A transcriptomics approach to study the molecular mechanisms regulating interleukin-10 gene expression in T helper cells

Interleukin-10 (IL-10) is an immunoregulatory cytokine that has a vital role in maintaining a balanced and appropriate immune response. CD4+ T helper (Th) cells are important in regulating an effective immune response, and are a dominant source of IL-10. Despite the different signalling pathways that result in the polarisation of each Th subset and lead to the expression of their hallmark cytokines, IL-10 is expressed by all of the different Th subsets. We show that Th1 cells cultured with IL-12 produce IFNγ and small amounts of IL-10, while Th1 cells cultured with IL-12 and IL-27 produce large amounts of IL-10 and IFNγ. Furthermore, we show that Th17 cells can produce IL-10, or not, depending on the presence of IL-2. However, these Th cell populations are phenotypically heterogeneous, particularly with respect production of IL-10 protein. Less than half of each of these in vitro cultured Th cell populations expresses IL-10 or the hallmark cytokine, and co-expression of IL-10 and the hallmark cytokine is also heterogeneous. Therefore we devised and implemented an innovative new technique that enables RNA-Seq analysis of different intracellular cytokine producing cell subpopulations from within Th subsets. We find that it is possible to extract high quality mRNA from Th samples, even though they have been fixed and stained for intracellular cytokines, and that the data from these samples is replicable. Using this technique we have identified potential molecules and pathways by which (I) IL-27 drives IFNγ and IL-10 production by Th1 cells, and (II) IL-2 drives IL-10 production by Th17 cells. Furthermore, by separating different intracellular cytokine producing subpopulations within different Th1 and Th17 cell subsets we have found that cytokine producing and non-cytokine producing subpopulations within heterogeneous Th subsets have significantly different transcriptional profiles and that some pathways and molecules are enriched in IL-10 producing cells

An Origin-of-Life Reactor to Simulate Alkaline Hydrothermal Vents

Chemiosmotic coupling is universal: practically all cells harness electrochemical proton gradients across membranes to drive ATP synthesis, powering biochemistry. Autotrophic cells, including phototrophs and chemolithotrophs, also use proton gradients to power carbon fixation directly. The universality of chemiosmotic coupling suggests that it arose very early in evolution, but its origins are obscure. Alkaline hydrothermal systems sustain natural proton gradients across the thin inorganic barriers of interconnected micropores within deep-sea vents. In Hadean oceans, these inorganic barriers should have contained catalytic Fe(Ni)S minerals similar in structure to cofactors in modern metabolic enzymes, suggesting a possible abiotic origin of chemiosmotic coupling. The continuous supply of H2 and CO2 from vent fluids and early oceans, respectively, offers further parallels with the biochemistry of ancient autotrophic cells, notably the acetyl CoA pathway in archaea and bacteria. However, the precise mechanisms by which natural proton gradients, H2, CO2 and metal sulphides could have driven organic synthesis are uncertain, and theoretical ideas lack empirical support. We have built a simple electrochemical reactor to simulate conditions in alkaline hydrothermal vents, allowing investigation of the possibility that abiotic vent chemistry could prefigure the origins of biochemistry. We discuss the construction and testing of the reactor, describing the precipitation of thin-walled, inorganic structures containing nickel-doped mackinawite, a catalytic Fe(Ni)S mineral, under prebiotic ocean conditions. These simulated vent structures appear to generate low yields of simple organics. Synthetic microporous matrices can concentrate organics by thermophoresis over several orders of magnitude under continuous open-flow vent conditions

Uncovering human needs through visual research methods: Two commercial case studies

This paper presents two respective case studies which demonstrate how visual research methods can elicit a deep understanding of the needs of potential end users and drive product and service development at a strategic business level.The engagement of users in the development of products, services and systems has been explored by a number of design disciplines in the last few decades including but not limited to product design, human-computer interaction, systems design and service design. Each has recognised the importance of understanding the humans who will potentially be using their design outcome. Notable research methods include ethnographic inspired research, in-context and lab-based observations, interviews and the user trial of prototypes. However, these approaches also have their critics and limitations ranging from the need for incremental adjustment rather than radical design, being time-consuming and costly processes, and the large volume of ‘messy data’ being collected contributing to the complexities of ‘wicked problems’.In response to some of these limitations, a number of research methods have emerged which are more arts-based in nature i.e. the act of creating allows the researcher to extract ‘deeper’ human needs (tacit and latent needs) in a drastically shorter timescale. To fully utilise such approaches it is essential that a study be designed which amalgamates diverse research methods. The two case studies presented in this paper employ a variety of traditional and generative design research methods in live commercial projects. The specific project outcomes are retained under Intellectual Property and, as such, this paper critically focuses on the value of the process and methods utilised, their relationship to the wider concept of arts-based research, and discusses issues related to their application in commercial work

Language matters. Addressing the use of language in the care of people with diabetes: position statement of the English Advisory Group

The language used by healthcare professionals can have a profound impact on how people living with diabetes, and those who care for them, experience their condition and feel about living with it day-to-day. At its best, good use of language, both verbal and written, can lower anxiety, build confidence, educate and help to improve self-care. Conversely, poor communication can be stigmatizing, hurtful and undermining of self-care and can have a detrimental effect on clinical outcomes. The language used in the care of those with diabetes has the power to reinforce negative stereotypes, but it also has the power to promote positive ones. The use of language is controversial and has many perspectives. The development of this position statement aimed to take account of these as well as the current evidence base. A working group, representing people with diabetes and key organizations with an interest in the care of people with diabetes, was established to review the use of language. The work of this group has culminated in this position statement for England. It follows the contribution of Australia and the USA to this important international debate. The group has set out practical examples of language that will encourage positive interactions with those living with diabetes and subsequently promote positive outcomes. These examples are based on a review of the evidence and are supported by a simple set of principles

Near-Infrared Laser Adjuvant for Influenza Vaccine

Safe and effective immunologic adjuvants are often essential for vaccines. However, the choice of adjuvant for licensed vaccines is limited, especially for those that are administered intradermally. We show that non-tissue damaging, near-infrared (NIR) laser light given in short exposures to small areas of skin, without the use of additional chemical or biological agents, significantly increases immune responses to intradermal influenza vaccination without augmenting IgE. The NIR laser-adjuvanted vaccine confers increased protection in a murine influenza lethal challenge model as compared to unadjuvanted vaccine. We show that NIR laser treatment induces the expression of specific chemokines in the skin resulting in recruitment and activation of dendritic cells and is safe to use in both mice and humans. The NIR laser adjuvant technology provides a novel, safe, low-cost, simple-to-use, potentially broadly applicable and clinically feasible approach to enhancing vaccine efficacy as an alternative to chemical and biological adjuvants

Eastern philosophies of education : Buddhist, Hindu, Daoist, and Confucian readings of Plato’s cave

This chapter provides readers with an understanding of some basic principles of selected Eastern traditions and their relation to philosophy of education. The attempt to characterize such diverse traditions and understandings of education raises numerous hermeneutical issues which can only be addressed through a pedagogical reduction as a vehicle for understanding. In this case, we have employed Plato’s cave allegory as that methodological and pedagogical vehicle. We explore aspects of the ontology, epistemology, and ethics of Buddhist, Hindu (focused on classical yoga), Daoist, and Confucian traditions, interpreting elements from Plato’s allegory in order to throw light onto the educational ideas and implications of those Eastern traditions

The Rachel Carson Letters and the Making of Silent Spring

Environment, conservation, green, and kindred movements look back to Rachel Carson’s 1962 book Silent Spring as a milestone. The impact of the book, including on government, industry, and civil society, was immediate and substantial, and has been extensively described; however, the provenance of the book has been less thoroughly examined. Using Carson’s personal correspondence, this paper reveals that the primary source for Carson’s book was the extensive evidence and contacts compiled by two biodynamic farmers, Marjorie Spock and Mary T. Richards, of Long Island, New York. Their evidence was compiled for a suite of legal actions (1957-1960) against the U.S. Government and that contested the aerial spraying of dichlorodiphenyltrichloroethane (DDT). During Rudolf Steiner’s lifetime, Spock and Richards both studied at Steiner’s Goetheanum, the headquarters of Anthroposophy, located in Dornach, Switzerland. Spock and Richards were prominent U.S. anthroposophists, and established a biodynamic farm under the tutelage of the leading biodynamics exponent of the time, Dr. Ehrenfried Pfeiffer. When their property was under threat from a government program of DDT spraying, they brought their case, eventually lost it, in the process spent US$100,000, and compiled the evidence that they then shared with Carson, who used it, and their extensive contacts and the trial transcripts, as the primary input for Silent Spring. Carson attributed to Spock, Richards, and Pfeiffer, no credit whatsoever in her book. As a consequence, the organics movement has not received the recognition, that is its due, as the primary impulse for Silent Spring, and it is, itself, unaware of this provenance

Co-design, evaluation and the Northern Ireland Innovation Lab

Around the world there are more than 100 policy labs—multi-disciplinary government teams developing public services and policies using innovation methods to engage citizens and stakeholders. These policy labs use a range of innovation methods and approaches, including co-production, co-creation, co-design, behavioural insights, systems thinking, ethnography, data science, nudge theory and lean processes. Although the methods may vary, one element is consistent: policy labs actively, creatively and collaboratively engage the public and a wide range of stakeholders in jointly developing solutions. The Northern Ireland Public Sector Innovation Lab (iLab) is part of a growing UK and international community of policy labs using co-design to engage with users for value co-creation, aiming to improve public governance by creating a safe space to generate ideas, test prototypes and refine concepts with beneficiaries. Drawing on iLab’s experience, this paper explores three questions: What are the main determinants of effective co-design? What are the unintended consequences of co-design? And what lessons can be learned from iLab and shared with other policy labs

Evaluation of antiarthritic activity of Strychnos potatorum Linn seeds in Freund's adjuvant induced arthritic rat model

<p>Abstract</p> <p>Background</p> <p><it>Strychnos potatorum </it>Linn (Loganiaceae) is a moderate sized tree found in southern and central parts of India, Sri Lanka and Burma. In traditional system of medicine, <it>Strychnos potatorum </it>Linn seeds were used for various ailments including inflammation, diabetes etc. To investigate the folkloric use of the seeds the present study was carried out on Freund's adjuvant induced arthritic rats.</p> <p>Methods</p> <p>The present study states the effect of the aqueous extract (SPE) and the whole seed powder (SPP) of <it>Strychnos potatorum </it>Linn seeds on the Freund's complete adjuvant (FCA) induced arthritic rat paw edema, body weight changes and alterations in haematological and biochemical parameters in both developing and developed phases of arthritis. Histopathology of proximal interphalangeal joints and radiology of hind legs were studied.</p> <p>Results</p> <p>In FCA induced arthritic rats, there was significant increase in rat paw volume and decrease in body weight increment, whereas SPP and SPE treated groups, showed significant reduction in paw volume and normal gain in body weight. The altered haematological parameters (Hb, RBC, WBC and ESR) and biochemical parameters (blood urea, serum creatinine, total proteins and acute phase proteins) in the arthritic rats were significantly brought back to near normal by the SPP and SPE treatment at the dose of 200 mg/kg/p.o in both developing and developed phases of arthritis. Further the histopathological and radiological studies revealed the antiarthritic activity of SPP and SPE by indicating fewer abnormalities in these groups when compared to the arthritic control group.</p> <p>Conclusion</p> <p>In conclusion, both SPP and SPE at the specified dose level of 200 mg/kg, p.o. showed reduction in rat paw edema volume and it could significantly normalize the haematological and biochemical abnormalities in adjuvant induced arthritic rats in both developing and developed phases of FCA induced arthritis. Further the histopathological and radiological studies confirmed the antiarthritic activity of SPP and SPE.</p

c-Maf controls immune responses by regulating disease-specific gene networks and repressing IL-2 in CD4+ T cells

The transcription factor c-Maf induces the anti-inflammatory cytokine IL-10 in CD4+ T cells in vitro. However, the global effects of c-Maf on diverse immune responses in vivo are unknown. Here we found that c-Maf regulated IL-10 production in CD4+ T cells in disease models involving the TH1 subset of helper T cells (malaria), TH2 cells (allergy) and TH17 cells (autoimmunity) in vivo. Although mice with c-Maf deficiency targeted to T cells showed greater pathology in TH1 and TH2 responses, TH17 cell–mediated pathology was reduced in this context, with an accompanying decrease in TH17 cells and increase in Foxp3+ regulatory T cells. Bivariate genomic footprinting elucidated the c-Maf transcription-factor network, including enhanced activity of NFAT; this led to the identification and validation of c-Maf as a negative regulator of IL-2. The decreased expression of the gene encoding the transcription factor RORγt (Rorc) that resulted from c-Maf deficiency was dependent on IL-2, which explained the in vivo observations. Thus, c-Maf is a positive and negative regulator of the expression of cytokine-encoding genes, with context-specific effects that allow each immune response to occur in a controlled yet effective manner