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Thinking in nursing education. Part II. A teacher's experience.

Across academia, educators are investigating teaching strategies that facilitate students' abilities to think critically. Because may these strategies require low teacher-student ratios or sustained involvement over time, efforts to implement them are often constrained by diminishing resources for education, faculty reductions, and increasing number of part-time teachers and students. In nursing, the challenges of teaching and learning critical thinking are compounded by the demands of providing care to patients with increasingly acute and complex problems in a wide variety of settings. To meet these challenges, nurse teachers have commonly used a variety of strategies to teach critical thinking (1). For instance, they often provide students with case studies or simulated clinical situations in classroom and laboratory settings (2). At other times, students are taught a process of critical thinking and given structured clinical assignments, such as care plans or care maps, where they apply this process in anticipating the care a particular patient will require. Accompanying students onto clinical units, teachers typically evaluate critical thinking ability by reviewing a student's preparation prior to the experience and discussing it with the student during the course of the experience. The rationales students provide for particular nursing interventions are taken as evidence of their critical thinking ability. While this approach is commonly thought to be effective, the evolving health care system has placed increased emphasis on community nursing (3,4), where it is often difficult to prespecify learning experiences or to anticipate patient care needs. In addition, teachers are often not able to accompany each student to the clinical site. Thus, the traditional strategies for teaching and learning critical thinking common to hospital-based clinical courses are being challenged, transformed, and extended (5). Part II of this article describes findings that suggest how many teachers and students are challenging the conventional approaches to schooling and creating pedagogies that are more responsive to the contemporary context of health care.

Attitude of Health Personnel↗

Biasing the brain's attentional set: II. effects of selective intersensory attentional deployments on subsequent sensory processing.

This study used high-density mapping of human event-related potentials to examine the brain activity associated with selective information processing when subjects were cued on a trial-by-trial basis to perform a discrimination in either the visual or auditory modality. On each trial, word-cues (S1) instructed subjects to attend to features within one sensory-modality of an impending compound auditory-visual stimulus (S2) that arrived approximately 1-second following the cue. Subjects made a discrimination within the cued modality of the S2 stimulus. The spatio-temporal patterns of activity in response to the compound S2 stimulus were examined as a function of the sensory modality being attended. The earliest effects of intersensory attention on visual processing were seen subsequent to the initial activation of visual cortex, beginning at 80 ms and continuing into the P1 and N1 components of the visual ERP. The scalp-topography of this earliest modulation was consistent with modulation of activity in ventral visual stream areas. Thus, the locus of effects on visual S2 processing differed from the anticipatory parieto-occipital biasing activity that preceded S2 presentation. This pattern of effects strongly suggests that the anticipatory activity (following the cue) associated with sustaining the focus of attention during intersensory attention, at least in the context of this paradigm, does not operate as a simple gain mechanism in early visual sensory areas. Rather, attentional biasing can operate through a higher-order process whereby parieto-occipital cortices influence the subsequent flow of visual processing in the ventral stream.

Acoustic Stimulation↗

A dominant B-cell epitope on the 22 kDa tegumental membrane-associated antigen of Schistosoma japonicum maps to an EF-hand calcium binding domain.

The 22 kDA tegumental surface membrane-associated antigen of schistosomes is of recognized importance in immunity to schistosomiasis. Here, we have defined linear B-cell epitopes on the recently cloned and expressed recombinant Schistosoma japonicum 22 kDa antigen (reSj22), using mice immunized with this molecule. Sera from three strains of mice, CBA, C57BL/6 and BALB/c, representing different genetic backgrounds, were reacted with a series of overlapping synthetic peptides in epitope-scanning studies. The predicted hydrophilic N-terminal domain was found to be highly immunogenic, containing several sequences that were strongly recognized by all strains of mice. The most dominant epitope identified, corresponding to peptide 6, was located in a region previously identified as an EF-hand calcium binding domain. In contrast, the more hydrophobic C-terminal region of the molecule was poorly recognized, with the exception of a single peptide encoding residues 121-140 that was recognized by CBA and C57BL/6 but not BALB/c mice, suggesting that the latter epitope was genetically restricted between the strains. The data presented here describing the epitope mapping of this molecule may prove important in research aimed at further defining immune responses to schistosomal antigens.

Amino Acid Sequence↗

Spatial analysis of evoked potentials in man--a review.

Steps in brain information processing are reflected on the scalp as changes of the electric potential which is evoked by the stimulus. However, for a given recording point on the scalp, there is no absolute amplitude or phase information of the electric brain potential. This means that the shape of an evoked potential waveform which is recorded from a given scalp location crucially depends on the location of the chosen reference. Only unbiased results of evoked potential data evaluation can be hoped to elucidate or map successfully into information processing models established by other methods, e.g. behavior measurements. Conventional recordings vs a common reference contain only one of many possible sets of waveshapes. In order to avoid ambiguities or bias of results, the entire evoked potential data set firstly must be analysed over space, and reference-independent parameters must be extracted. For each time point, the spatial distribution of the potentials is viewed as field map. The parameter extraction in a direct approach at each time point includes, e.g. locations of field peaks and troughs, voltage and gradient between them, and global electrical field power; or, parameters via the first or second spatial derivative of the electric field. In the second step, changes of these reference-independent field measurements are analysed over time. At component latency which is defined by maximal, global field power or by voltage range, mapped field distributions can be compared using maximal/minimal field value locations or complete maps. Significantly different field configurations establish the activity of non-identical neural generators. Classification of the field configurations (examination of orbits of field extrema over time) leads to the segmentation of series of field maps (multichannel EP data) into short epochs of stationary spatial configurations (i.e. spatially characterized components) with equal consideration of all recording points, and without the amplitude criterion. The application of these principles to the following problems is discussed: comparison of evoked potentials between different analysis times, in particular pre-stimulus and post-stimulus electric brain states; zero baseline for measurement; reference electrode; identification of evoked components in time and space. Illustrations of these problems include functional differences of input-analysing sub-systems, and the topography of cognition- and speech-related electric brain activity.

Brain↗

Dual representation of the hand in the cerebellum: activation with voluntary and passive finger movement.

Early electrophysiological studies during sensory stimulation in the anesthetized cat and more recent functional imaging studies during voluntary movement in humans have provided evidence for two separate representations of the body in the anterior and posterior lobes of the cerebellum; however, the functional role of these body maps in motor and sensory processing is not known. The aims of the present study were to determine whether this dual representation is also present during passive movement, and to compare the pattern of activation with that obtained during voluntary movement. Functional MRI measurements were undertaken in 14 subjects who performed right index finger flexion and extension movements at approximately 1 Hz, or had their finger moved passively at the same rate and through the same angle using a pneumatic device. During passive movement, dual activation was detected in the ipsilateral cerebellum, in the anterior lobe, and in the posterior lobe. A similar pattern of activation was observed during voluntary movement; however, the overall magnitude was about doubled. These data provide evidence for a dual ipsilateral representation of the hand in the rostral and caudal cerebellar cortex during passive as well as voluntary movements, with the rostral representation being the dominant one, and indicate that both of these areas are involved in kinesthetic sensory and motor processing.

Adult↗

Further argument for the existence of a pacemaker in the human information processing system.

To support the idea that temporal information processing may depend on an internal clock, Treisman et al. proposed a pacemaker model (Treisman, M., Faulkner, A., Naish, P.L.N., Brogan, D., 1990. The internal clock: Evidence for a temporal oscillator underlying time perception with some estimates of its characteristics frequency. Perception 19, 705-743.) and a technique for interfering with it by introducing an external periodic phenomenon. Experimental results obtained by these authors on time estimation and production tasks support this model. In another study, Treisman et al. established that the pacemaker also affects reaction times (RT) (Treisman, M., Faulkner, A., Naish, P.L.N., 1992. On the relation between time perception and the timing of motor action: Evidence for a temporal oscillator controlling the timing of movement. Quarterly Journal of Experimental Psychology 45A, 235-263.). In the present study, we addressed the question as to which information processing stage (Sanders, A.F., 1980. Stage analysis of reaction process, In: Stelmach, G.E., Requin, J. (Eds.). Tutorials in motor behavior. North-Holland, Amsterdam, pp. 331-354.) is affected by this internal clock. For this purpose, we used the Additive Factors Method (Sternberg, S., 1969. The discovery of processing stages: Extension of Donder's method. In: Koster, W.G. (Ed.). Attention and Performance II. Acta Psychologica 30, 276-315.). To vary sensorial processing time, we used two visual stimulus intensities. Stimulus-response mapping was manipulated to enhance central processing time. To modify the duration of the motor stages, the two responses could be given by two fingers on the same hand (right ring vs. middle finger) or by two fingers of the different hands (right ring vs. left middle finger). Intensity of the stimulus, stimulus-response mapping, and repertoire of responses were found to be additive. We obtained RT modulations similar to those obtained by Treisman et al. in 1992. No first order interactions were observed between the periodical phenomenon and the other manipulated factors but only a third order one. Two possible interpretations of these results are proposed.

Adult↗

Three-Dimensional Representations of Contour Maps

Contour map learning may require three-dimensional representation of the area depicted by a contour map. The purpose of the study was to test the hypothesis that participants created three-dimensional mental representations of contour maps when asked to generate a cross-section profile of the map terrain from one point on the contour map to another. Gender differences in cross-section performance were also investigated, and it was hypothesized that males would perform better than females in a contour map cross-section test. Participants studied a series of contour maps or landsurface maps (three-dimensional drawings of an area), and answered two cross-section questions per map. Following the cross-section test, participants were given an incidental recognition test for the previously studied maps, either in a contour map format or a landsurface map format. Males recognized the landsurface maps corresponding to the contour map cross-section questions answered correctly better than contour map cross-section questions answered incorrectly, whereas females did not. This finding suggested that males, but not females, formed three-dimensional representations of the contour maps. There were no gender differences in performance on the contour map cross-section test, but males achieved higher recognition scores than females when the cross-section stimuli were contour maps. It was concluded that multiple spatial and verbal processing strategies can be used successfully to solve a contour map cross-section test, but that three-dimensional spatial processing may be more efficient than other forms of processing for long-term memory of contour maps. Copyright 1998 Academic Press.

Journal Article↗

Genome-wide localization of histone 4 arginine 3 methylation in a differentiation primed myeloid leukemia cell line.

Methylation of arginine residues in proteins is involved in modulation of various protein-protein interactions. At the chromatin level H4R3 methylation provides a signal integration step during myeloid differentiation. In order to globally characterize the role of arginine methylation in signal integration and developmental processes we decided to map genomic loci marked by protein arginine methyl transferase 1 (PRMT1) via histone H4 arginine 3 methylation. For this, we used the myeloid leukemia cell line, HL60, which is known to differentiate along the monocyte/macrophage or granulocyte lineage. We used chromatin immunoprecipitation with an antibody specific for the H4 arginine 3 methyl epitope followed by cloning to isolate genomic loci marked by this modification. After sequencing and in silico analysis we found that all of the genomic hits identified were intronic or within 5 kb of 5' ends of specific genes. The locations identified were enriched in conserved transcription factor binding sites of POU2F1, MEF-2 and FOXL1 factors. A significant number of the genes in the proximity of the identified genomic loci are involved in signaling pathways and developmental processes including immune response of myeloid cells.

Arginine↗

Comparison of experimental and computational functional group mapping of an RNase A structure: implications for computer-aided drug design.

One relatively new computational approach to the drug discovery process involves calculating functional group maps of a target structure. Experimental functional group mapping techniques have also recently emerged. In this paper, the structure of RNase A with two bound formates (i.e. carboxylate functionalities) is used as a model system to test the computational methodology. Functional group maps of the RNase A structure were calculated using the Multiple Copy Simultaneous Search (MCSS) method and compared with experimentally determined formate and water positions. The calculations indicate that the protonation state of active-site histidines determines the ability of the enzyme to bind formate. The results also suggest an ordered binding mechanism for the two formates. An improved strategy for using the MCSS method to design new candidate ligands is discussed.

Animals↗

Characterization of aggrecan retained and lost from the extracellular matrix of articular cartilage. Involvement of carboxyl-terminal processing in the catabolism of aggrecan.

The catabolism of aggrecan in bovine articular cartilage explants is characterized by the release into the culture medium of high molecular weight aggrecan fragments, generated by the proteolytic cleavage of the core protein between residues Glu373 and Ala374 within the interglobular domain. In this study, the position of the carboxyl-terminus of these aggrecan fragments, as well as a major proteolytically shortened aggrecan core protein present in cartilage matrix, have been deduced by characterizing the peptides generated by the reaction of aggrecan core protein peptides with cyanogen bromide. It was shown that two out of three such peptide fragments having an amino terminus starting at Ala374 have their carboxyl terminus located within the chondroitin sulfate 1 domain. The third and largest aggrecan core protein peptide, with an amino terminus starting at Ala374, has a carboxyl terminus in a region of core protein between the chondroitin sulfate 1 domain and the chondroitin sulfate 2 domain. The carboxyl terminus of this peptide appeared to be the same as that of the proteolytically degraded aggrecan core protein, which is retained within the extracellular matrix of the tissue. Another two aggrecan fragments recovered from the medium of explant cultures with amino-terminal sequences in the chondroitin sulfate 2 domain at Ala1772 and Leu1872 were shown to have their carboxyl termini within the G3 globular domain. These results suggest that the catabolism of aggrecan between residues Glu373 and Ala374 in the interglobular domain by the putative proteinase, aggrecanase, may be dependent on prior proteolytic processing within the carboxyl-terminal region of the core protein.

Aggrecans↗

Mapping domain structures in silks from insects and spiders related to protein assembly.

The exceptional solubility in vivo (20-30%, w/v) of the silk proteins of insects and spiders is dictated by both the need to produce solid fibres with a high packing fraction and the high mesogen concentration required for lyotropic liquid crystalline spinning. A further design requirement for silk proteins is a strong predominance of hydrophobic amino acid residues to provide for the hydrophobic interactions, water exclusion, and beta-crystallite formation required to produce strong insoluble threads. Thus, the domain structure of silk proteins needs to enable nanoscale phase separation to achieve high solubility of hydrophobic proteins in aqueous solutions. Additionally, silk proteins need to avoid premature precipitation as beta-sheets during storage and processing. Here we use mapping of domain types, sizes and distributions in silks to identify consistent design features that have evolved to meet these requirements. We show that silk proteins consist of conspicuously hydrophilic terminal domains flanking a very long central portion constructed from hydrophobic blocks separated by hydrophilic ones, discussing the domain structure in detail. The general rules of construction for silk proteins based on our observations should give a useful guide to the way in which Nature has solved the problem of processing hydrophobic proteins in water and how this can be copied industrially. Following these rules may also help in obtaining adequate expression, soluble products and controllable conformational switches in the production of genetically engineered or chemically synthesized silk analogues. Thus these insights have implications for structural biology and relevance to fundamental and applied questions in material science and engineering.

Animals↗

Chemical and photo-oxidative hair damage studied by dye diffusion and electrophoresis.

Microspectrophotometric and electrophoretic methods were used to characterize and quantify the effects of primary damage to hair from chemical and photochemical oxidative processes. The diffusion of molecules proceeding from the fiber surface to the center of untreated and modified (by chemical and photochemical oxidative processes) hair fibers was mapped by fluorescence microscopy and quantified by calculating diffusion coefficients of a fluorescent molecule. In addition, an electrophoretic separation technique, namely, SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis), was used not only to substantiate the results obtained in the microfluorometric study, but also to show how the main classes of proteins of unaltered hair are modified by cosmetic chemical treatments, light exposure, and combinations of these two processes. UV microspectrophotometry is an alternate analytical method to evaluate photo-oxidative damage in hair, and supports the results obtained by microfluorometry.

Diffusion↗

Protein interaction mapping for target validation: the need for an integrated combinatory process involving complementary approaches.

Identification, selection, validation and prioritization of targets for therapeutic intervention requires understanding of the biological role of individual proteins in cellular pathways. Unraveling the ways in which proteins interact with each other appears to be crucial in achieving that goal. A number of recently described high-throughput approaches for analyzing cellular protein-protein interactions and previously proposed prediction procedures are compared in this review. The relative advantages of each method are discussed in relation to reproducibility, comprehensiveness and biological significance. It is concluded that only a combination of complementary biochemical technologies supported by reliable algorithms, will provide exhaustive maps of protein interactions for a cellular interactome.

Animals↗

Frequent emergence and functional resurrection of processed pseudogenes in the human and mouse genomes.

Despite the wide distribution of processed pseudogenes in mammalian genomes, such as those of human and mouse, relatively little is known about their roles in genomic evolution. While gene duplications are recognized as one of the major driving forces in genome evolution, processed pseudogenes, which are retrotransposed copies of mRNAs, have been regarded as junk or selfish DNA for a long time. In order to elucidate the quantitative and qualitative contribution of processed pseudogenes to the mammalian genome evolution, we attempted to detect processed pseudogenes by extensively mapping the mRNAs to both the human and mouse genomes, and then we estimated the rate of their emergence. As a result, we revealed that the rate of pseudogene emergence was about 1-2% per gene per million years, which was as high as the rate (0.9%) of gene duplication in the human genome, although the rate of pseudogene emergence was found to drastically decrease in the hominid lineage. Furthermore, 1% of the processed pseudogenes seemed to be reinvigorated by post-retrotransposition transcription, many of them preserving the intact coding regions. Since the expression patterns of transcribed pseudogenes in various tissues were quite different between human and mouse, their emergence might have led to species-specific evolution. Our results indicate that the generation of processed pseudogenes was not wholly futile but instead has been an indispensable resource, driving dynamic evolution of the mammalian genomes.

Amino Acid Sequence↗

Phosphorelay-regulated degradation of the yeast Ssk1p response regulator by the ubiquitin-proteasome system.

In Saccharomyces cerevisiae, a phosphorelay signal transduction pathway composed of Sln1p, Ypd1p, and Ssk1p, which are homologous to bacterial two-component signal transducers, is involved in the osmosensing mechanism. In response to high osmolarity, the phosphorelay system is inactivated and Ssk1p remains unphosphorylated. Unphosphorylated Ssk1p binds to and activates the Ssk2p mitogen-activated protein (MAP) kinase kinase kinase, which in turn activates the downstream components of the high-osmolarity glycerol response (HOG) MAP kinase cascade. Here, we report a novel inactivation mechanism for Ssk1p involving degradation by the ubiquitin-proteasome system. Degradation is regulated by the phosphotransfer from Ypd1p to Ssk1p, insofar as unphosphorylated Ssk1p is degraded more rapidly than phosphorylated Ssk1p. Ubc7p/Qri8p, an endoplasmic reticulum-associated ubiquitin-conjugating enzyme, is involved in the phosphorelay-regulated degradation of Ssk1p. In ubc7Delta cells in which the degradation is hampered, the dephosphorylation and/or inactivation process of the Hog1p MAP kinase is delayed compared with wild-type cells after the hyperosmotic treatment. Our results indicate that unphosphorylated Ssk1p is selectively degraded by the Ubc7p-dependent ubiquitin-proteasome system and that this mechanism downregulates the HOG pathway after the completion of the osmotic adaptation.

Biological Transport↗

Structure of a major yolk glycoprotein and its processing pathway by limited proteolysis are conserved in echinoids.

To study the fate of the yolk glycoproteins found in eggs and embryos of the sea urchin, Strongylocentrotus purpuratus, a polyclonal antibody to a 90-kDa polymannose glycoprotein found in the embryo was prepared. Immunoblot analysis of total proteins over the course of development showed that this antibody recognized a family of glycoproteins. Concomitant with the disappearance of the major 160-kDa yolk glycoprotein of the egg during embryogenesis, glycoproteins with a lower molecular mass appeared. These glycoproteins (115, 108, 90, 83, and 68 kDa) were purified from S. purpuratus and analyzed by limited proteolysis and peptide mapping. This analysis revealed that these glycoproteins were cleavage products derived from the major yolk glycoprotein. The antibody to the 90-kDa glycoprotein in S. purpuratus embryos was used to identify a homologous set of yolk glycoproteins with similar molecular masses in the embryos of three other species in the class Echinoidea: Arbacia punctulata, Lytechinus pictus, and Dendraster excentricus. However, eggs from other echinoderm classes and from Xenopus laevis, Drosophila melanogaster, and the chicken did not contain any cross-reactive molecules. Cross-reactivity within the class Echinoidea was not due to a common carbohydrate epitope, because the antibody recognized the glycoproteins even after the N-linked carbohydrate side chains were enzymatically removed. The major yolk glycoprotein (160-170 kDa) from each of the three sea urchin species was purified and analyzed. Comparison of the physical and chemical properties of these glycoproteins revealed striking similarities in pI and in amino acid and monosaccharide composition. The results of peptide mapping also supported the conclusion that the 160- to 170-kDa glycoproteins from the four echinoids are structurally homologous glycoproteins containing N-linked polymannose chains. Immunolocalization by electron microscopy in S. purpuratus showed that the yolk glycoproteins remained within the yolk platelet throughout development, and that externalization of the 160-kDa glycoprotein or its cleavage products was not detectable.

Amino Acids↗

Two-dimensional crystallization of a membrane protein on a detergent-resistant lipid monolayer.

Two-dimensional crystals of a membrane protein, the proton ATPase from plant plasma membranes, have been obtained by a new strategy based on the use of functionalized, fluorinated lipids spread at the air-water interface. Monolayers of the fluorinated lipids are stable even in the presence of high concentrations of various detergents as was established by ellipsometry measurements. A nickel functionalized fluorinated lipid was spread into a monolayer at the air-water interface. The overexpressed His-tagged ATPase solubilized by detergents was added to the subphase. 2D crystals of the membrane protein, embedded in a lipid bilayer, formed as the detergent was removed by adsorption. Electron microscopy indicated that the 2D crystals were single layers with dimensions of 10 microm or more. Image processing yielded a projection map at 9 A resolution, showing three well-separated domains of the membrane-embedded proton ATPase.

Adsorption↗