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Chromothripsis is a catastrophic cellular event recently described in cancer in which chromosomes undergo massive deletion and rearrangement. Here, we report a case in which chromothripsis spontaneously cured a patient with WHIM syndrome, an autosomal dominant combined immunodeficiency disease caused by gain-of-function mutation of the chemokine receptor CXCR4. In this patient, deletion of the disease allele, CXCR4(R334X), as well as 163 other genes from one copy of chromosome 2 occurred in a hematopoietic stem cell (HSC) that repopulated the myeloid but not the lymphoid lineage. In competitive mouse bone marrow (BM) transplantation experiments, Cxcr4 haploinsufficiency was sufficient to confer a strong long-term engraftment advantage of donor BM over BM from either wild-type or WHIM syndrome model mice, suggesting a potential mechanism for the patient's cure. Our findings suggest that partial inactivation of CXCR4 may have general utility as a strategy to promote HSC engraftment in transplantation.
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Inestabilidad Cromosómica , Síndromes de Inmunodeficiencia/genética , Verrugas/genética , Animales , Cromosomas Humanos , Modelos Animales de Enfermedad , Haploinsuficiencia , Células Madre Hematopoyéticas/metabolismo , Humanos , Linfocitos/metabolismo , Masculino , Ratones , Persona de Mediana Edad , Mosaicismo , Mutación , Células Mieloides/metabolismo , Enfermedades de Inmunodeficiencia Primaria , Receptores CXCR4/genética , Remisión EspontáneaRESUMEN
People accused of sexual assault are often described as the "real" victim by their defenders, but the impact of "victim framing" on public opinion is unknown. We investigated this issue across four experiments (N = 2,614). Online U.S. adult participants read a report about an alleged sexual assault that framed the female accuser as the victim (of assault), framed the male alleged perpetrator as the victim (of false accusations), or was neutral about victimhood (baseline). Relative to those in the baseline condition, participants in the assault- and allegation-victim conditions generally expressed more support for the victim-framed protagonist and less support for the other protagonist. The consistency of these effects varied with how often the victim frame was instantiated and whether the report described a fictionalized or real-world case. Across all contexts, however, participants who identified the victim-related language as influencing their evaluations exhibited strong framing effects. This suggests that social-pragmatic reasoning is a key mechanism by which victim framing shapes moral judgments.
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Víctimas de Crimen , Delitos Sexuales , Adulto , Actitud , Femenino , Humanos , Juicio , MasculinoRESUMEN
Bermúdez persuasively argues that framing effects are not as irrational as commonly supposed. In focusing on the reasoning of individual decision-makers in complex situations, however, he neglects the crucial role of the social-communicative context for eliciting certain framing effects. We contend that many framing effects are best explained in terms of basic, rational principles of discourse processing and pragmatic reasoning.
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Conducta de Elección , Toma de Decisiones , Masculino , Humanos , Solución de ProblemasRESUMEN
Mucopolysaccharidosis III B (MPS III-B) is a rare lysosomal storage disorder caused by deficiencies in Alpha-N-acetylglucosaminidase (NAGLU) for which there is currently no cure, and present treatment is largely supportive. Understanding the structure of NAGLU may allow for identification of novel therapeutic targets for MPS III-B. Here we describe the first crystal structure of human NAGLU, determined to a resolution of 2.3â¯Å. The crystal structure reveals a novel homotrimeric configuration, maintained primarily by hydrophobic and electrostatic interactions via domain II of three contiguous domains from the N- to C-terminus. The active site cleft is located between domains II and III. Catalytic glutamate residues, E316 and E446, are located at the top of the (α/ß)8 barrel structure in domain II. We utilized the three-dimensional structure of NAGLU to map several MPS III-B mutations, and hypothesize their functional consequences. Revealing atomic level structural information about this critical lysosomal enzyme paves the way for the design of novel therapeutics to target the underlying causes of MPS III-B.
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Acetilglucosamina/química , Acetilglucosaminidasa/química , Acetilglucosamina/metabolismo , Acetilglucosaminidasa/genética , Acetilglucosaminidasa/metabolismo , Secuencias de Aminoácidos , Dominio Catalítico , Línea Celular Tumoral , Clonación Molecular , Cristalografía por Rayos X , Fibroblastos/citología , Fibroblastos/metabolismo , Expresión Génica , Vectores Genéticos/química , Vectores Genéticos/metabolismo , Humanos , Interacciones Hidrofóbicas e Hidrofílicas , Modelos Moleculares , Mucopolisacaridosis III/enzimología , Mucopolisacaridosis III/genética , Mucopolisacaridosis III/patología , Mutación , Unión Proteica , Conformación Proteica en Hélice alfa , Conformación Proteica en Lámina beta , Dominios y Motivos de Interacción de Proteínas , Multimerización de Proteína , Estructura Terciaria de Proteína , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Electricidad Estática , Homología Estructural de Proteína , Especificidad por SustratoRESUMEN
Today's state-of-the-art cell sorting flow cytometers are equipped with aerosol containment systems designed to evacuate aerosols from the sort chamber during a sort. This biosafety device is especially important when the sort operator is sorting infectious or potentially infections samples. Hence, it is critical to evaluate the performance for this system in normal operation and in "failure" mode to determine the efficacy of containment. In the past decade, the most popular published method for evaluating containment has been the Glo-Germ bead procedure. These highly fluorescent and multisize particles can easily be detected on a microscope slide and enumerated using a fluorescent microscope. Collecting particles on this slide is accomplished using an Aerotech impactor. This sampler collects potentially escaping aerosols from the sort chamber before enumerating any particles. Although the Glo-Germ procedure has been adopted by many labs, there are several drawbacks with the procedure that have limited its adoption by cell sorter laboratories: The Aerotech impactor is a reusable device that requires rigorous cleaning between measurements. The surface area of the collection slide is large and difficult to scan on a fluorescence microscope. These beads produce a wide variation in sizes resulting in inconsistency in flow rates. Here, we describe a novel and replacement method utilizing a Cyclex-d impactor and Dragon Green beads. This method was compared for sensitivity of detection of escaped aerosols with a published method for aerosol detection which utilizes a UV-APS aerodynamic particle sizer and a UV-excitable dye. One of the advantages of the Cyclex-d system is the narrow-defined field of collection as compared to the standard Glo-Germ bead procedure, this means a smaller sampling area is used in the Cyclex-d impactor as compared to the AeroTech impactor. In addition, the sensitivity of detection was found to be better using the Cyclex-d collection device as compared to the standard Glo-Germ bead procedure. © 2018 International Society for Advancement of Cytometry.
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Aerosoles/análisis , Bioensayo/métodos , Citometría de Flujo/métodos , Sustancias Peligrosas/química , Separación Celular/métodos , Contención de Riesgos Biológicos/métodos , Contaminación de Equipos/prevención & control , Diseño de Equipo/métodos , Laboratorios , Microscopía Fluorescente/métodos , Microesferas , Tamaño de la PartículaRESUMEN
Leibovich et al. claim that number representations are non-existent early in life and that the associations between number and continuous magnitudes reside in stimulus confounds. We challenge both claims - positing, instead, that number is represented independently of continuous magnitudes already in infancy, but is nonetheless more deeply connected to other magnitudes through adulthood than acknowledged by the "sense of magnitude" theory.
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CogniciónRESUMEN
Flow cytometric cell sorting of biological specimens has become prevalent in basic and clinical research laboratories. These specimens may contain known or unknown infectious agents, necessitating precautions to protect instrument operators and the environment from biohazards arising from the use of sorters. To this end the International Society of Analytical Cytology (ISAC) was proactive in establishing biosafety guidelines in 1997 (Schmid et al., Cytometry 1997;28:99-117) and subsequently published revised biosafety standards for cell sorting of unfixed samples in 2007 (Schmid et al., Cytometry Part A J Int Soc Anal Cytol 2007;71A:414-437). Since their publication, these documents have become recognized worldwide as the standard of practice and safety precautions for laboratories performing cell sorting experiments. However, the field of cytometry has progressed since 2007, and the document requires an update. The new Standards provides guidance: (1) for laboratory design for cell sorter laboratories; (2) for the creation of laboratory or instrument specific Standard Operating Procedures (SOP); and (3) on procedures for the safe operation of cell sorters, including personal protective equipment (PPE) and validation of aerosol containment.
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Separación Celular/métodos , Citometría de Flujo , Administración de la Seguridad/normas , Sociedades Científicas , Separación Celular/normas , Contaminación de Equipos , Citometría de Flujo/métodos , Citometría de Flujo/normas , Sustancias Peligrosas , Humanos , Laboratorios/normas , Salud LaboralRESUMEN
Background: The Bicycle® toxin conjugate BT5528 is a novel peptide therapeutic conjugated to the cytotoxic agent monomethyl auristatin E (MMAE). A bioanalytical assay was developed to quantify BT5528 and unconjugated MMAE in human plasma. Methodology: BT5528 quantitation used a protein precipitation procedure followed by LC-MS/MS detection. Quantitation of MMAE required a selective offline and online solid-phase extraction with detection via LC-MS/MS. Results: BT5528 was quantified over the assay range of 5-2500 ng/ml and free MMAE was quantified over the assay range of 0.05-50 ng/ml. Conclusion: Bioanalytical methods were used in the bioanalysis of intact BT5528 and released MMAE, in a phase I/IIa clinical trial; to date, over 2000 human patient samples have been analyzed.
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Antineoplásicos , Inmunoconjugados , Inmunotoxinas , Oligopéptidos , Humanos , Cromatografía Liquida/métodos , Inmunotoxinas/análisis , Inmunoconjugados/análisis , Espectrometría de Masas en Tándem/métodos , CiclismoRESUMEN
Recent research investigating the languagethought interface in the spatial domain points to representations of the horizontal and vertical dimensions that closely resemble those posited by Jeffery et al. However, the findings suggest that such representations, rather than being tied to navigation, may instead reflect more general properties of the perception of space.
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Cognición/fisiología , Modelos Neurológicos , Percepción Espacial/fisiología , Conducta Espacial , Animales , HumanosRESUMEN
Although the representations underlying spatial language are often assumed to be schematic in nature, empirical evidence for a schematic format of representation is lacking. In this research, we investigate the psychological reality of such a format, using simulated motion during scene processing--previously linked to schematization--as a diagnostic. One group of participants wrote a verbal description of a scene and then completed a change detection task assessing simulated motion, while another group completed only the latter task. We expected that effects of simulated motion would be stronger following language use than not, and specifically following the use of spatial, relative to non-spatial, language. Both predictions were supported. Further, the effect of language was scene independent, suggesting that language may encourage a general mode of schematic construal. The study and its findings illustrate a novel approach to examining the perceptual properties of mental representations.
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Lenguaje , Prueba de Realidad , Percepción Espacial/fisiología , Conducta Espacial/fisiología , Adaptación Psicológica , Análisis de Varianza , Femenino , Humanos , Masculino , Movimiento (Física) , Estudiantes , UniversidadesRESUMEN
Detection of cell surface molecules labeled by monoclonal or polyclonal antibodies conjugated to a fluorochrome is the most widely used application of flow cytometry. Here, we present protocols for tagging monoclonal antibodies with fluorescein, biotin, Texas Red, and phycobiliproteins. In addition, we provide a procedure for preparing a PE-Texas Red tandem conjugate dye that can then be used for antibody conjugation. These protocols enable investigators to label antibodies of their choice with multiple fluorochromes and permit more combinations of antibodies for multicolor flow applications. © 2023 Wiley Periodicals LLC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA. Basic Protocol 1: Labeling an antibody with fluorescein isothiocyanate (FITC) Basic Protocol 2: Labeling an antibody with long-armed biotin Basic Protocol 3: Labeling an antibody with Texas Red-X Basic Protocol 4: Labeling an antibody with a synthetic organic fluor kit Basic Protocol 5: Labeling an antibody with phycobiliproteins Basic Protocol 6: Conjugation of Texas Red to R-phycoerythrin to produce an energy transfer fluorochrome.
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Anticuerpos Monoclonales , Colorantes Fluorescentes , Humanos , Biotina , Fluoresceína , FicoeritrinaRESUMEN
We join others in envisioning a future for affective science that addresses society's most pressing needs. To move toward this vision, we consider a research paradigm that emerged in other disciplines: use-inspired basic research. This paradigm transcends the traditional basic-applied dichotomy, which pits the basic goal of fundamental scientific understanding against the applied goal of use in solving social problems. In reality, these goals are complementary, and use-inspired basic research advances them simultaneously. Here, we build a case for use-inspired basic research-how it differs from traditional basic science and why affective scientists should engage in it. We first examine how use-inspired basic research challenges problematic assumptions of a strict basic-applied dichotomy. We then discuss how it is consistent with advances in affective science that recognize context specificity as the norm and consider ethical issues of use being a complementary goal. Following this theoretical discussion, we differentiate the implementation of use-inspired basic research from that of traditional basic science. We draw on examples from recent research to illustrate differences: social problems as a starting point, stakeholder and community engagement, and integration of research and service. In conclusion, we invite affective scientists to embrace the "lab meets world" perspective of use-inspired basic research as a promising pathway to real-world impact.
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Aim: A sensitive and selective method for the determination of PF-07059013 in dried blood collected by Mitra™ tips was developed and qualified from 50 to 50,000 ng/ml. Materials & methods: PF-07059013 is isolated from 10 µl of human dried blood by extraction with methanol and analyzed by HPLC-MS/MS. Results & conclusions: In addition to routine validation elements, impact of hematocrit and Mitra tip's lot-to-lot variation on assay accuracy were evaluated. The qualified method was used in one clinical study with excellent performance. Correlation coefficient between blood concentrations obtained from liquid-incurred blood samples and dried-incurred blood samples is 0.95. Clinical Trial Registration: NCT04323124 (ClinicalTrials.gov).
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Pruebas con Sangre Seca , Espectrometría de Masas en Tándem , Humanos , Espectrometría de Masas en Tándem/métodos , Pruebas con Sangre Seca/métodos , Manejo de Especímenes , Cromatografía Líquida de Alta Presión/métodos , HematócritoRESUMEN
Despite the recognition of potential aerosol hazards associated with cell sorting by the flow cytometry community, there has been no previous study that has thoroughly characterized the aerosols that can be produced by cell sorters. In this study, an aerodynamic particle sizer was used to determine the concentration and aerodynamic diameter (AD) of aerosols produced by a FACS Aria II cell sorter under various conditions. Aerosol containment and evacuation were also evaluated using this novel methodology. The results showed that high concentrations of aerosols in the range of 1-3 µm can be produced in fail mode and that with decreased sheath pressure, aerosol concentration decreased and AD increased. Although the engineering controls of the FACS Aria II for containment were effective, sort chamber evacuation of aerosols following a simulated nozzle obstruction was ineffective. However, simple modifications to the FACS Aria II are described that greatly improved sort chamber aerosol evacuation. The results of this study will facilitate the risk assessment of cell sorting potentially biohazardous samples by providing much needed data regarding aerosol production and containment.
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Aerosoles , Contención de Riesgos Biológicos , Contaminación de Equipos/prevención & control , Sustancias Peligrosas/análisis , Salud Laboral , Seguridad de Equipos , Citometría de Flujo/métodos , Humanos , Exposición Profesional , Traumatismos OcupacionalesRESUMEN
Converging behavioral and neural evidence suggests that numerical representations are mentally organized in left-to-right orientation. Here we show that this format of spatial organization extends to emotional expression. In Experiment 1, right-side responses became increasingly faster as number (represented by Arabic numerals) or happiness (depicted in facial stimuli) increased, for judgments completely unrelated to magnitude. Additional experiments suggest that magnitude (i.e., more/less relations), not valence (i.e., positive/negative), underlies left-to-right orientation of emotional expression (Experiment 2), and that this orientation accommodates to the context-relevant emotion (e.g., happier faces are more rightward when judged on happiness, but more leftward when judged on angriness; Experiment 3). These findings show that people automatically extract magnitude from a variety of stimuli, representing such information in common left-to-right format, perhaps reflecting a mental magnitude line. We suggest that number is but one dimension in a hyper-general representational system uniting disparate dimensions of magnitude and likely subserved by common neural mechanisms in posterior parietal cortex.
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Encéfalo/fisiología , Emociones/fisiología , Imaginación/fisiología , Orientación/fisiología , Percepción Espacial/fisiología , Adulto , Femenino , Humanos , Masculino , Tiempo de Reacción/fisiologíaRESUMEN
Accumulating evidence suggests that different magnitudes (e.g., number, size, and duration) are spatialized in the mind according to a common left-right metric, consistent with a generalized system for representing magnitude. A previous study conducted by two of us (Holmes & Lourenco, ) provided evidence that this metric extends to the processing of emotional magnitude, or the intensity of emotion expressed in faces. Recently, however, Pitt and Casasanto () showed that the earlier effects may have been driven by a left-right mapping of mouth size rather than emotional magnitude, and they found no evidence for an emotional magnitude mapping when using words as stimuli. Here, we report two new experiments that further examine these conclusions. In Experiment 1, using face stimuli with mouths occluded, we replicate the original finding: Less emotional faces were associated with the left and more emotional faces with the right. However, we also find that people can reliably infer the sizes of the occluded mouths, and that these inferred mouth sizes can explain the observed left-right mapping. In Experiment 2, we show that comparative judgments of emotional words yield a left-right mapping of emotional magnitude not attributable to stimulus confounds. Based on these findings, we concur with Pitt and Casasanto that faces pose challenges for isolating the forces driving spatialization, but we suggest that emotional magnitude, when assessed using unconfounded stimuli in a sufficiently sensitive task, may indeed be spatialized as originally proposed. Suggestions for further research on the spatialization of emotional magnitude are discussed.
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Emociones/fisiología , Expresión Facial , Juicio/fisiología , Percepción Social , Femenino , Humanos , Masculino , Estimulación Luminosa , Tiempo de Reacción/fisiologíaRESUMEN
Signal transducer and activator of transcription (STAT1)1 gain of function (GOF) pathogenic variants have been associated with increased levels of phosphorylated STAT1 and STAT1-dependent cellular responses. Delayed dephosphorylation was proposed as the underlying mechanism leading to the characteristically raised pSTAT1 levels. We examined the levels of STAT1 protein and message as well as rates of STAT1 phosphorylation, dephosphorylation, and degradation associated with STAT1 GOF pathogenic variants. Fresh peripheral blood mononuclear cells (PBMC) from 14 STAT1 GOF patients carrying 10 different pathogenic variants in the coiled-coil, DNA binding, and SH2 domains and healthy donors were used to study STAT1 levels and phosphorylation (pSTAT1) following IFNγ and IFNα stimulation. STAT1 protein levels were measured by flow cytometry and immunoblot. STAT1 mRNA levels were measured using quantitative reverse transcription PCR. STAT1 protein degradation was studied using cycloheximide. Patient IFNγ and IFNα induced peak pSTAT1 was higher than in healthy controls. The velocity of pSTAT1 dephosphorylation after treatment of IFNγ stimulated CD14+ monocytes with the Janus Kinase (JAK)-inhibitor ruxolitinib was significantly faster in patient cells. STAT1 protein levels in patient CD14+ monocytes and CD3+ T cells were higher than in healthy donors. There was a strong and positive correlation between CD14+ STAT1 protein levels and peak pSTAT1 levels. Patient fresh PBMC STAT1 mRNA levels were increased at rest and after 16 h of incubation. STAT1 protein degradation was similar in patient and healthy volunteer cells. Patient IFNγ receptors 1 and 2 and JAK2 levels were normal. One patient in our cohort was treated with the oral JAK inhibitor ruxolitinib. Treatment was associated with normalization of both STAT1 protein and peak pSTAT1 levels. After JAK inhibitor treatment was stopped the patient's CD14+ monocyte STAT1 protein and peak phosphorylation levels increased proportionally. These findings suggest that patients with STAT1 GOF mutations have higher levels of total STAT1 protein, leading to high levels of pSTAT1 after stimulation, despite rapid STAT1 dephosphorylation and normal degradation.
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Enfermedades Autoinmunes/metabolismo , Mutación con Ganancia de Función/genética , Leucocitos Mononucleares/inmunología , Micosis/metabolismo , Factor de Transcripción STAT1/genética , Adolescente , Adulto , Enfermedades Autoinmunes/genética , Células Cultivadas , Niño , Preescolar , Femenino , Humanos , Masculino , Persona de Mediana Edad , Micosis/genética , Fosforilación , Proteolisis , Regulación hacia Arriba , Adulto JovenRESUMEN
Parts of the body are often embedded in the structure of compound words, such as heartbreak and brainchild. We explored the relationships between the semantics of compounds and their constituent body parts, asking whether these relationships are largely arbitrary or instead reflect deeper metaphorical mappings shared across languages and cultures. In three studies, we found that U.S. English speakers associated the English translation equivalents of Chinese compounds with their constituent body parts at rates well above chance, even for compounds with highly abstract meanings and even when accounting for the semantic relatedness of the compounds and body parts. English speakers in India and Chinese speakers in Hong Kong showed similar intuitions about these associations. Our results suggest that the structure of compound words can provide insight into cross-culturally shared ways of connecting meaning to the body.
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Cuerpo Humano , Lenguaje , Metáfora , Adulto , Comparación Transcultural , Femenino , Hong Kong , Humanos , India , Masculino , Estados Unidos , Adulto JovenRESUMEN
BACKGROUND: Vascular endothelial growth factor has been found to accelerate compensatory lung growth after left pneumonectomy in mice. The aim of this study was to determine the natural history and the effects of vascular endothelial growth factor on compensatory lung growth in a large animal model. METHODS: To determine the natural history of compensatory lung growth, female Yorkshire piglets underwent a left pneumonectomy on days of life 10-11. Tissue harvest and volume measurement of the right lung were performed at baseline (nâ¯=â¯5) and on postoperative days 7 (nâ¯=â¯5), 14 (nâ¯=â¯4), and 21 (nâ¯=â¯5). For pharmacokinetic studies, vascular endothelial growth factor was infused via a central venous catheter, with plasma vascular endothelial growth factor levels measured at various time points. To test the effect of vascular endothelial growth factor on compensatory lung growth, 26 female Yorkshire piglets underwent a left pneumonectomy followed by daily infusion of vascular endothelial growth factor at 200 µg/kg or isovolumetric 0.9% NaCl (saline control). Lungs were harvested on postoperative day 7 for volume measurement and morphometric analyses. RESULTS: Compared with baseline, right lung volume after left pneumonectomy increased by factors of 2.1 ± 0.6, 3.3 ± 0.6, and 3.6 ± 0.4 on postoperative days 7, 14, and 21, respectively. The half-life of VEGF ranged from 89 to 144 minutes. Lesser doses of vascular endothelial growth factor resulted in better tolerance, volume of distribution, and clearance. Compared with the control group, piglets treated with vascular endothelial growth factor had greater lung volume (P < 0.0001), alveolar volume (Pâ¯=â¯0.001), septal surface area (Pâ¯=â¯0.007) and total alveolar count (Pâ¯=â¯0.01). CONCLUSION: Vascular endothelial growth factor enhanced alveolar growth in neonatal piglets after unilateral pneumonectomy.
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Pulmón/crecimiento & desarrollo , Factor A de Crecimiento Endotelial Vascular/farmacocinética , Animales , Animales Recién Nacidos , Biometría , Evaluación Preclínica de Medicamentos , Femenino , Pulmón/efectos de los fármacos , Neumonectomía , Proteínas Recombinantes , Porcinos , Factor A de Crecimiento Endotelial Vascular/administración & dosificaciónRESUMEN
Categories can affect our perception of the world, rendering between-category differences more salient than within-category ones. Across many studies, such categorical perception (CP) has been observed for the basic-level categories of one's native language. Other research points to categorical distinctions beyond the basic level, but it does not demonstrate CP for such distinctions. Here we provide such a demonstration. Specifically, we show CP in English speakers for the non-basic distinction between "warm" and "cool" colors, claimed to represent the earliest stage of color lexicon evolution. Notably, the advantage for discriminating colors that straddle the warm-cool boundary was restricted to the right visual field-the same behavioral signature previously observed for basic-level categories. This pattern held in a replication experiment with increased power. Our findings show that categorical distinctions beyond the basic-level repertoire of one's native language are psychologically salient and may be spontaneously accessed during normal perceptual processing.