Aptamers as affinity ligands for downstream processing

Aptamers are single‐stranded synthetic oligonucleotides that are able to capture their target molecule with high affinity and specificity. Therefore, they can be thought of as nucleic acid‐based alternative to antibodies, which have several advantages over their amino acid‐based counterparts. Consequently, aptamers can be used in different applications based on molecular recognition including affinity separations. This review will summarize the state‐of‐the‐art in aptamer‐based affinity separations; will discuss the current limitations and will highlight possible future prospects. The first part will point out the advantages of aptamers in downstream processing. Here, the properties of aptamers will be discussed along with their implications on downstream processing from a user's point of view. In the second part, a brief summary of the literature is given with focus on aptamer‐based separation of proteins. Finally, some drawbacks of aptamers will be illustrated and possibilities to overcome these limitations will be suggested. New technologies in the fields of aptamer selection and synthesis are expected to further promote the use of aptamers as affinity ligands in downstream processing.     

Isolation and characterization of cell membranes

The steps outlined above should provide a framework for developing and evaluating a scheme for isolating a particular membrane system of interest. Further characterization often focuses on the specific research interests of an investigator. Most subsequent techniques will probably involve biochemical analyses of protein and lipid components and biophysical analyses of membrane properties. The techniques to be used, such as SDS gel electrophoresis, enzyme kinetics, receptor binding, protein purification, etc. are not unique to membrane systems and will not be described in detail. However, it is likely that the functional relevance and success of subsequent analyses will be aided if the starting membranes are as homogeneous and well-characterized as possible. Even if some of the fractionation steps are bypassed, such as might occur if a specific membrane protein is detergent-extracted, purified, and reconstituted directly from a crude homogenate, a full understanding of membrane protein function will probably require complementary studies on intact cells and isolated membranes.     

Development of FuGO: an ontology for functional genomics investigations

The development of the Functional Genomics Investigation Ontology (FuGO) is a collaborative, international effort that will provide a resource for annotating functional genomics investigations, including the study design, protocols and instrumentation used, the data generated and the types of analysis performed on the data. FuGO will contain both terms that are universal to all functional genomics investigations and those that are domain specific. In this way, the ontology will serve as the "semantic glue" to provide a common understanding of data from across these disparate data sources. In addition, FuGO will reference out to existing mature ontologies to avoid the need to duplicate these resources, and will do so in such a way as to enable their ease of use in annotation. This project is in the early stages of development; the paper will describe efforts to initiate the project, the scope and organization of the project, the work accomplished to date, and the challenges encountered, as well as future plans.     

Climate change and health with an emphasis on interactions with ultraviolet radiation: a review

Climate change is increasingly recognized as a major risk to human health, and health concerns are assuming more importance in international debates on mitigation and adaptation strategies. Health consequences of climate change will occur through direct and indirect routes, and as a result of interactions with other environmental exposures. Heatwaves will become more common and are associated with higher mortality particularly in the elderly and those with pre‐existing cardiovascular and respiratory illnesses. Warmer ambient temperatures will result in more dehydration episodes and increased risks of renal disease and, through effects on pollen seasons, there may be an increase in allergic disease such as asthma and hayfever. Other adverse effects including on air quality, food safety and security and an expanding distribution of some infectious diseases, including vector‐borne diseases, are postulated. A related but separate environmental exposure is that of ultraviolet radiation (UVR). Interactions between climate change and stratospheric ozone (and the causes of ozone depletion) will cause changes to levels of ambient UVR in the future and warmer temperatures are likely to change sun exposure behaviour. Co‐occurring effects on aquatic and terrestrial ecosystems have potential consequences for food safety, quality and supply. Climate change‐related exposures are likely to affect the incidence and distribution of diseases usually considered as caused by UVR exposure; and changes in UVR exposure will modulate the climate change effects on human health. For example, in some regions warmer temperatures due to climate change will encourage more outdoor behaviour, with likely consequences for increasing skin cancer incidence. Although many of the health outcomes of both climate change and the interaction of climate change and UVR exposure are somewhat speculative, there are risks to over‐ or under‐estimations of health risks if synergistic and antagonistic effects of co‐occurring environmental changes are not considered.     

Alteration of the phenology of leaf senescence and fall in winter deciduous species by climate change: effects on nutrient proficiency

Leaf senescence in winter deciduous species signals the transition from the active to the dormant stage. The purpose of leaf senescence is the recovery of nutrients before the leaves fall. Photoperiod and temperature are the main cues controlling leaf senescence in winter deciduous species, with water stress imposing an additional influence. Photoperiod exerts a strict control on leaf senescence at latitudes where winters are severe and temperature gains importance in the regulation as winters become less severe. On average, climatic warming will delay and drought will advance leaf senescence, but at varying degrees depending on the species. Warming and drought thus have opposite effects on the phenology of leaf senescence, and the impact of climate change will therefore depend on the relative importance of each factor in specific regions. Warming is not expected to have a strong impact on nutrient proficiency although a slower speed of leaf senescence induced by warming could facilitate a more efficient nutrient resorption. Nutrient resorption is less efficient when the leaves senesce prematurely as a consequence of water stress. The overall effects of climate change on nutrient resorption will depend on the contrasting effects of warming and drought. Changes in nutrient resorption and proficiency will impact production in the following year, at least in early spring, because the construction of new foliage relies almost exclusively on nutrients resorbed from foliage during the preceding leaf fall. Changes in the phenology of leaf senescence will thus impact carbon uptake, but also ecosystem nutrient cycling, especially if the changes are consequence of water stress.     

Histone chaperones, a supporting role in the limelight

In eukaryotic cells, highly basic histone proteins are associated with the DNA to form the nucleosome, the fundamental unit of chromatin. Histones are closely escorted by histone chaperones from their point of synthesis up to their delivery site. We will present an overview of the histone chaperones identified to date with their various roles, in an attempt to highlight their importance in cellular metabolism. Nucleoplasmin will illustrate a role in histone storage and Nap-1, a histone translocator. CAF-1 and Hira will provide examples of distinct histone deposition factors coupled to and uncoupled from DNA synthesis, respectively, while Asf1 could act as a histone donor. We then will illustrate with two examples how histone chaperones can be associated with chromatin remodeling activities. Finally, we will discuss how the RbAp46/48 proteins, as escort factors, are part of multiple complexes with various functions. Based on these examples, we will propose a scheme in which the diverse roles of histone chaperones are integrated within an assembly line for chromatin formation and regulation. Finally, we discuss how these chaperones may have more than a supporting role in a histone metabolic pathway.     

Will the "new biology" affect the future of histopathology?

The technologies used in histopathology are changing as a consequence of the current revolutionary progress in several areas of biology. It is likely that general cancer management will improve because of the impact of molecular techniques and immunohistochemistry on tumor diagnosis and classification and on the determination of prognosis and response to therapy. Moreover, as therapies are starting to be modelled after the distinctive molecular characteristics of a specific tumor, the availability of molecular tests to all patients will become a matter of great importance.     

The essential characteristics of a rating system

The principles of the Tanner-Whitehouse-Healy system for assessing skeletal maturity will be described and in particular the statistical basis for the scores given to each stage of each bone in the hand and wrist. The uses of the system, particularly in clinical cases, will be discussed together with reliability of the ratings. A revised system will be presented in which a part of the stages are amalgamated and the possibility of dropping some of the bones will be discussed. General discussion will be invited on the logical basis for selecting certain stages of certain bones as superior to others as assessors of maturity.     

胞外体--免疫治疗中的"特洛伊木马"

胞外体是源于多种真核细胞的多泡体,通过后者与质膜融合释放到细胞外的一种膜性小囊泡,在多种生理过程中发挥作用。近年研究发现,由抗原提呈细胞分泌的胞外体富集MHCI/II类分子、协同刺激分子、热休克蛋白70和热休克蛋白90等多种生物活性分子于一身,像“特洛伊木马”一样,在体内外免疫调节中起关键作用。本文就胞外体的基本特征、生产纯化方法及其作为一种新型的亚细胞疫苗在抗肿瘤和抗病毒免疫中的应用前景予以综述。     

Abiotic Resource Depletion Different perceptions of the problem with mineral deposits

Goal, Scope and Background The goal of the present paper is to demonstrate how environmental product declarations (EPDs) are developed based on a set of product category rules (PCRs) in accordance with the requirements in the ISO 14025-standard. This is demonstrated by examples from the furniture industry in Norway, where several case models are evaluated. To ease the capability of developing EPDs in this industry, a database with specific environmental data for materials in furniture is developed. The database is used to produce the LCA for selected furniture models, and further, the database is the backbone of a data-assistance tool used to create the EPDs. Methods The LCA-data are produced based on traditional LCA-methodology. The PCR is based on a stakeholder analysis and the proposed methodology in the ISO 14025-standard. The EPDs developed so far, are results of close collaboration between companies and research centres in the Nordic countries. For the verification of the EPDs, auditing methodologies are used as a part of the audit of the companies' environmental management systems (EMS). Results and Conclusion Based on a hearing of a set of suggested PCRs, a consensus document for seating accommodation is developed. This is further the model for how to develop PCR-documents for all types of furniture, for example sleeping accommodations. Likewise, the database shall contain the most important data for the parts of a furniture model. Within the goal of the project period, EPDs will be developed for 80% of Norwegian furniture. The verification of the EPDs is done as a part of the certification procedures of EMS in accordance with the ISO 14001. Recommendation and Perspective The results presented in the paper are mainly for the pilot models in the project. However, the results will be further tested and the data-tool will be developed as a part of a product design tool where environmental requirements will be combined with quality requirements. The product design tool will be implemented in the furniture industry. Information on how to use EPDs in public purchasing will also be a part of future work.     

Editorial: Writing in Grana

It has been necessary to re-organize the production of GRANA. The following changes in publishing policy are the most important. 1. GRANA will continue to accept papers in its traditional field of interest, but in future, papers in the applied branches of Palynology, such as pollen-analytical studies, will also be published, as well as papers on other micropalae-ontological fields, in which the methods correspond to those used in palynological research (dinoflagellates, chitinozoa, diatoms, etc.). Only material that is obviously of international interest will be accepted. 2. From now on, there will be restrictions as to the length of a paper. Two types of contributions will be acceptable: Articles, containing 16 printed pages at the most, including the figures, and Notes, covering four printed pages at the most. The Notes will be given priority over the Articles, and will be included in the journal as soon as they have been scrutinized by a referee and prepared for publication by the editorial staff. Thus, the writing of Notes will be the quick way to secure publication in GRANA. The rest of this article gives the technical specifications of GRANA and also some instructions for authors.     

Biopolymer-based hydrogels as scaffolds for tissue engineering applications: a review

Hydrogels are physically or chemically cross-linked polymer networks that are able to absorb large amounts of water. They can be classified into different categories depending on various parameters including the preparation method, the charge, and the mechanical and structural characteristics. The present review aims to give an overview of hydrogels based on natural polymers and their various applications in the field of tissue engineering. In a first part, relevant parameters describing different hydrogel properties and the strategies applied to finetune these characteristics will be described. In a second part, an important class of biopolymers that possess thermosensitive properties (UCST or LCST behavior) will be discussed. Another part of the review will be devoted to the application of cryogels. Finally, the most relevant biopolymer-based hydrogel systems, the different methods of preparation, as well as an in depth overview of the applications in the field of tissue engineering will be given.     

Release of plant-borne flavonoids into the rhizosphere and their role in plant nutrition

Plants release a multitude of organic compounds into the rhizosphere, some of which are flavonoids. These products of secondary metabolism are mainly studied for their antioxidant properties and for their role in the establishment of rhizobium-legume symbiosis; however, it has been recently demonstrated that flavonoids can also affect nutrient availability through soil chemical changes. This review will give an overview of the types and amounts of flavonoids released by roots of different plant species, as well as summarize the available knowledge on root exudation mechanisms. Subsequently, factors influencing their release will be reported, and the methodological approaches used in the literature will be critically described. Finally, the direct contribution of plant-borne flavonoids on the nitrogen, phosphorous and iron availability into the rhizosphere will be discussed.     

Rotating wave solutions of the FitzHugh–Nagumo equations

This paper will treat the bifurcation and numerical simulation of rotating wave (RW) solutions of the FitzHugh-Nagumo (FHN) equations. These equations are often used as a simple mathematical model of excitable media. The dependence of the solutions on a uniformly applied current, and also on the diffusion coefficients or domain size will be studied. Ranges of applied current and/or diffusion coefficients in which RW solutions are observed will be described using bifurcation theory and continuation methods. The bifurcation of time-periodic solutions of these FHN equations without diffusion is described first. Similar methods are then used to find RW solutions on a circular ring and numerical simulations are described. These results are then extended to investigate RW solutions on annular rings of finite cross-section. Scaling arguments are used to show how the existence of solutions depends on either the diffusion coefficient or on the size of the region.     

Rational Vector Design for Efficient Non-viral Gene Delivery: Challenges Facing the Use of Plasmid DNA

Although non-viral gene delivery is a very straightforward technology, there are currently no FDA-approved gene medicinal products available. Therefore, improving potency, safety, and efficiency of current plasmid DNA vectors will be a major task for the near future. This article will provide an overview on factors influencing production yield and quality as well as safety issues that emerge from the vector design itself. Special focus will be on generating bacterial pDNA vectors by circumventing the use of antibiotic resistance genes, to generate safer gene medicinal products as well as smaller, more efficient DNA vectors.     

Brazil and the development of international scientific biosafety testing guidelines for transgenic crops

Under the umbrella of the International Organisation of Biological Control (IOBC), an international working group of public sector scientists entitled on "Transgenic Organisms in Integrated Pest Management and Biological Control" has been organized. The group will develop scientific principles and detailed scientific guidelines for biosafety testing of transgenic crops. The key elements of this project are: (1) An international initiative including expert scientists from leading research institutions in developed and developing countries; (2) coordination of the development and implementation of the guidelines as a dynamic process, which will include scientific and technical capacity building and communication among scientists and between scientists and policy makers; (3) rapid serial publication of sections of the guidelines as they are completed; and (4) rapid and timely revision of previously published sections. The guidelines will be constructed on a case-by-case basis and will have no regulatory legitimacy themselves.     

Incorporation of macromolecules in microparticles: preparation and characteristics.

Macromolecules can easily be incorporated in microparticles of polyacrylamide by copolymerization with acrylamide in a water-in-oil emulsion. The microparticles preferably arount 1-3 mum in diameter, will have a macroporous structure formed by the polymeric network. The amount of incorporation of the macromolecules will depend on the structure of the network, which, in turn, will depend on the total amount of monomer (T) and the relative amount of cross-linking agent (C) in the monomeric solution. Two mechanisms are responsible for the incorporation; all macromolecules are, independently of the size, fixed in the threads of polyacrylamide and large ones are entrapped within the network formed by the threads. The amount entrapped will depend on the size of the macromolecule and the mean pore radius of the gel. In microparticles with a total concentration of monomers of 8% and a cross-linking of 25% (T-C=8-25) the biological properties of incorporated macromolecules are retained, due to the macroporous structure, as found in binding studies with albumin. The density of the particles will also depend on C and T and, to some extent, on the protein concentration. Due to the fixation in the polyacrylamide threads, some of the incorporated macromolecules will be exposed on the surface, allowing them to react with, for instance, cells, which cannot penetrate the particles. The optimal conditions for the incorporation of macromolecules in the microparticles are investigated.     

Cell surface heparan sulfate proteoglycan and the neoplastic phenotype

Cell surface proteoglycans are strategically positioned to regulate interactions between cells and their surrounding environment. Such interactions play key roles in several biological processes, such as cell recognition, adhesion, migration, and growth. These biological functions are in turn necessary for the maintenance of differentiated phenotype and for normal and neoplastic development. There is ample evidence that a special type of proteoglycan bearing heparan sulfate side chains is localized at the cell surface in a variety of epithelial and mesenchymal cells. This molecule exhibits selective patterns of reactivity with various constituents of the extracellular matrix and plasma membrane, and can act as growth modulator or as a receptor. Certainly, during cell division, membrane constituents undergo profound rearrangement, and proteoglycans may be intimately involved in such processes. The present work will focus on recent advances in our understanding of these complex macromolecules and will attempt to elucidate the biosynthesis, the structural diversity, the modes of cell surface association, and the turnover of heparan sulfate proteoglycans in various cell systems. It will then review the multiple proposed roles of this molecule, with particular emphasis on the binding properties and the interactions with various intracellular and extracellular elements. Finally, it will focus on the alterations associated with the neoplastic phenotype and will discuss the possible consequences that heparan sulfate may have on the growth of normal and transformed cells.