bluuebunny/crossref_metadata_2025 huggingface dataset:
THE ‘HEALTHCARE STATE’ IN TRANSITION
Authors: Kuhlmann Ellen , Burau Viola
Published: September 2008

The building of a European nation challenges entrenched ties between the state and the professions. Yet, in relation to healthcare, European law is especially weak and professional power is particularly strong. Against this background, the present paper aims to map out the specific configurations of the changing governance of healthcare and the dynamics arising from intersecting contexts of change. We argue that the ‘healthcare state’ is strongly shaped by national regulatory frameworks, while the platform for the power of a self-regulatory medical profession is increasingly international in nature. Across countries managerialism and performance measures together with evidence-based medicine and clinical guidelines are meant to improve the accountability of professionals and the safety of the public. However, the new regulatory tools may also have the opposite effect and indeed serve as a means to reassert professional power; here, the heightened internationalization of healthcare even provides new opportunities. We use material from different countries gathered in a number of research projects to assess the changing governance of healthcare and its contexts. The paper concludes by highlighting the significance of both national regulatory frameworks and (international) professionalism.
PUBLIC GOOD OR FREE MARKET?
Authors: Schnell Christiane
Published: September 2008

European continental countries traditionally emphasized the character of the cultural possessions as a public good. Now the technological development, accelerated expansion, internationalization, and the commercialization of the cultural economy bear a new quality of contradiction between social and economic demands in the field of cultural work. This problem is discussed with regard to the framework for freelance work in cultural professions in Germany. The main focus is on the specific institutional arrangement, which supports professionalism in the field of culture, without providing a privileged status comparable to the established professions. One central column of this arrangement is the German copy right law, which defines – in contrast to the Anglo-Saxon model, created as an investment protection – an individual property right of ‘intellectual property’ (‘Geistiges Eigentum’), against misuse, and economic exploitation. Against the background of the structural changes in the media in the 1990s, the EU is supporting the reform of copyright regulations in the European member states. It is claimed that present EU directions, developed within the concept of a ‘European knowledge-society’, break with the leading concept of cultural professions, but without providing a consistent foundation to solve the societal challenges of present culture and media production.
THE ‘REFLECTIVE CAPACITY’ OF PROFESSIONS CONFRONTED BY INTERNATIONAL COMPETITION
Authors: Champy Florent
Published: September 2008

This paper is devoted to the regulation of architectural activity in Europe and especially in France and to the ways architects manage this regulation. The globalization of this activity has generated a strong consciousness that supranational regulation is necessary, both among architects’ professional associations and in governments. One of the main difficulties is the differences in the division of labour from one country to another. But European regulation supposes that national states will implement the directives into national law. France is an example of the lightest possible adaptation. This is all the more surprising as the architectural profession in France is characterized by structural weaknesses to resist international competition and as the administration is willing to help it adapt to this competition. The history of trade unionism and the structure of representation explain some of the difficulties because of a particular trait of the architectural profession: what I call a weak ‘reflective capacity’. The paper is partly devoted to defining this notion and showing its relevance for the sociological analysis of professions. This weakness encourages individual strategies of adaptation to increasing international competition, without the professional associations and their representatives being able to control these strategies and their consequences for the profession. This demonstrates that the various sources of regulation are not exclusive and must be studied together so as to understand their interrelations.
PROFESSIONALS OF POLICIES FOR FIGHTING UNEMPLOYMENT IN FRANCE
Authors: Divay Sophie
Published: September 2008

The increase in unemployment that occurred in France at the end of the 1970s is one aspect that has favoured the creation of jobs specializing in the field of social welfare for the unemployed. This paper focuses on the analysis of the professional group of advisers specializing in job seeking. The first section traces the circumstances of the emergence of such a group and explains the heterogeneity of these advisers who seem, nevertheless, to have developed a specific common expertise and the same interpretation of their professionalism. In the second part, the activity of these professionals is situated in the larger framework of unemployed policies which result from a combination of international, European and national influences. The study of recent changes to unemployment policies reveals the effects of the ‘macro’ on the ‘micro’ level. In the third section, it is explained that new measures of social welfare for the unemployed tend to challenge the moral and deontological principles of these advisers whose professional identity and professionalism are challenged. These challenges to their identities are due to the intensification of the follow-up of the unemployed and to the application of bureaucratic, monitoring and measurement techniques and logic to assess the efficiency of the tools for promoting a quick return to work.
AP-2 factors act in concert with Notch to orchestrate terminal differentiation in skin epidermis
Authors: Wang Xuan , Pasolli H. Amalia , Williams Trevor , Fuchs Elaine
Published: October 2008

The mechanisms by which mammalian epidermal stem cells cease to proliferate and embark upon terminal differentiation are still poorly understood. By conditionally ablating two highly expressed transcription factors, AP-2α and AP-2γ, we unmasked functional redundancies and discovered an essential role for AP-2s in the process. In vivo and in vitro, AP-2 deficiency is accompanied by surprisingly minimal changes in basal gene expression but severely perturbed terminal differentiation and suppression of additional transcription factors and structural genes involved. In dissecting the underlying molecular pathways, we uncover parallel pathways involving AP-2 and Notch signaling, which converge to govern CCAAT/enhancer binding protein genes and orchestrate the transition from basal proliferation to suprabasal differentiation. Finally, we extend the striking similarities in compromising either Notch signaling or AP-2α/AP-2γ in developing skin to that in postnatal skin, where all hair follicles and sebaceous gland differentiation are also repressed and overt signs of premalignant conversion emerge.
Phylogenetic distances are encoded in networks of interacting pathways
Authors: Mazurie Aurélien , Bonchev Danail , Schwikowski Benno , Buck Gregory A.
Published: September 2008

Motivation: Although metabolic reactions are unquestionably shaped by evolutionary processes, the degree to which the overall structure and complexity of their interconnections are linked to the phylogeny of species has not been evaluated in depth. Here, we apply an original metabolome representation, termed Network of Interacting Pathways or NIP, with a combination of graph theoretical and machine learning strategies, to address this question. NIPs compress the information of the metabolic network exhibited by a species into much smaller networks of overlapping metabolic pathways, where nodes are pathways and links are the metabolites they exchange.

Results: Our analysis shows that a small set of descriptors of the structure and complexity of the NIPs combined into regression models reproduce very accurately reference phylogenetic distances derived from 16S rRNA sequences (10-fold cross-validation correlation coefficient higher than 0.9). Our method also showed better scores than previous work on metabolism-based phylogenetic reconstructions, as assessed by branch distances score, topological similarity and second cousins score. Thus, our metabolome representation as network of overlapping metabolic pathways captures sufficient information about the underlying evolutionary events leading to the formation of metabolic networks and species phylogeny. It is important to note that precise knowledge of all of the reactions in these pathways is not required for these reconstructions. These observations underscore the potential for the use of abstract, modular representations of metabolic reactions as tools in studying the evolution of species.

Contact:  aurelien.mazurie@pasteur.fr

Supplementary information:  Supplementary data are available at Bioinformatics online.
MSX-3D: a tool to validate 3D protein models using mass spectrometry
Authors: Heymann Michaël , Paramelle David , Subra Gilles , Forest Eric , Martinez Jean , Geourjon Christophe , Deléage Gilbert
Published: October 2008

Motivation: The technique of chemical cross-linking followed by mass spectrometry has proven to bring valuable information about the protein structure and interactions between proteic subunits. It is an effective and efficient way to experimentally investigate some aspects of a protein structure when NMR and X-ray crystallography data are lacking.

Results: We introduce MSX-3D, a tool specifically geared to validate protein models using mass spectrometry. In addition to classical peptides identifications, it allows an interactive 3D visualization of the distance constraints derived from a cross-linking experiment.

Availability: Freely available at http://proteomics-pbil.ibcp.fr

Contact:  g.deleage@ibcp.fr
A Chinese Herbal Preparation Containing Radix Salviae Miltiorrhizae, Radix Notoginseng and Borneolum Syntheticum Reduces Circulating Adhesion Molecules
Authors: O’Brien Kylie A. , Ling Shanhong , Abbas Estelle , Dai Aozhi , Zhang Jiansheng , Wang Wen Cheng , Bensoussan Alan , Luo Ruizhi , Guo Zhi-Xin , Komesaroff Paul A.
Published: September 2008

Circulating adhesion molecules (CAMs), surface proteins expressed in the vascular endothelium, have emerged as risk factors for cardiovascular disease (CVD). CAMs are involved in intercellular communication that are believed to play a role in atherosclerosis. A Chinese medicine, the “Dantonic Pill” (DP) (also known as the “Cardiotonic Pill”), containing three Chinese herbal material medica, Radix Salviae Miltiorrhizae, Radix Notoginseng and Borneolum Syntheticum, has been used in China for the prevention and management of CVD. Previous laboratory and animal studies have suggested that this preparation reduces both atherogenesis and adhesion molecule expression. A parallel double blind randomized placebo‐controlled study was conducted to assess the effects of the DP on three species of CAM (intercellular cell adhesion molecule‐1 (ICAM‐1), vascular cell adhesion molecule‐1 and endothelial cell selectin (E‐selectin)) in participants with mild‐moderate hypercholesterolemia. Secondary endpoints included biochemical and hematological variables and clinical effects. Forty participants were randomized to either treatment or control for 12 weeks. Treatment with DP was associated with a statistically significant decrease in ICAM‐1 (9% decrease, P = .03) and E‐Selectin (15% decrease, P = .004). There was no significant change in renal function tests, liver function tests, glucose, lipids or C‐reactive protein levels and clinical adverse effects did not differ between the active and the control groups. There were no relevant changes in participants receiving placebo. These results suggest that this herbal medicine may contribute to the development of a novel approach to cardiovascular risk reduction.
The Influence of Acupressure at Extra 1 Acupuncture Point on the Spectral Entropy of the EEG and the LF/HF Ratio of Heart Rate Variability
Authors: Arai Young-Chang P. , Ushida Takahiro , Matsubara Takako , Shimo Kazuhiro , Ito Hiroshi , Sato Yuko , Wakao Yoshiko , Komatsu Toru
Published: October 2008

Acupressure applied on the Extra 1 acupuncture point results in sedation, thereby reducing bispectral index (BIS) values. Mental status and hypnotic agents influence the autonomic nervous system. We hypothesized that acupressure at the Extra 1 point would induce sedation and change sympatho‐parasympathetic nerve balance. We investigated the effect of acupressure at the Extra 1 point on the EEG spectral entropy values and heart rate variability (HRV). Forty‐eight volunteers (24 males and 24 females) were randomly assigned to the control or Extra 1 group. The control group received acupressure at a sham point and the Extra 1 group received acupressure at the Extra 1 point. Acupressure was applied for 5 min. The record of the EEG spectral entropy values and HRV started 5 min before acupressure and stopped 5 min after acupressure. Acupressure significantly reduced the EEG spectral entropy values in both groups, but the values of the Extra 1 group were significantly smaller than those of the control group (P < .01). Acupressure significantly decreased the LF/HF ratio of HRV in both groups (P < .05). When divided upon gender, although acupressure tended to decrease the LF/HF ratio, the ratio significantly decreased during and after acupressure only in females of the Extra 1 group (P < .05). We concluded that acupressure on the Extra 1 point significantly reduced the EEG spectral entropy in both the genders, but affected the LF/HF ratio of HRV only in females.
Hierarchical Bayesian approach for population dynamics modelling of fish complexes without species-specific data
Authors: Jiao Yan , Hayes Christopher , Cortés Enric
Published: September 2008

Jiao, Y., Hayes, C., and Cortés, E. 2009. Hierarchical Bayesian approach for population dynamics modelling of fish complexes without species-specific data. – ICES Journal of Marine Science, 66: 367–377. Modelling the population dynamics of fish complexes is challenging, and many species have been assessed and managed as a complex that was treated as a single species. Two Bayesian state-space surplus production models with multilevel priors (hierarchical models) were developed to simulate variability in population growth rates of species in a complex, using the hammerhead shark complex (Sphyrna spp.) of the Atlantic and Gulf of Mexico coasts of the US as an example. The complex consists of three species: scalloped (Sphyrna lewini), great (Sphyrna mokarran), and smooth hammerhead (Sphyrna zygaena). Bayesian state-space surplus production models with multilevel priors fitted the hammerhead data better than a model based on single-level priors. The hierarchical Bayesian approach represents an intermediate strategy between traditional models that do not include variability among species, and highly parameterized models that assign an estimate of parameters to each species. By ignoring the variability among species, confidence intervals of the estimates of stock status indicators can be unrealistically narrow, possibly leading to high-risk management strategies being adopted. Use of multilevel priors in a hierarchical Bayesian approach is suggested for future hammerhead shark stock assessments and for modelling fish complexes lacking species-specific data.
Structure and properties of strontium-doped phosphate-based glasses
Authors: Abou Neel Ensanya A , Chrzanowski Wojciech , Pickup David M , O'Dell Luke A , Mordan Nicola J , Newport Robert J , Smith Mark E , Knowles Jonathan C
Published: October 2008

Owing to similarity in both ionic size and polarity, strontium (Sr
2+
) is known to behave in a comparable way to calcium (Ca
2+
), and its role in bone metabolism has been well documented as both anti-resorptive and bone forming. In this study, novel quaternary strontium-doped phosphate-based glasses, containing 1, 3 and 5 mol% SrO, were synthesized and characterized.
31
P magic angle spinning (MAS) nuclear magnetic resonance results showed that, as the Sr
2+
content is increased in the glasses, there is a slight increase in disproportionation of
Q
2
phosphorus environments into
Q
1
and
Q
3
environments. Moreover, shortening and strengthening of the phosphorus to bridging oxygen distance occurred as obtained from FTIR. The general broadening of the spectral features with Sr
2+
content is most probably due to the increased variation of the phosphate–cation bonding interactions caused by the introduction of the third cation. This increased disorder may be the cause of the increased degradation of the Sr-containing glasses relative to the Sr-free glass. As confirmed from elemental analysis, all Sr-containing glasses showed higher Na
2
O than expected and this also could be accounted for by the higher degradation of these glasses compared with Sr-free glasses. Measurements of surface free energy (SFE) showed that incorporation of strontium had no effect on SFE, and samples had relatively higher fractional polarity, which is not expected to promote high cell activity. From viability studies, however, the incorporation of Sr
2+
showed better cellular response than Sr
2+
-free glasses, but still lower than the positive control. This unfavourable cellular response could be due to the high degradation nature of these glasses and not due to the presence of Sr
2+
.
The diverse origins of New Zealand house mice
Authors: Searle Jeremy B , Jamieson Paul M , Gündüz İslam , Stevens Mark I , Jones Eleanor P , Gemmill Chrissen E.C , King Carolyn M
Published: October 2008

Molecular markers and morphological characters can help infer the colonization history of organisms. A combination of mitochondrial (mt)d-loop DNA sequences, nuclear DNA data, external measurements and skull characteristics shows that house mice (Mus musculus) in New Zealand and its outlying islands are descended from very diverse sources. The predominant genome isMus musculus domesticus(from western Europe), butMus musculus musculus(from central Europe) andMus musculus castaneus(from southern Asia) are also represented genetically. These subspecies have hybridized to produce combinations ofmusculusanddomesticusnuclear DNA coupled withdomesticusmtDNA, andcastaneusormusculusmtDNA withdomesticusnuclear DNA. The majority of the mice withdomesticusmtDNA that we sampled hadd-loop sequences identical to two haplotypes common in Britain. This is consistent with long-term British–New Zealand cultural linkages. The origins of thecastaneusmtDNA sequences widespread in New Zealand are less easy to identify.
Inferring malaria parasite population structure from serological networks
Authors: Buckee Caroline O , Bull Peter C , Gupta Sunetra
Published: October 2008

The malaria parasite
Plasmodium falciparum
is characterized by high levels of genetic diversity at antigenic loci involved in virulence and immune evasion. Knowledge of the population structure and dynamics of these genes is important for designing control programmes and understanding the acquisition of immunity to malaria; however, high rates of homologous and non-homologous recombination as well as complex patterns of expression within hosts have hindered attempts to elucidate these structures experimentally. Here, we analyse serological data from Kenya using a novel network technique to deconstruct the relationships between patients' immune responses to different parasite isolates. We show that particular population structures and expression patterns produce distinctive signatures within serological networks of parasite recognition, which can be used to discriminate between competing hypotheses regarding the organization of these genes. Our analysis suggests that different levels of immune selection occur within different groups of the same multigene family leading to mixed population structures.
Assessing visual requirements for social context-dependent activation of the songbird song system
Authors: Hara Erina , Kubikova Lubica , Hessler Neal A , Jarvis Erich D
Published: October 2008

Social context has been shown to have a profound influence on brain activation in a wide range of vertebrate species. Best studied in songbirds, when males sing undirected song, the level of neural activity and expression of immediate early genes (IEGs) in several song nuclei is dramatically higher or lower than when they sing directed song to other birds, particularly females. This differential social context-dependent activation is independent of auditory input and is not simply dependent on the motor act of singing. These findings suggested that the critical sensory modality driving social context-dependent differences in the brain could be visual cues. Here, we tested this hypothesis by examining IEG activation in song nuclei in hemispheres to which visual input was normal or blocked. We found that covering one eye blocked visually induced IEG expression throughout both contralateral visual pathways of the brain, and reduced activation of the contralateral ventral tegmental area, a non-visual midbrain motivation-related area affected by social context. However, blocking visual input had no effect on the social context-dependent activation of the contralateral song nuclei during female-directed singing. Our findings suggest that individual sensory modalities are not direct driving forces for the social context differences in song nuclei during singing. Rather, these social context differences in brain activation appear to depend more on the general sense that another individual is present.
Accretion and core formation: constraints from metal–silicate partitioning
Authors: Wood Bernard J
Published: October 2008

Experimental metal–silicate partitioning data for Ni, Co, V, Cr, Nb, Mn, Si and W were used to investigate the geochemical consequences of a range of models for accretion and core formation on Earth. The starting assumptions were chondritic ratios of refractory elements in the Earth and the segregation of metal at the bottom of a magma ocean, which deepened as the planet grew and which had, at its base, a temperature close to the liquidus of the silicate. The models examined were as follows. (i) Continuous segregation from a mantle which is chemically homogeneous and which has a fixed oxidation state, corresponding to 6.26 per cent oxidized Fe. Although Ni, Co and W partitioning is consistent with chondritic ratios, the current V content of the silicate Earth cannot be reconciled with core segregation under these conditions of fixed oxidation state. (ii) Continuous segregation from a mantle which is chemically homogeneous but in which the Earth became more oxidized as it grew. In this case, the Ni, Co, W, V, Cr and Nb contents of core and mantle are easily matched to those calculated from the chondritic ratios of refractory elements. The magma ocean is calculated to maintain a thickness approximately 35 per cent of the depth to the core–mantle boundary in the accreting Earth, yielding a maximum pressure of 44 GPa. This model yields a Si content of the core of 5.7 per cent, in good agreement with cosmochemical estimates and with recent isotopic data. (iii) Continuous segregation from a mantle which is not homogeneous and in which the core equilibrates with a restricted volume of mantle at the base of the magma ocean. This is found to increase depth of the magma ocean by approximately 50 per cent. All of the other elements (except Mn) have partitioning consistent with chondritic abundances in the Earth, provided the Earth became, as before, progressively oxidized during accretion. (iv) Continuous segregation of metal from a crystal-melt mush. In this case, pressures decrease to a maximum of 31 GPa and it is extremely difficult to match the calculated mantle contents of the highly incompatible elements Nb and W to those observed. Progressive oxidation is required to fit the observed mantle contents of vanadium. All of the scenarios discussed above point to progressive oxidation having occurred as the Earth grew. The Earth appears to be depleted in Mn relative to the chondritic reference.
Early differentiation of the Earth and the Moon
Authors: Bourdon Bernard , Touboul Mathieu , Caro Guillaume , Kleine Thorsten
Published: October 2008

We examine the implications of new182W and142Nd data for Mars and the Moon for the early evolution of the Earth. The similarity of182W in the terrestrial and lunar mantles and their apparently differing Hf/W ratios indicate that the Moon-forming giant impact most probably took place more than 60 Ma after the formation of calcium-aluminium-rich inclusions (4.568 Gyr). This is not inconsistent with the apparent U–Pb age of the Earth. The new142Nd data for Martian meteorites show that Mars probably has a super-chondritic Sm/Nd that could coincide with that of the Earth and the Moon. If this is interpreted by an early mantle differentiation event, this requires a buried enriched reservoir for the three objects. This is highly unlikely. For the Earth, we show, based on new mass-balance calculations for Nd isotopes, that the presence of a hidden reservoir is difficult to reconcile with the combined142Nd–143Nd systematics of the Earth's mantle. We argue that a likely possibility is that the missing component was lost during or prior to accretion. Furthermore, the142Nd data for the Moon that were used to argue for the solidification of the magma ocean atca200 Myr are reinterpreted. Cumulate overturn, magma mixing and melting following lunar magma ocean crystallization at 50–100 Myr could have yielded the 200 Myr model age.
The redox state of the mantle during and just after core formation
Authors: Frost D.J , Mann U , Asahara Y , Rubie D.C
Published: October 2008

Siderophile elements are depleted in the Earth's mantle, relative to chondritic meteorites, as a result of equilibration with core-forming Fe-rich metal. Measurements of metal–silicate partition coefficients show that mantle depletions of slightly siderophile elements (e.g. Cr, V) must have occurred at more reducing conditions than those inferred from the current mantle FeO content. This implies that the oxidation state (i.e. FeO content) of the mantle increased with time as accretion proceeded. The oxygen fugacity of the present-day upper mantle is several orders of magnitude higher than the level imposed by equilibrium with core-forming Fe metal. This results from an increase in the Fe
2
O
3
content of the mantle that probably occurred in the first 1 Ga of the Earth's history. Here we explore fractionation mechanisms that could have caused mantle FeO and Fe
2
O
3
contents to increase while the oxidation state of accreting material remained constant (homogeneous accretion). Using measured metal–silicate partition coefficients for O and Si, we have modelled core–mantle equilibration in a magma ocean that became progressively deeper as accretion proceeded. The model indicates that the mantle would have become gradually oxidized as a result of Si entering the core. However, the increase in mantle FeO content and oxygen fugacity is limited by the fact that O also partitions into the core at high temperatures, which lowers the FeO content of the mantle. (Mg,Fe)(Al,Si)O
3
perovskite, the dominant lower mantle mineral, has a strong affinity for Fe
2
O
3
even in the presence of metallic Fe. As the upper mantle would have been poor in Fe
2
O
3
during core formation, FeO would have disproportionated to produce Fe
2
O
3
(in perovskite) and Fe metal. Loss of some disproportionated Fe metal to the core would have enriched the remaining mantle in Fe
2
O
3
and, if the entire mantle was then homogenized, the oxygen fugacity of the upper mantle would have been raised to its present-day level.
Signal integration in bacterial two-component regulatory systems
Authors: Mitrophanov Alexander Y. , Groisman Eduardo A.
Published: October 2008

Two-component systems (TCSs) and phosphorelays are key mediators of bacterial signal transduction. The signals activating these systems promote the phosphorylated state of a response regulator, which is generally the form that carries out specific functions such as binding to DNA and catalysis of biochemical reactions. An emerging class of proteins—termed TCS connectors—modulate the output of TCSs by affecting the phosphorylation state of response regulators. TCS connectors use different mechanisms of action for signal integration, as well as in the coordination and fine-tuning of cellular processes. Present in both Gram-positive and Gram-negative bacteria, TCS connectors are critical for a variety of physiological functions including sporulation, competence, antibiotic resistance, and the transition to stationary phase.
Proteoglycan desulfation determines the efficiency of chondrocyte autophagy and the extent of FGF signaling during endochondral ossification
Authors: Settembre Carmine , Arteaga-Solis Emilio , McKee Marc D. , de Pablo Raquel , Al Awqati Qais , Ballabio Andrea , Karsenty Gerard
Published: October 2008

Cartilage extracellular matrix (ECM) contains large amounts of proteoglycans made of a protein core decorated by highly sulfated sugar chains, the glycosaminoglycans (GAGs). GAGs desulfation, a necessary step for their degradation, is exerted by sulfatases that are activated by another enzyme, Sulfatase-Modifying Factor 1 (SUMF1), whose inactivation in humans leads to severe skeletal abnormalities. We show here that despite being expressed in both osteoblasts and chondrocytes Sumf1 does not affect osteoblast differentiation. Conversely, in chondrocytes it favors ECM production and autophagy and promotes proliferation and differentiation by limiting FGF signaling. Thus, proteoglycan desulfation is a critical regulator of chondrogenesis.
Meeting DNA palindromes head-to-head
Authors: Smith Gerald R.
Published: October 2008

Particular DNA sequences have long been known to have exceptional structures and biological properties. Famous in the medical world are the trinucleotide repeat sequences, such as (CTG)n, and their association with more than a dozen neurodegenerative diseases. Numerous meetings have been held to discuss these repeats and the diseases they cause. Now, a much-needed meeting has been held to discuss other noncanonical (non-B-form) DNA structures, their properties, and their biological consequences. Although the meeting was titled “DNA palindromes: roles, consequences, and implications of structurally ambivalent DNA,” the participants discussed and debated a range of additional structures—dubbed “Z,” “HJ,” “G4,” and “H” DNA—as well as trinucleotide repeats. These remarkable structures can have profound effects on chromosomes and organisms, ranging from mutational hotspots in bacteria to causes of intellectual disability in humans. Bringing together four dozen researchers prominent in the field focused attention on these controversial DNA structures in a way that promises to spur greater understanding of DNA elements critical to life and health.
Deconstructing meiosis one kinase at a time: polo pushes past pachytene
Authors: Hollingsworth Nancy M.
Published: October 2008

The transition from pachytene to Meiosis I is a key regulatory point in yeast meiosis. This transition requires Ndt80, a transcription factor that commits cells to complete meiosis by expression of a diverse set of genes. In this issue of Genes & Development, Sourirajan and Lichten (2627-2632) report that CDC5, an NDT80-regulated gene encoding yeast polo-like kinase, is sufficient for Holliday junction resolution and exit from pachytene. Determining the meiotic targets of Cdc5 therefore provides a new approach for identifying a eukaryotic Holliday resolvase.
Sequestration of Polo kinase to microtubules by phosphopriming-independent binding to Map205 is relieved by phosphorylation at a CDK site in mitosis
Authors: Archambault Vincent , D’Avino Pier Paolo , Deery Michael J. , Lilley Kathryn S. , Glover David M.
Published: October 2008

The conserved Polo kinase controls multiple events in mitosis and cytokinesis. Although Polo-like kinases are regulated by phosphorylation and proteolysis, control of subcellular localization plays a major role in coordinating their mitotic functions. This is achieved largely by the Polo-Box Domain, which binds prephosphorylated targets. However, it remains unclear whether and how Polo might interact with partner proteins when priming mitotic kinases are inactive. Here we show that Polo associates with microtubules in interphase and cytokinesis, through a strong interaction with the microtubule-associated protein Map205. Surprisingly, this interaction does not require priming phosphorylation of Map205, and the Polo-Box Domain of Polo is required but not sufficient for this interaction. Moreover, phosphorylation of Map205 at a CDK site relieves this interaction. Map205 can stabilize Polo and inhibit its cellular activity in vivo. In syncytial embryos, the centrosome defects observed in polo hypomorphs are enhanced by overexpression of Map205 and suppressed by its deletion. We propose that Map205-dependent targeting of Polo to microtubules provides a stable reservoir of Polo that can be rapidly mobilized by the activity of Cdk1 at mitotic entry.
The American Wandering Spider Cupiennius salei
Authors: Prpic Nikola-Michael , Schoppmeier Michael , Damen Wim G.M.
Published: October 2008

INTRODUCTION

The spider Cupiennius salei is a useful laboratory model for embryological and physiological studies. Its highly developed sensory organs also make it an excellent model for behavioral studies. Furthermore, Cupiennius has contributed greatly to the study of evolutionary developmental questions. This chelicerate arthropod is particularly useful for such studies because of its phylogenetic position and the availability of tools to study and manipulate its embryonic development.
Opossum (Monodelphis domestica): A Marsupial Development Model
Authors: Keyte Anna L. , Smith Kathleen K.
Published: October 2008

INTRODUCTION

Monodelphis domestica is the most commonly used laboratory marsupial. In addition to the many factors that make it a convenient laboratory animal (small size, ease of care, nonseasonal breeding), it is the first marsupial whose genome has been sequenced. In this article, we present an overview of aspects of its biology and its use as a model organism. We also discuss basic care, breeding, embryo manipulation, and modifications of common techniques for the study of the development of this species.
High-Throughput Cloning of Open Reading Frames (ORFs): Assembling Large Sets of Expression Constructs
Authors: Link Andrew J. , LaBaer Joshua
Published: October 2008

INTRODUCTION

Nearly all methods for studying protein function begin with the expression of protein from cloned copies of the protein-coding sequences. Thus, obtaining a complete set of validated protein-coding clones is the first step toward establishing a functional proteomics platform for any organism of interest. Different protein functional studies demand different protein expression vectors; therefore, a flexible vector system should be used that enables the cloned sequences to be transferred rapidly to any vector. This can be achieved most efficiently by using vectors that employ recombinational cloning, a strategy that allows DNA fragments flanked by site-specific recombination sites to be moved from one vector to another in a single-step procedure, in frame and without mutation. There is now extensive experience with several such systems, including the Gateway system (Invitrogen) described here. These reactions are simple enough to allow high-throughput (HT) automation and are highly efficient. Once a “master” clone is created, the identical sequences can be transferred easily into all bacterial, mammalian, and viral vectors commonly used for protein analysis in vivo or in vitro. This protocol describes the process of transferring protein-coding sequences from a master vector into a protein expression (destination) vector for use in functional experiments.
Propagating Antirrhinum
Authors: Hudson Andrew , Critchley Joanna , Erasmus Yvette
Published: October 2008

INTRODUCTION

This protocol describes general strategies for propagating Antirrhinum (snapdragon) species: self- and cross-pollination, cuttings, and grafting. Antirrhinum majus cultivars and some wild species are self-fertile, but they require self-pollination for high seed yields. Although self-fertile, A. majus shows unilateral incompatibility and can only be crossed to other self-incompatible species as the female parent. All Antirrhinum species can be propagated clonally from cuttings. Antirrhinum also readily forms grafts within and between species.
Establishing and Maintaining a Colony of Planarians
Authors: Oviedo Néstor J. , Nicolas Cindy L. , Adams Dany S. , Levin Michael
Published: October 2008

INTRODUCTION

To provide sufficient material for experimentation, a laboratory needs to expand and maintain a colony of planarians. It is crucial to keep a stable, healthy population of animals in a consistent environment to avoid inter-animal variability and modifier effects that can mask true phenotypes from experimental perturbation. In this protocol, we describe basic procedures for establishing and maintaining healthy colonies of Dugesia japonica, Schmidtea mediterranea, and Girardia tigrina (commonly found in the wild and commercially available in the United States). Although the recommendations are based on our optimization of conditions for G. tigrina, many of the procedures (such as food preparation and feeding strategy) can be applied to other species. For best results, the culture water must be carefully monitored and adjusted for each species.
Gene Knockdown in Planarians Using RNA Interference
Authors: Oviedo Néstor J. , Nicolas Cindy L. , Adams Dany S. , Levin Michael
Published: October 2008

INTRODUCTION

This protocol describes how to produce gene knockdown in planarians using RNA interference (RNAi). It is a standard technique to evaluate gene function during regeneration and tissue maintenance in planarians. The procedure involves microinjecting double-stranded RNA (dsRNA) synthesized in vitro. Depending on the gene target, this technique can produce robust phenotypes that can be further evaluated by diverse macroscopic or microscopic procedures.
Live Imaging of Planarian Membrane Potential Using DiBAC4(3): Figure 1.
Authors: Oviedo Néstor J. , Nicolas Cindy L. , Adams Dany S. , Levin Michael
Published: October 2008

INTRODUCTION

This protocol describes how to use the anionic membrane voltage-reporting dye DiBAC4(3) to generate images of cell membrane potential in live planarians. These images qualitatively reveal variations in time-averaged membrane potential across different regions of the organism. Changes in these images due to experimental treatments reveal how the particular treatment affects this physiological parameter. This method is a great improvement over standard electrophysiological techniques, which cannot be used to gain an understanding of the electrical properties of an entire worm or a regenerating fragment, due to small cell size and large cell number. When the proper controls are performed, this technique is a very powerful and simple way to gather physiologic data.
Assessment of the Olfactory Response to Chemicals or Bacteria in Pristionchus Nematodes
Authors: Rae Robbie , Schlager Benjamin , Sommer Ralf J.
Published: October 2008

INTRODUCTION

In the soil environment, nematodes must rely on a number of host-specific chemical cues in order to find potential beetle hosts. They must also discriminate among different food choices (i.e., bacteria), which is important because if the nematodes concentrate on an unsuitable food source, they may die. To detect the bacteria and host-specific chemicals, nematodes use sensory structures called amphids and phasmids that are located on the head and tail, respectively. The olfactory response of nematodes can be studied in the laboratory using the very simple agar-based assay described here. This assay determines the attractiveness of Pristionchus to a range of beetle-associated compounds and has allowed detailed molecular mechanisms of olfaction to be studied in Pristionchus. It can also be used to examine the food choice of these nematodes when they are faced with a number of bacteria.
qRT-PCR of Microbial Biofilms
Authors: Pérez-Osorio Ailyn C. , Franklin Michael J.
Published: October 2008

INTRODUCTION

Bacteria growing in biofilms often express a different subset of genes compared to the same strains growing planktonically. Quantitative reverse transcriptase real time PCR (qRT-PCR) can be used effectively to quantify the number of RNA transcripts of specific genes from bacteria growing in biofilms. qRT-PCR has a large dynamic range and may be used to verify gene expression data obtained from microarrays. In addition, qRT-PCR is sensitive, and therefore may be used to quantify gene expression from biofilm samples where only a small amount of biological material is available, as in samples obtained by laser capture microdissection microscopy (LCMM). The most commonly used qRT-PCR methods are the SYBR Green and dual-labeled probe (Taqman) approaches. Both approaches use reverse transcription to convert mRNA to cDNA, followed by PCR amplification of the cDNA. This article describes steps involved in aspects of qRT-PCR including (1) primer design, (2) primer and probe performance testing, (3) qRT-PCR using the Corbett Rotor-Gene system, and (4) data export and analysis.
Collection and Fixation of Spider Embryos
Authors: Prpic Nikola-Michael , Schoppmeier Michael , Damen Wim G.M.
Published: October 2008

INTRODUCTION

The spider Cupiennius salei, commonly known as the American wandering spider, is a particularly useful laboratory model for embryological studies because of the availability of tools to study and manipulate its embryonic development. Cupiennius is used to study axis formation, segmentation, appendage development, neurogenesis, and silk production. These studies contribute to our understanding of the evolution of these processes, but they also help us to understand the origin and diversification of evolutionary novelties. Comparisons between spiders and insects can show the degree of conservation and divergence of developmental mechanisms during arthropod evolution. Any embryological feature conserved between spiders and insects is likely to represent an ancestral feature for arthropods. Comparative molecular embryological work in insects and spiders should eventually allow us to define a molecular archetype for the phylum Arthropoda. This in itself will be a necessary cornerstone for comparing the different metazoan phyla, including chordates. This protocol describes the collection and fixation of embryos from C. salei. The fixed embryos can be stored at -20°C for prolonged periods and used for in situ hybridization, in studies of apoptosis using terminal deoxynucleotidyl-transferase-mediated dUTP-digoxigenin nick-end labeling (TUNEL), and for immunohistochemistry.
Gene Silencing via Embryonic RNAi in Spider Embryos
Authors: Prpic Nikola-Michael , Schoppmeier Michael , Damen Wim G.M.
Published: October 2008

INTRODUCTION

The spider Cupiennius salei, commonly known as the American wandering spider, is a particularly useful laboratory model for embryological studies because of the availability of tools to study and manipulate its embryonic development. Cupiennius is used to study axis formation, segmentation, appendage development, neurogenesis, and silk production. These studies contribute to our understanding of the evolution of these processes, but they also help us to understand the origin and diversification of evolutionary novelties. Comparisons between spiders and insects can show the degree of conservation and divergence of developmental mechanisms during arthropod evolution. Any embryological feature conserved between spiders and insects is likely to represent an ancestral feature for arthropods. Comparative molecular embryological work in insects and spiders should eventually allow us to define a molecular archetype for the phylum Arthropoda. This in itself will be a necessary cornerstone for comparing the different metazoan phyla, including chordates. The discovery of RNA interference (RNAi), in which double-stranded RNA (dsRNA) suppresses the translation of homologous mRNA, has had a huge impact on evolutionary developmental biology by enabling the analysis of loss-of-function phenotypes in organisms in which classical genetic analysis is laborious or not possible. This protocol describes the application of RNAi to embryos of the spider C. salei.
Monodelphis Whole-Embryo Culture
Authors: Keyte Anna L. , Smith Kathleen K.
Published: October 2008

INTRODUCTION

Monodelphis domestica, the gray, short-tailed, or laboratory opossum, is the most commonly used laboratory marsupial. In addition to the factors that make it a convenient laboratory animal (small size, ease of care, nonseasonal breeding), it is the first marsupial whose genome has been sequenced. Monodelphis has proven useful as a model organism for studies on spinal cord regeneration, ultraviolet (UV)-induced melanoma, and genetic influences on cholesterol, as well as comparative studies of the immune system. In addition, Monodelphis has been used to understand the basic functions of the olfactory system and the role of various olfactory chemicals in social and reproductive behavior. Recently, Monodelphis has been used to understand fundamental aspects of marsupial development, anatomy, evolution, and evolutionary consequences of the derived marsupial mode of development and reproduction. The embryos of Monodelphis, like those of other marsupials, can be cultured in vitro. The length of embryo viability depends in part on the stage at which culture begins, but embryos of different species of marsupials have been cultured for 18 h to almost 72 h. Good culture results for Monodelphis have been obtained using the method presented here. Embryos can be manipulated and then placed in the incubator. We have applied this technique most commonly to embryos at stages 23-25; they have retained viability and normal development through stage 26 when embryos would begin to implant in vivo.
The leader as motivator: coach and self‐esteem builder
Authors: Castelli Patricia Ann
Published: September 2008

Purpose

The purpose of this paper is to determine what motivating strategies followers (comprised of low and high need achievers) view as most necessary from their leaders.

Design/methodology/approach

Three survey instruments were administered to 297 working professionals taking business courses offered through a college of management at a private university. Correlational research methodology was used to investigate the relationships between self‐attributed needs for achievement on demographic, interest and effort variables. Discriminant analysis was used to determine if motivational and profile characteristics could be used to predict low and high self‐attributed need achievers.

Findings

The major findings of this study centered on the leader's ability to inspire followers by using a variety of interesting coaching techniques and by building their self‐esteem regardless of achievement orientation.

Research limitations/implications

The results of this study are tentative and need to be reproduced with a larger, more comprehensive sample of a broader population.

Practical implications

This research offers leaders practical techniques for effectively utilizing motivating strategies which may increase their followers' interest and effort in a performance situation.

Originality/value

The findings of this study indicating that interest and effort can be used to improve motivation suggest that followers may exhibit significant gains in continuing motivation when relevant techniques are applied by the leader.