Distant Nuclei Research Articles

Neuronal expression of herpes simplex virus-1 VP16 protein induces pseudorabies virus escape from silencing and reactivation.

Alpha herpesvirus (α-HV) particles enter their hosts from mucosal surfaces and efficiently maintain fast transport in peripheral nervous system (PNS) axons to establish infections in the peripheral ganglia. The path from axons to distant neuronal nuclei is challenging to dissect due to the difficulty of monitoring early events in a dispersed neuron culture model. We have established well-controlled, reproducible, and reactivateable latent infections in compartmented rodent neurons by infecting physically isolated axons with a small number of viral particles. This system not only recapitulates the physiological infection route but also facilitates independent treatment of isolated cell bodies or axons. Consequently, this system enables study not only of the stimuli that promote reactivation but also the factors that regulate the initial switch from productive to latent infection. Adeno-associated virus (AAV)-mediated expression of herpes simplex-1 (HSV-1) VP16 alone in neuronal cell bodies enabled the escape from silencing of incoming pseudorabies virus (PRV) genomes. Furthermore, the expression of HSV VP16 alone reactivated a latent PRV infection in this system. Surprisingly, the expression of PRV VP16 protein supported neither PRV escape from silencing nor reactivation. We compared transcription transactivation activity of both VP16 proteins in primary neurons by RNA sequencing and found that these homolog viral proteins produce different gene expression profiles. AAV-transduced HSV VP16 specifically induced the expression of proto-oncogenes including c-Jun and Pim2. In addition, HSV VP16 induces phosphorylation of c-Jun in neurons, and when this activity is inhibited, escape of PRV silencing is dramatically reduced.IMPORTANCEDuring latency, alpha herpesvirus genomes are silenced yet retain the capacity to reactivate. Currently, host and viral protein interactions that determine the establishment of latency, induce escape from genome silencing or reactivation are not completely understood. By using a compartmented neuronal culture model of latency, we investigated the effect of the viral transcriptional activator, VP16 on pseudorabies virus (PRV) escape from genome silencing. This model recapitulates the physiological infection route and enables the study of the stimuli that regulate the initial switch from a latent to productive infection. We investigated the neuronal transcriptional activation profiles of two homolog VP16 proteins (encoded by HSV-1 or PRV) and found distinct gene activation signatures leading to diverse infection outcomes. This study contributes to understanding of how alpha herpesvirus proteins modulate neuronal gene expression leading to the initiation of a productive or a latent infection.

Impact of Zeeman and hyperfine interactions on the magnetic properties of paramagnetic metal ions: II. Super-hyperfine interactions with surrounding nuclei

The anisotropic Zeeman and strong hyperfine interactions of a Kramers ion can significantly affect its magnetic properties as well as the interactions with the nearby nuclei. The interactions with the local environment are described in the preceding article. In the current work, the change of the spin states of distant nuclei is studied. Analytical expressions describing the depth of the electron spin echo envelope modulation (ESEEM) are obtained for the ions with anisotropic Zeeman and strong hyperfine interactions. Due to the g-tensor anisotropy, the electron Zeeman interaction axis is tilted in respect to the direction of the external magnetic field which makes non-collinear with the Zeeman interaction axes of the ion and the nearby nuclei and significantly modifies the nuclear spin states. Thus, the isotropic hyperfine interaction, and particularly the Fermi contact interaction can directly contribute to the ESEEM. An additional factor, that can significantly modify the ESEEM signal is the mixing of the multiple oscillations arising when several nuclei occur in optimal conditions for the generation of the nuclear coherence. This situation arises when several EPR transitions of the ion covering a wide range of magnetic fields are examined.

The first mitotic division: a perilous bridge connecting the zygote and the early embryo.

Human embryos are very frequently affected by maternally inherited aneuploidies, which in the vast majority of cases determine developmental failure at pre- or post-implantation stages. However, recent evidence, generated by the alliance between diverse technologies now routinely employed in the IVF laboratory, has revealed a broader, more complex scenario. Aberrant patterns occurring at the cellular or molecular level can impact at multiple stages of the trajectory of development to blastocyst. In this context, fertilization is an extremely delicate phase, as it marks the transition between gametic and embryonic life. Centrosomes, essential for mitosis, are assembled ex novo from components of both parents. Very large and initially distant nuclei (the pronuclei) are brought together and positioned centrally. The overall cell arrangement is converted from being asymmetric to symmetric. The maternal and paternal chromosome sets, initially separate and scattered within their respective pronuclei, become clustered where the pronuclei juxtapose, to facilitate their assembly in the mitotic spindle. The meiotic spindle is replaced by a segregation machinery that may form as a transient or persistent dual mitotic spindle. Maternal proteins assist the decay of maternal mRNAs to allow the translation of newly synthesized zygotic transcripts. The diversity and complexity of these events, regulated in a precise temporal order and occurring in narrow time windows, make fertilization a highly error-prone process. As a consequence, at the first mitotic division, cellular or genomic integrity may be lost, with fatal consequences for embryonic development.

Hyperfine couplings as a probe of orbital anisotropy in heavy-fermion materials

The transferred hyperfine interaction between nuclear and electron spins in an heavy fermion material depends on the hybridization between the $f$-electron orbitals and those surrounding a distant nucleus. In CeMIn$_5$ (M=Rh, Ir, Co), both the hyperfine coupling to the two indium sites as well as the crystalline electric field at the Ce are strongly dependent on the transition metal. We measure a series of CeRh$_{1-x}$Ir$_x$In$_5$ crystals and find that the hyperfine coupling reflects the orbital anisotropy of the ground state Ce 4$f$ wavefunction. These findings provide direct proof that the localized to itinerant transition is dominated by hybridization out of the Ce-In plane in this system.

Submillimeter imaging of the Galactic Center starburst Sgr B2

Context. Star-forming galaxies emit bright molecular and atomic lines in the submillimeter and far-infrared (FIR) domains. However, it is not always clear which gas heating mechanisms dominate and which feedback processes drive their excitation. Aims. The Sgr B2 complex is an excellent template to spatially resolve the main OB-type star-forming cores from the extended cloud environment and to study the properties of the warm molecular gas in conditions likely prevailing in distant extragalactic nuclei. Methods. We present 168 arcmin2 spectral images of Sgr B2 taken with Herschel/SPIRE-FTS in the complete ~450−1545 GHz band. We detect ubiquitous emission from mid-J CO (up to J = 12−11), H2O 21,1−20,2, [C I] 492, 809 GHz, and [N II] 205 μm lines. We also present velocity-resolved maps of the SiO (2−1), N2H+, HCN, and HCO+ (1−0) emission obtained with the IRAM 30 m telescope. Results. The cloud environment (~1000 pc2 around the main cores) dominates the emitted FIR (~80%), H2O 752 GHz (~60%) mid-J CO (~91%), [C I] (~93%), and [N II] 205 μm (~95%) luminosity. The region shows very extended [N II] 205 μm emission (spatially correlated with the 24 and 70 μm dust emission) that traces an extended component of diffuse ionized gas of low ionization parameter (U ≃ 10−3) and low LFIR / MH2 ≃ 4−11 L⊙M⊙−1 ratios (scaling as ∝Tdust6). The observed FIR luminosities imply a flux of nonionizing photons equivalent to G0 ≈ 103. All these diagnostics suggest that the complex is clumpy and this allows UV photons from young massive stars to escape from their natal molecular cores. The extended [C I] emission arises from a pervasive component of neutral gas with nH ≃ 103 cm−3. The high ionization rates in the region, produced by enhanced cosmic-ray (CR) fluxes, drive the gas heating in this component to Tk ≃ 40−60 K. The mid-J CO emission arises from a similarly extended but more pressurized gas component (Pth / k ≃ 107 K cm−3): spatially unresolved clumps, thin sheets, or filaments of UV-illuminated compressed gas (nH ≃ 106 cm−3). Specific regions of enhanced SiO emission and high CO-to-FIR intensity ratios (ICO / IFIR ≳ 10−3) show mid-J CO emission compatible with C-type shock models. A major difference compared to more quiescent star-forming clouds in the disk of our Galaxy is the extended nature of the SiO and N2H+ emission in Sgr B2. This can be explained by the presence of cloud-scale shocks, induced by cloud-cloud collisions and stellar feedback, and the much higher CR ionization rate (>10−15 s−1) leading to overabundant H3+ and N2H+. Conclusions. Sgr B2 hosts a more extreme environment than star-forming regions in the disk of the Galaxy. As a usual template for extragalactic comparisons, Sgr B2 shows more similarities to nearby ultra luminous infrared galaxies such as Arp 220, including a “deficit” in the [C I] / FIR and [N II] / FIR intensity ratios, than to pure starburst galaxies such as M 82. However, it is the extended cloud environment, rather than the cores, that serves as a useful template when telescopes do not resolve such extended regions in galaxies.

Territory occupation and transportation sustainability in Madrid

The usage of private cars contributes significantly to GHG emissions. This study examines the impact of changes in the land occupancy model in the Community of Madrid over the past two decades on mobility. It has resulted in a high consumption of land in the past two decades, but in a more peripheral manner, in increasingly distant nuclei, which may be explained in part by the rise in single-family dwelling. These functionally specialized growths have been sustained by a network of radial highways, resulting in a loss of density, variety, and the compact Mediterranean city model. Simultaneously, it is confirmed that the relocation of the industry and the set of increasingly peripheral and unconnected employment to the residential space, together with the change in the labor model and the scarce residential mobility, the product of a housing in property model, generates an increase in mobility due to work. Due to the difficulty of providing assistance with public transportation, the development of retail malls that are more peripheral, dependent on the road network, and unconnected to the home, leads to an increase in private vehicle travel. These developments have resulted in an increase in private car travel, with a clear loss of the proportion of public transportation, both of which contribute substantially to the production of GHGs and therefore to climate change, as well as excessive energy consumption. As a result, the land occupancy model has a noticeable impact on metropolitan mobility, necessitating cooperation between the urban and transportation models in order to achieve a more balanced region.

Genetic Reporters of Neuronal Activity: c-Fos and G-CaMP6.

The majority of 20th century investigations into anesthetic effects on the nervous system have used electrophysiology. Yet some fundamental limitations to electrophysiologic recordings, including the invasiveness of the technique, the need to place (potentially several) electrodes in every site of interest, and the difficulty of selectively recording from individual cell types, have driven the development of alternative methods for detecting neuronal activation. Two such alternative methods with cellular scale resolution have matured in the last few decades and will be reviewed here: the transcription of immediate early genes, foremost c-fos, and the influx of calcium into neurons as reported by genetically encoded calcium indicators, foremost GCaMP6. Reporters of c-fos allow detection of transcriptional activation even in deep or distant nuclei, without requiring the accurate targeting of multiple electrodes at long distances. The temporal resolution of c-fos is limited due to its dependence upon the detection of transcriptional activation through immunohistochemical assays, though the development of RT-PCR probes has shifted the temporal resolution of the assay when tissues of interest can be isolated. GCaMP6 has several isoforms that trade-off temporal resolution for signal to noise, but the fastest are capable of resolving individual action potential events, provided the microscope used scans quickly enough. GCaMP6 expression can be selectively targeted to neuronal populations of interest, and potentially thousands of neurons can be captured within a single frame, allowing the neuron-by-neuron reporting of circuit dynamics on a scale that is difficult to capture with electrophysiology, as long as the populations are optically accessible.

Highligths from the Fermi Large Area Telescope

The Fermi mission is operating in low Earth orbit since June 2008. Thanks to its large acceptance, the Large Area Telescope (LAT), the on-board pair-conversion telescope for high-energy electromagnetic radiation, has collected more than one billion photons from the whole sky above 20 MeV. The LAT also provided the largest high-energy cosmic-ray electron sample to date, with about 10k events above 1 TeV.This unique database allows the study of thousands of gamma-ray sources of very different nature, from our own Galaxy to distant and active galactic nuclei, as well as addressing fundamental questions of particle astrophysics like the nature of dark matter and the origin of energetic gamma-ray bursts. In addition, cosmic-ray electrons provide a unique probe of the origin and propagation mechanisms of cosmic rays in local galactic environment.

Mechanism of Traumatic Brain Injury at Distant Locations After Exposure to Blast Waves: Preliminary Results from Animal and Phantom Experiments.

Purpose Primary blast-induced traumatic brain injury (bTBI) is the least understood of the four phases of blast injury. Distant injury induced by the blast wave, on the opposite side from the wave entry, is not well understood. This study investigated the mechanism of distant injury in bTBI. Materials and Methods Eight 8-week-old male Sprague-Dawley rats were divided into two groups: group 1 served as the control group and did not receive any shock wave (SW) exposure; group 2 was exposed to SWs (12.5 ± 2.5 MPa). Propagation of SWs within a brain phantom was evaluated by visualization, pressure measurement, and numerical simulation. Results Intracerebral hemorrhage near the ignition site and elongation of the distant nucleus were observed, despite no apparent damage between the two locations in the animal experiment. Visualization, pressure measurement, and numerical simulation indicated the presence of complex wave dynamics accompanying a sudden increase in pressure, followed by negative pressure in the phantom experiment. Conclusion A local increase in pressure above the threshold caused by interference of reflection and rarefaction waves in the vicinity of the brain-skull surface may cause distant injury in bTBI.

Pulse Electron Double Resonance Detected Multinuclear NMR Spectra of Distant and Low Sensitivity Nuclei and Its Application to the Structure of Mn(II) Centers in Organisms.

The ability to characterize the structure of metal centers beyond their primary ligands is important to understanding their chemistry. High-magnetic-field pulsed electron double resonance detected NMR (ELDOR-NMR) is shown to be a very sensitive approach to measuring the multinuclear NMR spectra of the nuclei surrounding Mn(II) ions. Resolved spectra of intact organisms with resonances arising from (55)Mn, (31)P, (1)H, (39)K, (35)Cl, (23)Na, and (14)N nuclei surrounding Mn(2+) centers were obtained. Naturally abundant cellular (13)C could be routinely measured as well. The amplitudes of the (14)N and (2)H ELDOR-NMR spectra were found to be linearly dependent on the number of nuclei in the ligand sphere. The evolution of the Mn(II) ELDOR-NMR spectra as a function of excitation time was found to be best described by a saturation phenomenon rather than a coherently driven process. Mn(II) ELDOR-NMR revealed details about not only the immediate ligands to the Mn(II) ions but also more distant nuclei, providing a view of their extended structures. This will be important for understanding the speciation and chemistry of the manganese complexes as well as other metals found in organisms.

The first estimate of radio jet proper motion at z > 5

Abstract The extremely high-redshift (z = 5.3) radio source SDSS J102623.61+254259.5 (J1026+2542) is among the most distant and most luminous radio-loud active galactic nuclei (AGN) known to date. Its one-sided radio jet structure on milliarcsecond (mas) and ∼10-mas scales typical for blazars was first imaged at 5 GHz with very long baseline interferometry (VLBI) in 2006. Here we report on our dual-frequency (1.7 and 5 GHz) imaging observations performed with the European VLBI Network (EVN) in 2013. The prominent jet structure allows us to identify individual components whose apparent displacement can be detected over the time span of 7.33 yr. This is the first time when jet proper motions are directly derived in a blazar at z > 5. The small values of up to ∼0.1 mas yr−1 are consistent with what is expected in a relativistic cosmological model if redshift is a measure of distance. The apparent superluminal jet speeds, considered tentative because derived from two epochs only, exceed 10 c for three different components along the jet. Based on modelling its spectral energy distribution, J1026+2542 is known to have its X-ray jet oriented close to the line of sight, with significant Doppler boosting and a large bulk Lorentz factor (Γ ≈ 13). The new VLBI observations, indicating ∼2.3 × 1012 K lower limit to the core brightness temperature, are consistent with this picture. The spectral index in the core region is −0.35.

Forest species colonizing cerrado open areas: distance and area effects on the nucleation process

Nucleation is a natural process in which trees scattered in the landscape promote changes in the surrounding environment, which increases the probability of seedling and sapling colonization in a nucleus area. This study aimed to describe forest species community under the crown of Bowdichia virgilioides (Fabaceae) in cerrado sensu stricto, testing two hypotheses: (a) distance: more distant nuclei will have fewer numbers of colonizing species and individuals; (b) crown area: nuclei with larger crown will have more individuals and species. We selected six B. virgilioides nuclei at Estacao Ecologica do Panga (EEP) in which we traced two transects (D1 and D2) and calculated crown area by ellipse formula. In the under-crown area, we identified all individuals (regardless of size) of tree forest species and also measured their height and diameter at stem base. We performed a Pearson correlation between crown area and the number of individuals and species, and between distance from nucleus to closest forest edge and density. In the six nuclei, we found 517 individuals of 24 species, distributed in 14 families, with a total density of 2.48 individuals per square meter. Correlation between crown area and the number of individuals was statistically significant (r = 0.80; n = 6; p = 0.05). There was no correlation statistically significant between crown area and number of species (r = 0.29; n = 6; p > 0.05). There was a negative correlation between density of individuals and the distance from nucleus to forest edge (r = −0.79; n = 6; p = 0.05). This study shows that some forest species were capable to colonize the area under the crown of B. vigilioides in cerrado, facilitated by this nucletor specie. This colonization process was influenced by the environmental changes promoted by B virgilioides, nuclei crown size, distance of nuclei from propagule source, and the capability of some forest species to survive in an environment different from forest.

Proton-driven spin diffusion in rotating solids via reversible and irreversible quantum dynamics

Proton-driven spin diffusion (PDSD) experiments in rotating solids have received a great deal of attention as a potential source of distance constraints in large biomolecules. However, the quantitative relationship between the molecular structure and observed spin diffusion has remained obscure due to the lack of an accurate theoretical description of the spin dynamics in these experiments. We start with presenting a detailed relaxation theory of PDSD in rotating solids that provides such a description. The theory applies to both conventional and radio-frequency-assisted PDSD experiments and extends to the non-Markovian regime to include such phenomena as rotational resonance (R(2)). The basic kinetic equation of the theory in the non-Markovian regime has the form of a memory function equation, with the role of the memory function played by the correlation function. The key assumption used in the derivation of this equation expresses the intuitive notion of the irreversible dissipation of coherences in macroscopic systems. Accurate expressions for the correlation functions and for the spin diffusion constants are given. The theory predicts that the spin diffusion constants governing the multi-site PDSD can be approximated by the constants observed in the two-site diffusion. Direct numerical simulations of PDSD dynamics via reversible Liouville-von Neumann equation are presented to support and compliment the theory. Remarkably, an exponential decay of the difference magnetization can be observed in such simulations in systems consisting of only 12 spins. This is a unique example of a real physical system whose typically macroscopic and apparently irreversible behavior can be traced via reversible microscopic dynamics. An accurate value for the spin diffusion constant can be usually obtained through direct simulations of PDSD in systems consisting of two (13)C nuclei and about ten (1)H nuclei from their nearest environment. Spin diffusion constants computed by this method are in excellent agreement with the spin diffusion constants obtained through equations given by the relaxation theory of PDSD. The constants resulting from these two approaches were also in excellent agreement with the results of 2D rotary resonance recoupling proton-driven spin diffusion (R(3)-PDSD) experiments performed in three model compounds, where magnetization exchange occurred over distances up to 4.9 Å. With the methodology presented, highly accurate internuclear distances can be extracted from such data. Relayed transfer of magnetization between distant nuclei appears to be the main (and apparently resolvable) source of uncertainty in such measurements. The non-Markovian kinetic equation was applied to the analysis of the R(2) spin dynamics. The conventional semi-phenomenological treatment of relxation in R(2) has been shown to be equivalent to the assumption of the Lorentzian spectral density function in the relaxatoin theory of PDSD. As this assumption is a poor approximation in real physical systems, the conventional R(2) treatment is likely to carry a significant model error that has not been recognized previously. The relaxation theory of PDSD appears to provide an accurate, parameter-free alternative. Predictions of this theory agreed well with the full quantum mechanical simulations of the R(2) dynamics in the few simple model systems we considered.

Chemical shift prediction for protein structure calculation and quality assessment using an optimally parameterized force field

The exquisite sensitivity of chemical shifts as reporters of structural information, and the ability to measure them routinely and accurately, gives great import to formulations that elucidate the structure-chemical-shift relationship. Here we present a new and highly accurate, precise, and robust formulation for the prediction of NMR chemical shifts from protein structures. Our approach, shAIC (shift prediction guided by Akaikes Information Criterion), capitalizes on mathematical ideas and an information-theoretic principle, to represent the functional form of the relationship between structure and chemical shift as a parsimonious sum of smooth analytical potentials which optimally takes into account short-, medium-, and long-range parameters in a nuclei-specific manner to capture potential chemical shift perturbations caused by distant nuclei. shAIC outperforms the state-of-the-art methods that use analytical formulations. Moreover, for structures derived by NMR or structures with novel folds, shAIC delivers better overall results; even when it is compared to sophisticated machine learning approaches. shAIC provides for a computationally lightweight implementation that is unimpeded by molecular size, making it an ideal for use as a force field.

Abstract B38: Validation of a nuclear morphometric signature for prostate cancer risk in benign biopsies

Abstract Purpose: Subtle changes in nuclear shape, size and texture precede the histological recognition of prostate cancer (PCa) and thus might provide a useful biomarker for high-risk benign tissue. The objective of this study was to develop and validate a multifeature nuclear score based on direct DNA staining and test its association with field effects and the subsequent detection of PCa in benign biopsies. Methods: Whole tissue sections from 39 radical prostatectomy subjects were Feulgen-stained and scanned (400x) on a digital microscope, allowing maps of DNA content per nuclear pixel to be generated in Matlab@. Samples of PCa and benign epithelial nuclei were randomly selected for measurement of 51 morphometric features. Logistic regression models for discriminating benign from PCa nuclei were built and cross-validated by AUC analysis. Subject-level models, for discriminating benign from malignant nuclear populations taken from individual subjects, were also built and similarly validated. Both backwards elimination and all-subsets approaches were used to select covariates. Nuclear populations were randomly collected < 1mm or > 5mm from cancer foci, and from cancer-free prostates, HGPIN, and PCa Gleason grade 3-5. In addition, nuclear images were collected from negative biopsy cases and controls matched on age and date of biopsy (20 pairs); cases had PCa detected at least 2 years later, controls remained cancer-free and had at least 2 additional negative biopsies. For the case-control study, nuclei were selected using an automatic process that correlated well with results from manual selection. Frequency distributions for multifeature nuclear score from various tissue compartments were compared using the Kolmogorov D statistic. Scores for cases vs. controls were compared using a t test for paired data. Results: In prostatectomy samples, both nuclear- and populationlevel models revealed cancer-like features in benign nuclei adjacent to PCa, compared to nuclei that were more distant or were obtained from PCa-free glands. The score distribution for distant nuclei was also significantly shifted compared to nuclei from cancer-free glands. Interestingly, the best model by leave-one-out cross-validation (AUC = 0.91, 95% CI: 0.81-1.00) included only 5 variance features, reflecting the importance of nuclear pleomorphism. In negative biopsies, a validated model with 5 variance features (including features related to DNA content, size and texture) yielded significantly higher scores in cases than controls (P=0.026). This model had an AUC = 0.71 (95% CI: 0.54-0.87) for discriminating cases from controls. Conclusions: A multifeature nuclear morphometric score, obtained by automated digital analysis, was validated for discrimination of benign from cancer nuclei. This score demonstrated field effects in benign epithelial nuclei at varying distance from PCa lesions, and was associated with subsequent PCa detection in negative biopsies. This biomarker shows definite promise as an intermediate endpoint in chemoprevention trials; however, its utility as an independent predictor of PCa in the clinical setting requires much further study. Citation Information: Cancer Prev Res 2010;3(12 Suppl):B38.

ENDOR Spectroscopy Shows That Guanine N1 Binds to [4Fe−4S] Cluster II of theS-Adenosylmethionine-Dependent Enzyme MoaA: Mechanistic Implications

The S-adenosylmethionine-dependent enzyme MoaA, in concert with MoaC, catalyzes the first step of molybdenum cofactor biosynthesis, the conversion of guanosine 5'-triphosphate (5'-GTP) into precursor Z. A published X-ray crystal structure of MoaA with the substrate 5'-GTP revealed that the substrate might be bound to the unique iron of one of two 4Fe-4S clusters through either or both the amino and N1 nitrogen nuclei. Use of 35 GHz continuous-wave ENDOR spectroscopy of MoaA with unlabeled and (15)N-labeled substrate and a reduced [4Fe-4S](+) cluster now demonstrates that only one nitrogen nucleus is bound to the cluster. Experiments with the substrate analogue inosine 5'-triphosphate further demonstrate that it is the N1 nitrogen that binds. Two of the more distant nitrogen nuclei have also been detected by 35 GHz pulsed ENDOR spectroscopy, allowing a rough approximation of their distances from the cluster to be calculated. Combining this information with the crystal structure, we propose that the guanine base adopts the enol tautomer as N1 binds to Fe4 and the O6-H hydroxyl group forms a hydrogen bond with S4 of the 4Fe-4S cluster, and that this binding-induced tautomerization may have important mechanistic ramifications.

ESEEM Measurements of Local Water Concentration in D2O-Containing Spin-Labeled Systems

To calibrate electron spin echo envelope modulation (ESEEM) amplitudes with respect to the deuterium water content in spin-labeled biological systems, ESEEM of nitroxide TEMPO has been studied in frozen glassy D2O-dimethylsulfoxide mixtures of different composition. The interaction between the unpaired electron of nitroxide and the deuterium nuclei manifests itself in a cosine Fourier transform spectrum as the sum of a narrow line with the doublet quadrupole splitting and of a broad one. The narrow line arises from interaction with distant deuterium nuclei, the broad one arises from interaction with nearby nuclei belonging to nitroxide-water molecule complexes. The dependence on water concentration was found to be nonlinear for the intensity of the narrow line and close to linear for the intensity of the quadrupole doublet. Therefore, the intensity of the quadrupole doublet is suggested as a measure of concentration of free water around a spin label in biological objects. Fourier transform line shape was theoretically simulated for different model distributions of water molecules around the spin label. Simulations confirm the linear dependence of the quadrupole doublet intensity on water concentration seen in the experiment. The suggested approach was applied to analyze data for spin-labeled dipalmitoylphosphatidylcholine (DPPC) and DPPC-cholesterol D2O-hydrated model membranes. The concentration of free water near the spin-labeled fourth carbon atom along the lipid chain was estimated as 5.2 and 7.2 M for DPPC and DPPC-cholesterol membranes, respectively.

Localization of CD226 in the mouse hippocampus and cerebellum during adulthood and postnatal development

CD226, a member of cell adhesion molecules, has been widely studied in the immune system; however, its expression in the CNS remains unknown. In our present study, we detected CD226 mRNA and protein in the mouse hippocampus and cerebellum by RT-PCR and Western blotting, respectively. Immunohistochemical studies found that CD226 is primarily located in the hilus of the dentate gyrus and stratum lucidum aligned along the pyramidal cells in the hippocampal CA3 area, the interspaces of granular cells and the somata of the Purkinje cells in the cerebellar cortex during adulthood. Double-staining results revealed that CD226 co-localized well with synaptic marker proteins including synaptophysin, syntaxin and PSD-95. During postnatal development, CD226 could not be detected at its adult locations until postnatal day 12; however, it was temporally expressed in the somata of neighboring or distant nuclei associated with its adult location. These results showed the diverse localization of CD226 in the mouse hippocampus and cerebellum for the first time and suggested its potential role in the CNS.

Accurate and efficient treatment of two-electron contributions in quasirelativistic high-order Douglas-Kroll density-functional calculations

Two-component quasirelativistic approaches are in principle capable of reproducing results from fully relativistic calculations based on the four-component Dirac equation (with fixed particle number). For one-electron systems, this also holds in practice, but in many-electron systems one has to transform the two-electron interaction, which is necessary because a picture change occurs when going from the Dirac equation to a two-component method. For one-electron properties, one can take full account of picture change in a manageable way, but for the electron interaction, this would spoil the computational advantages which are the main reason to perform quasirelativistic calculations. Exploiting those picture change effects are largest in the atomic cores, which in molecular applications do not differ too much from the cores of isolated neutral atoms, we propose an elegant, efficient, and accurate approximation to the two-electron picture change problem. The new approach, called the "model potential" approach because it makes use of atomic (four- and two-component) data to estimate picture change effects in molecules, shares with the nuclear-only approach that the Douglas-Kroll operator needs to be constructed only once (not in each self-consistent-field iteration) and that no time-consuming multicenter relativistic two-electron integrals need to be calculated. The new approach correctly describes the screening of both the nearest nucleus and distant nuclei, for the scalar-relativistic as well as the spin-orbit parts of the Hamiltonian. The approach is tested on atomic and molecular-orbital energies as well as spectroscopic constants of the lead dimer.

Long range superhyperfine interactions in polycrystalline vanadium doped SnO 2 investigated by CW and pulsed ENDOR spectroscopy

CW and pulsed ENDOR was used to probe the electron nuclear superhyperfine interactions between V 4+ ions and distant Sn nuclei in vanadium doped tin oxide (V/SnO 2). Whilst interactions with two sets of nearest neighbour Sn nuclei (with a V–Sn distance of 3.185 and 3.708 Å respectively) are observed by EPR, superhyperfine couplings to two remote sets of tins (with a V–Sn distance of 6.370 and ∼7.42 Å) are detected by ENDOR. The interaction was found to be largely isotropic and largest along the crystal c axis. Small differences in the remote tin environments were also detected by ENDOR.