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1.
Sci Rep ; 10(1): 934, 2020 01 22.
Artículo en Inglés | MEDLINE | ID: mdl-31969626

RESUMEN

Josephson junctions act as a natural spiking neuron-like device for neuromorphic computing. By leveraging the advances recently demonstrated in digital single flux quantum (SFQ) circuits and using recently demonstrated magnetic Josephson junction (MJJ) synaptic circuits, there is potential to make rapid progress in SFQ-based neuromorphic computing. Here we demonstrate the basic functionality of a synaptic circuit design that takes advantage of the adjustable critical current demonstrated in MJJs and implement a synaptic weighting element. The devices were fabricated with a restively shunted Nb/AlOx-Al/Nb process that did not include MJJs. Instead, the MJJ functionality was tested by making multiple circuits and varying the critical current, but not the external shunt resistance, of the oxide Josephson junction that represents the MJJ. Experimental measurements and simulations of the fabricated circuits are in good agreement.

2.
Appl Radiat Isot ; 72: 128-32, 2013 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-23208243

RESUMEN

A simple synthesis of the dopamine transporter ligand [(18)F]FECNT with high radiochemical yield and short synthesis time, suitable for routine production is reported. Reaction of 2ß-carbomethoxy-3ß-(4-chlorophenyl)nortropane with [(18)F]2-fluoroethyl triflate ([(18)F]FEtOTf) at room temperature for 4 min provided [(18)F]FECNT in 84% decay corrected radiochemical yield. Since [(18)F]FEtOTf was prepared from [(18)F]2-fluoroethyl bromide that was isolated from its starting material, formation of unwanted side products and the amount of expensive precursor used could be greatly reduced. The overall radiochemical yields of [(18)F]FECNT were 40% (n=29) and the total synthesis time was ca. 100 min. The average specific activity of [(18)F]FECNT was 377.4 GBq/µmol (10.2 Ci/µmol).


Asunto(s)
Proteínas de Transporte de Dopamina a través de la Membrana Plasmática/análisis , Radioisótopos de Flúor/química , Nortropanos/síntesis química , Cromatografía Líquida de Alta Presión
3.
Synapse ; 65(12): 1309-18, 2011 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-21674627

RESUMEN

OBJECTIVE: [F-18]Nifene is a PET radioligand developed to image α4ß2* nicotinic acetylcholine receptors (nAChR) in the brain. This work assesses the in vivo binding and imaging characteristics of [F-18]nifene in rhesus monkeys for the development of PET experiments examining nAChR binding. METHODS: Dynamic PET imaging experiments with [F-18]nifene were acquired in four anesthetized Macaca mulatta (rhesus) monkeys using a microPET P4 scanner. Data acquisition was initiated with a bolus injection of 109 ± 17 MBq [F-18]nifene and the time course of the radioligand in the brain was measured for up to 120 min. For two experiments, a displacement dose of (-)nicotine (0.03 mg kg(-1) , i.v.) was given 45-60 min post injection and followed 30 min later with a second [F-18]nifene injection to measure radioligand nondisplaceable uptake. Time activity curves were extracted in the regions of the antereoventral thalamus (AVT), lateral geniculate nucleus region (LGN), frontal cortex, and the cerebellum (CB). RESULTS: The highest levels of [F-18]nifene uptake were observed in the AVT and LGN. Target-to-CB ratios reached maximum values of 3.3 ± 0.4 in the AVT and 3.2 ± 0.3 in the LGN 30-45 min postinjection. Significant binding of [F-18]nifene was observed in the subiculum, insula cortex, temporal cortex, cingulate gyrus, frontal cortex, striatum, and midbrain areas. The (-)nicotine displaced bound [F-18]nifene to near background levels within 15 min postdrug injection. No discernable displacement was observed in the CB, suggesting its potential as a reference region. Logan graphical estimates using the CB as a reference region yielded binding potentials of 1.6 ± 0.2 in the AVT and 1.3 ± 0.1 in the LGN. The postnicotine injection displayed uniform nondisplaceable uptake of [F-18]nifene throughout gray and white brain matter. CONCLUSIONS: [F-18]Nifene exhibits rapid equilibration and a moderately high target to background binding profile in the α4ß2* nAChR rich regions of the brain, thus providing favorable imaging characteristics as a PET radiotracer for nAChR assay.


Asunto(s)
Piridinas/metabolismo , Pirroles/metabolismo , Receptores Nicotínicos/metabolismo , Animales , Sitios de Unión/fisiología , Encéfalo/diagnóstico por imagen , Encéfalo/metabolismo , Femenino , Macaca mulatta , Masculino , Neuroimagen/métodos , Tomografía de Emisión de Positrones/métodos , Unión Proteica/fisiología
4.
Synapse ; 65(7): 592-600, 2011 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-21484878

RESUMEN

UNLABELLED: [F-18]Mefway was developed to provide an F-18 labeled positron emission tomography (PET) neuroligand with high affinity for the serotonin 5-HT(1A) receptor to improve the in vivo assessment of the 5-HT(1A) system. The goal of this work was to compare the in vivo kinetics of [F-18]mefway, [F-18]MPPF, and [C-11]WAY100635 in the rhesus monkey. METHODS: Each of four monkeys were given bolus injections of [F-18]mefway, [C-11]WAY100635, and [F-18]MPPF and scans were acquired with a microPET P4 scanner. Arterial blood was sampled to assay parent compound throughout the time course of the PET experiment. Time activity curves were extracted in the high 5-HT(1A) binding areas of the anterior cingulate cortex (ACG), mesial temporal cortex, raphe nuclei, and insula cortex. Time activity curves were also extracted in the cerebellum, which was used as a reference region. The in vivo kinetics of the radiotracers were compared based on the nondisplaceable distribution volume (V(ND) ) and binding potential (BP(ND) ). RESULTS: At 30 min, the fraction of radioactivity in the plasma due to parent compound was 19%, 28%, and 29% and cleared from the arterial plasma at rates of 0.0031, 0.0078, and 0.0069 (min⁻¹) ([F-18]mefway, [F-18]MPPF, [C-11]WAY100635). The BP(ND) in the brain regions were mesial temporal cortex: 7.4 ± 0.6, 3.1 ± 0.4, 7.0 ± 1.2, ACG: 7.2 ± 1.2, 2.1 ± 0.2, 7.9 ± 1.2; raphe nuclei: 3.7 ± 0.6, 1.3 ± 0.3, 3.3 ± 0.7; and insula cortex: 4.2 ± 0.6, 1.2 ± 0.1, 4.7 ± 1.0 for [F-18]mefway, [F-18]MPPF, and [C-11]WAY100635 respectively. CONCLUSIONS: In the rhesus monkey, [F-18]mefway has similar in vivo kinetics to [C-11]WAY100635 and yields greater than 2-fold higher BP(ND) than [F-18]MPPF. These properties make [F-18]mefway a promising radiotracer for 5-HT(1A) assay, providing higher counting statistics and a greater dynamic range in BP(ND).


Asunto(s)
Encéfalo/diagnóstico por imagen , Piperazinas/farmacocinética , Piridinas/farmacocinética , Radiofármacos/farmacocinética , Animales , Radioisótopos de Carbono/farmacocinética , Radioisótopos de Flúor/farmacocinética , Macaca mulatta , Tomografía de Emisión de Positrones/métodos
5.
Neurotoxicol Teratol ; 26(2): 169-78, 2004.
Artículo en Inglés | MEDLINE | ID: mdl-15019951

RESUMEN

This study examined the striatal dopamine system integrity and associated behavior in 5- to 7-year-old rhesus monkeys born from mothers that experienced stress and/or consumed moderate levels of alcohol during pregnancy. Thirty-one young adult rhesus monkeys were derived from females randomly assigned to one of four groups: (1) control group that consumed isocaloric sucrose solution throughout gestation; (2) stress group that experienced prenatal stress (10-min removal from home cage and exposure to three random loud noise bursts, gestational days 90 through 145); (3) alcohol group that consumed alcohol (0.6 g/kg/day) throughout gestation; or (4) combined alcohol plus stress group that received both treatments. The subjects were assessed for striatal dopamine system function using positron emission tomography (PET), in which the dopamine (DA)-rich striatum was evaluated in separate scans for the trapping of [(18)F]-Fallypride (FAL) and 6-[(18)F]fluoro-m-tyrosine (FMT) to assess dopamine D2 receptor binding potential (BP) and DA synthesis via dopa decarboxylase activity, respectively. Subjects were previously assessed for non-matching-to-sample (NMS) task acquisition, with ratings of behavioral inhibition, stereotypies, and activity made after each NMS testing session. Subjects from prenatal stress conditions (Groups 2 and 4) showed an increase in the ratio of striatal dopamine D2 receptor BP and DA synthesis compared to controls (Group 1). An increase in the radiotracer distribution volume ratios (DVRs), which is used to evaluate the balance between striatal DA synthesis and receptor availability, respectively, was significantly correlated with less behavioral inhibition. The latter supports a hypothesis linking striatal function to behavioral inhibitory control.


Asunto(s)
Conducta Animal/efectos de los fármacos , Depresores del Sistema Nervioso Central/farmacología , Dopamina/metabolismo , Etanol/farmacología , Feto/efectos de los fármacos , Efectos Tardíos de la Exposición Prenatal , Estrés Psicológico , Análisis de Varianza , Animales , Animales Recién Nacidos , Cognición/fisiología , Femenino , Inhibición Psicológica , Macaca mulatta , Masculino , Actividad Motora/efectos de los fármacos , Embarazo , Distribución Aleatoria , Conducta Estereotipada/efectos de los fármacos , Tomografía Computarizada de Emisión/métodos
7.
Oncogene ; 20(51): 7524-35, 2001 Nov 08.
Artículo en Inglés | MEDLINE | ID: mdl-11709724

RESUMEN

Using the established quail cell line Q/d3 conditionally transformed by the v-jun oncogene, cDNA clones (TOJ2, TOJ3, TOJ5, TOJ6) were isolated by representational difference analysis (RDA) that correspond to genes which were induced immediately upon conditional activation of v-jun. One of these genes, TOJ3, is immediately and specifically activated after doxycycline-mediated v-jun induction, with kinetics similar to the induction of well characterized direct AP-1 target genes. TOJ3 is neither activated upon conditional activation of v-myc, nor in cells or cell lines non-conditionally transformed by oncogenes other than v-jun. Sequence analysis revealed that the TOJ3-specific cDNA encodes a 530-amino acid protein with significant sequence similarities to the murine or human microspherule protein 1 (MCRS1, MSP58), a nucleolar protein that directly interacts with the ICP22 regulatory protein from herpes simplex virus 1 or with p120, a proliferation-related protein expressed at high levels in most human malignant tumor cells. Similar to its mammalian counterparts, the TOJ3 protein contains a bipartite nuclear localization motif and a forkhead associated domain (FHA). Using polyclonal antibodies directed against a recombinant amino-terminal TOJ3 protein segment, the activation of TOJ3 in jun-transformed fibroblasts was also demonstrated at the protein level by specific detection of a polypeptide with an apparent M(r) of 65 000. Retroviral expression of the TOJ3 gene in quail or chicken embryo fibroblasts induces anchorage-independent growth, indicating that the immediate activation of TOJ3 in fibroblasts transformed by the v-jun oncogene contributes to cell transformation.


Asunto(s)
Proteínas Aviares , Proteínas Portadoras/metabolismo , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/metabolismo , Proteínas Nucleares/metabolismo , Proteína Oncogénica p65(gag-jun)/metabolismo , Secuencia de Aminoácidos , Animales , Anticuerpos/metabolismo , Secuencia de Bases , Northern Blotting , Proteínas Portadoras/química , Nucléolo Celular/metabolismo , Transformación Celular Neoplásica , Embrión de Pollo , Cromatografía , Clonación Molecular , Coturnix , ADN/metabolismo , ADN Complementario/metabolismo , Doxiciclina/farmacología , Activación Enzimática , Fibroblastos/metabolismo , Humanos , Cinética , Ratones , Modelos Genéticos , Datos de Secuencia Molecular , Proteínas Nucleares/química , Pruebas de Precipitina , Unión Proteica , Biosíntesis de Proteínas , Estructura Terciaria de Proteína , Proteínas/metabolismo , ARN/metabolismo , Proteínas Recombinantes/metabolismo , Retroviridae/genética , Análisis de Secuencia de ADN , Homología de Secuencia de Aminoácido , Factores de Tiempo , Transcripción Genética , Activación Transcripcional , Células Tumorales Cultivadas
8.
Alcohol Clin Exp Res ; 25(9): 1383-92, 2001 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-11584160

RESUMEN

BACKGROUND: Although high-dose prenatal alcohol exposure is related to cognitive and behavioral impairments in children and adolescents with fetal alcohol syndrome, there is relatively little research on the effects of moderate drinking during pregnancy. We examined learning, memory, and behavior in adolescent rhesus monkeys prenatally exposed to moderate levels of alcohol, psychological stress, or both alcohol and stress. METHODS: Forty adolescent rhesus monkey subjects were derived from four groups of female rhesus monkeys that (1) consumed alcohol throughout gestation; (2) experienced prenatal stress; (3) experienced prenatal stress and alcohol consumption; or (4) control group (no alcohol, no stress). The subjects were assessed for number of trials required to reach 90% criterion of correct responses on nonmatching-to-sample task (NMS), followed by trials with delays of 30, 60, or 120 sec. Ratings of behavior during testing were made after each session. RESULTS: Subjects exposed to moderate prenatal alcohol required significantly more trials to reach criterion on the acquisition phase of the NMS task but had no difficulty with delays. Prenatally stressed monkeys showed lower response inhibition or less behavioral restraint, whereas prenatal alcohol plus stress monkeys showed higher activity level and stereotypies compared with controls. High scores on neonatal measures of orientation (attending to novel stimuli) and motor maturity and low scores on irritability, activity, stereotypies, and impulsivity during acquisition were correlated with fewer trials to criterion on acquisition of NMS. CONCLUSIONS: NMS trials required to reach criterion and behavior during testing are sensitive to moderate-level prenatal alcohol exposure in monkeys. The most adverse behavioral outcomes (hyperactivity and stereotypies) were associated with prenatal alcohol plus stress, raising concerns that environmental stress might provide the context within which fetal alcohol exposure could promote adverse behavioral outcomes. These effects occurred in the absence of either facial deformities or retarded physical growth.


Asunto(s)
Conducta Animal/efectos de los fármacos , Etanol/farmacología , Aprendizaje/efectos de los fármacos , Efectos Tardíos de la Exposición Prenatal , Animales , Etanol/administración & dosificación , Femenino , Conducta Impulsiva , Genio Irritable , Macaca mulatta , Actividad Motora , Embarazo , Conducta Estereotipada , Estrés Fisiológico/complicaciones
9.
Alcohol Clin Exp Res ; 25(8): 1238-45, 2001 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-11505056

RESUMEN

BACKGROUND: Moderate prenatal alcohol exposure can contribute to neurodevelopmental deficits in nonhuman primate offspring. The purpose of this study was to examine the effects of gestational timing of alcohol exposure on neurobehavior with a nonhuman primate model. METHODS: Sixty-three rhesus monkey infants (Macaca mulatta), from four groups of females, were assessed: (1) an early alcohol-exposed group, in which mothers voluntarily consumed alcohol on gestational days 0 through 50; (2) a mid to late gestation alcohol-exposed group, in which mothers consumed an identical dose on gestation days 50 through 135; (3) a continuous-exposure group, in which mothers consumed an identical dose on days 0 through 135 or days 0 through 165; and (4) controls, in which mothers voluntarily consumed an isocaloric control solution on gestational days 0 through 50, 50 through 135, 0 through 135, or 0 through 165. Data were obtained on offspring for measures of growth and neurobehavior. RESULTS: There were no effects of alcohol on birthweight, gestation length, or ponderal index. Prenatal exposure to alcohol during early gestation significantly decreased scores on infant neurobehavioral tests overall in multivariate tests, after controlling for birthweight. Univariate tests showed that early gestation alcohol exposure was related to reductions in infant orientation and motor maturity. Mid- to late-gestation exposure also resulted in a reduction in motor maturity but did not affect overall neurobehavioral performance in the multivariate tests. CONCLUSIONS: Early-gestation alcohol exposure is as deleterious to neonatal neurobehavior as late-gestation or continuous exposure. Moreover, neurobehavior seems to be a more sensitive marker of early-gestation moderate alcohol exposure than growth parameters. Women who are attempting to become pregnant should minimize frequent social drinking, because subtle neurodevelopmental effects to the fetus may be induced before pregnancy is detected.


Asunto(s)
Animales Recién Nacidos , Etanol/administración & dosificación , Intercambio Materno-Fetal , Resultado del Embarazo , Animales , Conducta Animal , Peso al Nacer , Femenino , Edad Gestacional , Macaca mulatta , Masculino , Conducta Materna , Actividad Motora , Embarazo , Factores de Tiempo
10.
J Mol Biol ; 307(5): 1395-410, 2001 Apr 13.
Artículo en Inglés | MEDLINE | ID: mdl-11292350

RESUMEN

The protein product (c-Myc) of the protooncogene c-myc is a transcriptional regulator playing a key role in cellular growth, differentiation, and apoptosis. Deregulated myc genes, like the transduced retroviral v-myc allele, are oncogenic and cause cell transformation. The C-terminal bHLHZip domain of v-Myc, encompassing protein dimerization (helix-loop-helix, leucine zipper) and DNA contact (basic region) surfaces, was expressed in bacteria as a highly soluble p15(v-myc )recombinant protein. Dissociation constants (K(d)) for the heterodimer formed with the recombinant bHLHZip domain of the Myc binding partner Max (p14(max)) and for the Myc-Max-DNA complex were estimated using circular dichroism (CD) spectroscopy and quantitative electrophoretic mobility shift assay (EMSA). Multi-dimensional NMR spectroscopy was used to characterize the solution structural and dynamic properties of the v-Myc bHLHZip domain. Significant secondary chemical shifts indicate the presence of two separated alpha-helical regions. The C-terminal leucine zipper region forms a compact alpha-helix, while the N-terminal basic region exhibits conformational averaging with substantial alpha-helical content. Both helices lack stabilizing tertiary side-chain interactions and represent exceptional examples for loosely coupled secondary structural segments in a native protein. These results and CD thermal denaturation data indicate a monomeric state of the v-Myc bHLHZip domain. The (15)N relaxation data revealed backbone mobilities which corroborate the existence of a partially folded state, and suggest a "beads-on-a-string" motional behaviour of the v-Myc bHLHZip domain in solution. The preformation of alpha-helical regions was confirmed by CD thermal denaturation studies, and quantification of the entropy changes caused by the hydrophobic effect and the reduction of conformational entropy upon protein dimerization. The restricted conformational space of the v-Myc bHLHZip domain reduces the entropy penalty associated with heterodimerization and allows rapid and accurate recognition by the authentic Myc binding partner Max.


Asunto(s)
ADN/metabolismo , Proteína Oncogénica p55(v-myc)/química , Proteína Oncogénica p55(v-myc)/metabolismo , Factores de Transcripción/química , Factores de Transcripción/metabolismo , Secuencia de Aminoácidos , Animales , Factores de Transcripción con Cremalleras de Leucina de Carácter Básico , Pollos , Dicroismo Circular , ADN/genética , Proteínas de Unión al ADN/química , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Dimerización , Entropía , Secuencias Hélice-Asa-Hélice , Leucina Zippers , Espectroscopía de Resonancia Magnética , Datos de Secuencia Molecular , Movimiento (Física) , Proteína Oncogénica p55(v-myc)/genética , Desnaturalización Proteica , Pliegue de Proteína , Estructura Secundaria de Proteína , Estructura Terciaria de Proteína , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Alineación de Secuencia , Solubilidad , Relación Estructura-Actividad , Temperatura , Factores de Transcripción/genética
11.
Stress ; 4(3): 183-93, 2001 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-22432139

RESUMEN

In this paper we review three prospective longitudinal studies from our laboratory examining the effects of prenatal stress on early neuro behavior, stress reactivity and learning performance in rhesus monkeys. Either a noise stressor or ACTH treatment was administered to pregnant monkeys during specific periods of pregnancy and offspring were examined repeatedly across development. In all three studies, the prenatally stressed monkeys showed reduced attention and impaired neuromotor functioning during the first month of life compared to controls from undisturbed pregnancies. When the monkeys were separated from their mothers or peers at 6-8 months of age, prenatally stressed monkeys exhibited more disturbance behavior and showed hypothalamic-pituitary-adrenal axis dysregulation. During adolescence, they exhibited impairments in learning, compared to controls.


Asunto(s)
Conducta Animal , Aprendizaje , Conducta Materna , Complicaciones del Embarazo/etiología , Efectos Tardíos de la Exposición Prenatal , Estrés Psicológico/complicaciones , Factores de Edad , Animales , Ansiedad de Separación/metabolismo , Ansiedad de Separación/psicología , Atención , Modelos Animales de Enfermedad , Femenino , Edad Gestacional , Humanos , Sistema Hipotálamo-Hipofisario/metabolismo , Macaca mulatta , Actividad Motora , Sistema Hipófiso-Suprarrenal/metabolismo , Embarazo , Complicaciones del Embarazo/psicología , Estrés Psicológico/psicología
12.
Mol Cell Neurosci ; 18(5): 458-72, 2001 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-11922138

RESUMEN

Neuronal differentiation is regulated by both positive and negative regulatory factors; however, precisely how these factors interact to regulate retinogenesis is still unclear. We have examined the ability of the Notch pathway to modulate the function of the basic helix-loop-helix factor Xath5. Overexpression of Xath5 by RNA injection into cleavage-stage blastomeres promotes ectopic neurogenesis at neural plate stages and ganglion cell differentiation in the developing retina. We found that these activities of Xath5 could be inhibited by coexpression of activated Notch. Notch inhibition of Xath5 function was reversed by coexpression with the zinc finger protein X-MyT1. The Notch effector enhancer-of-split related 1 (ESR1) also blocked Xath5 activity but efficient inhibition by ESR1 required the DNA binding basic domain and the conserved WRPW motif. In addition, ESR1 inhibited the ability of Xath5 to directly activate the expression of XBrn3d, a transcription factor involved in retinal ganglion cell development. Xath5 could upregulate expression of X-Delta-1, ESR1, and ESR3, suggesting that Xath5 participates in a regulatory loop with the Notch pathway.


Asunto(s)
Diferenciación Celular/fisiología , Sistema Nervioso Central/embriología , Proteínas del Ojo/metabolismo , Proteínas de la Membrana/metabolismo , Neuronas/metabolismo , Retina/embriología , Proteínas de Saccharomyces cerevisiae , Factores de Transcripción/metabolismo , Proteínas de Xenopus , Xenopus laevis/embriología , Secuencias de Aminoácidos/genética , Animales , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico , Sitios de Unión/genética , Blastómeros/citología , Blastómeros/efectos de los fármacos , Blastómeros/metabolismo , Diferenciación Celular/efectos de los fármacos , Sistema Nervioso Central/citología , Sistema Nervioso Central/metabolismo , Proteínas de Unión al ADN/genética , Proteínas del Ojo/genética , Femenino , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Regulación del Desarrollo de la Expresión Génica/fisiología , Proteínas Fluorescentes Verdes , Péptidos y Proteínas de Señalización Intracelular , Proteínas Luminiscentes/genética , Proteínas de la Membrana/genética , Neuronas/citología , Proteínas Serina-Treonina Quinasas/genética , Proteínas Serina-Treonina Quinasas/metabolismo , Proteínas Tirosina Quinasas/genética , Proteínas Tirosina Quinasas/metabolismo , ARN Mensajero/genética , ARN Mensajero/farmacología , Receptores Notch , Retina/citología , Retina/metabolismo , Células Ganglionares de la Retina/citología , Células Ganglionares de la Retina/metabolismo , Factores de Transcripción/genética , Tubulina (Proteína)/metabolismo , Xenopus laevis/metabolismo
13.
Med Res Rev ; 19(4): 295-305, 1999 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-10398926

RESUMEN

A structural basis for activation and substrate specificity of src tyrosine kinases, and regulation of protein-protein association by tyrosine phosphorylation is described. Lyn, a src-family tyrosine kinase, recognizes and phosphorylates the immunoreceptor tyrosine-based activation motif, ITAM, a critical component in transmembrane signal transduction in hemopoietic cells. The structure of an ITAM peptide substrate bound to an active form of Lyn tyrosine kinase was determined by high-resolution NMR, and a model of the complex was generated using the crystallographic structure of Lck, a closely related Src-family kinase. The results provide a rationale for the conserved ITAM residues and specificity of Lyn, and suggest that substrate plays a role in stabilizing the kinase conformation optimal for catalysis. It is our hope that the Lck-ITAM peptide model complex will be useful in aiding structure-based drug design efforts that target substrate binding determinants in the design. Concerning the regulation of protein-protein association, we report on a complex between erythrocyte band 3 and two glycolytic enzymes, aldolase and glyceraldehyde-3-phosphate dehydrogenase. The formation of this complex is negatively regulated by tyrosine phosphorylation of band 3 by p72syk tyrosine kinase. In red blood cells, this association results in a decrease in glycolysis due to competitive inhibition of the glycolytic enzymes. The structure of band 3 recognized by the glycolytic enzymes was determined by solution NMR, and found to be a loop structure with tyrosine centrally positioned and excluded from intermolecular contact. This phosphorylation sensitive interaction, or PSI, loop may be the basis of a general mechanism for negative regulation through tyrosine phosphorylation.


Asunto(s)
Fosfotirosina/química , Fosfotirosina/fisiología , Señales de Clasificación de Proteína/química , Señales de Clasificación de Proteína/fisiología , Diseño de Fármacos , Humanos , Modelos Estructurales , Conformación Proteica
14.
Child Dev ; 70(2): 263-74, 1999.
Artículo en Inglés | MEDLINE | ID: mdl-10218255

RESUMEN

Previous studies have found that stressful events during pregnancy can influence the developing fetus, resulting in attentional and neuromotor problems. This prospective study examined whether periods of vulnerability exist for neurobehavioral impairments associated with prenatal stress, using a nonhuman primate model. Twenty-eight rhesus monkey infants were born to mothers in 3 groups: (1) early gestation stress involving mild psychological stress from gestational days 45-90, (2) mid-late gestation stress from days 90-145, and (3) undisturbed controls. Infants were separated from their mothers on days 4, 9, 15, and 22 (+/- 1) postpartum for growth and neurobehavioral assessments. Results indicated that infants from the early gestation stress condition weighed less than infants from mothers stressed during mid-late gestation. Moreover, whereas both groups scored lower than controls on measures of attention and neuromotor maturity, early gestation stress was associated with more pronounced and more pervasive motor impairments than mid-late gestation stress. These results suggest sensitivity to prenatal stress effects peaks during early gestation, tapering off during mid-late gestation. Clarifying the period of greatest vulnerability to prenatal stress moves toward elucidating the underlying mechanism for prenatal stress effects and may lead to more successful intervention and/or prevention.


Asunto(s)
Animales Recién Nacidos/crecimiento & desarrollo , Animales Lactantes/crecimiento & desarrollo , Discapacidades del Desarrollo/etiología , Exposición Materna/efectos adversos , Efectos Tardíos de la Exposición Prenatal , Estrés Psicológico/fisiopatología , Animales , Animales Recién Nacidos/psicología , Animales Lactantes/psicología , Peso al Nacer , Femenino , Manejo Psicológico , Hidrocortisona/sangre , Macaca mulatta , Masculino , Destreza Motora/fisiología , Orientación/fisiología , Embarazo , Estrés Psicológico/sangre
15.
Dev Psychopathol ; 10(3): 427-40, 1998.
Artículo en Inglés | MEDLINE | ID: mdl-9741675

RESUMEN

In this study, we assessed behavioral responses to social separation at 8 months of age and cerebrospinal fluid (CSF) concentrations of biogenic amines and metabolites at 8 and 18 months of age in 12 rhesus monkeys derived from either stressed or undisturbed pregnancies. Compared to controls from undisturbed pregnancies, prenatal stress-derived monkeys had higher concentrations of 3-methoxy-4-hydroxyphenylglycol (MHPG), and 3,4-dihydroxyphenylacetic acid in CSF than controls. Norepinephrine and MHPG response to stress were both correlated between 8 and 18 months of age. There were few group differences in behavior during social separation; however, several behavioral differences between groups were found when monkeys were reunited with cage mates. Prenatally stressed monkeys spent more time clinging to their surrogates and exploring (including eating and drinking), while controls showed more locomotion and social play with their cage mates. Collectively, our findings suggest that chronic unpredictable psychological stress during pregnancy has long-lasting effects on noradrenergic and dopaminergic activity and behavior in the offspring of gestationally stressed primate mothers.


Asunto(s)
Aminas Biogénicas/metabolismo , Encéfalo/metabolismo , Complicaciones del Embarazo/psicología , Efectos Tardíos de la Exposición Prenatal , Aislamiento Social , Estrés Psicológico , Ácido 3,4-Dihidroxifenilacético/metabolismo , Envejecimiento/metabolismo , Animales , Conducta Animal , Aminas Biogénicas/líquido cefalorraquídeo , Encéfalo/crecimiento & desarrollo , Femenino , Ácido Homovanílico/metabolismo , Ácido Hidroxiindolacético/metabolismo , Macaca mulatta , Masculino , Metoxihidroxifenilglicol/metabolismo , Ruido , Norepinefrina/metabolismo , Embarazo
16.
Am J Occup Ther ; 52(2): 90-8, 1998 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-9494629

RESUMEN

OBJECTIVES: The purpose of this study was to investigate whether maternal endocrine activation during pregnancy would affect the neurobehavioral state of primate offspring in a manner similar to that observed in human infants from pregnancies involving maternal substance abuse or maternal stress. METHOD: Twenty-two rhesus monkey (Macaca mulatta) infants were derived from females administered either adrenocorticotrophic hormone (ACTH), which increased the mother's endocrine activity, or saline solutions for 14 consecutive days during mid-pregnancy. On days 15 and 30 postpartum, infants underwent brief separations from their mothers and were videotaped for later evaluation of neurobehavioral state. RESULTS: Infants from mothers administered ACTH spent significantly more time in a drowsy state than did controls (p < .04), and the increased drowsiness tended to be most pronounced during the postseparation period, when acute stress was highest. In contrast, controls remained in a more active alert state (p < .03), presumably searching for their mother, a species-typical adaptive response to maternal separation. Female infants spent more time in distressed state than did males on day 15, and the proportion of time in distressed state decreased in all infants after administration of .2 ml of 24% sucrose solution. CONCLUSION: The results demonstrate that neurobehavioral state alterations are found in infants from mothers with increased endocrine activity during pregnancy. Neurobehavioral state disorganization can have an adverse impact on the human infant's concurrent and subsequent occupational performance. These findings establish the usefulness of the nonhuman primate model for advancing knowledge on early contributions to the development of human infant occupational behavior.


Asunto(s)
Hormona Adrenocorticotrópica/farmacología , Conducta Animal/efectos de los fármacos , Sistema Endocrino/efectos de los fármacos , Preñez/efectos de los fármacos , Hormona Adrenocorticotrópica/administración & dosificación , Animales , Animales Recién Nacidos , Ansiedad de Separación , Sistema Endocrino/metabolismo , Femenino , Humanos , Macaca mulatta , Masculino , Modelos Biológicos , Actividad Motora/efectos de los fármacos , Embarazo , Preñez/metabolismo , Efectos Tardíos de la Exposición Prenatal , Estrés Psicológico
18.
Biochemistry ; 34(51): 16574-84, 1995 Dec 26.
Artículo en Inglés | MEDLINE | ID: mdl-8527430

RESUMEN

Human erythrocyte band 3 inhibits glycolytic enzymes, including aldolase, by binding these cytoplasmic enzymes at its N-terminus. Phosphorylation of Y8 disrupts inhibition, and there is evidence that in vivo glycolysis levels in erythrocytes are regulated in part by a phosphorylation/dephosphorylation signaling pathway. The structural basis for control by phosphorylation has been investigated by NMR studies on a complex between aldolase and a synthetic peptide corresponding to the first 15 residues of band 3 (MEELQDDYEDMMEEN-NH2). The structure of this band 3 peptide (B3P) when it is bound to rabbit muscle aldolase was determined using the exchange-transferred nuclear Overhauser effect (ETNOE). Two hundred NMR structures for B3P were generated by simulated annealing molecular dynamics with NMR-derived distance restraints and excluding electrostatic terms. Twenty structures were further refined against a force field including full partial charges. The important conformational feature of B3P in the bound state is a folded loop structure involving residues 4-9 and M12 that surrounds Y8 and is stabilized by a hydrophobic cluster with the ring of Y8 sandwiched between the methyl groups of L4 and M12. Differential line broadening indicates that this loop structure binds aldolase in a relatively specific manner, while terminal regions are structurally heterogeneous. To better understand B3P inhibition of aldolase and the mechanism of phosphorylation control, a complex was modeled by docking B3P into the active site of aldolase and optimizing the fit using restrained molecular dynamics and energy minimization. The B3P loop is complementary in conformation to the beta-barrel central core containing the aldolase active site residues. Binding is electrostatic in nature with numerous ionic and hydrogen-bonding interactions involving several conserved lysine and arginine residues of aldolase. How phosphorylation of band 3 could disrupt inhibition was considered by modeling a phosphoryl moiety onto Y8 of B3P. An energetic analysis with respect to rigid phosphate rotation suggests that aldolase inhibition is reversed primarily because of electrostatic repulsion between B3P residues that destabilizes the B3P loop formed in the complex. This proposed intramolecular mechanism for blocking protein--protein association by electrostatic repulsion with the phosphoryl group may be applicable to other protein--protein signaling complexes.


Asunto(s)
Proteína 1 de Intercambio de Anión de Eritrocito/antagonistas & inhibidores , Proteína 1 de Intercambio de Anión de Eritrocito/química , Fructosa-Bifosfato Aldolasa/química , Secuencia de Aminoácidos , Animales , Proteína 1 de Intercambio de Anión de Eritrocito/genética , Sitios de Unión , Fructosa-Bifosfato Aldolasa/metabolismo , Humanos , Técnicas In Vitro , Cinética , Espectroscopía de Resonancia Magnética , Modelos Moleculares , Datos de Secuencia Molecular , Estructura Molecular , Fragmentos de Péptidos/química , Fragmentos de Péptidos/genética , Fragmentos de Péptidos/metabolismo , Fosforilación , Unión Proteica , Conformación Proteica , Conejos , Soluciones , Termodinámica , Tirosina/química
19.
Am J Otol ; 16(5): 687-9, 1995 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-8588678

RESUMEN

Implantable permanent magnets are increasingly used in devices for otolaryngologic applications. It is likely that at least some of the patients with implanted magnets will be in need of magnetic resonance imaging (MRI). The effect of an MRI scan on the magnetic properties of implanted permanent magnets has not been previously demonstrated. Some of the basic concepts and descriptive terminology used in industry regarding permanent magnets are reviewed. Experiments presented show that the MRI scan is capable of demagnetizing permanent magnets. A case history is also presented that demonstrates demagnetizing of an implanted Audiant magnet by an MRI scan.


Asunto(s)
Audífonos , Imagen por Resonancia Magnética/efectos adversos , Magnetismo , Pérdida Auditiva Sensorineural/terapia , Humanos , Masculino , Persona de Mediana Edad , Prótesis e Implantes
20.
Lab Anim Sci ; 44(4): 351-7, 1994 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-7983847

RESUMEN

Anecdotal evidence has accumulated from research and animal care personnel regarding the aggressive behavior reported in captive rhesus macaques originating in China. In this study, we compared neonatal temperament, activity, and neuromotor reflexes in 13 Chinese-Indian hybrid and 29 Indian-derived nursery-reared infants. Neonatal assessments were conducted on days 7, 14, 21, and 30, using a test based on the Brazelton Neonatal Assessment Scale developed for use in human newborns. Hybrid infants had lower scores for all items pertaining to orientation and ability to sustain attention. They were also rated as more temperamentally reactive and irritable than the Indian-derived infants. These results suggest that constitutional factors may underlie some of the behavioral differences observed in Chinese- and Indian-origin adults and that these qualities emerge very early in life.


Asunto(s)
Animales Recién Nacidos/fisiología , Conducta Animal , Macaca mulatta/fisiología , Temperamento , Agresión , Animales , China , Hibridación Genética , India , Actividad Motora
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