Effect of interfacial and edge roughness on magnetoelectric control of Co/Ni microdisks on PMN-PT(011).

Uniform magnetic behavior within arrays of magnetoelectric heterostructures is important for the development of reliable strain-mediated microdevices. Multiple mechanisms may contribute to observed nonuniform magnetization reversal including surface roughness, non-uniform strain, and fabrication induced imperfections. Here, Co/Ni microdisks of 7 µm diameter were produced on both [Pb(Mg1/3Nb2/3)O3]1-x-[PbTiO3]x with x = 0.3 nominal composition (PMN-30PT) (011) and Si substrates, and the out-of-plane magnetization reversal was characterized using magneto-optical Kerr effect (MOKE). Coercivity variation across the microdisks within the arrays was observed on both the PMN-30PT and Si specimens with zero electric field applied. Co/Ni microdisks on a PMN-30PT substrate displayed relatively larger coercivity than those on a Si substrate due to the surface roughness effect. Quasistatic electric fields of varying magnitude were applied to the PMN-30PT substrate to assess the dependence of the coercivity on electric field induced strain. Our results indicate that while coercivity decreases with the increase of electric field induced strain, interfacial and edge roughness combine to realize a prohibitively large coercivity to overcome within the Co/Ni microdisks as well as a broad distribution of coercive field across a patterned microdisk array.

Effect of CoFe dusting layer and annealing on the magnetic properties of sputtered Ta/W/CoFeB/CoFe/MgO layer structures.

We explored the effect of a CoFe wedge inserted as a dusting layer (0.2 nm-0.4 nm thick) at the CoFeB/MgO interface of a sputtered Ta(2 nm)/W(3 nm)/CoFeB(0.9 nm)/MgO(3 nm)/Ta(2 nm) film-a typical structure for spin-orbit torque devices. Films were annealed at temperatures varying between 300 °C and 400 °C in an argon environment. Ferromagnetic resonance studies and vibrating sample magnetometry measurements were carried out to estimate the effective anisotropy field, the Gilbert damping, the saturation magnetization and the dead layer thickness as a function of the CoFe thickness and across several annealing temperatures. While the as-deposited films present only easy-plane anisotropy, a transition along the wedge from in-plane to out-of-plane was observed across several annealing temperatures, with evidence of a spin-reorientation transition separating the two regions.

Influence of internal geometry on magnetization reversal in asymmetric permalloy rings.

We report the magnetization reversal behavior of microstructured Ni80Fe20 rings using magneto-optic indicator film imaging and magnetometry. While the reversal behavior of rings with a symmetric (circular) interior hole agrees with micromagnetic simulations of an onion → vortex → onion transition, we experimentally demonstrate that rings possessing an elliptical hole with an aspect ratio of 2 exhibit complex reversal behavior comprising incoherent domain propagation in the rings. Magneto optic images reveal metastable magnetic configurations that illustrate this incoherent behavior. These results have important implications for understanding the reversal behavior of asymmetric ferromagnetic rings.

Dihydrotestosterone and estrogen regulation of rat brain androgen-receptor immunoreactivity.

Androgen-receptor upregulation that occurs with androgenic-anabolic steroid (AAS) administration may be mediated by AAS metabolites, dihydrotestosterone (DHT), and estrogen. Castrated and intact male rats received 14 s.c. daily injections of AAS (2 mg/kg testosterone cypionate, 2 mg/kg nandrolone decanoate, and 1 mg/kg boldenone undecylenate in sesame oil vehicle), DHT (5 mg/kg dihydrotestosterone), EB (5 mg/kg estradiol benzoate), or sesame oil vehicle. Approximately 18-24 h after the fourteenth injection, brain tissues were removed and processed immunocytochemically using the PG-21 androgen-receptor antibody. As reported before, castration eliminated AR-ir (androgen-receptor immunoreactivity) and AAS upregulated AR-ir in the ventromedial hypothalamus (VMHVL), medial amygdala (MePV), and medial preoptic area (MPOM). When compared to AAS, DHT fully upregulated AR-ir in the VM VL and MPOM and partially upregulated AR-ir in the MePV. EB treatment partially upregulated AR-ir in the VMHVL and MePV, but not in the MPOM of castrated rats. Because AR-ir in the MPOM was consistently upregulated by DHT or AAS, and not EB, androgen-receptor availability in this region may be mediated specifically via androgen receptors.

Androgenic-anabolic steroids blunt morphine-induced c-fos expression in the rat striatum: possible role of beta-endorphin.

Self-administration of large doses of androgenic-anabolic steroids (AAS) in a significant portion of the population suggests that these agents are drugs of abuse. However, acute administration of AAS did not induce striatal immediate-early genes (IEG) expression in male rats, indicating that AAS do not share a common mechanism of action with other drugs of abuse. Surveys have indicated that people who abuse AAS are more likely to self-administer other drugs of abuse than do people who do not take AAS. In the present study, chronic administration of AAS blunted the striatal c-fos response to morphine, indicating that AAS can alter the molecular responses to at least one drug of abuse. Chronic administration of AAS also increased the content of beta-endorphin in the midline thalamus, suggesting a possible mechanism by which AAS may modulate the response to morphine through regulation of thalamo-striatal neurons.

Scopolamine inhibition of female sexual behavior in rhesus monkeys (Macaca mulatta).

Previous research supports the activational role of central cholinergic mechanisms in rodent female sexual behavior. This experiment examined if similar central cholinergic mechanisms facilitate female rhesus monkey (Macaca mulatta) sexual behavior. Eight ovariectomized female rhesus monkeys received daily estradiol benzoate priming (5 microg/kg, SC). After 13-16 days of estrogen priming, animals were injected intravenously with either the cholinergic antagonist, scopolamine (0.70 mg/kg), or saline vehicle (1 ml/kg). Results indicate that the female proceptive behaviors of noncontact presentations significantly decreased 1545 min after scopolamine injection. Scopolamine inhibition was sustained up to 75 min only after 15 days of estrogen priming. Scopolamine did not significantly reduce other female sexual behaviors. Additionally, significant decreases in the number of mounts and intromissions, but not hip touches, were displayed by males exposed to scopolamine-treated females. This research suggests the possibility of a central cholinergic mechanism regulating female sexual behavior in rhesus monkeys. However, the general nature and duration of the cholinergic regulation of primate female sexual behavior differs substantially when compared to rodent behavior.

BpaB25 insulins. Photoactivatable analogues that quantitatively cross-link, radiolabel, and activate the insulin receptor.

Benzoylphenylalanine (Bpa), a photoactivatable amino acid was incorporated into the 25 position of the insulin B-chain by a combination of chemical synthesis and enzymatic semisynthesis. BpaB25 insulin binds specifically to the insulin receptor with an affinity approximately 40% that of native insulin. Addition of biotin or epsilon-amino-hexanoic acid-iminobiotin at the epsilon-LysB29 position to form BpaB25,B29 epsilon-biotinyl (BBpa) or BpaB25,B29 epsilon-(Aha-iminobiotin) analogues had little adverse effect on receptor binding affinity and provided a convenient handle for affinity purification, gel shift assays, and blotting of cross-linked complexes with avidin. The analogues were readily 125I-iodinated with the majority of 125I being incorporated at the TyrA14 position; the monoiodo-A14 derivative was easily separated from other forms by high performance liquid chromatography. Photolysis of a complex of the insulin receptor and either BpaB25 insulin or [125I]iodo-TyrA14,BpaB25 insulin yields a covalent insulin-receptor complex. The efficiency of cross-linking with these reagents was unusually high, ranging from 60 to 100%. Furthermore, cross-linking to the insulin receptor results in kinase activation in vitro, and in intact cells insulin receptor phosphorylation and internalization were both activated. Notably, even at saturating concentrations one molecule of BpaB25 insulin covalently cross-linked each alpha 2 beta 2 receptor, demonstrating that holoreceptor activation occurs with one high affinity insulin binding site occupied and, if a second insulin binds with lower affinity it must be in a different orientation. Bpa insulin analogues form a new class of photoaffinity reagents which facilitate studies relating insulin-insulin receptor structure and function.

Mutations at the dimer, hexamer, and receptor-binding surfaces of insulin independently affect insulin-insulin and insulin-receptor interactions.

Mutagenesis of the dimer- and hexamer-forming surfaces of insulin yields analogues with reduced tendencies to aggregate and dramatically altered pharmacokinetic properties. We recently showed that one such analogue, HisB10----Asp, ProB28----Lys, LysB29----Pro human insulin (DKP-insulin), has enhanced affinity for the insulin receptor and is useful for studying the structure of the insulin monomer under physiologic solvent conditions [Weiss, M. A., Hua, Q. X., Lynch, C. S., Frank, B. H., & Shoelson, S. E. (1991) Biochemistry 30, 7373-7389]. DKP-insulin retains native secondary and tertiary structure in solution and may therefore provide an appropriate baseline for further studies of related analogues containing additional substitutions within the receptor-binding surface of insulin. To test this, we prepared a family of DKP analogues having potency-altering substitutions at the B24 and B25 positions using a streamlined approach to enzymatic semisynthesis which negates the need for amino-group protection. For comparison, similar analogues of native human insulin were prepared by standard semisynthetic methods. The DKP analogues show a reduced tendency to self-associate, as indicated by 1H-NMR resonance line widths. In addition, CD spectra indicate that (with one exception) the native insulin fold is retained in each analogue; the exception, PheB24----Gly, induces similar perturbations in both native insulin and DKP-insulin backgrounds. Notably, analogous substitutions exhibit parallel trends in receptor-binding potency over a wide range of affinities: D-PheB24 greater than unsubstituted greater than GlyB24 greater than SerB24 greater than AlaB25 greater than LeuB25 greater than SerB25, whether the substitution was in a native human or DKP-insulin background. Such "template independence" reflects an absence of functional interactions between the B24 and B25 sites and additional substitutions in DKP-insulin and demonstrates that mutations in discrete surfaces of insulin have independent effects on protein structure and function. In particular, the respective receptor-recognition (PheB24, PheB25), hexamer-forming (HisB10), and dimer-forming (ProB28, LysB29) surfaces of insulin may be regarded as independent targets for protein design. DKP-insulin provides an appropriate biophysical model for defining structure-function relationships in a monomeric template.

Heteronuclear 2D NMR studies of an engineered insulin monomer: assignment and characterization of the receptor-binding surface by selective 2H and 13C labeling with application to protein design.

Insulin provides an important model for the application of genetic engineering to rational protein design and has been well characterized in the crystal state. However, self-association of insulin in solution has precluded complementary 2D NMR study under physiological conditions. We demonstrate here that such limitations may be circumvented by the use of a monomeric analogue that contains three amino acid substitutions on the protein surface (HisB10----Asp, ProB28----Lys, and LysB29----Pro); this analogue (designated DKP-insulin) retains native receptor-binding potency. Comparative 1H NMR studies of native human insulin and a series of three related analogues--(i) the singly substituted analogue [HisB10----Asp], (ii) the doubly substituted analogue [ProB28----Lys; LysB29----Pro], and (iii) DKP-insulin--demonstrate progressive reduction in concentration-dependent line-broadening in accord with the results of analytical ultracentrifugation. Extensive nonlocal interactions are observed in the NOESY spectrum of DKP-insulin, indicating that this analogue adopts a compact and stably folded structure as a monomer in overall accord with crystal models. Site-specific 2H and 13C isotopic labels are introduced by semisynthesis as probes for the structure and dynamics of the receptor-binding surface. These studies confirm and extend under physiological conditions the results of a previous 2D NMR analysis of native insulin in 20% acetic acid [Hua, Q. X., & Weiss, M. A. (1991) Biochemistry 30, 5505-5515]. Implications for the role of protein flexibility in receptor recognition are discussed with application to the design of novel insulin analogues.