Site-Specific Structural Variations Accompanying Tubular Assembly of the HIV-1 Capsid Protein

The 231-residue capsid (CA) protein of human immunodeficiency virus type 1 (HIV-1) spontaneously self-assembles into tubes with a hexagonal lattice that is believed to mimic the surface lattice of conical capsid cores within intact virions. We report the results of solid-state nuclear magnetic resonance (NMR) measurements on HIV-1 CA tubes that provide new information regarding changes in molecular structure that accompany CA self-assembly, local dynamics within CA tubes, and possible mechanisms for the generation of lattice curvature. This information is contained in site-specific assignments of signals in two- and three-dimensional solid-state NMR spectra, conformation-dependent ¹⁵N and ¹³C NMR chemical shifts, detection of highly dynamic residues under solution NMR conditions, measurements of local variations in transverse spin relaxation rates of amide ¹H nuclei, and quantitative measurements of site-specific ¹⁵N–¹⁵N dipole–dipole couplings. Our data show that most of the CA sequence is conformationally ordered and relatively rigid in tubular assemblies and that structures of the N-terminal domain (NTD) and the C-terminal domain (CTD) observed in solution are largely retained. However, specific segments, including the N-terminal β-hairpin, the cyclophilin A binding loop, the inter-domain linker, segments involved in intermolecular NTD–CTD interactions, and the C-terminal tail, have substantial static or dynamical disorder in tubular assemblies. Other segments, including the 3₁₀-helical segment in CTD, undergo clear conformational changes. Structural variations associated with curvature of the CA lattice appear to be localized in the inter-domain linker and intermolecular NTD–CTD interface, while structural variations within NTD hexamers, around local 3-fold symmetry axes, and in CTD–CTD dimerization interfaces are less significant.     

Comparison of 60-Hz electric fields and incandescent light as aversive stimuli controlling the behavior of rats

Rats were exposed to two procedures which enabled them to press a lever to turn off a 90 or 100 kV/m 60-Hz electric field or, later in the study, illumination from an incandescent lamp. Under one procedure, a response turned off the stimulus for a fixed duration, after which the stimulus was turned on again. A response during the off-period restarted the fixed duration. None of the rats turned the field off reliably. Next, under an alternative procedure, pressing one lever turned the field off; pressing the other lever turned it back on; responding under those conditions differed little from that seen at 0 kV/m. Under both procedures, when illumination from an incandescent lamp served as the stimulus, each rat did turn the stimulus off, and performances varied with stimulus intensity. The results show that a 100 kV/m 60-Hz electric field is not sufficient to function as an aversive stimulus under two procedures where illumination from a lamp does function as an aversive stimulus.     

Chronic exposure to ELF fields may induce depression

Exposure to extremely-low-frequency (ELF) electric or magnetic fields has been postulated as a potentially contributing factor in depression. Epidemiologic studies have yielded positive correlations between magnetic- and/or electric-field strengths in local environments and the incidence of depression-related suicide. Chronic exposure to ELF electric or magnetic fields can disrupt normal circadian rhythms in rat pineal serotonin-N-acetyltransferase activity as well as in serotonin and melatonin concentrations. Such disruptions in the circadian rhythmicity of pineal melatonin secretion have been associated with certain depressive disorders in human beings. In the rat, ELF fields may interfere with tonic aspects of neuronal input to the pineal gland, giving rise to what may be termed "functional pinealectomy." If long-term exposure to ELF fields causes pineal dysfunction in human beings as it does in the rat, such dysfunction may contribute to the onset of depression or may exacerbate existing depressive disorders.     

Effect of electrical fields and external ionic currents on pollen-tube orientation

Summary Agapanthus umbelatus pollen tubes (PTs) display a number of different growth patterns when germinated in an electric field of 750 mV· mm^–1. When pollen is germinated near the cathode (82.44% of orientation to the cathode side) or near the anode (55.35% of orientation to the anode), growth is oriented parallel to the applied field but when germinated at an intermediate position, there is random growth. An increase and decrease in the orientation rates as well as reversion of the polarized growth were observed when the growth conditions were systematically altered. These findings reflect the influence of different ionic currents present in the germination medium. These ionic currents induce the formation of ionic gradients, which were monitored by ion-HPLC. The individual omission of Ca²⁺, K⁺, Mg²⁺ and Cl^– suppresses or alters the oriented growth pattern. The presence of ionic gradients is not by itself suficient to trigger the polarization of tube growth as the presence of an electric field which drives the ionic currents is essential for this to occur.Abbreviations PT Pollen tube - DNS 3,5-dinitro salycilic acid; - TP transient polarization - HPLC high precision liquid chroma tography - DC direct current     

Alkaline phosphatase activities of anAnabaena sp. from deep-water rice

Anabaena sp. grew with mono- and di-ester phosphate compounds as sources of phosphate, indicating the presence of phosphomonoesterase (PMEase) and phosphodiesterase (PDEase) activities. Cell-bound PMEase and PDEase activities were detected during growth in 0.5 and 10 mg PO₄l^–1 only when the cellular phosphate concentration fell to 0.46% of cell protein and the activities increased as cellular phosphate content decreased. The K_m values for these enzymes were 0.3mm forp-nitrophenyl phosphate and 0.2mm for bis-p-nitrophenyl phosphate, respectively. Only PMEase activity was found extracellularly. The pH optima for PMEase and PDEase were 10.2 and 10.4, respectively, and the temperature optima at pH 10.2 were 37°C and 40°C, respectively. Ca²⁺ increased the enzyme activities while Zn²⁺ caused marked inhibition. The inorganic phosphate repressed the cellular PMEase activity after a lag of 4 h.     

Modification of the 1979 "Denver wire code" for different wire or plumbing types.

This article examines "wire configuration coding" as used to estimate relative residential AC magnetic field exposure in epidemiological studies--and the need to alter such coding for time or locations other than those in which the code was developed. Effects of different secondary wire practices are particularly examined.     

Anatomy,neurophysiology and functional aspects of the nucleus isthmi in salamanders of the family Plethodontidae

Summary Tongue-projecting plethodontid salamanders have massive direct ipsilateral retinal afferents to the tectum opticum as well as a large and well developed nucleus isthmi. Retrograde staining revealed two subnuclei: A ventral one projecting to the contralateral tectal hemisphere and a dorsal one projecting back to the ipsilateral side. The isthmic nuclei show a retinotopic organization, which is in register with that of the tectum. Electrophysiological recordings from nucleus-isthmi neurons revealed response properties that are very similar to those found in tectal neurons. Thus, there is no substantial processing of tectal neural activity in the nucleus isthmi. Measurements of peak latencies after electrical and light stimulation suggest the continuous coexistence of 4 representations of the visual field in the tectum mediated by (1) the contralateral and (2) the ipsilateral direct retinal afferents, (3) the uncrossed and (4) the crossed isthmo-tectal projection. (1) and (2) originate at the same moment in the retina and arrive simultaneously in the tectum. It is assumed that in plethodontid salamanders with massive ipsilateral retino-tectal projections depth perception based on disparity cues is achieved by comparison of these images.Representations mediated by (3) and (4) arriving in the tectum at the same time as (1) and (2) originate 10–30 ms earlier in the retina. It is hypothesized that these time differences between (1)/(2) and (3)/(4) are used to calculate three-dimensional trajectories of fast-moving prey objects.Abbreviations EL edge length - FDA fluoresceine dextranamine - RDA tetramethylrhodamine dextranamine - RF receptive field     

Further studies of human whole-body radiofrequency absorption rates

Further studies of human whole-body radiofrequency (RF) absorption rates were carried out using a TEM-cell exposure system. Experiments were done at one frequency near the grounded resonance frequency (approximately 40 MHz), and at several below-resonance frequencies. Absorption rates are small for the K and H orientations of the body, even when grounded. For the body trunk in an E orientation, the absorption rate of a sitting person is about half of the rate for the same person standing with arms at the sides; the latter in turn is about half the rate for the same subject standing with arms over the head. Two-body interactions cause no increase in absorption rates for grounded people. They do, however, increase the absorption rates for subjects in an E orientation in free space; the largest interaction occurs when one subject is lambda/2 behind the other (as seen by the incident wave). When these results are applied to practical occupational exposure situations, the whole-body specific absorption rate does not exceed the ANSI limit of 0.4 W/kg for exposures permitted by the ANSI standard (C95.1-1982) at frequencies from 7 to 40 MHz.     

60-Hz electric fields: detection by female rats

Female rats were trained to detect a vertical, 60-Hz electric field using the same apparatus and procedure we used previously to study behavioral detection of the field by male rats. Each rat was trained individually to press a lever in the presence of the field and not to press in its absence. Correct detections occasionally produced a food pellet. The probability of detecting the field increased as field strength increased. The threshold of detection--ie, the field strength required for detections at a probability of 0.5 after correction for errors--varied among rats between 3 and 10 kV/m. Behavioral detection by female rats was indistinguishable from that by male rats.     

Biological effects of high-frequency electromagnetic fields on Salmonella typhimurium and Drosophila melanogaster

Salmonella typhimurium and Drosophila melanogaster were exposed to continuous wave (CW) 2.45-GHz electromagnetic radiation, pulsed 3.10-GHz electromagnetic radiation, CW 27.12-MHz magnetic fields, or CW 27.12-MHz electric fields (only Drosophila). The temperatures of the treated sample and the nonexposed control sample were kept constant. The temperature difference between exposed and control samples was less than +/- 0.3 degrees C. Ames' assays were made on bacteria that had been exposed to microwaves (SAR 60-130 W/kg) or RF fields (SAR up to 20 W/kg) when growing exponentially in nutrient broth. Survival and number of induced revertants to histidine prototrophy were determined by common plating techniques on rich and minimal agar plates. The Drosophila test consisted of a sensitive somatic system where the mutagenicity was measured by means of mutations in a gene-controlling eye pigmentation. In none of these test systems did microwave or radiofrequency fields induce an elevated mutation frequency. However, a significantly higher concentration of cells was found in the bacterial cultures exposed to the 27-MHz magnetic field or 2.45-GHz CW and 3.10-GHz pulsed microwave radiation.