Ion-exchange in melanin: an electron spin resonance study with lanthanide probes.

Changes are induced in the electron spin resonance signal amplitude and microwave power saturation of the naturally occurring free radical in melanin by bound paramagnetic ions. The changes serve as experimental observables in competition experiments between diamagnetic and paramagnetic metal ions for melanin binding sites and between melanin and ethylenediaminetetraacetic acid for paramagnetic metal ions. Evidence is presented for the existence of several specific types of metal binding sites. The interaction of copper with free radicals leading to loss of electron spin resonance signal amplitude is magnetic in nature and not, as has been supposed, chemical.

Eumelanins and pheomelanins: characterization by electron spin resonance spectroscopy.

Synthetic dopa melanin and cysteinyldopa melanin have different electron spin resonance spectra. Data are reported for mixtures of these melanins and for dopa-cysteinyldopa copolymers, which are spectroscopically similar. A simple parameterization of the spectra allows estimation of the relative amounts of (i) dopa melanin and cysteinyldopa melanin in mixtures and of (ii) dopa and cysteinyldopa incorporated into copolymers. Several natural eumelanins and pheomelanins have been characterized and shown to be copolymers.

Coupling of Waveguide and Resonator by Inductive and Capacitive Irises for EPR Spectroscopy.

An analytic circuit model for slot coupling from a waveguide to a loop-gap resonator (LGR) in a context of electron paramagnetic resonance (EPR) spectroscopy is presented. The physical dimensions of the waveguide, iris, LGR, and aqueous sample are transformed into circuit values of inductance, capacitance, and resistance. These values are used in a solution of circuit equations that results in a prediction of the rf currents, magnitude and phase, frequency, and magnetic and electric stored energies near critical coupling. The circuit geometry reflects magnetic flux conservation between the iris and LGR as well as modification of the outer loop LGR currents by the iris. Unlike conventional models, coupling is not explicitly based on a mutual inductance between the iris and LGR. Instead, the conducting wall high frequency rf boundary condition is used to define surface currents, regions, and circuit topology with lumped-circuit values of self-inductance, capacitance, and resistance. Match is produced by a combination of self-inductive and capacitive circuit coupling. Two conditions must be met to achieve match. First, the equivalent resistance of the LGR as seen by the iris must be transformed into the waveguide characteristic impedance. This transformation is met at a particular frequency relative to the natural LGR resonance frequency. The frequency shift magnitude is largely determined by the LGR properties, weakly dependent on iris length and placement, and independent of other iris dimensions. The second condition for match is that the iris reactance at this frequency shift must cancel the residual reactance of the LGR. This second condition is sensitive to the iris dimensions. If both conditions are not simultaneously satisfied, overcoupling or undercoupling results. A slotted iris of equal length to the size of the large dimension of the waveguide is found to have many properties opposite to a conventional iris of shorter length. Notably, the magnetic field near the iris tends to reinforce rather than oppose the magnetic field in the resonator. The long iris improves the LGR EPR performance by providing increased rf magnetic field homogeneity at the sample, higher signal, and reduced total frequency shift since the shifts due to sample and iris tend to cancel. Investigations reveal that the first match condition can be adjusted by LGR dimensional changes and such adjustment can eliminate the frequency shift. Results are consistent with Ansoft High Frequency Structure Simulator (Version 10.1, Ansoft Corporation, Pittsburgh, PA) simulations and can be extended to cavity resonators.

Meta-metallic coils and resonators: Methods for high Q-value resonant geometries.

A novel method of decreasing ohmic losses and increasing Q-value in metallic resonators at high frequencies is presented. The method overcomes the skin-depth limitation of rf current flow cross section. The method uses layers of conductive foil of thickness less than a skin depth and capacitive gaps between layers. The capacitive gaps can substantially equalize the rf current flowing in each layer, resulting in a total cross-sectional dimension for rf current flow many times larger than a skin depth. Analytic theory and finite-element simulations indicate that, for a variety of structures, the Q-value enhancement over a single thick conductor approaches the ratio of total conductor thickness to skin depth if the total number of layers is greater than one-third the square of the ratio of total conductor thickness to skin depth. The layer number requirement is due to counter-currents in each foil layer caused by the surrounding rf magnetic fields. We call structures that exhibit this type of Q-enhancement "meta-metallic." In addition, end effects due to rf magnetic fields wrapping around the ends of the foils can substantially reduce the Q-value for some classes of structures. Foil structures with Q-values that are substantially influenced by such end effects are discussed as are five classes of structures that are not. We focus particularly on 400 MHz, which is the resonant frequency of protons at 9.4 T. Simulations at 400 MHz are shown with comparison to measurements on fabricated structures. The methods and geometries described here are general for magnetic resonance and can be used at frequencies much higher than 400 MHz.

Electron-electron double resonance measurements on xanthine oxidase.

Electron-electron double resonance measurements were carried out on milk xanthine oxidase (xanthine:oxygen oxidoreductase EC 1.2.3.2) and the spectra obtained supported a previous model, based on EPR data, proposing a spin-spin interaction between unpaired electrons associated with Fe-S and Mo. The technique demonstrated that the additional apparently isotropic, splitting in the Mo EPR spectra observed at low temperature is produced by a single site giving two spectra interconverting at a rate consistent with the Fe-S spin lattice relaxation time. Other data concerning the model and the relaxation behaviour of the species are discussed.

The diffusion-concentration product of oxygen in lipid bilayers using the spin-label T1 method.

A method is described to measure the oxygen diffusion-concentration product, DO[O2], at any locus that can be probed or labeled using nitroxide radicals. The method is based on the dependence of the spin-lattice relaxation time T1 of the spin label on the bimolecular collision rate with oxygen. Strong Heisenberg exchange between spin label and oxygen contributes directly to T1 of the spin label, while dipolar interactions are negligible. Both time-domain and continuous wave saturation methods for studying T1 are considered. The method has been applied to phospholipid liposomes using fatty acid spin labels. A discontinuity in DO[O2] at the main phase transition was observed.

Depth dependence of the perturbing effect of placing a bulky group (oxazolidine ring spin labels) in the membrane on the membrane phase transition.

Electron paramagnetic resonance (EPR) and differential scanning calorimetry (DSC) have been used to study the effect on the phase transition of dimyristoylphosphatidylcholine membranes of incorporating various stearic acid spin labels (SASL's) that contain the bulky oxazolidine ring at various positions along the stearyl chain. SASL's lowered the phase transition temperature and decreased the size of the cooperative unit, with the effects stronger in the order of 9- > 12- > 5- > 16-SASL > stearic acid (no label). Incorporation of stearic acid without the spin label slightly increases the phase transition temperature. Incorporation of 9-SASL (3 mol% of lipid) decreased the transition temperature by 1.8 degrees C and the cooperative unit to 1/5 of that without the spin label, while the effect of 16-SASL was slight. The effect on transition enthalpy was small. It is concluded that the perturbing effect of placing a bulky group on the alkyl chain on phase transition is through inducing packing defects in the gel-phase.

Oxygen permeability of thylakoid membranes: electron paramagnetic resonance spin labeling study.

Oxygen transport in thylakoid membranes of spinach chloroplasts (Spinacia oleracea) has been studied by observing the collisions of molecular oxygen with spin labels, using line broadening electron paramagnetic resonance (EPR) spectroscopy. Stearic acid spin labels were used to probe the local oxygen diffusion-concentration product. The free radical moiety was located at various distances from the membrane surface, and collision rates were estimated from linewidths of the EPR spectra measured in the presence and absence of molecular oxygen. The profile of the local oxygen diffusion-concentration product across the membrane determined at 20 degrees C demonstrates that this product, at all membrane locations, is higher than the value measured in water. From the profile of the oxygen diffusion-concentration product, the membrane oxygen permeability coefficient has been estimated using the procedure developed earlier (W.K. Subczynski, J.S. Hyde, A. Kusumi, Proc. Natl. Acad. Sci. USA 86 (1989) 4474-4478). At 20 degrees C, the oxygen permeability coefficient for the lipid portion of the thylakoid membrane was found to be 39.5 cm s-1. This value is 20% higher than the oxygen permeability coefficient of a water layer of the same thickness as the thylakoid membrane. The high permeability coefficient implies that the oxygen concentration difference across the thylakoid membrane generated under the illumination of the leaf by saturating actinic light is negligible, smaller than 1 microM.

Spin-label studies on phosphatidylcholine-cholesterol membranes: effects of alkyl chain length and unsaturation in the fluid phase.

Dynamic properties of phosphatidylcholine-cholesterol membranes in the fluid phase and water accessibility to the membranes have been studied as a function of phospholipid alkyl chain length, saturation, mole fraction of cholesterol, and temperature by using spin and fluorescence labelling methods. The results are the following: (1) The effect of cholesterol on motional freedom of 5-doxyl stearic acid spin label (5-SASL) and 16-doxyl stearic acid spin label (16-SASL) in saturated phosphatidylcholine membrane is significantly larger than the effects of alkyl chain length and introduction of unsaturation in the alkyl chain. (2) Variation of alkyl chain length of saturated phospholipids does not alter the effects of cholesterol except in the case of dilauroylphosphatidylcholine, which possesses the shortest alkyl chains (12 carbons) used in this work. (3) Unsaturation of the alkyl chains greatly reduces the ordering effect of cholesterol at C-5 and C-16 positions although unsaturation alone gives only minor fluidizing effects. (4) Introduction of 30 mol% cholesterol to dimyristoylphosphatidylcholine membranes decreases the lateral diffusion constants of lipids by a factor of four, while it causes only a slight decrease of lateral diffusion in dioleoylphosphatidylcholine membranes. (5) If compared at the same temperature, 5-SASL mobilities plotted as a function of mole fraction of cholesterol in the fluid phases of dimyristoylphosphatidylcholine-, dipalmitoylphosphatidylcholine- and distearoylphosphatidylcholine-cholesterol membranes are similar in wide ranges of temperature (45-82 degrees C) and cholesterol mole fraction (0-50%). (6) In isothermal experiments with saturated phosphatidylcholine membranes, 5-SASL is maximally immobilized at the phase boundary between Regions I and III reported by other workers (Recktenwald, D.J. and McConnell, H.M. (1981) Biochemistry 20, 4505-4510) and becomes more mobile away from the boundary in Regions I and III. (7) 5-SASL in unsaturated phosphatidylcholine membranes showed a gradual monotonic immobilization with increase of cholesterol mole fraction without showing any maximum in the range of cholesterol fractions studied. (8) By rigorously determining rigid-limit magnetic parameters of cholestane spin labels in membranes from Q-band second-derivative ESR spectra to monitor the dielectric environment around the nitroxide radical, it is concluded that cholesterol incorporation increases water accessibility in the hydrophilic loci of the membrane. In contrast, 12-(9-anthroyloxy)stearic acid fluorescence showed that water accessibility is decreased in the hydrophobic loci of the membrane.

Stopped-flow kinetic and biophysical studies of membrane-associated D-lactate dehydrogenase of Escherichia coli.

The enzyme kinetics of the FAD-containing membrane-associated D-lactate dehydrogenase (D-LDH) of Escherichia coli have been investigated by stopped-flow spectroscopy. The reduction of D-LDH by the substrate, D-lactate, exhibits a two-stage behavior as observed by the absorbance change for the enzyme-bound FAD. The fast stage with a maximum rate of 400 s-1 represents the rapid formation of the enzyme-substrate complex and the formation of the equilibrium between the oxidized and the reduced enzyme-substrate complexes. The slow stage, which occurs on the order of 0.36 s-1, represents the slow release of the product, pyruvate, from the reduced enzyme. The formation of a D-LDH semiquinone radical was not observed during the oxidation of D-lactate by D-LDH at 25 degrees C. However, during the subsequent electron transfer from the reduced enzyme to a nitroxide spin-label, a one-electron acceptor, an enzyme intermediate has been observed and identified by both optical and EPR spectroscopies as an anionic semiquinone. Results from 1H-NMR spectroscopic studies suggest the possible formation of a substrate carbanion when D-lactate is bound at the active site of D-LDH.

Is the mammalian cell plasma membrane a barrier to oxygen transport?

Oxygen transport in the Chinese hamster ovary (CHO) plasma membrane has been studied by observing the collision of molecular oxygen with nitroxide radical spin labels placed in the lipid bilayer portion of the membrane at various distances from the membrane surface using the long-pulse saturation-recovery electron spin resonance (ESR) technique. The collision rate was estimated for 5-, 12-, and 16-doxylstearic acids from spin-lattice relaxation times (T1) measured in the presence and absence of molecular oxygen. Profiles of the local oxygen transport parameters across the membrane were obtained showing that the oxygen diffusion-concentration product is lower than in water for all locations at 37 degrees C. From oxygen transport parameter profiles, the membrane oxygen permeability coefficients were estimated according to the procedure developed earlier by Subczynski et al. (Subczynski, W. K., J. S. Hyde, and A. Kusumi. 1989. Proceedings of the National Academy of Sciences, USA. 86:4474-4478). At 37 degrees C, the oxygen permeability coefficient for the plasma membrane was found to be 42 cm/s, about two times lower than for a water layer of the same thickness as the membrane. The oxygen concentration difference across the CHO plasma membrane at physiological conditions is in the nanomolar range. It is concluded that oxygen permeation across the cell plasma membrane cannot be a rate-limiting step for cellular respiration. Correlations of the form PM = cKs between membrane permeabilities PM of small nonelectrolyte solutes of mol wt less than 50, including oxygen, and their partition coefficients K into hexadecane and olive oil are reported. Hexadecane: c = 26 cm/s, s = 0.95; olive oil: c = 23 cm/s, s = 1.56. These values of c and s differ from those reported in the literature for solutes of 50 less than mol wt less than 300 (Walter, A., and J. Gutknecht. 1986. Journal of Membrane Biology. 90:207-217). It is concluded that oxygen permeability through membranes can be reliably predicted from measurement of partition coefficients.

Hypercapnia reversibly suppresses low-frequency fluctuations in the human motor cortex during rest using echo-planar MRI.

Using magnetic resonance (MR) echo-planar imaging (EPI), we recently demonstrated the presence of low-frequency fluctuations (< 0.1 Hz) in MR signal intensity from the resting human brain that have a high degree of temporal correlation (p < 10(-3)) within and across associated regions of the sensorimotor cortex. These fluctuations in MR signal intensity are believed to arise from fluctuations in capillary blood flow and oxygenation. A substantial overlap between the activation map generated by bilateral finger tapping and temporally-correlated voxels from the sensorimotor cortex obtained during rest was observed. In the work reported here, we investigated whether respiratory hypercapnia, which is known to suspend spontaneous oscillations in regional cerebral blood flow, influences these low-frequency fluctuations. The magnitude of low-frequency fluctuations was reversibly diminished during hypercapnia, resulting in a substantial decrease of the temporal correlation both within and across contralateral hemispheres of the sensorimotor cortex. After the breathing mixture was returned to ambient air, the magnitude and spatial extent of the temporal correlation of low-frequency fluctuations returned to normal. Results of this study support the hypothesis that low-frequency physiological fluctuations observed by MR in the human cortex and spontaneous flow oscillations observed in early studies by laser-Doppler flowmetry (LDF) in the cortex of the rat are identical and are secondary to fluctuations in neuronal activity.

Relationship between finger movement rate and functional magnetic resonance signal change in human primary motor cortex.

Functional magnetic resonance imaging (FMRI) is a noninvasive technique for mapping regional brain changes in response to sensory, motor, or cognitive activation tasks. Interpretation of these activation experiments may be confounded by more elementary task parameters, such as stimulus presentation or movement rates. We examined the effect of movement rate on the FMRI response recorded from the contralateral primary motor cortex. Four right-handed healthy subjects performed flexion-extension movements of digits 2-5 of the right hand at rates of 1, 2, 3, 4, or 5 Hz. Results of this study indicated a positive linear relationship between movement rate and FMRI signal change. Additionally, the number of voxels demonstrating functional activity increased significantly with faster movement rates. The magnitude of the signal change at each movement rate remained constant over the course of three 8-min scanning series. These findings are similar to those of previous rate studies of the visual and auditory system performed with positron emission tomography (PET) and FMRI.

Metaproterenol in children with chronic asthma.

Metaproterenol sulfate was given to 25 children with asthma in the form of a syrup in a continuous treatment course of 180 days. Dosage amounted to 10 to 20 mg metaproterenol four times daily, depending on the patient's age and weight. Double-blind crossover tests of pulmonary function were run against placebo at the beginning and after 3 and 6 mo of treatment. The peak expiratory flow rate responses to metaproterenol consistently exceeded the responses to placebo, the differences proving statistically significant at several intervals after administration. Adverse reactions were limited to instances of hyperactivity in one child and of mild tremors in another; laboratory values remained normal throughout the 6-mo period except for low normal fasting blood sugar values recorded in 2 patients at the end of the study. Pulse rate increases occurring after metaproterenol were not considered clinically important.

Lateralized human brain language systems demonstrated by task subtraction functional magnetic resonance imaging.

OBJECTIVE: To develop a procedure for noninvasive measurement of language lateralization with functional magnetic resonance imaging (MRI). DESIGN: Functional neuroimaging using time-series echo-planar MRI. SETTING: University medical center research facility. SUBJECTS: Five healthy, right-handed, young adults. MAIN OUTCOME MEASURES: Number of MRI voxels in left and right hemispheres showing task-related signal increases during two contrasting auditory processing tasks. The nonlinguistic task involved processing of pure tones, while the linguistic task involved processing of single words based on semantic content. RESULTS: The pure-tone processing task activated temporal lobe auditory areas and dorsolateral frontal regions bilaterally. Using this task as a control condition, the semantic processing task resulted in lateralized activity in distributed regions of the left hemisphere. A significant effect of task on intrahemispheric activity pattern was demonstrated in every subject. Results were reproduced in preliminary studies of test-retest reliability. CONCLUSIONS: The results demonstrate the lateralized anatomy of semantic linguistic systems in contrast to non-linguistic auditory sensory processors and introduce a task subtraction technique adapted for functional MRI as a noninvasive measure of language lateralization.

Somatotopic mapping of the human primary motor cortex with functional magnetic resonance imaging.

We applied functional magnetic resonance imaging (FMRI) to map the somatotopic organization of the primary motor cortex using voluntary movements of the hand, arm, and foot. Eight right-handed healthy subjects performed self-paced, repetitive, flexion/extension movements of the limbs while undergoing echo-planar imaging. Four subjects performed movements of the right fingers and toes, while the remaining subjects performed movements of the right fingers and elbow joint. There was statistically significant functional activity in the left primary motor cortex in all subjects. The pattern of functional activity followed a topographic representation: finger movements resulted in signal intensity changes over the convexity of the left motor cortex, whereas toe movements produced changes either at the interhemispheric fissure or on the dorsolateral surface adjacent to the interhemispheric fissure. Elbow movements overlapped the more medial signal intensity changes observed with finger movements. Functionally active regions were confined to the cortical ribbon and followed the gyral anatomy closely. These findings indicate that FMRI is capable of generating somatotopic maps of the primary motor cortex in individual subjects.

Gender differences in erotic plasticity--evolutionary or sociocultural forces? Comment on Baumeister (2000)

R. F. Baumeister (2000) argued that there are gender differences in erotic plasticity, meaning that women are more influenced by cultural and social factors than men are. He attributed the gender difference in erotic plasticity to evolutionary, biological forces. We propose an alternative account of the data using a multifactor sociocultural model that rests on 4 assertions: (a) Men have more power than women on many levels including the institutional and the interpersonal levels, (b) education increases women's power, (c) groups with less power (women) pay more attention to and adapt their behavior more to the group with more power (men) than the reverse, and (d) gender roles powerfully shape behavior, and heterosexuality is a more important element of the male role than the female role.

Gender differences in moral orientation: a meta-analysis.

C. Gilligan's (1982) critique of L. Kohlberg's theory of moral reasoning and her assertion that two modes of moral reasoning (justice and care) exist have been the subject of debate within the field of psychology for more than 15 years. This meta-analysis was conducted to review quantitatively the work on gender differences in moral orientation. The meta-analysis revealed small differences in the care orientation favoring females (d = -.28) and small differences in the justice orientation favoring males (d = .19). Together, the moderator variables accounted for 16% of the variance in the effect sizes for care reasoning and 17% of the variance in the effect sizes for justice reasoning. These findings do not offer strong support for the claim that the care orientation is used predominantly by women and that the justice orientation is used predominantly by men.

Gender differences in sexuality: a meta-analysis.

This meta-analysis surveyed 177 usable sources that reported data on gender differences on 21 different measures of sexual attitudes and behaviors. The largest gender difference was in incidence of masturbation: Men had the greater incidence (d = .96). There was also a large gender difference in attitudes toward casual sex: Males had considerably more permissive attitudes (d = .81). There were no gender differences in attitudes toward homosexuality or in sexual satisfaction. Most other gender differences were in the small-to-moderate range. Gender differences narrowed from the 1960s to the 1980s for many variables. Chodorow's neoanalytic theory, sociobiology, social learning theory, social role theory, and script theory are discussed in relation to these findings.