bluuebunny/crossref_metadata_2025 huggingface dataset:
The Sensitivity and Reproducibility of the Zebrafish (Danio rerio) Embryo Test for the Screening of Waste Water Quality and for Testing the Toxicity of Chemicals
Authors: Lahnsteiner Franz
Published: April 2019

The sensitivity of the zebrafish embryo test, a test proposed for routine waste water control, was compared with the acute fish toxicity test, in the determination of six types of waste water and ten different chemicals. The waste water was sampled from the following industrial processes: paper and cardboard production, hide tanning, metal galvanisation, carcass treatment and utilisation, and sewage treatment. The chemicals tested were: dimethylacetamide, dimethylsulphoxide, cadmium chloride, cyclohexane, hydroquinone, mercuric chloride, nickel chloride, nonylphenol, resmethrin and sodium nitrite. For many of the test substances, the zebrafish embryo test and the acute fish toxicity test results showed high correlations. However, there were certain environmentally-relevant substances for which the results of the zebrafish embryo test and the acute fish toxicity test differed significantly, up to 10,000-fold (Hg2+ > 150fold difference; NO2- > 300-fold; Cd2+ > 200-fold; resmethrin > 10,000-fold). For the investigated waste water samples and chemicals, the survival rate of the zebrafish embryos showed high variations between different egg samples, within the range of the EC50 concentration. Subsequently, 5–6 parallel assays were deemed to be the appropriate number necessary for the precise evaluation of the toxicity of the test substances. Also, it was found that the sensitivities of different ontogenetic stages to chemical exposure differed greatly. During the first 12 hours after fertilisation (4-cell stage to the 5-somite stage), the embryos reacted most sensitively to test substance exposure, whereas the later ontogenetic stages showed only slight or no response, indicating that the test is most sensitive during the first 24 hours post-fertilisation.
Comparison of an Automated Pattern Analysis Machine Vision Time-lapse System with Traditional Endpoint Measurements in the Analysis of Cell Growth and Cytotoxicity
Authors: Toimela Tarja , Tähti Hanna , Ylikomi Timo
Published: April 2019

Machine vision is an application of computer vision. It both collects visual information and interprets the images. Although the machine obviously does not ‘see’ in the same sense that humans do, it is possible to acquire visual information and to create programmes to identify relevant image features in an effective and consistent manner. Machine vision is widely applied in industrial automation, but here we describe how we have used it to monitor and interpret data from cell cultures. The machine vision system used (Cell-IQ) consisted of an inbuilt atmosphere-controlled incubator, where cell culture plates were placed during the test. Artificial intelligence (AI) software, which uses machine vision technology, took care of the follow-up analysis of cellular morphological changes. Basic endpoint and staining methods to evaluate the condition of the cells, were conducted in parallel to the machine vision analysis. The results showed that the automated system for pattern analysis of morphological changes yielded comparable results to those obtained by conventional methods. The inbuilt software analysis offers a promising way of evaluating cell growth and various cell phases. The continuous follow-up and label-free analysis, as well as the possibility of measuring multiple parameters simultaneously from the same cell population, were major advantages of this system, as compared to conventional endpoint measurement methodology.
Integrated Decision-tree Testing Strategies for Environmental Toxicity with Respect to the Requirements of the EU REACH Legislation
Authors: Grindon Christina , Combes Robert , Cronin Mark T.D. , Roberts David W. , Garrod John F.
Published: April 2019

Liverpool John Moores University and FRAME recently conducted a research project sponsored by Defra on the status of alternatives to animal testing with regard to the European Union REACH (Registration, Evaluation and Authorisation of Chemicals) system for safety testing and risk assessment of chemicals. The project covered all the main toxicity endpoints associated with the REACH system. This paper focuses on the prospects for using alternative methods (both in vitro and in silico) for environmental (aquatic) toxicity testing. The manuscript reviews tests based on fish cells and cell lines, fish embryos, lower organisms, and the many expert systems and QSARs for aquatic toxicity testing. Ways in which reduction and refinement measures can be used are also discussed, including the Upper Threshold Concentration — Step Down (UTC) approach, which has recently been retrospectively validated by ECVAM and subsequently endorsed by the ECVAM Scientific Advisory Committee (ESAC). It is hoped that the application of this approach could reduce the number of fish used in acute toxicity studies by around 65–70%. Decision-tree style integrated testing strategies are also proposed for acute aquatic toxicity and chronic toxicity (including bioaccumulation), followed by a number of recommendations for the future facilitation of aquatic toxicity testing with respect to environmental risk assessment.
Integrated Decision-tree Testing Strategies for Acute Systemic Toxicity and Toxicokinetics with Respect to the Requirements of the EU REACH Legislation
Authors: Combes Robert , Grindon Christina , Cronin Mark T.D. , Roberts David W. , Garrod John F.
Published: April 2019

Liverpool John Moores University and FRAME conducted a joint research project, sponsored by Defra, on the status of alternatives to animal testing with regard to the European Union REACH (Registration, Evaluation and Authorisation of Chemicals) system for the safety testing and risk assessment of chemicals. The project covered all the main toxicity endpoints associated with REACH. This paper focuses on the use of alternative (non-animal) methods (both in vitro and in silico) for acute systemic toxicity and toxicokinetic testing. The paper reviews in vitro tests based on basal cytotoxicity and target organ toxicity, along with QSAR models and expert systems available for this endpoint. The use of PBPK modelling for the prediction of ADME properties is also discussed. These tests are then incorporated into a decision-tree style, integrated testing strategy, which also includes the use of refined in vivo acute toxicity tests, as a last resort. The implementation of the strategy is intended to minimise the use of animals in the testing of acute systemic toxicity and toxicokinetics, whilst satisfying the scientific and logistical demands of the EU REACH legislation.
Refinement and Optimisation of the Rat CFU-GM Assay to Incorporate the Use of Cryopreserved Bone-marrow Cells for In Vitro Toxicology Applications
Authors: Pessina Augusto , Bonomi Arianna , Baglio Carolina , Cavicchini Loredana , Gribaldo Laura
Published: April 2019

The colony-forming unit-granulocyte–macrophage (CFU-GM) assay has been validated for testing drug haematotoxicity (with both mouse bone-marrow and human cord blood cells) and for predicting in vivo human Maximal Tolerated Dose (MTD) values by extrapolating in vivo data on mouse toxicity. The rat CFU-GM assay is widely used for its capability to evaluate in vitro haematotoxicity, but no standardised procedure suitable for data comparison has been developed. A validated rat CFU-GM assay is needed for many reasons — not least because the rat is the most commonly-used species for the in vivo testing of toxicants. This report describes the refinement and optimisation of a standardised protocol for entering into the prevalidation phase of test development. The sensitivity of rat progenitors to granulocyte–macrophage-colony stimulating factor (GM-CSF), the correlation between the number of cells seeded and the number of colonies obtained, the role of mesenchymal cells on CFU-GM proliferation and the performance of the assay, and the effects of using different types of plastic dishes and sources of cytokines, are described. A standard operating procedure (SOP) based on the use of cryopreserved progenitors has been generated, to be applied to the in vitro toxicity testing of compounds. This SOP dramatically reduces the number of rats used and increases the homogeneity of the data obtained.
The Impact of Introducing Computer-based Alternatives to the Use of Animals in the Teaching of Physiology and Pharmacology at Balkan Universities — A Pilot Study
Authors: Kojic Zvezdana Z. , Dewhurst David G.
Published: April 2019

Balkan universities use a substantial number of small mammals and amphibians in the teaching of physiology and pharmacology. This project investigated whether making computer-based alternatives readily available, and combining this availability with a staff development workshop focusing on methods of integrating such resources into undergraduate curricula, would have any effect on animal use. Teachers from 20 Institutes (from five Balkan countries) participated in the workshop. They presented information about animal use in teaching in their universities, and agreed to introduce at least one computer-based alternative into their teaching in the following year. They were surveyed by questionnaire before, during, and one year after, attending the workshop, in order to estimate any changes in animal use. The results showed a significant ( P < 0.01) reduction in animal use and a high level of implementation of the alternatives provided at the workshop. Teachers recognised the potential benefits of using computers to support their teaching. They lacked knowledge about what computer-based alternatives are available and how to find information about them, including published evidence of their educational effectiveness. In this pilot study, a combination of staff development and making alternatives readily available to teachers had a significant impact on animal use in the teaching of physiology and pharmacology.
An In Vitro Model for Studying Neutrophil Activation During Cardiopulmonary Bypass by Using a Polymerase Chain Reaction Thermocycler
Authors: Tang Min , Zhao Xiao-Gang , Gu Y. John , Chen Chang-Zhi
Published: April 2019

The accurate temperature control of a polymerase chain reaction (PCR) thermocycler was exploited in developing an in vitro model to study neutrophil activation during cardiopulmonary bypass. Neutrophils from 12 volunteers underwent temperature changes in a PCR thermocycler (37°C for 30 minutes, 28°C for 60 minutes, and then 37°C for 90 minutes). Different co-incubates were applied to neutrophils, as follows: Group A: phosphate-buffered saline solution; Group B: platelet activating factor (PAF); Group C: platelet-depleted plasma; Group D: platelet-depleted plasma + PAF; and Group E: platelet-rich plasma. Membrane-bound elastase (MBE) activity was measured every 30 minutes throughout the experiment. MBE acticity decreased significantly after hypothermia, compared with the baseline level ( p < 0.001), and it resumed an increase after re-warming. Among all co-incubates, platelet-rich plasma was the most potent pro-inflammatory stimulus to neutrophils. A linear correlation was found between MBE and platelet count in platelet-rich plasma ( p = 0.004). A novel in vitro model involving a PCR thermocycler has been proved to be reliable in the study of neutrophil activation during cardiopulmonary bypass. The model could possibly be used as an alternative to animals in the development of new drugs to combat neutrophil damage to tissues and organs during cardiopulmonary bypass in cardiac surgery.
The Use of the Zebrafish (Danio rerio) Embryo for the Acute Toxicity Testing of Surfactants, as a Possible Alternative to the Acute Fish Test
Authors: Vaughan Martin , van Egmond Roger
Published: April 2019

At present, the acute toxicity of chemicals to fish is most commonly estimated by means of a short-term test on juvenile or adult animals (OECD TG 203). Although, over the last few years, the numbers used have been reduced due to the implementation of the Three Rs ( Reduction, Refinement and Replacement), significant numbers of fish are still used in acute toxicity tests. With the introduction of the new European Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH) system, this number is likely to increase dramatically. The aim of this work was to test the acute toxicity of a number of anionic, cationic and non-ionic surfactants to embryos of the zebrafish ( Danio rerio), over 48 hours, as a possible alternative to the standard 96-hour fish acute test. We measured the toxicities of 15 surfactants, and compared the results to previously generated adult D. rerio LC50 data (or other fish species, if these data were not available). Comparison of the LC50 data showed that embryos appear to be as sensitive to cationic and non-ionic surfactants as the adult fish, but possibly are more sensitive to anionic surfactants. Toxicity testing with the embryo test can be carried out more quickly than with the adult test, uses much less space and media, requires less effort, and therefore can be performed at a reduced cost. The embryo test may also uncover additional sub-lethal effects, although these were not observed for surfactants. The data presented here show that the 48-hour embryo test can be considered as a suitable alternative to the adult acute fish test for surfactants.
A Comparative Evaluation of In Vitro Skin Sensitisation Tests: The Human Cell-line Activation Test (h-CLAT) versus the Local Lymph Node Assay (LLNA)
Authors: Ashikaga Takao , Sakaguchi Hitoshi , Sono Sakiko , Kosaka Nanae , Ishikawa Makie , Nukada Yuko , Miyazawa Masaaki , Ito Yuichi , Nishiyama Naohiro , Itagaki Hiroshi
Published: April 2019

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.
Introducing Ethical Evaluation of the use of Animals in Experiments in the near East
Authors: Izmirli Serdar , Aldavood S. Javid , Yasar Askin , Phillips Clive J.C.
Published: April 2019

Turkey and Iran are two Near East countries that are developing ethical regulation systems for animal experiments, following significant developments in this field in neighbouring Europe. In Turkey, legislation was passed between 2004 and 2006 to mandate ethical review of experiments which involve the use of laboratory animals. Today, one central and 73 local ethics committees exist, to evaluate the use of animals and the numbers of animals used, and to consider applications for animal experimentation. In Iran, Animal Ethics Committees (AECs) were established in 2004 in approximately 50 medical universities. The objective of the AECs is to preserve animal welfare and ethics, taking into account Islamic principles, and to be cognisant of world trends for the improvement of laboratory animal welfare. The recent establishment of animal ethical review in the Near East could benefit from experiences in Western countries, in their evolution toward high standards of experimental animal ethics.
The Bovine Corneal Opacity and Permeability Test in Routine Ocular Irritation Testing and its Improvement within the Limits of OECD Test Guideline 437
Authors: Schrage Arnhild , Kolle Susanne N. , Moreno Maria C. Rey , Norman Kimberly , Raabe Hans , Curren Rodger , van Ravenzwaay Bennard , Landsiedel Robert
Published: April 2019

Data on eye irritation are generally needed for the hazard identification of chemicals. As the Bovine Corneal Opacity and Permeability (BCOP) test has been accepted by many regulatory agencies for the identification of corrosive and severe ocular irritants since September 2009 (OECD Test Guideline 437, TG 437), we evaluated this alternative method for routine testing at BASF. We demonstrated our technical proficiency by testing the reference standards recommended in TG 437, and 21 additional materials with published BCOP and in vivo data. Our results matched the published in vitro data very well, but with some intentionally selected false negatives (FNs) and false positives (FPs), the concordance was 77% (24/31), with FN and FP rates of 20% (2/10) and 24% (5/21), respectively. In addition, we tested 21 in-house materials, demonstrating the utility of the BCOP assay for our own test material panel. Histopathological assessment of the corneas by light microscopy was also conducted, as this was suggested as a means of improving the identification of FNs. The histopathology corrected the classification of some FNs, but also increased the number of FPs. Parallel to the test method evaluation, we compared three new opacitometer models with the current standard device. We recommend the use of an opacitometer developed in our BASF laboratory, which has certified components and electronic data storage, resulting in what we consider to be excellent sensitivity, stability and reproducibility.
“How Sick Must Your Mouse Be?” — An Analysis of the Use of Animal Models in Huntington's Disease Research
Authors: Franco Nuno H. , Olsson I. Anna S.
Published: April 2019

Refinement measures to improve animal welfare can ease the ethical dilemma between human benefit and animal harm in research with animal models of neurodegenerative diseases. To evaluate the potential for refinement, as well as its implementation in research, we analysed papers on murine models of Huntington's disease (HD) published between 1999 and 2009 ( n = 233). Each study was classified according to a four-level severity scale, in terms of the disease stage that animals were allowed to reach, the execution of invasive procedures, and the implementation of refinement. Reports of ethical approval, and regulatory compliance in general, followed a clear rising trend over the years reviewed ( p < 0.001). However, the proportion of high-severity studies did not change over that period. Also, of the studies for which approval was reported ( n = 120), 36% were attributed the highest severity level. The observed discrepancy between the rising motivation to affirm regulatory compliance, and the unaltered proportion of studies allowing animals to reach severely distressful stages, raises both ethical and methodological issues, which we discuss in this paper.
Experience of the Use of Table-top Simulators as Alternatives in the Primary Surgical Training of Veterinary Undergraduate Students
Authors: Perez-Rivero Juan Jose , Rendon-Franco Emilio
Published: April 2019

Table-top simulators are valuable alternatives to animal models in health sciences training, by virtue of their low cost, ease of construction and ability to successfully fulfil teaching objectives
Animal-free Toxicology: The Use of Human Tissue to Replace the Use of Animals — Examples from Human Biomonitoring and Human Placental Transport Studies
Authors: Knudsen Lisbeth E.
Published: April 2019

Human data on exposure and adverse effects are the most appropriate for human risk assessment, and modern toxicology focuses on human pathway analysis and the development of human biomarkers. Human biomonitoring and human placental transport studies provide necessary information for human risk assessment, in accordance with the legislation on chemical, medicine and food safety. Toxicology studies based on human mechanistic and exposure information can replace animal studies. These animal-free approaches can be further supplemented by new in silico methods and chemical structure–activity relationships. The inclusion of replacement expertise in the international Three Rs centres, the ongoing exploration of alternatives to animal research, and the improvement of conditions for research animals, all imply the beginning of a paradigm shift in toxicology research toward the use of human data.
FRAME's Alternatives Timeline Project
Authors: Coll Kevin
Published: April 2019

FRAME's proposed online resource on alternatives will be easy to access, visually appealing and content rich
Why Animal Studies are Still Being Used in Drug Development
Authors: Kooijman Marlous
Published: April 2019

The lack of success of the implementation of new methods in drug development can be studied from an innovation perspective
Prediction of Acute Aquatic Toxicity toward Daphnia Magna by using the GA-kNN Method
Authors: Cassotti Matteo , Ballabio Davide , Consonni Viviana , Mauri Andrea , Tetko Igor V. , Todeschini Roberto
Published: April 2019

In this study, a QSAR model was developed from a data set consisting of 546 organic molecules, to predict acute aquatic toxicity toward Daphnia magna. A modified k-Nearest Neighbour ( kNN) strategy was used as the regression method, which provided prediction only for those molecules with an average distance from the k nearest neighbours lower than a selected threshold. The final model showed good performance (R2 and Q2cv equal to 0.78, Q2ext equal to 0.72). It comprised eight molecular descriptors that encoded information about lipophilicity, the formation of H-bonds, polar surface area, polaris-ability, nucleophilicity and electrophilicity.
Openness on Animal Research: The Gauntlet has been Thrown Down
Authors: Balls Michael
Published: April 2019

The delivery of the UK Government's and Concordat's commitments to greater openness on animal research is eagerly awaited; meanwhile, the questions raised by two studies on the use of animal tests to predict the toxic effects of drugs in humans should be answered
Lung Fibrosis: Drug Screening and Disease Biomarker Identification with a Lung Slice Culture Model and Subtracted cDNA Library
Authors: Guo Tong , Lok Ka Yee , Yu Changhe , Li Zhuo
Published: April 2019

Pulmonary fibrosis is a progressive and irreversible disorder with no appropriate cure. A practical and effective experimental model that recapitulates the disease will greatly benefit the research community and, ultimately, patients. In this study, we tested the lung slice culture (LSC) system for its potential use in drug screening and disease biomarker identification. Fibrosis was induced by treating rat lung slices with 1ng/ml TGF-ß1 and 2.5μM CdCl2, quantified by measuring the content of hydroxyproline, and confirmed by detecting the expression of collagen type III alpha 1 ( Col3α1) and connective tissue growth factor ( CTGF) genes. The anti-fibrotic effects of pirfenidone, spironolactone and eplerenone were assessed by their capability to reduce hydroxyproline content. A subtractive hybridisation technique was used to create two cDNA libraries (subtracted and unsubtracted) from lung slices. The housekeeping gene glyceraldehyde-3-phosphate dehydrogenase ( GAPDH) was employed to assess the subtraction efficiency of the subtracted cDNA library. Clones from the two libraries were sequenced and the genes were identified by performing a BLAST search on the NCBI GenBank database. Furthermore, the relevance of the genes to fibrosis formation was verified. The results presented here show that fibrosis was effectively induced in cultured lung slices, which exhibited significantly elevated levels of hydroxyproline and Col3α1/CTGF gene expression. Several inhibitors have demonstrated their anti-fibrotic effects by significantly reducing hydroxyproline content. The subtracted cDNA library, which was enriched for differentially expressed genes, was used to successfully identify genes associated with fibrosis. Collectively, the results indicate that our LSC system is an effective model for the screening of drug candidates and for disease biomarker identification.
12. William Russell: Polymath, Wordsmith, Classicist and Humourist
Authors: Balls Michael
Published: April 2019

Reading the The Principles of Humane Experimental Technique is a very special privilege, not only because of the importance of the messages it conveys, but because of the unique character and qualities of its principal author
Human-based Systems in Drug and Chemical Safety Testing — toward Replacement, the ‘Single R’
Authors: Coleman Robert A.
Published: April 2019

The Three Rs was a concept originally conceived as a means of reducing the suffering of laboratory animals that are used largely in identifying any potential safety issues with chemicals to which humans may be exposed. However, with growing evidence of the shortcomings of laboratory animal testing to reliably predict human responsiveness to such chemicals, questions are now being asked as to whether it is appropriate to use animals as human surrogates at all. This raises the question of whether, of the original Three Rs, two — Reduction and Refinement — are potentially redundant, and whether, instead, we should concentrate on the third R: Replacement. And if this is the best way forward, it is inevitable that this R should be based firmly on human biology. The present review outlines the current state-of-the-art regarding our access to human biology through in vitro, in silico and in vivo technologies, identifying strengths, weaknesses and opportunities, and goes on to address the prospect of achieving a single R, with some suggestions as to how to progress toward this goal.
2014 Lush Science Prize
Authors: 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. 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. This Background Paper includes information on recent work by the relevant scientific institutions and projects in this area, including AXLR8, OECD, CAAT, 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, with a view to presenting the two papers receiving the highest score to the judges for consideration.
Humane Education in Brazil: Organisation, Challenges and Opportunities
Authors: Bachinski Róber , Tréz Thales , Alves Gutemberg G. , Garcia Rita de C.M. , Oliveira Simone T. , Alonso Luciano da S. , Heck Júlio X. , Dias Claudia M.C. , Neto João M. Costa , Rocha Alexandro A. , Ruiz Valeska R.R. , Paixão Rita L.
Published: April 2019

Humane education and the debate on alternatives to harmful animal use for training is a relatively recent issue in Brazil. While animal use in secondary education has been illegal since the late 1970s, animal use in higher science education is widespread. However, alternatives to animal experiments in research and testing have recently received attention from the Government, especially after the first legislation on animal experiments was passed, in 2008. This article proposes that higher science education should be based on a critical and humane approach. It outlines the recent establishment of the Brazilian Network for Humane Education (RedEH), as a result of the project, Mapping Animal Use for Undergraduate Education in Brazil, which was recognised by the 2014 Lush Prize. The network aims to create a platform to promote change in science education in Brazil, starting by quantitatively and qualitatively understanding animal use, developing new approaches adapted to the current needs in Brazil and Latin America, and communicating these initiatives nationally. This paper explores the trajectory of alternatives and replacement methods to harmful animal use in training and education, as well as the status of humane education in Brazil, from the point of view of educators and researchers engaged with the network.
A New Approach to Optimising the Use of Animal Models in Drug Discovery through Big Data Sharing
Authors: Lu Jiaqi , Wang Jianfei
Published: April 2019

Sharing full details of animal models would increase the consistency of models between establishments and reduce the numbers of animals used for set-up and validation
19. Russell and Burch after 1959
Authors: Balls Michael
Published: April 2019

After the publication of The Principles, Russell and Burch didn't meet again for more than 30 years; however, when they were rediscovered, and rediscovered each other, they had a rewarding few years, during which many others benefited from their wisdom and their insight
Comparing the CORAL and Random Forest Approaches for Modelling the In Vitro Cytotoxicity of Silica Nanomaterials
Authors: Cassano Antonio , Robinson Richard L. Marchese , Palczewska Anna , Puzyn Tomasz , Gajewicz Agnieszka , Tran Lang , Manganelli Serena , Cronin Mark T.D.
Published: April 2019

Nanotechnology is one of the most important technological developments of the 21st century. In silico methods to predict toxicity, such as quantitative structure–activity relationships (QSARs), promote the safe-by-design approach for the development of new materials, including nanomaterials. In this study, a set of cytotoxicity experimental data corresponding to 19 data points for silica nanomaterials were investigated, to compare the widely employed CORAL and Random Forest approaches in terms of their usefulness for developing so-called ‘nano-QSAR’ models. ‘External’ leave-one-out cross-validation (LOO) analysis was performed, to validate the two different approaches. An analysis of variable importance measures and signed feature contributions for both algorithms was undertaken, in order to interpret the models developed. CORAL showed a more pronounced difference between the average coefficient of determination (R2) for training and for LOO (0.83 and 0.65 for training and LOO, respectively), compared to Random Forest (0.87 and 0.78 without bootstrap sampling, 0.90 and 0.78 with bootstrap sampling), which may be due to overfitting. With regard to the physicochemical properties of the nanomaterials, the aspect ratio and zeta potential were found to be the two most important variables for Random Forest, and the average feature contributions calculated for the corresponding descriptors were consistent with the clear trends observed in the data set: less negative zeta potential values and lower aspect ratio values were associated with higher cytotoxicity. In contrast, CORAL failed to capture these trends.
The Novel Induction of Retinal Ganglion Cell Apoptosis in Porcine Organ Culture by NMDA — An Opportunity for the Replacement of Animals in Experiments
Authors: Kuehn Sandra , Hurst Jose , Jashari Adelina , Ahrens Kathrin , Tsai Teresa , Wunderlich Ilan M. , Dick H. Burkhard , Joachim Stephanie C. , Schnichels Sven
Published: April 2019

Some of the advantages of retina organ culture models include their efficient and easy handling and the ability to standardise relevant parameters. Additionally, when porcine eyes are obtained from the food industry, no animals are killed solely for research purposes. To induce retinal degeneration, a commonly used toxic substance, N-methyl-D-aspartate (NMDA), was applied to the cultures. To this end, organotypic cultures of porcine retinas were cultured and treated with different doses of NMDA (0 [control], 50, 100 and 200μM) on day 2 for 48 hours. On day 7, the retinas were cryo-conserved for histological, Western blot and quantitative rt-PCR (qrt-PCR) analyses. NMDA treatment was found to significantly increase retinal ganglion cell (RGC) apoptosis in all the treated groups, without a profound RGC loss. In addition, the intrinsic apoptotic pathway was activated in the 50μM and 100μM NMDA groups, whereas induced nitric oxide synthase ( iNOS) expression was increased in the 200μM group. A slight microglial response was detectable, especially in the 100μM group. NMDA treatment induced apoptosis, oxidative stress and a slight microglia activation. All these effects mimic a chronic slow progressive disease that especially affects RGCs, such as glaucoma. A particular advantage of this model is that mediators that can interact in the very early stages of the onset of RGC death, can be easily detected and potential therapies can be tested.
The 2017 Lush Prize Awards
Authors: BéruBé Kelly , Redmond Craig
Published: April 2019

This year saw eighteen prizewinning projects from eleven countries, and a new award in recognition of the work of Andrew Tyler
Comparative Surfactant Reactivity of Canine and Human Stratum Corneum: A Plea for the Use of the Corneosurfametry Bioassay
Authors: Goffin Véronique , Fontaine Jacques , Piérard Gérald E.
Published: April 2019

Comparative dermatology has paid little attention to the physiopathology of the stratum corneum. In this study, we investigated the responses of human and canine horny layers to marketed animal wash products by using the corneosurfametry bioassay. Previous work has shown that, with increasing surfactant aggressiveness to the stratum corneum, the colorimetric index of mildness (CIM) decreases, while both the corneosurfametry index (CSMI) and the overall difference in corneosurfametry (ODC) increase. In the present study, stratum corneum reactivity to wash products and inter-individual variability were significantly higher in humans than in dogs. For the three corneosurfametry variables, linear correlations were found between data gathered in the two panel groups. In conclusion, this pilot study suggests that mean stratum corneum reactivity to surfactants is stronger in humans than in dogs. Inter-individual variation, indicative of sensitive skin, also appears to be broader in humans. As a consequence, data gathered from dogs by using the corneosurfametry bioassay cannot be extrapolated to humans. Such variation between species could be important in the assessment of product safety and in supporting claims for mildness.
Short-term Toxicity of Various Pharmacological Agents on the In Vitro Nitrification Process in a Simple Closed Aquatic System
Authors: Nimenya Herman , Delaunois Annie , La Duong Duc , Bloden Serge , Defour Jean , Nicks Baudouin , Ansay Michel
Published: April 2019

During the treatment of fish diseases, drugs which inhibit the nitrification process can cause acute ammonia toxicity. The same phenomenon can occur when fish are put into a tank without active cultures of nitrifying bacteria. The purpose of this study was to quantify the inhibitory effects of 15 pharmacological agents, which are often used as therapeutic agents in ichthyopathology, on ammonia removal and nitrate production in a simple closed aquatic system. The experiments were conducted in polyethylene bags containing activated biofilters and synthetic water solutions, held in a water bath. Ammonia was added to initiate the nitrification process, and graded concentrations of various pharmacological agents were added. The effects of the pharmacological agents on in vitro nitrification were assessed by monitoring ammonia and nitrate concentrations compared to controls with no added agents, for 24 hours. Graded concentrations of ampicillin (Albipen®), chloramine T, enrofloxacin (Baytril®), erythromycin, levamisole, methylene blue and polymyxin B induced dose-dependent inhibitions of ammonia removal and nitrate production. The corresponding linear regression curves showed high correlation coefficients and were highly significant (p < 0.05). The addition of chloramphenicol, copper (II) sulphate, kanamycin disulphate, malachite green, neomycin sulphate, potassium penicillin G, tetracycline and a mixture of trimethoprim and sulphadoxin (Duoprim™) had no significant effects on the nitrification process. A significant dose-related inhibition of nitrate production, but not of ammonia oxidation, was observed with enrofloxacin. The significant correlation (r = 0.940; p < 0.001) between the degrees of inhibition of ammonia oxidation and nitrate production for the various inhibitory pharmacological agents has also been calculated, with a view to validating this method. The data presented suggest that separate tank facilities for hospitalisation or quarantine are necessary when treating diseased fish with ampicillin, enrofloxacin, chloramine T, erythromycin, levamisole, methylene blue or polymyxin B, in order to avoid ammonia poisoning.
Effect of ex vivo extracorporeal membrane oxygenation flow dynamics on immune response
Authors: Ki Katrina K , Passmore Margaret R , Chan Chris Hoi Houng , Malfertheiner Maximillian V , Bouquet Mahe , Cho Hwa Jin , Suen Jacky Y , Fraser John F
Published: April 2019

Background:

Extracorporeal membrane oxygenation is a life-saving support for heart and/or lung failure patients. Despite technological advancement, abnormal physiology persists and has been associated with subsequent adverse events. These include thrombosis, bleeding, systemic inflammatory response syndrome and infection. However, the underlying mechanisms are yet to be elucidated. We aimed to investigate whether the different flow dynamics of extracorporeal membrane oxygenation would alter immune responses, specifically the overall inflammatory response, leukocyte numbers and activation/adhesion surface antigen expression.

Methods:

An ex vivo model was used with human whole blood circulating at 37°C for 6 hours at high (4 L/minute) or low (1.5 L/minute) flow dynamics, with serial blood samples taken for analysis.

Results:

During high flow, production of interleukin-1β (p < 0.0001), interleukin-6 (p = 0.0075), tumour necrosis factor-α (p = 0.0013), myeloperoxidase (p < 0.0001) and neutrophil elastase (p < 0.0001) were significantly elevated over time compared to low flow, in particular at 6 hours. While the remaining assessments exhibited minute changes between flow dynamics, a consistent trend of modulation in leukocyte subset numbers and phenotype was observed at 6 hours.

Conclusion:

We conclude that prolonged circulation at high flow triggers a prominent pro-inflammatory cytokine response and activates neutrophil granule release, but further research is needed to better characterize the effect of flow during extracorporeal membrane oxygenation.
Assessment of Emission Reduction Scenarios with a Focus on the Impact of Vehicle Fleets on Tehran Air Quality: Case Study
Authors: Shahbazi Hossein , Hosseini Vahid , Torbatian Sara , Hamedi Masoud
Published: April 2019

In recent years, the city of Tehran has frequently experienced severe air pollution episodes, and the increase in the use of vehicles has exacerbated this critical situation. It is unclear to what extent each vehicle category contributes to the production of pollution during a pollution event. For this purpose, a combination of the weather research and forecasting (WRF) and the Comprehensive Air Quality Model with Extensions (CAMx) modeling systems was used to simulate the distributions of pollutant concentrations over Tehran in a pollution episode from December 3 to 23, 2017. First, the performance of the modeling system was evaluated with respect to the simulation outputs against observations. Results showed satisfactory performance by the models in predicting meteorological parameters and pollutant concentrations. There were some discrepancies between measured and simulated pollutant levels that might have several causes, including, for example, not considering dust emissions in the system. The second step focused on the assessment of different emission scenarios to quantify the impact of each vehicle fleet in producing pollution during such severe episodes. For this reason, eight different scenarios were considered in which the emissions of each vehicle category were entirely eliminated from the corresponding scenario. The results show that the removal of passenger cars and motorcycles could have the most significant impact on PM2.5 reduction, with a maximum drop of 32% and 30%, respectively. In addition, the NO2 concentrations could be reduced by about 20% and 10% as a result of the removal of passenger cars and motorcycles, respectively. Finally, two traffic schemes with a main focus on the role of motorcycles on the degradation of Tehran’s air quality were also examined for the same episode.
An electrospun citric acid modified polyvinyl alcohol scaffold for vascular tissue engineering
Authors: Velutheril Thomas Lynda , Nair Prabha Damodaran
Published: April 2019

The main aim of this study is to fabricate an electrospun citric acid modified polyvinyl alcohol polyester that is biodegradable with non-toxic by-products and can be used for the culture of vascular smooth muscle cells. In this study, we have optimized the conditions for the electrospinning process of this polyester. The fibre morphology was studied by scanning electron microscopy which indicated that the fibre diameter was optimum at a range of 200 to 700 µm at 5% concentration and flow rate of 0.3 mL/h. The membranes were characterized for the change in structural aspects at the molecular level. The results showed development of more crystalline domains on electrospinning. The surface characteristics were also explored. Cell culture studies confirmed that the electrospun scaffold supported the attachment and proliferation of smooth muscle cells, which was evident from the cell proliferation assay. Hence, the electrospun polyester scaffolds are non-toxic and biocompatible with vascular smooth muscle cells, and find promising potential as scaffolds for vascular tissue engineering.
Reconfiguration control method for non-redundant actuator faults on unmanned aerial vehicle
Authors: Boche Adèle , Farges Jean-Loup , De Plinval Henry
Published: April 2019

In this article, an indirect continuous and discrete Fault-Tolerant Control scheme is applied to the longitudinal dynamics of a fixed-wing unmanned aerial vehicle. To address the problem raised by a possible engine fault, the FTC method is adapted to non-redundant actuator faults by considering signatures on residuals, designing discounted cost optimal controllers, using rolling horizon techniques and considering actuator dynamics in the state equation. The efficiency of the proposed approach is demonstrated on a scenario involving thrust and elevon faults during a landing procedure.
Nondihydropyridine Calcium Channel Blockers for the Treatment of Proteinuria: A Review of the Literature
Authors: Steuber Taylor D. , Lee Jenna , Holloway Amanda , Andrus Miranda R.
Published: April 2019

Objective: To review the use of nondihydropyridine calcium channel blockers (non-DHP CCBs) for the treatment of proteinuria in diabetic and nondiabetic kidney disease. Data Sources: A search using PubMed and MEDLINE, Scopus, and Google Scholar was performed from 1964 through February 2019 using the following search terms alone or in combination: verapamil, diltiazem, non-dihydropyridine calcium channel blocker, proteinuria, albuminuria, microalbuminuria, kidney disease, renal disease. Study Selection and Data Extraction: All prospective English-language trials examining one or more non-DHP CCB for the treatment of proteinuria were evaluated. Data Synthesis: A total of 13 clinical trials examining the use of non-DHP CCBs to treat proteinuria alone or in combination with angiotensin-converting enzyme (ACE) inhibitors or angiotensin receptor blockers (ARBs) were included in the evaluation. Most studies evaluated patients with macroalbuminuria secondary to diabetes and hypertension. Verapamil was the most common agent studied. Non-DHP CCBs were effective in reducing proteinuria in diabetic kidney disease but did not reduce renal or cardiovascular outcomes in the one trial that evaluated clinical end points. They were generally well tolerated, with the most common adverse effect reported being constipation. Relevance to Patient Care and Clinical Practice: This review evaluates and summarizes the available evidence on non-DHP CCBs for treatment of proteinuria in patients with existing kidney disease. Conclusion: Non-DHP CCBs may be a reasonable therapeutic option for patients with diabetic kidney disease and persistent proteinuria despite maximum doses of ACE inhibitors or ARBs. Additionally, they may be reasonable alternatives to ACE inhibitors or ARBs if a contraindication or intolerance exists.
Surgical Tips for Performing Open Fasciotomies for Chronic Exertional Compartment Syndrome of the Leg
Authors: Scully William F. , Benavides Jerome M.
Published: April 2019

Chronic exertional compartment syndrome (CECS) of the leg is a debilitating condition that has previously been characterized in athletes and military personnel. The results of surgical management in these patient populations have previously been published with inconsistencies in surgical methods and patient outcomes noted. While endoscopic and minimally invasive techniques have been described, a detailed description of a “standard” open, 2-incision lateral and single-incision medial technique for the treatment of CECS has not been previously published. The purpose of this technique article is to highlight several tips and considerations related to this procedure in the hopes of standardizing treatment and potentially improving patient outcomes.

Level of Evidence: Level V, expert opinion.
The Measurement of Green Workplace Behaviors: A Systematic Review
Authors: Francoeur Virginie , Paillé Pascal , Yuriev Alexander , Boiral Olivier
Published: April 2019

The literature on the green behaviors adopted by individuals in workplace settings has grown significantly over the past decade. Many studies have examined the factors associated with individual actions. However, the comparability of the studies conducted on the subject is a common concern, mainly because of the wide range of measurement tools based on different sets of items used in such research. Therefore, the aim of this study is to determine the degree of methodological maturity of green workplace behaviors based on a systematic review of research published on the subject between 1977 and 2016. Five major trends were identified from the 53 papers reviewed as part of this research: (a) the predominance of scales for measuring “green office” behaviors, (b) the redundancy of certain items, (c) the limited efforts devoted to measuring counterproductive green behaviors, (d) the emergence of new subcategories of proenvironmental behaviors, and (e) and the abundance of scales measuring voluntary green behaviors (extra-role). Through an analysis of existing measurement tools, this article proposes a decision tree designed to help scholars choose appropriate items for their studies. This may, in turn, contribute to the literature on green workplace behaviors by reducing bias and limiting the unnecessary creation of new measurement scales.