Advancing the 3Rs: innovation, implementation, ethics and society

The 3Rs principle of replacing, reducing and refining the use of animals in science has been gaining widespread support in the international research community and appears in transnational legislation such as the European Directive 2010/63/EU, a number of national legislative frameworks like in Switzerland and the UK, and other rules and guidance in place in countries around the world. At the same time, progress in technical and biomedical research, along with the changing status of animals in many societies, challenges the view of the 3Rs principle as a sufficient and effective approach to the moral challenges set by animal use in research. Given this growing awareness of our moral responsibilities to animals, the aim of this paper is to address the question: Can the 3Rs, as a policy instrument for science and research, still guide the morally acceptable use of animals for scientific purposes, and if so, how? The fact that the increased availability of alternatives to animal models has not correlated inversely with a decrease in the number of animals used in research has led to public and political calls for more radical action. However, a focus on the simple measure of total animal numbers distracts from the need for a more nuanced understanding of how the 3Rs principle can have a genuine influence as a guiding instrument in research and testing. Hence, we focus on three core dimensions of the 3Rs in contemporary research: (1) What scientific innovations are needed to advance the goals of the 3Rs? (2) What can be done to facilitate the implementation of existing and new 3R methods? (3) Do the 3Rs still offer an adequate ethical framework given the increasing social awareness of animal needs and human moral responsibilities? By answering these questions, we will identify core perspectives in the debate over the advancement of the 3Rs

The FLUXNET2015 dataset and the ONEFlux processing pipeline for eddy covariance data

The FLUXNET2015 dataset provides ecosystem-scale data on CO2, water, and energy exchange between the biosphere and the atmosphere, and other meteorological and biological measurements, from 212 sites around the globe (over 1500 site-years, up to and including year 2014). These sites, independently managed and operated, voluntarily contributed their data to create global datasets. Data were quality controlled and processed using uniform methods, to improve consistency and intercomparability across sites. The dataset is already being used in a number of applications, including ecophysiology studies, remote sensing studies, and development of ecosystem and Earth system models. FLUXNET2015 includes derived-data products, such as gap-filled time series, ecosystem respiration and photosynthetic uptake estimates, estimation of uncertainties, and metadata about the measurements, presented for the first time in this paper. In addition, 206 of these sites are for the first time distributed under a Creative Commons (CC-BY 4.0) license. This paper details this enhanced dataset and the processing methods, now made available as open-source codes, making the dataset more accessible, transparent, and reproducible.Peer reviewe

Effekten av navigerbarhet på e-handel för studenter

The consumer trend towards sustainable purchasing habits online has increased due to the environmental impact of the clothing industry. While second-hand shopping is a popular option, it is often avoided due to the perceived inconvenience. The purpose of this study was to provide a basis for creating a navigable website for students in the context of second hand shopping. The research was conducted through a combination of surveys, user testing, and data analysis. Perceived navigability was measured using CTA protocol, CSUQ, and a Lostness calculation. Using standardised theoretical design protocols, a high rate of navigability was perceived for the first iteration of the website. By applying the feedback from the user test, adequate changes such as improved error messages and improved structural navigation could be made which improved the navigability further. This study underscores the importance of accessibility and customisation tailored to the intended customers' requisites. The main conclusion made in this study was that even though standard design protocols are followed, minor details such as buttons that are difficult to click or vague error messages, can significantly hinder navigation.Konsumenttrenden mot hållbara inköpsvanor online har ökat på grund av klädindustrins miljöpåverkan. Trots att andrahandsmarknaden på nätet är ett populärt alternativ för vissa konsumenter, undviks den ofta på grund av en alltför besvärlig upplevelse. Syftet med denna studie var att ge en grund för att skapa en navigerbar webbplats för studenter i sammanhanget av second hand-shopping. Forskningen utfördes genom en kombination av enkäter, användartester och dataanalys. Uppfattad navigerbarhet mättes med hjälp av CTA-protokoll, CSUQ och en "Lostness" -beräkning. Med hjälp av standardiserade designteorier kunde en hög navigerbarhet upplevas i den första iterationen av webbplatsen. Genom att tillämpa feedback från användartesten kunde lämpliga ändringar så som förbättrade felmeddelanden och förbättrad navigeringsstrukturgenomföras som ytterligare förbättrade navigerbarheten. Resultaten i denna studie understryker vikten av tillgänglighet och anpassning vilken är skräddarsydd efter avsedda kundernas krav. Den huvudsakliga slutsatsen dragen i studien var att även om standardiserade designprotokoll följs kan mindre detaljer som svårtryckta knappar och vaga felmeddelanden markant hindra navigeringen

Effekten av navigerbarhet på e-handel för studenter

The consumer trend towards sustainable purchasing habits online has increased due to the environmental impact of the clothing industry. While second-hand shopping is a popular option, it is often avoided due to the perceived inconvenience. The purpose of this study was to provide a basis for creating a navigable website for students in the context of second hand shopping. The research was conducted through a combination of surveys, user testing, and data analysis. Perceived navigability was measured using CTA protocol, CSUQ, and a Lostness calculation. Using standardised theoretical design protocols, a high rate of navigability was perceived for the first iteration of the website. By applying the feedback from the user test, adequate changes such as improved error messages and improved structural navigation could be made which improved the navigability further. This study underscores the importance of accessibility and customisation tailored to the intended customers' requisites. The main conclusion made in this study was that even though standard design protocols are followed, minor details such as buttons that are difficult to click or vague error messages, can significantly hinder navigation.Konsumenttrenden mot hållbara inköpsvanor online har ökat på grund av klädindustrins miljöpåverkan. Trots att andrahandsmarknaden på nätet är ett populärt alternativ för vissa konsumenter, undviks den ofta på grund av en alltför besvärlig upplevelse. Syftet med denna studie var att ge en grund för att skapa en navigerbar webbplats för studenter i sammanhanget av second hand-shopping. Forskningen utfördes genom en kombination av enkäter, användartester och dataanalys. Uppfattad navigerbarhet mättes med hjälp av CTA-protokoll, CSUQ och en "Lostness" -beräkning. Med hjälp av standardiserade designteorier kunde en hög navigerbarhet upplevas i den första iterationen av webbplatsen. Genom att tillämpa feedback från användartesten kunde lämpliga ändringar så som förbättrade felmeddelanden och förbättrad navigeringsstrukturgenomföras som ytterligare förbättrade navigerbarheten. Resultaten i denna studie understryker vikten av tillgänglighet och anpassning vilken är skräddarsydd efter avsedda kundernas krav. Den huvudsakliga slutsatsen dragen i studien var att även om standardiserade designprotokoll följs kan mindre detaljer som svårtryckta knappar och vaga felmeddelanden markant hindra navigeringen

A novel variant of FGFR3 causes proportionate short stature

Objective: Mutations of the fibroblast growth factor receptor 3 (FGFR3) cause various forms of short stature, of which the least severe phenotype is hypochondroplasia, mainly characterized by disproportionate short stature. Testing for an FGFR3 mutation is currently not part of routine diagnostic testing in children with short stature without disproportion. Design: A three-generation family A with dominantly transmitted proportionate short stature was studied by whole-exome sequencing to identify the causal gene mutation. Functional studies and protein modeling studies were performed to confirm the pathogenicity of the mutation found in FGFR3. We performed Sanger sequencing in a second family B with dominant proportionate short stature and identified a rare variant in FGFR3. Methods: Exome sequencing and/or Sanger sequencing was performed, followed by functional studies using transfection of the mutant FGFR3 into cultured cells; homology modeling was used to construct a three-dimensional model of the two FGFR3 variants. Results: A novel p.M528I mutation in FGFR3 was detected in family A, which segregates with short stature and proved to be activating in vitro. In family B, a rare variant (p.F384L) was found in FGFR3, which did not segregate with short stature and showed normal functionality in vitro compared with WT. Conclusions: Proportionate short stature can be caused by a mutation in FGFR3. Sequencing of this gene can be considered in patients with short stature, especially when there is an autosomal dominant pattern of inheritance. However, functional studies and segregation studies should be performed before concluding that a variant is pathogenic

Global, in situ analysis of the structural proteome in individuals with Parkinson's disease to identify a new class of biomarker

Parkinson's disease (PD) is a prevalent neurodegenerative disease for which robust biomarkers are needed. Because protein structure reflects function, we tested whether global, in situ analysis of protein structural changes provides insight into PD pathophysiology and could inform a new concept of structural disease biomarkers. Using limited proteolysis-mass spectrometry (LiP-MS), we identified 76 structurally altered proteins in cerebrospinal fluid (CSF) of individuals with PD relative to healthy donors. These proteins were enriched in processes misregulated in PD, and some proteins also showed structural changes in PD brain samples. CSF protein structural information outperformed abundance information in discriminating between healthy participants and those with PD and improved the discriminatory performance of CSF measures of the hallmark PD protein alpha-synuclein. We also present the first analysis of inter-individual variability of a structural proteome in healthy individuals, identifying biophysical features of variable protein regions. Although independent validation is needed, our data suggest that global analyses of the human structural proteome will guide the development of novel structural biomarkers of disease and enable hypothesis generation about underlying disease processes.ISSN:1545-9993ISSN:1545-998

Global, in situ analysis of the structural proteome in individuals with Parkinson's disease to identify a new class of biomarker

Parkinson’s disease (PD) is a prevalent neurodegenerative disease for which robust biomarkers are needed. Because protein structure reflects function, we tested whether global, in situ analysis of protein structural changes provides insight into PD pathophysiology and could inform a new concept of structural disease biomarkers. Using limited proteolysis–mass spectrometry (LiP–MS), we identified 76 structurally altered proteins in cerebrospinal fluid (CSF) of individuals with PD relative to healthy donors. These proteins were enriched in processes misregulated in PD, and some proteins also showed structural changes in PD brain samples. CSF protein structural information outperformed abundance information in discriminating between healthy participants and those with PD and improved the discriminatory performance of CSF measures of the hallmark PD protein α-synuclein. We also present the first analysis of inter-individual variability of a structural proteome in healthy individuals, identifying biophysical features of variable protein regions. Although independent validation is needed, our data suggest that global analyses of the human structural proteome will guide the development of novel structural biomarkers of disease and enable hypothesis generation about underlying disease processes

p16INK4A-independence of Epstein–Barr virus-induced cell proliferation and virus latency

Epstein-Barr virus (EBV) has the ability to promote cell cycle progression following the initial infection of primary resting B-lymphocytes and to cause cell cycle arrest at the onset of the viral replicative cycle. Various mechanisms have been proposed for the proliferative effects, including the up-regulation of cyclin D2 by the viral EBNA-2 and EBNA-LP proteins, direct binding of EBNA3C to the retinoblastoma protein (pRb), and down-regulation of the p16(INK4A) tumour suppressor by the viral LIMP1 product. To try to gain insight into the relative importance of these mechanisms, the ability of EBV to immortalize lymphocytes from an individual who is genetically deficient for p16(INK4A) was examined. From detailed analyses of the resultant lymphoblastoid cell lines it is concluded that p16(INK4A) status has little bearing on EBV's ability to manipulate the cell cycle machinery and a model to accommodate the previously proposed routes taken by EBV to bypass the restriction point is presented