bluuebunny/crossref_metadata_2025 huggingface dataset:
Prioritisation of Polybrominated Diphenyl Ethers (PBDEs) by Using the QSPR-THESAURUS Web Tool
Authors: Tetko Igor V. , Sopasakis Pantelis , Kunwar Prakash , Brandmaier Stefan , Novoratskyi Sergii , Charochkina Larysa , Prokopenko Volodymyr , Peijnenburg Willie J.G.M.
Published: April 2019

The prioritisation of chemical compounds is important for the identification of those chemicals that represent the highest threat to the environment. As part of the CADASTER project ( http://www.cadaster.eu ), we developed an online web tool that allows the calculation of the environmental risk of chemical compounds from a web interface. The environmental fate of compounds in the aquatic compartment is assessed by using the SimpleBox model, while adverse effects on the aquatic compartment are assessed by the Species Sensitivity Distribution approach. The main purpose of this web tool is to exemplify the use of quantitative structure–activity relationships (QSARs) to support risk assessment. A case study of QSAR integrated risk assessment of 209 polybrominated diphenyl ethers (PBDEs) demonstrates the treatment and influence of uncertainty in the predicted physicochemical and toxicity parameters in probabilistic risk assessment.
The Ethics of Using Laboratory Animals to Develop Medicines for Lifestyle Diseases
Authors: Phillips Clive
Published: April 2019

Humans advance through life with their accolade of food and laboratory animals to sustain them, with the former unfortunately increasing the need for the latter
The Increasing Use of Genetically-altered Animals Threatens the Credibility of Directive 2010/63/EU
Authors: Balls Michael
Published: April 2019

The new EU Directive and the equivalent legislation in the EU Member States will be of little value, unless they can bring the production and use of genetically-altered animals under control and lead to a progressive and sustained reduction in laboratory animal use
Fish as Research Tools: Alternatives to In Vivo Experiments
Authors: Schaeck Marlien , Van den Broeck Wim , Hermans Katleen , Decostere Annemie
Published: April 2019

The use of fish in scientific research is increasing worldwide, due to both the rapid expansion of the fish farming industry and growing awareness of questions concerning the humane use of mammalian models in basic research and chemical testing. As fish are lower on the evolutionary scale than mammals, they are considered to be less sentient. Fish models are providing researchers, and those concerned with animal welfare, with opportunities for adhering to the Three Rs principles of refinement, reduction and replacement. However, it should be kept in mind that fish should also be covered by the principles of the Three Rs. Indeed, various studies have shown that fish are capable of nociception, and of experiencing pain in a manner analogous to that in mammals. Thus, emphasis needs to be placed on the development of alternatives that replace, as much as possible, the use of all living vertebrate animals, including fish. This review gives the first comprehensive and critical overview of the existing alternatives for live fish experimental studies. The alternative methods described range from cell and tissue cultures, organ and perfusion models, and embryonic models, to in silico computer and mathematical models. This article aspires to guide scientists in the adoption of the correct alternative methods in their research, and, whenever possible, to reduce the use of live fish.
6. Replacement
Authors: Balls Michael
Published: April 2019

The total replacement of animal procedures should be our common goal, not least because animal models cannot tell us what we need to know about human conditions and responses
The Three Rs — Do We Need a New Principle?
Authors: Wittevrongel Christiaan
Published: April 2019

A new generation of Russells and Burches are needed, who will bring in a new era of biological science that will truly benefit animals as well as humans
Animal Experimentation: Transparency and Openness Mean Little, Unless Accompanied by Honesty and Accountability
Authors: Balls Michael
Published: April 2019

Delivery Plans, and Declarations and Concordats on transparency and openness, are all very well, but what will they really achieve with respect to the Three Rs?
Our Guiding Principles
Authors: van Ekert Onay Katherine
Published: April 2019

Veterinary surgeons should let themselves be guided by the principle “In dubio pro animale” — roughly translated as “When in doubt, support the animals”
13. Rex Leonard Burch: Humane Scientist and Gentle Man
Authors: Balls Michael
Published: April 2019

Rex Burch played a vital role in collecting background information about practical animal experimentation, and was the first person to use the term ‘alternatives’ in that context. The senior author of The Principles, William Russell, greatly valued their collaboration
A Message to the Proponents of Animal Experimentation: The Truth Shall Make you Free
Authors: Balls Michael
Published: April 2019

Some thoughts provoked by a ruling by the Advertising Standards Authority
Toxicity Assessment of Tobacco Products in Vitro
Authors: Manuppello Joseph R. , Sullivan Kristie M.
Published: April 2019

Driven by new regulatory demands to demonstrate risk reduction, the toxicity assessment of tobacco products increasingly employs innovative in vitro methods, including biphasic cell and tissue cultures exposed to whole cigarette smoke at the air–liquid interface, cell transformation assays, and genomic analyses. At the same time, novel tobacco products are increasingly compared to traditional cigarettes. This overview of in vitro toxicology studies of tobacco products reported in the last five years provides evidence to support the prioritisation of in vitro over in vivo methods by industry and their recommendation by regulatory authorities.
Coffee in Class: An Alternative to Animal Experiments in Pharmacology?
Authors: Agarwal Anoop Kumar , Shehnaz Syed Ilyas , Khanam Razia , Arifulla Mohamed
Published: April 2019

The stimulant effect of coffee on psychomotor performance was introduced as a potential alternative clinical pharmacology experiment for medical and pharmacy students
3-D Tissue Modelling and Virtual Pathology as New Approaches to Study Ductal Carcinoma In Situ
Authors: Booth Mary E. , Nash Claire E. , Roberts Nicholas P. , Magee Derek R. , Treanor Darren , Hanby Andrew M. , Speirs Valerie
Published: April 2019

Widespread screening mammography programmes mean that ductal carcinoma in situ (DCIS), a pre-invasive breast lesion, is now more frequently diagnosed. However, not all diagnosed DCIS lesions progress to invasive breast cancer, which presents a dilemma for clinicians. As such, there is much interest in studying DCIS in the laboratory, in order to help understand more about its biology and determine the characteristics of those that progress to invasion. Greater knowledge would lead to targeted and better DCIS treatment. Here, we outline some of the models available to study DCIS, with a particular focus on animal-free systems.
First Thoughts on the Effects on the Protection of Laboratory Animals of the UK's Departure from the European Union
Authors: Balls Michael
Published: April 2019

Directive 2010/63/EU led to Improvements to the Animals (Scientific Procedures) Act 1986: it is Hoped that these Improvements will be Maintained, and that Special Attention will be paid to the use of Non-Human Primates and Dogs
Exploring Waiving Opportunities for Mammalian Acute Systemic Toxicity Tests
Authors: Graepel Rabea , Asturiol David , Prieto Pilar , Worth Andrew P.
Published: April 2019

A survey was carried out to explore opportunities for waiving mammalian acute systemic toxicity tests. We were interested in finding out whether data from a sub-acute toxicity test could be used to predict the outcome of an acute systemic toxicity test. The survey was directed at experts in the field of toxicity testing, and was carried out in the context of the upcoming 2018 final registration deadline for chemicals under the EU REACH Regulation. In addition to the survey, a retrospective data analysis of chemicals that had already been registered with the European Chemicals Agency, and for which both acute and sub-acute toxicity data were available, was carried out. This data analysis was focused on chemicals that were administered via the oral route. The answers to the questionnaire showed a willingness to adopt waiving opportunities. In addition, the responses showed that data from a sub-acute toxicity test or dose-range finding study might be useful for predicting chemicals that do not require classification for acute oral toxicity (LD50 > 2000mg/kg body weight). However, with the exception of substances that fall into the non-classified category, it is difficult to predict current acute oral toxicity categories.
The 2015 Lush Prize Awards
Authors: BéruBé Kelly A. , Redmond Craig
Published: April 2019

This year saw a Black Box Prize award, for definitive work on the Adverse Outcome Pathway for skin sensitisation
Body-on-a-Chip Systems for Animal-free Toxicity Testing
Authors: Mahler Gretchen J. , Esch Mandy B. , Stokol Tracy , Hickman James J. , Shuler Michael L.
Published: April 2019

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 pro-drug 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.
2015 Lush Science Prize
Authors: McCann Jenny , McCann Terry
Published: April 2019

The Lush Prize supports animal-free testing by rewarding the most effective projects and individuals who have been working toward the goal of replacing animals in product or ingredient safety testing. Prizes are awarded for developments in five strategic areas: Science; Lobbying; Training; Public Awareness; and Young Researchers. Should there be a major breakthrough in 21st century toxicology, a Black Box Prize equivalent to the entire annual fund of £250,000 is awarded. A Background Paper is prepared each year, prior to the judging process, to provide the panel with a brief overview of current developments in the field of Replacement alternatives, particularly those relevant to the concept of toxicity pathways. The Background Paper includes information on recent work by the relevant scientific institutions and projects in this area, including AXLR8, the OECD, The Hamner Institutes, the Human Toxome Project, EURL ECVAM, ICCVAM, the US Tox21 Programme, the ToxCast programme, and the Human Toxicology Project Consortium. Recent developments in toxicity pathway research are also assessed by reviewing the relevant literature (including conference proceedings), and the abstracts and papers receiving the highest score are presented to the judges for consideration.
Sample Size Estimation for Pilot Animal Experiments by Using a Markov Chain Monte Carlo Approach
Authors: Allgoewer Andreas , Mayer Benjamin
Published: April 2019

The statistical determination of sample size is mandatory when planning animal experiments, but it is usually difficult to implement appropriately. The main reason is that prior information is hardly ever available, so the assumptions made cannot be verified reliably. This is especially true for pilot experiments. Statistical simulation might help in these situations. We used a Markov Chain Monte Carlo (MCMC) approach to verify the pragmatic assumptions made on different distribution parameters used for power and sample size calculations in animal experiments. Binomial and normal distributions, which are the most frequent distributions in practice, were simulated for categorical and continuous endpoints, respectively. The simulations showed that the common practice of using five or six animals per group for continuous endpoints is reasonable. Even in the case of small effect sizes, the statistical power would be sufficiently large (≥ 80%). For categorical outcomes, group sizes should never be under eight animals, otherwise a sufficient statistical power cannot be guaranteed. This applies even in the case of large effects. The MCMC approach demonstrated to be a useful method for calculating sample size in animal studies that lack prior data. Of course, the simulation results particularly depend on the assumptions made with regard to the distributional properties and effects to be detected, but the same also holds in situations where prior data are available. MCMC is therefore a promising approach toward the more informed planning of pilot research experiments involving the use of animals.
Evaluation of a Human Neural Stem Cell Culture Method for Prediction of the Neurotoxicity of Anti-epileptics
Authors: Al-Rubai Abdal-jabbar , Wigmore Peter , Pratten Margaret K.
Published: April 2019

Human neural stem cells have been proposed as an in vitro model to predict neurotoxicity. In this study, the potential of in vitro cultures of human-derived neurospheres to predict the effects of various anti-epileptic drugs (sodium valproate, phenytoin, carbamazepine and phenobarbitone) was evaluated. In general, these drugs had no significant effects on cell viability, total cellular protein, and neuronal process length at low doses, but at high doses these parameters were reduced significantly. Therapeutic doses of sodium valproate and phenytoin had a clear effect on neurosphere size and cell migration, with a significant reduction in both parameters when compared with the control group. The other drugs (carbamazepine and phenobarbitone) reduced neurosphere size and cell migration only at higher doses. The expression levels of glial fibrillary protein and tubulin III, which were used to identify astrocytes and neuronal cells, respectively, were reduced in a dose-dependent manner that became significant at high doses. The levels of glial fibrillary protein did not indicate any occurrence of reactive astrocytosis.
Animal Experiments and the Right to Ask Questions about Them
Authors: Balls Michael
Published: April 2019

The 2016 Home Office Statistics reveal little about progress in implementing the Three Rs, and the right to question how laboratory animals are used in Great Britain has been challenged
Animal ‘Models': How a Mechanistic Approach can Reduce Suffering and Improve Translatability
Authors: Lilley Elliot
Published: April 2019

Poorly predictive animal models of disease cause avoidable suffering and hamper the discovery of new treatments for patients. A focus on mechanistic modelling has the potential to reduce animal suffering as well as improving translation from the bench to the bedside.
The Replacement of Animal Tests
Authors: Combes Robert D.
Published: April 2019

Progress toward the acceptance and application of validated alternative test methods as replacements for animal tests, is being frustrated by the unsatisfactory procedures involved in approving new test guidelines and deleting existing ones
Exploring the Use of Alternatives to Animals in Undergraduate Education in Australia
Authors: Mallia Catherine , Logan Patricia , Freire Rafael
Published: April 2019

The replacement, reduction and refinement of animal use in education is part of the regulatory legislation in Australia, and requires the use of alternatives to animals where appropriate. The aims of this study were: a) to explore the extent of the replacement of animals when teaching life sciences to Australian undergraduate students; b) to understand which alternative models were being used, and the learning objectives covered; and c) to gain some insight into the circumstances facilitating the use of alternatives to animals in education. An anonymous online survey, consisting of open and closed questions, was conducted among faculty members that used either animal or alternative models in their teaching. A total of 27 faculty members participated, from eight universities. Human anatomy and physiology had the highest number of survey participants who had replaced animals entirely with alternative models. These subjects also had the highest number of participants that were using animal models. According to the participants, most learning objectives were met effectively by both types of model. Participants who only used alternatives were influenced by ethical considerations significantly more than those who used animal models and alternatives. We concluded that, while some participants have replaced animals successfully, others in the same field are still employing them, and that there appears to be a range of barriers to the wider adoption of alternatives to animal use.
Phase 0, Including Microdosing Approaches: Applying the Three Rs and Increasing the Efficiency of Human Drug Development
Authors: Burt Tal , Vuong Le Thuy , Baker Elizabeth , Young Graeme C. , McCartt A. Daniel , Bergstrom Mats , Sugiyama Yuichi , Combes Robert
Published: April 2019

Phase 0 approaches, including microdosing, involve the use of sub-therapeutic exposures to the tested drugs, thus enabling safer, more-relevant, quicker and cheaper first-in-human (FIH) testing. These approaches also have considerable potential to limit the use of animals in human drug development. Recent years have witnessed progress in applications, methodology, operations, and drug development culture. Advances in applications saw an expansion in therapeutic areas, developmental scenarios and scientific objectives, in, for example, protein drug development and paediatric drug development. In the operational area, the increased sensitivity of Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS), expansion of the utility of Positron Emission Tomography (PET) imaging, and the introduction of Cavity Ring-Down Spectroscopy (CRDS), have led to the increased accessibility and utility of Phase 0 approaches, while reducing costs and exposure to radioactivity. PET has extended the application of microdosing, from its use as a predominant tool to record pharmacokinetics, to a method for recording target expression and target engagement, as well as cellular and tissue responses. Advances in methodology include adaptive Phase 0/Phase 1 designs, cassette and cocktail microdosing, and Intra-Target Microdosing (ITM), as well as novel modelling opportunities and simulations. Importantly, these methodologies increase the predictive power of extrapolation from microdose to therapeutic level exposures. However, possibly the most challenging domain in which progress has been made, is the culture of drug development. One of the main potential values of Phase 0 approaches is the opportunity to terminate development early, thus not only applying the principle of ‘kill-early-kill-cheap’ to enhance the efficiency of drug development, but also obviating the need for the full package of animal testing required for therapeutic level Phase 1 studies. Finally, we list developmental scenarios that utilised Phase 0 approaches in novel drug development.
Selective Induction of Interleukin-12 in Reconstructed Human Epidermis by Chemical Allergens
Authors: Corsini Emanuela , Limiroli Elena , Marinovich Marina , Cohen Catherine , Roguet Roland , Galli Corrado L.
Published: April 2019

Keratinocytes play an important role in skin inflammatory and immunological reactions through the release of cytokines and response to them. These cells have been shown to direct T-cell priming by producing cytokines such as interleukin (IL)-10 and IL-12. The purpose of this work was to explore the potential use of IL-12 production to discriminate between skin irritants and contact allergens in vitro. Initially, a reconstituted human epidermis was treated with a known human skin irritant, sodium lauryl sulphate (SLS), and a known human contact allergen, 1-chloro-2,4-dinitrobenzene (DNCB). The expression of IL-12p40 was assessed at specific time intervals by the semi-quantitative reverse transcriptase-polymerase chain reaction (rt-PCR). The data obtained indicated that only DNCB induced an up-regulation of IL-12p40. This up-regulation occurred after exposure to DNCB for 3 hours. Importantly, the application of SLS or vehicles did not induce IL-12 mRNA up-regulation. An increase in total IL-12 protein content was detected in supernatants of allergen-stimulated, but not vehicle-stimulated, reconstituted epidermis. To confirm these results, the effects of benzalkonium chloride, oxazolone and eugenol were assessed. At concentrations that resulted in equivalent IL-1α release, only contact allergens increased IL-12 expression, which confirmed the previous results. These data suggest that IL-12, which is crucial for T-helper type 1 cell responses, could be a useful marker for discriminating between contact allergens and irritants.
Alternative Methods for Assessing Biocompatibility and Function of Implant Materials
Authors: Lewandowska-Szumiel Malgorzata
Published: April 2019

Biocompatibility testing is used to evaluate the host response to implantable materials and to assess their ability to perform in applications in which they are intended to interact with biological systems. In compliance with international and/or national standards, such assessment is based mainly on the results of experimental implantation into animal tissues. However, the development of in vitro experimental techniques creates new opportunities to observe and to understand the interaction of biomaterials with host tissue. The state-of-the-art application of alternative methods in biocompatibility testing is presented in this review article. It is discussed with respect to the Three Rs concept (reduction, refinement, replacement) of Russell & Burch. Perspectives on alternative methods in biocompatibility studies are discussed with regard to the possible role of biomaterials in tissue engineering.
Lack of Effect of Medium Supplementation with Pyruvate and Hormones on Cytochrome P450-mediated Activity of Rat Hepatocytes in Primary Culture
Authors: Wirachwong Porntip , Fry Jeffrey R.
Published: April 2019

The loss of cytochrome P450 (CYP)-dependent activity continues to be a problem in the use of cultured hepatocytes in xenobiotic toxicity studies. It has been reported that the inclusion of pyruvate and various hormones in the culture medium improves the maintenance of various hepatic functions, including that of CYP2C11 mRNA expression. We have studied this further, by investigating the effects of the addition of pyruvate and hormones on various CYP-dependent enzyme activities and on the CYP-dependent toxicity of precocene II in rat hepatocyte cultures. No beneficial effects of this medium supplementation could be demonstrated.
Establishment of an Embryotoxicity Assay with Green Fluorescence Protein-expressing Embryonic Cell-derived Cardiomyocytes
Authors: Bremer Susanne , Van Dooren Maaike , Paparella Martin , Kossolov Eugen , Fleischmann Bernd , Hescheler Juergen
Published: April 2019

Transgenic embryonic stem cells were used to determine the embryotoxic effects of chemicals on the development of embryonic tissues. This investigation supports an ongoing validation study, aimed at reducing the time-consuming procedure currently in use, and at providing more-objective and more-detailed information on the embryotoxic potentials of chemicals. Green fluorescence protein (GFP) was used as a reporter gene and was linked to a human α-cardiac-specific promoter. The expression of GFP was switched on after specific activation of the human α-actin promoter. This permitted the easy quantification of cardiac cells by using a fluorescence-activated cell sorter (FACS). The percentage of cardiac precursor cells was calculated from the FACS-distribution pattern of cells which fluoresced versus the total number of cells. The percentage of cardiac precursor cells increased from 25% in embryoid bodies on day 3, to 86% on day 7. However, in 11-day-old embryoid bodies, the percentage decreased to 35%. Five chemicals with known embryotoxic potentials were compared with respect to the IC50 (concentration causing 50% inhibition of measured effect) values obtained by various in vitro endpoints (for example, cytotoxicity, morphology). The results showed a higher sensitivity of endpoints used for the analysis of specific effects on the selected target tissue. The data also showed the need to develop in vitro methods with specific endpoints which account for the complexity of embryotoxicology.
Evaluation of a Short-term Rat Proximal Tubule Incubation System for the Detection of Nephrotoxicants
Authors: Elton Rebecca C. , Rhodes Pauline , Fry Jeffrey R.
Published: April 2019

Many nephrotoxic agents act primarily on proximal tubule cells. Accordingly, the optimal conditions for isolating rat proximal tubule fragments by a collagenase digestion technique and their short-term maintenance have been defined, and the viability of the preparation and its sensitivity to toxicants have been determined. Tubular fragment viability was maintained in incubation for up to 6 hours, as assessed by measuring lactate dehydrogenase leakage, levels of intracellular glutathione, and ATP content. In addition, tubular transport function was maintained, as determined by measuring the uptake of p-aminohippuric acid and α-methyl-glucose, which could be blocked by the selective inhibitors, probenecid and phloridzin, respectively. In this study, the toxicities of allyl alcohol, cephalosporins and cisplatin to proximal tubular fragments were investigated. Allyl alcohol toxicity was greater in tubular fragments isolated from female rats than in those from male rats. Cephaloridine was toxic, while cephalexin and cephalothin were not. These features were consistent with the known in vivo responses to these agents. Toxicity was evident after exposure to cisplatin, with an early reduction in tubular transport being noted. The results highlight the potential of the system described for the isolation and incubation of rat proximal tubule fragments to study xenobiotic-mediated nephrotoxicity. This system could be of benefit in the high-throughput toxicity screening of novel therapeutic agents.
Phytotoxicities of Selected Chemicals and Industrial Effluents to Nitellopsis obtusa Cells, Assessed by Using a Rapid Electrophysiological Charophyte Test
Authors: Manusadžianas Levonas , Vitkus Rimantas , Pörtner Ralf , Märkl Herbert
Published: April 2019

The acute phytotoxicities of seven heavy metals (Cd2+, Cu2+, Hg2+, Ni2+, Zn2+, Cr6+ and Co2+), three phenolic compounds (phenol, 3,5-dichlorophenol and pentachlorophenol) and nine industrial effluents were appraised by using a rapid electrophysiological test with cells of the charophyte, Nitellopsis obtusa. The EC50 values (concentrations causing a 50% decrease in resting potential) obtained for reference chemicals were compared with those of five microbiotests (Polytox®, Microtox®, Selenastrum capricornutum growth inhibition, Daphnia magna immobilisation and Rotoxkit F™) taken from the scientific literature. The 45-minute charophyte test, the freshwater Algaltoxkit F™, Daphtoxkit F™ and Rotoxkit F™ were conducted simultaneously to assess the toxicities of effluents. The Toxkit microbiotests were typically two orders of magnitude more sensitive than the electrophysiological charophyte test to pure chemicals. The electrophysiological charophyte test was generally more sensitive than the Toxkit microbiotests to complex effluents. The rapid electrophysiological test, employing the 45-minute membrane depolarisation of N. obtusa cells as an endpoint, demonstrated similar sensitivity to heavy metals and phenolic compounds as the 20-minute bacterial Polytox® test, but less sensitivity than the 15-minute Microtox® test. Therefore, this rapid macroalgal test appears to be valuable as a sublethal toxicity screening tool for effluents.
Apoptosis versus Cytotoxicity in HeLa Cells Exposed to Paracetamol
Authors: Ruppová Katarína , Urbančíková Miroslava , Wsólová Ladislava , Slameňová Darina
Published: April 2019

Apoptosis is a programmed form of cell death which occurs in response to specific stimuli. It is distinguished from necrotic or accidental cell death by unique events, including the degradation of chromatin and a loss of cellular volume. In contrast to necrotic cell death, cell membrane integrity and mitochondrial function are thought to be maintained until the apoptotic process is well advanced. One of the novel assays for detecting apoptosis is flow cytometry. In our experiments, we used a flow cytometric assay to detect DNA changes in a human cell line (HeLa) exposed to paracetamol, by measuring propidium iodide binding. We were able to detect the apoptotic process in cells exposed to paracetamol. Apoptosis did not correlate with cytotoxicity, and was only found in samples exposed to 4–5mg/ml paracetamol for 8 hours in minimum essential medium and incubated in fresh medium without paracetamol for 14–19 hours. The greatest effect was noted 18 hours after paracetamol exposure. These results were confirmed by studying cell morphology and chromatin condensation by fluorescent microscopy with the fluorochromes acridine orange and ethidium bromide. Our results support the hypothesis that, in cultured cells, apoptosis is induced by a relatively narrow range of chemical concentrations which are known to inhibit the cell cycle, and that apoptosis and inhibition of cell proliferation coincide to some degree.
Retinal Pigment Epithelial Cell Cultures as a Tool for Evaluating Retinal Toxicity In Vitro
Authors: Tähti Hanna , Mäenpää Hanna , Salminen Lotta , Toimela Tarja
Published: April 2019

This article reviews in vitro testing of retinal toxicity in retinal pigment epithelium (RPE) cell cultures. It is based on the literature on RPE cell cultures and on our recent studies on the retinal toxicity of selected amphiphilic drugs. The RPE plays a major role in maintaining the homeostasis and health of the retina. Various pharmacological agents are known to cause adverse effects in RPE cells. For example, long-term treatment with chloroquine in patients with rheumatoid arthritis has induced retinopathy, and tamoxifen, a drug that is commonly used in the treatment of advanced breast cancer and in the prevention of breast cancer among high-risk women, has been reported to cause retinal changes and impaired vision. During our research, we have developed novel in vitro methods for evaluating the retinal toxicity of xenobiotics. We have used a pig RPE primary culture and a human RPE cell line (D407), which retain epithelial cell characteristics. They form a layer of hexagonal cells with intercellular junctions, and possess a keratin-containing cytoskeleton. They are both good models for determining the retinal cell toxicity of test compounds. Further studies on phagocytic activity, lysosomal enzyme activity and glutamate uptake might generate new methods for the toxicological evaluation of the retinal side-effects of drugs in vitro.
Cellular and Molecular Targets of Benzo[a]pyrene and Metal Toxicity in Xenopus laevis Embryos and in Hep G2 Cells
Authors: Camatini Marina , Bonfanti Patrizia , Colombo Anita , Urani Chiara , Crippa Silvia
Published: April 2019

This paper describes the use of two in vitro systems (stage 35 Xenopus laevis embryos and the human hepatoblastoma cell line, Hep G2) to study effects of some environmental contaminants (benzo[a]pyrene, copper and zinc), which are representative of chemicals with different cell targets and mechanisms of action. The ability to activate benzo[a]pyrene and to metabolise it with the cytochrome P4501A isozyme were demonstrated in both in vitro systems by assessing the formation of water-soluble and protein-bound benzo[a]pyrene metabolites and by immunochemical analysis. In X. laevis embryos, the formation of DNA adducts demonstrated the ability to produce benzo[a]pyrene reactive metabolites. Moreover, in Hep G2 cells, the cytoskeletal protein, tubulin, and the reduced form of glutathione proved to be the cellular targets of copper and zinc toxicity. In response to metal-induced stress in Hep G2 cells, there was a cytoplasmic reorganisation of heat shock protein, Hsp 70. In conclusion, the in vitro systems provide for a rapid evaluation of heterogeneous compounds such as benzo[a]pyrene and heavy metals that differ in toxic potency and mechanisms of action. They could also be used to study the mechanisms of toxic action and to identify specific cellular and molecular targets.
Gastrulating Rat Embryo in a Serum-free Culture Model: Changes of Development Caused by Teratogen 5-Azacytidine
Authors: Bulić-Jakuš Floriana , Vlahović Maja , Jurić-Lekić Gordana , Crnek-Kunstelj Vesna , Šerman Draško
Published: April 2019

Developmental processes in gastrulating rat embryos were investigated by using an original, serum-free, chemically defined model system. 9.5-day-old rat embryos, without extraembryonic membranes, were cultivated at the air–liquid interface in a serum-free medium, with and without a protein supplement, for 2 weeks. A teratogenic, demethylating agent, 5-azacytidine, was added to serum-free and protein-free culture medium and to serum-free medium supplemented with human transferrin. A single dose of 5-azacytidine impaired the survival, growth and differentiation of embryos in protein-free medium and serum-free medium with transferrin. In contrast, repeated exposure to 5-azacytidine was required to impair growth in serum-supplemented medium. It was concluded that the activity of 5-azacytidine was easier to detect in a simple, chemically defined medium than in a serum-supplemented medium. This serum-free in vitro method could be useful in screening for teratogenic or embryotoxic substances during gastrulation, the most critical stage of mammalian development.
Description of a Dynamic In Vitro Model of the Dog Gastrointestinal Tract and an Evaluation of Various Transit Times for Protein and Calcium
Authors: Smeets-Peeters Marianne J.E. , Minekus Mans , Havenaar Robert , Schaafsma Gertjan , Verstegen Martin W.A.
Published: April 2019

In order to manufacture complete and balanced dog diets, it is important to know the nutrient requirements of dogs and the availability of these nutrients from food. As pet food manufacturers are restricted in their options for (invasive) animal studies, due to ethical constraints, it is important to have alternative methods for researching the effects of various dog diets. To simulate the gastrointestinal tract of the dog, the dynamic gastrointestinal tract model developed by Minekus et al. was further developed and modified in this study. The model consists of four compartments which simulate the stomach and small intestine (duodenum, jejunum and ileum). Each compartment is made of glass, with a flexible inner wall. This wall can be compressed by increasing the pressure of the surrounding water, mimicking the peristaltic movements and mixing seen in vivo. The model is computer-controlled to simulate physiological parameters such as pH, transit time and secretion of digestive juices, as derived from the literature. Gastric meal delivery and the effects of intestinal transit time on protein digestibility and availability for absorption of calcium from dog food were studied to evaluate the model. The gastric meal delivery of dry dog food was identical to a preset curve, which was based on in vivo data from healthy dogs. The emptying time for canned dog food was somewhat slower than the preset values, probably due to the viscosity of the meal. The differences between the preset values and the measured delivery were not significant. The digestibility of protein and the availability of calcium for absorption increased with a longer transit time. A significant difference was found between medium and slow transit times for the nitrogen content in the ileal delivery effluent and the jejunal dialysates (p < 0.05). The same trend was seen for calcium (not significant). The overall conclusion is that the model is a useful tool for mimicking the gastrointestinal tract of dogs. Parameters such as pH, transit time and enzyme activity can be mimicked and can be kept within a physiological range.
Determination of the Starting dose for Acute Oral Toxicity (LD50) Testing in the up and down Procedure (UDP) from Cytotoxicity Data
Authors: Spielmann Horst , Genschow Elke , Liebsch Manfred , Halle Willi
Published: April 2019

To reduce the number of animals used in acute oral toxicity testing, cytotoxicity data (IC50) can be used to determine the starting dose for in vivo testing by applying the standard regression between IC50 and acute oral LD50 values in the Register of Cytotoxicity (RC). In the RC, the correlation between cytotoxicity, represented by the mean IC50 (IC50x), and the acute oral LD50 of rats and/or mice has been determined for 347 chemicals by applying the linear regression model for log-transformed pairs of IC50 versus oral LD50. The standard regression line of the two toxicity parameters is characterised by an intercept a = 0.625 and regression coefficient b = 0.435, and 252 of 347 chemicals (72.6 %) are located within a dose-range differing by not more than 0.699 (factor FG ≤ log 5) from the standard regression line. In the present study, we have used the RC and its IC50/LD50 regression model to predict the LD50 values from cytotoxicity data for nine chemicals which were tested in an evaluation study of the Up and Down Procedure (UDP). For seven of the nine chemicals, LD50 values (mg/kg) predicted from the RC were in the same dose-range as LD50 values determined in vivo, while the dose-range differed by more than one order of magnitude for the two remaining chemicals. Thus, the prediction of LD50 values from cytotoxicity data was promising in this limited data set. It is proposed that a tiered in vitro/in vivo testing approach will reduce animal use in the UDP method. As the first step, the in vitro cytotoxicity of a new chemical is determined. By applying the RC regression and adapting it to the sensitivity of a specific cell line, the LD50 value (mg/kg) can be predicted from the IC50 value. The predicted LD50 dose is then used as the starting dose in the UDP. In the RC model, the precision of the prediction increases with decreasing toxic potential, and the majority of industrial chemicals (around 90%) are not toxic according to EU classification criteria.
A contemporary course on the introduction to computational fluid dynamics
Authors: Miller Steven A E
Published: April 2019

The University of Florida Department of Mechanical and Aerospace Engineering recently created a new senior technical elective in the field of computational fluid dynamics. The main objectives of the class are learning the process of computational fluid dynamics, skepticism, a course project that uses a popular commercial solver, and a course project that involves programming a simplified computational fluid dynamics code. The course covers introductory material, history, grid generation, numerics, equations of motion, boundary conditions, solvers, turbulence models, visualization, and a number of special topics. Skepticism is enforced throughout the course and forces students to justify the validity of their approach and question numerically generated results. Students in the class undertake a course project to predict a fundamental flow-field and compare predictions with excellent measurements from the open literature. They must also create a simplified computational fluid dynamics code to predict turbulent boundary layer flow. Students have integrated these lessons within student groups across the University of Florida. The emphasis of the course is on skepticism and increasing integration with the curriculum and student group activities. We present the class philosophy for teaching undergraduate computational fluid dynamics and the outcomes of the newly developed course.
Influencing interaction: Does technology increase public participation on community journalism websites?
Authors: Speakman Burton
Published: April 2019

Community media maintain a tradition of interaction, yet, few studies focus on digital operations at smaller publications. This study indicates direct communication matters more than technology to generating interaction advancing interactivity theory. This project further suggests no single technological element likely increases the amount of interaction on the websites of community newspapers and multiple forms of digital outreach could improve the likelihood of success. However, website managers appear indifferent to communicating with the audience online.
Predictors of Home Death in Palliative Care Cancer Patients
Authors: Cantwell Patricia , Turco Sally , Brenneis Carleen , Hanson John , Neumann Catherine M. , Bruera Eduardo
Published: April 2019

With recent changes in health care there is greater emphasis on providing care at home, including the support of families to enable more home deaths. Since a home death may not be practical or desirable in every family situation, there is a need for an objective way to assess the viability of a home death in each individual family situation. The purpose of this study was to describe the relative role of predictors of home death in a cohort of palliative care patients with advanced cancer. A questionnaire was created as a means of assessing the viability of a home death. Five questions were included. Ninety questionnaires were administered by home care coordinators. A follow-up questionnaire was administered to record the place of death. Of the 73 evaluable patients, 34 (47%) died at home and 39 (53%) died in hospital or hospice. The desire for a home death by both the patient and the caregiver, support of a family physician, and presence of more than one caregiver were all significantly associated with a home death. Logistic regression identified a desire for home death by both the patient and the caregiver as the main predictive factor for a home death. The presence of more than one caregiver was also predictive of home death. The questionnaire is simple and, if our results are confirmed, it can be used for predicting those who will not have a home death.
Existential Issues in Palliative Care — Interviews with Cancer Patients
Authors: Bolmsjö Ingrid
Published: April 2019

A minority of terminally ill patients achieve a peaceful death. Many factors, including existential distress, contribute to the emotional disquiet of patients. This study focuses on the reactions of terminally ill cancer patients to questions concerning existential issues within the themes of meaning, relations, autonomy, guilt, dignity, and communication. The results of this study indicate that patients experience a number of problems dealing with existential issues, consider these questions important, and wish to be able to discuss these types of questions with someone.
Frequency of Symptom Distress and Poor Prognostic Indicators in Palliative Cancer Patients Admitted to a Tertiary Palliative Care Unit, Hospices, and Acute Care Hospitals
Authors: Bruera Eduardo , Neumann Catherine , Brenneis Carleen , Quan Hue
Published: April 2019

The Edmonton Regional Palliative Care Program was established to increase access to palliative care for terminal cancer patients in the region. Inpatient care is delivered, in decreasing order of distress, at the tertiary palliative care unit, by consult teams in acute care facilities, and in hospices. We reviewed the admission data for all patients discharged from the program between November 1, 1997, and October 31, 1998, in order to determine if demographical and clinical variables suggested appropriate use of the three levels of care. Patients admitted to the tertiary palliative care unit were significantly younger than those admitted to acute care of hospices (61 years versus 68 years and 71 years respectively, p<0.0001), had a higher frequency of positive screening for alcoholism in the CAGE questionnaire (27% versus 16% and 14% respectively, p<0.0001), and a higher frequency of poor prognostic pain syndromes as compared to acute care admissions (87% versus 65%, p<0.0001). Overall, frequency of symptoms and severe symptoms was significantly higher in patients admitted to the palliative care unit than those admitted to the other two settings. Our results suggest that patients with demographic and clinical indications of higher distress are more frequently admitted to the tertiary palliative care unit. The clinical tools are useful predictors of utilization that can be used for monitoring health care delivery.
Death Education in Home Hospice Care in Japan
Authors: Kawagoe Hiromi , Kawagoe Koh
Published: April 2019

In the practice of home hospice care, death education for both patient and family is extremely important, although little information on its usefulness is available. In this study, the effects of death education under home hospice care were analyzed for 16 patients who died at home. Death education for the patient and his/her family was given at least once in each phase of care, and at least four times in total. The acceptance of death by the patients was judged according to the way they spent their remaining time, to their attitudes, and to their hope for a life after death. Fourteen of 15 patients appeared to accept their own death. An autopsy was performed in five of the 16 cases. In one case, the doctor recommended an autopsy to the family; in the other cases, it was performed in accordance with the patient's or family's wish. As the goal of death education in home hospice care is the acceptance of death by both patient and family, our methods of death education appear to be effective.
Withdrawal of Life Support: How the Family Feels, and Why
Authors: Keenan Sean P. , Mawdsley Cathy , Plotkin Donna , Webster Gregory K. , Priestap Fran
Published: April 2019

The objectives of this study were to develop an instrument to assess the satisfaction of family members with withdrawal of life support (WLS), and to determine which factors are associated with greater levels of satisfaction. To do this, we developed a self-administered questionnaire that was sent to the next-of-kin of intensive care unit (ICU) patients dying following WLS. Over a six-month period, 69 patients died following WLS in the ICU. Three letters were returned “address unknown”, 33 did not respond, and 33 responded, of whom 29 agreed to participate (29/66 = 44% of those contacted). Of these, 24 (83%) strongly agreed with the patient's death being compassionate and dignified, one moderately agreed, one mildly agreed, one was neutral and two strongly disagreed. Items associated with greater satisfaction included: the process of WLS being well explained, WLS proceeding as expected, patient appearing comfortable, family/friends prepared for the decision, appropriate person initiating discussion, adequate privacy during WLS, chance to voice concerns. The study suggests factors that are important to consider in ensuring family comfort with the process of withdrawing life support.