Barium alginate capsules for 3D immobilisation of living cells: morphology, membrane properties and permeability

Encapsulation in a barium alginate membrane is a promising strategy to obtain a three dimensional culture of living cells: membrane properties are crucial for a realistic clinical application. A one-step encapsulation technique, recently developed for controlled release of boar semen, was employed to prepare barium alginate and protamine-alginate membranes: permeability to two model molecules (haemoglobin and glucose) was evaluated. Capsules were evaluated for technological properties and scanning electron microscopy was used to examine the external morphology of the capsules and the 3D distribution of the cells within the core. The results indicate that 3D arrangement and cell shape are maintained, capsule dimensions and mechanical properties can be modulated, as well as their permeability to model molecules such as haemoglobin and glucose

Transient transfection of porcine granulosa cells after 3D culture in barium alginate capsules

Three-dimensional culture systems in barium alginate capsules can be employed to maintain primary granulosa cells in an undifferentiated state for almost 6 days. This is due to a self-organization of cells in a pseudofollicular structure. The transfection of primary granulosa cells is a necessary condition when employing these culture systems for several purposes, for example as an in vitro toxicity test or the development of oocytes or zygotes. In this work, the feasibility of two transient transfection techniques (liposome-mediated and electroporation) was assessed in primary porcine granulosa cells after a 6-day culture in an artificial extracellular matrix (barium alginate membrane). Human recombinant green fluorescent protein was chosen as a molecular readout, and protein expression was assessed after 48 hours from transfection. Liposome-mediated transfection gave low transfection levels, with increasing yields from 2 to 12 microgDNA/ml of medium; the maximum percentage (85.7%) was reached at 12 microgDNA/ml of medium. Electroporation-mediated transfection yields were higher: the best results (81.7% of transfected cells) were achieved with two 50V pulses and 12 microg/ml DNA. The application of a single or double pulse (50V) at 4 mgDNA/ml gave negligible results. These results indicate that primary granulosa cell cultured in barium alginate capsules can be transfected by electroporation with high transfection yields

VRP-REP: a vehicle routing community repository

International audienc

Immunocytochemical Characterization of Alzheimer’s Disease Hallmarks in APP/PS1 Transgenic Mice Treated with a New Anti-Amyloid-β Vaccine

Introduction: APP/PS1 double-transgenic mouse models of Alzheimer’s disease (AD), which overexpress mutated forms of the gene for the human amyloid precursor protein (APP) and presenilin 1 (PS1), have provided robust neuropathological hallmarks of an AD-like pattern at early ages. This study aimed to characterize immunocytochemical patterns of the AD mouse brain, which is treated with the EB101 vaccine, as a model for human AD.Material and methods: In this novel vaccine, a new approach has been taken to circumvent past failures with A? vaccines by judiciously selecting an adjuvant consisting of a physiological matrix embedded in liposomes, composed of naturally occurring phospholipids (phosphatidylcholine, phosphatidylglycerol, and cholesterol).Results: Our findings showed that the administration of amyloid-?1?42 (A?) and sphingosine-1-phosphate emulsified in liposome complex (EB101) to APP/PS1 mice before the onset of A? brain deposition (at 7 weeks of age) and/or at an older age (35 weeks of age) can be effective in both halting the progression and clearing the AD-like neuropathological hallmarks. In addition, passive immunization with EB101 did not activate inflammatory responses from the immune system and astrocytes. Consistent with a decreased inflammatory background, the basal immunological interaction between the T cells and the affected areas (hippocampus) in the brain of treated mice was notably reduced.Conclusion: These results provide strong evidence that immunization with the EB101 vaccine prevents and attenuates AD neuropathology in this type of double-transgenic mice

Determination of fatty acids profile in original brown cows dairy products and relationship with alpine pasture farming system

This study aimed to evaluate the relationships between fatty acids and the pattern that most contributes to discriminate between two farming systems, in which the main difference was the practice, or not, of alpine summer-grazing. Milk and cheese were sampled every month in two farms of Original Brown cows identical under geographical location and management during no grazing season point of view in the 2018 season. Fatty acids concentrations were determined by gas chromatography. The principal component analysis extracted three components (PCs). Mammary gland de novo synthetized fatty acids (C14:0, C14:1 n9, and C16:0) and saturated and monosaturated C18 fatty acids (C18:0, C18:1 n9c) were inversely associated in the PC1; PC2 included polyunsaturated C18 fatty acids (C18:2 n6c, C18:3 n3) and C15:0 while conjugated linoleic acid (CLA n9c, n11t) and fatty acids containing 20 or more carbon atoms (C21:0, C20:5 n3) were associated in the PC3. The processes of rumen fermentation and de novo synthesis in mammary gland that are, in turn, influenced by diet, could explain the relationships between fatty acids within each PC. The discriminant analyses showed that the PC2 included the fatty acids profile that best discriminated between the two farming systems, followed by PC3 and, lastly, PC1. This model, if validated, could be an important tool to the dairy industry

Disease Burden and Government Spending On Mental, Neurological, And Substance Use Disorders, and Self-Harm: Cross-Sectional, Ecological Study of Health System Response in the Americas

Background Disorders affecting mental health are highly prevalent, can be disabling, and are associated with substantial premature mortality. Yet national health system responses are frequently under-resourced, inefficient, and ineffective, leading to an imbalance between disease burden and health expenditures. We estimated the disease burden in the Americas caused by disorders affecting mental health. This measure was adjusted to include mental, neurological, and behavioural disorders that are frequently not included in estimates of mental health burden. We propose a framework for assessing the imbalance between disease burden and health expenditures. Methods In this cross-sectional, ecological study, we extracted disaggregated disease burden data from the Global Health Data Exchange to produce country-level estimates for the proportion of total disease burden attributable to mental disorders, neurological disorders, substance use disorders, and self-harm (MNSS) in the Americas. We collated data from the WHO Assessment Instrument for Mental Health Systems and the WHO Mental Health Atlas on country-level mental health spending as a proportion of total government health expenditures, and of psychiatric hospital spending as a proportion of mental health expenditures. We used a metric capturing the imbalance between disease burden and mental health expenditures, and modelled the association between this imbalance and real (ie, adjusted for purchasing power parity) gross domestic product (GDP). Findings Data were collected from July 1, 2016, to March 1, 2017. MNSS comprised 19% of total disability-adjusted lifeyears in the Americas in 2015. Median spending on mental health was 2·4% (IQR 1·3–4·1) of government health spending, and median allocation to psychiatric hospitals was 80% (52–92). This spending represented an imbalance in the ratio between disease burden and efficiently allocated spending, ranging from 3:1 in Canada and the USA to 435:1 in Haiti, with a median of 32:1 (12–170). Mental health expenditure as a proportion of government health spending was positively associated with real GDP (β=0·68 [95% CI 0·24–1·13], p=0·0036), while the proportion allocated to psychiatric hospitals (β=–0·5 [–0·79 to –0·22], p=0·0012) and the imbalance in efficiently allocated spending (β=–1·38 [–1·97 to –0·78], p=0·0001) were both inversely associated with real GDP. All estimated coefficients were significantly different from zero at the 0·005 level. Interpretation A striking imbalance exists between government spending on mental health and the related disease burden in the Americas, which disproportionately affects low-income countries and is likely to result in undertreatment, increased avoidable disability and mortality, decreased national economic output, and increased household-level health spending

Pentraxin 3 is up-regulated in epithelial mammary cells during Staphylococcus aureus intra-mammary infection in goat

Pentraxin 3 is the prototypic long pentraxin and is produced by different cell populations (dendritic cells, monocytes/macrophages, endothelial cells, and fibroblasts) after pro-inflammatory stimulation. Different studies demonstrated the up-regulation of PTX3 during mastitis in ruminants, but its role is still unknown. We first investigated the conservation of PTX3 sequence among different species and its pattern of expression in a wide panel of organs from healthy goats. We studied the role modulation of PTX3 during natural and experimental mammary infection, comparing its expression in blood, milk and mammary tissues from healthy and Staphylococcus aureus infected animals. We confirmed the high conservation of the molecule among the different species. Goat PTX3 was expressed at high levels in bone marrow, mammary gland, aorta, rectum, pancreas, skin and lungs. PTX3 was up-regulated in epithelial mammary cells and in milk cells after S. aureus infection, suggesting that it represents a first line of defense in goat udder

Advances in the application and utility of subseasonal-to-seasonal predictions

The joint WWRP–WCRP Subseasonal to Seasonal Prediction Project (e.g., Robertson et al. 2014) created a global repository of experimental or operational near-real-time S2S forecasts and reforecasts (hindcasts) from 11 international meteorological institutions, cohosted by ECMWF and CMA (Vitart et al. 2017). These data are publicly accessible by researchers and users (https://apps.ecmwf.int/datasets/data/s2s and http://s2s.cma.cn/index). With the exception of the fourth case study, which uses GloSea5 forecasts (MacLachlan et al. 2015), all case studies use selected S2S forecasts and reforecasts that are available from this repository, providing a consistent basis for S2S forecast skill assessment and evaluation of their utility.The subseasonal-to-seasonal (S2S) predictive time scale, encompassing lead times ranging from 2 weeks to a season, is at the frontier of forecasting science. Forecasts on this time scale provide opportunities for enhanced application-focused capabilities to complement existing weather and climate services and products. There is, however, a “knowledge–value” gap, where a lack of evidence and awareness of the potential socioeconomic benefits of S2S forecasts limits their wider uptake. To address this gap, here we present the first global community effort at summarizing relevant applications of S2S forecasts to guide further decision-making and support the continued development of S2S forecasts and related services. Focusing on 12 sectoral case studies spanning public health, agriculture, water resource management, renewable energy and utilities, and emergency management and response, we draw on recent advancements to explore their application and utility. These case studies mark a significant step forward in moving from potential to actual S2S forecasting applications. We show that by placing user needs at the forefront of S2S forecast development—demonstrating both skill and utility across sectors—this dialogue can be used to help promote and accelerate the awareness, value, and cogeneration of S2S forecasts. We also highlight that while S2S forecasts are increasingly gaining interest among users, incorporating probabilistic S2S forecasts into existing decision-making operations is not trivial. Nevertheless, S2S forecasting represents a significant opportunity to generate useful, usable, and actionable forecast applications for and with users that will increasingly unlock the potential of this forecasting time scale.DD gratefully acknowledges support from the Swiss National Science Foundation through project PP00P2_170523. For case study 1, ACP and WTKH were funded by the U.K. Climate Resilience Programme, supported by the UKRI Strategic Priorities Fund. RWL was funded by NERC Grant NE/P00678/1 and by the BER DOE Office of Science Federal Award DE-SC0020324. TS was funded by NERC Independent Research Fellowship (NE/P018637/1). CMG and DB were funded by the Helmholtz Young Investigator Group “SPREADOUT” Grant VH-NG-1243. Case study 2 was supported by the U.K. Global Challenges Research Fund NE/P021077/1 (GCRF African SWIFT) and the Tertiary Education Trust Fund (TETFUND) of Nigeria TETFund/DR&D/CE/NRF/STI/73/VOL.1. EO thanks Adrian Tomkins of ICTP, Italy, for his contribution. Case study 3 was undertaken as part of the Columbia World Project, ACToday, Columbia University (https://iri.columbia.edu/actoday/). Case study 4 was supported by the ForPAc (Towards Forecast-based Preparedness Action) project within the NERC/FCDO SHEAR Programme NE/P000428/1, NE/P000673/1, and NE/P000568/1. Case study 5 was undertaken as part of the International Research Applications Project, funded by the U.S. National Oceanic and Atmospheric Administration. EO thanks IRAP project colleagues at The University of Arizona, Indian Meteorological Department, Regional Integrated Multi-Hazard Early Warning System for Africa and Asia, and two of Bihar’s State Agricultural Universities for their contributions. For case study 6, CASC thanks Conselho Nacional de Desenvolvimento Científico e Tecnológico Process 305206/2019-2 and Fundação de Amparo à Pesquisa do Estado de São Paulo Process 2015/50687-8 (CLIMAX Project) for their support. For case study 7, DW’s contributions were carried out under contract with the National Aeronautics and Space Administration. Case study 8 was funded by the EU Horizon 2020 Research and Innovation Programme Grant 7767874 (S2S4E). We also acknowledge the Subseasonal-to-Seasonal Project’s Real-Time Pilot Initiative for providing access to real-time forecasts. For case study 9, TIC-LCPE Hydro-04 was funded by the University of Strathclyde’s Low Carbon Power and Energy program. JB was supported by EPSRC Innovation Fellowship EP/R023484/1. We thank Andrew Low and Richard Hearnden from SSE Renewables for their input. Case study 10 was supported by the Earth Systems and Climate Change Hub under the Australian Government’s National Environmental Science Program, and the Decadal Climate Forecasting Project (CSIRO). Case study 11 was funded by the Technologies for Sustainable Built Environments Centre, Reading University, in conjunction with the EPSRC Grant EP/G037787/1 and BT PLC. Case study 12 was funded through the framework service contract for operating the EFAS Computational Center Contract 198702 and the Copernicus Fire Danger Computations Contract 389730 295 in support of the Copernicus Emergency Management Service and Early Warning Systems between the Joint Research Centre and ECMWF.Peer Reviewed"Article signat per 60 autors/es: Christopher J. White, Daniela I. V. Domeisen, Nachiketa Acharya, Elijah A. Adefisan, Michael L. Anderson, Stella Aura, Ahmed A. Balogun, Douglas Bertram, Sonia Bluhm, David J. Brayshaw, Jethro Browell, Dominik Büeler, Andrew Charlton-Perez, Xandre Chourio, Isadora Christel, Caio A. S. Coelho, Michael J. DeFlorio, Luca Delle Monache, Francesca Di Giuseppe, Ana María García-Solórzano, Peter B. Gibson, Lisa Goddard, Carmen González Romero, Richard J. Graham, Robert M. Graham, Christian M. Grams, Alan Halford, W. T. Katty Huang, Kjeld Jensen, Mary Kilavi, Kamoru A. Lawal, Robert W. Lee, David MacLeod, Andrea Manrique-Suñén, Eduardo S. P. R. Martins, Carolyn J. Maxwell, William J. Merryfield, Ángel G. Muñoz, Eniola Olaniyan, George Otieno, John A. Oyedepo, Lluís Palma, Ilias G. Pechlivanidis, Diego Pons, F. Martin Ralph, Dirceu S. Reis Jr., Tomas A. Remenyi, James S. Risbey, Donald J. C. Robertson, Andrew W. Robertson, Stefan Smith, Albert Soret, Ting Sun, Martin C. Todd, Carly R. Tozer, Francisco C. Vasconcelos Jr., Ilaria Vigo, Duane E. Waliser, Fredrik Wetterhall, and Robert G. Wilson"Postprint (author's final draft

Listen to Nice

In describing Humphrey Jennings’ wartime documentary propaganda film, 'Listen to Britain' (1942), a film with an overtly poetic sensibility and dominantly musical soundtrack, John Corner asserts that ‘through listening to Britain, we are enabled to properly look at it'. This idea of sound leading our attention to the images has underpinned much of the collaborative work between composer and sound designer, Geoffrey Cox, and documentary filmmaker, Keith Marley. It is in this context that the article will analyse an extract of A Film About Nice (Marley and Cox 2010), a contemporary re-imagining of Jean Vigo’s silent documentary, 'A propos de Nice' (1930). Reference will be made throughout to the historical context, and the filmic and theoretical influences that have informed the way music and creative sound design have been used to place emphasis on hearing a place, as much as seeing it