Fuzzy methods for assessing criteria and indicators of sustainable forest management

This paper describes the general concepts, meaning, and definitions of sustainability and proposes the use of soft methodologies, particularly fuzzy set theory, for its assessment. Criteria and indicators (C&I) are described as instruments to assess forest sustainability. Basic elements and concepts of fuzzy sets are described, including membership functions and their interpretations in the context of sustainable forest management. Moreover, fuzzy operators that can combine the operational concepts of sustainability, namely criteria and indicators are described. A simple illustrative example is described to demonstrate the application of these methodologies.     

沙质荒漠化土地景观分类与制图—以毛乌素沙地为例

对景观分类理论和方法进行了探索,并以毛乌素沙地为研究对象,提出了一套适用于农牧交错区沙质荒漠化土地景观分类与制图的方法。由于沙丘分布广泛,与地带性植被和土壤的分布相迭加,增加了地表景观的破碎程度,使农牧交错区沙地的主要景观要素类型在结构上具有复合性。为了更好地描述沙地景观的复合性特征,作者提出景观基本结构组分的概念。首先确定景观基本结构组分,根据景观基本结构组分的不同组合确定景观要素类型;然后在野外调查的基础上,通过对遥感数据的解译和判读进行景观要素类型的划分;最后在GIS支持下成图。该方法对我国北方农牧交错区沙质荒漠化土地具有普遍意义。     

Detection and characterization of single biomolecules at surfaces

The investigation of bio-molecules has entered a new age since the development of methodologies capable of studies at the level of single molecules. In biology, most molecules show a complex dynamical behavior, with individual motions and transitions between different states, occurring as highly correlated in space and time within an arrangement of various elements. In order to resolve such dynamical changes in ensemble average techniques, one would have to synchronize all molecules, which is hard to achieve and might interfere with important system properties. Single molecule studies, in contrast, do not require pretreatment of the system and resume, therefore, much less invasive methodologies. Here, we review recent employments for the investigation of bio-molecules on surfaces, in which the high local and temporal resolution of two complementary techniques, atomic force microscopy and single molecule fluorescence microscopy, is used to address single molecules. Novel methodologies for the characterization of biologically relevant parameters, functions and dynamical aspects of individual molecules are described.     

Six Years of Criticality Assessments: What Have We Learned So Far?

The “criticality” of the various elements used in modern technologies is a topic of increasing interest, with groups from governments, consultancies, and academic institutions developing a variety of methodologies and using them to make assessments. Other groups from similar organizations are studying the methodologies that generate these assessments. Here, we analyze the different types of studies, review issues of methodology, and comment on features of nine different studies published between 2008 and mid‐2014. From these studies, we derive lists of problematic, debatable, and desirable aspects of criticality studies. We emphasize that the criticality of an element can vary depending on the target organization and that, because criticality is a dynamic state, it must be periodically re‐evaluated. There is substantial value to be derived if a more uniform methodology could be developed. We discuss how a harmonized methodological framework might be achieved and what its benefits could be. Putting into place such a structure for collaborative and publicly available criticality determinations would be very likely to better serve the present and future needs of corporations and governments than is the case at present, where different methodologies generate different results.     

Waves of genomic hitchhikers shed light on the evolution of gamebirds (Aves: Galliformes)

Background  

The phylogenetic tree of Galliformes (gamebirds, including megapodes, currassows, guinea fowl, New and Old World quails, chicken, pheasants, grouse, and turkeys) has been considerably remodeled over the last decades as new data and analytical methods became available. Analyzing presence/absence patterns of retroposed elements avoids the problems of homoplastic characters inherent in other methodologies. In gamebirds, chicken repeats 1 (CR1) are the most prevalent retroposed elements, but little is known about the activity of their various subtypes over time. Ascertaining the fixation patterns of CR1 elements would help unravel the phylogeny of gamebirds and other poorly resolved avian clades.     

Xenopus as a model system for vertebrate heart development

The African clawed frog, Xenopus laevis, is a valuable model system for studies of vertebrate heart development. In the following review, we describe a range of embryological and molecular methodologies that are used in Xenopus research and discuss key discoveries relating to heart development that have been made using this model system. We also discuss how the sequence of the Xenopus tropicalis genome provides a valuable tool for identification of orthologous genes and for identification of evolutionarily conserved promoter elements. Finally, both forward and reverse genetic approaches are currently being applied to Xenopus for the study of vertebrate heart development.     

Methods and applications for assembling large DNA constructs

The construction of large DNA molecules that encode pathways, biological machinery, and entire genomes has been limited to the reproduction of natural sequences. However, now that robust methods for assembling hundreds of DNA fragments into constructs > 20 kb are readily available, optimization of large genetic elements for metabolic engineering purposes is becoming more routine. Here, various DNA assembly methodologies are reviewed and some of their potential applications are discussed. We tested the potential of DNA assembly to install rational changes in complex biosynthetic pathways, their potential for generating complex libraries, and consider how various strategies are applicable to metabolic engineering.     

Purification and RNA binding properties of the polycytidylate-binding proteins alphaCP1 and alphaCP2

Regulation of mRNA turnover is a critical control mechanism of gene expression and is influenced by ribonucleoprotein (RNP) complexes that form on cis elements. All mRNAs have an intrinsic half-life and in many cases these half-lives can be altered by a variety of stimuli that are manifested through the formation or disruption of an RNP structure. The stability of alpha-globin mRNA is determined by elements in the 3' untranslated region that are bound by an RNP complex (alpha-complex) which appears to control the erythroid-specific accumulation of alpha-globin mRNA. The alpha-complex could consist of up to six distinct proteins or protein families. One of these families is a prominent polycytidylate binding activity which consists of two highly homologous proteins, alpha-complex proteins 1 and 2 (alphaCP1 and alphaCP2). This article focuses on various methodologies for the detection and manipulation of alphaCP1 and alphaCP2 binding to RNA and details means of isolating and characterizing mRNA bound by these proteins to study mRNA turnover and its regulation.     

The utility of transposable elements as tools for the isolation of plant genes

Transposable elements are segments of DNA which have the unique capability of being able to excise from one site in the genome and reintegrate into new, different sites elsewhere in the genome. When transposition takes place and integration occurs within a gene locus, mutations are frequently generated producing variegated or recessive phenotypes. This ability of transposable elements to act as mutagenic agents through their association with particular gene sequences has lead to the development of the procedure of transposon tagging or gene tagging in higher plants. Through this technique, transposable elements can be used to clone and isolate genes of interest for which little or nothing is known about the final product (i.e., polypeptide). This offers tremendous potential for the isolation of a variety of agronomically important genes, which are virtually impossible to recover by other currently available gene cloning methodologies. To date, the technique has been used successfully to isolate genes from corn and snapdragon. Using gene transfer technologies, the potential now exists to extend this approach to clone genes from other plant species. Advantages and limitations of transposon tagging for isolating plant genes will be discussed.     

依赖于5′端非编码区高级结构的真核生物mRNA翻译调控

翻译水平的调控是真核基因表达调控的重要环节.近年来的研究表明,许多真核基因的翻译依赖于RNA 5′端非编码区的结构元件.一些小结构元件,如铁离子反应元件,具有1个茎环结构,由铁离子介导控制转铁蛋白的翻译. 核糖开关通过结合特定代谢分子在2种结构状态下切换,调控可变剪接和翻译起始.另1个高度结构化的mRNA元件是内部核糖体进入位点,通过富集核糖体和起始因子促进基因的表达.本文综述了依赖于小结构元件、内部核糖体进入位点和核糖开关的真核基因翻译起始调控相应的研究成果和研究方法.对于研究的前景以及可能存在的挑战也作出阐述.     

Designing metal-peptide models for protein structure and function

Recent progress in the rational design of metal sites within peptide model systems shows increasing control in the placement of metals within helical bundles and inclusion of sophisticated elements such as second-sphere ligand interactions. A crystallographically characterized two-metal peptide model for diiron proteins represents a major achievement in de novo design methodologies. Increasingly complex and robust models for electron transfer through and between helices, and electrode-supported electron-transfer peptides, have been constructed. Design elements for peptide-supported ferredoxins and mononuclear Fe(II) and Zn(II) sites have been refined.     

Comparison of two methods for quality assessment of macroalgae assemblages,under different pollution types

The selection of adequate methodologies for the assessment of different biological quality elements is urgently needed for the application of the water framework directive (WFD 2000/60/EEC). In the case of macroalgae in coastal waters of the North East Atlantic, two methodologies have been proposed: the reduced species list (RSL) index and the quality of rocky bottoms (CFR) index. Both methods use multimetric approaches to evaluate the quality of macroalgae assemblages, which are based on community characteristics (species/populations richness, cover, percentage of opportunistic species, ecological state groups ratio, etc.). In this paper the results of applying both indices on three different types of pollution gradients in the North coast of Spain (bay of Biscay) are presented, in order to test their usefulness and intercalibration possibilities. In general terms, the CFR index responded more accurately than the RSL index to the pollution gradients under study. With respect to the indicators used in the current evaluation, richness, opportunistic species and cover seemed to be the most accurate for quality assessment of macroalgal communities. While the first two indicators are taken into account in both indices, the latter (cover) is only considered in the CFR index, even though the abundance of macroalgae is one of the aspects to be included in the evaluation of this biological element, according to the WFD.     

Recent advances in protein splicing: manipulating proteins in vitro and in vivo

Protein splicing is an intricate self-catalyzed protein rearrangement that converts an inactive protein precursor to biologically active proteins. In the past decade, mechanistic studies and extensive engineering of the naturally occurring protein splicing elements, termed inteins, has led to the development of numerous novel technologies. These intein-based methodologies permit in vitro and in vivo protein processing in ways previously not possible using traditional biochemical and genetic approaches. Inteins have been utilized in the production of protein and peptide arrays, as molecular switches and in the reconstitution of functional proteins by split-gene techniques.     

X-ray microanalysis of biological specimens by high voltage electron microscopy

For the purpose of analyzing and imaging chemical components of cells and tissues at the electron microscopic level, 3 fundamental methods are available, chemical, physical and biological. Among the physical methods, two methods qualifying and quantifying the elements in the structural components are very often employed. The first method is radioautography which can demonstrate the localization of radiolabeled compounds which were incorporated into cells and tissues after the administration of radiolabeled compounds. The second method is X-ray microanalysis which can qualitatively analyze and quantify the total amounts of elements present in cells and tissues. We have developed the two methodologies in combination with intermediate high or high voltage transmission electron microscopy (200–400 kV) and applied them to various kinds of organic and inorganic compounds present in biological materials. As for the first method, radioautography, I had already contributed a chapter to PHC (37/2). To the contrary, this review deals with another method, X-ray microanalysis, using semi-thin sections and intermediate high voltage electron microscopy developed in our laboratory.

X-ray microanalysis is a useful method to qualify and quantify basic elements in biological specimens. We first quantified the end-products of histochemical reactions such as Ag in radioautographs, Ce in phosphatase reaction and Au in colloidal gold immunostaining using semithin sections and quantified the reaction products observing by intermediate high voltage transmission electron microscopy at accelerating voltages from 100 to 400 kV. The P/B ratios of all the end products Ag, Ce and Au increased with the increase of the accelerating voltages from 100 to 400 kV. Then we analyzed various trace elements such as Zn, Ca, S and Cl which originally existed in cytoplasmic matrix or cell organelles of various cells, or such elements as Al which was absorbed into cells and tissues after oral administration, using both conventional chemical fixation and cryo-fixation followed by cryo-sectioning and freeze-drying, or freeze-substitution and dry-sectioning, or freeze-drying and dry-sectioning producing semithin sections similarly to radioautography. As the results, some trace elements which originally existed in cytoplasmic matrix or cell organelles of various cells in different organs such as Zn, Ca, S and Cl, were effectively detected. Zn was demonstrated in Paneth cell granules of mouse intestines and its P/B ratios showed a peak at 300 kV. Ca was found in human ligaments and rat mast cells with a maximum of P/B ratios at 350 kV. S and Cl were detected in mouse colonic goblet cells with maxima of P/B ratios at 300 kV. On the other hand, some elements which were absorbed by experimental administration into various cells and tissues in various organs, such as Al in lysosomes of hepatocytes and uriniferous tubule cells in mice was detected with a maximum of P/B ratios at 300 kV.

From the results, it was shown that X-ray microanalysis using semi-thin sections observed by intermediate high voltage transmission electron microscopy at 300–400 kV was very useful resulting in high P/B ratios for quantifying some trace elements in biological specimens. These methodologies should be utilized in microanalysis of various compounds and elements in various cells and tissues in various organs.     

High‐resolution elemental localization in vacuolate plant cells by nanoscale secondary ion mass spectrometry

By combining the capabilities of advanced sample preparation methodologies with the latest generation of secondary ion mass spectrometry instrumentation, we show that chemical information on the distribution of even dilute species in biological samples can be obtained with spatial resolutions of better than 100 nm. Here, we show the distribution of nickel and other elements in leaf tissue of the nickel hyperaccumulator plant Alyssum lesbiacum prepared by high‐pressure freezing and freeze substitution.