Real-time monitoring of specific oxygen uptake rates of embryonic stem cells in a microfluidic cell culture device

Oxygen plays a key role in stem cell biology as a signaling molecule and as an indicator of cell energy metabolism. Quantification of cellular oxygen kinetics, i.e. the determination of specific oxygen uptake rates (sOURs), is routinely used to understand metabolic shifts. However current methods to determine sOUR in adherent cell cultures rely on cell sampling, which impacts on cellular phenotype. We present real-time monitoring of cell growth from phase contrast microscopy images, and of respiration using optical sensors for dissolved oxygen. Time-course data for bulk and peri-cellular oxygen concentrations obtained for Chinese hamster ovary (CHO) and mouse embryonic stem cell (mESCs) cultures successfully demonstrated this non-invasive and label-free approach. Additionally, we confirmed non-invasive detection of cellular responses to rapidly changing culture conditions by exposing the cells to mitochondrial inhibiting and uncoupling agents. For the CHO and mESCs, sOUR values between 8 and 60 amol cell(-1) s(-1) , and 5 and 35 amol cell(-1) s(-1) were obtained, respectively. These values compare favorably with literature data. The capability to monitor oxygen tensions, cell growth, and sOUR, of adherent stem cell cultures, non-invasively and in real time, will be of significant benefit for future studies in stem cell biology and stem cell-based therapies

Folklore and traditional ecological knowledge of geckos in Southern Portugal: implications for conservation and science

Traditional Ecological Knowledge (TEK) and folklore are repositories of large amounts of information about the natural world. Ideas, perceptions and empirical data held by human communities regarding local species are important sources which enable new scientific discoveries to be made, as well as offering the potential to solve a number of conservation problems. We documented the gecko-related folklore and TEK of the people of southern Portugal, with the particular aim of understanding the main ideas relating to gecko biology and ecology. Our results suggest that local knowledge of gecko ecology and biology is both accurate and relevant. As a result of information provided by local inhabitants, knowledge of the current geographic distribution of Hemidactylus turcicus was expanded, with its presence reported in nine new locations. It was also discovered that locals still have some misconceptions of geckos as poisonous and carriers of dermatological diseases. The presence of these ideas has led the population to a fear of and aversion to geckos, resulting in direct persecution being one of the major conservation problems facing these animals. It is essential, from both a scientific and conservationist perspective, to understand the knowledge and perceptions that people have towards the animals, since, only then, may hitherto unrecognized pertinent information and conservation problems be detected and resolved

Photoprotection and optimization of sucrose usage contribute to faster recovery of photosynthesis after water deficit at high temperatures in wheat

Plants are increasingly exposed to events of elevated temperature and water deficit, which threaten crop productivity. Understanding the ability to rapidly recover from abiotic stress, restoring carbon assimilation and biomass production, is important to unravel crop climate resilience. This study compared the photosynthetic performance of two Triticum aestivum L. cultivars, Sokoll and Paragon, adapted to the climate of Mexico and UK, respectively, exposed to one week water deficit and high temperatures, in isolation or combination. Measurements included photosynthetic assimilation rate, stomatal conductance, in vitro activities of Rubisco (EC 4.1.1.39) and invertase (INV, EC 3.2.1.26), antioxidant capacity and chlorophyll a fluorescence. In both genotypes, under elevated temperatures and water deficit (WD38℃), the photosynthetic limitations were mainly due to stomatal restrictions and to a decrease in the electron transport rate. Chlorophyll a fluorescence parameters clearly indicate differences between the two genotypes in the photoprotection when subjected to WD38℃ and showed faster recovery of Paragon after stress relief. The activity of the cytosolic invertase (CytINV) under these stress conditions was strongly related to the fast photosynthesis recovery of Paragon. Taken together, the results suggest that optimal sucrose export/utilization and increased photoprotection of the electron transport machinery are important components to limit yield fluctuations due to water shortage and elevated temperatures

Aspectos fitotécnicos e fitoquímicos de acessos de fáfia

The medicinal species fafia (Hebanthe eriantha), is a product of extractivism in the region of the Paraiba Valley, São Paulo state, Brazil, with endangered genetic variability. This study did an agronomical characterization and an analysis of active compounds of five accessions. This research is a partnership of UNESP-Botucatu Medicinal Plants Laboratory, Paraiba Valley Center (APTA) and CPQBA-UNICAMP Agrotechnological Division. A field experiment using completely random blocks with five accessions and seven replications was used. The stem and leaf wet/dry weights, root wet/dry weight, length of the longest stem, foliar area, foam index and pfaffic acid content samples were evaluated. Accession I1800 had root dry weight, stem and leaf dry weight, and foliar area greater than other accessions (176.16 g/plant, 7.301 kg/plant, 155.04 cm² ). Pfaffic acid content was similar to other accessions (0.640-0.366 %mm-1) Among all five accessions, there is a positive correlation among the foliar area and stem and leaf dry weight and between the foliar area and root dry weight.A espécie medicinal fáfia (Hebanthe eriantha), é um recurso extrativista da região do Vale do Paraíba, cuja variabilidade genética encontra-se ameaçada. Neste estudo realizou-se a caracterização agronômica e análise de princípio ativo de cinco acessos. Este estudo é uma parceria entre o Laboratório de Plantas Medicinais da UNESP-Botucatu, o Pólo Vale do Paraíba em Pindamonhangaba da Agência Paulista de Tecnologia dos Agronegócios (APTA) e a Divisão de Agrotecnologia do Centro Pluridisciplinar de Pesquisas Químicas Biológicas e Agrícolas (CPQBA) da UNICAMP. Foi instalado um ensaio experimental a campo em blocos casualizados com cinco acessos e sete repetições. Foram avaliadas a massa fresca e seca da parte aérea, a massa fresca e seca da raiz, o comprimento da maior haste, a área foliar, o índice de espuma e o teor de ácido fáfico das amostras. O acesso I1800 apresentou uma massa seca de raiz, massa seca da parte aérea e área foliar superior aos demais acessos (176,16 g/planta, 7,301 kg/planta, 155,04 cm² ). O teor de ácido fáfico não diferiu entre os acessos (0,640-0,366 %/mm) Existe uma correlação positiva entre área foliar e massa seca da parte aérea e entre área foliar e massa seca de raízes entre os cinco acessos.133138Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Direct metallisation of polyetherimide substrates by activation with different metals

This article reports the performance of different metallic ions and nanoparticles (Ag, Cu, Ni, Pd, Cr, Co, Au and Fe) used as seed layers, formed by chemical or optical reduction, for the electroless Cu plating of metal tracks onto polyetherimide (PEI). Plated Cu performance was tested by adhesion, electrical conductivity, plating rate, XPS, SEM, XRD and EDX analysis. The application of Cu and Ag seeds resulted in high quality electroless Cu deposits presenting strong adhesion properties and high conductivity ((2.0 ± 0.5) × 107 S/m and (3.6 ± 0.2) × 107 S/m, respectively) compared with bulk copper (5.96 × 107 S/m). Performance is attributed to the high surface density and uniformity of seed layers. Of the metals, only Ag ions were photoreduced under the conditions applied and were subsequently used to electroless Cu plate high quality track features of 150 μm width. The application of sulphuric acid pre-treatment to PEI prior to Ag ion exchange, improved the photoinitiated track formation process, as demonstrated by a threefold increase to both photoinduced Ag nanoparticle density on the surface and electroless Cu plating rate, as well as improved electroless Cu adhesion to PEI

A Rapid Photopatterning Method for Selective Plating of 2D and 3D Microcircuitry on Polyetherimide

In this work, a method for the rapid synthesis of metallic microtracks on polyetherimide is presented. The method relies on the photosynthesis of silver nanoparticles on the surface of the polymer substrates from photosensitive silver chloride (AgCl), which is synthesized directly on the polyetherimide surface. The study reveals that the use of AgCl as a photosensitive intermediate accelerates the reactions leading to the formation of silver nanoparticles by up to two orders of magnitude faster than other photodecomposition schemes. The patterning can be conducted under blue light, with notable advantages over UV exposure. Polymers of significant interest to the microelectronics and 3D printing industries can be directly patterned by light using this photography‐inspired technique at throughputs high enough to be commercially advantageous. Light exposures as short as a few seconds are sufficient to allow the direct metallization of the illuminated polyetherimide surface. The results show that the silver required for the seed layer is minimal, and the later copper electroless plating results in the selective growth of conductive tracks for circuitry on the light‐patterned areas, both on flexible films and 3D printed surfaces

A rapid technique for the direct metallization of PDMS substrates for flexible and stretchable electronics applications

Metallization of a polydimethylsiloxane (PDMS)-based substrate is a challenge due to the difficulties in forming crack-free polymer and metal features using standard deposition techniques. Frequently, additional adhesion layers, rigid substrates, multiple processing steps (lift-off and etching) and expensive metal sputtering techniques are required, to achieve such metal patterns. This work presents a novel and rapid technique for the direct metallization of PDMS substrates using photolithography and electroless copper plating. The method has the advantage of not requiring expensive vacuum processing or multiple metallization steps. Electroless copper layer is demonstrated to have a strong adhesion to PDMS substrate with a high conductivity of (3.6 ± 0.7) × 107 S/m, which is close to the bulk copper (5.9 × 107 S/m). The copper-plated PDMS substrate displays mechanical and electrical stability whilst undergoing stretching deformations up to 10% due to applied strain. A functional electronic circuit was fabricated as a demonstration of the mechanical integrity of the copper-plated PDMS after bending

Ontologies relevant to behaviour change interventions: a method for their development [version 2; peer review: 1 not approved]

Background: Behaviour and behaviour change are integral to many aspects of wellbeing and sustainability. However, reporting behaviour change interventions accurately and synthesising evidence about effective interventions is hindered by lacking a shared, scientific terminology to describe intervention characteristics. Ontologies are knowledge structures that provide controlled vocabularies to help unify and connect scientific fields. To date, there is no published guidance on the specific methods required to develop ontologies relevant to behaviour change. We report the creation and refinement of a method for developing ontologies that make up the Behaviour Change Intervention Ontology (BCIO). / Aims: (1) To describe the development method of the BCIO and explain its rationale; (2) To provide guidance on implementing the activities within the development method. / Method and results: The method for developing ontologies relevant to behaviour change interventions was constructed by considering principles of good practice in ontology development and identifying key activities required to follow those principles. The method’s details were refined through application to developing two ontologies. The resulting ontology development method involved: (1) defining the ontology’s scope; (2) identifying key entities; (3) refining the ontology through an iterative process of literature annotation, discussion and revision; (4) expert stakeholder review; (5) testing inter-rater reliability; (6) specifying relationships between entities, and; (7) disseminating and maintaining the ontology. Guidance is provided for conducting relevant activities for each step. / Conclusions: We have developed a detailed method for creating ontologies relevant to behaviour change interventions, together with practical guidance for each step, reflecting principles of good practice in ontology development. The most novel aspects of the method are the use of formal mechanisms for literature annotation and expert stakeholder review to develop and improve the ontology content. We suggest the mnemonic SELAR3, representing the method’s first six steps as Scope, Entities, Literature Annotation, Review, Reliability, Relationships

Representation of behaviour change interventions and their evaluation: Development of the Upper Level of the Behaviour Change Intervention Ontology [version 2; peer review: 1 approved, 1 approved with reservations]

Background: Behaviour change interventions (BCI), their contexts and evaluation methods are heterogeneous, making it difficult to synthesise evidence and make recommendations for real-world policy and practice. Ontologies provide a means for addressing this. They represent knowledge formally as entities and relationships using a common language able to cross disciplinary boundaries and topic domains. This paper reports the development of the upper level of the Behaviour Change Intervention Ontology (BCIO), which provides a systematic way to characterise BCIs, their contexts and their evaluations. Methods: Development took place in four steps. (1) Entities and relationships were identified by behavioural and social science experts, based on their knowledge of evidence and theory, and their practical experience of behaviour change interventions and evaluations. (2) The outputs of the first step were critically examined by a wider group of experts, including the study ontology expert and those experienced in annotating relevant literature using the initial ontology entities. The outputs of the second step were tested by (3) feedback from three external international experts in ontologies and (4) application of the prototype upper-level BCIO to annotating published reports; this informed the final development of the upper-level BCIO. Results: The final upper-level BCIO specifies 42 entities, including the BCI scenario, elaborated across 21 entities and 7 relationship types, and the BCI evaluation study comprising 10 entities and 9 relationship types. BCI scenario entities include the behaviour change intervention (content and delivery), outcome behaviour, mechanism of action, and its context, which includes population and setting. These entities have corresponding entities relating to the planning and reporting of interventions and their evaluations. Conclusions: The upper level of the BCIO provides a comprehensive and systematic framework for representing BCIs, their contexts and their evaluations. Keyword

Development of an Intervention Setting Ontology for behaviour change: Specifying where interventions take place

Background: Contextual factors such as an intervention's setting are key to understanding how interventions to change behaviour have their effects and patterns of generalisation across contexts. The intervention's setting is not consistently reported in published reports of evaluations. Using ontologies to specify and classify intervention setting characteristics enables clear and reproducible reporting, thus aiding replication, implementation and evidence synthesis. This paper reports the development of a Setting Ontology for behaviour change interventions as part of a Behaviour Change Intervention Ontology, currently being developed in the Wellcome Trust funded Human Behaviour-Change Project. Methods: The Intervention Setting Ontology was developed following methods for ontology development used in the Human Behaviour-Change Project: 1) Defining the ontology's scope, 2) Identifying key entities by reviewing existing classification systems (top-down) and 100 published behaviour change intervention reports (bottom-up), 3) Refining the preliminary ontology by literature annotation of 100 reports, 4) Stakeholder reviewing by 23 behavioural science and public health experts to refine the ontology, 5) Assessing inter-rater reliability of using the ontology by two annotators familiar with the ontology and two annotators unfamiliar with it, 6) Specifying ontological relationships between setting entities and 7) Making the Intervention Setting Ontology machine-readable using Web Ontology Language (OWL) and publishing online. Results: The Intervention Setting Ontology consists of 72 entities structured hierarchically with two upper-level classes: Physical setting including Geographic location, Attribute of location (including Area social and economic condition, Population and resource density sub-levels) and Intervention site (including Facility, Transportation and Outdoor environment sub-levels), as well as Social setting. Inter-rater reliability was found to be 0.73 (good) for those familiar with the ontology and 0.61 (acceptable) for those unfamiliar with it. Conclusion: The Intervention Setting Ontology can be used to code information from diverse sources, annotate the setting characteristics of existing intervention evaluation reports and guide future reporting