Sucrose density gradient centrifugation and cross-flow filtration methods for the production of arbovirus antigens inactivated by binary ethylenimine

BACKGROUND: Sucrose density gradient centrifugation and cross-flow filtration methods have been developed and standardised for the safe and reproducible production of inactivated arbovirus antigens which are appropriate for use in diagnostic serological applications. METHODS: To optimise the maximum titre of growth during the propagation of arboviruses, the multiplicity of infection and choice of cell line were investigated using stocks of Ross River virus and Barmah Forest virus grown in both mosquito and mammalian cell lines. To standardise and improve the efficacy of the inactivation of arboviral suspensions, stocks of Ross River virus, Barmah Forest virus, Japanese encephalitis virus, Murray Valley encephalitis virus and Alfuy virus were chemically inactivated using binary ethylenimine at a final concentration of 3 mM. Aliquots were then taken at hourly intervals and crude inactivation rates were determined for each virus using a plaque assay. To ensure complete inactivation, the same aliquots were each passaged 3 times in Aedes albopictus C6/36 cells and the presence of viral growth was detected using an immunofluorescent assay. For larger quantities of viral suspensions, centrifugation on an isopycnic sucrose density gradient or cross-flow filtration was used to produce concentrated, pure antigens or partially concentrated, semi-purified antigens respectively. RESULTS: The results of the propagation experiments suggested that the maximum viral titres obtained for both Ross River virus and Barmah Forest virus were affected by the incubation period and choice of cell line, rather than the use of different multiplicity of infection values. Results of the binary ethylenimine inactivation trial suggested that standardised periods of 5 or 8 hours would be suitable to ensure effective and complete inactivation for a number of different arboviral antigens. CONCLUSION: Two methods used to prepare inactivated arbovirus antigens have been standardised to minimise production failure and expenditure and to provide reagents that conform to the highest quality and safety requirements of a diagnostic serology laboratory. The antigens are suitable for use in either enzyme linked immunosorbent assays or haemagglutination inhibition assays and the optimised protocols can be directly applied to produce antigens from new or emerging arboviral pathogens

Range-wide assessment of livestock grazing across the sagebrush biome

Domestic livestock grazing occurs in virtually all sagebrush habitats and is a prominent disturbance factor. By affecting habitat condition and trend, grazing influences the resources required by, and thus, the distribution and abundance of sagebrush-obligate wildlife species (for example, sage-grouse Centrocercus spp.). Yet, the risks that livestock grazing may pose to these species and their habitats are not always clear. Although livestock grazing intensity and associated habitat condition may be known in many places at the local level, we have not yet been able to answer questions about use, condition, and trend at the landscape scale or at the range-wide scale for wildlife species. A great deal of information about grazing use, management regimes, and ecological condition exists at the local level (for individual livestock management units) under the oversight of organizations such as the Bureau of Land Management (BLM). However, the extent, quality, and types of existing data are unknown, which hinders the compilation, mapping, or analysis of these data. Once compiled, these data may be helpful for drawing conclusions about rangeland status, and we may be able to identify relationships between those data and wildlife habitat at the landscape scale

A Bose-Einstein Approach to the Random Partitioning of an Integer

Consider N equally-spaced points on a circle of circumference N. Choose at random n points out of $N$ on this circle and append clockwise an arc of integral length k to each such point. The resulting random set is made of a random number of connected components. Questions such as the evaluation of the probability of random covering and parking configurations, number and length of the gaps are addressed. They are the discrete versions of similar problems raised in the continuum. For each value of k, asymptotic results are presented when n,N both go to infinity according to two different regimes. This model may equivalently be viewed as a random partitioning problem of N items into n recipients. A grand-canonical balls in boxes approach is also supplied, giving some insight into the multiplicities of the box filling amounts or spacings. The latter model is a k-nearest neighbor random graph with N vertices and kn edges. We shall also briefly consider the covering problem in the context of a random graph model with N vertices and n (out-degree 1) edges whose endpoints are no more bound to be neighbors

Transcriptional regulation of the urokinase receptor (u-PAR) - A central molecule of invasion and metastasis

The phenomenon of tumor-associated proteolysis has been acknowledged as a decisive step in the progression of cancer. This short review focuses on the urokinase receptor (u-PAR), a central molecule involved in tumor-associated invasion and metastasis, and summarizes the transcriptional regulation of u-PAR. The urokinase receptor (u-PAR) is a heavily glycosylated cell surface protein and binds the serine protease urokinase specifically and with high affinity. It consists of three similar cysteine-rich repeats and is anchored to the cell membrane via a GPI-anchor. The u-PAR gene comprises 7 exons and is located on chromosome 19q13. Transcriptional activation of the u-PAR promoter region can be induced by binding of transcription factors (Sp1, AP-1, AP-2, NF-kappaB). One current study gives an example for transcriptional downregulation of u-PAR through a PEA3/ets transcriptional silencing element. Knowledge of the molecular regulation of this molecule in tumor cells could be very important for diagnosis and therapy in the near future

From evidence to action: applying gender mainstreaming to pay gaps in the Welsh public sector

Progress on reducing gender disparities remains painfully slow, despite efforts to identify the determinants of gender pay gaps and specify size and shape. Recent studies highlight the need for a more nuanced account of the way that public policy shapes organizational responses and insights into the types of organizational practices that diminish pay disparities. In response, this research reports on an action research intervention in three large Welsh public organizations, subject to a unique statutory equality duty. Data demonstrate how an evidence‐based gender mainstreaming approach facilitated the development of a ‘no blame’ strategy, which legitimized organizational proactivity through collaborative and empowering change management processes. The research contributes to the study of gender pay gaps by demonstrating that gender mainstreaming, with facilitative local conditions and supportive public policy, shapes action on gender segregation, with particular success in women's low‐paid employment. Conclusions highlight theoretical and policy implications arising from the research

MASTR-MS: A web-based collaborative laboratory information management system (LIMS) for metabolomics

Background An increasing number of research laboratories and core analytical facilities around the world are developing high throughput metabolomic analytical and data processing pipelines that are capable of handling hundreds to thousands of individual samples per year, often over multiple projects, collaborations and sample types. At present, there are no Laboratory Information Management Systems (LIMS) that are specifically tailored for metabolomics laboratories that are capable of tracking samples and associated metadata from the beginning to the end of an experiment, including data processing and archiving, and which are also suitable for use in large institutional core facilities or multi-laboratory consortia as well as single laboratory environments. Results Here we present MASTR-MS, a downloadable and installable LIMS solution that can be deployed either within a single laboratory or used to link workflows across a multisite network. It comprises a Node Management System that can be used to link and manage projects across one or multiple collaborating laboratories; a User Management System which defines different user groups and privileges of users; a Quote Management System where client quotes are managed; a Project Management System in which metadata is stored and all aspects of project management, including experimental setup, sample tracking and instrument analysis, are defined, and a Data Management System that allows the automatic capture and storage of raw and processed data from the analytical instruments to the LIMS. Conclusion MASTR-MS is a comprehensive LIMS solution specifically designed for metabolomics. It captures the entire lifecycle of a sample starting from project and experiment design to sample analysis, data capture and storage. It acts as an electronic notebook, facilitating project management within a single laboratory or a multi-node collaborative environment. This software is being developed in close consultation with members of the metabolomics research community

Restoration Handbook for Sagebrush Steppe Ecosystems with Emphasis on Greater Sage-Grouse Habitat—Part 3. Site Level Restoration Decisions

Sagebrush steppe ecosystems in the United States currently (2016) occur on only about one-half of their historical land area because of changes in land use, urban growth, and degradation of land, including invasions of non-native plants. The existence of many animal species depends on the existence of sagebrush steppe habitat. The greater sage-grouse (Centrocercus urophasianus) depends on large landscapes of intact habitat of sagebrush and perennial grasses for their existence. In addition, other sagebrush-obligate animals have similar requirements and restoration of landscapes for greater sage-grouse also will benefit these animals. Once sagebrush lands are degraded, they may require restoration actions to make those lands viable habitat for supporting sagebrush-obligate animals, livestock, and wild horses, and to provide ecosystem services for humans now and for future generations. When a decision is made on where restoration treatments should be applied, there are a number of site-specific decisions managers face before selecting the appropriate type of restoration. This site-level decision tool for restoration of sagebrush steppe ecosystems is organized in nine steps. ●Step 1 describes the process of defining site-level restoration objectives. ●Step 2 describes the ecological site characteristics of the restoration site. This covers soil chemistry and texture, soil moisture and temperature regimes, and the vegetation communities the site is capable of supporting. ●Step 3 compares the current vegetation to the plant communities associated with the site State and Transition models. ●Step 4 takes the manager through the process of current land uses and past disturbances that may influence restoration success. ●Step 5 is a brief discussion of how weather before and after treatments may impact restoration success. ●Step 6 addresses restoration treatment types and their potential positive and negative impacts on the ecosystem and on habitats, especially for greater sage-grouse. We discuss when passive restoration options may be sufficient and when active restoration may be necessary to achieve restoration objectives. ●Step 7 addresses decisions regarding post-restoration livestock grazing management. ●Step 8 addresses monitoring of the restoration; we discuss important aspects associated with implementation monitoring as well as effectiveness monitoring. ●Step 9 takes the information learned from monitoring to determine how restoration actions in the future might be adapted to improve restoration success

Restoration Handbook for Sagebrush Steppe Ecosystems with Emphasis on Greater Sage-Grouse Habitat—Part 2. Landscape Level Restoration Decisions

Sagebrush steppe ecosystems in the United States currently (2015) occur on only about one-half of their historical land area because of changes in land use, urban growth, and degradation of land, including invasions of non-native plants. The existence of many animal species depends on the existence of sagebrush steppe habitat. The greater sage-grouse (Centrocercus urophasianus) is a landscape-dependent bird that requires intact habitat and combinations of sagebrush and perennial grasses to exist. In addition, other sagebrush-obligate animals also have similar requirements and restoration of landscapes for greater sage-grouse also will benefit these animals. Once sagebrush lands are degraded, they may require restoration actions to make those lands viable habitat for supporting sagebrush-obligate animals. Land managers do not have resources to restore all locations because of the extent of the restoration need and because some land uses are not likely to change, therefore, restoration decisions made at the landscape to regional scale may improve the effectiveness of restoration to achieve landscape and local restoration objectives. We present a landscape restoration decision tool intended to assist decision makers in determining landscape objectives, to identify and prioritize landscape areas where sites for priority restoration projects might be located, and to aid in ultimately selecting restoration sites guided by criteria used to define the landscape objectives. The landscape restoration decision tool is structured in five sections that should be addressed sequentially. Each section has a primary question or statement followed by related questions and statements to assist the user in addressing the primary question or statement. This handbook will guide decision makers through the important process steps of identifying appropriate questions, gathering appropriate data, developing landscape objectives, and prioritizing landscape patches where potential sites for restoration projects may be located. Once potential sites are selected, land managers can move to the site-specific decision tool to guide restoration decisions at the site level

Restoration Handbook for Sagebrush Steppe Ecosystems with Emphasis on Greater Sage-Grouse Habitat—Part 1. Concepts for Understanding and Applying Restoration

Sagebrush steppe ecosystems in the United States currently occur on only about one-half of their historical land area because of changes in land use, urban growth, and degradation of land, including invasions of non-native plants. The existence of many animal species depends on the existence of sagebrush steppe habitat. The greater sage-grouse (Centrocercus urophasianus) is a landscape-dependent bird that requires intact habitat and combinations of sagebrush and perennial grasses to exist. In addition, other sagebrush-obligate animals also have similar requirements and restoration of landscapes for greater sage-grouse also will benefit these animals. Once sagebrush lands are degraded, they may require restoration actions to make those lands viable habitat for supporting sagebrush-obligate animals. This restoration handbook is the first in a three-part series on restoration of sagebrush ecosystems. In Part 1, we discuss concepts surrounding landscape and restoration ecology of sagebrush ecosystems and greater sage-grouse that habitat managers and restoration practitioners need to know to make informed decisions regarding where and how to restore specific areas. We will describe the plant dynamics of sagebrush steppe ecosystems and their responses to major disturbances, fire, and defoliation. We will introduce the concepts of ecosystem resilience to disturbances and resistance to invasions of annual grasses within sagebrush steppe. An introduction to soils and ecological site information will provide insights into the specific plants that can be restored in a location. Soil temperature and moisture regimes are described as a tool for determining resilience and resistance and the potential for various restoration actions. Greater sage-grouse are considered landscape birds that require large areas of intact sagebrush steppe; therefore, we describe concepts of landscape ecology that aid our decisions regarding habitat restoration. We provide a brief overview of restoration techniques for sage-grouse habitat restoration. We conclude with a description of the critical nature of monitoring for adaptive management of sagebrush steppe restoration at landscape- and project-specific levels

Long and short paths in uniform random recursive dags

In a uniform random recursive k-dag, there is a root, 0, and each node in turn, from 1 to n, chooses k uniform random parents from among the nodes of smaller index. If S_n is the shortest path distance from node n to the root, then we determine the constant \sigma such that S_n/log(n) tends to \sigma in probability as n tends to infinity. We also show that max_{1 \le i \le n} S_i/log(n) tends to \sigma in probability.Comment: 16 page