bluuebunny/crossref_metadata_2025_split huggingface dataset:
CTAB-PAGE
Authors: Simpson Richard J. 
Published: April 2010

INTRODUCTIONAlthough SDS-PAGE is the method of choice for most denaturing gel electrophoresis procedures, the anionic detergent SDS still presents some drawbacks. For example, SDS forms crystals at low temperatures and, in some cases, causes proteins to aggregate or precipitate. In addition, some proteins are not well-resolved in SDS gels or may migrate anomalously. In these situations, the use of a cationic detergent for PAGE offers an alternative approach. The system described in this protocol uses the cationic detergent cetyltrimethyl ammonium bromide (CTAB) and includes a stacking gel based on the zwitterion arginine (used as a stacking agent) and tricine (N-tris[hydroxymethyl]-methylglycine) used as a counterion and buffer. Some proteins separated on the CTAB electrophoresis system retain their native enzymatic activity, provided the samples are prepared without boiling and without the addition of a reducing agent.
Rapid Coomassie Blue Staining of Protein Gels
Authors: Simpson Richard J. 
Published: April 2010

INTRODUCTIONCoomassie brillant blue R250 (CBR-250) and silver staining are the most widely used methods for the routine visualization of proteins separated by SDS-PAGE. CBR-250 is an organic dye that complexes with basic amino acids, such as arginine, lysine, and histidine, as well as tyrosine. Conventional CBR-250 staining is capable of detecting as little as 30-100 ng of protein, but sensitivity can be improved by performing the staining and destaining at elevated temperatures. The method described in this protocol, which is a modified version of the conventional Coomassie protocol, speeds up the destaining process for faster results with increased sensitivity and is compatible with mass-spectrometry-based methods for identifying proteins.
Multiply Primed Rolling-Circle Amplification Method for the Amplification of Circular DNA Viruses
Authors: Stevens Hans , Rector Annabel , Van Ranst Marc 
Published: April 2010

INTRODUCTIONThe use of whole genome amplification and analysis of viruses is of increasing importance, as data generated using these methods are currently used for clinical diagnostics, epidemiological studies, phylogenetic analyses, and studies of genome organization and evolution. The best known amplification method for DNA is the polymerase chain reaction (PCR). This technique, however, has drawbacks: PCR produces relatively small amplicons and also requires prior knowledge of sequence data for the construction of the required consensus or degenerate primers. For circular DNA templates, it is possible to overcome these drawbacks by using the multiply primed rolling-circle amplification (RCA) technique, which mimics the rolling-circle mechanism that occurs in nature for replication of circular DNA molecules, e.g., plasmids. The RCA protocol described here is optimized for the amplification of circular DNA virus genomes.
rAAV9--A Human-Derived Adeno-Associated Virus Vector for Efficient Transgene Expression in Mouse Cingulate Cortex: Figure 1.
Authors: Jakovcevski Mira , Guo Yin , Su Qin , Gao Guangping , Akbarian Schahram 
Published: April 2010

INTRODUCTIONThe rostro-medial cortex of the mouse and rat, considered the functional homolog to the primate prefrontal cortex (PFC), is of growing importance for preclinical models of schizophrenia and other neurodevelopmental diseases for which symptoms typically emerge in adolescence and early adulthood. Therefore, in order to explore molecular mechanisms operating during these critical stages of PFC development, it will be important to develop an efficient gene delivery system for the PFC of juvenile animals. To this end, adeno-associated virus (AAV)-based systems are increasingly used in mice for targeted gene delivery in specific brain regions such as the hippocampus. Strikingly, there is very little literature on vector-mediated gene expression in the rostro-medial cortex. In addition, multiple AAV serotypes exist based on differences in their envelope capsid proteins. However, to date, the large majority of studies in the central nervous system (CNS) have utilized the AAV2 serotype. This is typically limited to a very focal transduction pattern and therefore is not ideal for the murine PFC, which occupies several square millimeters in the rostral hemisphere. Here, we introduce a protocol for efficient, AAV9-serotype-mediated gene delivery in juvenile (postnatal day 21) and young adult PFC, resulting in long-lasting transgene expression.
Passage of pathogenic microorganisms through breathing system filters used in anaesthesia and intensive care
Authors: Scott D. H. T. , Fraser S. , Willson P. , Drummond G. B. , Baillie J. K. 
Published: April 2010

SummaryInvasive ventilation poses a risk of respiratory infection that can be drug‐resistant. One means of reducing transmission of infection is the use of a breathing system filter. Filters are intended to be used with dry gas. Current international standards do not require that filters prevent bacterial transfer when wet. It is not known whether microorganisms pass through wet filters, but theory predicts that this might occur. We tested six filters from three different manufacturers. We passed a suspension of microorganisms through the filters using the least pressure necessary, and incubated a sample of the filtrate on blood agar. All the filters tested allowed free passage of both Candida albicans and coagulase‐negative staphylococci. The median (IQR [range]) pressure required for fluid to flow across the filter varied greatly between different filter types (20 (0–48 [0–138]) cmH2O). We conclude that even large microorganisms pass across moist breathing system filters in conditions that are found in clinical practice.
Three‐dimensional seismic characterisation of large‐scale sandstone intrusions in the lower Palaeogene of the North Sea: completely injected vs. in situ remobilised sandbodies
Authors: Szarawarska Ewa , Huuse Mads , Hurst Andrew , De Boer Wytze , Lu Liwei , Molyneux Steven , Rawlinson Peter 
Published: April 2010

ABSTRACTA large number of km‐scale, saucer‐shaped sandstone bodies of enigmatic origin have recently been documented in the North Sea and the Faroe Shetland Basin. This study utilises three‐dimensional seismic data, calibrated by well data, to examine two such bodies that exhibit very similar saucer‐shaped geometries in cross‐section. The Volund and Danica structures, located 250 km apart are interpreted as end members of a spectrum of large‐scale remobilised and injected sandstones present in the North Sea Palaeogene. Both are characterised by a central 1–2 km‐wide low area surrounded by a discordant, 2–300 m tall inclined dyke complex, that tips out into a bedding concordant body interpreted as a shallow‐level sill and/or partly extruded sandstone. The origin of the central concordant sandstone body as either injected (laccolith) or depositional is of key importance to a complete understanding of the origin and prospectivity of these structures. The key criteria for recognising an injected vs. depositional origin for the central concordant sandbody are: (1) a flat, nonerosional base; (2) ‘jack‐up’ of the overburden equal to the underlying sand thickness; (3) equally thick layers of encasing mudstones; and (4) paleogeographic context. This study suggests that the Danica structure was deposited as a channel sandstone and remobilised in situ; this led to the formation of wing‐like intrusions along the channel margins. In contrast, the Volund structure overburden displays a forced‐fold geometry, arguably a diagnostic feature of an intrusive origin. The ability to recognise and differentiate completely injected vs. in situ remobilised sandbodies is important both from a basin analysis, hydrocarbon exploration and rock mechanics points of view. An improved understanding of these aspects will lead to a reduction of risks associated with the exploration and development of such a sandbody and an enhanced understanding of sediment remobilisation and fluid flow on a basin scale.
Sand‐rich injectites in the context of short‐lived and long‐lived fluid flow
Authors: Jonk Rene 
Published: April 2010

AbstractSand‐rich injectites are a common attribute of clastic sedimentary successions, and they have received increased attention in the last decade by geoscientists and engineers dealing with shallow subsurface flow and retention of aqueous and hydrocarbon fluids. Injectites form due to fluidized flow of sediment‐entraining fluids through high‐permeability strata. Given that sediment entrainment is confined to high‐permeability sediments with low horizontal effective stress and negligible cementation, the formation of sand‐rich injectites is restricted to the first kilometre of burial, where sand‐rich sediments are prone to fluidization and clay‐rich sediments are generally more cohesive and may hydraulically fracture to allow the creation of injectites. When reviewing conditions that may lead to aqueous and hydrocarbon flow within this shallow section at velocities that may cause sand fluidization and injection, we can rule out geologic time‐scale processes (disequilibrium compaction, hydrocarbon migration, and lateral pressure transfer) as plausible causes. This emphasizes the need for high‐strain‐rate processes to cause rapid fluid overpressuring and flow associated with sand injection. Earthquake‐induced shaking, instantaneous loading and release of overpressured fluids along moving fault planes are the most likely causes of sand remobilization and injection and these processes may enforce each other during tectonic stress‐release events. An additional mechanism that may trigger sand‐rich injections is the mechanical failure of shallow oil accumulations, particularly as the fluidization velocity of sand entrained in oil can be several orders of magnitude lower than for sand entrained in aqueous fluids, particularly if the oil is biodegraded (and thus has a high dynamic viscosity). Flow of hydrocarbon gas is unlikely to cause sand injection, although gas dissolved in upward‐flowing aqueous fluids may evolve at or near the surface as pressures in the aqueous phase drop.
Captivity affects immune redistribution to skin in a wild bird
Authors: Kuhlman Joshua R. , Martin Lynn B. 
Published: April 2010

Summary 1. Effects of stressors on immune functions have long been studied, but most have focused on chronic stressors, which tend to be immunosuppressive. More recently, an emphasis has been placed on identifying and understanding effects of transient, unpredictable stressors, especially whether and how they affect movement of immune cells to body areas in need of rapid protection. These latter effects have been termed, immunoredistribution. 2. In this study, we surgically implanted small gelatin sponges subcutaneously to measure leucocyte infiltration to skin in house sparrows, Passer domesticus. First, we evaluated whether wild birds exhibit leucocyte infiltration to skin in response to a transient stressor (restraint) as seen in laboratory rodents. Secondly, we examined effects of captivity on leucocyte infiltration of wild sparrows, expecting long‐duration captivity to impose chronic stress and thus reduce leucocyte infiltrations. In particular, we expected continuous elevation of the avian stress hormone, corticosterone (CORT), to dampen skin infiltration of leucocytes in birds kept in captivity for short‐term (1–2 days) and more so long‐term (1 month) periods compared with wild‐caught individuals. 3. As in lab rodents, house sparrows exhibited cutaneous leucocyte infiltration, but 1 h of restraint prior to surgery did not influence the magnitude of cellular infiltration. The leucocyte infiltration response was influenced by captivity, however, with short‐term captive birds (1–2 days) exhibiting less leucocyte influx than long‐term (30 days) and wild individuals. Interestingly, the profile of cellular infiltrates changed with time in captivity with a dramatic shift away from lymphocytes and towards heterophils in long‐term captive birds. Placement in captivity did not affect total circulating leucocytes, however, lymphocytes decreased in numbers in wild birds in short‐term captivity. CORT followed the expected pattern with the highest baseline levels in short‐term captives, intermediate levels in long‐term captives and lowest levels in wild‐caught birds. Stress responses were not affected by time in captivity. 4. These results indicate that an integral aspect of immunoredistribution occurs in wild birds but future efforts to understand the ecological and evolutionary forces that have shaped this immune response must account for the confounding effects of captivity and attempt to understand the proximate and ultimate basis for heterophilia and lymphopenia with chronic stress.
Functional analysis of the skin‐swelling response to phytohaemagglutinin
Authors: Vinkler Michal , Bainová Hana , Albrecht Tomáš 
Published: April 2010

Summary 1. The phytohaemagglutinin (PHA) skin‐swelling test is widely used in immunoecology and ecotoxicology to estimate cell‐mediated immunity. Although often presumed, the involvement of T cells in generating an immune response to PHA in vivo remains unclear. 2. To investigate the mechanism triggering this response we have compared in zebra finch (Taeniopygia guttata) the immune responses to two PHA isolectins differing in their biological properties and one control protein. 3. In Experiment I, we applied PHA‐P (the commonly used L and E isolectin mixture) into one wing‐web and bovine serum allbumin (BSA) into the other. The swelling response to PHA‐P was significantly stronger than to BSA confirming that the reaction is governed by PHA‐specific properties. 4. In Experiment II, purified PHA‐L (the T‐cell‐stimulating isolectin) and PHA‐E (the erythroagglutinating isolectin) were compared. Contrary to our expectations, PHA‐E induced a significantly stronger swelling response than PHA‐L. Histological analysis revealed significantly higher quantities of erythrocytes and thrombocytes in PHA‐E‐treated patagia. Nevertheless, there was a positive correlation between the magnitudes of these swellings. 5. In Experiment III, we tested whether the results obtained by the application of PHA‐P and pure PHA‐L differ. Here, we failed to find any significant difference between these two preparations in their immunostimulatory activity. Magnitudes of the PHA‐L‐ and PHA‐P‐induced swellings were positively correlated. 6. To the best of our knowledge this is the first study to compare the biological activity of purified PHA fractions in vivo and also the first to show the importance of erythroagglutination in the development of an inflammatory response to PHA‐P. 7. Our results indicate that the skin‐swelling test using PHA‐P reliably mirrors the individual general proinflammatory potential. However, the immunological background of the test is highly complex and the test results cannot be interpreted as measurements of the adaptive immunity or T‐cell activity. This interpretational change importantly alters our view on the test results regarding the costs of the response or the evolutionary immunological adaptations.
Trait‐mediated effect of arbuscular mycorrhiza on the competitive effect and response of a monopolistic species
Authors: Gross Nicolas , Le Bagousse‐Pinguet Yoann , Liancourt Pierre , Urcelay Carlos , Catherine Roumet , Lavorel Sandra 
Published: April 2010

Summary 1. Cessation of agricultural practices often leads to a dramatic decline in species diversity concomitant with the increase in abundance of monopolistic species, which have been hypothesized to be strongly arbuscular mycorrhiza (AM) dependent. AM can affect competitive interactions and influence plant species diversity. Few studies have tested AM effects on the competitive strategy of monopolistic species although it has been shown to be a key parameter to explain their high dominance. In addition, it is not clear whether AM effects on plant interactions can be explained by density‐mediated interactions or by trait‐mediated interactions. 2. We measured the competitive effect of Festuca paniculata, a widespread monopolistic species from subalpine communities, on itself and on two additional target species with and without AM (benomyl treatment) under contrasted fertilised treatments. AM effects on target species traits were quantified. 3. The three target species exhibited contrasted AM dependency and only F. paniculata was positively affected by AM presence in fertilised conditions. In the fertilised treatment, AM decreased intra‐specific competition and increased inter‐specific competition. Changes in competitive responses were explained by the AM effect on P inflow and species lateral spread. 4. Our results highlight AM ability to modify the performance of monopolistic species under different environmental conditions. AM provide an important mechanism by which monopolistic species can maintain a high level of dominance and dramatically decrease species diversity following agricultural abandonment.
Variation in characteristics of islets of Langerhans in insulin‐resistant, diabetic and non‐diabetic‐rat strains
Authors: Jones Huw Bowen , Nugent David , Jenkins Richard 
Published: April 2010

SummaryAssessment of the histopathological and plasma biochemical characteristics of diabetic and non‐diabetic rat strains [Han and AP Wistar, lean and obese Zucker Fatty (ZF), and lean and obese Zucker Diabetic Fatty (ZDF) rats] was performed at 6 or 14 weeks of age. Wistar and lean ZF and ZDF rats showed no or minimal islet pathology or plasma biochemical alterations at both timepoints. Obese ZFs were euglycaemic at both timepoints and mildly and severely hyperinsulinaemic at 6 and 14 weeks respectively. Islet morphology was normal at 6 weeks but at 14 weeks, islet hyperplasia was present with a minority showing degenerative changes namely, β‐cell vacuolation, vascular congestion and haemorrhage with minimal mononuclear cell and T lymphocytic infiltration. Obese ZDFs were euglycaemic and moderately hyperinsulinaemic at 6 weeks and severely hyperglycaemic with minor hypoinsulinaemia at 14 weeks. Obese ZDFs at 6 weeks showed mainly normal islets with some displaying degeneration (ranging from β‐cell vacuolation alone to the features described above). At 14 weeks, islet degeneration was more severe and widespread: β‐cell death was present in numerous islets at low level. Islet β‐cell numbers were reduced or absent (with associated reduction in insulin immunostaining) within the islets that now consisted predominantly of fibroblasts, collagen and mononuclear cells. Fibroproliferation consisting of smooth muscle actin‐α‐positive tissue was associated with mononuclear cell infiltration. Some fibrous scars were visible indicative of lost islets. Islet degeneration in obese ZF and ZDF rats was not accompanied by a reduction in β‐cell proliferation or in compensatory proliferation of β‐cell neogenic clusters. In the light of recent reports of adaptive and inflammation‐mediated degenerative changes in human non‐insulin dependent diabetes mellitus (NIDDM) islets, the hypertrophy/hyperplasia of β‐cells and islet degeneration involving infiltration by monocyte/macrophages in obese ZF and obese ZDF rats respectively offers substantial potential for elucidation of the processes involved.
Forgetfulness and older adults: concept analysis
Authors: Ballard Julianne 
Published: April 2010

ballard j. (2010) Forgetfulness and older adults: concept analysis. Journal of Advanced Nursing 66(6), 1409–1419.AbstractTitle.  Forgetfulness and older adults: concept analysis. Aim.  This paper is a report of an analysis of the concept of forgetfulness as it applies to older adults.Background.  Perceived increases in forgetfulness cause much distress for older adults due to the stigma associated with memory loss and dementia. The difference between forgetfulness in a healthy ageing state and memory loss associated with cognitive impairment requires clarification for nurses involved in screening and caring for older adults.Data sources.  Literature for this concept analysis was obtained by searches of CINAHL, EMBASE and Internurse covering the years 1962–2009, as well as gerontology textbooks.Review methods.  The evolutionary method of Rodgers was the framework for the concept analysis.Results.  As we age there is a slight increase in occasions of forgetfulness (particularly remembering to do things in the near future) that can be offset by limiting distractions and enhancing focus. This is contrasted with forgetfulness patterns, also more likely as we age, that are indicative of disease processes. Older adults’ awareness of their own forgetfulness has the potential to cause fear, embarrassment, anger and/or low self‐esteem.Conclusion.  It is important to avoid labelling or dismissal of concerns related to forgetfulness in older adults. Further research, with an emphasis on the choice of assessment tool, is recommended to investigate the benefits of nurse‐led population screening for memory deficits in older adults. Nurses require training in assessment of memory and access to adequate referral pathways if abnormal results are uncovered.
Critical thinking dispositions among newly graduated nurses
Authors: Wangensteen Sigrid , Johansson Inger S. , Björkström Monica E , Nordström Gun 
Published: April 2010

wangensteen s., johansson i.s., björkström m.e. & nordström g. (2010) Critical thinking dispositions among newly graduated nurses.Journal of Advanced Nursing 66(10), 2170–2181.AbstractAim. The aim of the study was to describe critical thinking dispositions among newly graduated nurses in Norway, and to study whether background data had any impact on critical thinking dispositions.Background. Competence in critical thinking is one of the expectations of nursing education. Critical thinkers are described as well‐informed, inquisitive, open‐minded and orderly in complex matters. Critical thinking competence has thus been designated as an outcome for judging the quality of nursing education programmes and for the development of clinical judgement. The ability to think critically is also described as reducing the research–practice gap and fostering evidence‐based nursing.Methods. A cross‐sectional descriptive study was performed. The data were collected between October 2006 and April 2007 using the California Critical Thinking Disposition Inventory. The response rate was 33% (n = 618). Pearson’s chi‐square tests were used to analyse the data.Results. Nearly 80% of the respondents reported a positive disposition towards critical thinking. The highest mean score was on the Inquisitiveness subscale and the lowest on the Truth‐seeking subscale. A statistically significant higher proportion of nurses with high critical thinking scores were found among those older than 30 years, those with university education prior to nursing education, and those working in community health care.Conclusion. Nurse leaders and nurse teachers should encourage and nurture critical thinking among newly graduated nurses and nursing students. The low Truth‐seeking scores found may be a result of traditional teaching strategies in nursing education and might indicate a need for more student‐active learning models.