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Supplementary Figure 1 from Echinomycin, a Small-Molecule Inhibitor of Hypoxia-Inducible Factor-1 DNA-Binding Activity

The identification of small molecules that inhibit the sequence-specific binding of transcription factors to DNA is an attractive approach for regulation of gene expression. Hypoxia-inducible factor-1 (HIF-1) is a transcription factor that controls genes involved in glycolysis, angiogenesis, migration, and invasion, all of which are important for tumor progression and metastasis. To identify inhibitors of HIF-1 DNA-binding activity, we expressed truncated HIF-1α and HIF-1β proteins containing the basic-helix-loop-helix and PAS domains. Expressed recombinant HIF-1α and HIF-1β proteins induced a specific DNA-binding activity to a double-stranded oligonucleotide containing a canonical hypoxia-responsive element (HRE). One hundred twenty-eight compounds previously identified in a HIF-1–targeted cell-based high-throughput screen of the National Cancer Institute 140,000 small-molecule library were tested in a 96-well plate ELISA for inhibition of HIF-1 DNA-binding activity. One of the most potent compounds identified, echinomycin (NSC-13502), a small-molecule known to bind DNA in a sequence-specific fashion, was further investigated. Electrophoretic mobility shift assay experiments showed that NSC-13502 inhibited binding of HIF-1α and HIF-1β proteins to a HRE sequence but not binding of the corresponding proteins to activator protein-1 (AP-1) or nuclear factor-κB (NF-κB) consensus sequences. Interestingly, chromatin immunoprecipitation experiments showed that NSC-13502 specifically inhibited binding of HIF-1 to the HRE sequence contained in the vascular endothelial growth factor (VEGF) promoter but not binding of AP-1 or NF-κB to promoter regions of corresponding target genes. Accordingly, NSC-13502 inhibited hypoxic induction of luciferase in U251-HRE cells and VEGF mRNA expression in U251 cells. Our results indicate that it is possible to identify small molecules that inhibit HIF-1 DNA binding to endogenous promoters.

Supplementary Figure 2 from Echinomycin, a Small-Molecule Inhibitor of Hypoxia-Inducible Factor-1 DNA-Binding Activity

In recent decades, out-migration has become a key livelihood strategy for many rural households in the middle hills region of Nepal. In this region, rural communities are key actors in the management of local resources such as community forests. Analysis of the link between community forests and out-migration is largely missing in the literature, even though the demographic changes associated with out-migration affect forest resource use and management. This article discusses how rural communities and traditional landscapes in the middle hills are changing following out-migration and how this process has changed the management by rural communities of local forests (including community forests). Overall, the research found a reduced dependency by households on forest products (e.g. firewood, fodder and timber) sourced from community forests. Also, the reduced demand is being supplied increasingly from trees/forests grown on private farmland, including natural regenerated forests and trees planted on abandoned farmland. The declining need for forest products and the lack of an economic incentive for active forest management coupled with a decreasing sense of community has reduced the interest in community forests, leading to less intensive and infrequent forest management. Given the prevalence of out-migration and the changing socio-economic context in the middle hills, it appears time to reconsider the management of community forests beyond a narrow range of uses to enable greater commercialization and encourage ecosystem services to be harnessed so community forests better align with contemporary rural livelihoods and landscapes.

We offer a unified conceptual, behavioral, and econometric framework for optimal fundraising that deals with both synergies and discrepancies between approaches from Economics, Marketing, Psychology, and Sociology. The purpose is to offer a framework that can bridge differences and open a dialogue between disciplines in order to facilitate optimal fundraising design. The literature is extensive, and our purpose is to offer a brief background and perspective on each of the approaches, provide an integrated framework leading to new insights, and discuss areas of future research.

Should advertisers change their message (what is said), just as they do the execution (how it is said) to reflect changing consumer preferences? This paper is the first to quantify how these ad components and their interplay affect brand sales. The authors define the concepts of Market Consistency and changes in ad executions and show how they interact with each other and with the brand’s age in their sales outcome. The empirical analysis confirms the hypotheses in the U.S. minivan market. As a brand matures, executional variations become increasingly beneficial, but changing advertising messaging to remain consistent with customer preferences becomes less effective. For older brands with little executional variation, changing the ad message even reduces sales.The authors thus uncover important boundary conditions for the opposing theories that brands should ‘stick with their message’ versus ‘change with the times’ and advice how to manage advertising as the brand matures.

The conventional approach to urban planning in Thailand has been a centralised process that relies mainly on decisions by urban planners, based on national regulations and policies. This approach has been ineffective in Wat Ket, a small and unique area in Chiang Mai City in northern Thailand. A key problem is that some parts of Wat Ket were designated as a commercial zone, without the involvement and agreement of local residents, leading to land use conflicts and efforts by local people to challenge the zoning. Collaborative urban planning has been advocated and applied in a number of countries over at least the last two decades, but rarely, if at all, in ‘developing’ countries. The main purpose of this paper is to discuss efforts, by local stakeholders, to apply collaborative urban planning as an alternative approach to urban planning in Wat Ket. In support of the collaborative urban planning experiment, land use analysis applying aerial photography and Geographical Information Systems (GIS) was used as a method for quantifying land use changes. Findings from this study are that: (1) the experiment in collaborative urban planning led to some revisions of the land use zoning and the final draft of a specific plan in two designated areas; (2) the potential for collaborative urban planning in strengthening the participation of multiple stakeholders and contributing to knowledge in urban planning was demonstrated; (3) the willingness of the planning agencies to adopt the approach requires time and resources for any change in urban land use planning policy. It is suggested that further documented experiments in collaborative land use planning will be needed to encourage policy change.

Environmental and development discourses in China can be categorised into three narrative motifs framing human–nature relationships: peasant, indigenous, and community. Indigenous and community narratives have been widely adopted by environmental NGOs (eNGOs) in China in promoting community-based natural resource management projects, but there has been very limited critical research on such phenomena. Analysis of socio-economic change in two ethnic minority communities in Yunnan shows that neither narrative theme is fully internalised by the relevant communities. Instead narratives may be strategically modified or even rejected by local communities. This is due to different agendas being held by local communities and eNGOs, and two factors pertinent to rural China: the incompatibility of concepts of ‘community’ in Chinese and international contexts results in confusion, and a lack of recent territorial and cultural claims by rural communities since the collectivist era makes it difficult to construct the identity of a community. It remains challenging for eNGOs in China to advocate either community or indigenous narratives in contexts of rapid socio-economic change.

Background In 1980, smallpox disease was eradicated from nature and Variola virus, the etiological agent of smallpox, was confined to two laboratories, one located in Russia (Moscow) later moved to VECTOR (Novosibirsk, Siberia) and one in the United States (CDC Atlanta). Vaccinations among the general public ceased shortly after the successful eradication campaign, resulting in an increasingly immunologically susceptible population. Because of the possibility of intentional reintroduction of Variola virus and the emergence of other pathogenic poxviruses, there is a great need for the development of medical countermeasures to treat poxvirus disease. It is highly likely that the U.S. FDA “animal rule” will be necessary for regulatory approval of these interventions. Therefore, relevant animal models and the associated supporting assays will require development to stand up to regulatory scrutiny. Methods An optimized real time PCR assay for the detection of orthopoxviruses has been developed by researchers at the United States Army Research Institute of Infectious Diseases (USAMRIID). To support animal studies that will be used to support approval of medical countermeasures by the U.S. FDA, the assay was designed to quantitate poxvirus genomic DNA in a nonhuman primate (cynomolgus macaque) blood matrix as a measurement of viremia. This manuscript describes the validation of the process, including DNA extraction from whole blood anticoagulated with EDTA, for obtaining and quantitating monkeypox genomes by evaluating precision, accuracy, the standard curve, specificity, robustness and stability of the assay and/or components of the assay. Results The assay had a lower limit of quantitation of 50 genome copies/5 uL sample, upper limit of quantitation of 5 × 107 GC/5uL sample and a limit of detection of 2.5 genome copies /5uL sample. The assay was specific for orthopoxvirus. Matrix effects were detected and suggest the presence of PCR inhibitor(s) that was co-extracted with the target DNA. Conclusions The assay has been validated for the purpose of quantitating monkeypox viral load in blood from cynomolgus macaques. This assay has and will continue to support submissions to the FDA for approval of antiviral therapeutics for smallpox. Electronic supplementary material The online version of this article (10.1186/s12985-017-0880-8) contains supplementary material, which is available to authorized users.

Continuous manufacturing (CM) is an emerging technology in the pharmaceutical manufacturing sector, and the understanding of the impact on product quality is currently evolving. As the final purification and isolation step, crystallization has a significant impact on the final physicochemical properties of drug substance and is considered a critical process step in achieving the continuous manufacturing of drug substances. Although many publications previously focused on various innovative techniques to continuously make crystals with desired properties, engineering difficulties such as system design, automation, and integration with process analytical technology (PAT) tools have not been thoroughly discussed. Here, we focus on how to develop a continuous crystallization system, from the perspective of process engineering, and the related risk considerations on product quality. Specifically, we designed and built an automated two-stage mixed suspension mixed product removal (MSMPR) crystallization platfor...

For continuous manufacturing of pharmaceuticals, understanding the dynamics of how a material flows through the process is critical with respect to the development of a control strategy for product quality assurance. Such understanding of the process dynamics can be obtained by characterization of the residence time distribution (RTD). The RTD for a process is not fixed and can vary due to changes in operating conditions or physiochemical properties of the blend. As such the RTD needs to be evaluated over the range of operating condition that can impact process dynamics (e.g. throughput, impeller rotation rate etc.). In this paper, we demonstrate that the dimensionless RTD (normalized with respect to the mean residence time) is invariant with throughput and impeller rotation rates under certain conditions for the two continuous direct compression processes. We present a case study to illustrate the utility of this relationship for predicting the process dynamics at different operating conditions (i.e., throughputs) and evaluating the impact of variations in the process dynamics on the control strategy for a continuous direct compression process.

Given that consumers value quality, and advertising content informs consumers’ beliefs about quality, it is not surprising that high-quality brands emphasize quality in their advertising content. What is less obvious is whether firms with lower-quality brands should also follow suit and emphasize quality in their advertising to signal a higher quality. We examine this issue and study the effectiveness of quality-based advertising messages. Our field study relates brands’ monthly sales to their advertised quality claims across 1,876 print ads in national magazines and Consumer Reports–based product quality ratings over more than two decades. Contrary to the generally held yet erroneous belief in the efficacy of low-quality products emphasizing quality in their advertising, we demonstrate that (1) it is not beneficial for a low-quality firm to emphasize quality in its advertising, and (2) it is effective for a high-quality firm to do so. An analysis of parameter values from a published category-agnostic simulation and an experiment that examines consumers’ responses to quality claims in a second product category yields convergent insights.

Forest and landscape restoration are emerging globally as a major challenge for development and conservation in the twenty-first century. With a restoration market providing demand for participation in supplying products and services, such as native seed, Indigenous communities have experienced new opportunities for cash income and livelihood improvements. This chapter explores the ways different Indigenous populations in Brazil have engaged in the native seed trade for restoring degraded lands, and the outcomes of their participation. We cover a case study based on one decade’s experience of the Xingu Seeds Network, focusing on challenges faced by 232 Indigenous people, mostly women, from 13 villages of Ikpeng, Kawaiwete, Matipu, Panara, Wauja, Xavante and Yudja ethnicity in the southeastern Amazon. Indigenous communities have engaged in the activity mainly to secure current and future access to natural resources. Although communities have shown a large fluctuation in their seed production and cash income over the years, these collectors have produced 6.7 tonnes of seeds from 159 species leading to US$ 65 thousand cash income for households. Native seed production is strongly aligned with traditional knowledge with outcomes related to women’s empowerment, opportunities for young people, territorial mapping, and building local organizations. However, communities have faced constraints in adapting ethnic institutions to business management requirements mainly due to scales of production, language difficulties and lack of accounting skills. Therefore, institutions must be developed acknowledging Indigenous knowledge and culture for building a more inclusive and flexible approach to support Indigenous groups to adapt to participation in markets.

The official approach to urban planning in Thailand involves a centralized process that relies mainly on decisions from the national government. This approach has not been effective in a small and unique area like Wat Ket in Chiang Mai City in northern Thailand which has a variety of different land use types. Some land use areas in the Wat Ket area were designated as commercial zones, without the involvement and agreement of local residents and this led to land use conflicts and other related problems. This paper describes how stakeholders in Wat Ket applied an alternative collaborative approach to urban land use planning. The paper provides a case study of the collaborative urban planning approach in the context of a rapidly growing city in the developing world.Collaborative urban land use planning is supported by land use analysis applying aerial photographs and Geographical Information Systems (GIS) to analyse land use changes in the study area.

Large-scale ecological restoration programs across the world involve a voluminous demand for native seeds of diverse native plant species. In this article, we explore how institutional systems have operated and impacted native seed supply in Brazil. Native seed supply for restoration is essentially a community-based activity which faces broad barriers to operating within regulations because of requirements for excessive and costly technical documentation, scarcity of seed laboratories, and lack of instructions for native seed quality testing. Although decentralized seed networks have stimulated arrangements for local organizations to promote seed supply, policies constrain the development of local capacities and the ongoing sustainability of these organizations. These conditions have resulted in a vast network of informal collectors and producers who are largely “invisible” and unknown to the regulatory authorities. Policies have decentralized responsibilities from the state without devolving decision-making power to the multiple stakeholders engaged in policy elaboration. The policies maintain the centralized regulation of native seed supply. After examining Brazilian seed networks’ experiences and conducting discussions with stakeholders and experts, we suggest adapting the current regulations to more local level contexts, encompassing the following strategies: (1) ensuring native seed origin and identity; (2) relaxation of the laboratory accreditation process for native seed quality assurance; (3) fostering seed markets for restoration; (4) research to provide technological innovation; (5) supporting local, diverse, and small seed-based businesses.

The utility of shell overlays to oyster (Crassostrea virginica) plantings as a cownose ray (Rhinoptera bonasus) predator deterrence mechanism was examined. Typical industry practice of oyster seed planting was followed in an experimental design employing treatment areas of 0.5–1.0 acre (0.2–0.4 hectare). Areas were prepared in the Lower Machodoc Creek, Virginia, by the initial application of shell to insure a stable substrate under planted seed oysters. Seed oysters were planted using standard industry methods. Experimental areas were located, two upstream and two downstream, of a constriction in the Lower Machodoc that dictated differing physical environments in the respective locations with downstream locations being more exposed to northeast wind-driven stresses and, historically, a greater incidence of ray predation. Once oysters were planted, two of the areas, one upstream and one downstream of the aforementioned constriction, were additionally treated with a shell overlay as a predation det...

There is increased research into pharmaceutical nanoemulsions where the timing of quantification of active components can dictate continuous manufacturing production cost and consistency. The goal of this study was development of a rapid quantification method for isoflurane nanoemulsions using Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy (ATR-FTIR). Isoflurane was quantified by ATR-FTIR and High-Performance Liquid Chromatography (HPLC). Correlation and agreement between methods was determined to validate the ATR-FTIR procedure. Evaporation rate studies from the nanoemulsions and pure isoflurane were performed. ATR-FTIR values were in agreement and correlated with (HPLC) measurements (calculated Pearson R of 0.99; 99 % confidence interval; P

Over the last few decades, numerous initiatives have advanced forest landscape restoration in the Amazon, and in 2015 the Brazilian government set an ambitious, still-valid, target to restore 4.8 million hectares of degraded Amazonian land by 2030. This has contributed to an emergent global restoration network that connects multiple stakeholders and processes for funding, implementing and monitoring restoration actions in such a way that prepares various ecosystem services for market integration. The network arose in tandem with the evolution of an institutional framework that includes regulatory requirements within Brazil, global commitments linked to climate change mitigation, corporate sustainability strategies, and the growth of crowd-sourcing activism. This paper presents restoration activities as embedded within a Global Production Network (GPN) for an ecosystem service, which we use as a heuristic device to inform our understanding of emergent environmental governance structures. The resulting multi-scalar, networked mode of environmental governance is presented as a web-like structure co-created by institutional evolution, actor-specific strategies, and interactions between firms and non-firm actors. The article pays particular attention to a case study of how the restoration network manifests territorially in the Upper Xingu region of the Brazilian Amazon. Despite the strong North-South orientation of dominant funding relationships, network governance is also seen to be relational. This is evident from the dissemination of ideas, supply models and seeding techniques from Upper Xingu to other regions of Brazil. These insights could be applied to improve landscape restoration outcomes, and indeed the provisioning of ecosystem services more broadly.

In this paper, we explore how diverse community networks in Brazil have locally advanced seed production and institutional systems to enhance a restoration economy. By focusing on the experiences of the six major native seed suppliers in Amazonia, the Cerrado, and the Atlantic Forest, we estimate the capacity to scale-up community-based systems to meet a large-scale restoration target as a rural development strategy. Over one decade, 1016 collectors traded 416.91 tonnes of native seeds representing, on average, 31.41 kilos yearly and USD 256.5 as household income. Based on this well documented empirical evidence, we estimate that Brazil&rsquo, s restoration goal would require from 3.6 to 15.6 thousand tonnes of native seeds depending on the share of each restoration method adopted with potential work opportunities for 13.2 to 57.1 thousand collectors yearly and total income from USD 34 to 146 million. We argue that community networks represent feasible arrangements for increasing the availability of plant material sources which provide high socio-economic benefits. For scaling up native seed sources, we suggest the following key strategies: (i) government incentives and subsidies, (ii) enforcement of ecosystem restoration, (iii) community participation, (iv) adaptation of the seed regulations, (v) technological development, and (vi) seed market diversification.

While community forestry has shown promise to reduce rural poverty, improve reforestation and potentially offset carbon emissions, many projects have failed, either partly or completely. In order to understand why community forestry succeeds or fails, we examined in detail the literature related to community forestry from three countries, Mexico, Nepal and the Philippines. We also drew on experiences in other countries in Asia, Latin America and Africa. We identified five main interconnected factors which the literature suggests are often critical to the success of community forestry. To integrate the many ways in which community forestry projects can improve the state of these factors, we use the concept of ‘bonding social capital’, i.e. communities’ ability to work together towards a common aim and ‘bridging social capital’, i.e. their ability to liaise with the outside world. To understand the interaction of the five success factors and the way in which improvements to bonding or bridging social capital may affect them, we developed a causal diagram which depicts the interrelationships between the success factors and the key points at which project inputs may be best applied. It is clear from our analysis that failing to appreciate both the complexity and interaction of the various influences may lead to project failure.

Community forestry focuses on the link between forest resources and livelihoods and contributes to forest conservation and reforestation. It is widespread in Nepal, with a very high proportion of the rural population involved, and is widely recognized as one of the most successful examples of community forestry in Asia. Through a combination of literature reviews and original research, this volume explores key experiences and outcomes of community forestry in Nepal over the last four decades as a model for improving forest management and supporting local livelihoods. The book takes a critical approach, recognizing successes, especially in forest conservation and restoration, along with mixed outcomes in terms of poverty reduction and benefits to forest users. It recognizes the way that community forestry has continued to evolve to meet new challenges, including the global challenges of climate change, environmental degradation and conservation, as well as national demographic and social changes due to large-scale labour migration and the growing remittance economy. In addition to examining the changes and responses, the book explores ways that community forestry in Nepal might move forward. Lessons from Nepal have relevance to community forestry and community-based approaches to natural resource management around the world that are also experiencing global pressures and opportunities. [Book Synopsis]

Community forestry focuses on the link between forest resources and livelihoods and contributes to forest conservation and reforestation. It is widespread in Nepal, with a very high proportion of the rural population involved, and is widely recognized as one of the most successful examples of community forestry in Asia. Through a combination of literature reviews and original research, this volume explores key experiences and outcomes of community forestry in Nepal over the last four decades as a model for improving forest management and supporting local livelihoods. The book takes a critical approach, recognizing successes, especially in forest conservation and restoration, along with mixed outcomes in terms of poverty reduction and benefits to forest users. It recognizes the way that community forestry has continued to evolve to meet new challenges, including the global challenges of climate change, environmental degradation and conservation, as well as national demographic and social changes due to large-scale labour migration and the growing remittance economy. In addition to examining the changes and responses, the book explores ways that community forestry in Nepal might move forward. Lessons from Nepal have relevance to community forestry and community-based approaches to natural resource management around the world that are also experiencing global pressures and opportunities. [Book Synopsis]

Cell motility is required for diverse processes during immunity and inflammation. Classically, leukocyte motility is defined as an amoeboid type of migration, however some leukocytes, like macrophages, also employ a more mesenchymal mode of migration. Here, we sought to characterize the mechanisms that regulate neutrophil and macrophage migration in vivo by using real-time imaging of leukocyte motility within interstitial tissues in zebrafish larvae. Neutrophils displayed a rounded morphology and rapid protease-independent motility, lacked defined paxillin puncta, and had persistent rearward polarization of stable F-actin and the microtubule network. By contrast, macrophages displayed an elongated morphology with reduced speed and increased directional persistence and formed paxillin-containing puncta but had a less-defined polarization of the microtubule and actin networks. We also observed differential effects of protease inhibition, microtubule disruption and ROCK inhibition on the efficiency of neutrophil and macrophage motility. Taken together, our findings suggest that larval zebrafish neutrophils and macrophage display distinct modes of migration within interstitial tissues in vivo .

Parents and other adults such as baby sitters, daycare workers, and teachers are responsible for making choices that contribute to the development and well-being of the children in their care. In a typical day, for example, parents decide what and how much their children eat, the amount of time they spend online, the television programs they watch, and the time they go to bed. In this research, we identify two common approaches -imposition and accommodation-that are available to decision makers and outline how these decisions affect the feelings of power and eating behaviors of the adults themselves. Four experiments and one field study demonstrate that adults shape their own consumption choices based on whether they accommodate the virtuous (i.e. healthy) vs. indulgent (i.e. unhealthy) preferences of the child or impose their own preferences. We find that when adults accommodate the child's preferences, they choose healthier foods for themselves. Finally, we demonstrate that the social context of consumption moderates these effects. We find support for our power-based theory in five studies using a variety of methods, participants, and contexts. The results have important implications for the health of adults who take care of children.

One of the key elements in modern reservoir exploration and management is describing reservoir fluid phase behavior and physical properties commonly referred to as pressure-volume-temperature, or PVT data. Typically, PVT data come from laboratory tests, empirical correlations, and Equation of State (EOS) models. It is common practice to describe the phase behavior or PVT data through EOS models tuned to laboratory measurements on reservoir fluid samples. After a sample is received at a laboratory, a portfolio of PVT laboratory tests are performed. The results are quality checked and the appropriate data are selected to tune an EOS model to obtain an accurate EOS description of the reservoir fluid. Each step in this process requires judgment and decisions from the corresponding domain experts to achieve physically sound PVT relations and calculate the required properties. Such EOS based modelling processes are time consuming, expensive and exposed to various risks due to multiple human interventions. In the work presented in this paper, a study was conducted to explore the feasibility of a defined step EOS based modelling workflow using a limited, but defined, laboratory data set as the basis for characterizing and tuning the EOS model. In the development of a standardized workflow, a variety of EOS characterization and tuning methods were established to accommodate the diverse and complex nature of reservoir fluids. The methods considered include a modified Pedersen's method, a gamma distribution based method, and two methods based on single carbon number (SCN) composition and aromaticity factors. Despite their differences, the methods follow the principal objectives to be operator independent, robust, thermodynamically consistent, and numerically simple. Apart from the fluid composition, the only PVT data required for the proposed workflow were the saturation pressure, densities and the volumetric data obtained from constant composition expansion (CCE) measurements. The CCE test is advantageous because it is non-destructive to the sample being tested and can be performed quickly and reliably either in the laboratory or at a well-site. An optimized EOS model utilizing appropriate fluid characterization and tuning method is then selected based on a pre-defined Key Performance Indicator (KPI) derived from the deviation of the model predictions with the experimental data. With this optimal model defined, all other PVT data, such as those from sample destructive differential liberation (DL) or constant volume depletion (CVD) tests, can be reliably predicted. The workflow was validated using extensive PVT data measured for a variety of reservoir fluids including black oils, volatile oils, and gas condensates. The results have shown that the proposed workflow can reliably model PVT phase behavior and properties for the majority of tested reservoir fluids. This article will thoroughly discuss the details of this workflow and the modelling results on four benchmark fluids of different types.