RESUMEN
Previous work has demonstrated a "side-effect effect," such that intentionality is more likely to be attributed to agents who bring about negatively valenced as opposed to positively valenced side effects. The rational-scientist model explains this by suggesting that norm-violating side effects are more informative for inferring intentionality than norm-conforming side effects. In the present study we reexamined this account, addressing limitations of previous empirical tests (e.g., Uttich & Lombrozo, Cognition 116: 87-100, 2010). Side-effect valence and norm status were manipulated factorially, enabling an examination of the impact of norm status on intentionality judgments in both positively and negatively valenced side effects. Additionally, the impact of side-effect norm status on the perceived valences of side effects and agents was examined. Effects of norm status were found for both positive and negative side effects. Violation of an ostensibly neutral norm led to negative perceptions of the side effect. However, a norm status effect on intentionality judgments persisted when these effects were controlled. These results support the view that the side-effect effect is the result of the rational use of social-cognitive evidence.
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Intención , Juicio , Modelos Psicológicos , Principios Morales , Percepción Social , Adulto , Femenino , Humanos , Masculino , Adulto JovenRESUMEN
Three experiments examined how children's domain knowledge and observation of exemplars interact during concept acquisition and how exposure to novel exemplars causes revision of such knowledge. In Experiments 1 (N = 126) and 2 (N = 64), children aged 4 to 10 years were shown exemplars of fictitious animal categories that were either unrelated to, or consistent with, their prior knowledge in 25% or 75% of presented exemplars. In Experiment 3, children (N = 290) saw fictitious animal, artifact, or unfamiliar social categories that were either consistent or inconsistent with their prior knowledge in 20%, 40%, 60%, or 80% of exemplars. In the test, children made judgments about the likely co-occurence of features. In all experiments, prior knowledge and exemplar observation independently influenced children's categorization judgments. Utilization of prior knowledge was consistent across age and domain, but 10-year-olds were more sensitive to observed feature covariation. Training with larger categories increased the impact of observed feature covariation and decreased reliance on prior knowledge.
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Aprendizaje por Asociación , Formación de Concepto , Desarrollo del Lenguaje , Recuerdo Mental , Solución de Problemas , Niño , Preescolar , Femenino , Humanos , Masculino , Aprendizaje por Asociación de Pares , Psicolingüística , SemánticaRESUMEN
This unit describes the fractionation and analysis of anionic oligosaccharides and gangliosides using anion-exchange high-performance liquid chromatography (HPLC). Saccharides or gangliosides are eluted in order of the number of negative charges they possess, although the charge-to-mass ratio can also contribute to elution position.
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Cromatografía Líquida de Alta Presión/métodos , Gangliósidos/análisis , Gangliósidos/química , Oligosacáridos/análisis , Oligosacáridos/química , Animales , Cromatografía DEAE-Celulosa , Humanos , Iones/química , Factores de TiempoRESUMEN
This unit describes the fractionation and analysis of neutral oligosaccharides by high-performance liquid chromatography (HPLC) on bonded amine columns. Separation is based upon hydrogen bonding between the NH2 groups of the column and the hydroxyl groups of the oligosaccharides. A support protocol describes the reduction and desalting of neutral oligosaccharides with sodium borohydride.
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Cromatografía Líquida de Alta Presión/métodos , Oligosacáridos/análisis , Oligosacáridos/aislamiento & purificación , Aminas/química , Animales , Borohidruros/química , Iones/química , Oligosacáridos/química , Oxidación-Reducción , Sales (Química)/químicaRESUMEN
Two experiments investigated the contribution of automatic and intentional memory processes to 5- and 8-year-old children's acceptance of misinformation. Children were presented with a picture story followed by misleading postevent details that either were read to participants or were self-generated in response to semantic and perceptual hints. Children were then given a recognition test under 2 instructional conditions. In the inclusion condition children reported whether they remembered items from either of the previous phases. In the exclusion condition children were instructed to exclude postevent suggestions. Children were more likely to accept misled-generate items compared to misled-read items in the inclusion condition, but the opposite was the case under exclusion instructions. Both automaticity and recollection (cf. L. L. Jacoby, 1991) influenced misinformation acceptance, but the role of automatic processes declined with age.
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Señales (Psicología) , Memoria , Sugestión , Factores de Edad , Análisis de Varianza , Niño , Preescolar , Femenino , Humanos , Masculino , Recuerdo Mental , Comunicación PersuasivaRESUMEN
In this study the authors examine the effects of procedures adapted from the cognitive interview of R. E. Geiselman, R.P. Fisher, D.P. MacKinnon, and H.L. Holland (1985) on children's recall following exposure to misleading suggestions. Children aged 5-7 years and 9-11 years saw a videotaped story and were presented with misleading or neutral information concerning story details. All were later given free- and cued-recall tests preceded by standard interview instructions or instructions that reinstated the encoding context and encouraged exhaustive reporting. Increased recall accuracy was found following cognitive interview instructions. Both age groups were susceptible to misleading suggestions, but susceptibility was unaffected by interview type. The authors discuss the implications for interviewing child witnesses.
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Cognición , Entrevista Psicológica , Recuerdo Mental , Sugestión , Niño , Preescolar , Femenino , Humanos , MasculinoRESUMEN
The study investigated the course of developmental changes in performance on nonverbal implicit and explicit memory tests and examined the degree to which implicit memory performance is dependent upon the storage of specific perceptual information. Four-, 5-, and 10-year-old children were required to name fragmented pictures of common objects or to name and answer general knowledge questions about complete versions of the same pictures. After a 48-h retention interval, all subjects were presented with a fragmented picture identification task containing pictures identical to those present during encoding (old), pictures which were from the same basic category as the study items but which varied in their perceptual similarity to those items (same), and novel pictures which were visually and semantically unrelated to the study items (new). The amount of visual information needed to name each item (picture identification threshold) was recorded. Following identification, subjects were asked whether or not they had been shown the picture previously. All age groups showed significant priming such that the picture identification threshold for the old items was lower than that of the new pictures. A smaller but significant priming effect was obtained for the same-name items. This effect was maximized when the same-name items were perceptually similar to the study items. The magnitude of these priming effects did not vary as a function of age, but greater priming was found for those children who identified picture fragments during the study phase. In contrast, the sensitivity of recognition memory performance increased from 4 to 10 years of age. These results suggest that the processes that subserve pictorial repetition priming and recognition memory develop at different rates and that such priming is dependent upon access to specific perceptual representations of studied objects.
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Memoria/fisiología , Percepción Visual , Factores de Edad , Niño , Lenguaje Infantil , Preescolar , Femenino , Humanos , Desarrollo del Lenguaje , Masculino , Recuerdo Mental , SemánticaAsunto(s)
Acetilglucosamina/metabolismo , Proteínas del Citoesqueleto/metabolismo , Glicosiltransferasas/metabolismo , Proteínas Nucleares/metabolismo , Citoesqueleto/fisiología , Glicosilación , Fosforilación , Conformación Proteica , Procesamiento Proteico-Postraduccional , Proteínas Proto-Oncogénicas c-myc/metabolismo , Transcripción GenéticaRESUMEN
Over the past decade, a number of nuclear and cytoplasmic proteins have been identified that are modified by single N-acetylglucosamine residues attached to the hydroxyl side chain of serines or threonines (O-GlcNAc). O-GlcNAc is a dynamic modification and therefore may act in a regulatory capacity analogous to phosphorylation. To undertake site-directed mutagenesis studies of O-GlcNAc's function, it is necessary to identify the sites of glycosylation on various proteins. The current method of site mapping, which involves galactosyltransferase labeling, generation of glycopeptides by proteolysis, purification by several rounds of HPLC, and gas-phase and manual Edman sequencing, is very tedious and requires about 10 pmol of pure, labeled glycopeptide. In this report, synthetic glycopeptides were generated and used to demonstrate that O-GlcNAc-modified peptides can be rapidly identified in complex mixtures by HPLC-coupled electrospray mass spectrometry due to the partial loss of the O-linked glycan (204 amu) at a modest orifice potential. Furthermore, the exact site of glycosylation was directly identified in the low picomole range by collision-induced dissociation (CID) of the glycopeptide after removal of the O-GlcNAc by alkaline beta-elimination. The conversion of glycosylserine to 2-aminopropenoic acid (2-ap) by beta-elimination both decreased the mass of the glycopeptide by 222 amu and resulted in a CID fragment ion representing the loss of 69 amu (2-ap) instead of 87 amu (Ser) at the position of the glycosylserine. Finally, we tested this method on an identical synthetic, alpha-linked O-GalNAc-modified peptide. Like O-GlcNAc, the O-GalNAc moiety was selectively removed at a modest orifice potential; however, the beta-elimination conditions that efficiently removed the O-GlcNAc only liberated about 20% of the O-GalNAc. We conclude that the selectivity and the sensitivity of this method will make it a powerful tool for determining the sites of O-GlcNAc modification on proteins of low abundance such as transcription factors and oncogenes.
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Acetilglucosamina/análisis , Glicopéptidos/química , Oligosacáridos/química , Secuencia de Aminoácidos , Secuencia de Carbohidratos , Cromatografía Líquida de Alta Presión/métodos , Glicopéptidos/síntesis química , Glicopéptidos/aislamiento & purificación , Glicosilación , Espectrometría de Masas/métodos , Datos de Secuencia Molecular , Oligosacáridos/síntesis química , Oligosacáridos/aislamiento & purificación , Serina , Espectrometría de Masa Bombardeada por Átomos Veloces/métodos , Relación Estructura-Actividad , TreoninaRESUMEN
Class I molecules are N-linked glycoproteins encoded by the MHC. They carry cytosolic protein-derived peptides to the cell surface, displaying them to enable immune surveillance of cellular processes. Peptides are delivered to class I molecules by the transporter associated with Ag processing (TAP). Peptide association is known to occur before exposure of class I molecules to the medial Golgi-processing enzyme alpha-mannosidase II, but there is limited information regarding the location or timing of peptide binding within the earlier regions of the endoplasmic reticulum (ER)-Golgi pathway. A reported association of newly synthesized class I molecules with the ER chaperonin calnexin raises the possibility of persistence of the monoglycosylated N-linked oligosaccharide (NLO) Glc1Man8GlcNAc2, known to be recognized by this lectin. To explore these matters, we determined the structure of the NLOs on the subset of newly synthesized class I molecules awaiting the loading of peptide. We pulse-labeled murine MHC H-2Db class I molecules in RMA/S cells, which lack one of the TAP subunits, causing the great majority of the molecules to be retained for prolonged periods in an early secretory compartment, awaiting peptide binding. MHC molecules pulse-labeled with [3H]glucosamine were isolated, the NLOs specifically released and structurally analyzed by a variety of techniques. Within the chosen window of biosynthetic time, most Db molecules from parental RMA cells carried mature NLOs of the biantennary complex-type, with one to two sialic acid residues. In RMA/S cells, such chains were in the minority, the majority consisting of the precursor forms Man8GlcNAc2 and Man9GlcNAc2. No glucosylated forms were detected, nor were the later processing intermediates Man5-7GlcNAc2 or GlcNAc1Man4-5GlcNAc2. Thus, most Db molecules in TAP-deficient cells are retained in an early compartment of the secretory pathway, before the point of first access to the Golgi alpha-mannosidase I, which trims alpha 1-2 linked mannose residues, but beyond the point where the alpha 1-3-linked glucose residue is finally removed by the ER glucosidase II. Thus, structural analysis of NLOs on class I molecules within a defined biosynthetic window has established a biochemical measure of the timing of peptide association.
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Antígenos H-2/química , Oligosacáridos/química , Animales , Aniones , Transporte Biológico/inmunología , Secuencia de Carbohidratos , Línea Celular , Fraccionamiento Químico , Antígenos H-2/aislamiento & purificación , Antígenos H-2/metabolismo , Ratones , Datos de Secuencia Molecular , Oligosacáridos/aislamiento & purificación , Oligosacáridos/metabolismoRESUMEN
Regeneration of periodontal and alveolar ridge defects utilizing barrier membranes has become well established in clinical dentoalveolar reconstruction. Application of this technique has evolved from the concept of separating tissues during healing to that of providing a healing environment capable of regeneration of functional structures. The biomaterial characteristics and design of membranes employed in this technique play an important role in establishing and maintaining this environment. Barrier membranes must incorporate specific features that address the biological, mechanical, and clinical use requirements involved in regenerative treatment. Although nondegradable materials require a second surgical procedure for removal, these materials simplify certain aspects of development, production, and clinical regenerative treatment for some applications. Degradable materials introduce specific considerations and limitations regarding material selection, design, and clinical application. The progressive breakdown of degradable membranes results in dynamic changes in the mechanical and biocompatibility profiles of these materials. With present technology, these factors may limit use of degradable materials to specific applications. Membrane materials, therefore, should be selected based on a thorough understanding of the benefits and limitations inherent to the material(s) in relation to the functional requirements of specific clinical applications.
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Proceso Alveolar/fisiología , Membranas Artificiales , Periodoncio/fisiología , Regeneración , Aumento de la Cresta Alveolar , Materiales Biocompatibles , Biodegradación Ambiental , Diseño de Equipo , Regeneración Tisular Guiada Periodontal , HumanosRESUMEN
Galactosyltransferase and UDP-[3H]galactose are commonly used to identify O-linked N-acetylglucosamine (O-GlcNAc)-bearing proteins and peptides. In this report we show that immobilized Ricinus communis agglutinin I (RCA I) specifically binds in vitro galactosylated O-GlcNAc-bearing peptides, facilitating their selective isolation from complex mixtures. First, the peptide YSDSPSTST was O-GlcNAc glycosylated, galactosylated, and sialylated. Of these three glycoforms, only the one with a terminal galactose interacted with the lectin. Next, RCA I was used to isolate glycopeptides from the O-GlcNAc-bearing basic phosphoprotein (BPP) of human cytomegalovirus. BPP was overexpressed using baculovirus, [3H]galactosylated, digested with trypsin, and fractionated on RCA I. Peptides that were not galactosylated passed through the column, whereas the majority of the radiolabeled glycopeptides interacted weakly with the lectin and did not require lactose or elution. These radiolabeled peptides eluted as a broad peak with the leading edge being characterized by more hydrophobic glycopeptides and the lagging edge by less hydrophobic peptides, suggesting that the polypeptide backbone may influence the interaction with the lectin. Lactose was required to elute the remaining radiolabeled peptides, suggesting that these peptides are multiply glycosylated. The weakly interacting glycopeptides were analyzed directly by liquid chromatography/electrospray-mass spectrometry (LC/ES-MS). Glycopeptides corresponding to both of the major sites of glycosylation of BPP were identified. Thus, RCA I greatly facilitates the selective isolation of in vitro galactosylated O-GlcNAc glycopeptides from complex mixtures and substantially reduces the purification required for subsequent site-mapping by gas-phase sequencing and/or LC/ES-MS.
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Acetilglucosamina/aislamiento & purificación , Glicopéptidos/aislamiento & purificación , Acetilglucosamina/química , Aglutininas , Secuencia de Aminoácidos , Cromatografía Líquida de Alta Presión , Glicopéptidos/química , Datos de Secuencia Molecular , Plantas Tóxicas , Ricinus/químicaRESUMEN
Fluorescent tagging of free oligosaccharides by reductive amination permits sensitive detection and fractionation of these molecules. To expand the scope of this approach, we have synthesized a fluorescent reagent, 2-amino-(6-amidobiotinyl)pyridine. This reagent can tag oligosaccharides under nondegradative conditions with high efficiency. The resulting adducts show excellent fractionation by reverse-phase HPLC with sensitive detection in the low picomole range. When combined with sequential exoglycosidase digestion, stepwise sequencing of the sugar chains is possible. The biotinyl group can also be used to recover the sugar chain from reaction mixtures. The high-affinity interaction of the biotinyl group with multivalent avidin or streptavidin can be used to create the functional equivalent of neoglycoproteins carrying multiple copies of oligosaccharides of defined structure. These complexes allow the production of IgG antibodies directed against the oligosaccharide chain. They can also harness the power of (strept)avidin-biotin technology for the detection and isolation of oligosaccharide-specific receptors from native sources of recombinant libraries.
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Aminopiridinas , Biotina/análogos & derivados , Colorantes Fluorescentes , Oligosacáridos , Fosfatasa Alcalina , Aminopiridinas/síntesis química , Animales , Anticuerpos , Biotina/síntesis química , Secuencia de Carbohidratos , Cromatografía Líquida de Alta Presión , Femenino , Inmunoglobulina G/aislamiento & purificación , Espectrometría de Masas , Ratones , Ratones Endogámicos C3H/inmunología , Datos de Secuencia Molecular , Oligosacáridos/síntesis química , Oligosacáridos/química , Radioisótopos de Azufre , TritioRESUMEN
Nine and 14-year-old children with mild mental retardation and children without mental retardation matched for mental and chronological age were first shown a novel category made up of five visual figures. They were then given a recognition test with a set containing the previously presented figures, novel category members, and the category prototype. The relative contribution of prototypical and exemplar-specific information to subjects' performance on the recognition test was evaluated. Results showed that the children without mental retardation employed both forms of information in arriving at a recognition decision whereas both of the retarded groups tended to rely only on prototype information. Use of exemplar-specific information was also found to be positively correlated with IQ and, in particular, with measures of short-term memory span.
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Desarrollo Infantil , Formación de Concepto , Discapacidad Intelectual/psicología , Reconocimiento Visual de Modelos , Adolescente , Niño , Toma de Decisiones , Aprendizaje Discriminativo , Educación de las Personas con Discapacidad Intelectual , Humanos , Discapacidad Intelectual/rehabilitación , Pruebas de Inteligencia , Memoria a Corto PlazoRESUMEN
During short incubations of a Golgi apparatus-enriched subcellular fraction from rat liver with UDP-[3H]GlcNAc, label is efficiently transferred to endogenous acceptors. Most of the macromolecular radioactivity is specifically released by peptide-N4-(N-acetyl-beta-glucosaminyl)asparagine amidase, indicating that it is mainly associated with N-linked oligosaccharides. The glycoprotein acceptors are resistant to proteases unless detergent is added in amounts greater than the critical micellar concentration. This shows that the acceptors are within the lumen of intact compartments, which have the correct topological orientation expected for the Golgi apparatus in intact cells. Structural characterization of the radiolabeled N-linked oligosaccharides shows a variety of distinct neutral and anionic species. The neutral chains include bi-, tri-, and tetra-antennary molecules with terminal beta-[3H] GlcNAc residues. In vitro sialylation shows that some of the tetra-antennary chains have beta 1,3-linked Gal residues on their unlabeled antennae. An unknown modification appears to block the action of beta-galactosidase on these galactosylated oligosaccharides. Chasing the labeling reaction with a mixtures of UDP-Gal, CMP-Neu5Ac, and adenosine 3'-phosphate,5'-phosphosulfate causes an increase in the percent of radiolabeled anionic oligosaccharides. Most of the negative charge is due to sialic acid (Sia), and some appears to be in phosphodiester-linked [3H]GlcNAc. The sialylated oligosaccharides are a mixture of bi-, tri-, and tetra-antennary species with 1-3-Sia residues, and some of the [3H]GlcNAc residues are directly covered with unlabeled Gal and Sia residues. This in vitro approach should recapitulate reactions that occur in the biosynthesis of N-linked oligosaccharides in the Golgi apparatus of the intact cell. Since the conditions during labeling do not permit inter-compartmental transport, the oligosaccharides produced should represent the biosynthetic capabilities of individual Golgi compartments. Evidence is presented for a functional association of GlcNAc transferases I, II, and alpha-mannosidase II, with separation from GlcNAc transferase IV and/or V. The structures also indicate co-compartmentalization of several GlcNAc transferase(s) with beta-galactosyltransferase(s) and sialyltransferase(s). The compartmental organization of the Golgi apparatus is discussed in light of these findings.
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Aparato de Golgi/metabolismo , Hígado/metabolismo , Oligosacáridos/biosíntesis , Animales , Secuencia de Carbohidratos , Cromatografía en Gel , Cromatografía Líquida de Alta Presión , Cromatografía por Intercambio Iónico , Glicósido Hidrolasas/metabolismo , Técnicas In Vitro , Hígado/ultraestructura , Masculino , Datos de Secuencia Molecular , Octoxinol , Polietilenglicoles , Polisacáridos/metabolismo , Ratas , Ratas Sprague-Dawley , Uridina Difosfato N-AcetilglucosaminaRESUMEN
When a rat liver Golgi apparatus-enriched subcellular fraction is incubated with UDP-[3H]Gal, CMP-[3H] Neu5Ac, or [acetyl-3H]acetyl (Ac)-CoA, label is efficiently transferred to endogenous acceptors, which are resistant to added proteases, unless detergent is added at a sufficiently high concentration. Thus, the acceptors are within the lumen of intact compartments of correct topological orientation, which are likely to be similar to those of the Golgi apparatus in the intact cell. In each case, approximately 90% of the macromolecular radioactivity is specifically released by peptide-N4-(N-acetyl-beta-glucosaminyl)asparagine amidase digestion, as labeled N-linked oligosaccharides. Label from UDP-[3H]Gal is transferred to several distinct N-linked oligosaccharides, and many of these carry sialic acid (Sia) residues. This amount increases if the transfer reaction is chased with CMP-Neu5Ac. A major fraction of the [3H]Gal is directly "covered" with Sia residues, indicating that at least a portion of the beta-galactosyltransferase(s) are co-localized with one or more sialyltransferases. The majority of the [3H]Gal is found in a beta 1,3-linkage, rather than the more common beta 1,4-linkage. The N-linked oligosaccharides labeled by CMP-[3H] Neu5Ac carry labeled Sia residues in either alpha 2,3 or alpha 2,6 linkage, and showed a range of charge distribution. The transferred [3H]Neu5Ac is not O-acetylated even when Ac-CoA is added at saturating concentrations, implying that the sialyltransferases and the O-acetyltransferase(s) are not functionally co-localized. However, approximately 20% of label released from N-linked oligosaccharides by sialidase does not co-migrate with authentic Neu5Ac in high performance liquid chromatography analysis, indicating that transferred [3H] Neu5Ac is modified by unknown enzymes in the Golgi. Most of the [3H]acetate transferred from [acetyl-3H] Ac-CoA to N-linked oligosaccharides is on Sia residues that are exclusively alpha 2,6-linked, and is enriched on tri- and tetra-antennary chains that do not appear to carry any 2,3-linked Sia residues. These data indicate a restricted substrate preference of the O-acetyltransferase(s). About one-quarter of the [3H]acetate transferred is sialidase-resistant, indicating either transfer to monosaccharides other than sialic acid, or to sialidase-resistant sialic acids. While most of these sialidase-resistant oligosaccharides remain negatively charged, about 10% are neutralized by sialidase, confirming transfer of [3H]acetate to monosaccharides other than sialic acid.
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Aparato de Golgi/metabolismo , Hígado/metabolismo , Oligosacáridos/biosíntesis , Acetilcoenzima A/metabolismo , Animales , Secuencia de Carbohidratos , Cromatografía de Afinidad , Cromatografía en Gel , Cromatografía Líquida de Alta Presión , Ácido N-Acetilneuramínico Citidina Monofosfato , Glicósido Hidrolasas/metabolismo , Técnicas In Vitro , Hígado/ultraestructura , Masculino , Datos de Secuencia Molecular , Polisacáridos/metabolismo , Ratas , Ratas Sprague-Dawley , Tritio , Uridina Difosfato GalactosaRESUMEN
Endogenous acceptors in a Golgi apparatus-enriched subcellular fraction from rat liver were labeled with UDP-[3H]GalNAc. The great majority of these acceptors were protected from protease degradation in the absence of detergent. These molecules are therefore present in intact vesicles of the correct topological orientation, which are likely to be similar to the Golgi compartments of the intact cell. Several distinct glycoproteins are labeled, but most are different from those labeled with UDP-[3H]GlcNAc. The enzyme peptide-N4(N-acetyl-beta-glucosiminyl)asparagine amidase releases label from a few specific proteins, indicating that [3H]GalNAc is transferred to N-linked oligosaccharides. Both neutral and anionic N-linked oligosaccharides are found, the great majority of which do not bind to ConA-Sepharose. Most of the [3H]GalNAc found in neutral oligosaccharides is terminal and beta-linked. The negative charge on the anionic molecules is due to sialic acid, and phosphate. A major portion of the [3H] GalNAc in this fraction is acid labile, and is released with kinetics consistent with it being in a phosphodiester linkage. These results show the existence of a whole new class of GalNAc-containing N-linked oligosaccharides, and demonstrates that this in vitro approach can detect previously undescribed structures. O-linked oligosaccharide biosynthesis was also studied in the same labeled rat liver Golgi apparatus preparations. beta-Elimination releases approximately 95% of the peptide-N4-(N-acetyl-beta-glucosaminyl)asparagine amidase (PNGase F)-resistant label which, in the absence of other added nucleotides, is almost exclusively [3H] GalNAcitol. If other unlabeled sugar nucleotides and adenosine 3'-phosphate,5'-phosphosulfate are added during the chase period two anionic O-linked oligosaccharides are synthesized, indicating that the UDP-GalNAc:peptide-N-acetylgalactosaminyltransferase is at least in part functionally co-localized with enzymes that extend and modify O-linked oligosaccharides.
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Hígado/metabolismo , Oligosacáridos/biosíntesis , Animales , Secuencia de Carbohidratos , Cromatografía Líquida de Alta Presión , Cromatografía en Papel , Electroforesis en Gel de Poliacrilamida , Concentración de Iones de Hidrógeno , Técnicas In Vitro , Cinética , Hígado/ultraestructura , Datos de Secuencia Molecular , Polisacáridos/metabolismo , Ratas , Uridina Difosfato N-AcetilgalactosaminaRESUMEN
This study is an attempt to clarify the nature of developmental differences in the use of prototypical features or information about specific exemplars for object categorization and to identify stimulus factors that may modulate the use of these information sources. To this end, 6-year-olds, 11-year-olds, and adults were taught to sort visual patterns into one of two overlapping categories. Immediately or 24 h following category acquisition subjects were presented with a set of transfer stimuli consisting of both old and new patterns, including the theoretical prototype. Categorization responses to these test stimuli were examined against predictions derived from the prototype and nearest-old-exemplar accounts of categorization. A significant developmental change in the process of categorization was noted. For the youngest group, only the prototype model was found to fit the test data, whereas for the older groups both models independently explained significant amounts of variance in performance. This trend was not affected by the delay between training and testing, nor by a manipulation of intracategory variability. The emergence of the specific exemplar model as a viable explanation of ill-defined categorization thus appears to be related to the developmental level attained.
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Atención , Desarrollo Infantil , Aprendizaje Discriminativo , Reconocimiento Visual de Modelos , Adulto , Niño , Femenino , Humanos , Masculino , Orientación , Solución de ProblemasRESUMEN
Tritiated uridine-5'-diphosphogalactose (UDP-[3H]Gal) has been widely used to study oligosaccharide biosynthesis and structure. It can be synthesized either chemically or enzymatically using galactose oxidase to oxidize the hydroxyl moiety at C-6 to an aldehyde (6-aldo-UDP-Gal), which is then reduced back to the alcohol with tritiated sodium borohydride. Although the enzymatic approach is simple and efficient, there are several problems associated with it. First, incomplete oxidation to the aldehyde reduces the final specific activity. Second, if the galactose oxidase is not removed from the 6-aldo-UDP-Gal prior to reduction, the resulting UDP-[6-3H]Gal can be reoxidized to 6-aldo-UDP-[6-3H]Gal. We present evidence for the occurrence of this compound in one commercially obtained preparation of UDP-[6-3H]Gal. Finally, if an excess of 6-aldo-UDP-Gal is used for good yield, it is necessary to quench the reduction with nonradioactive borohydride, again reducing the final specific activity. We have devised a rapid, inexpensive, and efficient synthesis of UDP-[6-3H]Gal that circumvents all of these problems. Galactose oxidase is used to produce 6-aldo-UDP-Gal and the completeness of this reaction is confirmed on polyethyleneimine (PEI) cellulose TLC plates. The 6-aldo-UDP-Gal is purified on silica gel 60 TLC plates. This purified compound is then reduced with tritiated sodium borohydride, with the aldehyde present in excess. Unreacted 6-aldo-UDP-Gal is then purified away from the product UDP-[6-3H]Gal by chromatography on PEI cellulose. Radiochemically pure UDP-[6-3H]Gal with a specific activity of 10 Ci/mmol was obtained using the above scheme.(ABSTRACT TRUNCATED AT 250 WORDS)