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1.
Sci Rep ; 10(1): 934, 2020 01 22.
Article in English | MEDLINE | ID: mdl-31969626

ABSTRACT

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.
Article in English | MEDLINE | ID: mdl-23208243

ABSTRACT

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).


Subject(s)
Dopamine Plasma Membrane Transport Proteins/analysis , Fluorine Radioisotopes/chemistry , Nortropanes/chemical synthesis , Chromatography, High Pressure Liquid
3.
Synapse ; 65(12): 1309-18, 2011 Dec.
Article in English | MEDLINE | ID: mdl-21674627

ABSTRACT

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.


Subject(s)
Pyridines/metabolism , Pyrroles/metabolism , Receptors, Nicotinic/metabolism , Animals , Binding Sites/physiology , Brain/diagnostic imaging , Brain/metabolism , Female , Macaca mulatta , Male , Neuroimaging/methods , Positron-Emission Tomography/methods , Protein Binding/physiology
4.
Synapse ; 65(7): 592-600, 2011 Jul.
Article in English | MEDLINE | ID: mdl-21484878

ABSTRACT

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).


Subject(s)
Brain/diagnostic imaging , Piperazines/pharmacokinetics , Pyridines/pharmacokinetics , Radiopharmaceuticals/pharmacokinetics , Animals , Carbon Radioisotopes/pharmacokinetics , Fluorine Radioisotopes/pharmacokinetics , Macaca mulatta , Positron-Emission Tomography/methods
5.
Neurotoxicol Teratol ; 26(2): 169-78, 2004.
Article in English | MEDLINE | ID: mdl-15019951

ABSTRACT

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.


Subject(s)
Behavior, Animal/drug effects , Central Nervous System Depressants/pharmacology , Dopamine/metabolism , Ethanol/pharmacology , Fetus/drug effects , Prenatal Exposure Delayed Effects , Stress, Psychological , Analysis of Variance , Animals , Animals, Newborn , Cognition/physiology , Female , Inhibition, Psychological , Macaca mulatta , Male , Motor Activity/drug effects , Pregnancy , Random Allocation , Stereotyped Behavior/drug effects , Tomography, Emission-Computed/methods
7.
Oncogene ; 20(51): 7524-35, 2001 Nov 08.
Article in English | MEDLINE | ID: mdl-11709724

ABSTRACT

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.


Subject(s)
Avian Proteins , Carrier Proteins/metabolism , Neoplasm Proteins/genetics , Neoplasm Proteins/metabolism , Nuclear Proteins/metabolism , Oncogene Protein p65(gag-jun)/metabolism , Amino Acid Sequence , Animals , Antibodies/metabolism , Base Sequence , Blotting, Northern , Carrier Proteins/chemistry , Cell Nucleolus/metabolism , Cell Transformation, Neoplastic , Chick Embryo , Chromatography , Cloning, Molecular , Coturnix , DNA/metabolism , DNA, Complementary/metabolism , Doxycycline/pharmacology , Enzyme Activation , Fibroblasts/metabolism , Humans , Kinetics , Mice , Models, Genetic , Molecular Sequence Data , Nuclear Proteins/chemistry , Precipitin Tests , Protein Binding , Protein Biosynthesis , Protein Structure, Tertiary , Proteins/metabolism , RNA/metabolism , Recombinant Proteins/metabolism , Retroviridae/genetics , Sequence Analysis, DNA , Sequence Homology, Amino Acid , Time Factors , Transcription, Genetic , Transcriptional Activation , Tumor Cells, Cultured
8.
Alcohol Clin Exp Res ; 25(9): 1383-92, 2001 Sep.
Article in English | MEDLINE | ID: mdl-11584160

ABSTRACT

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.


Subject(s)
Behavior, Animal/drug effects , Ethanol/pharmacology , Learning/drug effects , Prenatal Exposure Delayed Effects , Animals , Ethanol/administration & dosage , Female , Impulsive Behavior , Irritable Mood , Macaca mulatta , Motor Activity , Pregnancy , Stereotyped Behavior , Stress, Physiological/complications
9.
Alcohol Clin Exp Res ; 25(8): 1238-45, 2001 Aug.
Article in English | MEDLINE | ID: mdl-11505056

ABSTRACT

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.


Subject(s)
Animals, Newborn , Ethanol/administration & dosage , Maternal-Fetal Exchange , Pregnancy Outcome , Animals , Behavior, Animal , Birth Weight , Female , Gestational Age , Macaca mulatta , Male , Maternal Behavior , Motor Activity , Pregnancy , Time Factors
10.
J Mol Biol ; 307(5): 1395-410, 2001 Apr 13.
Article in English | MEDLINE | ID: mdl-11292350

ABSTRACT

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.


Subject(s)
DNA/metabolism , Oncogene Protein p55(v-myc)/chemistry , Oncogene Protein p55(v-myc)/metabolism , Transcription Factors/chemistry , Transcription Factors/metabolism , Amino Acid Sequence , Animals , Basic-Leucine Zipper Transcription Factors , Chickens , Circular Dichroism , DNA/genetics , DNA-Binding Proteins/chemistry , DNA-Binding Proteins/genetics , DNA-Binding Proteins/metabolism , Dimerization , Entropy , Helix-Loop-Helix Motifs , Leucine Zippers , Magnetic Resonance Spectroscopy , Molecular Sequence Data , Motion , Oncogene Protein p55(v-myc)/genetics , Protein Denaturation , Protein Folding , Protein Structure, Secondary , Protein Structure, Tertiary , Recombinant Proteins/chemistry , Recombinant Proteins/metabolism , Sequence Alignment , Solubility , Structure-Activity Relationship , Temperature , Transcription Factors/genetics
11.
Mol Cell Neurosci ; 18(5): 458-72, 2001 Nov.
Article in English | MEDLINE | ID: mdl-11922138

ABSTRACT

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.


Subject(s)
Cell Differentiation/physiology , Central Nervous System/embryology , Eye Proteins/metabolism , Membrane Proteins/metabolism , Neurons/metabolism , Retina/embryology , Saccharomyces cerevisiae Proteins , Transcription Factors/metabolism , Xenopus Proteins , Xenopus laevis/embryology , Amino Acid Motifs/genetics , Animals , Basic Helix-Loop-Helix Transcription Factors , Binding Sites/genetics , Blastomeres/cytology , Blastomeres/drug effects , Blastomeres/metabolism , Cell Differentiation/drug effects , Central Nervous System/cytology , Central Nervous System/metabolism , DNA-Binding Proteins/genetics , Eye Proteins/genetics , Female , Fungal Proteins/genetics , Fungal Proteins/metabolism , Gene Expression Regulation, Developmental/physiology , Green Fluorescent Proteins , Intracellular Signaling Peptides and Proteins , Luminescent Proteins/genetics , Membrane Proteins/genetics , Neurons/cytology , Protein Serine-Threonine Kinases/genetics , Protein Serine-Threonine Kinases/metabolism , Protein-Tyrosine Kinases/genetics , Protein-Tyrosine Kinases/metabolism , RNA, Messenger/genetics , RNA, Messenger/pharmacology , Receptors, Notch , Retina/cytology , Retina/metabolism , Retinal Ganglion Cells/cytology , Retinal Ganglion Cells/metabolism , Transcription Factors/genetics , Tubulin/metabolism , Xenopus laevis/metabolism
12.
Stress ; 4(3): 183-93, 2001 Sep.
Article in English | MEDLINE | ID: mdl-22432139

ABSTRACT

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.


Subject(s)
Behavior, Animal , Learning , Maternal Behavior , Pregnancy Complications/etiology , Prenatal Exposure Delayed Effects , Stress, Psychological/complications , Age Factors , Animals , Anxiety, Separation/metabolism , Anxiety, Separation/psychology , Attention , Disease Models, Animal , Female , Gestational Age , Humans , Hypothalamo-Hypophyseal System/metabolism , Macaca mulatta , Motor Activity , Pituitary-Adrenal System/metabolism , Pregnancy , Pregnancy Complications/psychology , Stress, Psychological/psychology
13.
Med Res Rev ; 19(4): 295-305, 1999 Jul.
Article in English | MEDLINE | ID: mdl-10398926

ABSTRACT

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.


Subject(s)
Phosphotyrosine/chemistry , Phosphotyrosine/physiology , Protein Sorting Signals/chemistry , Protein Sorting Signals/physiology , Drug Design , Humans , Models, Structural , Protein Conformation
14.
Child Dev ; 70(2): 263-74, 1999.
Article in English | MEDLINE | ID: mdl-10218255

ABSTRACT

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.


Subject(s)
Animals, Newborn/growth & development , Animals, Suckling/growth & development , Developmental Disabilities/etiology , Maternal Exposure/adverse effects , Prenatal Exposure Delayed Effects , Stress, Psychological/physiopathology , Animals , Animals, Newborn/psychology , Animals, Suckling/psychology , Birth Weight , Female , Handling, Psychological , Hydrocortisone/blood , Macaca mulatta , Male , Motor Skills/physiology , Orientation/physiology , Pregnancy , Stress, Psychological/blood
15.
Dev Psychopathol ; 10(3): 427-40, 1998.
Article in English | MEDLINE | ID: mdl-9741675

ABSTRACT

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.


Subject(s)
Biogenic Amines/metabolism , Brain/metabolism , Pregnancy Complications/psychology , Prenatal Exposure Delayed Effects , Social Isolation , Stress, Psychological , 3,4-Dihydroxyphenylacetic Acid/metabolism , Aging/metabolism , Animals , Behavior, Animal , Biogenic Amines/cerebrospinal fluid , Brain/growth & development , Female , Homovanillic Acid/metabolism , Hydroxyindoleacetic Acid/metabolism , Macaca mulatta , Male , Methoxyhydroxyphenylglycol/metabolism , Noise , Norepinephrine/metabolism , Pregnancy
16.
Am J Occup Ther ; 52(2): 90-8, 1998 Feb.
Article in English | MEDLINE | ID: mdl-9494629

ABSTRACT

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.


Subject(s)
Adrenocorticotropic Hormone/pharmacology , Behavior, Animal/drug effects , Endocrine System/drug effects , Pregnancy, Animal/drug effects , Adrenocorticotropic Hormone/administration & dosage , Animals , Animals, Newborn , Anxiety, Separation , Endocrine System/metabolism , Female , Humans , Macaca mulatta , Male , Models, Biological , Motor Activity/drug effects , Pregnancy , Pregnancy, Animal/metabolism , Prenatal Exposure Delayed Effects , Stress, Psychological
18.
Biochemistry ; 34(51): 16574-84, 1995 Dec 26.
Article in English | MEDLINE | ID: mdl-8527430

ABSTRACT

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.


Subject(s)
Anion Exchange Protein 1, Erythrocyte/antagonists & inhibitors , Anion Exchange Protein 1, Erythrocyte/chemistry , Fructose-Bisphosphate Aldolase/chemistry , Amino Acid Sequence , Animals , Anion Exchange Protein 1, Erythrocyte/genetics , Binding Sites , Fructose-Bisphosphate Aldolase/metabolism , Humans , In Vitro Techniques , Kinetics , Magnetic Resonance Spectroscopy , Models, Molecular , Molecular Sequence Data , Molecular Structure , Peptide Fragments/chemistry , Peptide Fragments/genetics , Peptide Fragments/metabolism , Phosphorylation , Protein Binding , Protein Conformation , Rabbits , Solutions , Thermodynamics , Tyrosine/chemistry
19.
Am J Otol ; 16(5): 687-9, 1995 Sep.
Article in English | MEDLINE | ID: mdl-8588678

ABSTRACT

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.


Subject(s)
Hearing Aids , Magnetic Resonance Imaging/adverse effects , Magnetics , Hearing Loss, Sensorineural/therapy , Humans , Male , Middle Aged , Prostheses and Implants
20.
Lab Anim Sci ; 44(4): 351-7, 1994 Aug.
Article in English | MEDLINE | ID: mdl-7983847

ABSTRACT

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.


Subject(s)
Animals, Newborn/physiology , Behavior, Animal , Macaca mulatta/physiology , Temperament , Aggression , Animals , China , Hybridization, Genetic , India , Motor Activity
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