suggest this smoke exposure system is a reliable and repeatable method of generating and exposing ALI in vitro cultures to cigarette smoke. This system will enable the evaluation of future tobacco products and individual components of cigarette smoke and may be used as an alternative in vitro tool for evaluating other aerosols and gaseous mixtures such as air pollutants, inhaled pharmaceuticals and cosmetics.

  19. Energy Systems Test Area (ESTA). Power Systems Test Facilities

    Science.gov (United States)

    Situ, Cindy H.

    2010-01-01

    This viewgraph presentation provides a detailed description of the Johnson Space Center's Power Systems Facility located in the Energy Systems Test Area (ESTA). Facilities and the resources used to support power and battery systems testing are also shown. The contents include: 1) Power Testing; 2) Power Test Equipment Capabilities Summary; 3) Source/Load; 4) Battery Facilities; 5) Battery Test Equipment Capabilities Summary; 6) Battery Testing; 7) Performance Test Equipment; 8) Battery Test Environments; 9) Battery Abuse Chambers; 10) Battery Abuse Capabilities; and 11) Battery Test Area Resources.

  20. In vitro comparison of Günther Tulip and Celect filters: testing filtering efficiency and pressure drop.

    Science.gov (United States)

    Nicolas, M; Malvé, M; Peña, E; Martínez, M A; Leask, R

    2015-02-05

    In this study, the trapping ability of the Günther Tulip and Celect inferior vena cava filters was evaluated. Thrombus capture rates of the filters were tested in vitro in horizontal position with thrombus diameters of 3 and 6mm and tube diameter of 19mm. The filters were tested in centered and tilted positions. Sets of 30 clots were injected into the model and the same process was repeated 20 times for each different condition simulated. Pressure drop experienced along the system was also measured and the percentage of clots captured was recorded. The Günther Tulip filter showed superiority in all cases, trapping almost 100% of 6mm clots both in an eccentric and tilted position and trapping 81.7% of the 3mm clots in a centered position and 69.3% in a maximum tilted position. The efficiency of all filters tested decreased as the size of the embolus decreased and as the filter was tilted. The injection of 6 clots raised the pressure drop to 4.1mmHg, which is a reasonable value that does not cause the obstruction of blood flow through the system. Copyright © 2014 Elsevier Ltd. All rights reserved.

  1. Extreme testing of undiluted e-cigarette aerosol in vitro using an Ames air-agar-interface technique.

    Science.gov (United States)

    Thorne, D; Hollings, M; Seymour, A; Adamson, J; Dalrymple, A; Ballantyne, M; Gaca, M

    2018-04-01

    There is a growing consensus that e-cigarettes hold the potential for reducing the harm associated with cigarette smoking. Recently published studies have reported in vitro testing of e-cigarettes, demonstrating reduced toxicological and biological effects. Few studies however have reported the use of e-cigarettes under extreme testing conditions. To assess the full mutagenic potential of a commercially available electronic-cigarette (Vype ePen), this study investigated the delivery of aerosol under extreme conditions, using a scaled-down 35 mm plate Ames bacterial reverse mutagenicity assay. S. typhimurium strains TA98, TA100, TA97, TA104 and E. coli WP2 uvrA pKM101 with or without metabolic activation (S9), were employed. Using a modified Vitrocell VC 10 exposure system 0, 180, 360, 540, 720 or 900 puffs of undiluted e-cigarette aerosol was generated and delivered to bacterial cultures aligned to reported human consumption data. The results demonstrate that no mutagenic activity was observed in any strain under any test condition even when exposed to 900 puffs of undiluted e-cigarette aerosols +/- S9. Positive control responses were observed in all strains +/- S9. Nicotine assessments demonstrated an increased and consistent aerosol delivery, with calculated maximum doses of ∼1 mg/mL delivery of nicotine. These data demonstrate the validity of this unique testing approach and adds further information to the growing weight of evidence that e-cigarettes offer substantially reduced exposure when compared to conventional cigarette smoke. For future in vitro assessments of next generation tobacco and nicotine products, the generation, delivery and testing of undiluted aerosols can now be considered. Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.

  2. In vitro and in vivo tests of PLA/d-HAp nanocomposite

    Science.gov (United States)

    Thom Nguyen, Thi; Hoang, Thai; Mao Can, Van; Son Ho, Anh; Hai Nguyen, Song; Thu Trang Nguyen, Thi; Pham, Thi Nam; Phuong Nguyen, Thu; Le Hien Nguyen, Thi; Thanh Dinh Thi, Mai

    2017-12-01

    The bioactivity of the PLA/d-HAp nanocomposite with 30 wt.% d-HAp was evaluated by in vitro tests and indicated that after 7 immersion days in SBF solution, PLA amorphous part was hydrolyzed and PLA crystal part was remained. The formation of apatite on the surface of the material was observed. The in vivo test results of PLA/d-HAp nanocomposite (70/30 wt/wt) on femur of dogs displayed that 3 months after grafting, the materials did not induce any osteitis, osteomyelitis or structural abnormalities. The histological and x-ray image demonstrated a growth of the bone into the material area, while osteitis and osteomyelitis were not observed.

  3. Biology-inspired Microphysiological System Approaches to Solve the Prediction Dilemma of Substance Testing

    Science.gov (United States)

    Marx, Uwe; Andersson, Tommy B.; Bahinski, Anthony; Beilmann, Mario; Beken, Sonja; Cassee, Flemming R.; Cirit, Murat; Daneshian, Mardas; Fitzpatrick, Susan; Frey, Olivier; Gaertner, Claudia; Giese, Christoph; Griffith, Linda; Hartung, Thomas; Heringa, Minne B.; Hoeng, Julia; de Jong, Wim H.; Kojima, Hajime; Kuehnl, Jochen; Luch, Andreas; Maschmeyer, Ilka; Sakharov, Dmitry; Sips, Adrienne J. A. M.; Steger-Hartmann, Thomas; Tagle, Danilo A.; Tonevitsky, Alexander; Tralau, Tewes; Tsyb, Sergej; van de Stolpe, Anja; Vandebriel, Rob; Vulto, Paul; Wang, Jufeng; Wiest, Joachim; Rodenburg, Marleen; Roth, Adrian

    2017-01-01

    Summary The recent advent of microphysiological systems – microfluidic biomimetic devices that aspire to emulate the biology of human tissues, organs and circulation in vitro – is envisaged to enable a global paradigm shift in drug development. An extraordinary US governmental initiative and various dedicated research programs in Europe and Asia have led recently to the first cutting-edge achievements of human single-organ and multi-organ engineering based on microphysiological systems. The expectation is that test systems established on this basis would model various disease stages, and predict toxicity, immunogenicity, ADME profiles and treatment efficacy prior to clinical testing. Consequently, this technology could significantly affect the way drug substances are developed in the future. Furthermore, microphysiological system-based assays may revolutionize our current global programs of prioritization of hazard characterization for any new substances to be used, for example, in agriculture, food, ecosystems or cosmetics, thus, replacing laboratory animal models used currently. Thirty-five experts from academia, industry and regulatory bodies present here the results of an intensive workshop (held in June 2015, Berlin, Germany). They review the status quo of microphysiological systems available today against industry needs, and assess the broad variety of approaches with fit-for-purpose potential in the drug development cycle. Feasible technical solutions to reach the next levels of human biology in vitro are proposed. Furthermore, key organ-on-a-chip case studies, as well as various national and international programs are highlighted. Finally, a roadmap into the future is outlined, to allow for more predictive and regulatory-accepted substance testing on a global scale. PMID:27180100

  4. Long-term in-vitro precision of direct digital X-ray radiogrammetry

    International Nuclear Information System (INIS)

    Dhainaut, Alvilde; Hoff, Mari; Kaelvesten, Johan; Lydersen, Stian; Forslind, Kristina; Haugeberg, Glenn

    2011-01-01

    Digital X-ray radiogrammetry (DXR) calculates peripheral bone mineral density (BMD) from hand radiographs. The short-term precision for direct DXR has been reported to be highly satisfactory. However, long-term precision for this method has not been examined. Thus, the aim of this study was to examine the long-term in-vitro precision for the new direct digital version of DXR. The in-vitro precision for direct DXR was tested with cadaver phantoms on four different X-ray systems at baseline, 3 months, 6 months, and in one machine also at 12 months. At each time point, 31 measurements were performed. The in-vitro longitudinal precision for the four radiographic systems ranged from 0.22 to 0.43% expressed as coefficient of variation (CV%). The smallest detectable difference (SDD) ranged from 0.0034 to 0.0054 g/cm 2 . The in vitro long-term precision for direct DXR was comparable to the previous reported short-term in-vitro precision for all tested X-ray systems. These data show that DXR is a stable method for detecting small changes in bone density during 6-12 months of follow-up. (orig.)

  5. Critical review of the current and future challenges associated with advanced in vitro systems towards the study of nanoparticle (secondary) genotoxicity.

    Science.gov (United States)

    Evans, Stephen J; Clift, Martin J D; Singh, Neenu; de Oliveira Mallia, Jefferson; Burgum, Michael; Wills, John W; Wilkinson, Thomas S; Jenkins, Gareth J S; Doak, Shareen H

    2017-01-01

    With the need to understand the potential biological impact of the plethora of nanoparticles (NPs) being manufactured for a wide range of potential human applications, due to their inevitable human exposure, research activities in the field of NP toxicology has grown exponentially over the last decade. Whilst such increased research efforts have elucidated an increasingly significant knowledge base pertaining to the potential human health hazard posed by NPs, understanding regarding the possibility for NPs to elicit genotoxicity is limited. In vivo models are unable to adequately discriminate between the specific modes of action associated with the onset of genotoxicity. Additionally, in line with the recent European directives, there is an inherent need to move away from invasive animal testing strategies. Thus, in vitro systems are an important tool for expanding our mechanistic insight into NP genotoxicity. Yet uncertainty remains concerning their validity and specificity for this purpose due to the unique challenges presented when correlating NP behaviour in vitro and in vivo This review therefore highlights the current state of the art in advanced in vitro systems and their specific advantages and disadvantages from a NP genotoxicity testing perspective. Key indicators will be given related to how these systems might be used or improved to enhance understanding of NP genotoxicity. © The Author 2016. Published by Oxford University Press on behalf of the UK Environmental Mutagen Society.

  6. Medical chilling device designed for hypothermic hydration graft storage system: Design, thermohydrodynamic modeling, and preliminary testing

    Energy Technology Data Exchange (ETDEWEB)

    Seo, Jung Hwan [Hongik University, Seoul (Korea, Republic of)

    2015-02-15

    Hypothermic hydration graft storage is essential to reduce the metabolic demand of cells in vitro. The alleviated metabolic demands reduce the emergence rate of anaerobic metabolism generating adenosine triphosphate (ATP) energy that creates free radicals. The cessive free radicals can damage cells and tissues due to their highly oxidative power with molecules. Current cooling systems such as a conventional air cooling system and an ice pack system are inappropriate for chilling cell tissues in vitro because of inconvenience in use and inconsistent temperature sustainability caused by large size and progressive melting, respectively. Here, we develop a medical chilling device (MCD) for hypothermic hydration graft storage based on thermo-hydrodynamic modeling and thermal electric cooling technology. Our analysis of obtained hydrodynamic thermal behavior of the MCD revealed that the hypothermic condition of 4 .deg. C was continuously maintained, which increased the survival rates of cells in vitro test by reduced free radicals. The validated performance of the MCD promises future development of an optimal hypothermic hydration graft storage system designed for clinical use.

  7. Accelerated in vitro release testing method for naltrexone loaded PLGA microspheres.

    Science.gov (United States)

    Andhariya, Janki V; Choi, Stephanie; Wang, Yan; Zou, Yuan; Burgess, Diane J; Shen, Jie

    2017-03-30

    The objective of the present study was to develop a discriminatory and reproducible accelerated release testing method for naltrexone loaded parenteral polymeric microspheres. The commercially available naltrexone microsphere product (Vivitrol ® ) was used as the testing formulation in the in vitro release method development, and both sample-and-separate and USP apparatus 4 methods were investigated. Following an in vitro drug stability study, frequent media replacement and addition of anti-oxidant in the release medium were used to prevent degradation of naltrexone during release testing at "real-time" (37°C) and "accelerated" (45°C), respectively. The USP apparatus 4 method was more reproducible than the sample-and-separate method. In addition, the accelerated release profile obtained using USP apparatus 4 had a shortened release duration (within seven days), and good correlation with the "real-time" release profile. Lastly, the discriminatory ability of the developed accelerated release method was assessed using compositionally equivalent naltrexone microspheres with different release characteristics. The developed accelerated USP apparatus 4 release method was able to detect differences in the release characteristics of the prepared naltrexone microspheres. Moreover, a linear correlation was observed between the "real-time" and accelerated release profiles of all the formulations investigated, suggesting that the release mechanism(s) may be similar under both conditions. These results indicate that the developed accelerated USP apparatus 4 method has the potential to be an appropriate fast quality control tool for long-acting naltrexone PLGA microspheres. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. ACETURATO DE DIMINAZENO LIPOSSOMAL NO TRATAMENTO DA INFECÇÃO POR Trypanosoma evansi: TESTES in vitro E in vivo

    OpenAIRE

    Camila Belmonte Oliveira

    2014-01-01

    Este estudo teve como objetivo desenvolver e testar lipossomas de aceturato de diminazeno em testes in vitro e in vivo visando o controle de Trypanosoma evansi. O teste in vitro foi realizado em meio de cultura nas concentrações de 0,25, 0,5, 1, 2 e 3 μg/mL de aceturato de diminazeno convencional (C-DMZ) e lipossomal (L-DMZ). Para os testes in vivo foram utilizados 114 ratos (Rattus norvegicus) divididos em seis grupos (A, B, C, D, E e F) em dois experimentos, um para aval...

  9. Correlation of In Vivo Versus In Vitro Benchmark Doses (BMDs) Derived From Micronucleus Test Data: A Proof of Concept Study.

    Science.gov (United States)

    Soeteman-Hernández, Lya G; Fellows, Mick D; Johnson, George E; Slob, Wout

    2015-12-01

    In this study, we explored the applicability of using in vitro micronucleus (MN) data from human lymphoblastoid TK6 cells to derive in vivo genotoxicity potency information. Nineteen chemicals covering a broad spectrum of genotoxic modes of action were tested in an in vitro MN test using TK6 cells using the same study protocol. Several of these chemicals were considered to need metabolic activation, and these were administered in the presence of S9. The Benchmark dose (BMD) approach was applied using the dose-response modeling program PROAST to estimate the genotoxic potency from the in vitro data. The resulting in vitro BMDs were compared with previously derived BMDs from in vivo MN and carcinogenicity studies. A proportional correlation was observed between the BMDs from the in vitro MN and the BMDs from the in vivo MN assays. Further, a clear correlation was found between the BMDs from in vitro MN and the associated BMDs for malignant tumors. Although these results are based on only 19 compounds, they show that genotoxicity potencies estimated from in vitro tests may result in useful information regarding in vivo genotoxic potency, as well as expected cancer potency. Extension of the number of compounds and further investigation of metabolic activation (S9) and of other toxicokinetic factors would be needed to validate our initial conclusions. However, this initial work suggests that this approach could be used for in vitro to in vivo extrapolations which would support the reduction of animals used in research (3Rs: replacement, reduction, and refinement). © The Author 2015. Published by Oxford University Press on behalf of the Society of Toxicology.

  10. In vitro developmental toxicity test detects inhibition of stem cell differentiation by silica nanoparticles.

    NARCIS (Netherlands)

    Park, M.V.; Annema, W.; Salvati, A.; Lesniak, A.; Elsaesser, A.; Barnes, C.; McKerr, G.; Howard, C.; Lynch, I.; Dawson, K.; Piersma, A.H.; de Jong, W.H.

    2009-01-01

    While research into the potential toxic properties of nanomaterials is now increasing, the area of developmental toxicity has remained relatively uninvestigated. The embryonic stem cell test is an in vitro screening assay used to investigate the embryotoxic potential of chemicals by determining

  11. A rapid in vitro screening system for the identification and evaluation of anticancer drugs

    International Nuclear Information System (INIS)

    Kao, J.W.; Collins, J.L.

    1989-01-01

    We report the development of an in vitro screening system that can be used to identify new anticancer drugs that are specifically cytotoxic for dividing cells. The screening system takes advantage of the potential of many cell lines, including tumor cells, to stop dividing when they are plated at high cell density. The cytotoxic effects of anticancer drugs on dividing (i.e., cells plated at low cell density) and nondividing cells (i.e., cells plated at high cell density) is measured by the incorporation of 51Cr. This in vitro system was evaluated by measuring the cytotoxic effects of the anticancer drugs cisplatin, thiotepa, doxorubicin, methotrexate, and vinblastine on the cell lines B/C-N, ME-180, and MCF-7. In this in vitro system the concentrations of the anticancer drugs that produced significant cytotoxicity on only dividing cells are similar to the concentrations that are used clinically. The fact that this in vitro system is rapid, simple, applicable to many cell types, and able to predict effective concentrations of anticancer drugs should make it useful for the screening of new anticancer drugs and for the design of preclinical studies

  12. The limits of testing particle-mediated oxidative stress in vitro in predicting diverse pathologies; relevance for testing of nanoparticles

    Directory of Open Access Journals (Sweden)

    Gulumian Mary

    2009-04-01

    Full Text Available Abstract In vitro studies with particles are a major staple of particle toxicology, generally used to investigate mechanisms and better understand the molecular events underlying cellular effects. However, there is ethical and financial pressure in nanotoxicology, the new sub-specialty of particle toxicology, to avoid using animals. Therefore an increasing amount of studies are being published using in vitro approaches and such studies require careful interpretation. We point out here that 3 different conventional pathogenic particle types, PM10, asbestos and quartz, which cause diverse pathological effects, have been reported to cause very similar oxidative stress effects in cells in culture. We discuss the likely explanation and implications of this apparent paradox, and its relevance for testing in nanotoxicology.

  13. Potency testing of veterinary vaccines: the way from in vivo to in vitro.

    Science.gov (United States)

    Romberg, Judith; Lang, Stefan; Balks, Elisabeth; Kamphuis, Elisabeth; Duchow, Karin; Loos, Daniela; Rau, Henriette; Motitschke, Andreas; Jungbäck, Carmen

    2012-01-01

    Current quality control of inactivated animal vaccines still focuses on the potency of final products in a batch-wise manner. Animal welfare concerns as well as scientific considerations have led to the '3Rs-concept' that comprises the refinement of animal procedures, the reduction of animal numbers, and the replacement of animal models. Although the 3Rs-concept has been widely accepted as a fundamental principle, the number of approved alternatives for in vivo tests is still limited. To promote further progress, the international scientific workshop 'Potency Testing of Veterinary Vaccines: The Way from in vivo to in vitro' was held at the Paul-Ehrlich-Institut in Langen, Germany, on 01-03 December 2010. More than 130 participants from industry, academia and regulatory authorities discussed the current state of the 3Rs-concept, examples of its successful implementation as well as still existing hurdles. Special emphasis was laid on the 'consistency approach' that aims to ensure relevant quality attributes of vaccine batches by in vitro analyses during production rather than by in vivo potency tests on the final product. This report provides an overview of the insights gained, including the recommendations produced at the end of the workshop. Copyright © 2011. Published by Elsevier Ltd.. All rights reserved.

  14. Body-on-a-chip systems for animal-free toxicity testing.

    Science.gov (United States)

    Mahler, Gretchen J; Esch, Mandy B; Stokol, Tracy; Hickman, James J; Shuler, Michael L

    2016-10-01

    Body-on-a-chip systems replicate the size relationships of organs, blood distribution and blood flow, in accordance with human physiology. When operated with tissues derived from human cell sources, these systems are capable of simulating human metabolism, including the conversion of a prodrug to its effective metabolite, as well as its subsequent therapeutic actions and toxic side-effects. The system also permits the measurement of human tissue electrical and mechanical reactions, which provide a measure of functional response. Since these devices can be operated with human tissue samples or with in vitro tissues derived from induced pluripotent stem cells (iPS), they can play a significant role in determining the success of new pharmaceuticals, without resorting to the use of animals. By providing a platform for testing in the context of human metabolism, as opposed to animal models, the systems have the potential to eliminate the use of animals in preclinical trials. This article will review progress made and work achieved as a direct result of the 2015 Lush Science Prize in support of animal-free testing. 2016 FRAME.

  15. An in vitro drug sensitivity test using a higher 3H-TdR incorporation and a modified human tumor stem cell assay

    International Nuclear Information System (INIS)

    Wang Enzhong

    1991-01-01

    An in vitro drug sensitivity test was developed to evaluate the lethal effects of drugs on human pulmonary carcinoma cells (HPCC). This method was a variant and combination of Human Tumor Stem (HTSCA) and a short-term test using 3 H-TdR incorporation. It consisted of a cell containing liquid top layer and a soft agar bottom layer in 24-well microplates. The medium was RPMI 1640 supplemented with 20% malignant pleural effusion, which could enhance 3 H-TdR incorporation into malignant cells. When 50%, 40%, 30% and 30% of cell survival rate defined as sensitivity-threshold for VCR, MMC, DDP and ADM respectively, in the vitro effectiveness were close to those of clinical single-drug treatment in HPCC by Wright et al. This method was also compared with HTSCA in ten human lung cancer cell lines and four pulmonary carcinoma tissues. The agreement rates were 83% and 100% respectively. Thus we presume this system is more useful for oncological clinics than the others

  16. In vitro culture of pre-implanted mouse embryos. A model system for studying combined effects

    International Nuclear Information System (INIS)

    Streffer, C.; Beuningen, D. van; Molls, M.; Pon, A.; Schulz, S.; Zamboglou, N.

    1978-01-01

    Studies on combined effects, e.g. interaction between chemical toxicants and ionizing radiation, are difficult to perform, as they are dependent on many factors (substance concentration, radiation dose, sequence of treatments, etc.). In order to obtain data from such studies it is necessary to establish a comparatively simple experimental model system. We have established such a model system by studying combined effects on pre-implanted mouse embryos cultured in vitro. This system has the following advantages: (1) The embryos can be cultivated for several days in vitro; (2) Their physiological intactness can be tested; and (3) Cell proliferation, cell killing and chromosomal damage can be investigated comparatively easily. The embryos are isolated at the 2-cell stage and incubated in a culture medium in vitro. The development of the embryos is followed under the microscope until the development of blastocysts or the hatching of blastocysts is observed. These blastocysts can be transplanted to fostered mice and the development of normal animals determined. The proliferation kinetics can be studied easily, and the methods are described. A method has also been developed to measure the DNA content of individual cells by microscope fluorometry. After treatment of the embryos with ionizing radiation or drugs the release of micronuclei has been observed from the cell nuclei, which is an expression for chromosomal damage. Substances or radionuclides can be added to the culture medium or external irradiation can be performed during the culture period. Also the combined effects of radiation and heating can be studied. The effects of X-rays and tritiated compounds have also been investigated. The combined effects of radiation with antibiotics such as actinomycin D, and environmental toxicants such as lead, have been determined. The system described has been useful to evaluate cytological, teratogenic and cytogenetic effects

  17. Microleakage of composite crowns luted on CAD/CAM-milled human molars: a new method for standardized in vitro tests.

    Science.gov (United States)

    Schlenz, Maximiliane Amelie; Schmidt, Alexander; Rehmann, Peter; Niem, Thomas; Wöstmann, Bernd

    2018-04-24

    To investigate debonding of full crowns made of CAD/CAM composites, CAD/CAM technology was applied to manufacture standardized test abutments to increase the reproducibility of human teeth used in in vitro studies. A virtual test abutment and the corresponding virtual crown were designed and two STL data sets were generated. Sixty-four human third molars and CAD/CAM blocks were milled using a CNC machine. Crowns of four different composite blocks (Lava Ultimate (LU), Brilliant Crios (BC), Cerasmart (CS), Experimental (EX)) were adhesively bonded with their corresponding luting system (LU: Scotchbond Universal/RelyX Ultimate; BC: One Coat 7 Universal/DuoCem; CS: G-PremioBond/G-Cem LinkForce; EX: Experimental-Bond/Experimental-Luting-Cement). Half of the specimens were chemical-cured (CC) and the others were light-cured (LC). Afterwards, specimens were artificially aged in a chewing simulator (WL-tec, 1 million cycles, 50-500 N, 2 Hz, 37 °C). Finally, a dye penetration test was used to detect debonding. For inspection, the specimens were sliced, and penetration depth was measured with a digital microscope. Data were analyzed with the Mann-Whitney U test. No cases of total debonding were observed after cyclic loading. However, the LC specimens showed a significantly lower amount of leakage than the CC ones (p CAD/CAM technology in highly standardized test abutments for in vitro testing. For CAD/CAM composites, light curing should be performed. The success of a restoration depends on the long-term sealing ability of the luting materials, which avoids debonding along with microleakage. For CAD/CAM composites, separate light curing of the adhesive and luting composite is highly recommended.

  18. In vitro testing of curcumin based composites coatings as antitumoral systems against osteosarcoma cells

    Science.gov (United States)

    Tirca, I.; Mitran, V.; Marascu, V.; Brajnicov, S.; Ion, V.; Stokker-Cheregi, F.; Popovici, I. A.; Cimpean, A.; Dinca, V.; Dinescu, M.

    2017-12-01

    In this work, we propose a new design for biodegradable composite coatings obtained by laser methods, which are aimed at evaluating the effects of active antitumoral elements on osteosarcoma cells. Our approach relies on embedding curcumin, which is a natural polyphenol having antitumoral properties, within biodegradable copolymer coatings (i.e. polyvinyl alcohol-polyethylene glycol - PVA-PEG) by using matrix assisted pulsed laser evaporation (MAPLE). The structural and morphological characteristics of the coatings were tailored by using different solvents (water, ethanol, benzene, dimethylsufoxide) as deposition matrix. The morphological characteristics of the resulting films were investigated by atomic force microscopy (AFM), whereas their chemical composition was characterized by Fourier transform infrared spectroscopy (FTIR). These characteristics were correlated with the degradation behavior by using ellipsometry (SE) and AFM measurements data. The in vitro study of the MG-63 osteosarcoma cell behavior indicates that the developed hybrid coatings significantly decreased osteosarcoma cell viability and proliferation potential. The physico-chemical characteristics of the thin films, along with the preliminary in vitro analyses, suggest that our developed polymeric hybrid coatings represent an efficient way to tackle the design of antitumoral surfaces, with applications in biomedicine.

  19. Immunochemical faecal occult blood tests have superior stability and analytical performance characteristics over guaiac-based tests in a controlled in vitro study.

    LENUS (Irish Health Repository)

    Lee, Chun Seng

    2011-06-01

    The aims of this study were (1) to determine the measurement accuracy of a widely used guaiac faecal occult blood test (gFOBT) compared with an immunochemical faecal occult blood test (iFOBT) during in vitro studies, including their analytical stability over time at ambient temperature and at 4°C; and (2) to compare analytical imprecision and other characteristics between two commercially available iFOBT methods.

  20. Synthesis of milligram quantities of proteins using a reconstituted in vitro protein synthesis system.

    Science.gov (United States)

    Kazuta, Yasuaki; Matsuura, Tomoaki; Ichihashi, Norikazu; Yomo, Tetsuya

    2014-11-01

    In this study, the amount of protein synthesized using an in vitro protein synthesis system composed of only highly purified components (the PURE system) was optimized. By varying the concentrations of each system component, we determined the component concentrations that result in the synthesis of 0.38 mg/mL green fluorescent protein (GFP) in batch mode and 3.8 mg/mL GFP in dialysis mode. In dialysis mode, protein concentrations of 4.3 and 4.4 mg/mL were synthesized for dihydrofolate reductase and β-galactosidase, respectively. Using the optimized system, the synthesized protein represented 30% (w/w) of the total protein, which is comparable to the level of overexpressed protein in Escherichia coli cells. This optimized reconstituted in vitro protein synthesis system may potentially be useful for various applications, including in vitro directed evolution of proteins, artificial cell assembly, and protein structural studies. Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  1. Statistical comparative study on a combined radioiodine test and extended protirelin test and correlation with the common in vitro parameters of hyroid function

    International Nuclear Information System (INIS)

    Kraemer, H.A.

    1982-01-01

    Using the data of 339 patients, the following parameters of thyroid function were statistically evaluated. The in vitro parameters ET 3 U, TT 4 (D), FT 4 -index and PB 127 I and the radioiodine test with determination of PB 131 I before i.v. injection of 400 μg protirelin (DHP) and 120 minutes after the injection. There was no correlation between the percentage Change of the PB 121 I level 120 min after protirelin (DHP) administration and the percentage change of the TSH level 30 min after protirelin (DTP1) administration. The accuracies of the in vitro parameters ET 3 U, TT 4 (D) and FT 4 -index on the one hand and the extended protirelin test on the other hand were compared. (orig./MG) [de

  2. Light-Emitting Diode-Based Illumination System for In Vitro Photodynamic Therapy

    Directory of Open Access Journals (Sweden)

    Defu Chen

    2012-01-01

    Full Text Available The aim of this study is to develop a light-emitting diode- (LED- based illumination system that can be used as an alternative light source for in vitro photodynamic therapy (PDT. This illumination system includes a red LED array composed of 70 LEDs centered at 643 nm, an air-cooling unit, and a specific-designed case. The irradiance as a function of the irradiation distance between the LED array and the sample, the homogeneity and stability of irradiation, and the effect of long-time irradiation on culture medium temperature were characterized. Furthermore, the survival rate of the CNE1 cells that sensitized with 5-aminolevulinic acid after PDT treatment was evaluated to demonstrate the efficiency of the new LED-based illumination system. The obtained results show that the LED-based illumination system is a promising light source for in vitro PDT that performed in standard multiwell plate.

  3. In vitro fatigue tests and in silico finite element analysis of dental implants with different fixture/abutment joint types using computer-aided design models.

    Science.gov (United States)

    Yamaguchi, Satoshi; Yamanishi, Yasufumi; Machado, Lucas S; Matsumoto, Shuji; Tovar, Nick; Coelho, Paulo G; Thompson, Van P; Imazato, Satoshi

    2018-01-01

    The aim of this study was to evaluate fatigue resistance of dental fixtures with two different fixture-abutment connections by in vitro fatigue testing and in silico three-dimensional finite element analysis (3D FEA) using original computer-aided design (CAD) models. Dental implant fixtures with external connection (EX) or internal connection (IN) abutments were fabricated from original CAD models using grade IV titanium and step-stress accelerated life testing was performed. Fatigue cycles and loads were assessed by Weibull analysis, and fatigue cracking was observed by micro-computed tomography and a stereomicroscope with high dynamic range software. Using the same CAD models, displacement vectors of implant components were also analyzed by 3D FEA. Angles of the fractured line occurring at fixture platforms in vitro and of displacement vectors corresponding to the fractured line in silico were compared by two-way ANOVA. Fatigue testing showed significantly greater reliability for IN than EX (pimplant fixture platforms. FEA demonstrated that crack lines of both implant systems in vitro were observed in the same direction as displacement vectors of the implant fixtures in silico. In silico displacement vectors in the implant fixture are insightful for geometric development of dental implants to reduce complex interactions leading to fatigue failure. Copyright © 2017 Japan Prosthodontic Society. Published by Elsevier Ltd. All rights reserved.

  4. Testing for developmental neurotoxicity using a battery of in vitro assays for key cellular events in neurodevelopment.

    Science.gov (United States)

    Harrill, Joshua A; Freudenrich, Theresa; Wallace, Kathleen; Ball, Kenneth; Shafer, Timothy J; Mundy, William R

    2018-04-05

    Medium- to high-throughput in vitro assays that recapitulate the critical processes of nervous system development have been proposed as a means to facilitate rapid testing and identification of chemicals which may affect brain development. In vivo neurodevelopment is a complex progression of distinct cellular processes. Therefore, batteries of in vitro assays that model and quantify effects on a variety of neurodevelopmental processes have the potential to identify chemicals which may affect brain development at different developmental stages. In the present study, the results of concentration-response screening of 67 reference chemicals in a battery of high content imaging and microplate reader-based assays that evaluate neural progenitor cell proliferation, neural proginitor cell apoptosis, neurite initiation/outgrowth, neurite maturation and synaptogenesis are summarized and compared. The assay battery had a high degree of combined sensitivity (87%) for categorizing chemicals known to affect neurodevelopment as active and a moderate degree of combined specificity (71%) for categorizing chemicals not associated with affects on neurodevelopment as inactive. The combined sensitivity of the assay battery was higher compared to any individual assay while the combined specificity of the assay battery was lower compared to any individual assay. When selectivity of effects for a neurodevelopmental endpoint as compared to general cytotoxicity was taken into account, the combined sensitivity of the assay battery decreased (68%) while the combined specificity increased (93%). The identity and potency of chemicals identified as active varied across the assay battery, underscoring the need for use of a combination of diverse in vitro models to comprehensively screen chemicals and identify those which potentially affect neurodevelopment. Overall, these data indicate that a battery of assays which address many different processes in nervous system development may be used to

  5. Cost effectiveness of ovarian reserve testing in in vitro fertilization : a Markov decision-analytic model

    NARCIS (Netherlands)

    Moolenaar, Lobke M.; Broekmans, Frank J. M.; van Disseldorp, Jeroen; Fauser, Bart C. J. M.; Eijkemans, Marinus J. C.; Hompes, Peter G. A.; van der Veen, Fulco; Mol, Ben Willem J.

    2011-01-01

    Objective: To compare the cost effectiveness of ovarian reserve testing in in vitro fertilization (IVF). Design: A Markov decision model based on data from the literature and original patient data. Setting: Decision analytic framework. Patient(s): Computer-simulated cohort of subfertile women aged

  6. Absorbable magnesium-based stent: physiological factors to consider for in vitro degradation assessments.

    Science.gov (United States)

    Wang, Juan; Smith, Christopher E; Sankar, Jagannathan; Yun, Yeoheung; Huang, Nan

    2015-03-01

    Absorbable metals have been widely tested in various in vitro settings using cells to evaluate their possible suitability as an implant material. However, there exists a gap between in vivo and in vitro test results for absorbable materials. A lot of traditional in vitro assessments for permanent materials are no longer applicable to absorbable metallic implants. A key step is to identify and test the relevant microenvironment and parameters in test systems, which should be adapted according to the specific application. New test methods are necessary to reduce the difference between in vivo and in vitro test results and provide more accurate information to better understand absorbable metallic implants. In this investigative review, we strive to summarize the latest test methods for characterizing absorbable magnesium-based stent for bioabsorption/biodegradation behavior in the mimicking vascular environments. Also, this article comprehensively discusses the direction of test standardization for absorbable stents to paint a more accurate picture of the in vivo condition around implants to determine the most important parameters and their dynamic interactions.

  7. In vitro Differentiation of Functional Human Skeletal Myotubes in a Defined System.

    Science.gov (United States)

    Guo, Xiufang; Greene, Keshel; Akanda, Nesar; Smith, Alec; Stancescu, Maria; Lambert, Stephen; Vandenburgh, Herman; Hickman, James

    2014-01-01

    In vitro human skeletal muscle systems are valuable tools for the study of human muscular development, disease and treatment. However, published in vitro human muscle systems have so far only demonstrated limited differentiation capacities. Advanced differentiation features such as cross-striations and contractility have only been observed in co-cultures with motoneurons. Furthermore, it is commonly regarded that cultured human myotubes do not spontaneously contract, and any contraction has been considered to originate from innervation. This study developed a serum-free culture system in which human skeletal myotubes demonstrated advanced differentiation. Characterization by immunocytochemistry, electrophysiology and analysis of contractile function revealed these major features: A) well defined sarcomeric development, as demonstrated by the presence of cross-striations. B) finely developed excitation-contraction coupling apparatus characterized by the close apposition of dihydropyridine receptors on T-tubules and Ryanodine receptors on sarcoplasmic reticulum membranes. C) spontaneous and electrically controlled contractility. This report not only demonstrates an improved level of differentiation of cultured human skeletal myotubes, but also provides the first published evidence that such myotubes are capable of spontaneous contraction. Use of this functional in vitro human skeletal muscle system would advance studies concerning human skeletal muscle development and physiology, as well as muscle-related disease and therapy.

  8. Validation of an in vitro digestive system for studying macronutrient decomposition in humans.

    Science.gov (United States)

    Kopf-Bolanz, Katrin A; Schwander, Flurina; Gijs, Martin; Vergères, Guy; Portmann, Reto; Egger, Lotti

    2012-02-01

    The digestive process transforms nutrients and bioactive compounds contained in food to physiologically active compounds. In vitro digestion systems have proven to be powerful tools for understanding and monitoring the complex transformation processes that take place during digestion. Moreover, the investigation of the physiological effects of certain nutrients demands an in vitro digestive process that is close to human physiology. In this study, human digestion was simulated with a 3-step in vitro process that was validated in depth by choosing pasteurized milk as an example of a complex food matrix. The evolution and decomposition of the macronutrients was followed over the entire digestive process to the level of intestinal enterocyte action, using protein and peptide analysis by SDS-PAGE, reversed-phase HPLC, size exclusion HPLC, and liquid chromatography-MS. The mean peptide size after in vitro digestion of pasteurized milk was 5-6 amino acids (AA). Interestingly, mostly essential AA (93.6%) were released during in vitro milk digestion, a significantly different relative distribution compared to the total essential AA concentration of bovine milk (44.5%). All TG were degraded to FFA and monoacylglycerols. Herein, we present a human in vitro digestion model validated for its ability to degrade the macronutrients of dairy products comparable to physiological ranges. It is suited to be used in combination with a human intestinal cell culture system, allowing ex vivo bioavailability measurements and assessment of the bioactive properties of food components.

  9. Particular Characterisation of an In-Vitro-DTH Test to Monitor Cellular Immunity - Applications for Patient Care and Space Flight

    Science.gov (United States)

    Feurecker, M.; Mayer, W.; Gruber, M.; Muckenthaler, F.; Draenert, R.; Bogner, J.; Kaufmann, I.; Crucian, B.; Rykova, M.; Morukov, B.; hide

    2010-01-01

    Goal:i) Characterization of the role of the main immune reactive cell types contributing to the cellular immune response in the in-vitro DTH and ii) Validation of the in-vitro DTH under different clinical and field conditions. Methods:As positive control whole blood was incubated in the in-vitro DTH, supernatants were gathered after 12, 24 and 48h. Readout parameters of this test are cytokines in the assay's supernatant. To determine the role of T-cells, monocytes and natural killer (NK), these cell populations were depleted using magnetic beads prior to in-vitro-DTH incubation. Validation of the test has occurred under clinical (HIV-patients, ICU) and field-conditions (parabolic/space-flights, confinement). Results:T-cell depletion abandoned almost any IL-2 production and reduced IFN-gamma production irrespective of the type of antigen, whereas CD56 depleted cultures tended to lower IL-2 secretion and IFN-gamma and to parallel a IL-10-increase after viral challenge. This IL-10-increase was seen also in CD14-depleted setups. DTH read-out was significantly different under acute stress (parabolic flight) or chronic stress (ISS), respectively. Preliminary data of HIV infected patients demonstrate that this test can display the contemporary immune status during an antiviral therapy. Conclusion:The in-vitro DTH mirrors adaptive and innate immune activation and may serve as tool also for longitudinal follow up of Th1/Th2 weighed immune response under adverse life conditions on earth and in space. It is planned to implement the assay in the on the ISS (MoCISS).

  10. In vitro evaluation of radio-labeled aerosol delivery via a variable-flow infant CPAP system.

    Science.gov (United States)

    Farney, Kimberly D; Kuehne, Brandon T; Gibson, Laurie A; Nelin, Leif D; Shepherd, Edward G

    2014-03-01

    Nasal CPAP is widely used in neonatal ICUs. Aerosolized medications such as inhaled steroids and β agonists are commonly administered in-line through nasal CPAP, especially to infants with bronchopulmonary dysplasia. We hypothesized that aerosol delivery to the lungs via variable-flow nasal CPAP in an in vitro model would be unreliable, and that the delivery would depend on the position of the aerosol generator within the nasal CPAP circuit. We used a system that employed a test lung placed in a plastic jar and subjected to negative pressure. Simulated inspiration effort was measured with a heated-wire anemometer. We used technetium-99m-labeled diethylene triamine penta-acetic acid as our aerosol. The nebulizer was placed either close to the humidifier or close to the nasal prongs in the circuit, and patient effort was simulated with a minute ventilation of 0.4 L/min. Relative aerosol delivery to the infant test lung with the nebulizer close to the humidifier was extremely low (0.3 ± 0.4%), whereas placing the nebulizer close to the nasal prongs resulted in significantly (P CPAP was negligible in this in vitro setup; however, such delivery was significantly improved by locating the aerosol generator closer to the nasal CPAP interface.

  11. Gas transport during in vitro and in vivo preclinical testing of inert gas therapies

    Directory of Open Access Journals (Sweden)

    Ira Katz

    2016-01-01

    Full Text Available New gas therapies using inert gases such as xenon and argon are being studied, which require in vitro and in vivo preclinical experiments. Examples of the kinetics of gas transport during such experiments are analyzed in this paper. Using analytical and numerical models, we analyze an in vitro experiment for gas transport to a 96 cell well plate and an in vivo delivery to a small animal chamber, where the key processes considered are the wash-in of test gas into an apparatus dead volume, the diffusion of test gas through the liquid media in a well of a cell test plate, and the pharmacokinetics in a rat. In the case of small animals in a chamber, the key variable controlling the kinetics is the chamber wash-in time constant that is a function of the chamber volume and the gas flow rate. For cells covered by a liquid media the diffusion of gas through the liquid media is the dominant mechanism, such that liquid depth and the gas diffusion constant are the key parameters. The key message from these analyses is that the transport of gas during preclinical experiments can be important in determining the true dose as experienced at the site of action in an animal or to a cell.

  12. Triple co-culture cell model as an in vitro model for oral particulate vaccine systems

    DEFF Research Database (Denmark)

    Nielsen, Line Hagner; De Rossi, C.; Lehr, C-M.

    ; this was not observed with ovalbumin and blank solution. An example of the results is shown in Figure 2 for IL-17A. An established co-culture of Caco-2, THP-1 and MUTZ-3 cells showed promise as an in vitro model for testing of oral vaccine formulations. Mobility of co-culture immune cells as well as cytokine production......A triple co-culture cell model of Caco-2 cells, dendritic cells and macrophages (Figure 1) has previously been developed for studying intestinal permeability in a state of inflammation [1],[2]. The aim of this study was to investigate the applicability of this cell model for testing...... the model antigen ovalbumin was spray dried to obtain a particulate vaccine model system for testing in the cell model. The precursors were shown to form cubosomes when dispersed in aqueous medium, and was therefore used as the vaccine formulation for testing on the co-cultures. After 11 days, the TEER...

  13. Comparison of in vitro test systems using bacterial and mammalian cells for genotoxicity assessment within the "health-related indication value (HRIV) concept.

    Science.gov (United States)

    Prantl, Eva-Maria; Kramer, Meike; Schmidt, Carsten K; Knauer, Martina; Gartiser, Stefan; Shuliakevich, Aliaksandra; Milas, Julia; Glatt, Hansruedi; Meinl, Walter; Hollert, Henner

    2018-02-01

    In numerous cases, the German health-related indication value (HRIV) concept has proved its practicability for the assessment of drinking water relevant trace substances (Umweltbundesamt 2003). The HRIV is based on the toxicological profile of a substance. An open point of the HRIV concept has been the assignment of standardized test procedures to be used for the assessment. The level of the HRIV is at its lowest as soon as the genotoxicity of the substance is detected. As a single test on its own, it is not sufficient enough to assess the human toxicological relevance of a genotoxic effect or exclude it in the case of a negative result; a reasonable test battery was required, technically oriented towards the already harmonized international, hierarchical evaluation for toxicological assessment of chemicals. Therefore, an important aim of this project was to define a strategy for the genotoxicological assessment of anthropogenic trace substances. The basic test battery for genotoxicity of micropollutants in drinking water needs to fulfill several requirements. Although quick test results are needed for the determination of HRIV, a high degree of transferability to human genotoxicity should be ensured. Therefore, an in vitro genotoxicity test battery consisting of the Ames fluctuation test with two tester strains (ISO 11350), the umu test and the micronucleus test, or from the Ames test with five tester strains (OECD 471) and the micronucleus test is proposed. On the basis of selected test substances, it could be shown that the test battery leads to positive, indifferent, and negative results. Given indifferent results, the health authority and the water supplier must assume that it is a genotoxic substance. Genetically modified tester strains are being sensitive to different chemical classes by expression of selected mammalian key enzymes for example nitroreductase, acetyltransferase, and glutathione-S-transferase. These strains may provide valuable additional

  14. Good cell culture practices &in vitro toxicology.

    Science.gov (United States)

    Eskes, Chantra; Boström, Ann-Charlotte; Bowe, Gerhard; Coecke, Sandra; Hartung, Thomas; Hendriks, Giel; Pamies, David; Piton, Alain; Rovida, Costanza

    2017-12-01

    Good Cell Culture Practices (GCCP) is of high relevance to in vitro toxicology. The European Society of Toxicology In Vitro (ESTIV), the Center for Alternatives for Animal Testing (CAAT) and the In Vitro Toxicology Industrial Platform (IVTIP) joined forces to address by means of an ESTIV 2016 pre-congress session the different aspects and applications of GCCP. The covered aspects comprised the current status of the OECD guidance document on Good In Vitro Method Practices, the importance of quality assurance for new technological advances in in vitro toxicology including stem cells, and the optimized implementation of Good Manufacturing Practices and Good Laboratory Practices for regulatory testing purposes. General discussions raised the duality related to the difficulties in implementing GCCP in an academic innovative research framework on one hand, and on the other hand, the need for such GCCP principles in order to ensure reproducibility and robustness of in vitro test methods for toxicity testing. Indeed, if good cell culture principles are critical to take into consideration for all uses of in vitro test methods for toxicity testing, the level of application of such principles may depend on the stage of development of the test method as well as on the applications of the test methods, i.e., academic innovative research vs. regulatory standardized test method. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. Cost effectiveness of ovarian reserve testing in in vitro fertilization: a Markov decision-analytic model

    NARCIS (Netherlands)

    Moolenaar, Lobke M.; Broekmans, Frank J. M.; van Disseldorp, Jeroen; Fauser, Bart C. J. M.; Eijkemans, Marinus J. C.; Hompes, Peter G. A.; van der Veen, Fulco; Mol, Ben Willem J.

    2011-01-01

    To compare the cost effectiveness of ovarian reserve testing in in vitro fertilization (IVF). A Markov decision model based on data from the literature and original patient data. Decision analytic framework. Computer-simulated cohort of subfertile women aged 20 to 45 years who are eligible for IVF.

  16. Summary, the 16th quality control survey for radioisotope in vitro tests in Japan, 1994

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1995-11-01

    The results of the 16th quality control survey for radioisotope in vitro tests in Japan (1994) are summarized. Of 399 medical facilities conducting radioisotope in vitro tests, 201 were enrolled in this study. Forty items including ACTH and {alpha}-fetoprotein were selected as the subjects. Freeze-drying samples were sent to the facilities. The quality of assay tubes, duration between fusion of the samples and assay, and the condition of preservation were examined, and those influence on the assay values were studied. Radioimmunoassay, immunoradiometric assay, and other procedures using enzymes, fluorescence, and chemiluminescense were conducted. The assay values of some of the items were significantly influenced by repeated freezing and fusion of the samples. Data were collected from individual items and kits used, and analyzed. The significant difference of values between different facilities and kits used were considered due to difference of assay principle, antibodies used, and standard items. The concentration of the samples needs to be improved. (S.Y.).

  17. In vitro culture systems and acclimatization of Aechmea setigera Mart. ex Schult. & Schult. f. (Bromeliaceae

    Directory of Open Access Journals (Sweden)

    Janaína Medeiros Vasconcelos

    2015-12-01

    Full Text Available Aechmea setigera is an endemic bromeliad from Amazon with ornamental potential. Bromeliads have been propagated by tissue culture. The consistency of the culture medium in vitro multiplication influences the rate of propagation. In this sense, the objective of this study was to evaluate different culture systems with the use of 6-benzylaminopurine (BAP on in vitro propagation and the effect of different substrates in acclimatization of plantlets Aechmea setigera. In vitro germinated seedlings were inoculated in MS medium in liquid stationary, semisolid, double-phase systems, plus 6-benzylaminopurine (BAP in different concentrations (0, 2.2, 4.4, 8.8 and 17.7 μM. The ex vitro rooting and acclimatization were performed on substrate Plantmax Forest ®, vermiculite and sawdust eucalyptus. After three successive subcultures, the double-phase system showed a higher number of regenerated shoots in comparison to other systems. Acclimatization using the combination of commercial substrate Plantmax Forest ® and vermiculite favored the growth of micropropagated plants. The use of a culture medium double-phase without growth regulator, and the rooting in acclimatization are feasible strategy for the micropropagation of A. setigera. Indexação

  18. Trends in the development of microfluidic cell biochips for in vitro hepatotoxicity.

    Science.gov (United States)

    Baudoin, Régis; Corlu, Anne; Griscom, Laurent; Legallais, Cécile; Leclerc, Eric

    2007-06-01

    Current developments in the technological fields of liver tissue engineering, bioengineering, biomechanics, microfabrication and microfluidics have lead to highly complex and pertinent new tools called "cell biochips" for in vitro toxicology. The purpose of "cell biochips" is to mimic organ tissues in vitro in order to partially reduce the amount of in vivo testing. These "cell biochips" consist of microchambers containing engineered tissue and living cell cultures interconnected by a microfluidic network, which allows the control of microfluidic flows for dynamic cultures, by continuous feeding of nutrients to cultured cells and waste removal. Cell biochips also allow the control of physiological contact times of diluted molecules with the tissues and cells, for rapid testing of sample preparations or specific addressing. Cell biochips can be situated between in vitro and in vivo testing. These types of systems can enhance functionality of cells by mimicking the tissue architecture complexities when compared to in vitro analysis but at the same time present a more rapid and simple process when compared to in vivo testing procedures. In this paper, we first introduce the concepts of microfluidic and biochip systems based on recent progress in microfabrication techniques used to mimic liver tissue in vitro. This includes progress and understanding in biomaterials science (cell culture substrate), biomechanics (dynamic cultures conditions) and biology (tissue engineering). The development of new "cell biochips" for chronic toxicology analysis of engineered tissues can be achieved through the combination of these research domains. Combining these advanced research domains, we then present "cell biochips" that allow liver chronic toxicity analysis in vitro on engineered tissues. An extension of the "cell biochip" idea has also allowed "organ interactions on chip", which can be considered as a first step towards the replacement of animal testing using a combined liver

  19. Small test SDHW systems

    DEFF Research Database (Denmark)

    Vejen, Niels Kristian

    1999-01-01

    Three small test SDHW systems was tested in a laboratory test facility.The three SDHW systems where all based on the low flow principe and a mantle tank but the design of the systems where different.......Three small test SDHW systems was tested in a laboratory test facility.The three SDHW systems where all based on the low flow principe and a mantle tank but the design of the systems where different....

  20. A Novel In Vitro System for Comparative Analyses of Bone Cells and Bacteria under Electrical Stimulation

    Directory of Open Access Journals (Sweden)

    Thomas Josef Dauben

    2016-01-01

    Full Text Available Electrical stimulation is a promising approach to enhance bone regeneration while having potential to inhibit bacterial growth. To investigate effects of alternating electric field stimulation on both human osteoblasts and bacteria, a novel in vitro system was designed. Electric field distribution was simulated numerically and proved by experimental validation. Cells were stimulated on Ti6Al4V electrodes and in short distance to electrodes. Bacterial growth was enumerated in supernatant and on the electrode surface and biofilm formation was quantified. Electrical stimulation modulated gene expression of osteoblastic differentiation markers in a voltage-dependent manner, resulting in significantly enhanced osteocalcin mRNA synthesis rate on electrodes after stimulation with 1.4VRMS. While collagen type I synthesis increased when stimulated with 0.2VRMS, it decreased after stimulation with 1.4VRMS. Only slight and infrequent influence on bacterial growth was observed following stimulations with 0.2VRMS and 1.4VRMS after 48 and 72 h, respectively. In summary this novel test system is applicable for extended in vitro studies concerning definition of appropriate stimulation parameters for bone cell growth and differentiation, bacterial growth suppression, and investigation of general effects of electrical stimulation.

  1. In vitro systems: standardization of endpoints

    International Nuclear Information System (INIS)

    Dewey, W.C.

    1979-01-01

    Principles are discussed for utilizing cell culture systems to assay for interactions between drugs and radiation. Emphasis is based on the necessity of using synchronous cultures to determine whether the interaction is independent, additive, or synergistic, and to determine the effect of drugs on recovery from sublethal x-ray damage (SLD). Furthermore, in studies of drug effects on SLD, adequate controls must be included to distinguish between effects associated with the interaction of 2 x-ray doses and the effects associated with interaction of a second x-ray dose with prior drug exposure. Finally, different methods of expressing drug exposure are reviewed, and associated problems related to predicting in vivo effects from in vitro results are mentioned

  2. Thyroid endocrine system disruption by pentachlorophenol: an in vitro and in vivo assay.

    Science.gov (United States)

    Guo, Yongyong; Zhou, Bingsheng

    2013-10-15

    The present study aimed to evaluate the disruption caused to the thyroid endocrine system by pentachlorophenol (PCP) using in vitro and in vivo assays. In the in vitro assay, rat pituitary GH3 cells were exposed to 0, 0.1, 0.3, and 1.0 μM PCP. PCP exposure significantly downregulated basal and triiodothyronine (T3)-induced Dio 1 transcription, indicating the antagonistic activity of PCP in vitro. In the in vivo assay, zebrafish embryos were exposed to 0, 1, 3, and 10 μg/L of PCP until 14 days post-fertilization. PCP exposure resulted in decreased thyroxine (T4) levels, but elevated contents of whole-body T3. PCP exposure significantly upregulated the mRNA expression of genes along hypothalamic-pituitary-thyroid (HPT) axis, including those encoding thyroid-stimulating hormone, sodium/iodide symporter, thyroglobulin, Dio 1 and Dio 2, alpha and beta thyroid hormone receptor, and uridinediphosphate-glucuronosyl-transferase. PCP exposure did not influence the transcription of the transthyretin (TTR) gene. The results indicate that PCP potentially disrupts the thyroid endocrine system both in vitro and in vivo. Copyright © 2013 Elsevier B.V. All rights reserved.

  3. In vitro biocompatibility tests of two commercial types of mineral trioxide aggregate Testes de biocompatibilidade in vitro de duas formas comerciais do agregado de trióxido mineral

    Directory of Open Access Journals (Sweden)

    Daniel Araki Ribeiro

    2005-09-01

    Full Text Available Recently, regular and white mineral trioxide aggregate (MTA are being used in Dentistry as retrofilling materials. Genotoxicity and cytotoxicity tests form an important part of cancer research and risk assessment of potential carcinogens. Thus, the goal of this study was to examine the genotoxicity and cytotoxicity of regular and white MTA in vitro by the single cell gel (comet assay and trypan blue exclusion test, respectively. Mouse lymphoma cells were exposed to two presentation forms of MTA at final concentrations ranging from 1 to 1,000 µg/mL for 3 h at 37°C. The results showed that both compounds tested did not produce genotoxic effects at all concentrations evaluated. Likewise, no statistically significant differences (p > 0.05 were observed in cytotoxicity. Taken together, our results suggest that regular and white MTA are not genotoxins and are not able to interfere in cellular viability as assessed by single cell gel (comet assay and trypan blue assay, respectively.Recentemente, o agregado de trióxido mineral (MTA regular e branco estão sendo utilizados na Odontologia como materiais para obturação retrógrada de canais radiculares. Testes de genotoxicidade e citotoxicidade formam uma importante parte da pesquisa do câncer e da avaliação de risco de carcinógenos potenciais. Assim, o objetivo deste estudo foi examinar a genotoxicidade e citotoxicidade do MTA branco e regular in vitro pelo teste do cometa e teste de exclusão pelo azul de tripan, respectivamente. Células do linfoma murino foram expostas às duas formas de apresentação do MTA nas concentrações finais de 1 a 1.000 µg/mL por 3 horas a 37°C. Os resultados mostraram que ambos os compostos testados não produziram efeito genotóxico em todas as concentrações testadas. Da mesma forma, nenhuma diferença estatisticamente significativa (p > 0,05 foi observada na citotoxicidade. Em suma, nossos resultados sugerem que o MTA regular e branco não são genotoxinas e

  4. Evaluation of a gas in vitro system for predicting methane production in vivo

    DEFF Research Database (Denmark)

    Danielsson, Rebecca; Ramin, Mohammad; Bertilsson, Jan

    2017-01-01

    of samples can be incubated and analyzed at the same time. This study evaluated a recently developed in vitro method for prediction of in vivo CH4 production by examining the relationship between predicted and observed CH4 production values. A total of 49 different diets (observations), used in previous 13......Methane production from ruminant livestock varies with the diet as a result of factors such as dry matter intake, diet composition, and digestibility. To estimate the effect of dietary composition and feed additives, CH4 production can be measured in vitro as a first step because large numbers...... in vivo studies, were selected to include diets varying in nutrient composition. Methane production was measured in all in vivo studies by respiration chambers or the GreenFeed system (C-Lock Inc., Rapid City, SD). Overall, the in vitro system predicted CH4 production well (R2 = 0.96), but the values...

  5. A genomic biomarker signature can predict skin sensitizers using a cell-based in vitro alternative to animal tests

    Directory of Open Access Journals (Sweden)

    Albrekt Ann-Sofie

    2011-08-01

    Full Text Available Abstract Background Allergic contact dermatitis is an inflammatory skin disease that affects a significant proportion of the population. This disease is caused by an adverse immune response towards chemical haptens, and leads to a substantial economic burden for society. Current test of sensitizing chemicals rely on animal experimentation. New legislations on the registration and use of chemicals within pharmaceutical and cosmetic industries have stimulated significant research efforts to develop alternative, human cell-based assays for the prediction of sensitization. The aim is to replace animal experiments with in vitro tests displaying a higher predictive power. Results We have developed a novel cell-based assay for the prediction of sensitizing chemicals. By analyzing the transcriptome of the human cell line MUTZ-3 after 24 h stimulation, using 20 different sensitizing chemicals, 20 non-sensitizing chemicals and vehicle controls, we have identified a biomarker signature of 200 genes with potent discriminatory ability. Using a Support Vector Machine for supervised classification, the prediction performance of the assay revealed an area under the ROC curve of 0.98. In addition, categorizing the chemicals according to the LLNA assay, this gene signature could also predict sensitizing potency. The identified markers are involved in biological pathways with immunological relevant functions, which can shed light on the process of human sensitization. Conclusions A gene signature predicting sensitization, using a human cell line in vitro, has been identified. This simple and robust cell-based assay has the potential to completely replace or drastically reduce the utilization of test systems based on experimental animals. Being based on human biology, the assay is proposed to be more accurate for predicting sensitization in humans, than the traditional animal-based tests.

  6. A genomic biomarker signature can predict skin sensitizers using a cell-based in vitro alternative to animal tests

    Science.gov (United States)

    2011-01-01

    Background Allergic contact dermatitis is an inflammatory skin disease that affects a significant proportion of the population. This disease is caused by an adverse immune response towards chemical haptens, and leads to a substantial economic burden for society. Current test of sensitizing chemicals rely on animal experimentation. New legislations on the registration and use of chemicals within pharmaceutical and cosmetic industries have stimulated significant research efforts to develop alternative, human cell-based assays for the prediction of sensitization. The aim is to replace animal experiments with in vitro tests displaying a higher predictive power. Results We have developed a novel cell-based assay for the prediction of sensitizing chemicals. By analyzing the transcriptome of the human cell line MUTZ-3 after 24 h stimulation, using 20 different sensitizing chemicals, 20 non-sensitizing chemicals and vehicle controls, we have identified a biomarker signature of 200 genes with potent discriminatory ability. Using a Support Vector Machine for supervised classification, the prediction performance of the assay revealed an area under the ROC curve of 0.98. In addition, categorizing the chemicals according to the LLNA assay, this gene signature could also predict sensitizing potency. The identified markers are involved in biological pathways with immunological relevant functions, which can shed light on the process of human sensitization. Conclusions A gene signature predicting sensitization, using a human cell line in vitro, has been identified. This simple and robust cell-based assay has the potential to completely replace or drastically reduce the utilization of test systems based on experimental animals. Being based on human biology, the assay is proposed to be more accurate for predicting sensitization in humans, than the traditional animal-based tests. PMID:21824406

  7. Etoposide; colchicine; mitomycin C and cyclophosphamide tested in the in vitro mammalian cell micronucleus test (MNvit) in Chinese hamster lung (CHL) cells at Covance laboratories; Harrogate UK in support of OECD draft Test Guideline 487.

    Science.gov (United States)

    Fowler, Paul; Whitwell, James; Jeffrey, Laura; Young, Jamie; Smith, Katie; Kirkland, David

    2010-10-29

    The following genotoxic chemicals were tested in the in vitro micronucleus assay, at Covance Laboratories, Harrogate, UK in the Chinese hamster lung cell line CHL. Etoposide (a topoisomerase inhibitor), colchicine (an aneugen), mitomycin C (a DNA cross linking agent) and cyclophosphamide (an alkylating agent requiring metabolic activation) were treated with and without cytokinesis block (by addition of cytochalasin B). This work formed part of a collaborative evaluation of the toxicity measures recommended in the draft OECD Test Guideline 487 for the in vitro micronucleus test. The toxicity measures used, detecting both cytostasis and cell death, were relative population doubling, relative increase in cell counts and relative cell counts for treatments in the absence of cytokinesis block, and replication index or cytokinesis blocked proliferation index in the presence of cytokinesis block. All of the chemicals tested gave significant increases in the percentage of micronucleated cells with and without cytokinesis block at concentrations giving approximately 60% toxicity (cytostasis and cell death) or less by all of the toxicity measures used. The outcomes from this series of tests support the use of relative increase in cell counts and relative population doubling, as well as relative cell counts, as appropriate measures of cytotoxicity for the non-cytokinesis blocked in vitro micronucleus assay. Copyright © 2010 Elsevier B.V. All rights reserved.

  8. Test System Impact on System Availability

    DEFF Research Database (Denmark)

    Pau, L. F.

    1987-01-01

    The specifications are presented for an imperfect automatic test system (ATS) (test frequency distribution, reliability, false alarm rate, nondetection rate) in order to account for the availability, readiness, mean time between unscheduled repairs (MTBUR), reliability, and maintenance of the sys......The specifications are presented for an imperfect automatic test system (ATS) (test frequency distribution, reliability, false alarm rate, nondetection rate) in order to account for the availability, readiness, mean time between unscheduled repairs (MTBUR), reliability, and maintenance...... of the system subject to monitoring and test. A time-dependent Markov model is presented, and applied in three cases, with examples of numerical results provided for preventive maintenance decisions, design of an automatic test system, buffer testing in computers, and data communications....

  9. Fluidic system for long-term in vitro culturing and monitoring of organotypic brain slices

    DEFF Research Database (Denmark)

    Bakmand, Tanya; Troels-Smith, Ane R.; Dimaki, Maria

    2015-01-01

    Brain slice preparations cultured in vitro have long been used as a simplified model for studying brain development, electrophysiology, neurodegeneration and neuroprotection. In this paper an open fluidic system developed for improved long term culturing of organotypic brain slices is presented....... The positive effect of continuous flow of growth medium, and thus stability of the glucose concentration and waste removal, is simulated and compared to the effect of stagnant medium that is most often used in tissue culturing. Furthermore, placement of the tissue slices in the developed device was studied...... by numerical simulations in order to optimize the nutrient distribution. The device was tested by culturing transverse hippocampal slices from 7 days old NMRI mice for a duration of 14 days. The slices were inspected visually and the slices cultured in the fluidic system appeared to have preserved...

  10. The bacterial sealing capacity of morse taper implant-abutment systems in vitro.

    Science.gov (United States)

    Ranieri, Rogerio; Ferreira, Andreia; Souza, Emmanuel; Arcoverde, Joao; Dametto, Fabio; Gade-Neto, Cicero; Seabra, Flavio; Sarmento, Carlos

    2015-05-01

    The use of Morse taper systems in dental implantology has been associated widely with a more precise adaptation between implants and their respective abutments. This may lead to an increase in the stability of the implant system and may also prevent microbial invasion through the implant-abutment interface. The aim of this study was to investigate in vitro the ability of four commercially available Morse taper system units to impede bacterial penetration through their implant-abutment interfaces. Abutments were screwed onto the implants, and the units were subsequently immersed in Streptococcus sanguinis bacterial broth (1 × 10(8) colony forming units/mL) for 48 hours. The units were examined by scanning electron microscopy (SEM) under three conditions: 1) with the implant-abutment components assembled as units to investigate for both the existence of microgaps and the presence of bacteria; 2) with the implants and abutments separated for examination of internal surfaces; and 3) with the implant-abutment components again assembled as units to measure any microgaps detected. The mean size of the microgaps in each unit was determined by measuring, under SEM, their width in four equidistant points. Microgaps were detected in all units with no significant differences in dimension (Kruskal-Wallis test, P >0.05). Within all units, the presence of bacteria was also observed. The seals provided by the interfaces of the commercially available Morse taper implant-abutment units tested were not sufficiently small to shield the implant from bacterial penetration.

  11. Summary, the 20th quality control survey for radioisotopes in vitro tests in Japan, 1998

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1999-11-01

    For advancement of radioisotope in vitro tests such as radioimmunoassay and immunoradiometric assay, the Subcommittee for Radioisotope in vitro Test in Medical and Pharmaceutical Committee of Japan Radioisotope Association has conducted the yearly quality control survey for the test facilities in Japan since 1978. This is the summary of the 20th survey in 1998 where non-radioisotope tests like enzyme-immunoassay were involved as well. The survey was done for 143 facilities: 20 national and public university hospitals, 18 private university hospitals, 8 national hospitals, 13 public hospitals, 21 private hospitals, 41 hygienic laboratories and 22 manufacturers of reagents. Facilities examined intra- and between day-reproducibility, freeze-thaw effect and time change of the measured values on the same samples. Assays were for: growth hormone (h), somatomedin C, follicle stimulating h, luteinizing h, prolactin, thyroid stimulating h, triiodothyronines, thyroxines, thyroxine binding protein, calcitonin, insulin, C-peptide, glucagons, gastrin, testosterones, estradiol, progesterone, gonadotropin, 17{alpha}-hydroxyprogesterone, aldosterone, cortisol, dehydroepiandorosterone sulfate, renin, IgE, digoxin, {alpha}-fetoprotein, carcinoembryonic antigen, tissue polypeptide antigen, CA (125, 19-9 and 15-3), prostatic acid phosphatase, prostate specific antigen, {beta}2-microglobulin, ferritin, and neuron specific enolase. There was no great difference between this and last survey results although tendency of improvement was recognized. There were problems to be solved from the standpoint of clinical practice. (K.H.)

  12. The in vitro biokinetics of chlorpromazine and diazepam in aggregating rat brain cell cultures after repeated exposure

    NARCIS (Netherlands)

    Broeders, Jessica J W; Hermens, Joop L M; Blaauboer, Bas J; Zurich, Marie-Gabrielle

    2015-01-01

    Neurotoxic effects of compounds can be tested in vitro using cell systems. One example is aggregating rat brain cell cultures. For the extrapolation of in vitro data to the in vivo situation, it is important to take the biokinetics of the test compound into account. In addition, the exposure in vivo

  13. New in vitro system to predict chemotherapeutic efficacy of drug combinations in fresh tumor samples

    Directory of Open Access Journals (Sweden)

    Frank Christian Kischkel

    2017-03-01

    Full Text Available Background To find the best individual chemotherapy for cancer patients, the efficacy of different chemotherapeutic drugs can be predicted by pretesting tumor samples in vitro via the chemotherapy-resistance (CTR-Test®. Although drug combinations are widely used among cancer therapy, so far only single drugs are tested by this and other tests. However, several first line chemotherapies are combining two or more chemotherapeutics, leading to the necessity of drug combination testing methods. Methods We established a system to measure and predict the efficacy of chemotherapeutic drug combinations with the help of the Loewe additivity concept in combination with the CTR-test. A combination is measured by using half of the monotherapy’s concentration of both drugs simultaneously. With this method, the efficacy of a combination can also be calculated based on single drug measurements. Results The established system was tested on a data set of ovarian carcinoma samples using the combination carboplatin and paclitaxel and confirmed by using other tumor species and chemotherapeutics. Comparing the measured and the calculated values of the combination testings revealed a high correlation. Additionally, in 70% of the cases the measured and the calculated values lead to the same chemotherapeutic resistance category of the tumor. Conclusion Our data suggest that the best drug combination consists of the most efficient single drugs and the worst drug combination of the least efficient single drugs. Our results showed that single measurements are sufficient to predict combinations in specific cases but there are exceptions in which it is necessary to measure combinations, which is possible with the presented system.

  14. Correlating In Vitro Splice Switching Activity With Systemic In Vivo Delivery Using Novel ZEN-modified Oligonucleotides

    Directory of Open Access Journals (Sweden)

    Suzan M Hammond

    2014-01-01

    Full Text Available Splice switching oligonucleotides (SSOs induce alternative splicing of pre-mRNA and typically employ chemical modifications to increase nuclease resistance and binding affinity to target pre-mRNA. Here we describe a new SSO non-base modifier (a naphthyl-azo group, “ZEN™” to direct exon exclusion in mutant dystrophin pre-mRNA to generate functional dystrophin protein. The ZEN modifier is placed near the ends of a 2′-O-methyl (2′OMe oligonucleotide, increasing melting temperature and potency over unmodified 2′OMe oligonucleotides. In cultured H2K cells, a ZEN-modified 2′OMe phosphorothioate (PS oligonucleotide delivered by lipid transfection greatly enhanced dystrophin exon skipping over the same 2′OMePS SSO lacking ZEN. However, when tested using free gymnotic uptake in vitro and following systemic delivery in vivo in dystrophin deficient mdx mice, the same ZEN-modified SSO failed to enhance potency. Importantly, we show for the first time that in vivo activity of anionic SSOs is modelled in vitro only when using gymnotic delivery. ZEN is thus a novel modifier that enhances activity of SSOs in vitro but will require improved delivery methods before its in vivo clinical potential can be realized.

  15. In vitro performance investigation of bioresorbable scaffolds - Standard tests for vascular stents and beyond.

    Science.gov (United States)

    Schmidt, Wolfram; Behrens, Peter; Brandt-Wunderlich, Christoph; Siewert, Stefan; Grabow, Niels; Schmitz, Klaus-Peter

    2016-09-01

    Biodegradable polymers are the main materials for coronary scaffolds. Magnesium has been investigated as a potential alternative and was successfully tested in human clinical trials. However, it is still challenging to achieve mechanical parameters comparative to permanent bare metal (BMS) and drug-eluting stents (DES). As such, in vitro tests are required to assess mechanical parameters correlated to the safety and efficacy of the device. In vitro bench tests evaluate scaffold profiles, length, deliverability, expansion behavior including acute elastic and time-dependent recoil, bending stiffness and radial strength. The Absorb GT1 (Abbott Vascular, Temecula, CA), DESolve (Elixir Medical Corporation, Sunnyvale, CA) and the Magmaris (BIOTRONIK AG, Bülach, Switzerland) that was previously tested in the BIOSOLVE II study, were tested. Crimped profiles were 1.38±0.01mm (Absorb GT1), 1.39±0.01mm (DESolve) and 1.44±0.00mm (Magmaris) enabling 6F compatibility. Trackability was measured depending on stiffness and force transmission (pushability). Acute elastic recoil was measured at free expansion and within a mock vessel, respectively, yielding results of 5.86±0.76 and 5.22±0.38% (Absorb), 7.85±3.45 and 9.42±0.21% (DESolve) and 5.57±0.72 and 4.94±0.31% (Magmaris). Time-dependent recoil (after 1h) was observed for the Absorb and DESolve scaffolds but not for the Magmaris. The self-correcting wall apposition behavior of the DESolve did not prevent time-dependent recoil under vessel loading. The results of the suggested test methods allow assessment of technical feasibility based on objective mechanical data and highlight the main differences between polymeric and metallic bioresorbable scaffolds. Copyright © 2016 Elsevier Inc. All rights reserved.

  16. Alternative approaches for identifying acute systemic toxicity: Moving from research to regulatory testing.

    Science.gov (United States)

    Hamm, Jon; Sullivan, Kristie; Clippinger, Amy J; Strickland, Judy; Bell, Shannon; Bhhatarai, Barun; Blaauboer, Bas; Casey, Warren; Dorman, David; Forsby, Anna; Garcia-Reyero, Natàlia; Gehen, Sean; Graepel, Rabea; Hotchkiss, Jon; Lowit, Anna; Matheson, Joanna; Reaves, Elissa; Scarano, Louis; Sprankle, Catherine; Tunkel, Jay; Wilson, Dan; Xia, Menghang; Zhu, Hao; Allen, David

    2017-06-01

    Acute systemic toxicity testing provides the basis for hazard labeling and risk management of chemicals. A number of international efforts have been directed at identifying non-animal alternatives for in vivo acute systemic toxicity tests. A September 2015 workshop, Alternative Approaches for Identifying Acute Systemic Toxicity: Moving from Research to Regulatory Testing, reviewed the state-of-the-science of non-animal alternatives for this testing and explored ways to facilitate implementation of alternatives. Workshop attendees included representatives from international regulatory agencies, academia, nongovernmental organizations, and industry. Resources identified as necessary for meaningful progress in implementing alternatives included compiling and making available high-quality reference data, training on use and interpretation of in vitro and in silico approaches, and global harmonization of testing requirements. Attendees particularly noted the need to characterize variability in reference data to evaluate new approaches. They also noted the importance of understanding the mechanisms of acute toxicity, which could be facilitated by the development of adverse outcome pathways. Workshop breakout groups explored different approaches to reducing or replacing animal use for acute toxicity testing, with each group crafting a roadmap and strategy to accomplish near-term progress. The workshop steering committee has organized efforts to implement the recommendations of the workshop participants. Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. Investigation of thyroid parameters in farm animal by means of 125I in vitro tests

    International Nuclear Information System (INIS)

    Reinecke, P.; Leuthold, G.

    1988-01-01

    125 I in vitro tests especially thyroid hormone radioimmunoassays rendered it possible to study thyroidal activity of domestic animals even in large random tests. Parameters of thyroidal activity, such as effective T 4 quotient, T 3 value and total T 3 content, were investigated as to their connection to growth and environmental influence. The estimation of the hereditability yielded only low h 2 coefficients except in the T 3 value. All parameters studied depended to a great extent on farm conditions

  18. In Vitro Drug Sensitivity Tests to Predict Molecular Target Drug Responses in Surgically Resected Lung Cancer.

    Directory of Open Access Journals (Sweden)

    Ryohei Miyazaki

    Full Text Available Epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs and anaplastic lymphoma kinase (ALK inhibitors have dramatically changed the strategy of medical treatment of lung cancer. Patients should be screened for the presence of the EGFR mutation or echinoderm microtubule-associated protein-like 4 (EML4-ALK fusion gene prior to chemotherapy to predict their clinical response. The succinate dehydrogenase inhibition (SDI test and collagen gel droplet embedded culture drug sensitivity test (CD-DST are established in vitro drug sensitivity tests, which may predict the sensitivity of patients to cytotoxic anticancer drugs. We applied in vitro drug sensitivity tests for cyclopedic prediction of clinical responses to different molecular targeting drugs.The growth inhibitory effects of erlotinib and crizotinib were confirmed for lung cancer cell lines using SDI and CD-DST. The sensitivity of 35 cases of surgically resected lung cancer to erlotinib was examined using SDI or CD-DST, and compared with EGFR mutation status.HCC827 (Exon19: E746-A750 del and H3122 (EML4-ALK cells were inhibited by lower concentrations of erlotinib and crizotinib, respectively than A549, H460, and H1975 (L858R+T790M cells were. The viability of the surgically resected lung cancer was 60.0 ± 9.8 and 86.8 ± 13.9% in EGFR-mutants vs. wild types in the SDI (p = 0.0003. The cell viability was 33.5 ± 21.2 and 79.0 ± 18.6% in EGFR mutants vs. wild-type cases (p = 0.026 in CD-DST.In vitro drug sensitivity evaluated by either SDI or CD-DST correlated with EGFR gene status. Therefore, SDI and CD-DST may be useful predictors of potential clinical responses to the molecular anticancer drugs, cyclopedically.

  19. Testing strategies for embryo-fetal toxicity of human pharmaceuticals. Animal models vs. in vitro approaches: a workshop report.

    Science.gov (United States)

    van der Laan, Jan Willem; Chapin, Robert E; Haenen, Bert; Jacobs, Abigail C; Piersma, Aldert

    2012-06-01

    Reproductive toxicity testing is characterized by high animal use. For registration of pharmaceutical compounds, developmental toxicity studies are usually conducted in both rat and rabbits. Efforts have been underway for a long time to design alternatives to animal use. Implementation has lagged, partly because of uncertainties about the applicability domain of the alternatives. The reproductive cycle is complex and not all mechanisms of development can be mimicked in vitro. Therefore, efforts are underway to characterize the available alternative tests with regard to the mechanism of action they include. One alternative test is the mouse embryonic stem cell test (EST), which has been studied since the late 1990s. It is a genuine 3R "alternative" assay as it is essentially animal-free. A meeting was held to review the state-of-the-art of various in vitro models for prediction of developmental toxicity. Although the predictivity of individual assays is improving, a battery of several assays is likely to have even higher predictivity, which is necessary for regulatory acceptance. The workshop concluded that an important first step is a thorough survey of the existing rat and rabbit studies, to fully characterize the frequency of responses and the types of effects seen. At the same time, it is important to continue the optimization of in vitro assays. As more experience accumulates, the optimal conditions, assay structure, and applicability of the alternative assays are expected to emerge. Copyright © 2012 Elsevier Inc. All rights reserved.

  20. Beneficial effect of two culture systems with small groups of embryos on the development and quality of in vitro-produced bovine embryos.

    Science.gov (United States)

    Cebrian-Serrano, A; Salvador, I; Silvestre, M A

    2014-02-01

    Currently, in vitro-produced embryos derived by ovum pick up (OPU) and in vitro fertilization (IVF) technologies represent approximately one-third of the embryos worldwide in cattle. Nevertheless, the culture of small groups of embryos from an individual egg donor is an issue that OPU-IVF laboratories have to face. In this work, we tested whether the development and quality of the preimplantation embryos in vitro cultured in low numbers (five embryos) could be improved by the addition of epidermal growth factor, insulin, transferrin and selenium (EGF-ITS) or by the WOW system. With this aim, immature oocytes recovered from slaughtered heifers were in vitro matured and in vitro fertilized. Presumptive zygotes were then randomly cultured in four culture conditions: one large group (LG) (50 embryos/500 μl medium) and three smaller groups [five embryos/50 μl medium without (control) or with EGF-ITS (EGF-ITS) and five embryos per microwell in the WOW system (WOW)]. Embryos cultured in LG showed a greater ability to develop to blastocyst stage than embryos cultured in smaller groups, while the blastocyst rate of WOW group was significantly higher than in control. The number of cells/blastocyst in LG was higher than control or WOW, whereas the apoptosis rate per blastocyst was lower. On the other hand, the addition of EGF-ITS significantly improved both parameters compared to the control and resulted in similar embryo quality to LG. In conclusion, the WOW system improved embryo development, while the addition of EGF-ITS improved the embryo quality when smaller groups of embryos were cultured. © 2013 Blackwell Verlag GmbH.

  1. Sirc-cvs cytotoxicity test: an alternative for predicting rodent acute systemic toxicity.

    Science.gov (United States)

    Kitagaki, Masato; Wakuri, Shinobu; Hirota, Morihiko; Tanaka, Noriho; Itagaki, Hiroshi

    2006-10-01

    An in vitro crystal violet staining method using the rabbit cornea-derived cell line (SIRC-CVS) has been developed as an alternative to predict acute systemic toxicity in rodents. Seventy-nine chemicals, the in vitro cytotoxicity of which was already reported by the Multicenter Evaluation of In vitro Toxicity (MEIC) and ICCVAM/ECVAM, were selected as test compounds. The cells were incubated with the chemicals for 72 hrs and the IC(50) and IC(35) values (microg/mL) were obtained. The results were compared to the in vivo (rat or mouse) "most toxic" oral, intraperitoneal, subcutaneous and intravenous LD(50) values (mg/kg) taken from the RTECS database for each of the chemicals by using Pearson's correlation statistics. The following parameters were calculated: accuracy, sensitivity, specificity, prevalence, positive predictability, and negative predictability. Good linear correlations (Pearson's coefficient; r>0.6) were observed between either the IC(50) or the IC(35) values and all the LD(50) values. Among them, a statistically significant high correlation (r=0.8102, p50) values and the oral LD(50) values. By using the cut-off concentrations of 2,000 mg/kg (LD(50)) and 4,225 microg/mL (IC(50)), no false negatives were observed, and the accuracy was 84.8%. From this, it is concluded that this method could be used to predict the acute systemic toxicity potential of chemicals in rodents.

  2. In vitro test systems for the identification of gentoxic chemicals in the human environment: The proof of DNA repair synthesis in liver cells

    International Nuclear Information System (INIS)

    Rossberger, S.

    1986-01-01

    This work examines the possibilities of proving a DNA repair by gentoxic chemicals in primary hepatozytes and 2sFou liver cells of rates. Two different processes used for the in vitro mutagenic testing of alien substances for determining the DNA repair synthesis in primary hepatozytes, in the autoradiographic method and the gradient centrifuging method, are compared regarding their reliability and sensitivity. The rat hepatom cell line 2sFou was examined for its suitability for proving chemically induced DNA repair, instead of primary hepatozytes. (orig./MG) [de

  3. In vitro susceptibility testing of Aspergillus spp. against voriconazole, itraconazole, posaconazole, amphotericin B and caspofungin.

    Science.gov (United States)

    Shi, Jun-yan; Xu, Ying-chun; Shi, Yi; Lü, Huo-xiang; Liu, Yong; Zhao, Wang-sheng; Chen, Dong-mei; Xi, Li-yan; Zhou, Xin; Wang, He; Guo, Li-na

    2010-10-01

    During recent years, the incidence of serious infections caused by opportunistic fungi has increased dramatically due to alterations of the immune status of patients with hematological diseases, malignant tumors, transplantations and so forth. Unfortunately, the wide use of triazole antifungal agents to treat these infections has lead to the emergence of Aspergillus spp. resistant to triazoles. The present study was to assess the in vitro activities of five antifungal agents (voriconazole, itraconazole, posaconazole, amphotericin B and caspofungin) against different kinds of Aspergillus spp. that are commonly encountered in the clinical setting. The agar-based Etest MIC method was employed. One hundred and seven strains of Aspergillus spp. (5 species) were collected and prepared according to Etest Technique Manuel. Etest MICs were determined with RPMI agar containing 2% glucose and were read after incubation for 48 hours at 35°C. MIC(50), MIC(90) and MIC range were acquired by Whonet 5.4 software. The MIC(90) of caspofungin against A. fumigatus, A. flavus and A. nidulans was 0.094 µg/ml whereas the MIC(90) against A. niger was 0.19 µg/ml. For these four species, the MIC(90) of caspofungin was the lowest among the five antifungal agents. For A. terrus, the MIC(90) of posaconazole was the lowest. For A. fumigatus and A. flavus, the MIC(90) in order of increasing was caspofungin, posaconazole, voriconazole, itraconazole, and amphotericin B. The MIC of amphotericin B against A. terrus was higher than 32 µg/ml in all 7 strains tested. The in vitro antifungal susceptibility test shows the new drug caspofungin, which is a kind of echinocandins, has good activity against the five species of Aspergillus spp. and all the triazoles tested have better in vitro activity than traditional amphotericin B.

  4. Expert consensus on an in vitro approach to assess pulmonary fibrogenic potential of aerosolized nanomaterials.

    Science.gov (United States)

    Clippinger, Amy J; Ahluwalia, Arti; Allen, David; Bonner, James C; Casey, Warren; Castranova, Vincent; David, Raymond M; Halappanavar, Sabina; Hotchkiss, Jon A; Jarabek, Annie M; Maier, Monika; Polk, William; Rothen-Rutishauser, Barbara; Sayes, Christie M; Sayre, Phil; Sharma, Monita; Stone, Vicki

    2016-07-01

    The increasing use of multi-walled carbon nanotubes (MWCNTs) in consumer products and their potential to induce adverse lung effects following inhalation has lead to much interest in better understanding the hazard associated with these nanomaterials (NMs). While the current regulatory requirement for substances of concern, such as MWCNTs, in many jurisdictions is a 90-day rodent inhalation test, the monetary, ethical, and scientific concerns associated with this test led an international expert group to convene in Washington, DC, USA, to discuss alternative approaches to evaluate the inhalation toxicity of MWCNTs. Pulmonary fibrosis was identified as a key adverse outcome linked to MWCNT exposure, and recommendations were made on the design of an in vitro assay that is predictive of the fibrotic potential of MWCNTs. While fibrosis takes weeks or months to develop in vivo, an in vitro test system may more rapidly predict fibrogenic potential by monitoring pro-fibrotic mediators (e.g., cytokines and growth factors). Therefore, the workshop discussions focused on the necessary specifications related to the development and evaluation of such an in vitro system. Recommendations were made for designing a system using lung-relevant cells co-cultured at the air-liquid interface to assess the pro-fibrogenic potential of aerosolized MWCNTs, while considering human-relevant dosimetry and NM life cycle transformations. The workshop discussions provided the fundamental design components of an air-liquid interface in vitro test system that will be subsequently expanded to the development of an alternative testing strategy to predict pulmonary toxicity and to generate data that will enable effective risk assessment of NMs.

  5. Procedure for implementing the system of quality management in the testing laboratory of the Center for Genetic Engineering and Biotechnology in Sancti Spiritus.

    Directory of Open Access Journals (Sweden)

    Lídice Peraza Cruz

    2014-03-01

    Full Text Available Implementation of Quality Management System in testing laboratories offers the possibility of its accreditation and a frame for cooperation with other organizations, supporting information and experience exchange, as well as standards and procedures harmonization. To improve the performance of the Center for Genetic Engineering and Biotechnology of Sancti Spíritus testing laboratory, assuring technically valid data and results which promote technical competence and credibility of in vitro diagnostics and biological reagents products, a procedure was designed to implement a Quality Management System. This procedure applies Deming´s Quality Cycle and considers all relevant requirements in NC ISO/IEC 17025:2006 “General requirements for the competence of testing and calibration laboratories” and Regulation No. 20 2004 “Good Manufacturing Practices for in vitro Diagnostics” of Center for State Control of Drugs, Equipment and Medical Devices. We recommend an auto evaluation method, designed by authors, to verify quality management system accomplishment.

  6. In vitro tests for aerosol deposition II: IVIVCs for different dry powder inhalers in normal adults.

    Science.gov (United States)

    Delvadia, Renishkumar; Hindle, Michael; Longest, P Worth; Byron, Peter R

    2013-06-01

    A new in vitro test method for dry powder inhalers (DPIs) was recently found to be predictive of the published in vivo results for Budelin Novolizer. The present study was intended to assess the method's robustness by evaluating correlations between average drug deposition in vitro and in vivo from five different DPIs. In vitro drug deposition from five marketed DPIs was assessed in a realistic physical airway model of a "medium" sized adult in an experimental setup that allowed deposition to be characterized regionally for carefully selected simulated air flow rate versus time profiles. The DPIs studied were Spiriva(®) HandiHaler(®), Relenza(®) Diskhaler(®), Salbutamol Easyhaler(®), Pulmicort(®) Turbuhaler(®), and Foradil(®) Aerolizer(®). In vitro regional deposition results were compared with those reported in the literature in order to create in vitro-in vivo correlations (IVIVCs) for each inhaler. Mean percent total lung deposition (TLD ± SD) in vitro for Spiriva HandiHaler, Relenza Diskhaler, Salbutamol Easyhaler, Pulmicort Turbuhaler, and Foradil Aerolizer were 17.3 ± 1.2, 22.6 ± 1.1, 29.0 ± 1.1, 28.0 ± 3.0, and 21.7 ± 1.2, respectively. These results showed excellent agreement with reported in vivo values, with absolute prediction errors in TLD of ≤ 2% for all DPIs except Relenza Diskhaler. Similarly, in vitro mouth-throat and device deposition results were stoichiometrically comparable to those reported in vivo for all DPIs except Relenza Diskhaler and Turbuhaler. Inspection of the scintigraphy studies for Relenza Diskhaler and Turbohaler revealed possible problems with powder labeling and result interpretation in their in vivo clinical assessments. A characteristic physical airway model representing a medium-sized adult, when coupled to carefully chosen characteristic inhalation maneuvers used in the clinic, produced results that correlated with regional drug deposition estimates from scintigraphy across a group of different DPIs.

  7. In vitro meat production system: why and how?

    Science.gov (United States)

    Sharma, Shruti; Thind, Sukhcharanjit Singh; Kaur, Amarjeet

    2015-12-01

    Due to the nutritional importance and the sustained popularity of meat as a foodstuff, the livestock production sector has been expanding incessantly. This exponential growth of livestock meat sector poses a gigantic challenge to the sustainability of food production system. A new technological breakthrough is being contemplated to develop a substitute for livestock meat. The idea is to grow meat in a culture in the lab and manipulate its composition selectively. This paper aims to discuss the concept of In Vitro Meat production system, articulate the underlying technology and analyse the context of its implications, as proposed by several scientists and stakeholders. The challenges facing this emerging technology have also been discussed.

  8. Surface enhanced Raman spectroscopy (SERS for in vitro diagnostic testing at the point of care

    Directory of Open Access Journals (Sweden)

    Marks Haley

    2017-06-01

    Full Text Available Point-of-care (POC device development is a growing field that aims to develop low-cost, rapid, sensitive in-vitro diagnostic testing platforms that are portable, self-contained, and can be used anywhere – from modern clinics to remote and low resource areas. In this review, surface enhanced Raman spectroscopy (SERS is discussed as a solution to facilitating the translation of bioanalytical sensing to the POC. The potential for SERS to meet the widely accepted “ASSURED” (Affordable, Sensitive, Specific, User-friendly, Rapid, Equipment-free, and Deliverable criterion provided by the World Health Organization is discussed based on recent advances in SERS in vitro assay development. As SERS provides attractive characteristics for multiplexed sensing at low concentration limits with a high degree of specificity, it holds great promise for enhancing current efforts in rapid diagnostic testing. In outlining the progression of SERS techniques over the past years combined with recent developments in smart nanomaterials, high-throughput microfluidics, and low-cost paper diagnostics, an extensive number of new possibilities show potential for translating SERS biosensors to the POC.

  9. Surface enhanced Raman spectroscopy (SERS) for in vitro diagnostic testing at the point of care

    Science.gov (United States)

    Marks, Haley; Schechinger, Monika; Garza, Javier; Locke, Andrea; Coté, Gerard

    2017-06-01

    Point-of-care (POC) device development is a growing field that aims to develop low-cost, rapid, sensitive in-vitro diagnostic testing platforms that are portable, self-contained, and can be used anywhere - from modern clinics to remote and low resource areas. In this review, surface enhanced Raman spectroscopy (SERS) is discussed as a solution to facilitating the translation of bioanalytical sensing to the POC. The potential for SERS to meet the widely accepted "ASSURED" (Affordable, Sensitive, Specific, User-friendly, Rapid, Equipment-free, and Deliverable) criterion provided by the World Health Organization is discussed based on recent advances in SERS in vitro assay development. As SERS provides attractive characteristics for multiplexed sensing at low concentration limits with a high degree of specificity, it holds great promise for enhancing current efforts in rapid diagnostic testing. In outlining the progression of SERS techniques over the past years combined with recent developments in smart nanomaterials, high-throughput microfluidics, and low-cost paper diagnostics, an extensive number of new possibilities show potential for translating SERS biosensors to the POC.

  10. In Vitro Antimicrobial Susceptibility Testing of Animal Nocardia Isolated from Field Cases of Skin Diseases

    Directory of Open Access Journals (Sweden)

    M. A. Oyekunle

    2001-03-01

    Full Text Available In vitro antimicrobial tests were carried out on strains of Nocardia isolated from field cases of cutaneous nocardiosis in farm animals. Results with the disc diffusion test showed the multiresistant nature of the isolates, but 23.81 and 21.43% were sensitive to ciprofloxacin and gentamycin, respectively. The MIC mode and range for oxytetracycline were 12.5 and 3.12–25 μg/ml, respectively, while those of erythromycin were 3.12 and 0.78–6.25 μg/ml, respectively.

  11. Boosting Immune Responses Against Bacterial Pathogens: In Vitro Analysis of Immunomodulators (In Vitro Analyse van de Stimulerende Werking van Verschillende Stoffen op het Immuunsysteem)

    National Research Council Canada - National Science Library

    Kleij, D. van der

    2007-01-01

    .... Three potential broad-spectrum therapeutics (MPL, MDP and ssPolyU) and their combinations were tested in an in vitro dendritic cell culture system, since dendritic cells play a central role in the development of immune...

  12. In vitro cytotoxicity testing of Ubiquicidin 29-41-99mTc

    International Nuclear Information System (INIS)

    Ocampo, Ivette Z.; Okazaki, Kayo; Dias, Luis Alberto Pereira; Higa, Olga Z.; Silva, Fabiana M. da; Vieira, Daniel P.; Passos, Priscila; Esteves-Pedro, Natalia M.

    2015-01-01

    The work carried out cytotoxicity tests using a radiopharmaceutical compound produced at IPEN/CNEN-SP to certify its safety through in vitro cytotoxicity tests. Since 2009, the Brazilian regulatory agency (ANVISA) requires that such tests have to be carried out following good laboratory practices (GLP) and in according to the OECD (Organisation for Economic Co-operation and Development) guidelines in order to certify its safety for medical use. Those guidelines comprises series of technical recommendations performed to assure quality of experiments. The study chose Ubiquicidin 29-41, an antimicrobial peptide used to discriminate bacterial infection foci from inflammatory sites. Amounts of UBI 29-41 were conjugated or not to 99m Tc and diluted to equivalent concentrations of 10, 100 and 1000% of the maximum dose (or activity) administered in adults. Possible cytotoxic effects were evaluated in comparison to untreated controls as well as positive and negative damage controls. Both full (radioactive) radiopharmaceuticals, as their precursors (only molecules without conjugation to isotopes) showed no significant cytotoxic effect (cytotoxicity ≤ 10%). The study was conducted for the first time in the country comprising preclinical testing of this radiopharmaceutical in accordance with internationally accepted quality parameters, ensuring the safety of its use and enabling inclusion in the pharmaceutical regulatory agenda. (author)

  13. The development of in vitro mutagenicity testing systems using T-lymphocytes: Progress report, November 1, 1987--May 31, 1988

    International Nuclear Information System (INIS)

    Albertini, R.J.

    1988-05-01

    We have investigated the mutagenic effects of ionizing irradiations in human T-lymphocytes. These have included in vitro exposure of G 0 phase peripheral blood T-lymphocytes to gamma irradiation, with subsequent growth for phenotypic expression and selection of 6-thioguanine resistant (TG/sup r/) colonies to measure mutation induction at the hprt mutations. We have studied the DNA alterations in both in vivo and in vitro induced mutants by Southern blot analysis with an hprt cDNA probe in order to investigate the possible specificity of radiation damage. We have employed Southern blot analyses with T-cell receptor (TCR) gene probes in order to define the clonality of these mutants and allow definition of the mutation frequency underlying the measured mutant frequency. This approach has allowed us to define the molecular nature of mutations induced both in vivo and in vitro by ionizing radiation. Southern blot analysis of 70 in vitro mutant clones induced by 300 rads of gamma irradiation have shown them to be the result of 24 independent mutations by TCR gene rearrangement patterns. Eighteen (0.66) of these were found to have large hprt DNA alterations. Individuals therapeutically exposed to ionizing irradiations show higher TG/sup r/ mutant frequencies than those found in normal controls and a higher fraction of these mutants also contain hprt gene alterations detected by Southern blot analysis (0.34 vs. 0.10--0.15 in controls, respectively). The fraction of mutations which contain detectable DNA alterations is thus elevated after both in vitro and in vivo irradiation of the cells. Analysis of breakpoints in these mutants has allowed us to estimate the maximum size deletion recoverable at the hprt locus. 8 refs., 5 figs., 4 tabs

  14. Development of an in vitro test system for assessment of male, reproductive toxicity.

    Science.gov (United States)

    Habas, Khaled; Anderson, Diana; Brinkworth, Martin

    2014-02-10

    There is a need for improved reproductive toxicology assays that do not require large numbers of animals but are sensitive and informative. Therefore, Staput velocity-sedimentation separation followed by culture of specific mouse testicular cells was used as such a system. The specificity of separation was assessed using immunocytochemistry to identify spermatids, spermatocytes and spermatogonia. The efficacy of the system to detect toxicity was then evaluated by analysing the effects of hydrogen peroxide (H2O2) by the terminal uridine-deoxynucleotide end-labelling (TUNEL) assay to show the rate of apoptosis induced among the different types of germ cells. We found that 2 h of treatment at both 1 and 10 μM induced increases of over ∼10-fold in the percentage of apoptotic cells (p≤0.001), confirming that testicular germ cells are prone to apoptosis at very low concentrations of H2O2. It was also demonstrated for the first time for this compound that spermatogonia are significantly more susceptible than spermatocytes, which are more affected than spermatids. This reflects the proportion of actively dividing cells in these cell types, suggesting a mechanism for the differential sensitivity. The approach should thus form the basis of a useful test system for reproductive and genetic toxicology in the future. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

  15. In vitro testing of daptomycin plus rifampin againstmethicillin-resistant Staphylococcus aureus resistant to rifampin

    International Nuclear Information System (INIS)

    Khaswneh, Faisal A.; Ashcraft, Deborah S.; Pankey, George A.

    2008-01-01

    Objective was to test for synergy between daptomycin (DAP) and rifampin(RIF) against RIF-resistant methicillin-resistant Staphylococcus aureus(MRSA) isolates. Synergy testing using time-kill assay (TKA) was performed on6 clinically and genetically unique RIF-resistant MRSA isolates. The isolateswere identified out of 489 (1.2%) samples collected during April 2003 toAugust 2006, from patients at the Ochsner Medical Center in New Orleans,Louisiana, United States of America. Synergy testing of DAP plus RIF by TKAshowed that 5 isolates were different, but one isolate was antagonistic. Ourin-vitro study failed to demonstrate synergy between DAP plus RIF, againstour RIF-resistant MRSA isolates. Clinical failure of this combination shouldprompt the clinician to consider antagonism as one of the potential causes.(author)

  16. Micro- and nano bio-based delivery systems for food applications: In vitro behavior.

    Science.gov (United States)

    de Souza Simões, Lívia; Madalena, Daniel A; Pinheiro, Ana C; Teixeira, José A; Vicente, António A; Ramos, Óscar L

    2017-05-01

    Micro- and nanoencapsulation is an emerging technology in the food field that potentially allows the improvement of food quality and human health. Bio-based delivery systems of bioactive compounds have a wide variety of morphologies that influence their stability and functional performance. The incorporation of bioactive compounds in food products using micro- and nano-delivery systems may offer extra health benefits, beyond basic nutrition, once their encapsulation may provide protection against undesired environmental conditions (e.g., heat, light and oxygen) along the food chain (including processing and storage), thus improving their bioavailability, while enabling their controlled release and target delivery. This review provides an overview of the bio-based materials currently used for encapsulation of bioactive compounds intended for food applications, as well as the main production techniques employed in the development of micro- and nanosystems. The behavior of such systems and of bioactive compounds entrapped into, throughout in vitro gastrointestinal systems, is also tracked in a critical manner. Comparisons between various in vitro digestion systems (including the main advantages and disadvantages) currently in use, as well as correlations between the behavior of micro- and nanosystems studied through in vitro and in vivo systems were highlighted and discussed here for the first time. Finally, examples of bioactive micro- and nanosystems added to food simulants or to real food matrices are provided, together with a revision of the main challenges for their safe commercialization, the regulatory issues involved and the main legislation aspects. Copyright © 2017 Elsevier B.V. All rights reserved.

  17. Testing pollen of single and stacked insect-resistant Bt-maize on in vitro reared honey bee larvae.

    Science.gov (United States)

    Hendriksma, Harmen P; Härtel, Stephan; Steffan-Dewenter, Ingolf

    2011-01-01

    The ecologically and economic important honey bee (Apis mellifera) is a key non-target arthropod species in environmental risk assessment (ERA) of genetically modified (GM) crops. Honey bee larvae are directly exposed to transgenic products by the consumption of GM pollen. But most ERA studies only consider responses of adult bees, although Bt-proteins primarily affect the larval phases of target organisms. We adopted an in vitro larvae rearing system, to assess lethal and sublethal effects of Bt-pollen consumption in a standardized eco-toxicological bioassay. The effects of pollen from two Bt-maize cultivars, one expressing a single and the other a total of three Bt-proteins, on the survival and prepupae weight of honey bee larvae were analyzed. The control treatments included pollen from three non-transgenic maize varieties and of Heliconia rostrata. Three days old larvae were fed the realistic exposure dose of 2 mg pollen within the semi-artificial diet. The larvae were monitored over 120 h, until the prepupal stage, where larvae terminate feeding and growing. Neither single nor stacked Bt-maize pollen showed an adverse effect on larval survival and the prepupal weight. In contrast, feeding of H. rostrata pollen caused significant toxic effects. The results of this study indicate that pollen of the tested Bt-varieties does not harm the development of in vitro reared A. mellifera larvae. To sustain the ecosystem service of pollination, Bt-impact on A. mellifera should always be a crucial part of regulatory biosafety assessments. We suggest that our approach of feeding GM pollen on in vitro reared honey bee larvae is well suited of becoming a standard bioassay in regulatory risk assessments schemes of GM crops.

  18. Problems in extrapolating genotoxicity data from cellular systems in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Mohn, G.R.; van Zeeland, A.A.

    1986-01-01

    The experiments involved quantitative comparative mutagenesis determinations using ethylating agents differing in DNA reactivity, mammalian metabolism, and mutagenic potency. The results can be summarized as follows: (1) Under standardized in vitro treatment conditions and at identical DNA dose levels, the frequencies of induced mutations observed in the present bacterial systems are not representative and usually lower than those induced in cultured mammalian cells. (2) Under the same in vitro treatment conditions, the relative mutagenic potency order is identical in bacteria and the mammalian cells, namely, N-ethyl-N'-nitrosoquinioline(ENNG) > ethylnitrosourea (ENU) > diethylsulfate (DES) > ethylmethanesulfonate (EMS); diethylnitrosamine (DEN) being not mutagenic in the two organisms without addition of mammalian (rodent) liver preparations containing Cyt-P450-dependent mixed function oxidases. (3) The extremely high mutagenic activity of ENNG in bacteria and mammalian cells in vitro is very probably due to its intracellular, glutathione-mediated activation to highly reactive chemical species. (4) Both in bacteria and the mammalian cells the frequencies of induced mutations are directly proportional to the levels of O6-ethylguanine formed in DNA after exposure to ENNG, ENU, DES, and EMS, in contrast to results obtained when other DNA adducts are used as dose parameters. (5) The addition of mouse liver S-9 preparations in the in vitro assays allows to determine which chemicals are likely to be substantially activated (DEN) or deactivated (ENNG) in vivo; however, the mutagenic potency order remained the same under these conditions. (6) When the indicator bacteria are exposed directly to the in vivo metabolism of living animals, the relative order of mutagenic potency on the basis of exposure levels becomes drastically different from that observed in vitro, using either E. coli or V79 mammalian cells.

  19. In vitro anti-proliferative effect of interferon alpha in solid tumors: A potential predicative test

    International Nuclear Information System (INIS)

    Fuchsberger, N.; Kubes, M.; Kontsek, P.; Borecky, L.; Hornak, M.; Silvanova; Godal, A.; Svec, J.

    1993-01-01

    An in vitro test for the anti-proliferative effect of human leukocyte interferon (IFN-alpha) was performed in primary cultures of tumor cells obtained from 32 patients with either malignant melanoma (13), renal carcinoma (4) or bladder carcinoma (15). Our results demonstrated activity of IFN in all three groups of solid tumors. However, appreciable differences in sensitivity to anti-proliferative effect of IFN between individual tumors of the same type were found. The potential of this anti-proliferative test for prediction of treatment response in IFN-therapy is discussed. (author)

  20. Determination of the potency of a novel saw palmetto supercritical CO2 extract (SPSE) for 5α-reductase isoform II inhibition using a cell-free in vitro test system.

    Science.gov (United States)

    Pais, Pilar; Villar, Agustí; Rull, Santiago

    2016-01-01

    The nicotinamide adenine dinucleotide phosphate-dependent membrane protein 5α-reductase catalyses the conversion of testosterone to the most potent androgen - 5α-dihydrotestosterone. Two 5α-reductase isoenzymes are expressed in humans: type I and type II. The latter is found primarily in prostate tissue. Saw palmetto extract (SPE) has been used extensively in the treatment of lower urinary tract symptoms secondary to benign prostatic hyperplasia (BPH). The pharmacological effects of SPE include the inhibition of 5α-reductase, as well as anti-inflammatory and antiproliferative effects. Clinical studies of SPE have been inconclusive - some have shown significant results, and others have not - possibly the result of varying bioactivities of the SPEs used in the studies. To determine the in vitro potency in a cell-free test system of a novel SP supercritical CO2 extract (SPSE), an inhibitor of the 5α-reductase isoenzyme type II. The inhibitory potency of SPSE was compared to that of finasteride, an approved 5α-reductase inhibitor, on the basis of the enzymatic conversion of the substrate androstenedione to the 5α-reduced product 5α-androstanedione. By concentration-dependent inhibition of 5α-reductase type II in vitro (half-maximal inhibitory concentration 3.58±0.05 μg/mL), SPSE demonstrated competitive binding toward the active site of the enzyme. Finasteride, the approved 5α-reductase inhibitor tested as positive control, led to 63%-75% inhibition of 5α-reductase type II. SPSE effectively inhibits the enzyme that has been linked to BPH, and the amount of extract required for activity is comparatively low. It can be confirmed from the results of this study that SPSE has bioactivity that promotes prostate health at a level that is superior to that of many other phytotherapeutic extracts. The bioactivity of SPSE corresponds favorably to that reported for the hexane extract used in a large number of positive BPH clinical trials, as well as to finasteride

  1. Labelled T{sub 3}, T{sub 4} and TBP for In Vitro Testing of Thyroid Function in Man and Animals

    Energy Technology Data Exchange (ETDEWEB)

    Czerniak, P.; Boruchowski, Sabina; Shomron, I. [Dept. of Radiotherapy and Isotopes, Tel-Hashomer Hospital, Tel-Aviv University Medical School, Faculty of Continuing Medical Education, Tel-Aviv (Israel)

    1970-02-15

    Iodothyronines are bound to determined electrophoretic fractions of serum proteins - TBP (TBG, TBA, TBPA). Radioiodine labelled T{sub 3} and T{sub 4} complex the free TBP fractions until saturation. The excess of added in vitro thyronines is then absorbed by the RBC. The changes described can be detected and quantitatively determined by radioisotope tests: radioelectrophoresis - T{sub 3/4} BP test, and RBC - {sup 125}I T{sub 3} test (Hamolsky test). The in vitro tests of the thyroid function can be clinically reliable if the protein fractions are normal, and they may be altered without thyroid pathology if the TBG fractions are abnormal. The electrophoretic fractions vary quantitatively and qualitatively in animals of different classes and orders. We chose these features to study the correlation between iodothyronines, plasma proteins and the above-mentioned thyroid tests. Twenty-two animal species (arranged according to the increasing percentage of the Hamolsky test) were examined: goat (8.6%, cow, lamb, calf, man, camel, goose, hamster, rat, turkey, marmot, duck, horse, donkey, hen, dog, pigeon, rabbit, guinea-pig, mouse, fish and frog (91.5%). The following additional parameters were examined: PBI, serum quantitative electrophoresis, T{sub 3} BP studies. All the tests were performed under identical technical conditions. Results and conclusions: (1) The Hamolsky test in the examined animals ranges from 9% to 92%. It is highest in the poicolothermics, in which much prealbumin and few glubulins are found on electrophoresis. The T{sub 3} BP is low, and about a half of the added {sup 125}I T{sub 3} remains unbound. (2) T{sub 3} and T{sub 4} are complexed with T{sub 3} BP and T{sub 4} BP fractions, which correspond to prealbumin, albumin, alpha 1-2, beta and exceptionally gamma globulin. The fractions are variable but characteristic for each animal species. T{sub 3} BP does not correlate exactly with T{sub 4} BP, and seems to be distributed over more fractions

  2. A simple in vitro test tube method for estimating the bioavailability of phosphorus in feed ingredients for swine.

    Science.gov (United States)

    Bollinger, David W; Tsunoda, Atsushi; Ledoux, David R; Ellersieck, Mark R; Veum, Trygve L

    2004-04-07

    A simplified in vitro test tube (TT) method was developed to estimate the percentage of available P in feed ingredients for swine. The entire digestion procedure with the TT method consists of three consecutive enzymatic digestions carried out in a 50-mL conical test tube: (1) Pre-digestion with endo-xylanase and beta-glucanase for 1 h, (2) peptic digestion for 2 h, and (3) pancreatic digestion for 2 or 4 h. The TT method is simpler and much easier to perform compared to the dialysis tubing (DT) method, because dialysis tubing is not used. Reducing sample size from 1.0 to 0.25 g for the TT method improved results. In conclusion, the accuracy and validity of the TT method is equal to that of our more complicated DT method (r = 0.97, P < 0.001), designed to mimic the digestive system of swine, for estimating the availability of P in plant-origin feed ingredients.

  3. Test report - caustic addition system operability test procedure

    International Nuclear Information System (INIS)

    Parazin, R.E.

    1995-01-01

    This Operability Test Report documents the test results of test procedure WHC-SD-WM-OTP-167 ''Caustic Addition System Operability Test Procedure''. The Objective of the test was to verify the operability of the 241-AN-107 Caustic Addition System. The objective of the test was met

  4. In vitro propagation of avocado (Persea drymifolia Ness.)

    International Nuclear Information System (INIS)

    Barrera-Guerra, J.L.; Ramirez-Malagon, R.; Martinez-Jaime, O.A.

    2001-01-01

    In the past 20 years, reports on micropropagation and rooting in vitro of avocado shoots, with diverse origins and treatments, have been published. However, none of them reached the level required for large scale propagation of the species. It is considered that, in the first place, the micropropagation of avocado requires an efficient system of rooting. Therefore, a system to induce the rooting in vitro of avocado shoots, based on indole-3-butyric acid (IBA) pulses and some treatments based on thidiazuron (TDZ) was tested, using 40 explants per treatment. The treatments with TDZ did not succeed in rooting shoots. Some treatments with pulses of IBA induced the following rooting results: without growth regulators, 16%; 4,000 mg L -1 of IBA for 5 seconds, 8.3% rooted; 100 mg L -1 for 72 hours, 20%; 50 mg L -1 for 72 hours, 15.4%; 150 mg L -1 for 24 hours, 5% rooted. It is considered possible to improve these results by adjusting the range of IBA concentrations as well as in the time range of pulse applications. Finally, it is easier to establish in vitro explants derived from mature seeds or embryos germinated in vitro. (author)

  5. Validity of in vitro tests on aqueous spray pumps as surrogates for nasal deposition, absorption, and biologic response.

    Science.gov (United States)

    Suman, Julie D; Laube, Beth L; Dalby, Richard

    2006-01-01

    This research investigated the impact of the full range of in vitro spray characterization tests described in the FDA Draft Bioequivalence Guidance on nasal deposition pattern, pharmacokinetics, and biological response to nicotine administered by two aqueous nasal spray pumps in human volunteers. Nicotine was selected as a model drug (even though it is not locally acting) based on its ability to alter cardiac function and available plasma assay. Significant differences in pump performance-including mean volume diameters, spray angle, spray width, and ovality ratios-were observed between the two pumps. There were no significant differences in deposition pattern, or pharmacokinetic or pharmacodynamic response to the nasally administered nicotine. Although there were statistical differences in the in vitro tests between the two pumps, these differences did not result in significant alterations in the site of droplet deposition within the nose, the rate and extent of nicotine absorption, or the physiologic response it induced. These results suggest that current measures of in vitro performance, particularly spray angle and spray pattern (ovality), may not be clinically relevant. Additional research is needed to define what spray pump characteristics are likely to produce differences in deposition pattern and drug response.

  6. SPECTR System Operational Test Report

    International Nuclear Information System (INIS)

    Landman, W.H. Jr.

    2011-01-01

    This report overviews installation of the Small Pressure Cycling Test Rig (SPECTR) and documents the system operational testing performed to demonstrate that it meets the requirements for operations. The system operational testing involved operation of the furnace system to the design conditions and demonstration of the test article gas supply system using a simulated test article. The furnace and test article systems were demonstrated to meet the design requirements for the Next Generation Nuclear Plant. Therefore, the system is deemed acceptable and is ready for actual test article testing.

  7. Dissolving polyvinylpyrrolidone-based microneedle systems for in-vitro delivery of sumatriptan succinate.

    Science.gov (United States)

    Ronnander, P; Simon, L; Spilgies, H; Koch, A; Scherr, S

    2018-03-01

    In-vitro permeation studies were conducted to assess the feasibility of fabricating dissolving-microneedle-array systems to release sumatriptan succinate. The formulations consisted mainly of the encapsulated active ingredient and a water-soluble biologically compatible polymer, polyvinylpyrrolidone (PVP), approved by the U.S. Food and Drug Administration (FDA). Tests with Franz-type diffusion cells and Göttingen minipig skins showed an increase of the transdermal flux compared to passive diffusion. A preparation, containing 30% by mass of PVP and 8.7mg sumatriptan, produced a delivery rate of 395±31μg/cm 2 h over a 7-hour period after a negligible lag time of approximately 39min. Theoretically, a 10.7cm 2 microneedle-array patch loaded with 118.8mg of the drug would provide the required plasma concentration, 72ng/mL, for nearly 7h. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. 21 CFR 864.5425 - Multipurpose system for in vitro coagulation studies.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Multipurpose system for in vitro coagulation studies. 864.5425 Section 864.5425 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Automated and Semi-Automated...

  9. The in vitro and in vivo validation of a mobile non-contact camera-based digital imaging system for tooth colour measurement.

    Science.gov (United States)

    Smith, Richard N; Collins, Luisa Z; Naeeni, Mojgan; Joiner, Andrew; Philpotts, Carole J; Hopkinson, Ian; Jones, Clare; Lath, Darren L; Coxon, Thomas; Hibbard, James; Brook, Alan H

    2008-01-01

    To assess the reproducibility of a mobile non-contact camera-based digital imaging system (DIS) for measuring tooth colour under in vitro and in vivo conditions. One in vitro and two in vivo studies were performed using a mobile non-contact camera-based digital imaging system. In vitro study: two operators used the DIS to image 10 dry tooth specimens in a randomised order on three occasions. In vivo study 1:25 subjects with two natural, normally aligned, upper central incisors had their teeth imaged using the DIS on four consecutive days by one operator to measure day-to-day variability. On one of the four test days, duplicate images were collected by three different operators to measure inter- and intra-operator variability. In vivo study 2:11 subjects with two natural, normally aligned, upper central incisors had their teeth imaged using the DIS twice daily over three days within the same week to assess day-to-day variability. Three operators collected images from subjects in a randomised order to measure inter- and intra-operator variability. Subject-to-subject variability was the largest source of variation within the data. Pairwise correlations and concordance coefficients were > 0.7 for each operator, demonstrating good precision and excellent operator agreement in each of the studies. Intraclass correlation coefficients (ICCs) for each operator indicate that day-to-day reliability was good to excellent, where all ICC's where > 0.75 for each operator. The mobile non-contact camera-based digital imaging system was shown to be a reproducible means of measuring tooth colour in both in vitro and in vivo experiments.

  10. In vitro performance investigation of bioresorbable scaffolds – Standard tests for vascular stents and beyond

    Energy Technology Data Exchange (ETDEWEB)

    Schmidt, Wolfram, E-mail: wolfram.schmidt@uni-rostock.de [Institute for Biomedical Engineering, University Medicine Rostock, Friedrich-Barnewitz-Strasse 4, D-18119 Rostock-Warnemünde (Germany); Behrens, Peter, E-mail: peter.behrens@uni-rostock.de [Institute for Biomedical Engineering, University Medicine Rostock, Friedrich-Barnewitz-Strasse 4, D-18119 Rostock-Warnemünde (Germany); Brandt-Wunderlich, Christoph, E-mail: christoph.brandt@uni-rostock.de [Institute for ImplantTechnology and Biomaterials – IIB e.V., Associated Institute of the University of Rostock, Friedrich-Barnewitz-Strasse 4, D-18119 Rostock-Warnemünde (Germany); Siewert, Stefan, E-mail: stefan.siewert@uni-rostock.de [Institute for ImplantTechnology and Biomaterials – IIB e.V., Associated Institute of the University of Rostock, Friedrich-Barnewitz-Strasse 4, D-18119 Rostock-Warnemünde (Germany); Grabow, Niels, E-mail: niels.grabow@uni-rostock.de [Institute for Biomedical Engineering, University Medicine Rostock, Friedrich-Barnewitz-Strasse 4, D-18119 Rostock-Warnemünde (Germany); Schmitz, Klaus-Peter, E-mail: klaus-peter.schmitz@uni-rostock.de [Institute for ImplantTechnology and Biomaterials – IIB e.V., Associated Institute of the University of Rostock, Friedrich-Barnewitz-Strasse 4, D-18119 Rostock-Warnemünde (Germany)

    2016-09-15

    Background/Purpose: Biodegradable polymers are the main materials for coronary scaffolds. Magnesium has been investigated as a potential alternative and was successfully tested in human clinical trials. However, it is still challenging to achieve mechanical parameters comparative to permanent bare metal (BMS) and drug-eluting stents (DES). As such, in vitro tests are required to assess mechanical parameters correlated to the safety and efficacy of the device. Methods/Materials: In vitro bench tests evaluate scaffold profiles, length, deliverability, expansion behavior including acute elastic and time-dependent recoil, bending stiffness and radial strength. The Absorb GT1 (Abbott Vascular, Temecula, CA), DESolve (Elixir Medical Corporation, Sunnyvale, CA) and the Magmaris (BIOTRONIK AG, Bülach, Switzerland) that was previously tested in the BIOSOLVE II study, were tested. Results: Crimped profiles were 1.38 ± 0.01 mm (Absorb GT1), 1.39 ± 0.01 mm (DESolve) and 1.44 ± 0.00 mm (Magmaris) enabling 6F compatibility. Trackability was measured depending on stiffness and force transmission (pushability). Acute elastic recoil was measured at free expansion and within a mock vessel, respectively, yielding results of 5.86 ± 0.76 and 5.22 ± 0.38% (Absorb), 7.85 ± 3.45 and 9.42 ± 0.21% (DESolve) and 5.57 ± 0.72 and 4.94 ± 0.31% (Magmaris). Time-dependent recoil (after 1 h) was observed for the Absorb and DESolve scaffolds but not for the Magmaris. The self-correcting wall apposition behavior of the DESolve did not prevent time-dependent recoil under vessel loading. Conclusions: The results of the suggested test methods allow assessment of technical feasibility based on objective mechanical data and highlight the main differences between polymeric and metallic bioresorbable scaffolds.

  11. In vitro performance investigation of bioresorbable scaffolds – Standard tests for vascular stents and beyond

    International Nuclear Information System (INIS)

    Schmidt, Wolfram; Behrens, Peter; Brandt-Wunderlich, Christoph; Siewert, Stefan; Grabow, Niels; Schmitz, Klaus-Peter

    2016-01-01

    Background/Purpose: Biodegradable polymers are the main materials for coronary scaffolds. Magnesium has been investigated as a potential alternative and was successfully tested in human clinical trials. However, it is still challenging to achieve mechanical parameters comparative to permanent bare metal (BMS) and drug-eluting stents (DES). As such, in vitro tests are required to assess mechanical parameters correlated to the safety and efficacy of the device. Methods/Materials: In vitro bench tests evaluate scaffold profiles, length, deliverability, expansion behavior including acute elastic and time-dependent recoil, bending stiffness and radial strength. The Absorb GT1 (Abbott Vascular, Temecula, CA), DESolve (Elixir Medical Corporation, Sunnyvale, CA) and the Magmaris (BIOTRONIK AG, Bülach, Switzerland) that was previously tested in the BIOSOLVE II study, were tested. Results: Crimped profiles were 1.38 ± 0.01 mm (Absorb GT1), 1.39 ± 0.01 mm (DESolve) and 1.44 ± 0.00 mm (Magmaris) enabling 6F compatibility. Trackability was measured depending on stiffness and force transmission (pushability). Acute elastic recoil was measured at free expansion and within a mock vessel, respectively, yielding results of 5.86 ± 0.76 and 5.22 ± 0.38% (Absorb), 7.85 ± 3.45 and 9.42 ± 0.21% (DESolve) and 5.57 ± 0.72 and 4.94 ± 0.31% (Magmaris). Time-dependent recoil (after 1 h) was observed for the Absorb and DESolve scaffolds but not for the Magmaris. The self-correcting wall apposition behavior of the DESolve did not prevent time-dependent recoil under vessel loading. Conclusions: The results of the suggested test methods allow assessment of technical feasibility based on objective mechanical data and highlight the main differences between polymeric and metallic bioresorbable scaffolds.

  12. Cadmium chloride, benzo[a]pyrene and cyclophosphamide tested in the in vitro mammalian cell micronucleus test (MNvit) in the human lymphoblastoid cell line TK6 at Covance laboratories, Harrogate UK in support of OECD draft Test Guideline 487.

    Science.gov (United States)

    Fowler, Paul; Whitwell, James; Jeffrey, Laura; Young, Jamie; Smith, Katie; Kirkland, David

    2010-10-29

    The following genotoxic chemicals were tested in the in vitro micronucleus assay, at Covance Laboratories, Harrogate, UK in the human lymphoblastoid cell line TK6. Cadmium chloride (an inorganic carcinogen), benzo[a]pyrene (a polycyclic aromatic hydrocarbon requiring metabolic activation) and cyclophosphamide (an alkylating agent requiring metabolic activation) were treated with and without cytokinesis block (by addition of cytochalasin B). This work formed part of a collaborative evaluation of the toxicity measures recommended in the draft OECD Test Guideline 487 for the in vitro micronucleus test. The toxicity measures used, capable of detecting both cytostasis and cell death, were relative population doubling, relative increase in cell counts and relative cell counts for treatments in the absence of cytokinesis block, and replication index or cytokinesis blocked proliferation index in the presence of cytokinesis block. All of the chemicals tested gave significant increases in the percentage of micronucleated cells with and without cytokinesis block at concentrations giving approximately 60% toxicity (cytostasis and cell death) or less by all of the toxicity measures used. The outcomes from this series of tests support the use of relative increase in cell counts and relative population doubling, as well as relative cell counts, as appropriate measures of cytotoxicity for the non-cytokinesis blocked in the in vitro micronucleus assay. Copyright © 2010 Elsevier B.V. All rights reserved.

  13. Ensuring the Quality of Stem Cell-Derived In Vitro Models for Toxicity Testing.

    Science.gov (United States)

    Stacey, Glyn N; Coecke, Sandra; Price, Anna-Bal; Healy, Lyn; Jennings, Paul; Wilmes, Anja; Pinset, Christian; Ingelman-Sundberg, Magnus; Louisse, Jochem; Haupt, Simone; Kidd, Darren; Robitski, Andrea; Jahnke, Heinz-Georg; Lemaitre, Gilles; Myatt, Glenn

    Quality control of cell cultures used in new in vitro toxicology assays is crucial to the provision of reliable, reproducible and accurate toxicity data on new drugs or constituents of new consumer products. This chapter explores the key scientific and ethical criteria that must be addressed at the earliest stages of developing toxicology assays based on human pluripotent stem cell (hPSC) lines. It also identifies key considerations for such assays to be acceptable for regulatory, laboratory safety and commercial purposes. Also addressed is the development of hPSC-based assays for the tissue and cell types of greatest interest in drug toxicology. The chapter draws on a range of expert opinion within the European Commission/Cosmetics Europe-funded alternative testing cluster SEURAT-1 and consensus from international groups delivering this guidance such as the International Stem Cell Banking Initiative. Accordingly, the chapter summarizes the most up-date best practices in the use and quality control of human Pluripotent Stem Cell lines in the development of in vitro toxicity assays from leading experts in the field.

  14. Experiences of the REACH testing proposals system to reduce animal testing.

    Science.gov (United States)

    Taylor, Katy; Stengel, Wolfgang; Casalegno, Carlotta; Andrew, David

    2014-01-01

    In order to reduce animal testing, companies registering chemical substances under the EU REACH legislation must propose rather than conduct certain tests on animals. Third parties can submit 'scientifically valid information' relevant to these proposals to the Agency responsible, the European Chemicals Agency (ECHA), who are obliged to take the information into account. The European Coalition to End Animal Experiments (ECEAE) provided comments on nearly half of the 817 proposals for vertebrate tests on 480 substances published for comment for the first REACH deadline (between 1 August 2009 and 31 July 2012). The paper summarises the response by registrants and the Agency to third party comments and highlights issues with the use of read across, in vitro tests, QSAR and weight of evidence approaches. Use of existing data and evidence that testing is legally or scientifically unjustified remain the most successful comments for third parties to submit. There is a worrying conservatism within the Agency regarding the acceptance of alternative approaches and examples of where registrants have also failed to maximise opportunities to avoid testing.

  15. Three-dimensional in vitro cancer models: a short review

    International Nuclear Information System (INIS)

    Wang, Chengyang; Sun, Wei; Tang, Zhenyu; Li, Lingsong; Zhao, Yu; Yao, Rui

    2014-01-01

    The re-creation of the tumor microenvironment including tumor–stromal interactions, cell–cell adhesion and cellular signaling is essential in cancer-related studies. Traditional two-dimensional (2D) cell culture and animal models have been proven to be valid in some areas of explaining cancerous cell behavior and interpreting hypotheses of possible mechanisms. However, a well-defined three-dimensional (3D) in vitro cancer model, which mimics tumor structures found in vivo and allows cell–cell and cell–matrix interactions, has gained strong interest for a wide variety of diagnostic and therapeutic applications. This communication attempts to provide a representative overview of applying 3D in vitro biological model systems for cancer related studies. The review compares and comments on the differences in using 2D models, animal models and 3D in vitro models for cancer research. Recent technologies to construct and develop 3D in vitro cancer models are summarized in aspects of modeling design, fabrication technique and potential application to biology, pathogenesis study and drug testing. With the help of advanced engineering techniques, the development of a novel complex 3D in vitro cancer model system will provide a better opportunity to understand crucial cancer mechanisms and to develop new clinical therapies. (topical review)

  16. In vitro acute and developmental neurotoxicity screening: an overview of cellular platforms and high-throughput technical possibilities.

    Science.gov (United States)

    Schmidt, Béla Z; Lehmann, Martin; Gutbier, Simon; Nembo, Erastus; Noel, Sabrina; Smirnova, Lena; Forsby, Anna; Hescheler, Jürgen; Avci, Hasan X; Hartung, Thomas; Leist, Marcel; Kobolák, Julianna; Dinnyés, András

    2017-01-01

    Neurotoxicity and developmental neurotoxicity are important issues of chemical hazard assessment. Since the interpretation of animal data and their extrapolation to man is challenging, and the amount of substances with information gaps exceeds present animal testing capacities, there is a big demand for in vitro tests to provide initial information and to prioritize for further evaluation. During the last decade, many in vitro tests emerged. These are based on animal cells, human tumour cell lines, primary cells, immortalized cell lines, embryonic stem cells, or induced pluripotent stem cells. They differ in their read-outs and range from simple viability assays to complex functional endpoints such as neural crest cell migration. Monitoring of toxicological effects on differentiation often requires multiomics approaches, while the acute disturbance of neuronal functions may be analysed by assessing electrophysiological features. Extrapolation from in vitro data to humans requires a deep understanding of the test system biology, of the endpoints used, and of the applicability domains of the tests. Moreover, it is important that these be combined in the right way to assess toxicity. Therefore, knowledge on the advantages and disadvantages of all cellular platforms, endpoints, and analytical methods is essential when establishing in vitro test systems for different aspects of neurotoxicity. The elements of a test, and their evaluation, are discussed here in the context of comprehensive prediction of potential hazardous effects of a compound. We summarize the main cellular characteristics underlying neurotoxicity, present an overview of cellular platforms and read-out combinations assessing distinct parts of acute and developmental neurotoxicology, and highlight especially the use of stem cell-based test systems to close gaps in the available battery of tests.

  17. Toxicity of polymeric nanoparticles in vivo and in vitro

    Science.gov (United States)

    Voigt, Nadine; Henrich-Noack, Petra; Kockentiedt, Sarah; Hintz, Werner; Tomas, Jürgen; Sabel, Bernhard A.

    2014-06-01

    Polybutylcyanoacrylate nanoparticles (PBCA NPs) are candidates for a drug delivery system, which can cross the blood-brain barrier (BBB). Because little is known about their toxicity, we exposed cells to PBCA NPs in vitro and in vivo and monitored their life and death assays. PBCA NPs were fabricated with different surfactants according to the mini-emulsion technique. Viabilities of HeLa and HEK293 cells after NP incubation were quantified by analysing cellular metabolic activity (MTT-test). We then repetitively injected i.v. rhodamine-labelled PBCA NP variations into rats and monitored the survival and morphology of retrogradely labelled neurons by in vivo confocal neuroimaging (ICON) for five weeks. To test for carrier-efficacy and safety, PBCA NPs loaded with Kyotorphin were injected in rats, and a hot plate test was used to quantify analgesic effects. In vitro, we found dose-dependent cell death which was, however, only detectable at very high doses and mainly seen in the cultures incubated with NPs fabricated with the tensids SDS and Tween. However, the in vivo experiments did not show any NP-induced neuronal death, even with particles which were toxic at high dose in vitro, i.e. NPs with Tween and SDS. The increased pain threshold at the hot plate test demonstrated that PBCA NPs are able to cross the BBB and thus comprise a useful tool for drug delivery into the central nervous system (CNS). Our findings showing that different nanoparticle formulations are non-toxic have important implications for the value of NP engineering approaches in medicine.

  18. Impact of bioavailability on the correlation between in vitro cytotoxic and in vivo acute fish toxic concentrations of chemicals

    International Nuclear Information System (INIS)

    Guelden, Michael; Seibert, Hasso

    2005-01-01

    The lower sensitivity of in vitro cytotoxicity assays currently restricts their use as alternative to the fish acute toxicity assays for hazard assessment of chemicals in the aquatic environment. In vitro cytotoxic potencies mostly refer to nominal concentrations. The main objective of the present study was to investigate, whether a reduced availability of chemicals in vitro can account for the lower sensitivity of in vitro toxicity test systems. For this purpose, the bioavailable free fractions of the nominal cytotoxic concentrations (EC 50 ) of chemicals determined with a cytotoxicity test system using Balb/c 3T3 cells and the corresponding free cytotoxic concentrations (ECu 50 ) were calculated. The algorithm applied is based on a previously developed simple equilibrium distribution model for chemicals in cell cultures with serum-supplemented culture media. This model considers the distribution of chemicals between water, lipids and serum albumin. The algorithm requires the relative lipid volume of the test system, the octanol-water partition coefficient (K ow ) and the in vitro albumin-bound fraction of the chemicals. The latter was determined from EC 50 -measurements in the presence of different albumin concentrations with the Balb/c 3T3 test system. Organic chemicals covering a wide range of cytotoxic potency (EC 50 : 0.16-527000 μM) and lipophilicity (log K ow : -5.0-6.96) were selected, for which fish acute toxicity data (LC 50 -values) from at least one of the three fish species, medaka, rainbow trout and fathead minnow, respectively, were available. The availability of several chemicals was shown to be extensively reduced either by partitioning into lipids or by serum albumin binding, or due to both mechanisms. Reduction of bioavailability became more important with increasing cytotoxic potency. The sensitivity of the Balb/c 3T3 cytotoxicity assay and the correspondence between in vivo and in vitro toxic potencies were increased when the free cytotoxic

  19. In vitro detection of cardiotoxins or neurotoxins affecting ion channels or pumps using beating cardiomyocytes as alternative for animal testing

    NARCIS (Netherlands)

    Nicolas, J.A.Y.; Hendriksen, P.J.M.; Haan, de L.H.J.; Koning, R.; Rietjens, I.M.C.M.; Bovee, T.F.H.

    2015-01-01

    The present study investigated if and to what extent murine stem cell-derived beating cardiomyocytes within embryoid bodies can be used as a broad screening in vitro assay for neurotoxicity testing, replacing for example in vivo tests for marine neurotoxins. Effect of nine model compounds, acting on

  20. In vitro bioactivity of glass-ceramic/fibroin composites

    Directory of Open Access Journals (Sweden)

    Lachezar Radev

    2017-06-01

    Full Text Available Bioactive composite materials were prepared by mixing 20 wt.% of silk fibroin (SF and 80 wt.% of glassceramics from CaO-SiO2-P2O5-MgO system. In vitro bioactivity of the prepared composites was evaluated in 1.5 simulated body fluid (1.5 SBF in static conditions. The obtained samples before and after in vitro tests were characterized by X-ray diffraction (XRD analysis, Fourier transform infrared spectroscopy (FTIR, and X-ray photoelectron spectroscopy (XPS. The changes in 1.5 SBF solutions after soaking the samples were evaluated by inductively coupled plasma atomic emission spectroscopy (ICP-AES. MG63 osteosarcoma cells were used for the biological experiments. The obtained experimental data proved that the synthesized composites exhibit excellent in vitro bioactivity.

  1. Evaluation of in vitro and in vivo genotoxicity of single-walled carbon nanotubes.

    Science.gov (United States)

    Kim, Jin Sik; Song, Kyung Seuk; Yu, Il Je

    2015-08-01

    Single-walled carbon nanotubes (SWCNTs) have extensive potential industrial applications due to their unique physical and chemical properties; yet this also increases the chance of human and environment exposure to SWCNTs. Due to the current lack of hazardous effect information on SWNCTs, a standardized genotoxicity battery test was conducted to clarify the genetic toxicity potential of SWCNTs (diameter: 1-1.2 nm, length: ∼20 μm) according to Organization for Economic Cooperation and Development test guidelines 471 (bacterial reverse mutation test), 473 (in vitro chromosome aberration test), and 474 (in vivo micronuclei test) with a good laboratory practice system. The test results showed that the SWCNTs did not induce significant bacterial reverse mutations at 31.3-500 μg/plate in Salmonella typhimurium strains TA98, TA100, TA1535, and TA1537 or in Escherichia coli strain WP2uvrA, with and without a metabolic activation system. Furthermore, the in vitro chromosome aberration test showed no significant increase in structural or numerical chromosome aberration frequencies at SWCNT dose levels of 12.5-50 μg/ml in the presence and absence of metabolic activation. However, dose-dependent cell growth inhibition was found at all the SWCNT dose levels and statistically significant cytotoxic effects observed at certain concentrations in the presence and absence of metabolic activation. Finally, the SWCNTs did not evoke significant in vivo micronuclei frequencies in the polychromatic erythrocytes of an imprinting control region mice at 25-100 mg/kg. Thus, according to the results of the present study, the SWCNTs were not found to have a genotoxic effect on the in vitro and in vivo test systems. © The Author(s) 2013.

  2. Testing Pollen of Single and Stacked Insect-Resistant Bt-Maize on In vitro Reared Honey Bee Larvae

    Science.gov (United States)

    Hendriksma, Harmen P.; Härtel, Stephan; Steffan-Dewenter, Ingolf

    2011-01-01

    The ecologically and economic important honey bee (Apis mellifera) is a key non-target arthropod species in environmental risk assessment (ERA) of genetically modified (GM) crops. Honey bee larvae are directly exposed to transgenic products by the consumption of GM pollen. But most ERA studies only consider responses of adult bees, although Bt-proteins primarily affect the larval phases of target organisms. We adopted an in vitro larvae rearing system, to assess lethal and sublethal effects of Bt-pollen consumption in a standardized eco-toxicological bioassay. The effects of pollen from two Bt-maize cultivars, one expressing a single and the other a total of three Bt-proteins, on the survival and prepupae weight of honey bee larvae were analyzed. The control treatments included pollen from three non-transgenic maize varieties and of Heliconia rostrata. Three days old larvae were fed the realistic exposure dose of 2 mg pollen within the semi-artificial diet. The larvae were monitored over 120 h, until the prepupal stage, where larvae terminate feeding and growing. Neither single nor stacked Bt-maize pollen showed an adverse effect on larval survival and the prepupal weight. In contrast, feeding of H. rostrata pollen caused significant toxic effects. The results of this study indicate that pollen of the tested Bt-varieties does not harm the development of in vitro reared A. mellifera larvae. To sustain the ecosystem service of pollination, Bt-impact on A. mellifera should always be a crucial part of regulatory biosafety assessments. We suggest that our approach of feeding GM pollen on in vitro reared honey bee larvae is well suited of becoming a standard bioassay in regulatory risk assessments schemes of GM crops. PMID:22194811

  3. Testing pollen of single and stacked insect-resistant Bt-maize on in vitro reared honey bee larvae.

    Directory of Open Access Journals (Sweden)

    Harmen P Hendriksma

    Full Text Available The ecologically and economic important honey bee (Apis mellifera is a key non-target arthropod species in environmental risk assessment (ERA of genetically modified (GM crops. Honey bee larvae are directly exposed to transgenic products by the consumption of GM pollen. But most ERA studies only consider responses of adult bees, although Bt-proteins primarily affect the larval phases of target organisms. We adopted an in vitro larvae rearing system, to assess lethal and sublethal effects of Bt-pollen consumption in a standardized eco-toxicological bioassay. The effects of pollen from two Bt-maize cultivars, one expressing a single and the other a total of three Bt-proteins, on the survival and prepupae weight of honey bee larvae were analyzed. The control treatments included pollen from three non-transgenic maize varieties and of Heliconia rostrata. Three days old larvae were fed the realistic exposure dose of 2 mg pollen within the semi-artificial diet. The larvae were monitored over 120 h, until the prepupal stage, where larvae terminate feeding and growing. Neither single nor stacked Bt-maize pollen showed an adverse effect on larval survival and the prepupal weight. In contrast, feeding of H. rostrata pollen caused significant toxic effects. The results of this study indicate that pollen of the tested Bt-varieties does not harm the development of in vitro reared A. mellifera larvae. To sustain the ecosystem service of pollination, Bt-impact on A. mellifera should always be a crucial part of regulatory biosafety assessments. We suggest that our approach of feeding GM pollen on in vitro reared honey bee larvae is well suited of becoming a standard bioassay in regulatory risk assessments schemes of GM crops.

  4. Human Vascular Microphysiological System for in vitro Drug Screening.

    Science.gov (United States)

    Fernandez, C E; Yen, R W; Perez, S M; Bedell, H W; Povsic, T J; Reichert, W M; Truskey, G A

    2016-02-18

    In vitro human tissue engineered human blood vessels (TEBV) that exhibit vasoactivity can be used to test human toxicity of pharmaceutical drug candidates prior to pre-clinical animal studies. TEBVs with 400-800 μM diameters were made by embedding human neonatal dermal fibroblasts or human bone marrow-derived mesenchymal stem cells in dense collagen gel. TEBVs were mechanically strong enough to allow endothelialization and perfusion at physiological shear stresses within 3 hours after fabrication. After 1 week of perfusion, TEBVs exhibited endothelial release of nitric oxide, phenylephrine-induced vasoconstriction, and acetylcholine-induced vasodilation, all of which were maintained up to 5 weeks in culture. Vasodilation was blocked with the addition of the nitric oxide synthase inhibitor L-N(G)-Nitroarginine methyl ester (L-NAME). TEBVs elicited reversible activation to acute inflammatory stimulation by TNF-α which had a transient effect upon acetylcholine-induced relaxation, and exhibited dose-dependent vasodilation in response to caffeine and theophylline. Treatment of TEBVs with 1 μM lovastatin for three days prior to addition of Tumor necrosis factor - α (TNF-α) blocked the injury response and maintained vasodilation. These results indicate the potential to develop a rapidly-producible, endothelialized TEBV for microphysiological systems capable of producing physiological responses to both pharmaceutical and immunological stimuli.

  5. Brain Aggregates: An Effective In Vitro Cell Culture System Modeling Neurodegenerative Diseases.

    Science.gov (United States)

    Ahn, Misol; Kalume, Franck; Pitstick, Rose; Oehler, Abby; Carlson, George; DeArmond, Stephen J

    2016-03-01

    Drug discovery for neurodegenerative diseases is particularly challenging because of the discrepancies in drug effects between in vitro and in vivo studies. These discrepancies occur in part because current cell culture systems used for drug screening have many limitations. First, few cell culture systems accurately model human aging or neurodegenerative diseases. Second, drug efficacy may differ between dividing and stationary cells, the latter resembling nondividing neurons in the CNS. Brain aggregates (BrnAggs) derived from embryonic day 15 gestation mouse embryos may represent neuropathogenic processes in prion disease and reflect in vivo drug efficacy. Here, we report a new method for the production of BrnAggs suitable for drug screening and suggest that BrnAggs can model additional neurological diseases such as tauopathies. We also report a functional assay with BrnAggs by measuring electrophysiological activities. Our data suggest that BrnAggs could serve as an effective in vitro cell culture system for drug discovery for neurodegenerative diseases. © 2016 American Association of Neuropathologists, Inc. All rights reserved.

  6. Evaluation of the medical devices benchmark materials in the controlled human patch testing and in the RhE in vitro skin irritation protocol.

    NARCIS (Netherlands)

    Kandárová, Helena; Bendova, Hana; Letasiova, Silvia; Coleman, Kelly P; De Jong, Wim H; Jírova, Dagmar

    2018-01-01

    Several irritants were used in the in vitro irritation medical device round robin. The objective of this study was to verify their irritation potential using the human patch test (HPT), an in vitro assay, and in vivo data. The irritants were lactic acid (LA), heptanoic acid (HA), sodium dodecyl

  7. Camel milk protein hydrolysates with improved technofunctional properties and enhanced antioxidant potential in in vitro and in food model systems.

    Science.gov (United States)

    Al-Shamsi, Kholoud Awad; Mudgil, Priti; Hassan, Hassan Mohamed; Maqsood, Sajid

    2018-01-01

    Camel milk protein hydrolysates (CMPH) were generated using proteolytic enzymes, such as alcalase, bromelain, and papain, to explore the effect on the technofunctional properties and antioxidant potential under in vitro and in real food model systems. Characterization of the CMPH via degree of hydrolysis, sodium dodecyl sulfate-PAGE, and HPLC revealed that different proteins in camel milk underwent degradation at different degrees after enzymatic hydrolysis using 3 different enzymes for 2, 4, and 6 h, with papain displaying the highest degradation. Technofunctional properties, such as emulsifying activity index, surface hydrophobicity, and protein solubility, were higher in CMPH than unhydrolyzed camel milk proteins. However, the water and fat absorption capacity were lower in CMPH compared with unhydrolyzed camel milk proteins. Antioxidant properties as assessed by 2,2-azinobis(3-ethylbenzthiazoline-6-sulfonic acid) and 2,2-diphenyl-1-picrylhydrazyl radical scavenging activities and metal-chelating activity were enhanced after hydrolysis, in contrast to ferric-reducing antioxidant power which showed a decrease after hydrolysis. The CMPH were also tested in real food model systems for their potential to inhibit lipid peroxidation in fish mince and grape seed oil-in-water emulsion, and we found that papain-produced hydrolysate displayed higher inhibition than alcalase- and bromelain-produced hydrolysates. Therefore, the CMPH demonstrated effective antioxidant potential in vitro as well as in real food systems and showed enhanced functional properties, which guarantees their potential applications in functional foods. The present study is one of few reports available on CMPH being explored in vitro as well as in real food model systems. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  8. Screening of wheat endophytes as biological control agents against Fusarium head blight using two different in vitro tests.

    Science.gov (United States)

    Comby, Morgane; Gacoin, Marie; Robineau, Mathilde; Rabenoelina, Fanja; Ptas, Sébastien; Dupont, Joëlle; Profizi, Camille; Baillieul, Fabienne

    2017-09-01

    In order to find biological control agents (BCAs) for the management of Fusarium head blight (FHB), a major disease on wheat crops worldwide, 86 microorganisms isolated from inner tissues of wheat plants were discriminated for their ability to inhibit the growth of Fusarium graminearum and Fusarium culmorum by in vitro dual culture assays. A group of 22 strains appeared very effective to inhibit F. graminearum (inhibition of 30-51%) and they were also globally effective in controlling F. culmorum (inhibition of 15-53%). Further evaluation of a subselection of strains by screening on detached spikelets in vitro confirmed three species, namely Phoma glomerata, Aureobasidium proteae and Sarocladium kiliense, that have not yet been reported for their efficacy against Fusarium spp., indicating that looking for BCAs toward FHB among wheat endophytes proved to be promising. The efficacy of some strains turned out different between both in vitro screening approaches, raising the importance of finding the most appropriate screening approach for the search of BCAs. This study pointed out the interest of the test on detached wheat spikelets that provided information about a potential pathogenicity, the growth capacity and efficacy of the endophyte strains on the targeted plant, before testing them on whole plants. Copyright © 2017 Elsevier GmbH. All rights reserved.

  9. In Vitro Susceptibility Test of Different Clinical Isolates against Ceftriaxone

    Directory of Open Access Journals (Sweden)

    Syed Hakim Masood

    2010-06-01

    Full Text Available Objectives: Because of the prevailing penicillin resistance in microorganisms, broad spectrum cephalosporins are used empirically specially in developing countries. The aim of this study is to determine the susceptibility pattern of different gram positive and gram negative pathogens against third generation cephalosporin-ceftriaxone to explore the existing effectiveness of this antibiotic.Methods: 180 clinical isolates of different gram positive and gram negative pathogens including P.mirabilis, S. typhi P.aeruginosa, E. coli, S. aureus and Klebsiella were collected from blood and urine samples of in-patients. 30 isolates of all species were tested against each of six brands of ceftriaxone using in vitro sensitivity tests by disc diffusion method (NCCLS criteria. The susceptibility limit was ≥21 mm zone of inhibition, while moderately susceptible was considered at 20-14 mm, and those isolates which showed >13 mm or no zone of inhibition were resistant to this antibacterial drug.Results: Ceftriaxone was found most effective against S. aureus. While 96.1% of the isolates showed susceptibility towards ceftriaxone, followed by E. coli (95%, P. aeruginosa (92.7%, K. pneumonia (89.4% and S. typhi (87.2%. P. mirabilis showed lowest susceptibility amongst all the test organisms (83.8%.Conclusion: Ceftriaxone can be used as a drug of choice in infections caused by S. aureus, E. coli, P. aurigenosa, K. pneumonia and S. typhi. However, it should be used with other antimicrobial agents in order to increase its effectiveness against P. mirabilis.

  10. Error response test system and method using test mask variable

    Science.gov (United States)

    Gender, Thomas K. (Inventor)

    2006-01-01

    An error response test system and method with increased functionality and improved performance is provided. The error response test system provides the ability to inject errors into the application under test to test the error response of the application under test in an automated and efficient manner. The error response system injects errors into the application through a test mask variable. The test mask variable is added to the application under test. During normal operation, the test mask variable is set to allow the application under test to operate normally. During testing, the error response test system can change the test mask variable to introduce an error into the application under test. The error response system can then monitor the application under test to determine whether the application has the correct response to the error.

  11. Implementation challenges for designing Integrated In Vitro Testing Strategies (ITS) aiming at reducing and replacing animal experimentation

    NARCIS (Netherlands)

    Wever, B.de; Fuchs, H.W.; Gaca, M.; Krul, C.A.M.; Mikulowski, S.; Poth, A.; Roggen, E.L.; Vilà, M.R.

    2012-01-01

    At the IVTIP (In Vitro Testing Industrial Platform) meeting of November 26th 2009 entitled 'Toxicology in the 21st century ('21C') - working our way towards a visionary reality' all delegates endorsed the emerging concept of the '21C' vision as the way forward to enable a thorough, reliable and

  12. In vitro cytotoxicity testing of Ubiquicidin 29-41-{sup 99m}Tc

    Energy Technology Data Exchange (ETDEWEB)

    Ocampo, Ivette Z.; Okazaki, Kayo; Dias, Luis Alberto Pereira; Higa, Olga Z.; Silva, Fabiana M. da; Vieira, Daniel P., E-mail: dpvieira@ipen.br [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil); Passos, Priscila; Esteves-Pedro, Natalia M., E-mail: fabiana@biosintesis.com.br [Laboratorio Biosintesis Ltda, Sao Paulo, SP (Brazil)

    2015-07-01

    The work carried out cytotoxicity tests using a radiopharmaceutical compound produced at IPEN/CNEN-SP to certify its safety through in vitro cytotoxicity tests. Since 2009, the Brazilian regulatory agency (ANVISA) requires that such tests have to be carried out following good laboratory practices (GLP) and in according to the OECD (Organisation for Economic Co-operation and Development) guidelines in order to certify its safety for medical use. Those guidelines comprises series of technical recommendations performed to assure quality of experiments. The study chose Ubiquicidin 29-41, an antimicrobial peptide used to discriminate bacterial infection foci from inflammatory sites. Amounts of UBI{sub 29-41} were conjugated or not to {sup 99m}Tc and diluted to equivalent concentrations of 10, 100 and 1000% of the maximum dose (or activity) administered in adults. Possible cytotoxic effects were evaluated in comparison to untreated controls as well as positive and negative damage controls. Both full (radioactive) radiopharmaceuticals, as their precursors (only molecules without conjugation to isotopes) showed no significant cytotoxic effect (cytotoxicity ≤ 10%). The study was conducted for the first time in the country comprising preclinical testing of this radiopharmaceutical in accordance with internationally accepted quality parameters, ensuring the safety of its use and enabling inclusion in the pharmaceutical regulatory agenda. (author)

  13. In-vitro characterization of a cochlear implant system for recording of evoked compound action potentials

    Science.gov (United States)

    2012-01-01

    Background Modern cochlear implants have integrated recording systems for measuring electrically evoked compound action potentials of the auditory nerve. The characterization of such recording systems is important for establishing a reliable basis for the interpretation of signals acquired in vivo. In this study we investigated the characteristics of the recording system integrated into the MED-EL PULSARCI100 cochlear implant, especially its linearity and resolution, in order to develop a mathematical model describing the recording system. Methods In-vitro setup: The cochlear implant, including all attached electrodes, was fixed in a tank of physiologic saline solution. Sinusoidal signals of the same frequency but with different amplitudes were delivered via a signal generator for measuring and recording on a single electrode. Computer simulations: A basic mathematical model including the main elements of the recording system, i.e. amplification and digitalization stage, was developed. For this, digital output for sinusoidal input signals of different amplitudes were calculated using in-vitro recordings as reference. Results Using an averaging of 100 measurements the recording system behaved linearly down to approximately -60 dB of the input signal range. Using the same method, a system resolution of 10 μV was determined for sinusoidal signals. The simulation results were in very good agreement with the results obtained from in-vitro experiments. Conclusions The recording system implemented in the MED-EL PULSARCI100 cochlear implant for measuring the evoked compound action potential of the auditory nerve operates reliably. The developed mathematical model provides a good approximation of the recording system. PMID:22531599

  14. Development of an in vitro skin sensitization test using human cell lines; human Cell Line Activation Test (h-CLAT). II. An inter-laboratory study of the h-CLAT.

    Science.gov (United States)

    Sakaguchi, H; Ashikaga, T; Miyazawa, M; Yoshida, Y; Ito, Y; Yoneyama, K; Hirota, M; Itagaki, H; Toyoda, H; Suzuki, H

    2006-08-01

    Recent regulatory changes have placed a major emphasis on in vitro safety testing and alternative models. In regard to skin sensitization tests, dendritic cells (DCs) derived from human peripheral blood have been considered in the development of new in vitro alternatives. Human cell lines have been also reported recently. In our previous study, we suggested that measuring CD86 and/or CD54 expression on THP-1 cells (human monocytic leukemia cell line) could be used as an in vitro skin sensitization method. An inter-laboratory study among two laboratories was undertaken in Japan in order to further develop an in vitro skin sensitization model. In the present study, we used two human cell lines: THP-1 and U-937 (human histiocytic lymphoma cell line). First we optimized our test protocol (refer to the related paper entitled "optimization of the h-CLAT protocol" within this journal) and then we did an inter-laboratory validation with nine chemicals using the optimized protocol. We measured the expression of CD86 and CD54 on the above cells using flow cytometry after a 24h and 48h exposure to six known allergens (e.g., DNCB, pPD, NiSO(4)) and three non-allergens (e.g., SLS, tween 80). For the sample test concentration, four doses (0.1x, 0.5x, 1x, and 2x of the 50% inhibitory concentration (IC(50))) were evaluated. IC(50) was calculated using MTT assay. We found that allergens/non-allergens were better predicted using THP-1 cells compared to U-937 cells following a 24 h and a 48 h exposure. We also found that the 24h treatment time tended to have a better accuracy than the 48 h treatment time for THP-1 cells. Expression of CD86 and CD54 were good predictive markers for THP-1 cells, but for U-937 cells, expression of CD86 was a better predictor than CD54, at the 24h and the 48 h treatment time. The accuracy also improved when both markers (CD86 and CD54) were used as compared with a single marker for THP-1 cells. Both laboratories gave a good prediction of allergen

  15. In vitro sensitivity of granulo-monocytic progenitors as a new toxicological cell system and endpoint in the ACuteTox Project

    International Nuclear Information System (INIS)

    Cerrato, Laura; Valeri, Antonio; Bueren, Juan A.; Albella, Beatriz

    2009-01-01

    The ACuteTox Project (part of the EU 6th Framework Programme) was started up in January 2005. The aim of this project is to develop a simple and robust in vitro strategy for prediction of human acute systemic toxicity, which could replace animal tests used for regulatory purposes. Our group is responsible for the characterization of the effect of the reference chemicals on the hematopoietic tissue. CFU-GM assay based on the culture of human mononuclear cord blood cells has been used to characterize the effects of the selected compounds on the myeloid progenitors. Previous results have shown the relevance of the CFU-GM assay for the prediction of human acute neutropenia after treatment of antitumoral compounds, and this assay has been recently approved by the ECVAM's Scientific Advisory Committee. Among the compounds included in the study there were pharmaceuticals, environmental pollutants and industrial chemicals. Eleven out of 55 chemicals did not show any cytotoxic effect at the maximum concentration tested. The correlation coefficients of CFU-GM IC50, IC70 and IC90 values with human LC50 values (50% lethal concentration calculated from time-related sublethal and lethal human blood concentrations) were 0.4965, 0.5106 and 0.5142 respectively. Although this correlation is not improve respect to classical in vitro basal cytotoxicity tests such as 3T3 Neutral Red Uptake, chemicals which deviate substantially in the correlation with these assays (colchicine, digoxin, 5-Fluorouracil and thallium sulfate) fitted very well in the linear regression analysis of the CFU-GM progenitors. The results shown in the present study indicate that the sensitivity of CFU-GM progenitors correlates better than the sensitivity of HL-60 cells with human LC50 values and could help to refine the predictability for human acute systemic toxicity when a given chemical may affect to the hematopoietic myeloid system.

  16. A comparative evaluation of in vitro skin sensitisation tests: the human cell-line activation test (h-CLAT) versus the local lymph node assay (LLNA).

    Science.gov (United States)

    Ashikaga, Takao; Sakaguchi, Hitoshi; Sono, Sakiko; Kosaka, Nanae; Ishikawa, Makie; Nukada, Yuko; Miyazawa, Masaaki; Ito, Yuichi; Nishiyama, Naohiro; Itagaki, Hiroshi

    2010-08-01

    We previously developed the human cell-line activation test (h-CLAT) in vitro skin sensitisation test, based on our reported finding that a 24-hour exposure of THP-1 cells (a human monocytic leukaemia cell line) to sensitisers is sufficient to induce the augmented expression of CD86 and CD54. The aim of this study is to confirm the predictive value of h-CLAT for skin sensitisation activity by employing a larger number of test chemicals. One hundred chemicals were selected, according to their categorisation in the local lymph node assay (LLNA), as being: extreme, strong, moderate and weak sensitisers, and non-sensitisers. The correlation of the h-CLAT results with the LLNA results was 84%. There were some false negatives (e.g. benzoyl peroxide, hexyl cinnamic aldehyde) and some false positives (e.g. 1-bromobutane, diethylphthalate). Eight out of the 9 false negatives (89%) were water-insoluble chemicals. The h-CLAT could positively predict not only extreme and strong sensitisers, but also moderate and weak sensitisers, though the detection rates of weak sensitisers and non-sensitisers were comparatively low. Some sensitisers enhanced both CD86 and CD54 levels, and some enhanced the level of only one of them. The use of the combination of CD86 and CD54 induction as a positive indicator, improved the accuracy of the test. In conclusion, the h-CLAT is expected to be a useful cell-based in vitro method for predicting skin sensitisation potential. 2010 FRAME.

  17. [Studies on preparation and dissolution test in vitro of sustained-release dropping pills of curcumin].

    Science.gov (United States)

    Fang, Yu; Xiang, Bai; Pan, Zhen-Hua; Cao, De-Ying

    2010-01-01

    To study the prescription and technique of sustained-release dropping pills of curcumin and inspect their release property in vitro. The orthogonal test was used to screen the prescription and technique which were definited with the colligation evaluation of release and formation of dropping pills. The optimization of prescription and technique were as follows: stearic acid 70 mg, glycery monostearate 25 mg, solutol 6 mg, viscosity of cooling liquid was 100 mm2/s; the temperature of material liquid was 80 degrees C; the cooling temperature was 30 - 0 degrees C; the dropping speed was (21 +/- 2) dripping/min. The release behavior of sustained-release dropping pills of curcumin coincidented with Higuchi equation well and the character of sustained-release was transparent. The sustained-release dropping pills of curcumin have good property of sustained-release in vitro and their release behavior in vivo need to be inspected.

  18. Determination of the potency of a novel saw palmetto supercritical CO2 extract (SPSE for 5α-reductase isoform II inhibition using a cell-free in vitro test system

    Directory of Open Access Journals (Sweden)

    Pais P

    2016-04-01

    Full Text Available Pilar Pais, Agustí Villar, Santiago Rull Euromed, Barcelona, Spain Background: The nicotinamide adenine dinucleotide phosphate-dependent membrane protein 5α-reductase catalyses the conversion of testosterone to the most potent androgen – 5α-dihydrotestosterone. Two 5α-reductase isoenzymes are expressed in humans: type I and type II. The latter is found primarily in prostate tissue. Saw palmetto extract (SPE has been used extensively in the treatment of lower urinary tract symptoms secondary to benign prostatic hyperplasia (BPH. The pharmacological effects of SPE include the inhibition of 5α-reductase, as well as anti-inflammatory and antiproliferative effects. Clinical studies of SPE have been inconclusive – some have shown significant results, and others have not – possibly the result of varying bioactivities of the SPEs used in the studies. Purpose: To determine the in vitro potency in a cell-free test system of a novel SP supercritical CO2 extract (SPSE, an inhibitor of the 5α-reductase isoenzyme type II. Materials and methods: The inhibitory potency of SPSE was compared to that of finasteride, an approved 5α-reductase inhibitor, on the basis of the enzymatic conversion of the substrate androstenedione to the 5α-reduced product 5α-androstanedione. Results: By concentration-dependent inhibition of 5α-reductase type II in vitro (half-maximal inhibitory concentration 3.58±0.05 µg/mL, SPSE demonstrated competitive binding toward the active site of the enzyme. Finasteride, the approved 5α-reductase inhibitor tested as positive control, led to 63%–75% inhibition of 5α-reductase type II. Conclusion: SPSE effectively inhibits the enzyme that has been linked to BPH, and the amount of extract required for activity is comparatively low. It can be confirmed from the results of this study that SPSE has bioactivity that promotes prostate health at a level that is superior to that of many other phytotherapeutic extracts. The

  19. Safety evaluation of food contact paper and board using chemical tests and in vitro bioassays: role of known and unknown substances.

    Science.gov (United States)

    Honkalampi-Hämäläinen, U; Bradley, E L; Castle, L; Severin, I; Dahbi, L; Dahlman, O; Lhuguenot, J-C; Andersson, M A; Hakulinen, P; Hoornstra, D; Mäki-Paakkanen, J; Salkinoja-Salonen, M; Turco, L; Stammati, A; Zucco, F; Weber, A; von Wright, A

    2010-03-01

    In vitro toxicological tests have been proposed as an approach to complement the chemical safety assessment of food contact materials, particularly those with a complex or unknown chemical composition such as paper and board. Among the concerns raised regarding the applicability of in vitro tests are the effects of interference of the extractables on the outcome of the cytotoxicity and genotoxicity tests applied and the role of known compounds present in chemically complex materials, such as paper and board, either as constituents or contaminants. To answer these questions, a series of experiments were performed to assess the role of natural substances (wood extracts, resin acids), some additives (diisopropylnaphthalene, phthalates, acrylamide, fluorescent whitening agents) and contaminants (2,4-diaminotoluene, benzo[a]pyrene) in the toxicological profile of paper and board. These substances were individually tested or used to spike actual paper and board extracts. The toxic concentrations of diisopropylnaphthalenes and phthalates were compared with those actually detected in paper and board extracts showing conspicuous toxicity. According to the results of the spiking experiments, the extracts did not affect the toxicity of tested chemicals nor was there any significant metabolic interference in the cases where two compounds were used in tests involving xenobiotic metabolism by the target cells. While the identified substances apparently have a role in the cytotoxicity of some of the project samples, their presence does not explain the total toxicological profile of the extracts. In conclusion, in vitro toxicological testing can have a role in the safety assessment of chemically complex materials in detecting potentially harmful activities not predictable by chemical analysis alone.

  20. Data Link Test and Analysis System/ATCRBS Transponder Test System Technical Reference

    Science.gov (United States)

    1990-05-01

    This document references material for personnel using or making software changes : to the Data Link Test and Analysis System (DATAS) for Air Traffic Control Radar : Beacon System (ATCRBS) transponder testing and data collection. This is one of : a se...

  1. Inspection system performance test procedure

    International Nuclear Information System (INIS)

    Jensen, C.E.

    1995-01-01

    This procedure establishes requirements to administer a performance demonstration test. The test is to demonstrate that the double-shell tank inspection system (DSTIS) supplied by the contractor performs in accordance with the WHC-S-4108, Double-Shell Tank Ultrasonic Inspection Performance Specification, Rev. 2-A, January, 1995. The inspection system is intended to provide ultrasonic (UT) and visual data to determine integrity of the Westinghouse Hanford Company (WHC) site underground waste tanks. The robotic inspection system consists of the following major sub-systems (modules) and components: Mobile control center; Deployment module; Cable management assembly; Robot mechanism; Ultrasonic testing system; Visual testing system; Pneumatic system; Electrical system; and Control system

  2. Light-Emitting Diode-Based Illumination System for In Vitro Photodynamic Therapy

    OpenAIRE

    Defu Chen; Huifen Zheng; Zhiyong Huang; Huiyun Lin; Zhidong Ke; Shusen Xie; Buhong Li

    2012-01-01

    The aim of this study is to develop a light-emitting diode- (LED-) based illumination system that can be used as an alternative light source for in vitro photodynamic therapy (PDT). This illumination system includes a red LED array composed of 70 LEDs centered at 643 nm, an air-cooling unit, and a specific-designed case. The irradiance as a function of the irradiation distance between the LED array and the sample, the homogeneity and stability of irradiation, and the effect of long-time irrad...

  3. Influence of lipid composition and drug load on the in vitro performance of self-nanoemulsifying drug delivery systems

    DEFF Research Database (Denmark)

    Thomas, Nicky; Müllertz, Anette; Graf, Anja

    2012-01-01

    The influence of lipid composition and drug load on the in vitro performance of lipid-based drug delivery systems was investigated during dispersion and in vitro lipolysis of two self-nanoemulsifying drug delivery systems (SNEDDS). SNEDDS preconcentrates consisted of the same mass ratios of lipid...... of SNEDDS. © 2012 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci 101:1721–1731, 2012...

  4. Toxicity assessment of chemical contaminants;transition from in vitromethods to novel in vitro methods

    Directory of Open Access Journals (Sweden)

    A.A. Farshad

    2007-04-01

    Full Text Available Exposure to occupational and environmental contaminants is a major contributor to human health problems. Despite significant achievements in the risk assessment of chemicals, the toxicological database, particularly for industrial chemicals, remains limited. Considering there areapproximately 80, 000 chemicals in commerce, and an extremely large number of chemical mixtures, in vivo testing of this large number is unachievable from ethical, economical and scientific perspectives. Therefore, increasing the number of available industrial chemicals andnew products has created a demand for alternatives to animal methods for better safety evaluation. Recent toxicity studies have demonstrated that in vitro methods are capable of rapidly providing toxicity information. In this review, current toxicity test methods for risk evaluation of industrial chemical contaminants are presented. To evaluate the potential applications of  more recent test methods developed for toxicity testing of chemical contaminants are discussed. Although  to be considered more broadly for risk assessment of human chemical exposures. In vitro methods,in vitro toxicology methods cannot exactly mimic the biodynamics of the whole body, in vitro  relationships (QSARs and physiologically based toxicokinetic (PBTK models have a potentialtest systems in combination with the knowledge of quantitative structure activity.

  5. Portable Health Algorithms Test System

    Science.gov (United States)

    Melcher, Kevin J.; Wong, Edmond; Fulton, Christopher E.; Sowers, Thomas S.; Maul, William A.

    2010-01-01

    A document discusses the Portable Health Algorithms Test (PHALT) System, which has been designed as a means for evolving the maturity and credibility of algorithms developed to assess the health of aerospace systems. Comprising an integrated hardware-software environment, the PHALT system allows systems health management algorithms to be developed in a graphical programming environment, to be tested and refined using system simulation or test data playback, and to be evaluated in a real-time hardware-in-the-loop mode with a live test article. The integrated hardware and software development environment provides a seamless transition from algorithm development to real-time implementation. The portability of the hardware makes it quick and easy to transport between test facilities. This hard ware/software architecture is flexible enough to support a variety of diagnostic applications and test hardware, and the GUI-based rapid prototyping capability is sufficient to support development execution, and testing of custom diagnostic algorithms. The PHALT operating system supports execution of diagnostic algorithms under real-time constraints. PHALT can perform real-time capture and playback of test rig data with the ability to augment/ modify the data stream (e.g. inject simulated faults). It performs algorithm testing using a variety of data input sources, including real-time data acquisition, test data playback, and system simulations, and also provides system feedback to evaluate closed-loop diagnostic response and mitigation control.

  6. Free Fatty Acid Concentration and Carboxy methyl cellulase Activity of Some Formulas of Protected Fat-proteins Tested In Vitro

    Directory of Open Access Journals (Sweden)

    Lilis Hartati

    2015-05-01

    Full Text Available The aim of this study was to determine the levels of free fatty acids and carboxymethylcellulase activity (cmc-ase activity of some protected fat-proteins base on in vitro Tilley and Terry method. Two sources of fat, i.e. crude palm oil and fish oil and three sources of protein i.e. skim milk, soybean flour and soybean meal were used in the formulation of protected fat-protein, and thus there were six treatment combinations. The filtrate from the in vitro test was analyzed for the levels of free fatty acids and  cmcase activity. The result of this research indicates that different combinations of feed materials and fat give different content of free fatty acid in first stage and second stage in vitro, with the best results in the combination treatment of skim milk and palm oil that give the lowest result of  free fatty acid concentration in fisrt stage in vitro (0.168% and the highest result free fatty acid concentration in second stage in vitro ( 4.312% . The activity of CMC-ase was not influenced by different  sources of fat and protein. It can be concluded was that the protection of the combination between skim milk and CPO gives the highest protection results.

  7. Development of in vitro and in vivo rabies virus neutralization assays based on a high-titer pseudovirus system

    Science.gov (United States)

    Nie, Jianhui; Wu, Xiaohong; Ma, Jian; Cao, Shouchun; Huang, Weijin; Liu, Qiang; Li, Xuguang; Li, Yuhua; Wang, Youchun

    2017-01-01

    Pseudoviruses are useful virological tools because of their safety and versatility; however the low titer of these viruses substantially limits their wider applications. We developed a highly efficient pseudovirus production system capable of yielding 100 times more rabies pseudovirus than the traditional method. Employing the high-titer pseudoviruses, we have developed robust in vitro and in vivo neutralization assays for the evaluation of rabies vaccine, which traditionally relies on live-virus based assays. Compared with current rapid fluorescent focus inhibition test (RFFIT), our in vitro pseudovirus-based neutralization assay (PBNA) is much less labor-intensive while demonstrating better reproducibility. Moreover, the in vivo PBNA assay was also found to be superior to the live virus based assay. Following intravenous administration, the pseudovirus effectively infected the mice, with dynamic viral distributions being sequentially observed in spleen, liver and brain. Furthermore, data from in vivo PBNA showed great agreement with those generated from the live virus model but with the experimental time significantly reduced from 2 weeks to 3 days. Taken together, the effective pseudovirus production system facilitated the development of novel PBNA assays which could replace live virus-based traditional assays due to its safety, rapidity, reproducibility and high throughput capacity. PMID:28218278

  8. In-vitro tensile testing machine for vibration study of fresh rabbit Achilles tendon

    Science.gov (United States)

    Revel, Gian M.; Scalise, Alessandro; Scalise, Lorenzo; Pianosi, Antonella

    2001-10-01

    A lot of people, overall athletic one suffer from tendinitis or complete rupture of the Achilles tendon. This structure becomes inflamed and damaged mainly from a variety of mechanical forces and sometimes due to metabolic problems, such as diabetes or arthritis. Over the past three decades extensive studies have been performed on the structural and mechanical properties of Achilles tendon trying to explain the constitutive equations to describe and foresee tendon behavior. Among the various mechanical parameters, the vibrational behavior is also of interest. Several investigations are performed in order to study how the Achilles tendon vibrations influence the response of the muscle proprioception and human posture. The present article describes how in vitro tensile experiments can be performed, taking into account the need to simulate physiological condition of Achilles tendon and thus approaching some opened problems in the design of the experimental set-up. A new system for evaluating tendon vibrations by non contact techniques is proposed. Preliminary simple elongation tests are made extracting the main mechanical parameters: stress and strain at different fixed stretches, in order to characterize the tissue. Finally, a vibration study is made at each pretensioned tendon level evaluating the oscillating curves caused by a small hammer.

  9. In-vitro diagnostics of Hymenoptera venom allergy

    NARCIS (Netherlands)

    Rueff, F.; Vos, B.; Przybilla, B.

    In-vitro diagnostics of Hymenoptera venom allergy Patients with a history of anaphylactic sting reactions require an allergological work-up (history, in-vitro tests, and skin tests) to clarify indications on venom immunotherapy and on the type of venom to be used. To demonstrate a venom

  10. 5-Fluorouracil, colchicine, benzo[a]pyrene and cytosine arabinoside tested in the in vitro mammalian cell micronucleus test (MNvit) in Chinese hamster V79 cells at Covance Laboratories, Harrogate, UK in support of OECD draft Test Guideline 487.

    Science.gov (United States)

    Whitwell, James; Fowler, Paul; Allars, Sarah; Jenner, Karen; Lloyd, Melvyn; Wood, Debbie; Smith, Katie; Young, Jamie; Jeffrey, Laura; Kirkland, David

    2010-10-29

    The reference genotoxic agents 5-fluorouracil (a nucleoside analogue, characterised by a steep dose response profile), colchicine (an aneugen that inhibits tubulin polymerisation), benzo[a]pyrene (a polycyclic aromatic hydrocarbon requiring metabolic activation) and cytosine arabinoside (a nucleoside analogue that inhibits the gap-filling step of excision repair) were tested in the in vitro micronucleus assay using the Chinese hamster V79 cell line at Covance Laboratories, Harrogate, UK. All chemicals were treated in the absence and presence of cytokinesis block (via addition of cytochalasin B) with this work forming part of a collaborative evaluation of the toxicity measures recommended in the draft OECD Test Guideline 487 on the In Vitro Mammalian Cell Micronucleus Test (MNvit). The toxicity measures used, detecting a possible combination of both cytostasis and cell death (though not cell death directly), were relative population doubling, relative increase in cell counts and relative cell counts for treatments in the absence of cytokinesis block, and replication index in the presence of cytokinesis block. All of the chemicals tested either gave marked increases in the percentage of micronucleated cells with and without cytokinesis block, or did not induce micronuclei at concentrations giving approximately 50-60% toxicity (cytostasis and cell death) or less by all of the toxicity measures used. The outcome from this series of tests supports the use of relative increase in cell counts and relative population doubling, as well as relative cell counts, as appropriate measures of cytotoxicity for the non-cytokinesis blocked in vitro micronucleus assay. Copyright © 2010 Elsevier B.V. All rights reserved.

  11. The 10 basic requirements for a scientific paper reporting antioxidant, antimutagenic or anticarcinogenic potential of test substances in in vitro experiments and animal studies in vivo

    DEFF Research Database (Denmark)

    Verhagen, H.; Aruoma, O.I.; van Delft, J.H.M.

    2003-01-01

    There is increasing evidence that chemicals/test substances cannot only have adverse effects, but that there are many substances that can (also) have a beneficial effect on health. As this journal regularly publishes papers in this area and has every intention in continuing to do so in the near......, provided they can be justified on scientific grounds. The 10 basic requirements for a scientific paper reporting antioxidant, antimutagenic or anticarcinogenic potential of test substances in in vitro experiments and animal studies in vivo concern the following areas: (1) Hypothesis-driven study design; (2......) The nature of the test substance; (3) Valid and invalid test systems; (4) The selection of dose levels and gender; (5) Reversal of the effects induced by oxidants, carcinogens and mutagens; (6) Route of administration; (7) Number and validity of test variables; (8) Repeatability and reproducibility; (9...

  12. Design and validation of a general purpose robotic testing system for musculoskeletal applications.

    Science.gov (United States)

    Noble, Lawrence D; Colbrunn, Robb W; Lee, Dong-Gil; van den Bogert, Antonie J; Davis, Brian L

    2010-02-01

    Orthopaedic research on in vitro forces applied to bones, tendons, and ligaments during joint loading has been difficult to perform because of limitations with existing robotic simulators in applying full-physiological loading to the joint under investigation in real time. The objectives of the current work are as follows: (1) describe the design of a musculoskeletal simulator developed to support in vitro testing of cadaveric joint systems, (2) provide component and system-level validation results, and (3) demonstrate the simulator's usefulness for specific applications of the foot-ankle complex and knee. The musculoskeletal simulator allows researchers to simulate a variety of loading conditions on cadaver joints via motorized actuators that simulate muscle forces while simultaneously contacting the joint with an external load applied by a specialized robot. Multiple foot and knee studies have been completed at the Cleveland Clinic to demonstrate the simulator's capabilities. Using a variety of general-use components, experiments can be designed to test other musculoskeletal joints as well (e.g., hip, shoulder, facet joints of the spine). The accuracy of the tendon actuators to generate a target force profile during simulated walking was found to be highly variable and dependent on stance position. Repeatability (the ability of the system to generate the same tendon forces when the same experimental conditions are repeated) results showed that repeat forces were within the measurement accuracy of the system. It was determined that synchronization system accuracy was 6.7+/-2.0 ms and was based on timing measurements from the robot and tendon actuators. The positioning error of the robot ranged from 10 microm to 359 microm, depending on measurement condition (e.g., loaded or unloaded, quasistatic or dynamic motion, centralized movements or extremes of travel, maximum value, or root-mean-square, and x-, y- or z-axis motion). Algorithms and methods for controlling

  13. In Vitro Susceptibility Testing of Tedizolid against Isolates of Nocardia.

    Science.gov (United States)

    Brown-Elliott, Barbara A; Wallace, Richard J

    2017-12-01

    There is a paucity of efficacious antimicrobials (especially oral) against clinically relevant species of Nocardia To date, all species of Nocardia have been susceptible to linezolid, the first commercially available oxazolidinone. Tedizolid is a new oxazolidinone with previously reported improved in vitro and in vivo (intracellular) potency against multidrug-resistant strains of Mycobacterium sp. and Nocardia brasiliensis Using the current Clinical and Laboratory Standards Institute-recommended broth microdilution method, 101 isolates of Nocardia spp., including 29 Nocardia cyriacigeorgica , 17 Nocardia farcinica , 13 Nocardia nova complex, 21 Nocardia brasiliensis , 5 Nocardia pseudobrasiliensis , and 5 Nocardia wallacei isolates and 11 isolates of less common species, were tested for susceptibility to tedizolid and linezolid. For the most common clinically significant species of Nocardia , tedizolid MIC 50 values were 0.25 μg/ml for N. nova complex, N. brasiliensis , N. pseudobrasiliensis , and N. wallacei , compared to linezolid MIC 50 values of 1, 2, 0.5, and 1 μg/ml, respectively. Tedizolid and linezolid MIC 90 values were 2 μg/ml for N. nova complex and N. brasiliensis Tedizolid MIC 50 and MIC 90 values for both N. cyriacigeorgica and N. farcinica were 0.5 μg/ml and 1 μg/ml, respectively, compared to linezolid MIC 50 and MIC 90 values of 2 and 4 μg/ml, respectively. Based on MIC 90 values, this study showed that tedizolid was 2- to 3-fold more active than linezolid in vitro against most common species of Nocardia , with the exception of the N. nova complex and N. brasiliensis , for which values were the same. These results may warrant evaluation of tedizolid as a potential treatment option for Nocardia infections. Copyright © 2017 American Society for Microbiology.

  14. Changes in the insulin-like growth factor-system may contribute to in vitro age-related impaired osteoblast functions

    DEFF Research Database (Denmark)

    Kveiborg, M.; Rattan, Suresh; Clark, B.F.C.

    2000-01-01

    Age-related bone loss is thought to be due to impaired osteoblast functions. Insulin-like growth factors (IGFs) have been shown to be important stimulators of bone formation and osteoblast activities in vitro and in vivo. We tested the hypothesis that in vitro osteoblast senescence is associated ...

  15. In vitro antifungal susceptibility testing of Scopulariopsis brevicaulis strains using agar diffusion method.

    Science.gov (United States)

    Skóra, Magdalena; Macura, Anna B

    2011-01-01

    The genus Scopulariopsis is a common soil saprotroph and has been isolated from air, organic waste and also from plant, animal and human tissues. Scopulariopsis has mainly been associated in humans with superficial mycoses, but it has also been described as the cause of subcutaneous and invasive infections. The most common aetiological agent of infections in humans is Scopulariopsis brevicaulis. This species has been reported to be resistant in vitro to broad-spectrum antifungal agents available today. The aim of the study was to establish in vitro antifungal susceptibility of 35 S. brevicaulis strains against amphotericin B (AMB), flucytosine (FC), caspofungin (CAS), terbinafine (TER), ciclopirox (CIC), voriconazole (VOR), clotrimazole (CTR), miconazole (MCZ), econazole (ECO), ketoconazole (KET), itraconazole (ITR), and fluconazole (FLU). Antifungal susceptibility tests were evaluated by an agar diffusion method (Neo-Sensitabs, Rosco, Denmark). AMB, FC, CAS, ITR and FLU showed no antifungal activity against S. brevicaulis. TER, CIC, CTR, KET, VOR, ECO, and MCZ revealed inhibitory activity for S. brevicaulis, but it varied for each of the drugs. The best antifungal effect was observed for TER and CIC. All isolates had large inhibition zones for TER and CIC. CTR was also inhibitory for all tested S. brevicaulis isolates, but the diameters of inhibition zones were smaller than for TER and CIC. Nearly 89% isolates showed inhibition zones for KET and the mean diameter of the inhibition zone was comparable to CTR. The least antifungal activity exhibited VQR, ECO and MCZ. Because of the multiresistance of S. brevicaulis, infections due to this species may not respond to particular antifungal treatment and other therapeutic approaches should be considered, e.g., combined therapy and/or surgery.

  16. Implementation challenges for designing integrated in vitro testing strategies (ITS) aiming at reducing and replacing animal experimentation.

    Science.gov (United States)

    De Wever, Bart; Fuchs, Horst W; Gaca, Marianna; Krul, Cyrille; Mikulowski, Stan; Poth, Albrecht; Roggen, Erwin L; Vilà, Maya R

    2012-04-01

    At the IVTIP (in vitro testing industrial platform) meeting of November 26th 2009 entitled 'Toxicology in the 21st century ('21C')--working our way towards a visionary reality' all delegates endorsed the emerging concept of the '21C' vision as the way forward to enable a thorough, reliable and systematic approach to future toxicity testing without the use of animals. One of the emerging concepts focused on integrating a defined number of tests modelling in vivo-relevant and well-characterised toxicity pathways representing mechanistic endpoints. At this meeting the importance of Integrated Testing Strategies (ITS) as tools towards reduction and eventually replacement of the animals currently used for hazard identification and risk assessment was recognised. A follow-up IVTIP Spring 2010 meeting entitled 'Integrated In Vitro Testing Strategies (ITS)--Implementation Challenges' was organised to address pending questions about ITS. This report is not a review of the ITS literature, but a summary of the discussions triggered by presented examples on how to develop and implement ITS. Contrasts between pharmaceutical and chemical industry, as well as a list of general but practical aspects to be considered while developing an ITS emerged from the discussions. In addition, current recommendations on the validation of ITS were discussed. In conclusion, the outcome of this workshop improved the understanding of the participants of some important factors that may impact the design of an ITS in function of its purpose (e.g., screening, or early decision making versus regulatory), the context in which they need to be applied (e.g., ICH guidelines, REACH) and the status and quality of the available tools. A set of recommendations of best practices was established and the importance of the applicability of the individual tests as well as the testing strategy itself was highlighted. Copyright © 2012 Elsevier Ltd. All rights reserved.

  17. An in vitro prototype of a porcine biomimetic testis-like cell culture system: a novel tool for the study of reassembled Sertoli and Leydig cells

    Directory of Open Access Journals (Sweden)

    Iva Arato

    2018-01-01

    Full Text Available At present, there is no reliable in vitro assembled prepubertal testis-like biomimetic organ culture system designed to assess the functional effects of human gonadotropins on Sertoli and Leydig cells. Spermatogenesis is regulated by endocrine, paracrine, and juxtacrine factors (testicular cross-talk, mainly orchestrated by gonadotropins such as luteinizing hormone (LH and follicle-stimulating hormone (FSH that play a pivotal role by stimulating Leydig and Sertoli cells, respectively. The aim of our study was to set up an in vitro prepubertal porcine bioengineered construct as a new model for experimental studies on reassembled Sertoli and Leydig cells. We have evaluated Sertoli and Leydig cells obtained from 15- to 20-day-old neonatal pig testes in terms of purity and function. Subsequently, purified Sertoli and enriched Leydig cells were subjected to coincubation to obtain an in vitro prepubertal porcine testis-like culture system. We performed enzyme-linked immunosorbent assay (ELISA for anti-Müllerian hormone (AMH, inhibin B, and testosterone secretion in the medium, and Real-Time PCR analysis of AMH, inhibin B, FSH-r, aromatase, LHr, and 3β-HSD mRNA expression levels. This in vitro testis-like system was highly responsive to the effects of human gonadotropins and testosterone. AMH mRNA expression and secretion declined, and inhibin-B increased, while FSH-receptor expression was downregulated upon FSH/LH exposure/treatment. Finally, the production of testosterone was increased selectively upon LH treatment. In summary, our proposed model could help to better determine the action of human gonadotropins on Sertoli and Leydig cells. The potential usefulness of the system for shedding light into male infertility-related issues is evident.

  18. In-vitro diagnosis

    DEFF Research Database (Denmark)

    Poulsen, Lars K.

    2001-01-01

    This review on in-vitro diagnostic methods focuses on the use of methods to perform risk assessment on foods. Based on the International Life Science Institute (ILSI) risk decision tree, the methods are discussed and three scenarios are suggested: (i) testing for a well-known allergen; (ii) testing...

  19. IN VITRO SCREENING OF DEVELOPMENTAL NEUROTOXICANTS IN RAT PRIMARY CORTICAL NEURONS USING HIGH CONTENT IMAGE

    Science.gov (United States)

    There is a need for more efficient and cost-effective methods for identifying, characterizing and prioritizing chemicals which may result in developmental neurotoxicity. One approach is to utilize in vitro test systems which recapitulate the critical processes of nervous system d...

  20. In vitro mechanical evaluation of a novel pin-sleeve system for external fixation of distal limb fractures in horses: a proof of concept study.

    Science.gov (United States)

    Brianza, Stefano; Brighenti, Vittoria; Boure, Ludovic; Sprenger, Victor; Pearce, Simon; Schwieger, Karsten

    2010-07-01

    To evaluate the efficacy of a novel pin-sleeve cast (PSC) system for external fixation of distal limb fractures in horses and to compare it with the transfixation pin cast (TPC) system. Experimental. One bone substitute each was used for the TPC and PSC systems. The PSC was tested in 4 configurations characterized by different pin preloads. Specimens were loaded in axial compression in the elastic range. Variables compared statistically were: bone substitute axial displacement and axial strain measured above implants with strain gauges. Pin preload was correlated with the variables investigated. Load to failure and a fatigue tests supplemented the investigation. The PSC configuration with the highest pin preload showed a significantly lower axial displacement compared with the TPC. No significant differences were observed between all other PSC configurations and the TPC. All PSC systems had a significant decrease in recorded strain compared with the TPC system. Pin axial preload inversely correlated with axial displacement but had no effect on axial strain. In the failure test, the PSC encountered plastic deformation earlier than the TPC. In the fatigue test, the PSC ran >200,000 cycles. Preliminary in vitro tests showed that the PSC system significantly reduced peri-implant strain while concurrently having comparable axial displacement to the TPC system. The PSC system has the potential to reduce the risk of pin loosening in horses.

  1. Peri-Implant Strain in an In Vitro Model.

    Science.gov (United States)

    Hussaini, Souheil; Vaidyanathan, Tritala K; Wadkar, Abhinav P; Quran, Firas A Al; Ehrenberg, David; Weiner, Saul

    2015-10-01

    An in vitro experimental model was designed and tested to determine the influence that peri-implant strain may have on the overall crestal bone. Strain gages were attached to polymethylmethacrylate (PMMA) models containing a screw-type root form implant at sites 1 mm from the resin-implant interface. Three different types of crown superstructures (cemented, 1-screw [UCLA] and 2-screw abutment types) were tested. Loading (1 Hz, 200 N load) was performed using a MTS Mechanical Test System. The strain gage data were stored and organized in a computer for statistical treatment. Strains for all abutment types did not exceed the physiological range for modeling and remodeling of cancellous bone, 200-2500 με (microstrain). For approximately one-quarter of the trials, the strain values were less than 200 με the zone for bone atrophy. The mean microstrain obtained was 517.7 με. In conclusion, the peri-implant strain in this in vitro model did not exceed the physiologic range of bone remodeling under axial occlusal loading.

  2. Energy Systems Test Area (ESTA) Electrical Power Systems Test Operations: User Test Planning Guide

    Science.gov (United States)

    Salinas, Michael J.

    2012-01-01

    Test process, milestones and inputs are unknowns to first-time users of the ESTA Electrical Power Systems Test Laboratory. The User Test Planning Guide aids in establishing expectations for both NASA and non-NASA facility customers. The potential audience for this guide includes both internal and commercial spaceflight hardware/software developers. It is intended to assist their test engineering personnel in test planning and execution. Material covered includes a roadmap of the test process, roles and responsibilities of facility and user, major milestones, facility capabilities, and inputs required by the facility. Samples of deliverables, test article interfaces, and inputs necessary to define test scope, cost, and schedule are included as an appendix to the guide.

  3. Validation of capillary blood analysis and capillary testing mode on the epoc Point of Care system.

    Science.gov (United States)

    Cao, Jing; Edwards, Rachel; Chairez, Janette; Devaraj, Sridevi

    2017-12-01

    Laboratory test in transport is a critical component of patient care, and capillary blood is a preferred sample type particularly in children. This study evaluated the performance of capillary blood testing on the epoc Point of Care Blood Analysis System (Alere Inc). Ten fresh venous blood samples was tested on the epoc system under the capillary mode. Correlation with GEM 4000 (Instrumentation Laboratory) was examined for Na+, K+, Cl-, Ca2+, glucose, lactate, hematocrit, hemoglobin, pO2, pCO2, and pH, and correlation with serum tested on Vitros 5600 (Ortho Clinical Diagnostics) was examined for creatinine. Eight paired capillary and venous blood was tested on epoc and ABL800 (Radiometer) for the correlation of Na+, K+, Cl-, Ca2+, glucose, lactate, hematocrit, hemoglobin, pCO2, and pH. Capillary blood from 23 apparently healthy volunteers was tested on the epoc system to assess the concordance to reference ranges used locally. Deming regression correlation coefficients for all the comparisons were above 0.65 except for ionized Ca2+. Accordance of greater than 85% to the local reference ranges were found in all assays with the exception of pO2 and Cl-. Data from this study indicates that capillary blood tests on the epoc system provide comparable results to reference method for these assays, Na+, K+, glucose, lactate, hematocrit, hemoglobin, pCO2, and pH. Further validation in critically ill patients is needed to implement the epoc system in patient transport. This study demonstrated that capillary blood tests on the epoc Point of Care Blood Analysis System give comparable results to other chemistry analyzers for major blood gas and critical tests. The results are informative to institutions where pre-hospital and inter-hospital laboratory testing on capillary blood is a critical component of patient point of care testing.

  4. Results of in vitro chemosensitivity assays

    International Nuclear Information System (INIS)

    Tanigawa, Nobuhiko; Morimoto, Hideki; Akita, Toshiaki; Inoue, Hiroshi; Tanaka, Takeo.

    1986-01-01

    The authors reviewed their experiences to date with chemosensitivity testing of 629 tumors by human tumor clonogenic assay (HTCA) and of 199 tumors by scintillation assay (SA). HTCA and SA were both performed using a double-layer-soft-agar system with continuous exposure of cells to one concentration of standard anticancer drugs. Overall, 60 % of specimens in HTCA and 58 % in SA produced significant growth in vitro. HTCA was 52 % (13/25) reliable for predicting in vivo sensitivity, and 95 % (36/38) reliable for in vivo resistance, whereas SA was 40 % (8/20) reliable for in vivo sensitivity and 88 % (21/24) for in vivo resistance. In vitro success rates were variable, depending on the tumor histology. In vitro growth of gastric cancer specimens was characteristically lower than that of colon cancer specimens (48 % and 60 % in HTCA, and 46 % and 68 % in SA, respectively). (p < 0.005). Optimal in vitro-in vivo drug concentrations and culture conditions are still being defined. Correlation studies of in vitro-in vivo responses of gastrointestinal cancers suggested that in vitro concentrations of 5-fluorouracil and mitomycin C used in this study were considerably higher than their optimal doses. Tumor cell heterogeneity poses significant problems in the clinical use of chemosensitivity assays. In this last study, we sought evidence of tumor heterogeneity by comparing chemosensitivity responses between : 1) different portions of a single tumor, 2) a primary and a metastatic biopsy taken from a patient on the same day, and 3) different metastases from a patient taken on the same day. The results demonstrated the presence of considerable heterogeneity of response to chemotherapy among different tumors from the same patient, and even within the same tumor. The reported discrepancies of in vitro and in vivo sensitivity may be due to such therapeutic heterogeneity among tumors. (J.P.N.)

  5. [In vitro activity of ertapenem against clinical bacterial isolates in 69 Spanish medical centers (E-test study)].

    Science.gov (United States)

    Gobernado, M; Sanz-Rodríguez, C; Villanueva, R; Torroba, L; Redondo, E; González-Esteban, J

    2007-12-01

    This study was conducted to assess the in vitro activity of ertapenem against clinical bacterial isolates from patients with community-acquired intra-abdominal and lower tract respiratory infections in Spain in 2003. As the study was conducted before the marketing of ertapenem, it was also useful to define a baseline susceptibility pattern for ertapenem in each of the participating hospitals for later surveillance studies. Each partipating site identified a variable number of aerobic and facultative bacteria isolated from patients with community-acquired intra-abdominal infection or pneumonia using standard procedures. E-test strips were used for determining the minimum inhibitory concentration (MIC) of ertapenem, while for other antimicrobials either quantitative dilution techniques or qualitative diffusion procedures were used according to each microbiology laboratory's routine practice. MIC breakpoints for categorization of susceptibility provided by the CLSI were used for interpreting MIC values. A total of 2,901 recent clinical isolates from patients with community-acquired intra-abdominal infection or pneumonia hospitalized in 69 Spanish medical centers were tested. These isolates included 2,039 Gram-negative bacteria (1,646 Enterobacteriaceae, 216 Haemophilus, 123 non-fermenting Gram-negative bacteria [NFGNB] and 54 others) and 862 Gram-positive bacteria (556 pneumococci, 159 staphylococci, 96 streptococci other than S. pneumoniae, 44 enterococci and 7 others). Ertapenem was very active in vitro against Enterobacteriaceae (99.8% susceptible), Haemophilus (96.3% susceptible), pneumococci (99.6% susceptible, of which 31% were penicillin non-susceptible strains), streptococci other than S. pneumoniae (99.0% susceptible) and methicillin-susceptible staphylococci (94.8% susceptible). For other Gram-positive and Gram-negative pathogens for which ertapenem susceptible breakpoints have not been defined, MIC(90) values were 0.38 and 0.064 mg/l, respectively. As

  6. In vitro culture of coconut (Cocos nucifera L.) zygotic embryos.

    Science.gov (United States)

    Engelmann, Florent; Malaurie, Bernard; N'Nan, Oulo

    2011-01-01

    Coconut is a very important crop for millions of people in tropical countries. With coconut, in vitro culture protocols have been developed with two main objectives, viz. the large scale production of particular types of coconuts and the international exchange and conservation of coconut germplasm. The methods described in this chapter have been developed in the framework of collaborative activities between research institutes in Côte d'Ivoire and France. Two coconut embryo in vitro collecting protocols have been established, one consisting of storing the disinfected embryos in a KCl solution until they are brought back to the laboratory, where they are re-disinfected and inoculated in vitro under sterile conditions, and the other including in vitro inoculation of the embryos in the field. For international germplasm exchange, zygotic embryos inoculated in vitro in plastic test tubes or endosperm cylinders containing embryos in plastic bags are used. For in vitro culture, embryos are inoculated on semi-solid medium supplemented with sucrose and activated charcoal and placed in the dark, and then transferred to light conditions with the same (solid or liquid) medium once the first true leaf is visible and the root system has started developing.

  7. Energy Systems High-Pressure Test Laboratory | Energy Systems Integration

    Science.gov (United States)

    Facility | NREL Energy Systems High-Pressure Test Laboratory Energy Systems High-Pressure Test Laboratory In the Energy Systems Integration Facility's High-Pressure Test Laboratory, researchers can safely test high-pressure hydrogen components. Photo of researchers running an experiment with a hydrogen fuel

  8. In vitro Fab display: a cell-free system for IgG discovery

    Science.gov (United States)

    Stafford, Ryan L.; Matsumoto, Marissa L.; Yin, Gang; Cai, Qi; Fung, Juan Jose; Stephenson, Heather; Gill, Avinash; You, Monica; Lin, Shwu-Hwa; Wang, Willie D.; Masikat, Mary Rose; Li, Xiaofan; Penta, Kalyani; Steiner, Alex R.; Baliga, Ramesh; Murray, Christopher J.; Thanos, Christopher D.; Hallam, Trevor J.; Sato, Aaron K.

    2014-01-01

    Selection technologies such as ribosome display enable the rapid discovery of novel antibody fragments entirely in vitro. It has been assumed that the open nature of the cell-free reactions used in these technologies limits selections to single-chain protein fragments. We present a simple approach for the selection of multi-chain proteins, such as antibody Fab fragments, using ribosome display. Specifically, we show that a two-chain trastuzumab (Herceptin) Fab domain can be displayed in a format which tethers either the heavy or light chain to the ribosome while retaining functional antigen binding. Then, we constructed synthetic Fab HC and LC libraries and performed test selections against carcinoembryonic antigen (CEA) and vascular endothelial growth factor (VEGF). The Fab selection output was reformatted into full-length immunoglobulin Gs (IgGs) and directly expressed at high levels in an optimized cell-free system for immediate screening, purification and characterization. Several novel IgGs were identified using this cell-free platform that bind to purified CEA, CEA positive cells and VEGF. PMID:24586053

  9. Evolution of a test article handling system for the SP-100 ground engineering system test

    International Nuclear Information System (INIS)

    Shen, E.J.; Schweiger, L.J.; Miller, W.C.; Gluck, R.; Devies, S.M.

    1987-04-01

    A simulated space environment test of a flight prototypic SP-100 reactor, control system, and flight shield will be conducted at the Hanford Engineering Development Laboratory (HEDL). The flight prototypic components and the supporting primary heat removal system are collectively known as the Nuclear Assembly Test Article (TA). The unique configuration and materials of fabrication for the Test Article require a specialized handling facility to support installation, maintenance, and final disposal operations. Westinghouse Hanford Company, the Test Site Operator, working in conjunction with General Electric Company, the Test Article supplier, developed and evaluated several handling concepts resulting in the selection of a reference Test Article Handling System. The development of the reference concept for the handling system is presented

  10. In Vitro Antibiotic Susceptibilities of Burkholderia mallei (Causative Agent of Glanders) Determined by Broth Microdilution and E-Test

    Science.gov (United States)

    Heine, Henry S.; England, Marilyn J.; Waag, David M.; Byrne, W. Russell

    2001-01-01

    In vitro susceptibilities to 28 antibiotics were determined for 11 strains of Burkholderia mallei by the broth microdilution method. The B. mallei strains demonstrated susceptibility to aminoglycosides, macrolides, quinolones, doxycycline, piperacillin, ceftazidime, and imipenem. For comparison and evaluation, 17 antibiotic susceptibilities were also determined by the E-test. E-test values were always lower than the broth dilution values. Establishing and comparing antibiotic susceptibilities of specific B. mallei strains will provide reference information for assessing new antibiotic agents. PMID:11408233

  11. An In Vitro System Comprising Immortalized Hypothalamic Neuronal Cells (GT1-7 Cells) for Evaluation of the Neuroendocrine Effects of Essential Oils.

    Science.gov (United States)

    Mizuno, Dai; Konoha-Mizuno, Keiko; Mori, Miwako; Yamazaki, Kentaro; Haneda, Toshihiro; Koyama, Hironari; Kawahara, Masahiro

    2015-01-01

    Aromatherapy and plant-based essential oils are widely used as complementary and alternative therapies for symptoms including anxiety. Furthermore, it was reportedly effective for the care of several diseases such as Alzheimer's disease and depressive illness. To investigate the pharmacological effects of essential oils, we developed an in vitro assay system using immortalized hypothalamic neuronal cells (GT1-7 cells). In this study, we evaluated the effects of essential oils on neuronal death induced by hydrogen peroxide (H2O2), aluminum, zinc, or the antagonist of estrogen receptor (tamoxifen). Among tests of various essential oils, we found that H2O2-induced neuronal death was attenuated by the essential oils of damask rose, eucalyptus, fennel, geranium, ginger, kabosu, mandarin, myrrh, and neroli. Damask rose oil had protective effects against aluminum-induced neurotoxicity, while geranium and rosemary oil showed protective activity against zinc-induced neurotoxicity. In contrast, geranium oil and ginger oil enhanced the neurotoxicity of tamoxifen. Our in vitro assay system could be useful for the neuropharmacological and endocrine pharmacological studies of essential oils.

  12. Multi-Organ toxicity demonstration in a functional human in vitro system composed of four organs.

    Science.gov (United States)

    Oleaga, Carlota; Bernabini, Catia; Smith, Alec S T; Srinivasan, Balaji; Jackson, Max; McLamb, William; Platt, Vivien; Bridges, Richard; Cai, Yunqing; Santhanam, Navaneetha; Berry, Bonnie; Najjar, Sarah; Akanda, Nesar; Guo, Xiufang; Martin, Candace; Ekman, Gail; Esch, Mandy B; Langer, Jessica; Ouedraogo, Gladys; Cotovio, Jose; Breton, Lionel; Shuler, Michael L; Hickman, James J

    2016-02-03

    We report on a functional human model to evaluate multi-organ toxicity in a 4-organ system under continuous flow conditions in a serum-free defined medium utilizing a pumpless platform for 14 days. Computer simulations of the platform established flow rates and resultant shear stress within accepted ranges. Viability of the system was demonstrated for 14 days as well as functional activity of cardiac, muscle, neuronal and liver modules. The pharmacological relevance of the integrated modules were evaluated for their response at 7 days to 5 drugs with known side effects after a 48 hour drug treatment regime. The results of all drug treatments were in general agreement with published toxicity results from human and animal data. The presented phenotypic culture model exhibits a multi-organ toxicity response, representing the next generation of in vitro systems, and constitutes a step towards an in vitro "human-on-a-chip" assay for systemic toxicity screening.

  13. Comparison of in vitro activity of undecylenic acid and tolnaftate against athlete's foot fungi.

    Science.gov (United States)

    Amsel, L P; Cravitz, L; VanderWyk, R; Zahry, S

    1979-03-01

    Undecylenic acid and tolnaftate were tested in an in vitro test system to evaluate their relative "killing time" efficacy against Trichophyton mentagrophytes, Trichophyton rubrum, and Epidermophyton floccosum. Commercial products containing these active agents were tested similarly. The pure active agents were equivalent in activity. The commercial product containing undecylenic acid appeared to be more effective against the test organisms than did the product containing tolnaftate.

  14. Development of an in vitro test battery for assessing chemical effects on bovine germ cells under the ReProTect umbrella

    International Nuclear Information System (INIS)

    Lazzari, Giovanna; Tessaro, Irene; Crotti, Gabriella; Galli, Cesare; Hoffmann, Sebastian; Bremer, Susanne; Pellizzer, Cristian

    2008-01-01

    Current European legislation for the registration and authorisation of chemicals (REACH) will require a dramatic increase in the use of animals for reproductive toxicity testing. Since one objective of REACH is to use vertebrates only as last resort, the development and validation of alternative methods is urgently needed. For this purpose ReProTect, an integrated research project funded by the European Union, joining together 33 partners with complementary expertise in reproductive toxicology, was designed. The study presented here describes a battery of two tests developed within ReProTect. The objective of these tests is the detection of chemical effects during the processes of oocyte maturation and fertilisation in a bovine model. The corresponding toxicological endpoints are the reaching of metaphase II and the formation of the pronuclei respectively. Fifteen chemicals have been tested (Benzo[a]pyrene, Busulfan, Butylparaben, Cadmium Chloride, Carbendazim, Cycloheximide, Diethylstilbestrol, Genistein, Ionomycin, Ketoconazole, Lindane, Methylacetoacetate, Mifepristone, Nocodazole and DMSO as solvent) demonstrating high intra-laboratory reproducibility of the tests. Furthermore, the responses obtained in both tests, for several substances, had a good correlation with the available in vivo and in vitro data. These tests therefore, could predictably become part of an integrated testing strategy that combines the bovine models with additional in vitro tests, in order to predict chemical hazards on mammalian fertility

  15. Radioactively labelled substances for in vivo and in vitro tests from the time of their production up to their use in nuclear medicine

    International Nuclear Information System (INIS)

    Riccato, M.T.

    1976-01-01

    Most important production steps and control tests applied to radionuclides and radioactive labelled compounds used in nuclear medicine are described. Radiopharmaceutical products are subdivided in products for therapy and for in vivo and in vitro diagnostics tests. (orig./HP) [de

  16. The ability of in vitro antioxidant assays to predict the efficiency of a cod protein hydrolysate and brown seaweed extract to prevent oxidation in marine food model systems.

    Science.gov (United States)

    Jónsdóttir, Rósa; Geirsdóttir, Margrét; Hamaguchi, Patricia Y; Jamnik, Polona; Kristinsson, Hordur G; Undeland, Ingrid

    2016-04-01

    The ability of different in vitro antioxidant assays to predict the efficiency of cod protein hydrolysate (CPH) and Fucus vesiculosus ethyl acetate extract (EA) towards lipid oxidation in haemoglobin-fortified washed cod mince and iron-containing cod liver oil emulsion was evaluated. The progression of oxidation was followed by sensory analysis, lipid hydroperoxides and thiobarbituric acid-reactive substances (TBARS) in both systems, as well as loss of redness and protein carbonyls in the cod system. The in vitro tests revealed high reducing capacity, high DPPH radical scavenging properties and a high oxygen radical absorbance capacity (ORAC) value of the EA which also inhibited lipid and protein oxidation in the cod model system. The CPH had a high metal chelating capacity and was efficient against oxidation in the cod liver oil emulsion. The results indicate that the F. vesiculosus extract has a potential as an excellent natural antioxidant against lipid oxidation in fish muscle foods while protein hydrolysates are more promising for fish oil emulsions. The usefulness of in vitro assays to predict the antioxidative properties of new natural ingredients in foods thus depends on the knowledge about the food systems, particularly the main pro-oxidants present. © 2015 Society of Chemical Industry.

  17. Space Fission System Test Effectiveness

    International Nuclear Information System (INIS)

    Houts, Mike; Schmidt, Glen L.; Van Dyke, Melissa; Godfroy, Tom; Martin, James; Bragg-Sitton, Shannon; Dickens, Ricky; Salvail, Pat; Harper, Roger

    2004-01-01

    Space fission technology has the potential to enable rapid access to any point in the solar system. If fission propulsion systems are to be developed to their full potential, however, near-term customers need to be identified and initial fission systems successfully developed, launched, and utilized. One key to successful utilization is to develop reactor designs that are highly testable. Testable reactor designs have a much higher probability of being successfully converted from paper concepts to working space hardware than do designs which are difficult or impossible to realistically test. ''Test Effectiveness'' is one measure of the ability to realistically test a space reactor system. The objective of this paper is to discuss test effectiveness as applied to the design, development, flight qualification, and acceptance testing of space fission systems. The ability to perform highly effective testing would be particularly important to the success of any near-term mission, such as NASA's Jupiter Icy Moons Orbiter, the first mission under study within NASA's Project Prometheus, the Nuclear Systems Program

  18. An in vitro system from Plasmodium falciparum active in endogenous mRNA translation

    Directory of Open Access Journals (Sweden)

    Ferreras Ana

    2000-01-01

    Full Text Available An in vitro translation system has been prepared from Plasmodium falciparum by saponin lysis of infected-erythrocytes to free parasites which were homogeneized with glass beads, centrifuged to obtain a S-30 fraction followed by Sephadex G-25 gel filtration. This treatment produced a system with very low contamination of host proteins (<1%. The system, optimized for Mg2+ and K+, translates endogenous mRNA and is active for 80 min which suggests that their protein factors and mRNA are quite stable.

  19. Test results of HTTR control system

    International Nuclear Information System (INIS)

    Motegi, Toshihiro; Iigaki, Kazuhiko; Saito, Kenji; Sawahata, Hiroaki; Hirato, Yoji; Kondo, Makoto; Shibutani, Hideki; Ogawa, Satoru; Shinozaki, Masayuki; Mizushima, Toshihiko; Kawasaki, Kozo

    2006-06-01

    The plant control performance of the IHX helium flow rate control system, the PPWC helium flow rate control system, the secondary helium flow rate control system, the inlet temperature control system, the reactor power control system and the outlet temperature control system of the HTTR are obtained through function tests and power-up tests. As the test results, the control systems show stable control response under transient condition. Both of inlet temperature control system and reactor power control system shows stable operation from 30% to 100%, respectively. This report describes the outline of control systems and test results. (author)

  20. Development of an in vitro skin sensitization test based on ROS production in THP-1 cells.

    Science.gov (United States)

    Saito, Kazutoshi; Miyazawa, Masaaki; Nukada, Yuko; Sakaguchi, Hitoshi; Nishiyama, Naohiro

    2013-03-01

    Recently, it has been reported that reactive oxygen species (ROS) produced by contact allergens can affect dendritic cell migration and contact hypersensitivity. The aim of the present study was to develop a new in vitro assay that could predict the skin sensitizing potential of chemicals by measuring ROS production in THP-1 (human monocytic leukemia cell line) cells. THP-1 cells were pre-loaded with a ROS sensitive fluorescent dye, 5-(and 6-)-chloromethyl-2', 7'-dichlorodihydrofluorescein diacetate, acetyl ester (CM-H2DCFDA), for 15min, then incubated with test chemicals for 30min. The fluorescence intensity was measured by flow cytometry. For the skin sensitizers, 25 out of 30 induced over a 2-fold ROS production at more than 90% of cell viability. In contrast, increases were only seen in 4 out of 20 non-sensitizers. The overall accuracy for the local lymph node assay (LLNA) was 82% for 50 chemicals tested. A correlation was found between the estimated concentration showing 2-fold ROS production in the ROS assay and the EC3 values (estimated concentration required to induce positive response) of the LLNA. These results indicated that the THP-1 cell-based ROS assay was a rapid and highly sensitive detection system able to predict skin sensitizing potentials and potency of chemicals. Copyright © 2012 Elsevier Ltd. All rights reserved.

  1. Pharmacological effects of primaquine ureas and semicarbazides on the central nervous system in mice and antimalarial activity in vitro.

    Science.gov (United States)

    Kedzierska, Ewa; Orzelska, Jolanta; Perković, Ivana; Knežević, Danijel; Fidecka, Sylwia; Kaiser, Marcel; Zorc, Branka

    2016-02-01

    New primaquine (PQ) urea and semicarbazide derivatives 1-4 were screened for the first time for central nervous system (CNS) and antimalarial activity. Behavioural tests were performed on mice. In vitro cytotoxicity on L-6 cells and activity against erythrocytic stages of Plasmodium falciparum was determined. Compound 4 inhibited 'head-twitch' responses and decreased body temperature of mice, which suggests some involvement of the serotonergic system. Compound 4 protected mice against clonic seizures and was superior in the antimalarial test. A hybrid of two PQ urea 2 showed a strong antimalarial activity, confirming the previous findings of the high activity of bis(8-aminoquinolines) and other bisantimalarial drugs. All the compounds decreased the locomotor activity of mice, what suggests their weak depressive effects on the CNS, while PQ derivatives 1 and 2 increased amphetamine-induced hyperactivity. None of the compounds impaired coordination, what suggests a lack of their neurotoxicity. All the tested compounds presented an antinociceptive activity in the 'writhing' test. Compounds 3 and 4 were active in nociceptive tests, and those effects were reversed by naloxone. Compound 4 could be a useful lead compound in the development of CNS active agents and antimalarials, whereas compound 3 may be considered as the most promising lead for new antinociceptive agents. © 2015 Société Française de Pharmacologie et de Thérapeutique.

  2. Lipofection of early passages of cell cultures derived from murine adenocarcinomas: in vitro and ex vivo testing of the thymidine kinase/ganciclovir system.

    Science.gov (United States)

    Karara, Armando L; Bumaschny, Viviana F; Fiszman, Gabriel L; Casais, Cecilia C; Glikin, Gerardo C; Finocchiaro, Liliana Me

    2002-01-01

    Early passages of cultured cells derived from four spontaneous Balb/c murine adenocarcinomas were used to explore the feasibility of a nonviral HSVtk-based suicide gene therapy system. After lipofection with pCMVtk, the transiently HSVtk expressing P07 (lung), M3, M05, and M38 (mammary gland) cells were, respectively, about 130-, 30-, 120-, and 170-fold more sensitive to ganciclovir (GCV) in vitro than their respective controls. Eighty percent of Balb/c mice subcutaneously inoculated with ex vivo pCMVtk-lipofected P07 cells, followed by intraperitoneal GCV injection for 7 days, displayed a complete inhibition of tumor growth for over 70 days. Control animals started to display tumors 13 days after inoculation. We present evidence showing that early passages of cultured tumor cells can efficiently express lipofected genes and that they are sensitive to the lipoplex-mediated HSVtk/GCV system.

  3. Application of a new MR Microscope using an Independent Console System (MRMICS) for biological tissues in vitro

    International Nuclear Information System (INIS)

    Yoshioka, Hiroshi; Anno, Izumi; Itai, Yuji; Haishi, Tomoyuki; Adachi, Naotaka; Kose, Katsumi

    1999-01-01

    We studied microscopic MR images of the normal appendix in vitro using a new MR microscope system: MR Microscope using an Independent Console System (MRMICS). The MRMICS was placed in the clinical MR room, and the probe box was fixed on the bed of the 1.5 T clinical MR machine. T1-, T2-, and proton density-weighted images were obtained using spin echo sequences with an in-plane pixel size of 100 x 100 μm. Zonal structures of the appendix were clearly demonstrated with different contrast by different sequences. Therefore, the MRMICS is a useful add-on system for investigating microscopic MR images of biological tissues in vitro. (author)

  4. Application of a new MR Microscope using an Independent Console System (MRMICS) for biological tissues in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Yoshioka, Hiroshi; Anno, Izumi; Itai, Yuji [Tsukuba Univ., Ibaraki (Japan). Inst. of Clinical Medicine; Haishi, Tomoyuki; Adachi, Naotaka; Kose, Katsumi

    1999-02-01

    We studied microscopic MR images of the normal appendix in vitro using a new MR microscope system: MR Microscope using an Independent Console System (MRMICS). The MRMICS was placed in the clinical MR room, and the probe box was fixed on the bed of the 1.5 T clinical MR machine. T1-, T2-, and proton density-weighted images were obtained using spin echo sequences with an in-plane pixel size of 100 x 100 {mu}m. Zonal structures of the appendix were clearly demonstrated with different contrast by different sequences. Therefore, the MRMICS is a useful add-on system for investigating microscopic MR images of biological tissues in vitro. (author)

  5. Evaluation of a three compartment in vitro gastrointestinal simulator dissolution apparatus to predict in vivo dissolution.

    Science.gov (United States)

    Takeuchi, Susumu; Tsume, Yasuhiro; Amidon, Gregory E; Amidon, Gordon L

    2014-11-01

    In vitro dissolution tests are performed for new formulations to evaluate in vivo performance, which is affected by the change of gastrointestinal (GI) physiology, in the GI tract. Thus, those environmental changes should be introduced to an in vitro dissolution test. Many studies have successfully shown the improvement of in vitro-in vivo correlations (IVIVC) by introducing those physiological changes into dissolution tests. The gastrointestinal simulator (GIS), a multicompartment in vitro dissolution apparatus, was developed to evaluate in vivo drug dissolution. A gastric-emptying rate along with transit rate are key factors to evaluate in vivo drug dissolution and, hence, drug absorption. Dissolution tests with the GIS were performed with Biopharmaceutical Classification System class I drugs at five different gastric-emptying rates in the fasted state. Computational models were used to determine in vivo gastric-emptying time for propranolol and metoprolol based on the GIS dissolution results. Those were compared with published clinical data to determine the gastric half-emptying time. In conclusion, the GIS is a practical tool to assess dissolution properties and can improve IVIVC. © 2014 Wiley Periodicals, Inc. and the American Pharmacists Association.

  6. Exploring the fate of liposomes in the intestine by dynamic in vitro lipolysis

    DEFF Research Database (Denmark)

    Parmentier, Johannes; Thomas, Nicky; Müllertz, Anette

    2012-01-01

    precipitation was detected during the lipolysis assay, despite pronounced lipolytic degradation and change in vesicle size. In conclusion, the tested dynamic in vitro lipolysis model is suitable for the assessment of liposome stability in the intestine. Furthermore, liposomes might be a useful alternative......Liposomes are generally well tolerated drug delivery systems with a potential use for the oral route. However, little is known about the fate of liposomes during exposure to the conditions in the gastro-intestinal tract (GIT). To gain a better understanding of liposome stability in the intestine......, a dynamic in vitro lipolysis model, which so far has only been used for the in vitro characterisation of other lipid-based drug delivery systems, was applied to different liposomal formulations. Liposome size and phospholipid (PL) digestion were determined as two markers for liposome stability. In addition...

  7. In vitro systems in pneumocystis research

    DEFF Research Database (Denmark)

    Dei-Cas, E; Cailliez, J C; Lundgren, Bettina

    1996-01-01

    as well as elimination of host cell debris from parasite extracts. In other investigations, such as transmission, infectivity, life cycle, biochemical, in vitro culture or drug-screening studies, viable and infectious Pneumocystis organisms are urgently required. However, there is no generally accepted...

  8. Testing lifting systems in nuclear facilities

    International Nuclear Information System (INIS)

    Kling, H.; Laug, R.

    1984-01-01

    Lifting systems in nuclear facilities must be inspected at regular intervals after having undergone their first acceptance test. These inspections are frequently carried out by service firms which not only employ the skilled personnel required for such jobs but also make available the necessary test equipment. The inspections in particular include a number of sophisticated load tests for which test load systems have been developed to allow lifting systems to be tested so that reactor specific boundary conditions are taken into account. In view of the large number of facilities to be inspected, the test load system is a modular system. (orig.) [de

  9. Quantitative evaluation of apical extrusion of debris and irrigants using four rotary instrumentation systems: an in vitro study.

    Science.gov (United States)

    Nagaveni, S Aspalli; Balakoti, K Reddy; Smita, Karan; Ratnakar, P; Satish, S V; Aravind, T

    2013-11-01

    The apical extrusion of infected debris may have the potential to disrupt the balance between microbial aggression and host defense, resulting in incidents of acute inflammation. During preparation, irrigants and debris, such as bacteria, dentin filings and necrotic tissue may be extruded into the periradicular region leading to periapical inflammation and postoperative flare ups. Using an instrumentation technique that minimizes apical extrusion would be beneficial to both the practitioner and patient. The purpose of the study was to evaluate the weight of debris and volume of irrigant extruded apically from extracted teeth in vitro after endodontic instrumentation using four different rotary root canal instrumentation systems. Four groups of each 20 extracted mandibular premolars were instrumented using one of the four systems: ProTaper Universal (Dentsply Maillefer, Ballaigues, Switzerland)), Hero-shaper (MicroMega, Besancon, France), RaCe (FKG Dentaire, La-Chaux-de-Fonds, Switzerland) and K3 (SybronEndo, West Collins, CA). Debris and irrigant extruded from the apical foramen during instrumentation were collected in preweighed test tubes. Volume of irrigant extruded was noted. The containers were stored in incubator at 70° for two days to evaporate the moisture. Weight of dry debris was noted. Data was analyzed using Kruskall-Wallis and Mann-Whitney U test at a significance of 0.001. The results indicated that all of the instrumentation systems tested caused measurable apical extrusion of debris and irrigants. Higher extrusion was observed with Protaper system which was statistically significant with Hero-Shaper, RaCe and K3 systems. There were no statistical differences between Hero-shaper, K3 and RaCe systems (p < 0.05). All instrumentation techniques apically extruded debris and irrigant. However, Hero-shaper, K3 and RaCe systems produced less extruded debris and irrigant than the Protaper system.

  10. Escherichia coli Uropathogenesis In Vitro

    DEFF Research Database (Denmark)

    Andersen, Thomas E; Khandige, Surabhi; Madelung, Michelle

    2012-01-01

    -stage infection events have not been replicated in vitro. We have established an in vitro model of human bladder cell infection by the use of a flow chamber (FC)-based culture system, which allows investigation of steps subsequent to initial invasion. Short-term bacterial colonization on the FC-BEC layer led...... to rods that could invade other BECs. Hence, under growth conditions established to resemble those present in vivo, the elements of the proposed uropathogenic cascade were inducible in a human BEC model system. Here, we describe the model and show how these characteristics are reproduced in vitro....

  11. Development of a high-throughput in vitro assay using a novel Caco-2/rat hepatocyte system for the prediction of oral plasma area under the concentration versus time curve (AUC) in rats.

    Science.gov (United States)

    Cheng, K-C; Li, Cheng; Hsieh, Yunsheng; Montgomery, Diana; Liu, Tongtong; White, Ronald E

    2006-01-01

    Previously, we have shown that a novel Caco-2/human hepatocyte system is a useful model for the prediction of oral bioavailability in humans. In this study, we attempted to use a similar system in a high-throughput screening mode for the selection of new compound entities (NCE) in drug discovery. A total of 72 compounds randomly selected from three different chemotypes were dosed orally in rats. In vivo plasma area under the concentration versus time curve (AUC) from 0-6 h of the parent compound was determined. The same compounds were also tested in the Caco-2/rat hepatocyte system. In vitro AUC from 0-3 h in the Caco-2 rat hepatocyte system was determined. The predictive usefulness of the Caco-2/rat hepatocyte system was evaluated by comparing the in vivo plasma AUC and the in vitro AUC. Linear regression analysis showed a reasonable correlation (R2 = 0.5) between the in vivo AUC and the in vitro AUC. Using 0.4 microM h in vivo AUC as a cut-off, compounds were categorized as either low or high AUC. The in vitro AUC successfully matched the corresponding in vivo category for sixty-three out of seventy-two compounds. The results presented in this study suggest that the Caco-2/rat hepatocyte system may be used as a high-throughput screen in drug discovery for pharmacokinetic behaviors of compounds in rats.

  12. A reliable in vitro fruiting system for armillaria mellea for evaluation of agrobacterium tumefaciens transformation vectors

    Science.gov (United States)

    Armillaria mellea is a serious pathogen of horticultural and agricultural systems in Europe and North America. The lack of a reliable in vitro fruiting system has hindered research, and necessitated dependence on intermittently available wild-collected basidiospores. Here we describe a reliable, rep...

  13. Different test systems in Aspergillus nidulans for the evaluation of mitotic gene conversion, crossing-over and non-disjunction

    International Nuclear Information System (INIS)

    De Bertoldi, M.; Griselli, M.; Consiglio Nazionale delle Ricerche, Pisa; Barale, R.

    1980-01-01

    The wide variety of the genetic alterations produced by environmental mutagens has increased the necessity of using experimental microorganisms to reveal the induction of such genetic events with short-term tests. Aspergillus nidulans, because of its well-developed genetic system and the availability of morphological markers seay to score, can be profitably used in mutagen testing. The constitution of particular diploid strains of A. nidulans able to detect the induction of mitotic gene conversion, mitotic crossing-over and mitotic non-disjunction with selective procedures are described and validated with standard mutagens: methyl methanesulphonate and UV radiation (lacking a specific genetic activity), benomyl and p-fluorophenylalanine (with a specific genetic activity). The possibility of using mammalian metabolic activation of promutagens in A. nidulans in vitro was tested with cyclophosphamide, with positive results in all the tested genetic systems. A method that increases the sensitivity of conidia to mutagenic treatments is described; its application appeared to be particularly useful in experiments on crossing-over and non-disjunction. (orig.)

  14. Evaluation of in vitro culture systems for the maintenance of microfilariae and infective larvae of Loa loa.

    Science.gov (United States)

    Zofou, Denis; Fombad, Fanny Fri; Gandjui, Narcisse V T; Njouendou, Abdel Jelil; Kengne-Ouafo, Arnaud Jonas; Chounna Ndongmo, Patrick W; Datchoua-Poutcheu, Fabrice R; Enyong, Peter A; Bita, Dizzle Tayong; Taylor, Mark J; Turner, Joseph D; Wanji, Samuel

    2018-05-02

    Suitable and scalable in vitro culture conditions for parasite maintenance are needed to foster drug research for loiasis, one of the neglected tropical diseases which has attracted only limited attention over recent years, despite having important public health impacts. The present work aims to develop adequate in vitro culture systems for drug screening against both microfilariae (mf) and infective third-stage larvae (L3) of Loa loa. In vitro culture conditions were evaluated by varying three basic culture media: Roswell Park Memorial Institute (RPMI-1640), Dulbecco's modified Eagle's medium (DMEM) and Iscove's modified Dulbecco's medium (IMDM); four sera/proteins: newborn calf serum (NCS), foetal bovine serum (FBS), bovine serum albumin (BSA) and the lipid-enriched BSA (AlbuMax® II, ALB); and co-culture with the Monkey Kidney Epithelial Cell line (LLC-MK2) as a feeder layer. The various culture systems were tested on both mf and L3, using survival (% motile), motility (T 90 = mean duration (days) at which at least 90% of parasites were fully active) and moulting rates of L3 as the major criteria. The general linear model regression analysis was performed to assess the contribution of each variable on the viability of Loa loa L3 and microfilarie. All statistical tests were performed at 95% confidence interval. Of the three different media tested, DMEM and IMDM were the most suitable sustaining the maintenance of both L. loa L3 and mf. IMDM alone could sustain L3 for more than 5 days (T 90 = 6.5 ± 1.1 day). Serum supplements and LLC-MK2 co-cultures significantly improved the survival of parasites in DMEM and IMDM. In co-cultures with LLC-MK2 cells, L. loa mf were maintained in each of the three basic media (T 90 of 16.4-19.5 days) without any serum supplement. The most effective culture systems promoting significant moulting rate of L3 into L4 (at least 25%) with substantial maintenance time were: DMEM + BSA, DMEM + NCS, DMEM-AlbuMax®II, DMEM + FBS all in co

  15. Forecasting gastrointestinal precipitation and oral pharmacokinetics of dantrolene in dogs using an in vitro precipitation testing coupled with in silico modeling and simulation.

    Science.gov (United States)

    Kambayashi, Atsushi; Dressman, Jennifer B

    2017-10-01

    The aim of the current research was to determine the precipitation kinetics of dantrolene sodium using canine biorelevant in vitro testing and to model the precipitation kinetics by appropriately coupling the data with an in silico tool adapted for dogs. The precipitation profiles of dantrolene sodium solutions were obtained with the in vitro paddle apparatus at a revolution rate of 50rpm. The in silico prediction tool was designed using STELLA software and the predicted plasma concentration profiles of dantrolene using the in vitro precipitation data were compared with the observed in vivo pharmacokinetics in beagle dogs. The plasma profiles of dantrolene, which served as a model weakly acidic drug which precipitates in the upper gastrointestinal tract, was successfully predicted using the in vitro precipitation testing coupled with the in silico modeling and simulation approach. The approach was subsequently used to forecast the effect of pharmaceutical excipients (HPMC/PG) on the ability of the drug to supersaturate in the gut and the resulting pharmacokinetics. The agreement of the simulated pharmacokinetics with the observed values confirms the ability of canine biorelevant media to predict oral performance of enhanced dosage forms in dogs. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. [Elaboration of Pseudo-natural Products Using Artificial In Vitro Biosynthesis Systems].

    Science.gov (United States)

    Goto, Yuki

    2018-01-01

     Peptidic natural products often consist of not only proteinogenic building blocks but also unique non-proteinogenic structures such as macrocyclic scaffolds and N-methylated backbones. Since such non-proteinogenic structures are important structural motifs that contribute to diverse bioactivity, we have proposed that peptides with non-proteinogenic structures should be attractive candidates as artificial bioactive peptides mimicking natural products, or so-called pseudo-natural products. We previously devised an engineered translation system for pseudo-natural peptides, referred to as the flexible in vitro translation (FIT) system. This system enabled "one-pot" synthesis of highly diverse pseudo-natural peptide libraries, which can be rapidly screened by mRNA display technology for the discovery of pseudo-natural peptides with diverse bioactivities.

  17. An In Vitro System Comprising Immortalized Hypothalamic Neuronal Cells (GT1–7 Cells for Evaluation of the Neuroendocrine Effects of Essential Oils

    Directory of Open Access Journals (Sweden)

    Dai Mizuno

    2015-01-01

    Full Text Available Aromatherapy and plant-based essential oils are widely used as complementary and alternative therapies for symptoms including anxiety. Furthermore, it was reportedly effective for the care of several diseases such as Alzheimer’s disease and depressive illness. To investigate the pharmacological effects of essential oils, we developed an in vitro assay system using immortalized hypothalamic neuronal cells (GT1–7 cells. In this study, we evaluated the effects of essential oils on neuronal death induced by hydrogen peroxide (H2O2, aluminum, zinc, or the antagonist of estrogen receptor (tamoxifen. Among tests of various essential oils, we found that H2O2-induced neuronal death was attenuated by the essential oils of damask rose, eucalyptus, fennel, geranium, ginger, kabosu, mandarin, myrrh, and neroli. Damask rose oil had protective effects against aluminum-induced neurotoxicity, while geranium and rosemary oil showed protective activity against zinc-induced neurotoxicity. In contrast, geranium oil and ginger oil enhanced the neurotoxicity of tamoxifen. Our in vitro assay system could be useful for the neuropharmacological and endocrine pharmacological studies of essential oils.

  18. In vitro disposition profiling of heterocyclic compounds.

    Science.gov (United States)

    Keemink, Janneke; Wuyts, Benjamin; Nicolaï, Johan; Jonghe, Steven De; Stella, Alessandro; Herdewijn, Piet; Augustijns, Patrick; Annaert, Pieter

    2015-08-01

    Compound libraries that are screened for biological activity commonly contain heterocycles. Besides potency, drug-like properties need to be evaluated to ensure in vivo efficacy of test compounds. In this context, we determined hepatic and intestinal disposition profiles for 17 heterocyclic compounds. All studied compounds showed rapid uptake in suspended rat hepatocytes, whereas metabolism was poor and the rate-limiting step in hepatic elimination. In vitro assays demonstrated a relatively low solubility and high intestinal permeability. Based on these in vitro data, heterocycles were categorized in the biopharmaceutics classification system (BCS) and the biopharmaceutics drug disposition classification system (BDDCS) to predict disposition characteristics before clinical data are available. Our findings emphasized the importance to use hepatocytes in addition to microsomes to study metabolism, since the latter lack non-microsomal enzymes and cellular context. Moreover, intracellular exposure should be considered to gain insight in the relevant fraction of the compound available at the enzymatic site. Finally, the study reveals discrepancies associated with the classification of heterocycles in BCS versus BDDCS. These probably originate from the binary character of both systems. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. In vitro evaluation of transdermal nicotine delivery systems commercially available in Brazil

    Directory of Open Access Journals (Sweden)

    André Luís Morais Ruela

    2013-09-01

    Full Text Available The aim of this study was to develop and validate a method for evaluating the release and skin permeation from transdermal nicotine patches using the vertical diffusion cell (VDC. The VDC is an experimental apparatus employed in research, development, and the pharmaceutical field because it can simulate conditions closest to those established in clinical trials. Two transdermal nicotine delivery systems marketed in Brazil to release 14 mg over 24 hours were evaluated. Release studies were carried out using a regenerated cellulose dialysis membrane and permeation studies were carried out using excised porcine ear skin. The results indicated that nicotine release from both evaluated patches follows Higuchi's release kinetics, while skin permeation studies indicated zero-order release kinetics. Nicotine release rates were different between both evaluated patches, but drug permeation rates were not significantly different. According to validation studies, the method was appropriate for evaluating in vitro performance of nicotine patches. The proposed method can be applied to in vitro comparative studies between different commercial nicotine patches and may be used as an auxiliary tool in the design of new transdermal nicotine delivery systems.

  20. Selection of potential probiotic lactic acid bacteria from fermented olives by in vitro tests.

    Science.gov (United States)

    Argyri, Anthoula A; Zoumpopoulou, Georgia; Karatzas, Kimon-Andreas G; Tsakalidou, Effie; Nychas, George-John E; Panagou, Efstathios Z; Tassou, Chrysoula C

    2013-04-01

    The present study aims to evaluate the probiotic potential of lactic acid bacteria (LAB) isolated from naturally fermented olives and select candidates to be used as probiotic starters for the improvement of the traditional fermentation process and the production of newly added value functional foods. Seventy one (71) lactic acid bacterial strains (17 Leuconostoc mesenteroides, 1 Ln. pseudomesenteroides, 13 Lactobacillus plantarum, 37 Lb. pentosus, 1 Lb. paraplantarum, and 2 Lb. paracasei subsp. paracasei) isolated from table olives were screened for their probiotic potential. Lb. rhamnosus GG and Lb. casei Shirota were used as reference strains. The in vitro tests included survival in simulated gastrointestinal tract conditions, antimicrobial activity (against Listeria monocytogenes, Salmonella Enteritidis, Escherichia coli O157:H7), Caco-2 surface adhesion, resistance to 9 antibiotics and haemolytic activity. Three (3) Lb. pentosus, 4 Lb. plantarum and 2 Lb. paracasei subsp. paracasei strains demonstrated the highest final population (>8 log cfu/ml) after 3 h of exposure at low pH. The majority of the tested strains were resistant to bile salts even after 4 h of exposure, while 5 Lb. plantarum and 7 Lb. pentosus strains exhibited partial bile salt hydrolase activity. None of the strains inhibited the growth of the pathogens tested. Variable efficiency to adhere to Caco-2 cells was observed. This was the same regarding strains' susceptibility towards different antibiotics. None of the strains exhibited β-haemolytic activity. As a whole, 4 strains of Lb. pentosus, 3 strains of Lb. plantarum and 2 strains of Lb. paracasei subsp. paracasei were found to possess desirable in vitro probiotic properties similar to or even better than the reference probiotic strains Lb. casei Shirota and Lb. rhamnosus GG. These strains are good candidates for further investigation both with in vivo studies to elucidate their potential health benefits and in olive fermentation processes

  1. Minicomputer controlled test system for process control and monitoring systems

    International Nuclear Information System (INIS)

    Worster, L.D.

    A minicomputer controlled test system for testing process control and monitoring systems is described. This system, in service for over one year, has demonstrated that computerized control of such testing has a real potential for expanding the scope of the testing, improving accuracy of testing, and significantly reducing the time required to do the testing. The test system is built around a 16-bit minicomputer with 12K of memory. The system programming language is BASIC with the addition of assembly level routines for communication with the peripheral devices. The peripheral devices include a 100 channel scanner, analog-to-digital converter, visual display, and strip printer. (auth)

  2. Inter-laboratory study of human in vitro toxicogenomics-based tests as alternative methods for evaluating chemical carcinogenicity : a bioinformatics perspective

    NARCIS (Netherlands)

    Herwig, R; Gmuender, H; Corvi, Raffaella; Bloch, K.M.; Brandenburg, A.; Castell, J; Ceelen, L; Chesne, C; Doktorova, T Y; Jennen, D; Jennings, P; Limonciel, A; Lock, E A; McMorrow, T; Phrakonkham, P; Radford-Smith, G.; Slattery, C; Stierum, R; Vilardell, M; Wittenberger, T; Yildirimman, R; Ryan, M.; Rogiers, Vera; Kleinjans, Jos

    The assessment of the carcinogenic potential of chemicals with alternative, human-based in vitro systems has become a major goal of toxicogenomics. The central read-out of these assays is the transcriptome, and while many studies exist that explored the gene expression responses of such systems,

  3. In vitro and in vivo performance of monoacyl phospholipid-based self-emulsifying drug delivery systems

    DEFF Research Database (Denmark)

    Tran, Thuy; Siqueira, Scheyla D V S; Amenitsch, Heinz

    2017-01-01

    to investigate the impact of LPC on the emulsion droplet size, extent of digestion, colloidal structure evolution and drug precipitation during in vitro lipolysis simulating human conditions and drug bioavailability in a rat model. The four investigated SEDDS containing long-chain glycerides, polyoxyl 35 castor...... and polydispersity of the dispersed SEDDS. The two LPC-free SEDDS generated lamellar phase structures (Lα) with d-spacing=4.76nm during digestion. Incorporating LPC into these systems inhibited the formation of lamellar phase structures. The amount of precipitated crystalline FF from the four SEDDS was similar......This study investigates the effect of monoacyl phospholipid incorporation on the in vitro and in vivo performance of self-emulsifying drug delivery systems (SEDDS). Monoacyl phosphatidylcholine (Lipoid S LPC 80 (LPC)) was incorporated into four different fenofibrate (FF)-loaded long-chain SEDDS...

  4. The antioxidant system of seminal fluid during in vitro storage of sterlet Acipenser ruthenus sperm.

    Science.gov (United States)

    Dzyuba, Viktoriya; Cosson, Jacky; Dzyuba, Borys; Yamaner, Gunes; Rodina, Marek; Linhart, Otomar

    2016-04-01

    The role of the seminal fluid antioxidant system in protection against damage to spermatozoa during in vitro sperm storage is unclear. This study investigated the effect of in vitro storage of sterlet Acipenser ruthenus spermatozoa together with seminal fluid for 36 h at 4 °C on spermatozoon motility rate and curvilinear velocity, thiobarbituric acid reactive substance level, and components of enzyme and non-enzyme antioxidant system (superoxide dismutase and catalase activity and uric acid concentration) in seminal fluid. Spermatozoon motility parameters after sperm storage were significantly decreased, while the level of thiobarbituric acid reactive substances, activity of superoxide dismutase and catalase, and uric acid concentration did not change. Our findings suggest that the antioxidant system of sterlet seminal fluid is effective in preventing oxidative stress during short-term sperm storage and prompt future investigations of changes in spermatozoon homeostasis and in spermatozoon plasma membrane structure which are other possible reasons of spermatozoon motility deterioration upon sperm storage.

  5. In vitro evaluation of marginal discrepancy of monolithic zirconia restorations fabricated with different CAD-CAM systems.

    Science.gov (United States)

    Hamza, Tamer A; Sherif, Rana M

    2017-06-01

    Dental laboratories use different computer-aided design and computer-aided manufacturing (CAD-CAM) systems to fabricate fixed prostheses; however, limited evidence is available concerning which system provides the best marginal discrepancy. The purpose of this in vitro study was to evaluate the marginal fit of 5 different monolithic zirconia restorations milled with different CAD-CAM systems. Thirty monolithic zirconia crowns were fabricated on a custom-designed stainless steel die and were divided into 5 groups according to the type of monolithic zirconia crown and the CAD-CAM system used: group TZ, milled with an MCXL milling machine; group CZ, translucent zirconia milled with a motion milling machine; group ZZ, zirconia milled with a dental milling unit; group PZ, translucent zirconia milled with a zirconia milling unit; and group BZ, solid zirconia milled using an S1 VHF milling machine. The marginal fit was measured with a binocular microscope at an original magnification of ×100. The results were tabulated and statistically analyzed with 1-way ANOVA and post hoc surface range test, and pairwise multiple comparisons were made using Bonferroni correction (α=.05). The type of CAD-CAM used affected the marginal fit of the monolithic restoration. The mean (±SD) highest marginal discrepancy was recorded in group TZI at 39.3 ±2.3 μm, while the least mean marginal discrepancy was recorded in group IZ (22.8 ±8.9 μm). The Bonferroni post hoc test showed that group TZI was significantly different from all other groups tested (Pmarginal discrepancies; however, the CAD-CAM system with the 5-axis milling unit produced the best marginal fit. Copyright © 2016 Editorial Council for the Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.

  6. Development of an integrated electrochemical system for in vitro yeast viability testing.

    Science.gov (United States)

    Adami, Andrea; Ress, Cristina; Collini, Cristian; Pedrotti, Severino; Lorenzelli, Leandro

    2013-02-15

    This work describes the development and testing of a microfabricated sensor for rapid cell growth monitoring, especially focused on yeast quality assessment for wine applications. The device consists of a NMOS ISFET sensor with Si(3)N(4) gate, able to indirectly monitor extracellular metabolism through pH variation of the medium, and a solid-state reference electrode implemented with PVC membranes doped with lipophilic salts (tetrabutylammonium-tetrabutylborate (TBA-TBB) and Potassium tetrakis(4-chlorphenyl)borate (KTClpB)). The use of a solid state reference electrode enables the implementation of a large number of cell assays in parallel, without the need of external conventional reference electrodes. Microbial growth testing has been performed both in standard culture conditions and on chip at different concentrations of ethanol in order to carry out a commonly used screening of wine yeast strains. Cell growth tests can be performed in few hours, providing a fast, sensitive and low cost analysis with respect to the conventional procedures. Copyright © 2012 Elsevier B.V. All rights reserved.

  7. Validation of capillary blood analysis and capillary testing mode on the epoc Point of Care system

    Directory of Open Access Journals (Sweden)

    Jing Cao

    2017-12-01

    Full Text Available Background: Laboratory test in transport is a critical component of patient care, and capillary blood is a preferred sample type particularly in children. This study evaluated the performance of capillary blood testing on the epoc Point of Care Blood Analysis System (Alere Inc. Methods: Ten fresh venous blood samples was tested on the epoc system under the capillary mode. Correlation with GEM 4000 (Instrumentation Laboratory was examined for Na+, K+, Cl-, Ca2+, glucose, lactate, hematocrit, hemoglobin, pO2, pCO2, and pH, and correlation with serum tested on Vitros 5600 (Ortho Clinical Diagnostics was examined for creatinine. Eight paired capillary and venous blood was tested on epoc and ABL800 (Radiometer for the correlation of Na+, K+, Cl-, Ca2+, glucose, lactate, hematocrit, hemoglobin, pCO2, and pH. Capillary blood from 23 apparently healthy volunteers was tested on the epoc system to assess the concordance to reference ranges used locally. Results: Deming regression correlation coefficients for all the comparisons were above 0.65 except for ionized Ca2+. Accordance of greater than 85% to the local reference ranges were found in all assays with the exception of pO2 and Cl-. Conclusion: Data from this study indicates that capillary blood tests on the epoc system provide comparable results to reference method for these assays, Na+, K+, glucose, lactate, hematocrit, hemoglobin, pCO2, and pH. Further validation in critically ill patients is needed to implement the epoc system in patient transport. Impact of the study: This study demonstrated that capillary blood tests on the epoc Point of Care Blood Analysis System give comparable results to other chemistry analyzers for major blood gas and critical tests. The results are informative to institutions where pre-hospital and inter-hospital laboratory testing on capillary blood is a critical component of patient point of care testing. Keywords: Epoc, Capillary, Transport, Blood gas, Point of care

  8. Preclinical Testing of the Nitroimidazopyran PA-824 for Activity against Mycobacterium tuberculosis in a Series of In Vitro and In Vivo Models

    OpenAIRE

    Lenaerts, Anne J.; Gruppo, Veronica; Marietta, Karen S.; Johnson, Christine M.; Driscoll, Diane K.; Tompkins, Nicholas M.; Rose, Jerry D.; Reynolds, Robert C.; Orme, Ian M.

    2005-01-01

    This study extends earlier reports regarding the in vitro and in vivo efficacies of the nitroimidazopyran PA-824 against Mycobacterium tuberculosis. PA-824 was tested in vitro against a broad panel of multidrug-resistant clinical isolates and was found to be highly active against all isolates (MIC < 1 μg/ml). The activity of PA-824 against M. tuberculosis was also assessed grown under conditions of oxygen depletion. PA-824 showed significant activity at 2, 10, and 50 μg/ml, similar to that of...

  9. A reassessment of the in vitro RBC haemolysis assay with defibrinated sheep blood for the determination of the ocular irritation potential of cosmetic products: comparison with the in vivo Draize rabbit test.

    Science.gov (United States)

    Alves, Eloísa Nunes; Presgrave, Rosaura de Farias; Presgrave, Octávio Augusto França; Sabagh, Fernanda Peres; de Freitas, João Carlos Borges Rolim; Corrado, Alexandre P

    2008-07-01

    We examined the correlation between results obtained from the in vivo Draize test for ocular irritation and in vitro results obtained from the sheep red blood cell (RBC) haemolytic assay, which assesses haemolysis and protein denaturation in erythrocytes, induced by cosmetic products. We sought to validate the haemolytic assay as a preliminary test for identifying highly-irritative products, and also to evaluate the in vitro test as alternative assay for replacement of the in vivo test. In vitro and in vivo analyses were carried out on 19 cosmetic products, in order to correlate the lesions in the ocular structures with three in vitro parameters: (i) the extent of haemolysis (H50); (ii) the protein denaturation index (DI); and (iii) the H50/DI ratio, which reflects the irritation potential (IP). There was significant correlation between maximum average scores (MAS) and the parameters determined in vitro (r = 0.752-0.764). These results indicate that the RBC assay is a useful and rapid test for use as a screening method to assess the IP of cosmetic products, and for predicting the IP value with a high level of concordance (94.7%). The assay showed high sensitivity and specificity rates of 91.6% and 100%, respectively.

  10. RIVETS: a mechanical system for in vivo and in vitro electrophysiology and imaging.

    Directory of Open Access Journals (Sweden)

    Jason E Osborne

    Full Text Available A number of recent studies have provided compelling demonstrations that both mice and rats can be trained to perform a variety of behavioral tasks while restrained by mechanical elements mounted to the skull. The independent development of this technique by a number of laboratories has led to diverse solutions. We found that these solutions often used expensive materials and impeded future development and modification in the absence of engineering support. In order to address these issues, here we report on the development of a flexible single hardware design for electrophysiology and imaging both in brain tissue in vitro. Our hardware facilitates the rapid conversion of a single preparation between physiology and imaging system and the conversion of a given system between preparations. In addition, our use of rapid prototyping machines ("3D printers" allows for the deployment of new designs within a day. Here, we present specifications for design and manufacturing as well as some data from our lab demonstrating the suitability of the design for physiology in behaving animals and imaging in vitro and in vivo.

  11. Fatigue tests of dowel-socket systems

    International Nuclear Information System (INIS)

    Chiang, D.D.

    1976-01-01

    A test program was conducted to determine the fatigue behavior of LHTGR fuel element dowel/socket systems. Two dowel/socket systems, namely, a four-dowel system and a five-dowel system, were tested to failure under shear loads applied through a fatigue test apparatus to simulate repetitive loading during a seismic event

  12. The Effect of Canal Preparation with Four Different Rotary Systems on Formation of Dentinal Cracks: An In Vitro Evaluation.

    Science.gov (United States)

    Khoshbin, Elham; Donyavi, Zakiyeh; Abbasi Atibeh, Erfan; Roshanaei, Ghodratollah; Amani, Faranak

    2018-01-01

    Endodontic rotary systems may result in dentinal cracks. They may propagate to vertical root fracture that compromises the outcome of endodontic treatment. This study aimed to compare Neolix and Reciproc (single-file systems), Mtwo and ProTaper (conventional rotary systems) in terms of dentinal crack formation in root canal walls. This in vitro study was conducted on 110 extracted human single-rooted teeth. The teeth were randomly divided into four experimental groups ( n =25) for root canal preparation with Neolix, Reciproc, Mtwo and ProTaper systems and two control groups ( n =5). The first control group underwent root canal instrumentation with hand files while the second control group received no preparation and was only irrigated. After instrumentation, root canals were horizontally sectioned at 3, 6 and 9 mm from the apex and inspected under a stereomicroscope under 12× magnification for detection of cracks. The data were analyzed using Chi-square, GEE test and Bonferroni tests ( P ProTaper, Reciproc, Mtwo and Neolix caused cracks in 92%, 80%, 68% and 48% of samples. ProTaper caused significantly more cracks than Neolix and Mtwo ( P 0.05). All rotary systems cause dentinal cracks and it is significantly different in apical, middle and coronal third of the root. Neolix appears to be a suitable alternative to other rotary systems since use of this single-file system saves time and cost and minimizes trauma to dentinal walls.

  13. System Performance and Testing

    NARCIS (Netherlands)

    Frei, U.; Oversloot, H.

    2004-01-01

    This chapter compares and contrasts the system performance of two widely used solar thermal systems using testing and simulation programs. Solar thermal systems are used in many countries for heating domestically used water. In addition to the simple thermosiphon systems, better designed pumped

  14. Effects of Achyrocline satureioides Inflorescence Extracts against Pathogenic Intestinal Bacteria: Chemical Characterization, In Vitro Tests, and In Vivo Evaluation

    Directory of Open Access Journals (Sweden)

    Karla Suzana Moresco

    2017-01-01

    Full Text Available Three Achyrocline satureioides (AS inflorescences extracts were characterized: (i a freeze-dried extract prepared from the aqueous extractive solution and (ii a freeze-dried and (iii a spray-dried extract prepared from hydroethanol extractive solution (80% ethanol. The chemical profile, antioxidant potential, and antimicrobial activity against intestinal pathogenic bacteria of AS extracts were evaluated. In vitro antioxidant activity was determined by the total reactive antioxidant potential (TRAP assay. In vivo analysis and characterization of intestinal microbiota were performed in male Wistar rats (saline versus treated animals with AS dried extracts by high-throughput sequencing analysis: metabarcoding. Antimicrobial activity was tested in vitro by the disc diffusion tests. Moisture content of the extracts ranged from 10 to 15% and 5.7 to 17 mg kg−1 of fluorine. AS exhibited antioxidant activity, especially in its freeze-dried form which also exhibited a wide spectrum of antimicrobial activity against intestinal pathogenic bacteria greater than those observed by the antibiotic, amoxicillin, when tested against Bacillus cereus and Staphylococcus aureus. Antioxidant and antimicrobial activities of AS extracts seemed to be positively correlated with the present amount of flavonoids. These findings suggest a potential use of AS as a coadjuvant agent for treating bacterial-induced intestinal diseases with high rates of antibiotic resistance.

  15. In vitro and in vivo percutaneous absorption of retinol from cosmetic formulations: Significance of the skin reservoir and prediction of systemic absorption

    International Nuclear Information System (INIS)

    Yourick, Jeffrey J.; Jung, Connie T.; Bronaugh, Robert L.

    2008-01-01

    The percutaneous absorption of retinol (Vitamin A) from cosmetic formulations was studied to predict systemic absorption and to understand the significance of the skin reservoir in in vitro absorption studies. Viable skin from fuzzy rat or human subjects was assembled in flow-through diffusion cells for in vitro absorption studies. In vivo absorption studies using fuzzy rats were performed in glass metabolism cages for collection of urine, feces, and body content. Retinol (0.3%) formulations (hydroalcoholic gel and oil-in-water emulsion) containing 3 H-retinol were applied and absorption was measured at 24 or 72 h. All percentages reported are % of applied dose. In vitro studies using human skin and the gel and emulsion vehicles found 0.3 and 1.3% retinol, respectively, in receptor fluid at 24 h. Levels of absorption in the receptor fluid increased over 72 h with the gel and emulsion vehicles. Using the gel vehicle, in vitro rat skin studies found 23% in skin and 6% in receptor fluid at 24 h, while 72-h studies found 18% in skin and 13% in receptor fluid. Thus, significant amounts of retinol remained in rat skin at 24 h and decreased over 72 h, with proportional increases in receptor fluid. In vivo rat studies with the gel found 4% systemic absorption of retinol after 24 h and systemic absorption did not increase at 72 h. Retinol remaining in rat skin after in vivo application was 18% and 13% of the applied dermal dose after 24 and 72 h, respectively. Similar observations were made with the oil-in water emulsion vehicle in the rat. Retinol formed a reservoir in rat skin both in vivo and in vitro. Little additional retinol was bioavailable after 24 h. Comparison of these in vitro and in vivo results for absorption through rat skin indicates that the 24-h in vitro receptor fluid value accurately estimated 24-h in vivo systemic absorption. Therefore, the best single estimate of retinol systemic absorption from in vitro human skin studies is the 24-h receptor fluid

  16. The 13th quality control survey for radioisotopes in vitro tests in Japan, 1991

    Energy Technology Data Exchange (ETDEWEB)

    1992-11-01

    This paper presents the results of the 13th quality control nationwide survey. Of 568 hospitals performing RI in vitro tests as of December 1991, 280 (49.3%) participated in this survey. The following 37 items were examined: ACTH, 17[alpha]-hydroxyprogesterone, [alpha]-fetoprotein, aldosterone, [beta]2-microglobulin, CA 125, CA 15-3, CA 19-9, calcitonin, CEA, cortisol, C-peptide, digoxin, esteradiol, ferritin, free triiodothyronine (T[sub 3]), free thyroxine (T[sub 4]), FSH, gastrin, GH, glucagon, IgE, insulin, LH, PAP, progesterone, prolactin, PTH, somatomedin C, T[sub 3], T[sub 4], T[sub 3] uptake, TBG, testosterone, thyroglobulin, tissue polypeptide antigen, and TSH. Reliability and safety of measurement values were evaluated based on protein components of serum samples, spontaneous reliability test, time-dependent reliability test, repeated freezing-dissolution test, and serial change test after freezing. 'Within kit variation' between facilities was great especially for ACTH, CA 15-3, CA 19-9, free T[sub 4], calcitonin, gastrin, and testosterone, and was favorable for [beta][sub 2]-microglobulin, CA 125, digoxin, cortisol, IgE, T[sub 3], T[sub 4], T[sub 3] uptake, and TBG. It was noted that there was a great difference in measurement values of free T[sub 3] and free-T[sub 4] between RIA kits and non-isotopic kits. (N.K.).

  17. In vitro growth and leaf anatomy of Cattleya walkeriana (Gardner, 1839 grown in natural ventilation system

    Directory of Open Access Journals (Sweden)

    Adriano Bortolotti da Silva

    2014-12-01

    Full Text Available Natural ventilation system facilitates gaseous exchanges in in vitro plants promoting changes in the leaf tissue, which can be evaluated through the leaf anatomy, and it allows a cultivation closer to the photoautrophic micropropagation. The objective of this work was to evaluate the effects on in vitro growth and on the leaf anatomy of Cattleya walkeriana grown in natural and conventional ventilation system with different concentrations of sucrose (0; 15; 30 and 45 L-1 combined with different cultivation systems (conventional micropropagation and natural ventilation system. The culture medium was composed of MS salts, solidified with 7 g L-1 of agar and pH adjusted to 5.8. Forty milliliters of culture medium were distributed in 250 mL flasks, autoclaved at 120 ºC for 20 minutes. The greater plant growth, as well as the greater thickness of the mesophyll was observed with the use of 20 g L-1 sucrose in natural ventilation system. Plants grown in natural ventilation system showed a thicker leaf mesophyll, which is directly related to photoautotrophic crops. The natural ventilation system induced more elliptical stomata and probably more functional formats.

  18. Liquid crystalline systems for transdermal delivery of celecoxib: in vitro drug release and skin permeation studies.

    Science.gov (United States)

    Estracanholli, Eder André; Praça, Fabíola Silva Garcia; Cintra, Ana Beatriz; Pierre, Maria Bernadete Riemma; Lara, Marilisa Guimarães

    2014-12-01

    Liquid crystalline systems of monoolein/water could be a promising approach for the delivery of celecoxib (CXB) to the skin because these systems can sustain drug release, improve drug penetration into the skin layers and minimize side effects. This study evaluated the potential of these systems for the delivery of CXB into the skin based on in vitro drug release and skin permeation studies. The amount of CXB that permeated into and/or was retained in the skin was assayed using an HPLC method. Polarizing light microscopy studies showed that liquid crystalline systems of monoolein/water were formed in the presence of CXB, without any changes in the mesophases. The liquid crystalline systems decreased drug release when compared to control solution. Drug release was independent of the initial water content of the systems and CXB was released from cubic phase systems, irrespective of the initial water content. The systems released the CXB following zero-order release kinetics. In vitro drug permeation studies showed that cubic phase systems allowed drug permeation and retention in the skin layers. Cubic phase systems of monoolein/water may be promising vehicles for the delivery of CXB in/through the skin because it improved CXB skin permeation compared with the control solution.

  19. Establishment of a new immortalized human corneal epithelial cell line (iHCE-NY1) for use in evaluating eye irritancy by in vitro test methods.

    Science.gov (United States)

    Yamamoto, Naoki; Kato, Yoshinao; Sato, Atsushi; Hiramatsu, Noriko; Yamashita, Hiromi; Ohkuma, Mahito; Miyachi, Ei-Ichi; Horiguchi, Masayuki; Hirano, Koji; Kojima, Hajime

    2016-08-01

    In vitro test methods that use human corneal epithelial cells to evaluate the eye irritation potency of chemical substances do not use human corneal epithelium because it has been difficult to maintain more than four passages. In this study, we make a new cell line comprising immortalized human corneal epithelial cells (iHCE-NY1). The IC50 of iHCE-NY1 cells is slightly higher than that of Statens Seruminstitut Rabbit Cornea (SIRC) cells, which are currently used in some in vitro test methods. CDKN1A in iHCE-NY1 cells was used as a marker of gene expression to indicate cell cycle activity. This enabled us to evaluate cell recovery characteristics at concentrations lower than the IC50 of cytotoxic tests.

  20. Effect of the well of the well (WOW) system on in vitro culture for porcine embryos after intracytoplasmic sperm injection.

    Science.gov (United States)

    Taka, Mikiko; Iwayama, Hiroshi; Fukui, Yutaka

    2005-08-01

    For developmental competence of porcine embryos in vitro, it is important to improve the culture environment. The present study was performed to evaluate four different culture systems for in vitro matured porcine oocytes following intracytoplasmic sperm injection (ICSI); drop, well and two sizes of the well of the well (WOW) systems (500 and 1,000 microm in diameter). The cleavage rate on Day 2 and the mean cell number in blastocysts on Day 6 were not significantly different among the four treatments. However, the 1,000 microm WOW (24.6%) resulted in a significantly higher (PWOW, respectively). The present study indicates that the microenvironment created by the 1,000 microm diameter WOW improves blastocyst production of in vitro matured porcine oocytes after ICSI, and that the effectiveness of the WOW system is dependent on the size (diameter) of the WOW.

  1. Arabidopsis Pol II-Dependent in Vitro Transcription System Reveals Role of Chromatin for Light-Inducible rbcS Gene Transcription1

    Science.gov (United States)

    Ido, Ayaka; Iwata, Shinya; Iwata, Yuka; Igarashi, Hisako; Hamada, Takahiro; Sonobe, Seiji; Sugiura, Masahiro; Yukawa, Yasushi

    2016-01-01

    In vitro transcription is an essential tool to study the molecular mechanisms of transcription. For over a decade, we have developed an in vitro transcription system from tobacco (Nicotiana tabacum)-cultured cells (BY-2), and this system supported the basic activities of the three RNA polymerases (Pol I, Pol II, and Pol III). However, it was not suitable to study photosynthetic genes, because BY-2 cells have lost their photosynthetic activity. Therefore, Arabidopsis (Arabidopsis thaliana) in vitro transcription systems were developed from green and etiolated suspension cells. Sufficient in vitro Pol II activity was detected after the minor modification of the nuclear soluble extracts preparation method; removal of vacuoles from protoplasts and L-ascorbic acid supplementation in the extraction buffer were particularly effective. Surprisingly, all four Arabidopsis Rubisco small subunit (rbcS-1A, rbcS-1B, rbcS-2B, and rbcS-3B) gene members were in vitro transcribed from the naked DNA templates without any light-dependent manner. However, clear light-inducible transcriptions were observed using chromatin template of rbcS-1A gene, which was prepared with a human nucleosome assembly protein 1 (hNAP1) and HeLa histones. This suggested that a key determinant of light-dependency through the rbcS gene transcription was a higher order of DNA structure (i.e. chromatin). PMID:26662274

  2. Analysis of Chemical Bioactivity through In Vitro Profiling ...

    Science.gov (United States)

    Safety assessment of drugs and environmental chemicals relies extensively on animal testing. However, the quantity of chemicals needing assessment and challenges of species extrapolation drive the development of alternative approaches. The EPA’s ToxCast and the multiagency Tox21 programs address this through use of an extensive in vitro screening program to generate data on a large library of important environmental chemicals. These in vitro assays encompass both cell-free, biochemical assays targeting proteins that may be potential molecular initiating events and cellular assays that provide coverage of critical signaling pathways and toxicity phenotypes. Effects on model organisms such as the developing zebrafish, are also part of the testing strategy. A variety of computational approaches are used to analyze the resulting complex data sets to gain insight in to inherent biological activity of chemicals and possible mechanisms of toxicity. Several case studies including identification of modulators of estrogen receptor and aromatic hydrocarbon receptor pathways with effects in primary human cell systems will be described. In addition, existing in vivo data from a subset of the chemicals was used to anchor predictive models using in vitro data for a number of adverse endpoints including reproductive and developmental toxicities. The strengths and weaknesses of this approach will be described. This work does not necessarily reflect official Agency policy. Pres

  3. Avaliação do sistema de monitoramento computadorizado de digestão in vitro: 1. Testes preliminares Evaluation of a computerized monitoring on in vitro digestion system: 1. Preliminary tests

    OpenAIRE

    Fábio Prudêncio de Campos; Dante Pazzanese Duarte Lanna; Máx Lázaro Vieira Bose; Celso Boin

    2000-01-01

    O objetivo deste trabalho foi testar o sistema de monitoramento computadorizado da produção de gás in vitro. Essa técnica facilita estudos da degradação das frações solúveis e insolúveis das forragens, quantificadas pela produção de gás (CO2 e CH4) oriunda do metabolismo microbiano e medida por sensor de pressão. Diversas quantidades de amostras (50 a 110 mg de feno de alfafa) e outros alimentos foram testadas. Também, a influência da quantidade de líquido ruminal (2,0 ou 3,0 mL), com ou sem ...

  4. Differentiating human NT2/D1 neurospheres as a versatile in vitro 3D model system for developmental neurotoxicity testing

    International Nuclear Information System (INIS)

    Hill, E.J.; Woehrling, E.K.; Prince, M.; Coleman, M.D.

    2008-01-01

    Developmental neurotoxicity is a major issue in human health and may have lasting neurological implications. In this preliminary study we exposed differentiating Ntera2/clone D1 (NT2/D1) cell neurospheres to known human teratogens classed as non-embryotoxic (acrylamide), weakly embryotoxic (lithium, valproic acid) and strongly embryotoxic (hydroxyurea) as listed by European Centre for the Validation of Alternative Methods (ECVAM) and examined endpoints of cell viability and neuronal protein marker expression specific to the central nervous system, to identify developmental neurotoxins. Following induction of neuronal differentiation, valproic acid had the most significant effect on neurogenesis, in terms of reduced viability and decreased neuronal markers. Lithium had least effect on viability and did not significantly alter the expression of neuronal markers. Hydroxyurea significantly reduced cell viability but did not affect neuronal protein marker expression. Acrylamide reduced neurosphere viability but did not affect neuronal protein marker expression. Overall, this NT2/D1-based neurosphere model of neurogenesis, may provide the basis for a model of developmental neurotoxicity in vitro

  5. Operating Procedures to Improve Efficiencies of In vitro Exposure Systems at the Air-Liquid Interface

    Science.gov (United States)

    The expanding use of in vitro exposure systems for toxicity assessments has created regulatory concerns. Many of these same concerns surround the proper conduct of in vivo inhalation toxicology studies that are addressed in guidelines and Good Laboratory Practice (GLPs) regulatio...

  6. In vitro human digestion test to monitor the dissolution of silver nanoparticles

    International Nuclear Information System (INIS)

    Bove, P; Sabella, S; Malvindi, M A

    2017-01-01

    Nanotechnology is a scientific revolution that the food industry has experienced over the last years. Widely employed as food additives and/or food contact materials in consumer products, silver nanoparticles are an example of this innovation. However, their increasing use makes also likely the human ingestion, thus requiring a proper risk analysis. In this framework, a comprehensive characterization of biotransformation of silver nanoparticles in biological fluids is fundamental for the regulatory needs. Herein, we aimed at studying the dissolution behaviour of silver nanoparticles using an in vitro test, which simulates the human oral ingestion of NPs during their passage through the gastrointestinal tract. The nanoparticle suspensions were characterized in the different digestion phases using several techniques to follow the changes of key physical properties ( e.g. , size, surface charge and plasmon peak) and to quantify the biotransformed products arisen by the process, as for example free silver ions. (paper)

  7. In vitro testing of defense reactions in zygotic and somatic embryos of Abies numidica

    Directory of Open Access Journals (Sweden)

    Jiří Hřib

    2011-01-01

    Full Text Available Defense of desiccated cotyledonary somatic embryos and mature zygotic embryos of Abies numidica was tested in vitro by dual cultures with tester, fungus Phaeolus schweinitzii. Both types of embryos expressed defense reactions manifested by inhibited growth of fungal tester towards the embryos. Mycelial growth was described by logistic sigmoid growth model with a single asymptote. Mutual comparisons of mycelial growth in presence of zygotic and somatic embryos showed significant differences in parameters of mycelium growth curves towards the embryos. Larger defense reactions were observed in zygotic embryos relative to somatic embryos and unlimited control cultivations without embryo. The possible role of auxin in the defense response of plant embryos is discussed.

  8. In vitro toxicity test of nano-sized magnesium oxide synthesized via solid-phase transformation

    Science.gov (United States)

    Zheng, Jun; Zhou, Wei

    2018-04-01

    Nano-sized magnesium oxide (MgO) has been a promising potential material for biomedical pharmaceuticals. In the present investigation, MgO nanoparticles synthesized through in-situ solid-phase transformation based on the previous work (nano-Mg(OH)2 prepared by precipitation technique) using magnesium nitrate and sodium hydroxide. The phase structure and morphology of the MgO nanoparticles are characterized by X-ray powder diffraction (XRD), selected area electronic diffraction (SAED) and transmission electron microscopy (TEM) respectively. In vitro hemolysis tests are adopted to evaluate the toxicity of the synthesized nano-MgO. The results evident that nano-MgO with lower concentration is slightly hemolytic, and with concentration increasing nano-MgO exhibit dose-responsive hemolysis.

  9. Prenatal developmental toxicity testing of petroleum substances: Application of the mouse embryonic stem cell test (EST) to compare in vitro potencies with potencies observed in vivo.

    Science.gov (United States)

    Kamelia, Lenny; Louisse, Jochem; de Haan, Laura; Rietjens, Ivonne M C M; Boogaard, Peter J

    2017-10-01

    Prenatal developmental toxicity (PDT) as observed with some petroleum substances (PS) has been associated with the presence of 3-7 ring polycyclic aromatic hydrocarbons (PAHs). In the present study, the applicability of ES-D3 cell differentiation assay of the EST to evaluate in vitro embryotoxicity potencies of PS and gas-to-liquid (GTL) products as compared to their in vivo potencies was investigated. DMSO-extracts of a range of PS, containing different amounts of PAHs, and GTL-products, which are devoid of PAHs, were tested in the ES-D3 cell proliferation and differentiation assays of the EST. The results show that PS inhibited the differentiation of ES-D3 cells into cardiomyocytes in a concentration-dependent manner at non-cytotoxic concentrations, and that their potency was proportional to their PAH content. In contrast, as expected, GTL-products did not inhibit ES-D3 cell viability or differentiation at all. The in vitro PDT potencies were compared to published in vivo PDT studies, and a good correlation was found between in vitro and in vivo results (R 2 =0.97). To conclude, our results support the hypothesis that PAHs are the primary inducers of the PDT in PS. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

  10. On the Road to Development of an in Vitro Permeation Test (IVPT) Model to Compare Heat Effects on Transdermal Delivery Systems: Exploratory Studies with Nicotine and Fentanyl.

    Science.gov (United States)

    Shin, Soo Hyeon; Ghosh, Priyanka; Newman, Bryan; Hammell, Dana C; Raney, Sam G; Hassan, Hazem E; Stinchcomb, Audra L

    2017-09-01

    At elevated temperatures, the rate of drug release and skin permeation from transdermal delivery systems (TDS) may be higher than at a normal skin temperature. The aim of this study was to compare the effect of heat on the transdermal delivery of two model drugs, nicotine and fentanyl, from matrix-type TDSs with different formulations, using in vitro permeation tests (IVPT). IVPT experiments using pig skin were performed on two nicotine and three fentanyl TDSs. Both continuous and transient heat exposures were investigated by applying heat either for the maximum recommended TDS wear duration or for short duration. Continuous heat exposure for the two nicotine TDSs resulted in different effects, showing a prolonged heat effect for one product but not the other. The J max enhancement ratio due to the continuous heat effect was comparable between the two nicotine TDS, but significantly different (p drug from the skin depot after TDS removal differently for two drugs, with fentanyl exhibiting a longer heat effect. This exploratory work suggests that an IVPT study may be able to discriminate differences in transdermal drug delivery when different TDS are exposed to elevated temperatures. However, the clinical significance of IVPT heat effects studies should be further explored by conducting in vivo clinical studies with similar study designs.

  11. Light Duty Utility Arm system pre-operational (cold test) test plan

    International Nuclear Information System (INIS)

    Bennett, K.L.

    1995-01-01

    The Light Duty Utility (LDUA) Cold Test Facility, located in the Hanford 400 Area, will be used to support cold testing (pre- operational tests) of LDUA subsystems. Pre-operational testing is composed of subsystem development testing and rework activities, and integrated system qualification testing. Qualification testing will be conducted once development work is complete and documentation is under configuration control. Operational (hot) testing of the LDUA system will follow the testing covered in this plan and will be covered in a separate test plan

  12. Phytochemical Composition and Antioxidant Potential of Ruta graveolens L. In Vitro Culture Lines

    Directory of Open Access Journals (Sweden)

    Renuka Diwan

    2012-01-01

    Full Text Available Ruta graveolens L. is a medicinal plant used in traditional systems of medicine for treatment of psoriasis, vitiligo, leucoderma, and lymphomas with well-known anti-inflammatory and anticancer properties. Therefore antioxidant potential of R. graveolens (in planta and in vitro was investigated. As antioxidants present in plant extracts are multifunctional, their activity and mechanism depends on the composition and conditions of the test system. Therefore, the total antioxidant capacity was evaluated using assays that detect different antioxidants: free radical scavenging (DPPH and ABTS, transition metal ion reduction (phosphomolybdenum assay, reducing power, and nitric oxide reduction. Content of furanocoumarin-bergapten in the extracts showed good corelation with free radical scavenging, transition metal reduction and reducing power, while total phenolic content showed good corelation with nitric oxide reduction potential. Antioxidant activity of in vitro cultures was significantly higher compared to in vivo plant material. The present study is the first report on comprehensive study of antioxidant activity of R. graveolens and its in vitro cultures.

  13. 2-Aminoanthracene, 5-fluorouracil, colchicine, benzo[a]pyrene, cadmium chloride and cytosine arabinoside tested in the in vitro mammalian cell micronucleus test (MNvit) in Chinese hamster ovary (CHO) cells at Covance Laboratories, Harrogate UK in support of OECD draft Test Guideline 487.

    Science.gov (United States)

    Whitwell, James; Fowler, Paul; Allars, Sarah; Jenner, Karen; Lloyd, Melvyn; Wood, Debbie; Smith, Katie; Young, Jamie; Jeffrey, Laura; Kirkland, David

    2010-10-29

    The reference genotoxic agents 2-aminoanthracene (a metabolism dependent weak clastogen), 5-fluorouracil (a nucleoside analogue, characterised by a steep dose response profile), colchicine (an aneugen that inhibits tubulin polymerisation), benzo[a]pyrene (a polycyclic aromatic hydrocarbon requiring metabolic activation), cadmium chloride (an inorganic carcinogen), and cytosine arabinoside (a nucleoside analogue that inhibits the gap-filling step of excision repair) were tested in the in vitro micronucleus assay using the Chinese hamster ovary (CHO) cell line at Covance Laboratories, Harrogate, UK. All chemicals were treated in the absence and presence of cytokinesis block (via addition of cytochalasin B) with this work forming part of a collaborative evaluation of the toxicity measures recommended in the draft OECD Test Guideline 487 on the In vitro Mammalian Cell Micronucleus Test (MNvit). The toxicity measures used, detecting a possible combination of both cytostasis and cell death (though not cell death directly), were relative population doubling, relative increase in cell counts and relative cell counts for treatments in the absence of cytokinesis block, and replication index in the presence of cytokinesis block. All of the chemicals tested either gave marked positive increases in the percentage of micronucleated cells with and without cytokinesis block, or did not induce micronuclei at concentrations giving approximately 50-60% toxicity (cytostasis and cell death) or less by all of the toxicity measures used. The outcome from this series of tests supports the use of relative increase in cell counts and relative population doubling, as well as relative cell counts, as appropriate measures of cytotoxicity for the non-cytokinesis blocked in vitro micronucleus assay. Copyright © 2010 Elsevier B.V. All rights reserved.

  14. Testing on air cleaning systems: Testing of the components in-place tests

    International Nuclear Information System (INIS)

    Billard, F.; Brion, J.

    1967-01-01

    The reliability of air cleaning systems is dependent on testing they are submitted to. Although in-place tests are the most important as they act as final tests upon achieved plants, component tests are necessary too. They allow detection of defective units before they are installed, partition of unit defects from mounting defects and they are more sensitive. For similar reasons, material teats are most useful. The various tests are described, about aerosol filters for one part, iodine trap for the other. The checked features are: materials nature, units sizes, efficiency, air resistance, flammability, humidity resistance, temperature resistance, adsorbent friability, etc... On iodine trapping systems, small check traps, working by-pass with the main trap are periodically subjected to efficiency test. This control allow to cut down the in-place tests frequency, particularly when poisoning from organic vapours is to be feared. (authors) [fr

  15. In Vitro Developmental Toxicology Screens: A Report on the Progress of the Methodology and Future Applications.

    Science.gov (United States)

    Zhang, Cindy; Ball, Jonathan; Panzica-Kelly, Julie; Augustine-Rauch, Karen

    2016-04-18

    There has been increasing focus on generation and assessment of in vitro developmental toxicology models for assessing teratogenic liability of chemicals. The driver for this focus has been to find reliable in vitro assays that will reduce or replace the use of in vivo tests for assessing teratogenicity. Such efforts may be eventually applied in testing pharmaceutical agents where a developmental toxicology assay or battery of assays may be incorporated into regulatory testing to replace one of the two species currently used in teratogenic assessment. Such assays may be eventually applied in testing a broader spectrum of chemicals, supporting efforts aligned with Tox21 strategies and responding to REACH legislation. This review describes the developmental toxicology assays that are of focus in these assessments: rodent whole embryo culture, zebrafish embryo assays, and embryonic stem cell assays. Progress on assay development as well as future directions of how these assays are envisioned to be applied for broader safety testing of chemicals are discussed. Altogether, the developmental model systems described in this review provide rich biological systems that can be utilized in better understanding teratogenic mechanisms of action of chemotypes and are promising in providing proactive safety assessment related to developmental toxicity. Continual advancements in refining/optimizing these in vitro assays are anticipated to provide a robust data set to provide thoughtful assessment of how whole animal teratogenicity evaluations can be reduced/refined in the future.

  16. Gene expression data from acetaminophen-induced toxicity in human hepatic in vitro systems and clinical liver samples

    Directory of Open Access Journals (Sweden)

    Robim M. Rodrigues

    2016-06-01

    Full Text Available This data set is composed of transcriptomics analyses of (i liver samples from patients suffering from acetaminophen-induced acute liver failure (ALF and (ii hepatic cell systems exposed to acetaminophen and their respective controls. The in vitro systems include widely employed cell lines i.e. HepaRG and HepG2 cells as well as a novel stem cell-derived model i.e. human skin-precursors-derived hepatocyte-like cells (hSKP-HPC. Data from primary human hepatocytes was also added to the data set “Open TG-GATEs: a large-scale toxicogenomics database” (Igarashi et al., 2015 [1]. Changes in gene expression due to acetaminophen intoxication as well as comparative information between human in vivo and in vitro samples are provided. The microarray data have been deposited in NCBI׳s Gene Expression Omnibus and are accessible through GEO Series accession number GEO: GSE74000. The provided data is used to evaluate the predictive capacity of each hepatic in vitro system and can be directly compared with large-scale publically available toxicogenomics databases. Further interpretation and discussion of these data feature in the corresponding research article “Toxicogenomics-based prediction of acetaminophen-induced liver injury using human hepatic cell systems” (Rodrigues et al., 2016 [2].

  17. Safety testing for LHC access system

    CERN Document Server

    Valentini, F; Ninin, P; Scibile, S

    2008-01-01

    In the domain of Safety Real-Time Systems the problem of testing represents always a big effort in terms of time, costs and efficiency to guarantee an adequate coverage degree. Exhaustive tests may, in fact, not be practicable for large and distributed systems. This paper describes the testing process followed during the validation of the CERN's LHC Access System [1], responsible for monitoring and preventing physical risks for the personnel accessing the underground areas. In the paper we also present a novel strategy for the testing problem, intended to drastically reduce the time for the test patterns generation and execution. In particular, we propose a methodology for blackbox testing that relies on the application of Model Checking techniques. Model Checking is a formal method from computer science, commonly adopted to prove correctness of system’s models through an automatic system’s state space exploration against some property formulas.

  18. Extrapolation of systemic bioavailability assessing skin absorption and epidermal and hepatic metabolism of aromatic amine hair dyes in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Manwaring, John, E-mail: manwaring.jd@pg.com [Procter & Gamble Inc., Mason Business Center, Mason, OH 45040 (United States); Rothe, Helga [Procter & Gamble Service GmbH, Sulzbacher Str. 40, 65823 Schwalbach am Taunus (Germany); Obringer, Cindy; Foltz, David J.; Baker, Timothy R.; Troutman, John A. [Procter & Gamble Inc., Mason Business Center, Mason, OH 45040 (United States); Hewitt, Nicola J. [SWS, Erzhausen (Germany); Goebel, Carsten [Procter & Gamble Service GmbH, Sulzbacher Str. 40, 65823 Schwalbach am Taunus (Germany)

    2015-09-01

    Approaches to assess the role of absorption, metabolism and excretion of cosmetic ingredients that are based on the integration of different in vitro data are important for their safety assessment, specifically as it offers an opportunity to refine that safety assessment. In order to estimate systemic exposure (AUC) to aromatic amine hair dyes following typical product application conditions, skin penetration and epidermal and systemic metabolic conversion of the parent compound was assessed in human skin explants and human keratinocyte (HaCaT) and hepatocyte cultures. To estimate the amount of the aromatic amine that can reach the general circulation unchanged after passage through the skin the following toxicokinetically relevant parameters were applied: a) Michaelis–Menten kinetics to quantify the epidermal metabolism; b) the estimated keratinocyte cell abundance in the viable epidermis; c) the skin penetration rate; d) the calculated Mean Residence Time in the viable epidermis; e) the viable epidermis thickness and f) the skin permeability coefficient. In a next step, in vitro hepatocyte K{sub m} and V{sub max} values and whole liver mass and cell abundance were used to calculate the scaled intrinsic clearance, which was combined with liver blood flow and fraction of compound unbound in the blood to give hepatic clearance. The systemic exposure in the general circulation (AUC) was extrapolated using internal dose and hepatic clearance, and C{sub max} was extrapolated (conservative overestimation) using internal dose and volume of distribution, indicating that appropriate toxicokinetic information can be generated based solely on in vitro data. For the hair dye, p-phenylenediamine, these data were found to be in the same order of magnitude as those published for human volunteers. - Highlights: • An entirely in silico/in vitro approach to predict in vivo exposure to dermally applied hair dyes • Skin penetration and epidermal conversion assessed in human

  19. Extrapolation of systemic bioavailability assessing skin absorption and epidermal and hepatic metabolism of aromatic amine hair dyes in vitro

    International Nuclear Information System (INIS)

    Manwaring, John; Rothe, Helga; Obringer, Cindy; Foltz, David J.; Baker, Timothy R.; Troutman, John A.; Hewitt, Nicola J.; Goebel, Carsten

    2015-01-01

    Approaches to assess the role of absorption, metabolism and excretion of cosmetic ingredients that are based on the integration of different in vitro data are important for their safety assessment, specifically as it offers an opportunity to refine that safety assessment. In order to estimate systemic exposure (AUC) to aromatic amine hair dyes following typical product application conditions, skin penetration and epidermal and systemic metabolic conversion of the parent compound was assessed in human skin explants and human keratinocyte (HaCaT) and hepatocyte cultures. To estimate the amount of the aromatic amine that can reach the general circulation unchanged after passage through the skin the following toxicokinetically relevant parameters were applied: a) Michaelis–Menten kinetics to quantify the epidermal metabolism; b) the estimated keratinocyte cell abundance in the viable epidermis; c) the skin penetration rate; d) the calculated Mean Residence Time in the viable epidermis; e) the viable epidermis thickness and f) the skin permeability coefficient. In a next step, in vitro hepatocyte K m and V max values and whole liver mass and cell abundance were used to calculate the scaled intrinsic clearance, which was combined with liver blood flow and fraction of compound unbound in the blood to give hepatic clearance. The systemic exposure in the general circulation (AUC) was extrapolated using internal dose and hepatic clearance, and C max was extrapolated (conservative overestimation) using internal dose and volume of distribution, indicating that appropriate toxicokinetic information can be generated based solely on in vitro data. For the hair dye, p-phenylenediamine, these data were found to be in the same order of magnitude as those published for human volunteers. - Highlights: • An entirely in silico/in vitro approach to predict in vivo exposure to dermally applied hair dyes • Skin penetration and epidermal conversion assessed in human skin explants and

  20. A surprisingly poor correlation between in vitro and in vivo testing of biomaterials for bone regeneration : results of a multicentre analysis

    NARCIS (Netherlands)

    Hulsart-Billström, G.; Dawson, J.I.; Hofmann, S.; Müller, R.; Stoddart, M.J.; Alini, M.; Redl, H.; El Haj, A.; Brown, R.; Salih, V.; Hilborn, J.; Larsson, S.; Oreffo, R.O.C.

    2016-01-01

    New regenerative materials and approaches need to be assessed through reliable and comparable methods for rapid translation to the clinic. There is a considerable need for proven in vitro assays that are able to reduce the burden on animal testing, by allowing assessment of biomaterial utility

  1. Evaluation of genotoxicity of nitrile fragrance ingredients using in vitro and in vivo assays.

    Science.gov (United States)

    Bhatia, S P; Politano, V T; Api, A M

    2013-09-01

    Genotoxicity studies were conducted on a group of 8 fragrance ingredients that belong to the nitrile family. These nitriles are widely used in consumer products however there is very limited data in the literature regarding the genotoxicity of these nitriles. The 8 nitriles were assessed for genotoxicity using an Ames test, in vitro chromosome aberration test or in vitro micronucleus test. The positive results observed in the in vitro tests were further investigated using an in vivo micronucleus test. The results from these different tests were compared and these 8 nitriles are not considered to be genotoxic. Dodecanitrile and 2,2,3-trimethylcyclopent-3-enylacetonitrile were negative in the in vitro chromosome aberration test and in vitro micronucleus test, respectively. While citronellyl nitrile, 3-methyl-5-phenylpentanenitrile, cinnamyl nitrile, and 3-methyl-5-phenylpent-2-enenitrile revealed positive results in the in vitro tests, but confirmatory in vivo tests determined these nitriles to be negative in the in vivo micronucleus assay. The remaining two nitriles (benzonitrile and α-cyclohexylidene benzeneacetonitrile) were negative in the in vivo micronucleus test. This study aims to evaluate the genotoxicity potential of these nitriles as well as enrich the literature with genotoxicity data on fragrance ingredients. Copyright © 2013 Elsevier Ltd. All rights reserved.

  2. N-terminally truncated GADD34 proteins are convenient translation enhancers in a human cell-derived in vitro protein synthesis system.

    Science.gov (United States)

    Mikami, Satoshi; Kobayashi, Tominari; Machida, Kodai; Masutani, Mamiko; Yokoyama, Shigeyuki; Imataka, Hiroaki

    2010-07-01

    Human cell-derived in vitro protein synthesis systems are useful for the production of recombinant proteins. Productivity can be increased by supplementation with GADD34, a protein that is difficult to express in and purify from E. coli. Deletion of the N-terminal 120 or 240 amino acids of GADD34 improves recovery of this protein from E. coli without compromising its ability to boost protein synthesis in an in vitro protein synthesis system. The use of N-terminally truncated GADD34 proteins in place of full-length GADD34 should improve the utility of human cell-based cell-free protein synthesis systems.

  3. SP-100 nuclear assembly test: Test assembly functional requirements and system arrangement

    International Nuclear Information System (INIS)

    Fallas, T.T.; Gluck, R.; Motwani, K.; Clay, H.; O'Neill, G.

    1991-01-01

    This paper describes the functional requirements and the system that will be tested to validate the reactor, flight shield, and flight controller of the SP-100 Generic Flight System (GFS). The Nuclear Assembly Test (NAT) consists of the test article (SP-100 reactor with control devices and the flight shield) and its supporting systems. The NAT test assembly is being designed by GE. Westinghouse Hanford Company (WHC) is designing the test cell and vacuum vessel system that will contain the NAT test assembly (Renkey et al. 1989). Preliminary design reviews have been completed and the final design is under way

  4. CMIF ECLS system test findings

    Science.gov (United States)

    Schunk, Richard G.; Carrasquillo, Robyn L.; Ogle, Kathyrn Y.; Wieland, Paul O.; Bagdigian, Robert M.

    1989-01-01

    During 1987 three Space Station integrated Environmental Control and Life Support System (ECLSS) tests were conducted at the Marshall Space Flight Center (MSFC) Core Module Integration Facility (CMIF) as part of the MSFC ECLSS Phase II test program. The three tests ranged in duration from 50 to 150 hours and were conducted inside of the CMIF module simulator. The Phase II partial integrated system test configuration consisted of four regenerative air revitalization subsystems and one regenerative water reclamation subsystem. This paper contains a discussion of results and lessons learned from the Phase II test program. The design of the Phase II test configuration and improvements made throughout the program are detailed. Future plans for the MSFC CMIF test program are provided, including an overview of planned improvements for the Phase III program.

  5. High Power Alternator Test Unit (ATU) Electrical System Test

    Science.gov (United States)

    Birchenough, Arthur; Hervol, David

    2007-01-01

    The Alternator Test Unit (ATU) in the Lunar Power System Facility (LPSF) located at the NASA Glenn Research Center (GRC) in Cleveland, OH was used to simulate the operating conditions and evaluate the performance of the ATU and it s interaction with various LPSF components in accordance with the JIMO AC Power System Requirements. The testing was carried out at the breadboard development level. Results of these tests will be used for the development and validation of analytical models for performance and lifetime prediction.

  6. In vitro testing to diagnose venom allergy and monitor immunotherapy: a placebo-controlled, crossover trial.

    Science.gov (United States)

    Brown, S G A; Haas, M A; Black, J A; Parameswaran, A; Woods, G M; Heddle, R J

    2004-05-01

    In people with a history of sting allergy, only prior reaction severity and older age are known to predict subsequent reaction risk. Furthermore, no diagnostic test other than a deliberate sting challenge has been found to identify people in whom venom immunotherapy (VIT) has been unsuccessful. We aimed to assess the utility of a number of in vitro tests to diagnose venom allergy and to monitor immunotherapy. During a double-blind randomized placebo-controlled crossover trial of Myrmecia pilosula ant VIT the following venom-specific tests were performed at enrolment, and at completion of treatment prior to a diagnostic sting challenge; leucocyte stimulation index (SI), IL-4 production, IgE RAST, histamine release test (HRT), leukotriene release test (LRT) and basophil activation test (BAT). Intradermal venom skin testing (VST) was also performed at trial entry. Only VST and HRT identified those at risk of sting anaphylaxis in the placebo group. Although IgE RAST, leucocyte SI and IL-4 production, LRT and BAT all correlated well with intradermal VSTs, they did not predict sting challenge outcome. After successful VIT, venom-induced leucocyte IL-4 production tended to fall, whereas IgE RAST increased and a natural decline in HRT reactivity was reversed. A confounding seasonal affect on laboratory results was suspected. The HRT warrants further assessment for diagnosis of venom allergy. Uninformative performance of the commercially available LRT and BAT tests may be due to pre-incubation with IL-3. None of the tests evaluated appear to be reliable markers of successful VIT.

  7. An in vitro approach for comparative interspecies metabolism of agrochemicals.

    Science.gov (United States)

    Whalley, Paul M; Bartels, Michael; Bentley, Karin S; Corvaro, Marco; Funk, Dorothee; Himmelstein, Matthew W; Neumann, Birgit; Strupp, Christian; Zhang, Fagen; Mehta, Jyotigna

    2017-08-01

    The metabolism and elimination of a xenobiotic has a direct bearing on its potential to cause toxicity in an organism. The confidence with which data from safety studies can be extrapolated to humans depends, among other factors, upon knowing whether humans are systemically exposed to the same chemical entities (i.e. a parent compound and its metabolites) as the laboratory animals used to study toxicity. Ideally, to understand a metabolite in terms of safety, both the chemical structure and the systemic exposure would need to be determined. However, as systemic exposure data (i.e. blood concentration/time data of test material or metabolites) in humans will not be available for agrochemicals, an in vitro approach must be taken. This paper outlines an in vitro experimental approach for evaluating interspecies metabolic comparisons between humans and animal species used in safety studies. The aim is to ensure, where possible, that all potential human metabolites are also present in the species used in the safety studies. If a metabolite is only observed in human in vitro samples and is not present in a metabolic pathway defined in the toxicological species already, the toxicological relevance of this metabolite must be evaluated. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

  8. ENRAIZAMENTO DE CRAVO (Dianthus caryophyllus L. IN VITRO E EX VITRO

    Directory of Open Access Journals (Sweden)

    G.R.F CUZZUOL

    1996-01-01

    Full Text Available Plântulas de cravo (Dianthus caryophyllus micropropagadas durante várias gerações pelo período de um ano, foram enraizadas in vitro com AIA, ANA e AIB nas concentrações de 0,0; 0,25; 0,5 e 1,0 mg/l, em fatorial do tipo 3 x 4, com todos os tratamentos promovendo a formação de raízes, mas não diferindo do controle. Foi confrontado em condição autotrófica, o desempenho entre plântulas enraizadas in vitro na presença e ausência do regulador AIA 0,5 mg/l e plântulas enraizadas ex vitro, sem nenhuma diferença quanto ao comprimento da parte aérea. Para a variável produção de massa de matéria seca os melhores resultados foram proporcionados pelas plantas que passaram pela fase de enraizamento in vitro, tendo o sistema radicular efeito sinergístico no crescimento da parte aérea.Plantlets of carnation (Dianthus caryophyllus L. micropropagated through several generations during one year, were observed with respect to rooting in vitro, in the presence of IAA , NAA and IBA, at the following concentrations: 0,0; 0,25; 0,5 and 1,0 mg/l. All treatments promoted root formation, however no differences were detected in comparison to control. As far as the lenght of the aerial part is concerned no difference was observed between in vitro rooting. in the presence or absence of IAA 0,5 mg/l, and ex vitro rooting. Plantlets which were rooted in vitro conditions showed higher production of fresh matter then those rooted ex vitro. The root system had a synergistic effect on the growth of the aerial part.

  9. Test Telemetry And Command System (TTACS)

    Science.gov (United States)

    Fogel, Alvin J.

    1994-01-01

    The Jet Propulsion Laboratory has developed a multimission Test Telemetry and Command System (TTACS) which provides a multimission telemetry and command data system in a spacecraft test environment. TTACS reuses, in the spacecraft test environment, components of the same data system used for flight operations; no new software is developed for the spacecraft test environment. Additionally, the TTACS is transportable to any spacecraft test site, including the launch site. The TTACS is currently operational in the Galileo spacecraft testbed; it is also being provided to support the Cassini and Mars Surveyor Program projects. Minimal personnel data system training is required in the transition from pre-launch spacecraft test to post-launch flight operations since test personnel are already familiar with the data system's operation. Additionally, data system components, e.g. data display, can be reused to support spacecraft software development; and the same data system components will again be reused during the spacecraft integration and system test phases. TTACS usage also results in early availability of spacecraft data to data system development and, as a result, early data system development feedback to spacecraft system developers. The TTACS consists of a multimission spacecraft support equipment interface and components of the multimission telemetry and command software adapted for a specific project. The TTACS interfaces to the spacecraft, e.g., Command Data System (CDS), support equipment. The TTACS telemetry interface to the CDS support equipment performs serial (RS-422)-to-ethernet conversion at rates between 1 bps and 1 mbps, telemetry data blocking and header generation, guaranteed data transmission to the telemetry data system, and graphical downlink routing summary and control. The TTACS command interface to the CDS support equipment is nominally a command file transferred in non-real-time via ethernet. The CDS support equipment is responsible for

  10. Vitrification Facility integrated system performance testing report

    International Nuclear Information System (INIS)

    Elliott, D.

    1997-01-01

    This report provides a summary of component and system performance testing associated with the Vitrification Facility (VF) following construction turnover. The VF at the West Valley Demonstration Project (WVDP) was designed to convert stored radioactive waste into a stable glass form for eventual disposal in a federal repository. Following an initial Functional and Checkout Testing of Systems (FACTS) Program and subsequent conversion of test stand equipment into the final VF, a testing program was executed to demonstrate successful performance of the components, subsystems, and systems that make up the vitrification process. Systems were started up and brought on line as construction was completed, until integrated system operation could be demonstrated to produce borosilicate glass using nonradioactive waste simulant. Integrated system testing and operation culminated with a successful Operational Readiness Review (ORR) and Department of Energy (DOE) approval to initiate vitrification of high-level waste (HLW) on June 19, 1996. Performance and integrated operational test runs conducted during the test program provided a means for critical examination, observation, and evaluation of the vitrification system. Test data taken for each Test Instruction Procedure (TIP) was used to evaluate component performance against system design and acceptance criteria, while test observations were used to correct, modify, or improve system operation. This process was critical in establishing operating conditions for the entire vitrification process

  11. Bacterial challenge of NISSHO ultrafilter ETF 609: results of in vitro testing.

    Science.gov (United States)

    Krautzig, S; Lonnemann, G; Shaldon, S; Koch, K M

    1996-07-01

    In hemodialysis, a certain degree of bacterial contamination on the dialysate side is a regular finding. Concern has been growing that this contamination may lead to a chronic inflammatory response in the patient. Ultrafiltration of dialysate can be used to reduce bacterial content and levels of cytokine-inducing substances upstream of the patient's dialyzer. The aim of this study was to test in vitro the rejection capacity of a polysulfone hollow-fiber ultrafilter (ETF 609, NISSHO Co., Osaka, Japan) challenged with bacterial filtrates derived from Pseudomonas aeruginosa PA103. Results showed a reduction of interleukin-1 beta-inducing activity (measured on peripheral blood mononuclear cells) from 5,035 +/- 394 pg/ml prefilter to nondetectable levels postfilter and endotoxin levels (limulus amebocyte lysate assay) of 4,167 +/- 1,079 versus 12 +/- 2 pg/ml, respectively. In conclusion, ultrafiltration of dialysate with the polysulfone ultrafilter ETF 609 leads to a potent reduction of cytokine-inducing activity.

  12. RESPONSE OF Cattleya forbesii ORCHID TO INCREASING SILICON CONCENTRATIONS IN VITRO

    Directory of Open Access Journals (Sweden)

    RONAN CARLOS COLOMBO

    2016-01-01

    Full Text Available Addition of Silicon (Si to culture media has been shown to improve the development of seedlings grown in vitro , and to reduce losses during the acclimatization phase. The objective of this study was to evaluate the in vitro growth of Cattleya forbesii (Orchidaceae in MS medium containing five different concentrations of SiO 2 (0.0, 0.5, 1.0, 1.5, and 2.0 g·L −1 . At day 200, the following variables were measured: number of roots, average length of the root system, leaf area, number of leaves and shoots, shoot height, fresh and dry masses of roots and shoots, water content of roots and shoots, and pH of the culture medium. Most variables decreased as the concentration of Si increased, reducing the in vitro vegetative growth of C. forbesii . Accumulation of Si in leaf tissues was detected by scanning electron microscopy, confirming uptake by plants. The Si source and concentrations tested showed no beneficial effect on in vitro growth of C. forbesii .

  13. 'Zipbody' leucine zipper-fused Fab in E. coli in vitro and in vivo expression systems.

    Science.gov (United States)

    Ojima-Kato, Teruyo; Fukui, Kansuke; Yamamoto, Hiroaki; Hashimura, Dai; Miyake, Shiro; Hirakawa, Yuki; Yamasaki, Tomomi; Kojima, Takaaki; Nakano, Hideo

    2016-04-01

    A small antibody fragment, fragment of antigen binding (Fab), is favorable for various immunological assays. However, production efficiency of active Fab in microorganisms depends considerably on the clones. In this study, leucine zipper-peptide pairs that dimerize in parallel (ACID-p1 (LZA)/BASE-p1 (LZB) or c-Jun/c-Fos) were fused to the C-terminus of heavy chain (Hc, VH-CH1) and light chain (Lc, VL-CL), respectively, to accelerate the association of Hc and Lc to form Fab in Escherichia coli in vivo and in vitro expression systems. The leucine zipper-fused Fab named 'Zipbody' was constructed using anti-E. coli O157 monoclonal antibody obtained from mouse hybridoma and produced in both in vitro and in vivo expression systems in an active form, whereas Fab without the leucine zipper fusion was not. Similarly, Zipbody of rabbit monoclonal antibody produced in in vitro expression showed significant activity. The purified, mouse Zipbody produced in the E. coli strain Shuffle T7 Express had specificity toward the antigen; in bio-layer interferometry analysis, the KD value was measured to be 1.5-2.0 × 10(-8) M. These results indicate that leucine zipper fusion to Fab C-termini markedly enhances active Fab formation in E. coli. © The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  14. Alginate based 3D hydrogels as an in vitro co-culture model platform for the toxicity screening of new chemical entities

    International Nuclear Information System (INIS)

    Lan, Shih-Feng; Starly, Binil

    2011-01-01

    Prediction of human response to potential therapeutic drugs is through conventional methods of in vitro cell culture assays and expensive in vivo animal testing. Alternatives to animal testing require sophisticated in vitro model systems that must replicate in vivo like function for reliable testing applications. Advancements in biomaterials have enabled the development of three-dimensional (3D) cell encapsulated hydrogels as in vitro drug screening tissue model systems. In this study, we have developed an in vitro platform to enable high density 3D culture of liver cells combined with a monolayer growth of target breast cancer cell line (MCF-7) in a static environment as a representative example of screening drug compounds for hepatotoxicity and drug efficacy. Alginate hydrogels encapsulated with serial cell densities of HepG2 cells (10 5 -10 8 cells/ml) are supported by a porous poly-carbonate disc platform and co-cultured with MCF-7 cells within standard cell culture plates during a 3 day study period. The clearance rates of drug transformation by HepG2 cells are measured using a coumarin based pro-drug. The platform was used to test for HepG2 cytotoxicity 50% (CT 50 ) using commercially available drugs which further correlated well with published in vivo LD 50 values. The developed test platform allowed us to evaluate drug dose concentrations to predict hepatotoxicity and its effect on the target cells. The in vitro 3D co-culture platform provides a scalable and flexible approach to test multiple-cell types in a hybrid setting within standard cell culture plates which may open up novel 3D in vitro culture techniques to screen new chemical entity compounds. - Graphical abstract: Display Omitted Highlights: → A porous support disc design to support the culture of desired cells in 3D hydrogels. → Demonstrated the co-culture of two cell types within standard cell-culture plates. → A scalable, low cost approach to toxicity screening involving multiple cell

  15. Testing in Vitro of an Apifitoterapeutic Formula Against Nosema spp.

    Directory of Open Access Journals (Sweden)

    Vasilica Savu

    2015-05-01

    Full Text Available Nosema, a parasitic disease that affects adult honey bees, has a directly correlation with the losses of bee colonies, until to depopulation. The target of our study was to determine the antinosema action of an apifitoterapeutic formula that was obtained in an earlier phase of our researches. In the present study, we have had two experiences (F and N formed by clinically healthy bees. The experimental bees have received, in vitro, naturally infested honey (7 spores by Nosema spp / field. The first experience (F, I-IX groups was treated with apifitoterapeutic formula (10 ml/ honey kg, for 10 days (from T1 to T2 moment, while the second experience (N, with X-XVIII groups was infested with naturally infested honey, for 20 days (from T1 to T2 moment. The first experience (F showed 22% positive diagnosed bees, while the second experience (N showed 89% positive diagnosed bees. In the first experience, the infestation degree was very weak (group I and weak (group III, while the other groups were negative. The antiparasitic formula has showed, in laboratory conditions, a positive impact on experimental honey bees, with an efficiency over 78%. In the further, testing prophylactically and therapeutically will be conducted on bee families.

  16. In vitro reactivity to concanavalin A of bovine lymphocytes after cryopreservation

    NARCIS (Netherlands)

    Kuil, H.

    1984-01-01

    Cryopreservation of bovine peripheral lymphocytes and its effect on the in vitro response to concanavalin A tested in a microculture system is described. Using DMSO as cryoprotectant in the medium, the cells were cooled to −30°C at 1.3°C/minute and further to −80°C at 6°C/minute and then rapidly to

  17. Neural differentiation of mouse embryonic stem cells as a tool to assess developmental neurotoxicity in vitro.

    Science.gov (United States)

    Visan, Anke; Hayess, Katrin; Sittner, Dana; Pohl, Elena E; Riebeling, Christian; Slawik, Birgitta; Gulich, Konrad; Oelgeschläger, Michael; Luch, Andreas; Seiler, Andrea E M

    2012-10-01

    Mouse embryonic stem cells (mESCs) represent an attractive cellular system for in vitro studies in developmental biology as well as toxicology because of their potential to differentiate into all fetal cell lineages. The present study aims to establish an in vitro system for developmental neurotoxicity testing employing mESCs. We developed a robust and reproducible protocol for fast and efficient differentiation of the mESC line D3 into neural cells, optimized with regard to chemical testing. Morphological examination and immunocytochemical staining confirmed the presence of different neural cell types, including neural progenitors, neurons, astrocytes, oligodendrocytes, and radial glial cells. Neurons derived from D3 cells expressed the synaptic proteins PSD95 and synaptophysin, and the neurotransmitters serotonin and γ-aminobutyric acid. Calcium ion imaging revealed the presence of functionally active glutamate and dopamine receptors. In addition, flow cytometry analysis of the neuron-specific marker protein MAP2 on day 12 after induction of differentiation demonstrated a concentration dependent effect of the neurodevelopmental toxicants methylmercury chloride, chlorpyrifos, and lead acetate on neuronal differentiation. The current study shows that D3 mESCs differentiate efficiently into neural cells involving a neurosphere-like state and that this system is suitable to detect adverse effects of neurodevelopmental toxicants. Therefore, we propose that the protocol for differentiation of mESCs into neural cells described here could constitute one component of an in vitro testing strategy for developmental neurotoxicity. Copyright © 2012 Elsevier Inc. All rights reserved.

  18. Test plan: Brayton Isotope Power System Ground Demonstration System (BIPS-GDS)

    International Nuclear Information System (INIS)

    1976-01-01

    The purpose of this test plan is to provide an overall outline of all testing to be accomplished on the GDS. Included in this test plan are administrative requirements, instrumentation accuracies, instrumentation, equipment definitions, system test setup, and facility installation. The test program will enable collection of sufficient data to establish material, component, and system design integrity. The data will also be used to establish and evaluate component and system performance and reliability characteristics, verification of proper system component integration prior to initiation of Phase II, and flight system (FS) development

  19. Continuous Improvement in Battery Testing at the NASA/JSC Energy System Test Area

    Science.gov (United States)

    Boyd, William; Cook, Joseph

    2003-01-01

    The Energy Systems Test Area (ESTA) at the Lyndon B. Johnson Space Center in Houston, Texas conducts development and qualification tests to fulfill Energy System Division responsibilities relevant to ASA programs and projects. EST A has historically called upon a variety of fluid, mechanical, electrical, environmental, and data system capabilities spread amongst five full-service facilities to test human and human supported spacecraft in the areas of propulsion systems, fluid systems, pyrotechnics, power generation, and power distribution and control systems. Improvements at ESTA are being made in full earnest of offering NASA project offices an option to choose a thorough test regime that is balanced with cost and schedule constraints. In order to continue testing of enabling power-related technologies utilized by the Energy System Division, an especially proactive effort has been made to increase the cost effectiveness and schedule responsiveness for battery testing. This paper describes the continuous improvement in battery testing at the Energy Systems Test Area being made through consolidation, streamlining, and standardization.

  20. In vitro efficacy and release study with anti-inflammatory drugs incorporated in adhesive transdermal drug delivery systems.

    Science.gov (United States)

    Meyer, Stefanie; Peters, Nils; Mann, Tobias; Wolber, Rainer; Pörtner, Ralf; Nierle, Jens

    2014-04-01

    The topical application of two different anti-inflammatory extracts incorporated in adhesive transdermal drug delivery systems (TDDSs) was investigated. Therefore, anti-inflammatory properties and percutaneous absorption behavior of adhesive TDDSs were characterized in vitro conducting experiments with a dermatologically relevant human skin model. Anti-inflammatory efficacy against UV irradiation of both TDDSs was determined in vitro with EpiDerm™. The reduction of the release of proinflammatory cytokines by topically applied TDDSs was compared with the reduction during the presence of the specific cyclooxygenase inhibitor diclofenac in the culture medium. A similar anti-inflammatory efficacy of the topically applied TDDSs in comparison with the use of diclofenac in the culture medium should be achieved. Furthermore, percutaneous absorption in efficacy tests was compared with percutaneous absorption in diffusion studies with porcine cadaver skin. Both the topically applied TDDSs showed a significant anti-inflammatory activity. Permeation coefficients through the stratum corneum and the epidermis gained from the release studies on porcine cadaver skin (Magnolia: 2.23·10(-5) cm/h, licorice: 4.68·10(-6) cm/h) were approximately five times lower than the permeation coefficients obtained with the EpiDerm™ skin model (Magnolia: 9.48·10(-5) cm/h, licorice: 24.0·10(-6) cm/h). Therefore, an adjustment of drug doses during experiments with the EpiDerm™ skin model because of weaker skin barrier properties should be considered.

  1. Testing Infrastructure for Operating System Kernel Development

    DEFF Research Database (Denmark)

    Walter, Maxwell; Karlsson, Sven

    2014-01-01

    Testing is an important part of system development, and to test effectively we require knowledge of the internal state of the system under test. Testing an operating system kernel is a challenge as it is the operating system that typically provides access to this internal state information. Multi......-core kernels pose an even greater challenge due to concurrency and their shared kernel state. In this paper, we present a testing framework that addresses these challenges by running the operating system in a virtual machine, and using virtual machine introspection to both communicate with the kernel...... and obtain information about the system. We have also developed an in-kernel testing API that we can use to develop a suite of unit tests in the kernel. We are using our framework for for the development of our own multi-core research kernel....

  2. Evaluation of Fatigue Behavior in Dental Implants from In Vitro Clinical Tests: A Systematic Review

    Directory of Open Access Journals (Sweden)

    Rosa Rojo

    2018-05-01

    Full Text Available In the area of dentistry, there is a wide variety of designs of dental implant and materials, especially titanium, which aims to avoid failures and increase their clinical durability. The purpose of this review was to evaluate fatigue behavior in different connections and implant materials, as well as their loading conditions and response to failure. In vitro tests under normal and dynamic loading conditions evaluating fatigue at implant and abutment connection were included. A search was conducted in PubMed, Scopus, and Science Direct. Data extraction was performed independently by two reviewers. The quality of selected studies was assessed using the Cochrane Handbook proposed by the tool for clinical trials. Nineteen studies were included. Fourteen studies had an unclear risk and five had high risk of bias. Due to the heterogeneity of the data and the evaluation of the quality of the studies, meta-analysis could not be performed. Evidence from this study suggests that both internal and morse taper connections presented a better behavior to failure. However, it is necessary to unify criteria in the methodological design of in vitro studies, following methodological guidelines and establishing conditions that allow the homogenization of designs in ISO (International Organization for Standardization standards.

  3. In vitro evaluation of matrix metalloproteinases as predictive testing for nickel, a model sensitizing agent

    International Nuclear Information System (INIS)

    Lamberti, Monica; Perfetto, Brunella; Costabile, Teresa; Canozo, Nunzia; Baroni, Adone; Liotti, Francesco; Sannolo, Nicola; Giuliano, Mariateresa

    2004-01-01

    The identification of potential damage due to chemical exposure in the workplace is a major health and regulatory concern. Traditional tests that measure both sensitization and elicitation responses require the use of animals. An alternative to this widespread use of experimental animals could have a crucial impact on risk assessment, especially for the preliminary screening of new molecules. We developed an in vitro model for the screening of potential toxic compounds. Human keratinocytes (HaCat) were used as target cells while matrix metalloproteinases (MMP) were selected as responders because they are key enzymes involved in extracellular matrix (ECM) degradation in physiological and pathological conditions. Chemical exposure was performed using nickel sulphate as a positive tester. Nickel contact induced upregulation of MMP-2 and IL-8 mRNA production. Molecular activation occurred even at very low nickel concentrations even though no phenotypic changes were observed. MMP-9 accumulation was found in the medium of treated cells with respect to controls. These observations led to the hypothesis that even minimal exposure can accumulate transcriptional activity resulting in long-term clinical signs after contact. Our simple in vitro model can be applied as a useful preliminary complement to the animal studies to screen the effects of new potential toxic compounds

  4. Tungsten carbide-cobalt as a nanoparticulate reference positive control in in vitro genotoxicity assays.

    Science.gov (United States)

    Moche, Hélène; Chevalier, Dany; Barois, Nicolas; Lorge, Elisabeth; Claude, Nancy; Nesslany, Fabrice

    2014-01-01

    With the increasing human exposure to nanoparticles (NP), the evaluation of their genotoxic potential is of significant importance. However, relevance for NP of the routinely used in vitro genotoxicity assays is often questioned, and a nanoparticulate reference positive control would therefore constitute an important step to a better testing of NP, ensuring that test systems are really appropriate. In this study, we investigated the possibility of using tungsten carbide-cobalt (WC-Co) NP as reference positive control in in vitro genotoxicity assays, including 2 regulatory assays, the mouse lymphoma assay and the micronucleus assay, and in the Comet assay, recommended for the toxicological evaluation of nanomedicines by the French Agency of Human Health Products (Afssaps). Through these assays, we were able to study different genetic endpoints in 2 cell types commonly used in regulatory genotoxicity assays: the L5178Y mouse lymphoma cell line and primary cultures of human lymphocytes. Our results showed that the use of WC-Co NP as positive control in in vitro genotoxicity assays was conceivable, but that different parameters have to be considered, such as cell type and treatment schedule. L5178Y mouse lymphoma cells did not provide satisfactory results in the 3 performed tests. However, human lymphocytes were more sensitive to genotoxic effects induced by WC-Co NP, particularly after a 24-h treatment in the in vitro micronucleus assay and after a 4-h treatment in the in vitro Comet assay. Under such conditions, WC-Co could be used as a nanoparticulate reference positive control in these assays.

  5. Darwin's concepts in a test tube: parallels between organismal and in vitro evolution.

    Science.gov (United States)

    Díaz Arenas, Carolina; Lehman, Niles

    2009-02-01

    The evolutionary process as imagined by Darwin 150 years ago is evident not only in nature but also in the manner in which naked nucleic acids and proteins experience the "survival of the fittest" in the test tube during in vitro evolution. This review highlights some of the most apparent evolutionary patterns, such as directional selection, purifying selection, disruptive selection, and iterative evolution (recurrence), and draws parallels between what happens in the wild with whole organisms and what happens in the lab with molecules. Advances in molecular selection techniques, particularly with catalytic RNAs and DNAs, have accelerated in the last 20 years to the point where soon any sort of complex differential hereditary event that one can ascribe to natural populations will be observable in molecular populations, and exploitation of these events can even lead to practical applications in some cases.

  6. Development of Final Running Test System for Digital Systems

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Kwang-Dae; Lee, Eui-Jong; Lim, Hee-Taek; Kim, Min-Seok [KHNP Central Research Institute, Daejeon (Korea, Republic of)

    2016-10-15

    In nuclear industry, the newly designed systems to upgrade are qualified to meet IEEE standards and the regulatory guidelines for their functions, performance and reliability requirements. Failure Mode and Effect Analysis, Fault Tree Analysis, and Hazard Analysis have been used to improve the reliability of the control system. To ensure the completeness of the software, the verification and validation processes are carried out during the development process. In spite of the many efforts depending on the analysis and procedures, there are limitations to improve the reliability. The lessons learned from the currently installed system failures show the incompleteness of the final integration test. The current point-to-point and logic-to-logic separate test procedures manually performed by the engineers can cause some procedures missed and have effects on the critical functions. The design processes of the digital systems are met in accordance with the international standards and regulatory guidelines. The lessons learned from the failures of the running digital systems showed the limitations of the current verification and validation efforts. The various improvements and attempts have been considered including the expert review processes and the completeness of the test. In this paper, the Final Running Test Method evaluating the completeness of the digital system using the control patterns and the Test System Architecture are proposed.

  7. Development of Final Running Test System for Digital Systems

    International Nuclear Information System (INIS)

    Lee, Kwang-Dae; Lee, Eui-Jong; Lim, Hee-Taek; Kim, Min-Seok

    2016-01-01

    In nuclear industry, the newly designed systems to upgrade are qualified to meet IEEE standards and the regulatory guidelines for their functions, performance and reliability requirements. Failure Mode and Effect Analysis, Fault Tree Analysis, and Hazard Analysis have been used to improve the reliability of the control system. To ensure the completeness of the software, the verification and validation processes are carried out during the development process. In spite of the many efforts depending on the analysis and procedures, there are limitations to improve the reliability. The lessons learned from the currently installed system failures show the incompleteness of the final integration test. The current point-to-point and logic-to-logic separate test procedures manually performed by the engineers can cause some procedures missed and have effects on the critical functions. The design processes of the digital systems are met in accordance with the international standards and regulatory guidelines. The lessons learned from the failures of the running digital systems showed the limitations of the current verification and validation efforts. The various improvements and attempts have been considered including the expert review processes and the completeness of the test. In this paper, the Final Running Test Method evaluating the completeness of the digital system using the control patterns and the Test System Architecture are proposed

  8. In Vitro Sensitivity Test in Antibiotics from the Fermentation Process in a Sugar-Alcohol Plant in the State of Paraná, Brazil

    OpenAIRE

    Lopes, Murilo Brandão; Faculdade de Apucarana – FAP; Silva, Thaís Medeiros Boldrin; Faculdade Metropolitana de Maringá – UNIFAMMA

    2011-01-01

    Since the development of different types of microorganisms is common during the fermentation process in sugar-alcohol plants, due to the processing states of prime matter, microbiological control is mandatory. In vitro sensitivity test is highly important for the fermentation process at sugar-alcohol plants since the type of antibiotic with the best antibacterial activity is evaluated. The test classifies antibiotics through their effects, namely, efficient, less efficient, slightly efficient...

  9. Rapid Analyses of Polyetheretherketone Wear Characteristics by Accelerated Wear Testing with Microfabricated Surfaces for Artificial Joint Systems.

    Science.gov (United States)

    Su, Chen-Ying; Kuo, Chien-Wei; Fang, Hsu-Wei

    2017-01-01

    Wear particle-induced biological responses are the major factors resulting in the loosening and then failure of total joint arthroplasties. It is feasible to improve the lubrication and reduce the wear of artificial joint system. Polyetheretherketone (PEEK) is considered as a potential bearing material due to its mechanical characteristics of resistance to fatigue strain. The PEEK wear particles have been indicated to be involved in biological responses in vitro, and further studies regarding the wear phenomena and wear particle generation are needed. In this study, we have established an accelerated wear testing system with microfabricated surfaces. Various contact pressures and lubricants have been utilized in the accelerated wear tests. Our results showed that increasing contact pressure resulted in an increase of wear particle sizes and wear rate, and the size of PEEK wear particles can be controlled by the feature size of microfabricated surfaces. These results provided the information rapidly about factors that affect the morphology and amount of PEEK wear particles and can be applied in the future for application of PEEK on the biological articulation system.

  10. A/T test system for production line; Seisan line yo A/T test system

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1999-03-10

    This paper introduces an A/T test system for production line made by Meidensha, Inc. Rates and technology of mounting auto-transmissions (A/T) in recent automobiles are improving year after year. A/T production line testers were fabricated and supplied, including those for European countries. This system has the following features: the system employs low inertia AC motor, and is capable of performing tests in close conditions to actual cars; an insulated gate bipolar transistor (IGBT) inverter was employed to achieve control accuracy improvement and noise reduction; high-velocity transient measurement is possible by using a computer system based on the WindowsNT; a solenoid current slant control was employed to respond to electronically controlled A/T; and noise and vibration shift feeling instrumentation can also be used on option. Number of supply is four sets of front engine/front wheel drive (FF) car A/T testers for overseas countries, and two sets of FF A/T tests for use in Japan. (NEDO)

  11. High Frequency Plant Regeneration System from Transverse Thin Cell Layer Section of In vitro Derived ‘Nadia’ Ginger Microrhizome

    Directory of Open Access Journals (Sweden)

    Dikash Singh THINGBAIJAM

    2014-03-01

    Full Text Available An efficient and reproducible procedure is outlined for rapid in vitro multiplication of Zingiber officinale var. ‘Nadia’ through high frequency shoot proliferation from transverse thin cell layer (tTCL sections of in vitro derived microrhizome. In vitro derived microrhizome of size 500 μm in thickness was used as initial explants for induction of somatic embryos. Among the different phytohormones tested, tTCL explants shows maximum calli proliferation in medium containing 2 mg/L 2,4-Dichlorophenoxyacetic acid (88.30±0.11%. Reduced concentration of 2,4 Dichlorophenoxyacetic acid was supplemented with different cytokinins for regeneration of callus. Among the different medium tested, optimum redifferentiation of somatic embryos were observed in medium containing 0.2 mg/L 2,4 Dichlorophenoxyacetic acid and 6.0 mg/L BAP (141.08±0.25. Clump of regenerated plantlets were further subculture and transfer into microrhizome inducing medium containing high sucrose concentration (8%. Plantlets with well developed microrhizome were successfully acclimatized and eventually transferred to the field. The application of studying embryo section for regeneration of plants might be useful alternative to ginger improvement programme. Histological analysis showed formation of somatic embryos and regenerated adventitious shoot.

  12. 21 CFR 862.3610 - Methamphetamine test system.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Methamphetamine test system. 862.3610 Section 862....3610 Methamphetamine test system. (a) Identification. A methamphetamine test system is a device intended to measure methamphetamine, a central nervous system stimulating drug, in serum, plasma, and urine...

  13. Launch Control System Software Development System Automation Testing

    Science.gov (United States)

    Hwang, Andrew

    2017-01-01

    The Spaceport Command and Control System (SCCS) is the National Aeronautics and Space Administration's (NASA) launch control system for the Orion capsule and Space Launch System, the next generation manned rocket currently in development. This system requires high quality testing that will measure and test the capabilities of the system. For the past two years, the Exploration and Operations Division at Kennedy Space Center (KSC) has assigned a group including interns and full-time engineers to develop automated tests to save the project time and money. The team worked on automating the testing process for the SCCS GUI that would use streamed simulated data from the testing servers to produce data, plots, statuses, etc. to the GUI. The software used to develop automated tests included an automated testing framework and an automation library. The automated testing framework has a tabular-style syntax, which means the functionality of a line of code must have the appropriate number of tabs for the line to function as intended. The header section contains either paths to custom resources or the names of libraries being used. The automation library contains functionality to automate anything that appears on a desired screen with the use of image recognition software to detect and control GUI components. The data section contains any data values strictly created for the current testing file. The body section holds the tests that are being run. The function section can include any number of functions that may be used by the current testing file or any other file that resources it. The resources and body section are required for all test files; the data and function sections can be left empty if the data values and functions being used are from a resourced library or another file. To help equip the automation team with better tools, the Project Lead of the Automated Testing Team, Jason Kapusta, assigned the task to install and train an optical character recognition (OCR

  14. In vitro-ex vivo correlations between a cell-laden hydrogel and mucosal tissue for screening composite delivery systems.

    Science.gov (United States)

    Blakney, Anna K; Little, Adam B; Jiang, Yonghou; Woodrow, Kim A

    2016-11-01

    Composite delivery systems where drugs are electrospun in different layers and vary the drug stacking-order are posited to affect bioavailability. We evaluated how the formulation characteristics of both burst- and sustained-release electrospun fibers containing three physicochemically diverse drugs: dapivirine (DPV), maraviroc (MVC) and tenofovir (TFV) affect in vitro and ex vivo release. We developed a poly(hydroxyethyl methacrylate) (pHEMA) hydrogel release platform for the rapid, inexpensive in vitro evaluation of burst- and sustained-release topical or dermal drug delivery systems with varying microarchitecture. We investigated properties of the hydrogel that could recapitulate ex vivo release into nonhuman primate vaginal tissue. Using a dimethyl sulfoxide extraction protocol and high-performance liquid chromatography analysis, we achieved >93% recovery from the hydrogels and >88% recovery from tissue explants for all three drugs. We found that DPV loading, but not stacking order (layers of fiber containing a single drug) or microarchitecture (layers with isolated drug compared to all drugs in the same layer) impacted the burst release in vitro and ex vivo. Our burst-release formulations showed a correlation for DPV accumulation between the hydrogel and tissue (R 2 =   0.80), but the correlation was not significant for MVC or TFV. For the sustained-release formulations, the PLGA/PCL content did not affect TFV release in vitro or ex vivo. Incorporation of cells into the hydrogel matrix improved the correlation between hydrogel and tissue explant release for TFV. We expect that this hydrogel-tissue mimic may be a promising preclinical model to evaluate topical or transdermal drug delivery systems with complex microarchitectures.

  15. Quality management systems for your in vitro fertilization clinic's laboratory: Why bother?

    Science.gov (United States)

    Olofsson, Jan I; Banker, Manish R; Sjoblom, Late Peter

    2013-01-01

    Several countries have in recent years introduced prescribed requirements for treatment and monitoring of outcomes, as well as a licensing or accreditation requirement for in vitro fertilization (IVF) clinics and their laboratories. It is commonplace for Assisted Reproductive Technology (ART) laboratories to be required to have a quality control system. However, more effective Total Quality Management systems are now being implemented by an increasing number of ART clinics. In India, it is now a requirement to have a quality management system in order to be accredited and to help meet customer demand for improved delivery of ART services. This review contains the proceedings a quality management session at the Indian Fertility Experts Meet (IFEM) 2010 and focuses on the creation of a patient-oriented best-in-class IVF laboratory.

  16. Experimental fast reactor JOYO MK-III functional test. Primary auxiliary cooling system test

    International Nuclear Information System (INIS)

    Karube, Koji; Akagi, Shinji; Terano, Toshihiro; Onuki, Osamu; Ito, Hideaki; Aoki, Hiroshi; Odo, Toshihiro

    2004-03-01

    This paper describes the results of primary auxiliary cooling system, which were done as a part of JOYO MK-III function test. The aim of the tests was to confirm the operational performance of primary auxiliary EMP and the protection system including siphon breaker of primary auxiliary cooling system. The items of the tests were: (Test No.): (Test item). 1) SKS-117: EMP start up test. 2) SKS-118-1: EMP start up test when pony motor running. 3) SKS-121: Function test of siphon breaker. The results of the tests satisfied the required performance, and demonstrated successful operation of primary auxiliary cooling system. (author)

  17. Effect of embryo density on in vitro development and gene expression in bovine in vitro-fertilized embryos cultured in a microwell system.

    Science.gov (United States)

    Sugimura, Satoshi; Akai, Tomonori; Hashiyada, Yutaka; Aikawa, Yoshio; Ohtake, Masaki; Matsuda, Hideo; Kobayashi, Shuji; Kobayashi, Eiji; Konishi, Kazuyuki; Imai, Kei

    2013-01-01

    To identify embryos individually during in vitro development, we previously developed the well-of-the-well (WOW) dish, which contains 25 microwells. Here we investigated the effect of embryo density (the number of embryos per volume of medium) on in vitro development and gene expression of bovine in vitro-fertilized embryos cultured in WOW dishes. Using both conventional droplet and WOW culture formats, 5, 15, and 25 bovine embryos were cultured in 125 μl medium for 168 h. The blastocysts at Day 7 were analyzed for number of cells and expression of ten genes (CDX2, IFN-tau, PLAC8, NANOG, OCT4, SOX2, AKR1B1, ATP5A1, GLUT1 and IGF2R). In droplet culture, the rates of formation of >4-cell cleavage embryos and blastocysts were significantly lower in embryos cultured at 5 embryos per droplet than in those cultured at 15 or 25 embryos per droplet, but not in WOW culture. In both droplet and WOW culture, developmental kinetics and blastocyst cell numbers did not differ among any groups. IFN-tau expression in embryos cultured at 25 embryos per droplet was significantly higher than in those cultured at 15 embryos per droplet and in artificial insemination (AI)-derived blastocysts. Moreover, IGF2R expression was significantly lower in the 25-embryo group than in the 5-embryo group and in AI-derived blastocysts. In WOW culture, these expressions were not affected by embryo density and were similar to those in AI-derived blastocysts. These results suggest that, as compared with conventional droplet culture, in vitro development and expression of IFN-tau and IGF2R in the microwell system may be insensitive to embryo density.

  18. Combined in vivo and in vitro approach for the characterization of penicillin-specific polyclonal lymphocyte reactivity: tolerance tests with safe penicillins instead of challenge with culprit drugs.

    Science.gov (United States)

    Sachs, B; Al Masaoudi, T; Merk, H F; Erdmann, S

    2004-10-01

    Amino-penicillins are a major cause of delayed-type reactions to penicillins. The aim of this study was to establish a diagnostic approach for the characterization of the individual penicillin-specific polyclonal lymphocyte reactivity in order to detect side chain-specific sensitization to amino-penicillins. Patients can then be advised to undergo a tolerance test with safe penicillins instead of provocation with culprit penicillins for confirmation of penicillin allergy. We investigated penicillin-specific polyclonal lymphocyte reactivity in nine patients with delayed-type reactions to amino-penicillins by a combined in vivo (patch, prick and intracutaneous tests with delayed readings) and in vitro (lymphocyte transformation test, LTT) approach. A combination of LTT and skin tests improved the sensitivity for the characterization of penicillin-specific polyclonal lymphocyte reactivity and allowed the detection of three different patterns of lymphocyte reactivity. Four patients showed a side chain-specific sensitization to amino-penicillins in vivo and in vitro and were advised to undergo tolerance tests with safe penicillins. Two patients agreed and were exposed to parenteral benzyl-penicillin and oral phenoxymethyl-penicillin which they tolerated without complications. These data suggest that a combined in vivo and in vitro approach is helpful for the detection of side chain-specific sensitization to amino-penicillins. Patients with such sensitization are very likely to tolerate safe penicillins, thereby expanding their therapeutic options when antibiotic treatment is required.

  19. AlgiMatrix™ based 3D cell culture system as an in-vitro tumor model for anticancer studies.

    Directory of Open Access Journals (Sweden)

    Chandraiah Godugu

    Full Text Available Three-dimensional (3D in-vitro cultures are recognized for recapitulating the physiological microenvironment and exhibiting high concordance with in-vivo conditions. Taking the advantages of 3D culture, we have developed the in-vitro tumor model for anticancer drug screening.Cancer cells grown in 6 and 96 well AlgiMatrix™ scaffolds resulted in the formation of multicellular spheroids in the size range of 100-300 µm. Spheroids were grown in two weeks in cultures without compromising the growth characteristics. Different marketed anticancer drugs were screened by incubating them for 24 h at 7, 9 and 11 days in 3D cultures and cytotoxicity was measured by AlamarBlue® assay. Effectiveness of anticancer drug treatments were measured based on spheroid number and size distribution. Evaluation of apoptotic and anti-apoptotic markers was done by immunohistochemistry and RT-PCR. The 3D results were compared with the conventional 2D monolayer cultures. Cellular uptake studies for drug (Doxorubicin and nanoparticle (NLC were done using spheroids.IC(50 values for anticancer drugs were significantly higher in AlgiMatrix™ systems compared to 2D culture models. The cleaved caspase-3 expression was significantly decreased (2.09 and 2.47 folds respectively for 5-Fluorouracil and Camptothecin in H460 spheroid cultures compared to 2D culture system. The cytotoxicity, spheroid size distribution, immunohistochemistry, RT-PCR and nanoparticle penetration data suggested that in vitro tumor models show higher resistance to anticancer drugs and supporting the fact that 3D culture is a better model for the cytotoxic evaluation of anticancer drugs in vitro.The results from our studies are useful to develop a high throughput in vitro tumor model to study the effect of various anticancer agents and various molecular pathways affected by the anticancer drugs and formulations.

  20. BMFT-UPTF densitometer system test report

    International Nuclear Information System (INIS)

    Menkhaus, D.E.

    1985-11-01

    This report documents acceptance test results performed on the five Upper Plenum Test Facility (UPTF) three-beam densitometer systems and spare parts. The five densitometer systems are used on the UPTF four hot legs and broken cold leg to measure average chordal-beam densities. The primary objectives of the tests performed were: to verify all assemblies fit as designed (mechanical fitup); to ensure radiation levels met the criteria (<2.5 mR/h); to verify that design accuracy requirements were met (performance tests); and to verify proper operation of the densitometer systems (functional checks). 15 figs., 11 tabs

  1. [Laboratory accreditation and proficiency testing].

    Science.gov (United States)

    Kuwa, Katsuhiko

    2003-05-01

    ISO/TC 212 covering clinical laboratory testing and in vitro diagnostic test systems will issue the international standard for medical laboratory quality and competence requirements, ISO 15189. This standard is based on the ISO/IEC 17025, general requirements for competence of testing and calibration laboratories and ISO 9001, quality management systems-requirements. Clinical laboratory services are essential to patient care and therefore should be available to meet the needs of all patients and clinical personnel responsible for human health care. If a laboratory seeks accreditation, it should select an accreditation body that operates according to this international standard and in a manner which takes into account the particular requirements of clinical laboratories. Proficiency testing should be available to evaluate the calibration laboratories and reference measurement laboratories in clinical medicine. Reference measurement procedures should be of precise and the analytical principle of measurement applied should ensure reliability. We should be prepared to establish a quality management system and proficiency testing in clinical laboratories.

  2. System Testing of Desktop and Web Applications

    Science.gov (United States)

    Slack, James M.

    2011-01-01

    We want our students to experience system testing of both desktop and web applications, but the cost of professional system-testing tools is far too high. We evaluate several free tools and find that AutoIt makes an ideal educational system-testing tool. We show several examples of desktop and web testing with AutoIt, starting with simple…

  3. IN VITRO MODELS TO EVALUATE DRUG-INDUCED HYPERSENSITIVITY: POTENTIAL TEST BASED ON ACTIVATION OF DENDRITIC CELLS

    Directory of Open Access Journals (Sweden)

    Valentina Galbiati

    2016-07-01

    Full Text Available Hypersensitivity drug reactions (HDRs are the adverse effect of pharmaceuticals that clinically resemble allergy. HDRs account for approximately 1/6 of drug-induced adverse effects, and include immune-mediated ('allergic' and non immune-mediated ('pseudo allergic' reactions. In recent years, the severe and unpredicted drug adverse events clearly indicate that the immune system can be a critical target of drugs. Enhanced prediction in preclinical safety evaluation is, therefore, crucial. Nowadays, there are no validated in vitro or in vivo methods to screen the sensitizing potential of drugs in the pre-clinical phase. The problem of non-predictability of immunologically-based hypersensitivity reactions is related to the lack of appropriate experimental models rather than to the lack of -understanding of the adverse phenomenon.We recently established experimental conditions and markers to correctly identify drug associated with in vivo hypersensitivity reactions using THP-1 cells and IL-8 production, CD86 and CD54 expression. The proposed in vitro method benefits from a rationalistic approach with the idea that allergenic drugs share with chemical allergens common mechanisms of cell activation. This assay can be easily incorporated into drug development for hazard identification of drugs, which may have the potential to cause in vivo hypersensitivity reactions. The purpose of this review is to assess the state of the art of in vitro models to assess the allergenic potential of drugs based on the activation of dendritic cells.

  4. Test software for BESIII MDC electronics system

    International Nuclear Information System (INIS)

    Zhang Hongyu; Sheng Huayi; Zhu Haitao; Ji Xiaolu; Zhao Dongxu

    2006-01-01

    This paper presents the design of Test System Software for BESIII MDC Electronics. Two kinds of test systems, SBS VP7 based and PowerPC based systems, and their corresponding test software are introduced. The software is developed in LabVIEW 7.1 and Microsoft Visual C++ 6.0, some test functions of the software, as well as their user interfaces, are described in detail. The software has been applied in hardware debugging, performance test and long term stability test. (authors)

  5. Preoperational test report, primary ventilation system

    Energy Technology Data Exchange (ETDEWEB)

    Clifton, F.T.

    1997-11-04

    This represents a preoperational test report for Primary Ventilation Systems, Project W-030. Project W-030 provides a ventilation upgrade for the four Aging Waste Facility tanks. The system provides vapor space filtered venting of tanks AY101, AY102, AZ101, AZ102. The tests verify correct system operation and correct indications displayed by the central Monitor and Control System.

  6. Preoperational test report, primary ventilation system

    International Nuclear Information System (INIS)

    Clifton, F.T.

    1997-01-01

    This represents a preoperational test report for Primary Ventilation Systems, Project W-030. Project W-030 provides a ventilation upgrade for the four Aging Waste Facility tanks. The system provides vapor space filtered venting of tanks AY101, AY102, AZ101, AZ102. The tests verify correct system operation and correct indications displayed by the central Monitor and Control System

  7. Characterization of DNA repair phenotypes of Xeroderma pigmentosum cell lines by a paralleled in vitro test; Phenotypage de la reparation de l'ADN de lignees Xeroderma pigmentosum, par un test in vitro multiparametrique

    Energy Technology Data Exchange (ETDEWEB)

    Raffin, A.L.

    2009-06-15

    DNA is constantly damaged modifying the genetic information for which it encodes. Several cellular mechanisms as the Base Excision Repair (BER) and the Nucleotide Excision Repair (NER) allow recovering the right DNA sequence. The Xeroderma pigmentosum is a disease characterised by a deficiency in the NER pathway. The aim of this study was to propose an efficient and fast test for the diagnosis of this disease as an alternative to the currently available UDS test. DNA repair activities of XP cell lines were quantified using in vitro miniaturized and paralleled tests in order to establish DNA repair phenotypes of XPA and XPC deficient cells. The main advantage of the tests used in this study is the simultaneous measurement of excision or excision synthesis (ES) of several lesions by only one cellular extract. We showed on one hand that the relative ES of the different lesions depend strongly on the protein concentration of the nuclear extract tested. Working at high protein concentration allowed discriminating the XP phenotype versus the control one, whereas it was impossible under a certain concentration's threshold. On the other hand, while the UVB irradiation of control cells stimulated their repair activities, this effect was not observed in XP cells. This study brings new information on the XPA and XPC protein roles during BER and NER and underlines the complexity of the regulations of DNA repair processes. (author)

  8. Successful validation of in vitro methods in toxicology by ZEBET, the National Centre for Alternatives in Germany at the BfR (Federal Institute for Risk Assessment).

    Science.gov (United States)

    Spielmann, Horst; Grune, Barbara; Liebsch, Manfred; Seiler, Andrea; Vogel, Richard

    2008-06-01

    A short description of the history of the 3Rs concept is given, which was developed as the scientific concept to refine, reduce and replace animal experiments by Russel and Burch more than 40 years ago. In addition, the legal framework in Europe for developing alternatives to animal experiments is given and the current status of in vitro systems in pharmacology and toxicology is described including an update on metabolising systems. The decrease in experimental animal numbers during the past decade in Europe is illustrated by the situation in Germany and the contribution of international harmonisation of test guidelines on reducing animal numbers in regulatory testing is described. A review of the development of the principles of experimental validation is given and the 3T3 NRU in vitro phototoxicity test is used as an example for a successful validation study, which led to the acceptance of the first in vitro toxicity test for regulatory purposes by the OECD. Finally, the currently accepted alternative methods for standardisation and safety testing of drugs, biologicals and medical devices are summarised.

  9. Operational test report, integrated system test (ventilation upgrade)

    International Nuclear Information System (INIS)

    HARTY, W.M.

    1999-01-01

    Operational Final Test Report for Integrated Systems, Project W-030 (Phase 2 test, RECIRC and HIGH-HEAT Modes). Project W-030 provides a ventilation upgrade for the four Aging Waste Facility tanks, including upgraded vapor space cooling and filtered venting of tanks AY101, AY102, AZ101, AZ102

  10. Operational test report integrated system test (ventilation upgrade)

    Energy Technology Data Exchange (ETDEWEB)

    HARTY, W.M.

    1999-10-05

    Operational Final Test Report for Integrated Systems, Project W-030 (Phase 2 test, RECIRC and HIGH-HEAT Modes). Project W-030 provides a ventilation upgrade for the four Aging Waste Facility tanks, including upgraded vapor space cooling and filtered venting of tanks AY101, Ay102, AZ101, AZ102.

  11. Ship Systems Survivability Test Site

    Data.gov (United States)

    Federal Laboratory Consortium — Area for testing survivability of shipboard systems to include electrical, communications, and fire suppression. Multipurpose test range for supporting gun firing,...

  12. The 21th quality control survey for radioisotope in vitro tests in Japan, 1999

    Energy Technology Data Exchange (ETDEWEB)

    Shishiba, Yoshimasa; Shimizu, Taeko [Toranomon Hospital, Tokyo (Japan); Ichihara, Kiyoshi; Kousaka, Tadako; Kobayashi, Hisae; Tachibana, Katsuhiko; Tsushima, Toshio; Hoshino, Minoru; Mori, Mikio

    2000-10-01

    The quality control survey in the title has been conducted every year from 1978 to improve the quality of the in vitro tests and this report gives the survey results done in 1999. The survey was performed in 133 facilities out of 283 in Japan, which involved 18 national and public university hospitals, 16 private university hospitals, 19 national and public hospitals, 18 private hospitals, 41 hygiene test institutes and 21 reagent manufacturers. Tests examined were on 6 substances related to functions of pituitary, 5 of thyroid function, 1 of parathyroid, 4 of digestive-pancreas, 5 of gonad-placenta, 4 of adrenal and 1 of renal-blood pressure regulation, on IgE, on digoxin and on 11 tumor-related substances. Tests were done on 2 - 3 samples supplied from the Committee and for reproducibility for within-day, between-day, effects of freeze-thaw and periodic variation after thawing. Methods involved those using non-radioisotopes like enzyme immunoassay as well as those using radioactive isotopes like radioimmunoassay. Results given by the mean, standard deviation and coefficient of variation for each kit, were analyzed for within-kit (between-facility) variation and between-kit (between-reagent) variation by one way analysis of variance. Findings were similar to those previously reported without the particular new problem; however, the committee considered that the survey should be done with human serum as a matrix especially for samples with a high matrix effect in future. (K.H.)

  13. In vitro developmental toxicity test detects inhibition of stem cell differentiation by silica nanoparticles

    International Nuclear Information System (INIS)

    Park, Margriet V.D.Z.; Annema, Wijtske; Salvati, Anna; Lesniak, Anna; Elsaesser, Andreas; Barnes, Clifford; McKerr, George; Howard, C. Vyvyan; Lynch, Iseult; Dawson, Kenneth A.; Piersma, Aldert H.; Jong, Wim H. de

    2009-01-01

    While research into the potential toxic properties of nanomaterials is now increasing, the area of developmental toxicity has remained relatively uninvestigated. The embryonic stem cell test is an in vitro screening assay used to investigate the embryotoxic potential of chemicals by determining their ability to inhibit differentiation of embryonic stem cells into spontaneously contracting cardiomyocytes. Four well characterized silica nanoparticles of various sizes were used to investigate whether nanomaterials are capable of inhibition of differentiation in the embryonic stem cell test. Nanoparticle size distributions and dispersion characteristics were determined before and during incubation in the stem cell culture medium by means of transmission electron microscopy (TEM) and dynamic light scattering. Mouse embryonic stem cells were exposed to silica nanoparticles at concentrations ranging from 1 to 100 μg/ml. The embryonic stem cell test detected a concentration dependent inhibition of differentiation of stem cells into contracting cardiomyocytes by two silica nanoparticles of primary size 10 (TEM 11) and 30 (TEM 34) nm while two other particles of primary size 80 (TEM 34) and 400 (TEM 248) nm had no effect up to the highest concentration tested. Inhibition of differentiation of stem cells occurred below cytotoxic concentrations, indicating a specific effect of the particles on the differentiation of the embryonic stem cells. The impaired differentiation of stem cells by such widely used particles warrants further investigation into the potential of these nanoparticles to migrate into the uterus, placenta and embryo and their possible effects on embryogenesis.

  14. Internal testing of pipe systems with IRIS inspection system

    International Nuclear Information System (INIS)

    1986-01-01

    The internal piping inspection system IRIS allows inside testing of pipes with an internal diameter of NW 70 as a minimum, and of any horizontal or vertical layout of the piping system. Visual testing is done by means of an integrated CCD video system with high resolution power. Technical data are given and examples of applications, in the German and English language. (DG) [de

  15. Test results of six-month test of two water electrolysis systems

    Science.gov (United States)

    Mills, E. S.; Wells, G. W.

    1972-01-01

    The two water electrolysis systems used in the NASA space station simulation 90-day manned test of a regenerative life support system were refurbished as required and subjected to 26-weeks of testing. The two electrolysis units are both promising systems for oxygen and hydrogen generation and both needed extensive long-term testing to evaluate the performance of the respective cell design and provide guidance for further development. Testing was conducted to evaluate performance in terms of current, pressure, variable oxygen demands, and orbital simulation. An automatic monitoring system was used to record, monitor and printout performance data at one minute, ten minute or one-hour intervals. Performance data is presented for each day of system operation for each module used during the day. Failures are analyzed, remedial action taken to eliminate problems is discussed and recommendations for redesign for future space applications are stated.

  16. System design description for the whole element furnace testing system

    International Nuclear Information System (INIS)

    Ritter, G.A.; Marschman, S.C.; MacFarlan, P.J.; King, D.A.

    1998-05-01

    This document provides a detailed description of the Hanford Spent Nuclear Fuel (SNF) Whole Element Furnace Testing System located in the Postirradiation Testing Laboratory G-Cell (327 Building). Equipment specifications, system schematics, general operating modes, maintenance and calibration requirements, and other supporting information are provided in this document. This system was developed for performing cold vacuum drying and hot vacuum drying testing of whole N-Reactor fuel elements, which were sampled from the 105-K East and K West Basins. The proposed drying processes are intended to allow dry storage of the SNF for long periods of time. The furnace testing system is used to evaluate these processes by simulating drying sequences with a single fuel element and measuring key system parameters such as internal pressures, temperatures, moisture levels, and off-gas composition

  17. Strategy for the hemocompatibility testing of microparticles.

    Science.gov (United States)

    Braune, S; Basu, S; Kratz, K; Johansson, J Bäckemo; Reinthaler, M; Lendlein, A; Jung, F

    2016-01-01

    Polymer-based microparticles are applied as non-thrombogenic or thrombogenic materials in a wide variety of intra- or extra-corporeal medical devices. As demanded by the regulatory agencies, the hemocompatibility of these blood contacting biomaterials has to be evaluated in vitro to ensure that the particle systems appropriately fulfill the envisioned function without causing undesired events such as thrombosis or inflammation. Currently described in vitro assays for hemocompatibility testing of particles comprise tests with different single cell types (e.g. erythrocytes or leukocytes), varying concentrations/dilutions of the used blood cells or whole blood, which are not standardized.Here, we report about an in vitro dynamic test system for studying the hemocompatibility of polymeric microparticles utilizing fresh human whole blood from apparently healthy subjects, collected and processed under standardized conditions. Spherical poly(ether imide) microparticles with an average diameter of 140±30 μm were utilized as model systems. Reported as candidate materials for the removal of uremic toxins, these microparticles are anticipated to facilitate optimal flow conditions in a dialyzer with minimal backflow and blood cell damage. Pristine (PEI) and potassium hydroxide (PEI-KOH) functionalized microparticles exhibited similarly nanoporous surfaces (PEI: ØExternal pore = 90±60 nm; PEI-KOH ØExternal pore = 150±130 nm) but varying water wettabilities (PEI: θadv = 112±10° PEI-KOH θadv = 60±2°). The nanoporosity of the microparticle surfaces allows the exchange of toxic solutes from blood towards the interconnective pores in the particle core, while an immigration of the substantially larger blood cells is inhibited.Sterilized PEI microparticles were incorporated -air-free -in a syringe-based test system and exposed to whole blood for 60 minutes under gentle agitation. Thereafter, thrombi formation on the particles surfaces were analyzed

  18. Degradation behaviour of LAE442-based plate–screw-systems in an in vitro bone model

    International Nuclear Information System (INIS)

    Wolters, Leonie; Besdo, Silke; Angrisani, Nina; Wriggers, Peter; Hering, Britta; Seitz, Jan-Marten; Reifenrath, Janin

    2015-01-01

    The use of absorbable implant materials for fixation after bone fracture helps to avoid a second surgery for implant removal and the risks and costs involved. Magnesium (Mg) is well known as a potential metallic material for degradable implants. The aim of the present in vitro study was to evaluate if degradable LAE442-based magnesium plate–screw-systems are suitable candidates for osteosynthesis implants in load-bearing bones. The corrosion behaviour was tested concerning the influence of different surface treatments, coatings and screw torques. Steel plates and screws of the same size served as control. Plates without special treatment screwed on up to a specified torque of 15 cNm or 7 cNm, NaOH treated plates (15 cNm), magnesium fluoride coated plates (15 cNm) and steel plates as control (15 cNm) were examined in pH-buffered, temperature-controlled SBF solution for two weeks. The experimental results indicate that the LAE442 plates and screws coated with magnesium fluoride revealed a lower hydrogen evolution in SBF solution as well as a lower weight loss and volume decrease in μ-computed tomography (μCT). The nanoindentation and SEM/EDX measurements at several plate areas showed no significant differences. Summarized, the different screw torques did not affect the corrosion behaviour differently. Also the NaOH treatment seemed to have no essential influence on the degradation kinetics. The plates coated with magnesium fluoride showed a decreased corrosion rate. Hence, it is recommended to consider this coating for the next in vivo study. - Highlights: • Mg-based plate screw systems were examined in an in vitro corrosion setup. • Different screw torques did not affect the corrosion behaviour. • Pretreatment with NaOH showed no increase in corrosion resistance. • Fluoride coating slowed down the corrosion rate of plates. • Fluoride coating might be an alternative for decrease of corrosion rate in vivo

  19. Degradation behaviour of LAE442-based plate–screw-systems in an in vitro bone model

    Energy Technology Data Exchange (ETDEWEB)

    Wolters, Leonie [Small Animal Clinic, University of Veterinary Medicine Hannover, Foundation, Bünteweg 9, 30559 Hannover (Germany); Besdo, Silke [Institute of Continuum Mechanics, Leibniz Universität Hannover, Appelstraße 11, 30167 Hannover (Germany); Angrisani, Nina [Small Animal Clinic, University of Veterinary Medicine Hannover, Foundation, Bünteweg 9, 30559 Hannover (Germany); Wriggers, Peter [Institute of Continuum Mechanics, Leibniz Universität Hannover, Appelstraße 11, 30167 Hannover (Germany); Hering, Britta [Institute of Production Engineering and Machine Tools, Leibniz Universität Hannover, An der Universität 2, 30823 Garbsen (Germany); Seitz, Jan-Marten [Institute of Materials Science, Leibniz Universität Hannover, An der Universität 2, 30823 Garbsen (Germany); Reifenrath, Janin, E-mail: janin.reifenrath@tiho-hannover.de [Small Animal Clinic, University of Veterinary Medicine Hannover, Foundation, Bünteweg 9, 30559 Hannover (Germany)

    2015-04-01

    The use of absorbable implant materials for fixation after bone fracture helps to avoid a second surgery for implant removal and the risks and costs involved. Magnesium (Mg) is well known as a potential metallic material for degradable implants. The aim of the present in vitro study was to evaluate if degradable LAE442-based magnesium plate–screw-systems are suitable candidates for osteosynthesis implants in load-bearing bones. The corrosion behaviour was tested concerning the influence of different surface treatments, coatings and screw torques. Steel plates and screws of the same size served as control. Plates without special treatment screwed on up to a specified torque of 15 cNm or 7 cNm, NaOH treated plates (15 cNm), magnesium fluoride coated plates (15 cNm) and steel plates as control (15 cNm) were examined in pH-buffered, temperature-controlled SBF solution for two weeks. The experimental results indicate that the LAE442 plates and screws coated with magnesium fluoride revealed a lower hydrogen evolution in SBF solution as well as a lower weight loss and volume decrease in μ-computed tomography (μCT). The nanoindentation and SEM/EDX measurements at several plate areas showed no significant differences. Summarized, the different screw torques did not affect the corrosion behaviour differently. Also the NaOH treatment seemed to have no essential influence on the degradation kinetics. The plates coated with magnesium fluoride showed a decreased corrosion rate. Hence, it is recommended to consider this coating for the next in vivo study. - Highlights: • Mg-based plate screw systems were examined in an in vitro corrosion setup. • Different screw torques did not affect the corrosion behaviour. • Pretreatment with NaOH showed no increase in corrosion resistance. • Fluoride coating slowed down the corrosion rate of plates. • Fluoride coating might be an alternative for decrease of corrosion rate in vivo.

  20. Excitation system testing in HPP 'Uvac'

    Directory of Open Access Journals (Sweden)

    Milojčić Nemanja

    2011-01-01

    Full Text Available The excitation system of hydro unit in HPP 'Uvac' and results of testings of excitation system performed for achieving of unit's mathematical model are presented in this paper. Description of excitation system equipment, parameters of regulators and results obtained after testings are presented. The presented results showed that the regulators are properly adjusted and that the excitation system is completely functional and reliable.

  1. Coupling auto trophic in vitro plant cultivation system to scanning electron microscope to study plant-fungal interactions

    Energy Technology Data Exchange (ETDEWEB)

    Jaeger, N. de; Decock, C.; Declereck, S.; Providencia, I. E. de la

    2010-07-01

    The interactions of plants with pathogens and beneficial micro-organisms have been seldom compared on the same host and under strict controlled auto trophic in vitro culture conditions. Here, the life cycle of two plant beneficial (Glomus sp. MUCL 41833 and Trichoderma harzianum) and one plant pathogen (Rhizoctonia solani) fungi were described on potato (Solanum tuberosum) plantlets under auto trophic in vitro culture conditions using video camera imaging and the scanning electron microscope (SEM). (i) The colony developmental pattern of the extraradical mycelium within the substrate, (ii) the reproduction structures and (iii) the three-dimensional spatial arrangements of the fungal hyphae within the potato root cells were successfully visualized, monitored and described. The combination of the autotrophic in vitro culture system and SEM represent a powerful tool for improving our knowledge on the dynamics of plant-fungal interactions. (Author) 41 refs.

  2. The Systems Test Architect: Enabling The Leap From Testable To Tested

    Science.gov (United States)

    2016-09-01

    is and it would require some “fine-tuning.” [w] Mr. Peart finally has a physical architecture, and feels confident that it will meet the desired ...test and evaluation in systems engineering, this thesis examines the concept that test and evaluation, based on its desired early involvement in the...the documented roles of test and evaluation in systems engineering, this thesis examines the concept that test and evaluation, based on its desired

  3. In vitro drug sensitivity testing of tumor cells from patients with non-Hodgkin's lymphoma using the fluorometric microculture cytotoxicity assay.

    Science.gov (United States)

    Nygren, P; Hagberg, H; Glimelius, B; Sundström, C; Kristensen, J; Christiansen, I; Larsson, R

    1994-01-01

    Tumor cell drug sensitivity is an important determinant of chemotherapy response. Its measurement in vitro would aid in therapy individualization and new drug development. The fluorometric microculture cytotoxicity assay (FMCA), based on production by viable cells of fluorescent fluorescein after 3 days of culture, was used for cytotoxic drug sensitivity testing of 73 samples of tumor cells from patients with non-Hodgkin's lymphoma (NHL). The technical success rate was 92%, and FMCA data showed good correlation to the Disc assay. NHL samples were considerably more drug sensitive than were samples from in vivo resistant tumors. There was no obvious difference in drug sensitivity for high- vs. low-grade or untreated vs. previously treated low-grade NHL. For 26 patients, clinical outcome was correlated to in vitro response giving a sensitivity and specificity of 93 and 48%, respectively. Cross-resistance between standard drugs was frequent in vitro. Resistance modulators potentiated the effect of vincristine and doxorubicin in 10-29% of the samples, most frequently from previously treated patients. The FMCA seems to report clinically relevant drug sensitivity data for NHL, and thus it could serve as a tool for optimization of chemotherapy in the future.

  4. 21 CFR 862.3660 - Phenobarbital test system.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Phenobarbital test system. 862.3660 Section 862....3660 Phenobarbital test system. (a) Identification. A phenobarbitol test system is a device intended to measure phenobarbital, an antiepileptic and sedative-hypnotic drug, in human specimens. Measurements...

  5. Assessment of in vitro radiosensitivity of human peripheral blood lymphocytes

    International Nuclear Information System (INIS)

    Knox, S.J.; Shifrine, M.; Rosenblatt, L.S.

    1980-01-01

    The proliferative capacity of sensitive lymphocyte progenitor cells, from thirty-one clinically normal adults, was evaluated following in vitro x-irradiation (0-400R). Radiation effects were studied using both whole blood and lymphocyte-enriched mononuclear cell fractions in the lymphocyte stimulation test and colony formation assay with 6 different mitogens and antigens. Radiation dose-response survival curves were determined for the different test groups. The sensitivity of the different assay systems is compared and normative values are presented that may be used for comparison purposes to determine the relative radiosensitivity of atypical individuals and groups of individuals

  6. Acceptance test procedure, 241-SY-101 Flexible Receiver System, Phase III testing

    International Nuclear Information System (INIS)

    Ritter, G.A.

    1994-01-01

    This Acceptance Test Procedure is for the 241-SY-101 Flexible Receiver System, Phase III Testing. This procedure will test the sealing integrity of the Flexible Receiver System to ensure that release of waste and aerosols will be minimized during the removal of the test mixer pump from tank SY-101

  7. Programmable, automated transistor test system

    Science.gov (United States)

    Truong, L. V.; Sundburg, G. R.

    1986-01-01

    A programmable, automated transistor test system was built to supply experimental data on new and advanced power semiconductors. The data will be used for analytical models and by engineers in designing space and aircraft electric power systems. A pulsed power technique was used at low duty cycles in a nondestructive test to examine the dynamic switching characteristic curves of power transistors in the 500 to 1000 V, 10 to 100 A range. Data collection, manipulation, storage, and output are operator interactive but are guided and controlled by the system software.

  8. Considering the antibacterial activity of Zataria multiflora Boiss essential oil treated with gamma-irradiation in vitro and in vivo systems

    International Nuclear Information System (INIS)

    Fatemi Faezeh; Dini Salome; Dadkhah Abolfazl; Zolfaghari Mohammad Reza

    2015-01-01

    The aim of the present study was to evaluate the antibacterial activities of essential oils (EOs) obtained from the aerial parts of Zataria multiflora Boiss against Bacillus cereus, Pseudomonas aeroginosa, Escherichia coli and Staphylococcus aureus by in vivo and in vitro methods. Also, the effects of gamma-irradiation (0, 10 and 25 kGy) as a new microbial decontamination on the antibacterial activities of Z. multiflora were examined. For this purpose, the collected herbs were exposed to radiation at doses of 0, 10 and 25 kGy following essential oil (EOs) extraction by steam distillation. Then, the in vitro antibacterial potency of the irradiated and non-irradiated oils was determined by using disc diffusion, agar well diffusion and MIC and MBC determination assays. The in vivo antibacterial activity was also studied in sepsis model induced by CLP surgery by Colony forming units (CFUs) determination. The results showed that the extracted oils were discovered to be effective against all the gram positive and gram negative pathogens in vitro system. In addition, the oil significantly diminished the increased CFU count observed in CLP group. Moreover, the irradiated samples were found to possess the antibacterial activities as the non-irradiated ones both in vitro and in vivo systems. These data indicated the potential use of gamma-irradiation as a safe technique for preservation of Z. multiflora as a medicinal plant with effective antibacterial activities. - Highlights: • Zataria multiflora Boiss essential oil has potential in vitro antimicrobial effect. • Z. multiflora oil has potential antimicrobial effect in vivo system. • The antibacterial activities of the oil remained after irradiation treatments

  9. Comparative performance analysis of human iPSC-derived and primary neural progenitor cells (NPC grown as neurospheres in vitro

    Directory of Open Access Journals (Sweden)

    Maxi Hofrichter

    2017-12-01

    hiPSC-NPCs-derived neurospheres seem to be useful for DNT evaluation representing early neural development in vitro. More system characterization by compound testing is needed to gain higher confidence in this method.

  10. Filling the concept with data: integrating data from different in vitro and in silico assays on skin sensitizers to explore the battery approach for animal-free skin sensitization testing.

    Science.gov (United States)

    Natsch, Andreas; Emter, Roger; Ellis, Graham

    2009-01-01

    Tests for skin sensitization are required prior to the market launch of new cosmetic ingredients. Significant efforts are made to replace the current animal tests. It is widely recognized that this cannot be accomplished with a single in vitro test, but that rather the integration of results from different in vitro and in silico assays will be needed for the prediction of the skin sensitization potential of chemicals. This has been proposed as a theoretical scheme so far, but no attempts have been made to use experimental data to prove the validity of this concept. Here we thus try for the first time to fill this widely cited concept with data. To this aim, we integrate and report both novel and literature data on 116 chemicals of known skin sensitization potential on the following parameters: (1) peptide reactivity as a surrogate for protein binding, (2) induction of antioxidant/electrophile responsive element dependent luciferase activity as a cell-based assay; (3) Tissue Metabolism Simulator skin sensitization model in silico prediction; and (4) calculated octanol-water partition coefficient. The results of the in vitro assays were scaled into five classes from 0 to 4 to give an in vitro score and compared to the local lymph node assay (LLNA) data, which were also scaled from 0 to 4 (nonsensitizer/weak/moderate/strong/extreme). Different ways of evaluating these data have been assessed to rate the hazard of chemicals (Cooper statistics) and to also scale their potency. With the optimized model an overall accuracy for predicting sensitizers of 87.9% was obtained. There is a linear correlation between the LLNA score and the in vitro score. However, the correlation needs further improvement as there is still a relatively high variation in the in vitro score between chemicals belonging to the same sensitization potency class.

  11. The relationship between CD86/CD54 expression and THP-1 cell viability in an in vitro skin sensitization test--human cell line activation test (h-CLAT).

    Science.gov (United States)

    Sakaguchi, Hitoshi; Ashikaga, Takao; Miyazawa, Masaaki; Kosaka, Nanae; Ito, Yuichi; Yoneyama, Katsurako; Sono, Sakiko; Itagaki, Hiroshi; Toyoda, Hidekazu; Suzuki, Hiroyuki

    2009-04-01

    Recent regulations for cosmetics in Europe prohibit animal testing for evaluating the sensitization potential of chemicals to improve animal welfare. Yet, there is not an acceptable Organization for Economic Co-operation and Development non-animal skin sensitization test method. Several in vitro skin sensitization methods that focus on the activation of Langerhans cells, including human cell lines, are being evaluated as possible alternatives. In our previous study, we optimized our human cell line activation test (h-CLAT) using THP-1 cells (monocytic leukemia cell line) and conducted an inter-laboratory study. We found that measuring CD86/CD54 expression may be useful for predicting skin sensitization. The aim of this study was to confirm the relationship between CD86/CD54 expression and THP-1 cell viability in the h-CLAT. In this study, 21 allergens (e.g., dinitrochlorobenzene, p-phenylenediamine, Ni) and 8 non-allergens (e.g., SLS, lactic acid) were evaluated. For each chemical, more than 10 concentrations that gave a predicted cell viability range of 20-95% were used. The data showed that expression patterns of CD86/CD54 differed depending on chemical. For most allergens, cytotoxicity (65-90% cell viability) was needed for enhancement of CD86/CD54 expression. The criteria of "CD86 > or = 150 or CD54 > or = 200" resulted in an accuracy of 93%, which confirms appropriate cut-off criteria for h-CLAT. Furthermore, a good correlation was observed between EC3 of local lymph node assay and EC150(CD86) or EC200(CD54) of h-CLAT (12 or 16 chemicals, respectively), which would provide a useful estimate of allergic potency. These findings suggest that h-CLAT would be a good robust in vitro skin sensitization test.

  12. Test-driven modeling of embedded systems

    DEFF Research Database (Denmark)

    Munck, Allan; Madsen, Jan

    2015-01-01

    To benefit maximally from model-based systems engineering (MBSE) trustworthy high quality models are required. From the software disciplines it is known that test-driven development (TDD) can significantly increase the quality of the products. Using a test-driven approach with MBSE may have...... a similar positive effect on the quality of the system models and the resulting products and may therefore be desirable. To define a test-driven model-based systems engineering (TD-MBSE) approach, we must define this approach for numerous sub disciplines such as modeling of requirements, use cases...... suggest that our method provides a sound foundation for rapid development of high quality system models....

  13. Rocuronium is more hepatotoxic than succinylcholine in vitro.

    Science.gov (United States)

    Sauer, Martin; Piel, Ines; Haubner, Cristof; Richter, Georg; Mann, Miriam; Nöldge-Schomburg, Gabriele; Mencke, Thomas

    2017-09-01

    The development of liver failure is a major problem in critically ill patients. The hepatotoxicity of many drugs, as one important reason for liver failure, is poorly screened for in human models. Rocuronium and succinylcholine are neuromuscular blocking agents used for tracheal intubation and for rapid-sequence induction. We used an in-vitro test with a permanent cell line and compared rocuronium and succinylcholine for hepatotoxicity. In-vitro study. A basic science laboratory, University Hospital Rostock, Germany. The basic test compound is the permanent human liver cell line HepG2/C3A. In a standardised microtitre plate assay the toxicity of different concentrations of rocuronium, succinylcholine and plasma control was tested. After two incubation periods of 3 days, the viability of cells (XTT test, lactate dehydrogenase release and trypan blue staining), micro-albumin synthesis and the cytochrome 1A2 activity (metabolism of ethoxyresorufin) were measured. Differences between rocuronium and succinylcholine were assessed using the Kruskal-Wallis one-way test and two-tailed Mann-Whitney U test. Rocuronium, but not succinylcholine, led to a significant dose-dependent decrease of viability, albumin synthesis and cytochrome 1A2 activity of test cells. An in-vitro test with a cell line showed hepatotoxicity of rocuronium that was dose-dependent. Further studies are needed to investigate the underlying mechanisms of the effects of rocuronium on hepatic cellular integrity. Not suitable.

  14. Programmable automated transistor test system

    International Nuclear Information System (INIS)

    Truong, L.V.; Sundberg, G.R.

    1986-01-01

    The paper describes a programmable automated transistor test system (PATTS) and its utilization to evaluate bipolar transistors and Darlingtons, and such MOSFET and special types as can be accommodated with the PATTS base-drive. An application of a pulsed power technique at low duty cycles in a non-destructive test is used to examine the dynamic switching characteristic curves of power transistors. Data collection, manipulation, storage, and output are operator interactive but are guided and controlled by the system software. In addition a library of test data is established on disks, tapes, and hard copies for future reference

  15. Availability of periodically tested systems

    International Nuclear Information System (INIS)

    Signoret, J.P.

    1979-01-01

    There is at the present time a need in accurate models to asess the availability of periodically tested stand-by systems. This paper shows how to improve the well known 'saw-tooth curve' model in order to take into account various reliability parameters. A model is developed to assess the pointwise and the mean availabilities of periodically tested stand-by systems. Exact and approxination formulae are given. In addition, the model developed herein leads to optimize the test interval in order to minimize the mean unavailability. A safety diesel in a nuclear power plant is given as an example

  16. The utility of in vitro solubility testing in assessment of uranium exposure

    International Nuclear Information System (INIS)

    Eidson, A.F.; Damon, E.G.; Hahn, F.F.; Griffith, W.C. Jr.

    1989-01-01

    Assessment of accidental exposures in the uranium industry requires estimation of retention and excretion rates using bioassay measurements. This task is difficult if the solubility of the inhaled uranium compound is unknown. In our research, Beagle dogs were exposed to aerosols of commercial uranium milling products containing either pure ammonium diuranate (ADU) or U 3 0 8 . Dogs were exposed to ADU aerosols to achieve a median retained body burden of 0.058 mg U per kg body weight, or to U 3 0 8 aerosols to achieve a median retained body burden of 0.28 mg U per kg. A biokinetic model was applied to simulate retention and excretion of the two forms of uranium in vivo. Comparison of published in vitro dissolution data and modelling results with information from accidental human exposures showed that in vitro dissolution studies are necessary to characterise the differential solubilities of uranium compounds, and indicate the potential for kidney toxicity. Because variability in pulmonary clearance and urinary excretion rates is high among individual people, in vitro dissolution results are only marginally useful for estimating urinary excretion rates. (author)

  17. 21 CFR 201.119 - In vitro diagnostic products.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 4 2010-04-01 2010-04-01 false In vitro diagnostic products. 201.119 Section 201...) DRUGS: GENERAL LABELING Exemptions From Adequate Directions for Use § 201.119 In vitro diagnostic products. (a) “In vitro diagnostic products” are those reagents, instruments and systems intended for use...

  18. 21 CFR 862.1225 - Creatinine test system.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Creatinine test system. 862.1225 Section 862.1225....1225 Creatinine test system. (a) Identification. A creatinine test system is a device intended to measure creatinine levels in plasma and urine. Creatinine measurements are used in the diagnosis and...

  19. 21 CFR 862.3040 - Alcohol test system.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Alcohol test system. 862.3040 Section 862.3040 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED....3040 Alcohol test system. (a) Identification. An alcohol test system is a device intented to measure...

  20. 21 CFR 862.1035 - Albumin test system.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Albumin test system. 862.1035 Section 862.1035....1035 Albumin test system. (a) Identification. An albumin test system is a device intended to measure the albumin concentration in serum and plasma. Albumin measurements are used in the diagnosis and...

  1. A tan in a test tube - in vitro models for investigating ultraviolet radiation-induced damage in skin.

    Science.gov (United States)

    Fernandez, Tara L; Dawson, Rebecca A; Van Lonkhuyzen, Derek R; Kimlin, Michael G; Upton, Zee

    2012-06-01

    Presently, global rates of skin cancers induced by ultraviolet radiation (UVR) exposure are on the rise. In view of this, current knowledge gaps in the biology of photocarcinogenesis and skin cancer progression urgently need to be addressed. One factor that has limited skin cancer research has been the need for a reproducible and physiologically-relevant model able to represent the complexity of human skin. This review outlines the main currently-used in vitro models of UVR-induced skin damage. This includes the use of conventional two-dimensional cell culture techniques and the major animal models that have been employed in photobiology and photocarcinogenesis research. Additionally, the progression towards the use of cultured skin explants and tissue-engineered skin constructs, and their utility as models of native skin's responses to UVR are described. The inherent advantages and disadvantages of these in vitro systems are also discussed. © 2012 John Wiley & Sons A/S.

  2. Open Architecture Data System for NASA Langley Combined Loads Test System

    Science.gov (United States)

    Lightfoot, Michael C.; Ambur, Damodar R.

    1998-01-01

    The Combined Loads Test System (COLTS) is a new structures test complex that is being developed at NASA Langley Research Center (LaRC) to test large curved panels and cylindrical shell structures. These structural components are representative of aircraft fuselage sections of subsonic and supersonic transport aircraft and cryogenic tank structures of reusable launch vehicles. Test structures are subjected to combined loading conditions that simulate realistic flight load conditions. The facility consists of two pressure-box test machines and one combined loads test machine. Each test machine possesses a unique set of requirements or research data acquisition and real-time data display. Given the complex nature of the mechanical and thermal loads to be applied to the various research test articles, each data system has been designed with connectivity attributes that support both data acquisition and data management functions. This paper addresses the research driven data acquisition requirements for each test machine and demonstrates how an open architecture data system design not only meets those needs but provides robust data sharing between data systems including the various control systems which apply spectra of mechanical and thermal loading profiles.

  3. Genotoxic and Antigenotoxic Assessment of Chios Mastic Oil by the In Vitro Micronucleus Test on Human Lymphocytes and the In Vivo Wing Somatic Test on Drosophila.

    Directory of Open Access Journals (Sweden)

    Dimitris Vlastos

    Full Text Available Chios mastic oil (CMO, the essential oil derived from Pistacia lentiscus (L. var. chia (Duham, has generated considerable interest because of its antimicrobial, anticancer, antioxidant and other beneficial properties. In the present study, the potential genotoxic activity of CMO as well as its antigenotoxic properties against the mutagenic agent mitomycin-C (MMC were evaluated by employing the in vitro Cytokinesis Block MicroNucleus (CBMN assay and the in vivo Somatic Mutation And Recombination Test (SMART. In the in vitro experiments, lymphocytes were treated with 0.01, 0.05 and 0.10% (v/v of CMO with or without 0.05 μg/ml MMC, while in the in vivo assay Drosophila larvae were fed with 0.05, 0.10, 0.50 and 1.00% (v/v of CMO with or without 2.50 μg/ml MMC. CMO did not significantly increase the frequency of micronuclei (MN or total wing spots, indicating lack of mutagenic or recombinogenic activity. However, the in vitro analysis suggested cytotoxic activity of CMO. The simultaneous administration of MMC with CMO did not alter considerably the frequencies of MMC-induced MN and wing spots showing that CMO doesn't exert antigenotoxic or antirecombinogenic action. Therefore, CMO could be considered as a safe product in terms of genotoxic potential. Even though it could not afford any protection against DNA damage, at least under our experimental conditions, its cytotoxic potential could be of interest.

  4. Radical scavenging activities of Rio Red grapefruits and Sour orange fruit extracts in different in vitro model systems.

    Science.gov (United States)

    Jayaprakasha, G K; Girennavar, Basavaraj; Patil, Bhimanagouda S

    2008-07-01

    Antioxidant fractions from two different citrus species such as Rio Red (Citrus paradise Macf.) and Sour orange (Citrus aurantium L.) were extracted with five different polar solvents using Soxhlet type extractor. The total phenolic content of the extracts was determined by Folin-Ciocalteu method. Ethyl acetate extract of Rio Red and Sour orange was found to contain maximum phenolics. The dried fractions were screened for their antioxidant activity potential using in vitro model systems such as 1,1-diphenyl-2-picryl hydrazyl (DPPH), phosphomolybdenum method and nitroblue tetrazolium (NBT) reduction at different concentrations. The methanol:water (80:20) fraction of Rio Red showed the highest radical scavenging activity 42.5%, 77.8% and 92.1% at 250, 500 and 1000 ppm, respectively, while methanol:water (80:20) fraction of Sour orange showed the lowest radical scavenging activity at all the tested concentrations. All citrus fractions showed good antioxidant capacity by the formation of phosphomolybdenum complex at 200 ppm. In addition, superoxide radical scavenging activity was assayed using non-enzymatic (NADH/phenaxine methosulfate) superoxide generating system. All the extracts showed variable superoxide radical scavenging activity. Moreover, methanol:water (80:20) extract of Rio Red and methanol extract of Sour orange exhibited marked reducing power in potassium ferricyanide reduction method. The data obtained using above in vitro models clearly establish the antioxidant potential of citrus fruit extracts. However, comprehensive studies need to be conducted to ascertain the in vivo bioavailability, safety and efficacy of such extracts in experimental animals. To the best of our knowledge, this is the first report on antioxidant activity of different polar extracts from Rio Red and Sour oranges.

  5. In vivo testing of chemicals that undergo substantial biotransformation: An opportunity to advance in vitro-in vivo metabolism extrapolation procedures for fish

    Science.gov (United States)

    This is one of several keynote presentations given at the beginning of a one-day workshop on in vivo testing with fish. The purpose of this talk is to describe recent progress on using in vitro data to predict metabolism impacts on chemical accumulation by fish. The focus of th...

  6. Non-Animal Testing Approach to EPA Labeling for Eye Irritation

    Science.gov (United States)

    This document is an update to EPA’s 2013 published alternative testing approach (using in vitro/ex vivo assays) for determination of eye irritation potential in the pesticide program under EPA's classification and labeling system.

  7. 21 CFR 862.1210 - Creatine test system.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Creatine test system. 862.1210 Section 862.1210....1210 Creatine test system. (a) Identification. A creatine test system is a device intended to measure creatine (a substance synthesized in the liver and pancreas and found in biological fluids) in plasma...

  8. 21 CFR 862.1065 - Ammonia test system.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Ammonia test system. 862.1065 Section 862.1065....1065 Ammonia test system. (a) Identification. An ammonia test system is a device intended to measure ammonia levels in blood, serum, and plasma, Ammonia measurements are used in the diagnosis and treatment...

  9. 21 CFR 862.3600 - Mercury test system.

    Science.gov (United States)

    2010-04-01

    ....3600 Mercury test system. (a) Identification. A mercury test system is a device intended to measure mercury, a heavy metal, in human specimens. Measurements obtained by this device are used in the diagnosis... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Mercury test system. 862.3600 Section 862.3600...

  10. Space Launch System Scale Model Acoustic Test Ignition Overpressure Testing

    Science.gov (United States)

    Nance, Donald; Liever, Peter; Nielsen, Tanner

    2015-01-01

    The overpressure phenomenon is a transient fluid dynamic event occurring during rocket propulsion system ignition. This phenomenon results from fluid compression of the accelerating plume gas, subsequent rarefaction, and subsequent propagation from the exhaust trench and duct holes. The high-amplitude unsteady fluid-dynamic perturbations can adversely affect the vehicle and surrounding structure. Commonly known as ignition overpressure (IOP), this is an important design-to environment for the Space Launch System (SLS) that NASA is currently developing. Subscale testing is useful in validating and verifying the IOP environment. This was one of the objectives of the Scale Model Acoustic Test, conducted at Marshall Space Flight Center. The test data quantifies the effectiveness of the SLS IOP suppression system and improves the analytical models used to predict the SLS IOP environments. The reduction and analysis of the data gathered during the SMAT IOP test series requires identification and characterization of multiple dynamic events and scaling of the event waveforms to provide the most accurate comparisons to determine the effectiveness of the IOP suppression systems. The identification and characterization of the overpressure events, the waveform scaling, the computation of the IOP suppression system knockdown factors, and preliminary comparisons to the analytical models are discussed.

  11. Space Launch System Scale Model Acoustic Test Ignition Overpressure Testing

    Science.gov (United States)

    Nance, Donald K.; Liever, Peter A.

    2015-01-01

    The overpressure phenomenon is a transient fluid dynamic event occurring during rocket propulsion system ignition. This phenomenon results from fluid compression of the accelerating plume gas, subsequent rarefaction, and subsequent propagation from the exhaust trench and duct holes. The high-amplitude unsteady fluid-dynamic perturbations can adversely affect the vehicle and surrounding structure. Commonly known as ignition overpressure (IOP), this is an important design-to environment for the Space Launch System (SLS) that NASA is currently developing. Subscale testing is useful in validating and verifying the IOP environment. This was one of the objectives of the Scale Model Acoustic Test (SMAT), conducted at Marshall Space Flight Center (MSFC). The test data quantifies the effectiveness of the SLS IOP suppression system and improves the analytical models used to predict the SLS IOP environments. The reduction and analysis of the data gathered during the SMAT IOP test series requires identification and characterization of multiple dynamic events and scaling of the event waveforms to provide the most accurate comparisons to determine the effectiveness of the IOP suppression systems. The identification and characterization of the overpressure events, the waveform scaling, the computation of the IOP suppression system knockdown factors, and preliminary comparisons to the analytical models are discussed.

  12. Nitrogen metabolism in a grapevine in vitro system

    Directory of Open Access Journals (Sweden)

    Nuria Llorens

    2002-09-01

    Full Text Available Ammonium, nitrate, nitrite, protein and individual and total amino acid contents were determined in grapevine (cv Sauvignon cultured in vitro. The enzyme activities of nitrate and nitrite reductases, glutamine synthetase, glutamate synthetase and dehydrogenase were also determined. The nitrogen taken up by the plants was 70% of the total nitrogen in the medium after 75 days of in vitro culture. Most of the nitrogen taken up was recovered in the leaves, yet only ammonia and amino acid concentrations were significantly higher in leaves. In roots, glutamine was the most abundant amino acid. In leaves, the most abundant amino acids were aspartate, glutamate, glutamine, alanine, arginine and g-aminobutirate. All enzyme activities were higher in roots than in leaves. These results suggest that both roots and leaves incorporate inorganic nitrogen into organic forms.

  13. 21 CFR 866.5040 - Albumin immunological test system.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Albumin immunological test system. 866.5040... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Immunological Test Systems § 866.5040 Albumin immunological test system. (a) Identification. An albumin immunological test system is a device that consists of...

  14. Development of an in vitro three dimensional loading-measurement system for long bone fixation under multiple loading conditions: a technical description

    Directory of Open Access Journals (Sweden)

    Wilson David A

    2007-11-01

    Full Text Available Abstract The purpose of this investigation was to design and verify the capabilities of an in vitro loading-measurement system that mimics in vivo unconstrained three dimensional (3D relative motion between long bone ends, applies uniform load components over the entire length of a test specimen, and measures 3D relative motion between test segment ends to directly determine test segment construct stiffness free of errors due to potting-fixture-test machine finite stiffness. Intact equine cadaveric radius bones, which were subsequently osteotomized/ostectomized and instrumented with bone plates were subjected to non-destructive axial, torsion, and 4-point bending loads through fixtures designed to allow unconstrained components of non-load associated 3D relative motion between radius ends. 3D relative motion between ends of a 50 mm long test segment was measured by an infrared optical tracking system to directly determine its stiffness. Each specimen was then loaded to ultimate failure in either torsion or bending. Cortical bone cross-section diameters and published bone biomechanical properties were substituted into classical mechanics equations to predict the intact test segment theoretical stiffness for comparison and thus loading-measurement system verification. Intact measured stiffness values were the same order of magnitude as theoretically predicted. The primary component of relative motion between ends of the test segment corresponded to that of the applied load with the other 3D components being evident and consistent in relative magnitude and direction for unconstrained loading of an unsymmetrical double plate oblique fracture configuration. Bone failure configurations were reproducible and consistent with theoretically predicted. The 3D loading-measurement system designed: a mimics unconstrained relative 3D motion between radius ends that occurs in clinical situations, b applies uniform compression, torsion, and 4-point bending loads

  15. Frequency modulation system test procedure shuttle task 501 approach and landing test configuration

    Science.gov (United States)

    Doland, G. D.

    1976-01-01

    Shuttle Task 501 is an in-line task to test the performance and compatibility of radiofrequency links between the SSO and ground, and relay via a satellite. Under Shuttle Task 501 approach and landing test (ALT) phase only a limited portion of the communication and tracking (C&T) equipment is to be tested. The principal item to be tested is a frequency modulated (FM) data link. To test this RF link, an ALT FM System was designed, constructed, and the console wiring verified. A step-by-step procedure to be used to perform the ALT FM system is presented. The ALT FM system test is to be performed prior to delivery of the equipment to the Electronic Systems Test Laboratory (ESTL).

  16. 21 CFR 862.3110 - Antimony test system.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Antimony test system. 862.3110 Section 862.3110 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES CLINICAL CHEMISTRY AND CLINICAL TOXICOLOGY DEVICES Clinical Toxicology Test Systems § 862.3110 Antimony test system. (a) Identification. A...

  17. Biofilms of vaginal Lactobacillus in vitro test.

    Science.gov (United States)

    Wei, Xiao-Yu; Zhang, Rui; Xiao, Bing-Bing; Liao, Qin-Ping

    2017-01-01

    This paper focuses on biofilms of Lactobacillus spp. - a type of normal flora isolated from healthy human vaginas of women of childbearing age; thereupon, it broadens the research scope of investigation of vaginal normal flora. The static slide culture method was adopted to foster biofilms, marked by specific fluorescence staining. Laser scanning confocal and scanning electron microscopy were used to observe the microstructure of the biofilms. Photographs taken from the microstructure were analysed to calculate the density of the biofilms. The body of Lactobacillus spp., though red, turned yellow when interacting with the green extracellular polysaccharides. The structure of the biofilm and aquaporin within the biofilm were imaged. Lactobacillus density increases over time. This study provides convincing evidence that Lactobacillus can form biofilms and grow over time in vitro. This finding establishes an important and necessary condition for selecting proper strains for the pharmaceutics of vaginal ecology.

  18. A Physical Protection Systems Test Bed for International Counter-Trafficking System Development

    International Nuclear Information System (INIS)

    Stinson, Brad J.; Kuhn, Michael J.; Donaldson, Terrence L.; Richardson, Dave; Rowe, Nathan C.; Younkin, James R.; Pickett, Chris A.

    2011-01-01

    Physical protection systems have a widespread impact on the nuclear industry in areas such as nuclear safeguards, arms control, and trafficking of illicit goods (e.g., nuclear materials) across international borders around the world. Many challenges must be overcome in design and deployment of foreign border security systems such as lack of infrastructure, extreme environmental conditions, limited knowledge of terrain, insider threats, and occasional cultural resistance. Successful security systems, whether it be a system designed to secure a single facility or a whole border security system, rely on the entire integrated system composed of multiple subsystems. This test bed is composed of many unique sensors and subsystems, including wireless unattended ground sensors, a buried fiber-optic acoustic sensor, a lossy coaxial distributed sensor, wireless links, pan-tilt-zoom cameras, mobile power generation systems, unmanned aerial vehicles, and fiber-optic-fence intrusion detection systems. A Common Operating Picture software architecture is utilized to integrate a number of these subsystems. We are currently performance testing each system for border security and perimeter security applications by examining metrics such as probability of sense and a qualitative understanding of the sensors vulnerability of defeat. The testing process includes different soil conditions for buried sensors (e.g., dry, wet, and frozen) and an array of different tests including walking, running, stealth detection, and vehicle detection. Also, long term sustainability of systems is tested including performance differences due to seasonal variations (e.g. summer versus winter, while raining, in foggy conditions). The capabilities of the test bed are discussed. Performance testing results, both at the individual component level and integrated into a larger system for a specific deployment (in situ), help illustrate the usefulness and need for integrated testing facilities to carry out this

  19. Cascade Distiller System Performance Testing Interim Results

    Science.gov (United States)

    Callahan, Michael R.; Pensinger, Stuart; Sargusingh, Miriam J.

    2014-01-01

    The Cascade Distillation System (CDS) is a rotary distillation system with potential for greater reliability and lower energy costs than existing distillation systems. Based upon the results of the 2009 distillation comparison test (DCT) and recommendations of the expert panel, the Advanced Exploration Systems (AES) Water Recovery Project (WRP) project advanced the technology by increasing reliability of the system through redesign of bearing assemblies and improved rotor dynamics. In addition, the project improved the CDS power efficiency by optimizing the thermoelectric heat pump (TeHP) and heat exchanger design. Testing at the NASA-JSC Advanced Exploration System Water Laboratory (AES Water Lab) using a prototype Cascade Distillation Subsystem (CDS) wastewater processor (Honeywell d International, Torrance, Calif.) with test support equipment and control system developed by Johnson Space Center was performed to evaluate performance of the system with the upgrades as compared to previous system performance. The system was challenged with Solution 1 from the NASA Exploration Life Support (ELS) distillation comparison testing performed in 2009. Solution 1 consisted of a mixed stream containing human-generated urine and humidity condensate. A secondary objective of this testing is to evaluate the performance of the CDS as compared to the state of the art Distillation Assembly (DA) used in the ISS Urine Processor Assembly (UPA). This was done by challenging the system with ISS analog waste streams. This paper details the results of the AES WRP CDS performance testing.

  20. Unit Testing for Command and Control Systems

    Science.gov (United States)

    Alexander, Joshua

    2018-01-01

    Unit tests were created to evaluate the functionality of a Data Generation and Publication tool for a command and control system. These unit tests are developed to constantly evaluate the tool and ensure it functions properly as the command and control system grows in size and scope. Unit tests are a crucial part of testing any software project and are especially instrumental in the development of a command and control system. They save resources, time and costs associated with testing, and catch issues before they become increasingly difficult and costly. The unit tests produced for the Data Generation and Publication tool to be used in a command and control system assure the users and stakeholders of its functionality and offer assurances which are vital in the launching of spacecraft safely.

  1. Effect of the trehalose levels on the screening of yeast as probiotic by in vivo and in vitro assays Efeito da trealose na seleção de leveduras para uso como probióticos utilizando testes in vitro e in vivo

    Directory of Open Access Journals (Sweden)

    Flaviano S. Martins

    2008-03-01

    Full Text Available Probiotics are viable defined microorganisms (bacteria or yeasts that exert a beneficial effect on the health of the host when ingested in adequate amounts. Screening for such biotherapeutic agents is commonly performed by in vitro assays simulating gastrointestinal environment to determine the ability to survive in the digestive tract. In the present study, the possibility of extrapolation of data obtained in in vitro assays to in vivo conditions was studied using five Saccharomyces cerevisiae strains isolated from Brazilian Atlantic rain forest. Trehalose contents and survival after exposure to a combination of physiological stresses generally found in the gastrointestinal tract of humans were determined for the five yeasts and compared to the behavior of Saccharomyces boulardii, a well-known probiotic. The results were completed with the colonization capacity of the gastrointestinal tract of gnotobiotic mice by these yeast strains. Some results obtained by in vitro assays are not confirmed by in vivo experiments, indicating that the extrapolation cannot be always done.Probióticos são definidos como microrganismos (bactérias e leveduras que exercem um efeito benéfico na saúde do hospedeiro quando ingeridos em quantidades adequadas. A seleção desses agentes bioterapêuticos normalmente é feita por testes in vitro simulando o ambiente gastrointestinal que determina a capacidade de sobrevivência no trato digestivo. Neste trabalho, a possibilidade de extrapolação dos dados obtidos nos testes in vitro para as condições in vivo foi estudada utilizando cinco linhagens de Saccharomyces cerevisiae isoladas da floresta Atlântica brasileira. O conteúdo de trealose e a sobrevivência após a exposição a diversos estresses fisiológicos geralmente encontrados no trato gastrointestinal de humanos foram determinados para as cinco linhagens e os resultados comparados com a Saccharomyces boulardii, um probiótico conhecido. Esses resultados

  2. KNOX1 is expressed and epigenetically regulated during in vitro conditions in Agave spp

    Directory of Open Access Journals (Sweden)

    De-la-Peña Clelia

    2012-11-01

    Full Text Available Abstract Background The micropropagation is a powerful tool to scale up plants of economical and agronomical importance, enhancing crop productivity. However, a small but growing body of evidence suggests that epigenetic mechanisms, such as DNA methylation and histone modifications, can be affected under the in vitro conditions characteristic of micropropagation. Here, we tested whether the adaptation to different in vitro systems (Magenta boxes and Bioreactors modified epigenetically different clones of Agave fourcroydes and A. angustifolia. Furthermore, we assessed whether these epigenetic changes affect the regulatory expression of KNOTTED1-like HOMEOBOX (KNOX transcription factors. Results To gain a better understanding of epigenetic changes during in vitro and ex vitro conditions in Agave fourcroydes and A. angustifolia, we analyzed global DNA methylation, as well as different histone modification marks, in two different systems: semisolid in Magenta boxes (M and temporary immersion in modular Bioreactors (B. No significant difference was found in DNA methylation in A. fourcroydes grown in either M or B. However, when A. fourcroydes was compared with A. angustifolia, there was a two-fold difference in DNA methylation between the species, independent of the in vitro system used. Furthermore, we detected an absence or a low amount of the repressive mark H3K9me2 in ex vitro conditions in plants that were cultured earlier either in M or B. Moreover, the expression of AtqKNOX1 and AtqKNOX2, on A. fourcroydes and A. angustifolia clones, is affected during in vitro conditions. Therefore, we used Chromatin ImmunoPrecipitation (ChIP to know whether these genes were epigenetically regulated. In the case of AtqKNOX1, the H3K4me3 and H3K9me2 were affected during in vitro conditions in comparison with AtqKNOX2. Conclusions Agave clones plants with higher DNA methylation during in vitro conditions were better adapted to ex vitro conditions. In addition

  3. Waste retrieval sluicing system data acquisition system acceptance test report

    International Nuclear Information System (INIS)

    Bevins, R.R.

    1998-01-01

    This document describes the test procedure for the Project W-320 Tank C-106 Sluicing Data Acquisition System (W-320 DAS). The Software Test portion will test items identified in the WRSS DAS System Description (SD), HNF-2115. Traceability to HNF-2115 will be via a reference that follows in parenthesis, after the test section title. The Field Test portion will test sensor operability, analog to digital conversion, and alarm setpoints for field instrumentation. The W-320 DAS supplies data to assist thermal modeling of tanks 241-C-106 and 241-AY-102. It is designed to be a central repository for information from sources that would otherwise have to be read, recorded, and integrated manually. Thus, completion of the DAS requires communication with several different data collection devices and output to a usable PC data formats. This test procedure will demonstrate that the DAS functions as required by the project requirements stated in Section 3 of the W-320 DAS System Description, HNF-2115

  4. Evolution of a test article handling system for the SP-100 GES test

    International Nuclear Information System (INIS)

    Shen, E.J.; Schweiger, L.J.; Miller, W.C.; Gluck, R.; Davies, S.M.

    1987-01-01

    A simulated space environment test of a flight prototypic SP-100 reactor, control system, and flight shield will be conducted at the Hanford Engineering Development Laboratory (HEDL). The flight prototypic components and the supporting primary heat removal system are collectively known as the nuclear assembly test article (TA). The unique configuration and materials of fabrication for the Test Article require a specialized handling facility to support installation, maintenance, and final disposal operation. The test site operator, working in conjunction with the test article supplier, developed and evaluated several handling concepts resulting in the selection of a reference test article handling system. The development of the reference concept for the handling system is presented

  5. Lightning vulnerability of nuclear explosive test systems at the Nevada Test Site

    International Nuclear Information System (INIS)

    Hasbrouck, R.T.

    1985-01-01

    A task force chartered to evaluate the effects of lightning on nuclear explosives at the Nevada Test Site has made several recommendations intended to provide lightning-invulnerable test device systems. When these recommendations have been implemented, the systems will be tested using full-threat-level simulated lightning

  6. The use of indium-111 platelet scintigraphy in man: comparisons with in vitro tests and in vivo platelet function--a five year experience

    International Nuclear Information System (INIS)

    Ezikowitz, M.D.; Ferri, P.; Pope, C.; Smith, E.O.; Snyder, E.L.

    1985-01-01

    In this paper, the authors present data collected from 540 patients between July 1978 and July 1983. They discuss briefly the method they employed for labeling platelets, emphasizing in vivo and in vitro markers of platelet function used for quality control of platelet preparation. They discuss the application of their technique for identification of mural left ventrical trombi, and review the disparity between in vivo and in vitro tests of platelet function. Then they deal with diagnosis of subacute bacterial endocarditis, deep veonus thrombosis, coronary trombi, left arterial masses, and the use of tomographic imaging. Finally they report changes in platelet function in stored platelets from normal volunteers

  7. An immunologic model for rapid vaccine assessment -- a clinical trial in a test tube.

    Science.gov (United States)

    Higbee, Russell G; Byers, Anthony M; Dhir, Vipra; Drake, Donald; Fahlenkamp, Heather G; Gangur, Jyoti; Kachurin, Anatoly; Kachurina, Olga; Leistritz, Del; Ma, Yifan; Mehta, Riyaz; Mishkin, Eric; Moser, Janice; Mosquera, Luis; Nguyen, Mike; Parkhill, Robert; Pawar, Santosh; Poisson, Louis; Sanchez-Schmitz, Guzman; Schanen, Brian; Singh, Inderpal; Song, Haifeng; Tapia, Tenekua; Warren, William; Wittman, Vaughan

    2009-09-01

    While the duration and size of human clinical trials may be difficult to reduce, there are several parameters in pre-clinical vaccine development that may be possible to further optimise. By increasing the accuracy of the models used for pre-clinical vaccine testing, it should be possible to increase the probability that any particular vaccine candidate will be successful in human trials. In addition, an improved model will allow the collection of increasingly more-informative data in pre-clinical tests, thus aiding the rational design and formulation of candidates entered into clinical evaluation. An acceleration and increase in sophistication of pre-clinical vaccine development will thus require the advent of more physiologically-accurate models of the human immune system, coupled with substantial advances in the mechanistic understanding of vaccine efficacy, achieved by using this model. We believe the best viable option available is to use human cells and/or tissues in a functional in vitro model of human physiology. Not only will this more accurately model human diseases, it will also eliminate any ethical, moral and scientific issues involved with use of live humans and animals. An in vitro model, termed "MIMIC" (Modular IMmune In vitro Construct), was designed and developed to reflect the human immune system in a well-based format. The MIMIC System is a laboratory-based methodology that replicates the human immune system response. It is highly automated, and can be used to simulate a clinical trial for a diverse population, without putting human subjects at risk. The MIMIC System uses the circulating immune cells of individual donors to recapitulate each individual human immune response by maintaining the autonomy of the donor. Thus, an in vitro test system has been created that is functionally equivalent to the donor's own immune system and is designed to respond in a similar manner to the in vivo response. 2009 FRAME.

  8. Harmonisation of animal testing alternatives in China.

    Science.gov (United States)

    Cheng, Shujun; Qu, Xiaoting; Qin, Yao

    2017-12-01

    More and more countries are lining up to follow the EU's approach and implement a full ban on the sale of cosmetics that have been tested on animals, which has been the case in the EU since 2013. Besides animal welfare considerations, the need for mutual acceptance of data (MAD) and harmonisation of the global market have made the move toward non-animal testing a desirable general trend for countries worldwide. Over the last 10 years, the concept of alternative methods has been gradually developing in China. This has seen the harmonisation of relevant legislation, the organisation of various theoretical and hands-on training sessions, the exploration of method validation, the adoption of internationally recognised methods, the propagation of alternative testing standards, and an in-depth investigation into the potential use of in vitro methods in the biosciences. There are barriers to this progress, including the demand for a completely new infrastructure, the need to build technology capability, the requirement for a national standardisation system formed through international co-operation, and the lack of technical assistance to facilitate self-innovation. China is now increasing speed in harmonising its approach to the use of non-animal alternatives, accelerating technological development and attempting to incorporate non-animal, in vitro, testing methods into the national regulatory system.

  9. Determining the optimal system-specific cut-off frequencies for filtering in-vitro upper extremity impact force and acceleration data by residual analysis.

    Science.gov (United States)

    Burkhart, Timothy A; Dunning, Cynthia E; Andrews, David M

    2011-10-13

    The fundamental nature of impact testing requires a cautious approach to signal processing, to minimize noise while preserving important signal information. However, few recommendations exist regarding the most suitable filter frequency cut-offs to achieve these goals. Therefore, the purpose of this investigation is twofold: to illustrate how residual analysis can be utilized to quantify optimal system-specific filter cut-off frequencies for force, moment, and acceleration data resulting from in-vitro upper extremity impacts, and to show how optimal cut-off frequencies can vary based on impact condition intensity. Eight human cadaver radii specimens were impacted with a pneumatic impact testing device at impact energies that increased from 20J, in 10J increments, until fracture occurred. The optimal filter cut-off frequency for pre-fracture and fracture trials was determined with a residual analysis performed on all force and acceleration waveforms. Force and acceleration data were filtered with a dual pass, 4th order Butterworth filter at each of 14 different cut-off values ranging from 60Hz to 1500Hz. Mean (SD) pre-fracture and fracture optimal cut-off frequencies for the force variables were 605.8 (82.7)Hz and 513.9 (79.5)Hz, respectively. Differences in the optimal cut-off frequency were also found between signals (e.g. Fx (medial-lateral), Fy (superior-inferior), Fz (anterior-posterior)) within the same test. These optimal cut-off frequencies do not universally agree with the recommendations of filtering all upper extremity impact data using a cut-off frequency of 600Hz. This highlights the importance of quantifying the filter frequency cut-offs specific to the instrumentation and experimental set-up. Improper digital filtering may lead to erroneous results and a lack of standardized approaches makes it difficult to compare findings of in-vitro dynamic testing between laboratories. Copyright © 2011 Elsevier Ltd. All rights reserved.

  10. Cooperative Testing of Uncontrollable Timed Systems

    DEFF Research Database (Denmark)

    David, Alexandre; Larsen, Kim Guldstrand; Li, Shuhao

    2008-01-01

    the SUT against the test purpose as long as the SUT reacts to our moves in a cooperative style. We present an operational framework of cooperative winning strategy generation, test case derivation and execution. The test method is proved to be sound and complete. Preliminary experimental results indicate......Abstract. This paper deals with targeted testing of timed systems with uncontrollable behavior. The testing activity is viewed as a game between the tester and the system under test (SUT) towards a given test purpose. The SUT is modeled as Timed Game Automaton and the test purpose is specified...... in Timed CTL formula. We can employ a timed game solver UPPAAL-TIGA to check if the test purpose is ture w.r.t. the model, and if yes, to generate a winning strategy and use it for black-box conformance testing. Specifically, we show that in case the checking yields a negative result, we can still test...

  11. Water electrolysis system refurbishment and testing

    Science.gov (United States)

    Greenough, B. M.

    1972-01-01

    The electrolytic oxygen generator for the back-up water electrolysis system in a 90-day manned test was refurbished, improved and subjected to a 182-day bench test. The performance of the system during the test demonstrated the soundness of the basic electrolysis concept, the high development status of the automatic controls which allowed completely hands-off operation, and the capability for orbital operation. Some design improvements are indicated.

  12. An in vitro method for detecting chemical sensitization using human reconstructed skin models and its applicability to cosmetic, pharmaceutical, and medical device safety testing.

    Science.gov (United States)

    McKim, James M; Keller, Donald J; Gorski, Joel R

    2012-12-01

    Chemical sensitization is a serious condition caused by small reactive molecules and is characterized by a delayed type hypersensitivity known as allergic contact dermatitis (ACD). Contact with these molecules via dermal exposure represent a significant concern for chemical manufacturers. Recent legislation in the EU has created the need to develop non-animal alternative methods for many routine safety studies including sensitization. Although most of the alternative research has focused on pure chemicals that possess reasonable solubility properties, it is important for any successful in vitro method to have the ability to test compounds with low aqueous solubility. This is especially true for the medical device industry where device extracts must be prepared in both polar and non-polar vehicles in order to evaluate chemical sensitization. The aim of this research was to demonstrate the functionality and applicability of the human reconstituted skin models (MatTek Epiderm(®) and SkinEthic RHE) as a test system for the evaluation of chemical sensitization and its potential use for medical device testing. In addition, the development of the human 3D skin model should allow the in vitro sensitization assay to be used for finished product testing in the personal care, cosmetics, and pharmaceutical industries. This approach combines solubility, chemical reactivity, cytotoxicity, and activation of the Nrf2/ARE expression pathway to identify and categorize chemical sensitizers. Known chemical sensitizers representing extreme/strong-, moderate-, weak-, and non-sensitizing potency categories were first evaluated in the skin models at six exposure concentrations ranging from 0.1 to 2500 µM for 24 h. The expression of eight Nrf2/ARE, one AhR/XRE and two Nrf1/MRE controlled gene were measured by qRT-PCR. The fold-induction at each exposure concentration was combined with reactivity and cytotoxicity data to determine the sensitization potential. The results demonstrated that

  13. The comet assay: assessment of in vitro and in vivo DNA damage.

    Science.gov (United States)

    Bajpayee, Mahima; Kumar, Ashutosh; Dhawan, Alok

    2013-01-01

    Rapid industrialization and pursuance of a better life have led to an increase in the amount of chemicals in the environment, which are deleterious to human health. Pesticides, automobile exhausts, and new chemical entities all add to air pollution and have an adverse effect on all living organisms including humans. Sensitive test systems are thus required for accurate hazard identification and risk assessment. The Comet assay has been used widely as a simple, rapid, and sensitive tool for assessment of DNA damage in single cells from both in vitro and in vivo sources as well as in humans. Already, the in vivo comet assay has gained importance as the preferred test for assessing DNA damage in animals for some international regulatory guidelines. The advantages of the in vivo comet assay are its ability to detect DNA damage in any tissue, despite having non-proliferating cells, and its sensitivity to detect genotoxicity. The recommendations from the international workshops held for the comet assay have resulted in establishment of guidelines. The in vitro comet assay conducted in cultured cells and cell lines can be used for screening large number of compounds and at very low concentrations. The in vitro assay has also been automated to provide a high-throughput screening method for new chemical entities, as well as environmental samples. This chapter details the in vitro comet assay using the 96-well plate and in vivo comet assay in multiple organs of the mouse.

  14. Characterization of DNA repair phenotypes of Xeroderma pigmentosum cell lines by a paralleled in vitro test

    International Nuclear Information System (INIS)

    Raffin, A.L.

    2009-06-01

    DNA is constantly damaged modifying the genetic information for which it encodes. Several cellular mechanisms as the Base Excision Repair (BER) and the Nucleotide Excision Repair (NER) allow recovering the right DNA sequence. The Xeroderma pigmentosum is a disease characterised by a deficiency in the NER pathway. The aim of this study was to propose an efficient and fast test for the diagnosis of this disease as an alternative to the currently available UDS test. DNA repair activities of XP cell lines were quantified using in vitro miniaturized and paralleled tests in order to establish DNA repair phenotypes of XPA and XPC deficient cells. The main advantage of the tests used in this study is the simultaneous measurement of excision or excision synthesis (ES) of several lesions by only one cellular extract. We showed on one hand that the relative ES of the different lesions depend strongly on the protein concentration of the nuclear extract tested. Working at high protein concentration allowed discriminating the XP phenotype versus the control one, whereas it was impossible under a certain concentration's threshold. On the other hand, while the UVB irradiation of control cells stimulated their repair activities, this effect was not observed in XP cells. This study brings new information on the XPA and XPC protein roles during BER and NER and underlines the complexity of the regulations of DNA repair processes. (author)

  15. Integration of QSAR and in vitro toxicology.

    Science.gov (United States)

    Barratt, M D

    1998-01-01

    The principles of quantitative structure-activity relationships (QSAR) are based on the premise that the properties of a chemical are implicit in its molecular structure. Therefore, if a mechanistic hypothesis can be proposed linking a group of related chemicals with a particular toxic end point, the hypothesis can be used to define relevant parameters to establish a QSAR. Ways in which QSAR and in vitro toxicology can complement each other in development of alternatives to live animal experiments are described and illustrated by examples from acute toxicological end points. Integration of QSAR and in vitro methods is examined in the context of assessing mechanistic competence and improving the design of in vitro assays and the development of prediction models. The nature of biological variability is explored together with its implications for the selection of sets of chemicals for test development, optimization, and validation. Methods are described to support the use of data from in vivo tests that do not meet today's stringent requirements of acceptability. Integration of QSAR and in vitro methods into strategic approaches for the replacement, reduction, and refinement of the use of animals is described with examples. PMID:9599692

  16. Efficacy of two rotary retreatment systems in removing Gutta-percha and sealer during endodontic retreatment with or without solvent: A comparative in vitro study.

    Science.gov (United States)

    Bhagavaldas, Moushmi Chalakkarayil; Diwan, Abhinav; Kusumvalli, S; Pasha, Shiraz; Devale, Madhuri; Chava, Deepak Chowdary

    2017-01-01

    The aim of this in vitro study was to compare the efficacy of two retreatment rotary systems in the removal of Gutta-percha (GP) and sealer from the root canal walls with or without solvent. Forty-eight extracted human mandibular first premolars were prepared and obturated with GP and AH Plus sealer. Samples were then randomly divided into four groups. Group I was retreated with MtwoR rotary system without solvent, Group II was retreated with MtwoR rotary system with Endosolv R as the solvent, Group III with D-RaCe rotary system without solvent, and Group IV with D-RaCe rotary system and Endosolv R solvent. The cleanliness of canal walls was determined by stereomicroscope (×20) and AutoCAD software. Kruskal-Wallis test and Mann-Whitney U-test were used to compare the data. Results showed that none of the retreatment systems used in this study was able to completely remove the root canal filling material. D-RaCe with or without solvent showed significantly ( P > 0.05) less filling material at all levels compared to MtwoR with/without solvent. Within the limitation of the current study, D-RaCe rotary retreatment system is more effective in removing filling material from root canal walls when compared to MtwoR rotary retreatment system.

  17. Induction of the acrosome reaction test to in vitro estimate embryo production in Nelore cattle

    Directory of Open Access Journals (Sweden)

    M.Z. Costa

    2010-08-01

    Full Text Available The effectiveness of induction of the acrosome reaction (AR test as a parameter to in vitro estimate embryo production (IVP in Nelore breed and the AR pattern by the Trypan Blue/Giemsa (TB stain were evaluated. Frozen semen samples from ten Nelore bulls were submitted to AR induction and were also evaluated for cleavage and blastocyst rates. The treatments utilized for AR induction were: control (TALP medium, TH (TALP medium + 10μg heparin, TL (TALP medium + 100μg lysophosphatidylcholine and THL (TALP medium + 10μg heparin + 100μg lysophosphatidylcholine. Sperm acrosomal status and viability were evaluated by TB staining at 0 and after 4h incubation at 38°C. The results obtained for AR presented a significant difference (P<0.05 in the percentage of acrosome reacted live sperm after 4h of incubation in the treatments that received heparin. The cleavage and blastocyst rates were 60% and 38% respectively and a significant difference was observed among bulls (P<0.05. It was founded a satisfactory model to estimate the cleavage and blastocyst rates by AR induction test. Therefore, it can be concluded that the induction of the AR test is a valuable tool to predict the IVP in Nelore breed.

  18. In vitro propagation of ‘FHIA-25’ (Musa spp., AAB in Temporary Immersion Systems

    Directory of Open Access Journals (Sweden)

    Milagros Basail Pérez

    2011-01-01

    Full Text Available Necessity to produce high quality planting material has required searching new alternatives to increase the efficiency of in vitro propagation methods and their automation, as in Temporary Immersion Systems (TIS. This work was aimed to multiply the hybrid ‘FHIA-25’ (Musa spp. AAB in TIS. The effect of different culture medium volumes per explant and densities of planting materials per culture flask at the same immersion frequency was determined. These two factors showed influence on the evaluated variables. A volume of 40 ml of culture medium per explant and densities of 80 explants per flask were selected to multiply this cultivar in TIS. These results permitted to increase in vitro production of high-quality plants for rooting stage. Key words: multiplication coefficient, culture flasks, liquid culture medium

  19. Stress Distribution in Single Dental Implant System: Three-Dimensional Finite Element Analysis Based on an In Vitro Experimental Model.

    Science.gov (United States)

    Rezende, Carlos Eduardo Edwards; Chase-Diaz, Melody; Costa, Max Doria; Albarracin, Max Laurent; Paschoeto, Gabriela; Sousa, Edson Antonio Capello; Rubo, José Henrique; Borges, Ana Flávia Sanches

    2015-10-01

    This study aimed to analyze the stress distribution in single implant system and to evaluate the compatibility of an in vitro model with finite element (FE) model. The in vitro model consisted of Brånemark implant; multiunit set abutment of 5 mm height; metal-ceramic screw-retained crown, and polyurethane simulating the bone. Deformations were recorded in the peri-implant region in the mesial and distal aspects, after an axial 300 N load application at the center of the occlusal aspect of the crown, using strain gauges. This in vitro model was scanned with micro CT to design a three-dimensional FE model and the strains in the peri-implant bone region were registered to check the compatibility between both models. The FE model was used to evaluate stress distribution in different parts of the system. The values obtained from the in vitro model (20-587 με) and the finite element analysis (81-588 με) showed agreement among them. The highest stresses because of axial and oblique load, respectively were 5.83 and 40 MPa for the cortical bone, 55 and 1200 MPa for the implant, and 80 and 470 MPa for the abutment screw. The FE method proved to be effective for evaluating the deformation around single implant. Oblique loads lead to higher stress concentrations.

  20. An introduction to testing techniques in the Intelsat TDMA/DSI system. II - Satellite system operations guide (SSOG) tests

    Science.gov (United States)

    Colby, R. J.; Parthasarathy, R.; Stimson, A. L.

    1983-12-01

    The test methods and procedures of the Intelsat TDMA/DSI SSOG are summarized. The overall structure of the SSOG is outlined, and the operational procedures to be followed for joining a new terminal to the system and for normal operations are reviewed, with an emphasis on the roles of the IOC and the TDMA reference and monitoring stations. The testing philosophy (based on minimal interruptions) and the star-test method are explained and illustrated with diagrams, and the test procedures are examined, including modem tests, electrical-path-length equalization, RF/IF downchain tests, protocol tests, nontransmitting protocol tests, IF/RF upchain tests, transmitting protocol tests, baseband tests, and orderwire lineups. The fundamental access discipline of the TDMA system is presented in an appendix.

  1. Comparison of two systems for rigidly connecting 2.0-mm bone screws to an implantable device : in vitro stability testing

    NARCIS (Netherlands)

    van Loon, JP; de Bont, LGM; Verkerke, GJ

    The stability of a screw-fixed implantable device can be improved by eliminating the freedom of movement between the screws and the device. Two systems have been developed for rigidly connecting 2.0-mm bone screws to an implantable device, and the aim of this study was to test and compare the

  2. Characterization of new eye drops with choline salicylate and assessment of their irritancy by in vitro short time exposure tests.

    Science.gov (United States)

    Wroblewska, Katarzyna; Kucinska, Małgorzata; Murias, Marek; Lulek, Janina

    2015-09-01

    The aim of our study was to examine the irritation potential of new eye drops containing 2% choline salicylate (CS) as an active pharmaceutical ingredient (API) and various polymers increasing eye drop viscosity (hydroxyethylcellulose, hydroxypropyl methylcellulose, methylcellulose, polyvinyl alcohol, polyvinylpyrrolidone). The standard method for assessing the potential of irritating substances has been the Draize rabbit eye test. However the European Centre for Validation of Alternative Methods and the Coordinating Committee for Validation of Alternative Methods recommend, short time exposure (STE) in vitro tests as an alternative method for assessing eye irritation. The eye irritation potential was determined using cytotoxicity test methods for rabbit corneal cell line (SIRC) after 5 min exposure. The viability of cells was determined using two cytotoxicity assays: MTT and Neutral Red Uptake. According to the irritation rankings for the short time exposure test, all tested eye drops are classified as non-irritating (cell viability >70%).

  3. Quality management systems for your in vitro fertilization clinic′s laboratory: Why bother?

    Directory of Open Access Journals (Sweden)

    Jan I Olofsson

    2013-01-01

    Full Text Available Several countries have in recent years introduced prescribed requirements for treatment and monitoring of outcomes, as well as a licensing or accreditation requirement for in vitro fertilization (IVF clinics and their laboratories. It is commonplace for Assisted Reproductive Technology (ART laboratories to be required to have a quality control system. However, more effective Total Quality Management systems are now being implemented by an increasing number of ART clinics. In India, it is now a requirement to have a quality management system in order to be accredited and to help meet customer demand for improved delivery of ART services. This review contains the proceedings a quality management session at the Indian Fertility Experts Meet (IFEM 2010 and focuses on the creation of a patient-oriented best-in-class IVF laboratory.

  4. Comparative antibacterial efficacies of hydrodynamic and ultrasonic irrigation systems in vitro.

    Science.gov (United States)

    Cachovan, Georg; Schiffner, Ulrich; Altenhof, Saskia; Guentsch, Arndt; Pfister, Wolfgang; Eick, Sigrun

    2013-09-01

    To ensure root canal treatment success, endodontic microbiota should be efficiently reduced. The in vitro bactericidal effects of a hydrodynamic system and a passive ultrasonic irrigation system were compared. Single-rooted extracted teeth (n = 250) were contaminated with suspensions of Enterococcus faecalis ATCC 29212, mixed aerobic cultures, or mixed anaerobic cultures. First, the antibacterial effects of the hydrodynamic system (RinsEndo), a passive ultrasonic irrigation system (Piezo smart), and manual rinsing with 0.9% NaCl (the control) were compared. Colony-forming units were counted. Second, the 2 systems were used with 1.5% sodium hypochlorite (NaOCl) alone or NaOCl + 0.2% chlorhexidine (CHX). The colony-forming units in the treated and untreated roots were determined during a period of 5 days. Both irrigation systems reduced bacterial numbers more effectively than manual rinsing (P irrigation reduced bacterial counts significantly better than hydrodynamic irrigation (P = .042). The NaOCl + CHX combination was more effective than NaOCl alone for both systems (P irrigation was more effective with NaOCl + CHX than the passive ultrasonic irrigation system. Both irrigation systems, when combined with NaOCl + CHX, removed bacteria from root canals. Copyright © 2013 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  5. In vitro recombination of bacteriophage T7 DNA damaged by uv radiation

    International Nuclear Information System (INIS)

    Masker, W.E.; Kuemmerle, N.B.

    1980-01-01

    A system capable of in vitro packaging of exogenous bacteriophage T7 DNA has been used to monitor the biological activity of DNA replicated in vitro. This system has been used to follow the effects of uv radiation on in vitro replication and recombination. During the in vitro replication process, a considerable exchange of genetic information occurs between T7 DNA molecules present in the reaction mixture. This in vitro recombination is reflected in the genotype of the T7 phage produced after in vitro encapsulation; depending on the genetic markers selected, recombinants can comprise nearly 20% of the total phage production. When uv-irradiated DNA is incubated in this system, the amount of in vitro synthesis is reduced and the total amount of viable phage produced after in vitro packaging is diminished. In vitro recombination rates are also lower when the participating DNA molecules have been exposed to uv. However, biochemical and genetic measurements confirmed that there is little or no transfer of pyrimidine dimers from irradiated DNA into undamaged molecules

  6. PFN tool test and calibration system

    International Nuclear Information System (INIS)

    Stephenson, W.A.

    1981-12-01

    A system has been developed for the functional testing and neutron output calibration of the PFN (Prompt Fission Neutron) Uranium Logging Tool. The system was designed primarily for field work and consists of a special vehicle as well as test apparatus. Only the pertinent instrumentation is described. This document will serve as an Instruction and Test Equipment service manual for those involved with calibration of the neutron output of the PFN tool

  7. Design, manufacture and in-vitro evaluation of a new microvascular anastomotic device.

    Science.gov (United States)

    Huang, Shao-Fu; Wang, Tien-Hsiang; Wang, Hsuan-Wen; Huang, Shu-Wei; Lin, Chun-Li; Kuo, Hsien-Nan; Yu, Tsung-Chih

    2013-01-01

    Many microvascular anastomoses have been proposed for use with physical assisted methods, such as cuff, ring-pin, stapler, clip to the anastomose blood vessel. The ring-pin type anastomotic device (e.g., 3M Microvascular Anastomotic System) is the most commonly used worldwide because the anastomotic procedure can be conducted more rapidly and with fewer traumas than using sutures. However, problems including vessel leakage, ring slippage, high cost and high surgical skill demand need to be resolved. The aim of this study is to design and manufacture a new anastomotic device for microvascular anastomosis surgery and validate the device functions with in-vitro testing. The new device includes one pair of pinned rings and a set of semi-automatic flap apparatus designed and made using computer-aided design / computer-aided manufacture program. A pair of pinned rings was used to impale vessel walls and establish fluid communication with rings joined. The semi-automatic flap apparatus was used to assist the surgeon to invert the vessel walls and impale onto each ring pin, then turning the apparatus knob to bring the rings together. The device was revised until it became acceptable for clinical requires. An in-vitro test was performed using a custom-made seepage micro-fluid system to detect the leakage of the anastomotic rings. The variation between input and output flow for microvascular anastomoses was evaluated. The new microvascular anastomotic device was convenient and easy to use. It requires less time than sutures to invert and impale vessel walls onto the pinned rings using the semi-automatic flap apparatus. The in-vitro test data showed that there were no tears from the joined rings seam during the procedures. The new anastomotic devices are effective even with some limitations still remaining. This device can be helpful to simplify the anastomosis procedure and reduce the surgery time.

  8. Automated System Tests High-Power MOSFET's

    Science.gov (United States)

    Huston, Steven W.; Wendt, Isabel O.

    1994-01-01

    Computer-controlled system tests metal-oxide/semiconductor field-effect transistors (MOSFET's) at high voltages and currents. Measures seven parameters characterizing performance of MOSFET, with view toward obtaining early indication MOSFET defective. Use of test system prior to installation of power MOSFET in high-power circuit saves time and money.

  9. Development and Application of Computational/In Vitro Toxicological Methods for Chemical Hazard Risk Reduction of New Materials for Advanced Weapon Systems

    Science.gov (United States)

    Frazier, John M.; Mattie, D. R.; Hussain, Saber; Pachter, Ruth; Boatz, Jerry; Hawkins, T. W.

    2000-01-01

    The development of quantitative structure-activity relationship (QSAR) is essential for reducing the chemical hazards of new weapon systems. The current collaboration between HEST (toxicology research and testing), MLPJ (computational chemistry) and PRS (computational chemistry, new propellant synthesis) is focusing R&D efforts on basic research goals that will rapidly transition to useful products for propellant development. Computational methods are being investigated that will assist in forecasting cellular toxicological end-points. Models developed from these chemical structure-toxicity relationships are useful for the prediction of the toxicological endpoints of new related compounds. Research is focusing on the evaluation tools to be used for the discovery of such relationships and the development of models of the mechanisms of action. Combinations of computational chemistry techniques, in vitro toxicity methods, and statistical correlations, will be employed to develop and explore potential predictive relationships; results for series of molecular systems that demonstrate the viability of this approach are reported. A number of hydrazine salts have been synthesized for evaluation. Computational chemistry methods are being used to elucidate the mechanism of action of these salts. Toxicity endpoints such as viability (LDH) and changes in enzyme activity (glutahoione peroxidase and catalase) are being experimentally measured as indicators of cellular damage. Extrapolation from computational/in vitro studies to human toxicity, is the ultimate goal. The product of this program will be a predictive tool to assist in the development of new, less toxic propellants.

  10. Test Confessions : A Study of Testing Practices for Plug-in Systems

    NARCIS (Netherlands)

    Greiler, M.; Van Deursen, A.; Storey, M.A.

    2011-01-01

    Testing plug-in-based systems is challenging due to complex interactions among many different plug-ins, and variations in version and configuration. The objective of this paper is to increase our understanding of what testers and developers think and do when it comes to testing plug-inbased systems.

  11. Root damage induced by intraosseous anesthesia. An in vitro investigation.

    Science.gov (United States)

    Graetz, Christian; Fawzy-El-Sayed, Karim-Mohamed; Graetz, Nicole; Dörfer, Christof-Edmund

    2013-01-01

    The principle of the intraosseous anesthesia (IOA) relies on the perforation of the cortical plate of the bone for direct application of the local anesthetic solution into the underlying cancellous structures. During this procedure, IOA needles might accidentally come in contact with the tooth roots. The aim of the current in vitro study was to examine the consequences of this 'worst case scenario' comparing five commercially available IOA systems. Extracted human roots were randomly perforated using five different IOA systems with a drilling time ≤5s. To simulate normal in vivo conditions, the roots were kept humid during the drilling procedure. Data was statistically evaluated using F-test (SPSS16, SPSS Inc., Chicago, USA) and the significance level was set at p ≤ 0.05. All examined systems resulted in root perforation. Drill fractures occurred in either none 0% (Quicksleeper, Anesto, Intraflow, Stabident) or 100% (X-Tip) of the applications. Excessive heat generation, as evident by combustion odor as well as metal and tooth discoloration, appeared in 30% (Quicksleeper), 40% (Anesto), 60% (Intraflow), 90% (Stabident) and 100% (X-Tip) of all perforations. Within the limits of in-vitro studies, the results show a potential for irreversible root damage that might be inflicted by an improper use of IOA systems.

  12. Mammalian oocyte growth and development in vitro.

    Science.gov (United States)

    Eppig, J J; O'Brien, M; Wigglesworth, K

    1996-06-01

    This paper is a review of the current status of technology for mammalian oocyte growth and development in vitro. It compares and contrasts the characteristics of the various culture systems that have been devised for the culture of either isolated preantral follicles or the oocyte-granulosa cell complexes form preantral follicles. The advantages and disadvantages of these various systems are discussed. Endpoints for the evaluation of oocyte development in vitro, including oocyte maturation and embryogenesis, are described. Considerations for the improvement of the culture systems are also presented. These include discussions of the possible effects of apoptosis and inappropriate differentiation of oocyte-associated granulosa cells on oocyte development. Finally, the potential applications of the technology for oocyte growth and development in vitro are discussed. For example, studies of oocyte development in vitro could help to identify specific molecules produced during oocyte development that are essential for normal early embryogenesis and perhaps recognize defects leading to infertility or abnormalities in embryonic development. Moreover, the culture systems may provide the methods necessary to enlarge the populations of valuable agricultural, pharmaceutical product-producing, and endangered animals, and to rescue the oocytes of women about to undergo clinical procedures that place oocytes at risk.

  13. Orion Ammonia Boiler System Preflight Test Preparations

    Science.gov (United States)

    Levitt, Julia L.

    2017-01-01

    The Environmental Controls and Life Support Systems (ECLSS) branch at Kennedy Space Center (KSC) is currently undergoing preparations for ground testing of the Orion Multi-Purpose Crew Vehicle (MPCV) to prepare its subsystems for EM-1 (Exploration Mission-1). EM-1, Orions second unmanned flight, is a three-week long lunar mission during which the vehicle will complete a 6-day retrograde lunar orbit before returning to Earth. This paper focuses on the work done during the authors 16-week internship with the Mechanical Engineering Branch of KSCs Engineering Directorate. The authors project involved assisting with the preparations for testing the Orion MPCVs ammonia boiler system. The purpose of the ammonia boiler system is to keep the spacecraft sufficiently cool during the reentry portion of its mission, from service module (SM) separation to post-landing. This system is critical for keeping both the spacecraft (avionics and electronics) and crew alive during reentry, thus a successful test of the system is essential to the success of EM-1. XXXX The author was able to draft a detailed outline of the procedure for the ammonia system functional test. More work will need to be done on the vehicle power-up and power-down portions of the procedure, but the ammonia system testing portion of the procedure is thorough and includes vehicle test configurations, vehicle commands, and GSE. The author was able to compile a substantial list of questions regarding the ammonia system functional test with the help of her mentors. A significant number of these questions were answered in the teleconferences with Lockheed Martin.

  14. Testing of nuclear air-cleaning systems

    International Nuclear Information System (INIS)

    Anon.

    1975-01-01

    A standard is presented which describes methods for field-testing nuclear power plant air cleaning systems. Included are specifications for visual inspection; duct and housing leak test; mounting frame pressure leak test; airflow capacity, distribution, and residence time tests; air-aerosol mixing uniformity test; in place leak test of HEPA filter banks; multiple sampling technique; in-place leak test of adsorber stage; laboratory testing of adsorbent; and duct heater performance test

  15. Test and control computer user's guide for a digital beam former test system

    Science.gov (United States)

    Alexovich, Robert E.; Mallasch, Paul G.

    1992-01-01

    A Digital Beam Former Test System was developed to determine the effects of noise, interferers and distortions, and digital implementations of beam forming as applied to the Tracking and Data Relay Satellite 2 (TDRS 2) architectures. The investigation of digital beam forming with application to TDRS 2 architectures, as described in TDRS 2 advanced concept design studies, was conducted by the NASA/Lewis Research Center for NASA/Goddard Space Flight Center. A Test and Control Computer (TCC) was used as the main controlling element of the digital Beam Former Test System. The Test and Control Computer User's Guide for a Digital Beam Former Test System provides an organized description of the Digital Beam Former Test System commands. It is written for users who wish to conduct tests of the Digital Beam forming Test processor using the TCC. The document describes the function, use, and syntax of the TCC commands available to the user while summarizing and demonstrating the use of the commands wtihin DOS batch files.

  16. Preoperational test report, recirculation ventilation systems

    Energy Technology Data Exchange (ETDEWEB)

    Clifton, F.T.

    1997-11-11

    This represents a preoperational test report for Recirculation Ventilation Systems, Project W-030. Project W-030 provides a ventilation upgrade for the four Aging Waste Facility tanks. The system provides vapor space cooling of tanks AY1O1, AY102, AZ1O1, AZ102 and supports the ability to exhaust air from each tank. Each system consists of a valved piping loop, a fan, condenser, and moisture separator; equipment is located inside each respective tank farm in its own hardened building. The tests verify correct system operation and correct indications displayed by the central Monitor and Control System.

  17. Preoperational test report, recirculation ventilation systems

    International Nuclear Information System (INIS)

    Clifton, F.T.

    1997-01-01

    This represents a preoperational test report for Recirculation Ventilation Systems, Project W-030. Project W-030 provides a ventilation upgrade for the four Aging Waste Facility tanks. The system provides vapor space cooling of tanks AY1O1, AY102, AZ1O1, AZ102 and supports the ability to exhaust air from each tank. Each system consists of a valved piping loop, a fan, condenser, and moisture separator; equipment is located inside each respective tank farm in its own hardened building. The tests verify correct system operation and correct indications displayed by the central Monitor and Control System

  18. In vitro activity of tigecycline and comparators against carbapenem-resistant Enterobacteriaceae in Africa-Middle East countries: TEST 2007-2012.

    Science.gov (United States)

    Renteria, M I; Biedenbach, D J; Bouchillon, S K; Hoban, D J; Raghubir, N; Sajben, P

    2014-09-01

    Multidrug-resistant (MDR) Enterobacteriaceae are an emerging concern for healthcare providers. Infections caused by MDR pathogens are associated with increased costs, length of hospital stay, and morbidity and mortality rates. Carbapenem-resistant Enterobacteriaceae (CRE) continue to increase, and infections with these organisms are observed worldwide not only as hospital-acquired infections but also as community-acquired infections. Increasing antimicrobial resistance dictates the need for continued surveillance studies of common and MDR pathogens. The Tigecycline Evaluation Surveillance Trial (TEST) examined the susceptibility of pathogens isolated in Africa and the Middle East from 2007 to 2012. A total of 4155 Enterobacteriaceae isolates were evaluated to determine the in vitro activity and changes in resistance patterns for tigecycline and comparators. Carbapenem resistance was found in 191 (4.6%) of the isolates tested. Klebsiella pneumoniae was the most common CRE (64.9%), followed by Enterobacter cloacae (14.1%) and Escherichia coli (9.9%). Tigecycline MIC 90 values (minimum inhibitory concentration required to inhibit 90% of the isolates) were 2μg/mL against all of these enteric species, with susceptibility rates of 96.8%, 92.6% and 100%, respectively. Tigecycline had in vitro activity against CRE, with a 95.3% susceptibility rate. Copyright © 2014 International Society for Chemotherapy of Infection and Cancer. Published by Elsevier Ltd. All rights reserved.

  19. Molecular ecotoxicology of nanosilver guided using in vitro prognosis

    DEFF Research Database (Denmark)

    Hayashi, Yuya; Heckmann, Lars-Henrik; Engelmann, Péter

    2012-01-01

    To study the molecular and cellular basis of silver nanoparticle (AgNP) toxicity, we here used a recently established in vitro model of earthworm coelomocytes in comparison to the conventional in vivo molecular ecotoxicology approach. Compared to the latter where the test organisms are exposed...... to NPs of interest held in an environmental matrix, in vitro models benefit from the ease of controlling exposure conditions in a defined set of biochemical milieus that NPs may encounter. The AgNPs tested in the present study originated from the same source, but to enhance the colloidal stability...... in the in vitro test media the NPs were pre-treated with serum proteins. In addition to physical characterisation of AgNPs, the active silver ion fraction was measured (in serum-supplemented cell culture medium and in soil pore-water). Using flow cytometry and atomic absorption spectrophotometry, we show...

  20. 21 CFR 862.1710 - Total triiodothyronine test system.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Total triiodothyronine test system. 862.1710... Systems § 862.1710 Total triiodothyronine test system. (a) Identification. A total triiodothyronine test system is a device intended to measure the hormone triiodothyronine in serum and plasma. Measurements...

  1. Spaceport Command and Control System Automation Testing

    Science.gov (United States)

    Hwang, Andrew

    2017-01-01

    The Spaceport Command and Control System (SCCS) is the National Aeronautics and Space Administrations (NASA) launch control system for the Orion capsule and Space Launch System, the next generation manned rocket currently in development. This large system requires high quality testing that will properly measure the capabilities of the system. Automating the test procedures would save the project time and money. Therefore, the Electrical Engineering Division at Kennedy Space Center (KSC) has recruited interns for the past two years to work alongside full-time engineers to develop these automated tests, as well as innovate upon the current automation process.

  2. Spaceport Command and Control System Automated Testing

    Science.gov (United States)

    Stein, Meriel

    2017-01-01

    The Spaceport Command and Control System (SCCS) is the National Aeronautics and Space Administrations (NASA) launch control system for the Orion capsule and Space Launch System, the next generation manned rocket currently in development. This large system requires high quality testing that will properly measure the capabilities of the system. Automating the test procedures would save the project time and money. Therefore, the Electrical Engineering Division at Kennedy Space Center (KSC) has recruited interns for the past two years to work alongside full-time engineers to develop these automated tests, as well as innovate upon the current automation process.

  3. Automatic test system of the photomultipliers

    International Nuclear Information System (INIS)

    Shiino, Kazuo; Kono, Koji; Ishii, Takanobu; Kasai, Seiji; Yamada, Sakue; Kitamura, Shoichi.

    1990-03-01

    A test system of R580 photomultipliers (PMTs) was constructed for the ZEUS experiment HERA. In this report, we will describe the general feature of the test system, each component of the setup, the procedure of the measurements, the data analyses and the results of the first 800 PMT measurements. (author)

  4. Molecular analysis of radiation-induced mutations in vitro

    International Nuclear Information System (INIS)

    Kronenberg, A.

    1996-01-01

    This review will focus on the nature of specific locus mutations detected in mammalian cells exposed in vitro to different types of ionizing radiations. Ionizing radiation has been shown to produce a wide variety of heritable alterations in DNA. These range from single base pair substitutions to stable loss or translocation of large portions of whole chromosomes. Data will be reviewed for certain test systems that reveal different mutation spectra. Techniques for the analysis of molecular alterations include applications of the polymerase chain reaction, some of which may be coupled with DNA sequence analysis, and a variety of hybridization-based techniques. The complexity of large scale rearrangements is approached with cytogenetic techniques including high resolution banding and various applications of the fluorescence in situ hybridization (FISH) technique. Radiation-induced mutant frequencies and mutation spectra are a function of the linkage constraints on the recovery of viable mutants for a given locus and test system. 44 refs

  5. Performance test results of mock-up test facility of HTTR hydrogen production system

    International Nuclear Information System (INIS)

    Ohashi, Hirofumi; Inaba, Yoshitomo; Nishihara, Tetsuo

    2004-01-01

    For the purpose to demonstrate effectiveness of high-temperature nuclear heat utilization, Japan Atomic Energy Research Institute has been developing a hydrogen production system and has planned to connect the hydrogen production system to High Temperature Engineering Test Reactor (HTTR). Prior to construction of a HTTR hydrogen production system, a mock-up test facility was constructed to investigate transient behavior of the hydrogen production system and to establish system controllability. The Mock-up test facility with a full-scale reaction tube is an approximately 1/30-scale model of the HTTR hydrogen production system and an electric heater is used as a heat source instead of a reactor. After its construction, a performance test of the test facility was carried out in the same pressure and temperature conditions as those of the HTTR hydrogen production system to investigate its performance such as hydrogen production ability, controllability and so on. It was confirmed that hydrogen was stably produced with a hot helium gas about 120m 3 /h, which satisfy the design value, and thermal disturbance of helium gas during the start-up could be mitigated within the design value by using a steam generator. The mock-up test of the HTTR hydrogen production system using this facility will continue until 2004. (author)

  6. Testing cooperative systems with the MARS simulator

    NARCIS (Netherlands)

    Netten, B.D.; Wedemeijer, H.

    2010-01-01

    The complexity of cooperative systems makes the use of high fidelity simulation essential in the development and testing of cooperative applications and their interactions with other cooperative systems. In SAFESPOT a simulator test bench is setup to test the safety margin applications running on

  7. Predicting the risk of developmental toxicity from in vitro assays

    International Nuclear Information System (INIS)

    Spielmann, Horst

    2005-01-01

    Reproductive toxicity refers to the adverse effects of a substance on any aspect of the reproductive cycle, including the impairment of reproductive function, the induction of adverse effects in the embryo, such as growth retardation, malformations, and death. Due to the complexity of the mammalian reproductive cycle, it is impossible to model the whole cycle in a single in vitro system in order to detect chemical effects on mammalian reproduction. However, the cycle can be broken down in its biological components which may be studied individually or in combination. This approach has the advantage that the target tissue/organ of a developmental toxicant can be identified. In specific areas of developmental toxicity, a number of useful and promising in vitro models are already available. The individual tests may be used as building blocks of a tiered testing strategy. So far, research has focused on developing and validating tests covering only a few components of the reproductive cycle, in particular organogenesis of the embryo, reflecting important concerns for teratogenic chemicals. During the last three decades, a number of established models and promising new developments have emerged that will be discussed, e.g. culture of mammalian embryos and embryonic cells and tissues and the use of embryonic stem cells

  8. ELECTROFORCE 3330 TEST SYSTEM

    Data.gov (United States)

    Federal Laboratory Consortium — The Bose Electroforce 3330 is a test system with an axial electromagnetic linear motor, a torsional motor, and an environmental chamber for high and low temperature...

  9. Initial in vitro testing of a paediatric continuous-flow total artificial heart.

    Science.gov (United States)

    Fukamachi, Kiyotaka; Karimov, Jamshid H; Horvath, David J; Sunagawa, Gengo; Byram, Nicole A; Kuban, Barry D; Moazami, Nader

    2018-06-01

    Mechanical circulatory support has become standard therapy for adult patients with end-stage heart failure; however, in paediatric patients with congenital heart disease, the options for chronic mechanical circulatory support are limited to paracorporeal devices or off-label use of devices intended for implantation in adults. Congenital heart disease and cardiomyopathy often involve both the left and right ventricles; in such cases, heart transplantation, a biventricular assist device or a total artificial heart is needed to adequately sustain both pulmonary and systemic circulations. We aimed to evaluate the in vitro performance of the initial prototype of our paediatric continuous-flow total artificial heart. The paediatric continuous-flow total artificial heart pump was downsized from the adult continuous-flow total artificial heart configuration by a scale factor of 0.70 (1/3 of total volume) to enable implantation in infants. System performance of this prototype was evaluated using the continuous-flow total artificial heart mock loop set to mimic paediatric circulation. We generated maps of pump performance and atrial pressure differences over a wide range of systemic vascular resistance/pulmonary vascular resistance and pump speeds. Performance data indicated left pump flow range of 0.4-4.7 l/min at 100 mmHg delta pressure. The left/right atrial pressure difference was maintained within ±5 mmHg with systemic vascular resistance/pulmonary vascular resistance ratios between 1.4 and 35, with/without pump speed modulation, verifying expected passive self-regulation of atrial pressure balance. The paediatric continuous-flow total artificial heart prototype met design requirements for self-regulation and performance; in vivo pump performance studies are ongoing.

  10. Effect of ionizing radiation on platelet function in vitro

    International Nuclear Information System (INIS)

    Kalovidouris, A.E.; Papayannis, A.G.

    1981-01-01

    The effect of ionizing radiation on platelet function was investigated in vitro. Platelet-rich plasma (300x10 9 /l) was irradiated with doses of 1, 4, 10, 20 and 50 Gy. Platelet function tests were performed on both irradiated and control (non-irradiated) platelet samples. The platelet function tests were (1) platelet aggregation by ADP (1, 2, 4 μmol final concentration), adrenaline and collagen, (2) ADP-release from platelets, (3) clot retraction and (4) platelet factor-3 availability. It was found that roentgen irradiation of platelets in vitro did not affect these platelet function tests. (Auth.)

  11. Model-based testing for embedded systems

    CERN Document Server

    Zander, Justyna; Mosterman, Pieter J

    2011-01-01

    What the experts have to say about Model-Based Testing for Embedded Systems: "This book is exactly what is needed at the exact right time in this fast-growing area. From its beginnings over 10 years ago of deriving tests from UML statecharts, model-based testing has matured into a topic with both breadth and depth. Testing embedded systems is a natural application of MBT, and this book hits the nail exactly on the head. Numerous topics are presented clearly, thoroughly, and concisely in this cutting-edge book. The authors are world-class leading experts in this area and teach us well-used

  12. 21 CFR 862.1670 - Sorbitol dehydrogenase test system.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Sorbitol dehydrogenase test system. 862.1670... Systems § 862.1670 Sorbitol dehydrogenase test system. (a) Identification. A sorbitol dehydrogenase test system is a device intended to measure the activity of the enzyme sorbitol dehydrogenase in serum...

  13. 21 CFR 862.1630 - Protein (fractionation) test system.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Protein (fractionation) test system. 862.1630... Systems § 862.1630 Protein (fractionation) test system. (a) Identification. A protein (fractionation) test system is a device intended to measure protein fractions in blood, urine, cerebrospinal fluid, and other...

  14. Model-Driven Test Generation of Distributed Systems

    Science.gov (United States)

    Easwaran, Arvind; Hall, Brendan; Schweiker, Kevin

    2012-01-01

    This report describes a novel test generation technique for distributed systems. Utilizing formal models and formal verification tools, spe cifically the Symbolic Analysis Laboratory (SAL) tool-suite from SRI, we present techniques to generate concurrent test vectors for distrib uted systems. These are initially explored within an informal test validation context and later extended to achieve full MC/DC coverage of the TTEthernet protocol operating within a system-centric context.

  15. Construction of test-bed system of voltage management system to ...

    African Journals Online (AJOL)

    Construction of test-bed system of voltage management system to apply physical power system. ... Journal of Fundamental and Applied Sciences ... system of voltage management system (VMS) in order to apply physical power system.

  16. In vitro comparison rate of dental root canal transportation using two single file systems on the simulated resin blocks

    Directory of Open Access Journals (Sweden)

    Mohammad Javad Etesami

    2016-07-01

    Full Text Available Background and Aims: Cleaning and shaping is one of the most important stages in endodontic treatment. Single-file systems save time and reduce the risk of transmission of pathogens. This in vitro study was aimed to compare the rate of canal transportation after the preparation of the stimulated resin root canal with two single-file systems, namely Waveone and Reciproc. Materials and Methods: Thirty stimulated resin root canal blocks with size 8/0. 02 K file were randomly divided into two study groups. The preparation in Group A and Group B was performed using Reciproc and Waveone files, respectively. Pre and post- preparation photographs were taken and the images were superimposed to evaluate the inner and outer wall’s curvature tendency at three points (apical, middle and coronal using AutoCad pragram. Data were analyzed using T-test. Results: Based on the results, the degree of transportation in the inner and outer walls of the canal was less at the level of 3 millimeters (P0.05. Conclusion: Waveone showed better performance in the middle third of canal and this system maybe recommended.

  17. Acclimatization of in Vitro-derived Dendrobium

    Directory of Open Access Journals (Sweden)

    Jaime A. Teixeira da Silva

    2017-05-01

    Full Text Available The successful ex vitro establishment of Dendrobium plantlets raised in vitro determines the quality of the end product (cut flowers or potted plants in commercial production for economic gain. When in vitro Dendrobium plantlets are transplanted from the culture room to greenhouse conditions, they may desiccate or wilt rapidly and can die as a result of changes in the environment, unless substantial precautions are taken to adapt plantlets to a new environment. The acclimatization of in vitro-grown Dendrobium plantlets to an ex vitro environment by gradually weaning them towards ambient relative humidity and light levels facilitates better survival of young and physiologically sensitive plantlets. Dendrobium plantlets raised in vitro must thus undergo a period of acclimatization or transitional development to correct anatomical abnormalities and to enhance their physiological performance to ensure survival under ex vitro conditions. The most common approach to improve the survival of Dendrobium plantlets upon transfer to an ex vitro environment is their gradual adaptation to that environment. Under such conditions, plants convert rapidly from a heterotrophic or photomixotrophic state to an autotrophic growth, develop a fully functional root system, and better control their stomatal and cuticular transpiration. Gradual adaptation is carried out in a greenhouse by decreasing relative humidity using fog or mist chambers and by increasing light intensity using shading techniques. This review details the acclimatization and ex vitro survival of Dendrobium plants produced in vitro. This advice is also useful for other orchids.

  18. Preoperational test report, raw water system

    Energy Technology Data Exchange (ETDEWEB)

    Clifton, F.T.

    1997-10-29

    This represents the preoperational test report for the Raw Water System, Project W-030. Project W-030 provides a ventilation upgrade for the four Aging Waste Facility tanks. The system supplies makeup water to the W-030 recirculation evaporative cooling towers for tanks AY1O1, AY102, AZ1O1, AZ102. The Raw Water pipe riser and associated strainer and valving is located in the W-030 diesel generator building. The tests verify correct system operation and correct indications displayed by the central Monitor and Control System.

  19. Preoperational test report, raw water system

    International Nuclear Information System (INIS)

    Clifton, F.T.

    1997-01-01

    This represents the preoperational test report for the Raw Water System, Project W-030. Project W-030 provides a ventilation upgrade for the four Aging Waste Facility tanks. The system supplies makeup water to the W-030 recirculation evaporative cooling towers for tanks AY1O1, AY102, AZ1O1, AZ102. The Raw Water pipe riser and associated strainer and valving is located in the W-030 diesel generator building. The tests verify correct system operation and correct indications displayed by the central Monitor and Control System

  20. The endo-lysosomal system of brain endothelial cells is influenced by astrocytes in vitro

    DEFF Research Database (Denmark)

    Toth, Andrea E; Siupka, Piotr; P Augustine, Thomas J

    2018-01-01

    Receptor- and adsorptive-mediated transport through brain endothelial cells (BEC) of the blood-brain barrier (BBB) involves a complex array of subcellular vesicular structures, the endo-lysosomal system. It consists of several types of vesicles, such as early, recycling, and late endosomes......, retromer-positive structures, and lysosomes. Since this system is important for receptor-mediated transcytosis of drugs across brain capillaries, our aim was to characterise the endo-lysosomal system in BEC with emphasis on their interactions with astrocytes. We used primary porcine BEC in monoculture....... Altogether, our data pin-point unique features of BEC trafficking network, essentially mapping the endo-lysosomal system of in vitro BBB models. Consequently, our findings constitute a valuable basis for planning the optimal route across the BBB when advancing drug delivery to the brain....

  1. The Transrapid test facility between system development and system application

    Energy Technology Data Exchange (ETDEWEB)

    Baur, L [MVP GmbH, Muenchen (Germany)

    1996-12-31

    In the development of a new rail technology, such as the magnetic levitation, there is - in contrast to the further development of the railway technology - not the possibility to use existing routes for the technical verification of the system technology until the application. Instead of this there are two possibilities: Cost-effective preliminary development on test beds and small test facilities up to a development stage which justifies the (relatively serious) risk of realising a service route early and to conclude the system trials and verification there; cost-intensive construction of a large-scale test facility which permits an application-related verification of all important system functions and thus creates the technical pre-requisites for a low-risk system application; The presentation deals with the technical requirements of the system at the test facility the challenges and chances linked to its realisation and adjustment to the rapidly progressing state-of-the-art and which this way opens up for a minimisation of the technical application risk. (orig./HW)

  2. High Frequency Plant Regeneration System from Transverse Thin Cell Layer Section of In vitro Derived ‘Nadia’ Ginger Microrhizome

    Directory of Open Access Journals (Sweden)

    Dikash Singh THINGBAIJAM

    2014-03-01

    Full Text Available An efficient and reproducible procedure is outlined for rapid in vitro multiplication of Zingiber officinale var. ‘Nadia’ through high frequency shoot proliferation from transverse thin cell layer (tTCL sections of in vitro derived microrhizome. In vitro derived microrhizome of size 500 μm in thickness was used as initial explants for induction of somatic embryos. Among the different phytohormones tested, tTCL explants shows maximum calli proliferation in medium containing 2 mg/L 2,4-Dichlorophenoxyacetic acid (88.30±0.11%. Reduced concentration of 2,4 Dichlorophenoxyacetic acid was supplemented with different cytokinins for regeneration of callus. Among the different medium tested, optimum redifferentiation of somatic embryos were observed in medium containing 0.2 mg/L 2,4 Dichlorophenoxyacetic acid and 6.0 mg/L BAP (141.08±0.25. Clump of regenerated plantlets were further subculture and transfer into microrhizome inducing medium containing high sucrose concentration (8%. Plantlets with well developed microrhizome were successfully acclimatized and eventually transferred to the field. The application of studying embryo section for regeneration of plants might be useful alternative to ginger improvement programme. Histological analysis showed formation of somatic embryos and regenerated adventitious shoot.

  3. Cyber-Physical Energy Systems Modeling, Test Specification, and Co-Simulation Based Testing

    DEFF Research Database (Denmark)

    van der Meer, A. A.; Palensky, P.; Heussen, Kai

    2017-01-01

    The gradual deployment of intelligent and coordinated devices in the electrical power system needs careful investigation of the interactions between the various domains involved. Especially due to the coupling between ICT and power systems a holistic approach for testing and validating is required....... Taking existing (quasi-) standardised smart grid system and test specification methods as a starting point, we are developing a holistic testing and validation approach that allows a very flexible way of assessing the system level aspects by various types of experiments (including virtual, real......, and mixed lab settings). This paper describes the formal holistic test case specification method and applies it to a particular co-simulation experimental setup. The various building blocks of such a simulation (i.e., FMI, mosaik, domain-specific simulation federates) are covered in more detail...

  4. Short term mutagenicity tests and their application to irradiated foods

    International Nuclear Information System (INIS)

    Phillips, B.J.; Elias, P.S.

    1980-01-01

    Although traditional long-term animal tests are likely to continue to be required, these are not only extremely costly but are coming more and more to be recognised as an imprecise and unsatisfactory method of testing the safety of irradiated foods for human consumption. It is therefore clearly advisable to include a selection of quicker and more direct testing methods in any toxicological assessment procedures. The International Project has therefore undertaken a study of the feasibility of using the newer systems for investigation of irradiated foodstuffs. Although some work in this field has already been carried out, some shortcomings in the published work can be identified which justify a more detailed and intensive research programme. As expected, little difficulty has been encountered in testing food by methods involving mammals, but considerable effort has been required to adapt in vitro systems. The use of enzymatic digestion in vitro to provide food samples for testing in mammalian cell cultures has never been attempted before and the procedures developed by the Project represent a positive contribution to methodology in this field. A series of foodstuffs is being tested by a wide spectrum of short-term tests and the first results are now being obtained. (orig./MG) [de

  5. Development in Assay Methods for in Vitro Antimalarial Drug Efficacy Testing: A Systematic Review

    Directory of Open Access Journals (Sweden)

    Shweta Sinha

    2017-10-01

    Full Text Available The emergence and spread of drug resistance are the major challenges in malaria eradication mission. Besides various strategies laid down by World Health Organization, such as vector management, source reduction, early case detection, prompt treatment, and development of new diagnostics and vaccines, nevertheless the need for new and efficacious drugs against malaria has become a critical priority on the global malaria research agenda. At several screening stages, millions of compounds are screened (1,000–2,000,000 compounds per screening campaign, before pre-clinical trials to select optimum lead. Carrying out in vitro screening of antimalarials is very difficult as different assay methods are subject to numerous sources of variability across different laboratories around the globe. Despite this, in vitro screening is an essential part of antimalarial drug development as it enables to resource various confounding factors such as host immune response and drug–drug interaction. Therefore, in this article, we try to illustrate the basic necessity behind in vitro study and how new methods are developed and subsequently adopted for high-throughput antimalarial drug screening and its application in achieving the next level of in vitro screening based on the current approaches (such as stem cells.

  6. Virtualization of System of Systems Test and Evaluation

    Science.gov (United States)

    2012-06-04

    computers and is the primary enabler for virtualization. 2. Virtualization System Elements Parmalee, Peterson , Tillman, & Hatfield (1972) outlined the...The work of Abu-Taieh and El Sheikh, based on the work of Balci (1994, 1995), and Balci et al. ( 1996 ), seeks to organize types of tests and to...and testing. In A. Dasso & A. Funes (Eds.), Verification, validation, and testing in software engineering (pp. 155–184). Hershey , PA: Idea Group

  7. Phosphite fertilisers as inhibitors of Hymenoscyphus fraxineus (anamorph Chalara fraxinea growth in tests in vitro

    Directory of Open Access Journals (Sweden)

    Tkaczyk Miłosz

    2017-03-01

    Full Text Available This study is designed to test the potential for reducing the growth of the mycelium of the fungus Hymenoscyphus fraxineus (anamorph Chalara fraxinea by using phosphite preparations at various concentrations in vitro. The study shows that adding pure phosphite to potato dextrose agar media inhibits the development of the fungus, but if the preparation is applied in the form of ammonium phosphite (Actifos, the growth of fungus will be accelerated. Probably the addition of nitrogen contained in the product Actifos has positive effect on the mycelial growth, but pure phosphite restricts its development. These studies are preliminary and only show the potential use of phosphite to reduce the development of H. fraxineus; however, to completely confirm its operation, further research is needed in this area.

  8. In Vitro Activities of New Antimicrobials against Nocardia brasiliensis

    Science.gov (United States)

    Vera-Cabrera, Lucio; Gonzalez, Eva; Choi, Sung H.; Welsh, Oliverio

    2004-01-01

    The in vitro sensitivities of 30 strains of Nocardia brasiliensis to DA-7867, gatifloxacin, moxifloxacin, and BMS-284756 (garenoxacin) were determined using the broth microdilution method. All N. brasiliensis strains were sensitive to these antimicrobials. The most active drug in vitro was DA-7867, with a MIC at which 90% of the isolates tested were inhibited of 0.03 μg/ml and a MIC at which 50% of the isolates tested were inhibited of 0.06 μg/ml. PMID:14742215

  9. Formulation of gastroretentive floating drug delivery system using hydrophilic polymers and its in vitro characterization

    Directory of Open Access Journals (Sweden)

    Venkata Srikanth Meka

    2014-04-01

    Full Text Available The aim of the present research is to formulate and evaluate the gastroretentive floating drug delivery system of antihypertensive drug, propranolol HCl. Gastroretentive floating tablets (GRFT were prepared by using a synthetic hydrophilic polymer polyethylene oxide of different grades such as PEO WSR N-12 K and PEO 18 NF as release retarding polymers and calcium carbonate as gas generating agent. The GRFT were compressed by direct compression strategy and the tablets were evaluated for physico-chemical properties, in vitro buoyancy, swelling studies, in vitro dissolution studies and release mechanism studies. From the dissolution and buoyancy studies, F 9 was selected as an optimized formulation. The optimized formulation followed zero order rate kinetics with non-Fickian diffusion mechanism. The optimized formulation was characterised with FTIR studies and observed no interaction between the drug and the polymers.

  10. Sims Prototype System 2 test results: Engineering analysis

    Science.gov (United States)

    1978-01-01

    The testing, problems encountered, and the results and conclusions obtained from tests performed on the IBM Prototype System, 2, solar hot water system, at the Marshall Space Flight Center Solar Test Facility was described. System 2 is a liquid, non draining solar energy system for supplying domestic hot water to single residences. The system consists of collectors, storage tank, heat exchanger, pumps and associated plumbing and controls.

  11. Specification and Test of Real-Time Systems

    DEFF Research Database (Denmark)

    Nielsen, Brian

    of the system, and a set of constraint patterns which describes and enforces the timing and synchronization constraints among components. We propose new techniques for automated black box conformance testing of real-time systems against densely timed speci cations. A test generator tool examines a specification......Distributed real-time computer based systems are very complex and intrinsically difficult to specify and implement correctly; in part this is caused by the overwhelming number of possible interactions between system components, but especially by a lack of adequate methods and tools to deal...... of the desired system behavior and generates the necessary test cases. A main problem is to construct a reasonably small test suite that can be executed within allotted resources, while having a high likelihood of detecting unknown errors. Our goal has been to treat the time dimension of this problem thoroughly...

  12. A listening test system for automative audio

    DEFF Research Database (Denmark)

    Bech, Søren; Gulbol, Mehmet-Ali; Martin, Geoff

    2005-01-01

    This paper describes two listening tests that were performed to provide initial validation of an auralisation system (see Part 1) to mimic the acoustics of a car interior. The validation is based on a comparison of results from an in-car listening test and another test using the auralisation system...... and recordings of the stimuli used for the in-car test. The music samples for the test were chosen from a database of various CODEC examples from a previous extensive ITU test to validate the ITU-R BS.1387-1 standard....

  13. Test procedure for boxed waste assay system

    International Nuclear Information System (INIS)

    Wachter, J.

    1994-01-01

    This document, prepared by Los Alamos National Laboratory's NMT-4 group, details the test methodology and requirements for Acceptance/Qualification testing of a Boxed Waste Assay System (BWAS) designed and constructed by Pajarito Scientific Corporation. Testing of the BWAS at the Plutonium Facility (TA55) at Los Alamos National Laboratory will be performed to ascertain system adherence to procurement specification requirements. The test program shall include demonstration of conveyor handling capabilities, gamma ray energy analysis, and imaging passive/active neutron accuracy and sensitivity. Integral to these functions is the system's embedded operating and data reduction software

  14. System maintenance test plan for the TWRS controlled baseline database system

    International Nuclear Information System (INIS)

    Spencer, S.G.

    1998-01-01

    TWRS [Tank Waste Remediation System] Controlled Baseline Database, formally known as the Performance Measurement Control System, is used to track and monitor TWRS project management baseline information. This document contains the maintenance testing approach for software testing of the TCBD system once SCR/PRs are implemented

  15. Antimalarial activity of plumbagin in vitro and in animal models.

    Science.gov (United States)

    Sumsakul, Wiriyaporn; Plengsuriyakarn, Tullayakorn; Chaijaroenkul, Wanna; Viyanant, Vithoon; Karbwang, Juntra; Na-Bangchang, Kesara

    2014-01-12

    Plumbagin is the major active constituent in several plants including Plumbago indica Linn. (root). This compound has been shown to exhibit a wide spectrum of biological and pharmacological activities. The present study aimed to evaluate the in vitro and in vivo antimalarial activity of plumbagin including its acute and subacute toxicity in mice. In vitro antimalarial activity of plumbagin against K1 and 3D7 Plasmodium falciparum clones were assessed using SYBR Green I based assay. In vivo antimalarial activity was investigated in Plasmodium berghei-infected mouse model (a 4-day suppressive test). Plumbagin exhibited promising antimalarial activity with in vitro IC50 (concentration that inhibits parasite growth to 50%) against 3D7 chloroquine-sensitive P. falciparum and K1 chloroquine-resistant P. falciparum clones of 580 (270-640) and 370 (270-490) nM, respectively. Toxicity testing indicated relatively low toxicity at the dose levels up to 100 (single oral dose) and 25 (daily doses for 14 days) mg/kg body weight for acute and subacute toxicity, respectively. Chloroquine exhibited the most potent antimalarial activity in mice infected with P. berghei ANKA strain with respect to its activity on the reduction of parasitaemia on day 4 and the prolongation of survival time. Plumbagin at the dose of 25 mg/kg body weight given for 4 days was safe and produced weak antimalarial activity. Chemical derivatization of the parent compound or preparation of modified formulation is required to improve its systemic bioavailability.

  16. Ravuconazole self-emulsifying delivery system: in vitro activity against Trypanosoma cruzi amastigotes and in vivo toxicity

    Directory of Open Access Journals (Sweden)

    Spósito PA

    2017-05-01

    Full Text Available Pollyanna Álvaro Spósito,1 Ana Lia Mazzeti,1,2 Caroline de Oliveira Faria,1 Julio A Urbina,3 Gwenaelle Pound-Lana,1 Maria Terezinha Bahia,2 Vanessa Furtado Mosqueira1 1Laboratory of Pharmaceutics and Nanotechnology Research, Pharmacy Department, School of Pharmacy, Universidade Federal de Ouro Preto, Minas Gerais, Brazil; 2Parasite Diseases Research Laboratory, NUPEB, Medical School, Universidade Federal de Ouro Preto, MG, Brazil; 3Venezuelan Institute for Scientific Research, Apartado, Caracas, Venezuela Abstract: Self-emulsifying drug delivery systems (SEDDSs are lipid-based anhydrous formulations composed of an isotropic mixture of oil, surfactant, and cosurfactants usually presented in gelatin capsules. Ravuconazole (Biopharmaceutics Classification System [BCS] Class II is a poorly water-soluble drug, and a SEDDS type IIIA was designed to deliver it in a predissolved state, improving dissolution in gastrointestinal fluids. After emulsification, the droplets had mean hydrodynamic diameters <250 nm, zeta potential values in the range of −45 mV to −57 mV, and showed no signs of ravuconazole precipitation. Asymmetric flow field-flow fractionation with dynamic and multiangle laser light scattering was used to characterize these formulations in terms of size distribution and homogeneity. The fractograms obtained at 37°C showed a polydisperse profile for all blank and ravuconazole–SEDDS formulations but no large aggregates. SEDDS increased ravuconazole in vitro dissolution extent and rate (20% compared to free drug (3% in 6 h. The in vivo toxicity of blank SEDDS comprising Labrasol® surfactant in different concentrations and preliminary safety tests in repeated-dose oral administration (20 days showed a dose-dependent Labrasol toxicity in healthy mice. Ravuconazole–SEDDS at low surfactant content (10%, v/v in Trypanosoma cruzi-infected mice was safe during the 20-day treatment. The anti-T. cruzi activity of free ravuconazole

  17. Safety Evaluation of Cosmetic Ingredients: In Vitro Opportunities for the Identification of Contact Allergens

    Directory of Open Access Journals (Sweden)

    Emanuela Corsini

    2014-03-01

    Full Text Available Irritant and allergic contact dermatitis are undesired side effects in the development of drugs and cosmetics as well as after contact with environmental or industrial chemicals. Over the last decades, a great deal of progress has been made in the development of alternative In vitro test to assess these issues. Driven by the 7th Amendment to the European Cosmetic Directive, the EU policy on chemicals (the registration, evaluation, authorization and restriction of chemicals (REACH system, the update of the European legislation on the protection of animals used in research, and emerging visions and strategies for predicting toxicity, in vitro methods are likely to play a major role in the near future. On 12 December 2013, the European Union Reference Laboratory for Alternatives to Animal Testing (EURL ECVAM, part of the European Commission Joint Research Centre published its Recommendation on the Direct Peptide Reactivity Assay (DPRA for skin sensitization, capable of distinguishing sensitizers from non-sensitizers. Other assays (i.e., KeratinoSens™ assay will follow shortly. While a number of methods are at various stages of development and use, currently it is not possible to rank chemicals for their sensitizing potency, an issue that is important for a full safety assessment. It is expected that a predictive method to totally replace animal testing will be in the form of a test battery comprising molecular, cell-based, and/or computational methods, the so-called “Integrated Approaches to Testing and Assessment”. This review aims to discuss the state-of-the-art in the field of in vitro assessment of contact sensitizers.

  18. Toward the establishment of standardized in vitro tests for lipid-based formulations. 2. The effect of bile salt concentration and drug loading on the performance of type I, II, IIIA, IIIB, and IV formulations during in vitro digestion.

    Science.gov (United States)

    Williams, Hywel D; Anby, Mette U; Sassene, Philip; Kleberg, Karen; Bakala-N'Goma, Jean-Claude; Calderone, Marilyn; Jannin, Vincent; Igonin, Annabel; Partheil, Anette; Marchaud, Delphine; Jule, Eduardo; Vertommen, Jan; Maio, Mario; Blundell, Ross; Benameur, Hassan; Carrière, Frédéric; Müllertz, Anette; Pouton, Colin W; Porter, Christopher J H

    2012-11-05

    The LFCS Consortium was established to develop standardized in vitro tests for lipid-based formulations (LBFs) and to examine the utility of these tests to probe the fundamental mechanisms that underlie LBF performance. In this publication, the impact of bile salt (sodium taurodeoxycholate, NaTDC) concentration and drug loading on the ability of a range of representative LBFs to generate and sustain drug solubilization and supersaturation during in vitro digestion testing has been explored and a common driver of the potential for drug precipitation identified. Danazol was used as a model poorly water-soluble drug throughout. In general, increasing NaTDC concentrations increased the digestion of the most lipophilic LBFs and promoted lipid (and drug) trafficking from poorly dispersed oil phases to the aqueous colloidal phase (AP(DIGEST)). High NaTDC concentrations showed some capacity to reduce drug precipitation, although, at NaTDC concentrations ≥3 mM, NaTDC effects on either digestion or drug solubilization were modest. In contrast, increasing drug load had a marked impact on drug solubilization. For LBFs containing long-chain lipids, drug precipitation was limited even at drug loads approaching saturation in the formulation and concentrations of solubilized drug in AP(DIGEST) increased with increased drug load. For LBFs containing medium-chain lipids, however, significant precipitation was evident, especially at higher drug loads. Across all formulations a remarkably consistent trend emerged such that the likelihood of precipitation was almost entirely dependent on the maximum supersaturation ratio (SR(M)) attained on initiation of digestion. SR(M) defines the supersaturation "pressure" in the system and is calculated from the maximum attainable concentration in the AP(DIGEST) (assuming zero precipitation), divided by the solubility of the drug in the colloidal phases formed post digestion. For LBFs where phase separation of oil phases did not occur, a

  19. Multiloop integral system test (MIST)

    International Nuclear Information System (INIS)

    Gloudemans, J.R.

    1989-07-01

    The multiloop integral system test (MIST) was part of a multiphase program started in 1983 to address small-break loss-of-coolant accidents (SBLOCAs) specific to Babcock and Wilcox-designed plants. MIST was sponsored by the US Nuclear Regulatory Commission, the Babcock and Wilcox Owners Group, the Electric Power Research Institute, and Babcock and Wilcox. The unique features of the Babcock and Wilcox design, specifically the hot leg U-bends and steam generators, prevented the use of existing integral system data or existing integral system facilities to address the thermal-hydraulic SBLOCA questions. MIST and two other supporting facilities were specifically designed and constructed for this program, and an existing facility -- the once-through integral system (OTIS) -- was also used. Data from MIST and the other facilities will be used to benchmark the adequacy of system codes, such as RELAP5 and TRAC, for predicting abnormal plant transients. The individual tests are described in detail in Volumes 2 through 8 and Volume 11, and are summarized in Volume 1. Inter-group comparisons are addressed in this document, Volume 9. These comparisons are grouped as follows: mapping versus SBLOCA transients, SBLOCA, pump effects, and the effects of noncondensible gases. Appendix A provides an index and description of the microfiched plots for each test, which are enclosed with the corresponding Volumes 2 through 8. 147 figs., 5 tabs

  20. Photo irradiation Systems for In-Vitro Cultured Cells Phototherapy and Photobiology Experiments

    International Nuclear Information System (INIS)

    Serrano Navarro, Joel; Morales Lopez, Orestes M.; Hernandez Quintanas, Luis F.; Lopez Silva, Y.; Fabila Bustos, Diego A.; De la Rosa Vazquez, Jose M.; Valor Reed, Alma; Stolik Isakina, Suren; Brodin, Patrik N.; Guha, Chandan; Tome, Wolfgang A.

    2016-01-01

    The increase in research and application of various phototherapy methods, especially photodynamic therapy (PDT) has created the need to study in depth the mechanisms of interaction of light with biological tissue using a photosensitizing drug in order to increase the therapeutic effectiveness. In this issue, two systems for controlled irradiation of in-vitro cell culture and temperature monitoring of the culture are presented. The first system was designed to irradiate 24 wells in a 96-well microplate. The second one was constructed for the irradiation and control of a 24-well microplate using larger volumes of cultured cells. Both systems can independently irradiate and control the temperature of each well. The systems include a module for contactless measurement of the temperature in each well. Light sources are located in an interchangeable module, so that it can be replaced to irradiate with different wavelengths. These prototypes count with various operation modes, controlled by a computer, which permits establishing specific settings in accordance with the desired experiment. The systems allow the automated experiment execution with precise control of dosimetry, irradiation and temperature, which reduces the sample-handling while, saves time. (Author)