X-ray Thomson scattering absolute intensity from the f-sum rule in the imaginary-time domain.

We present a formally exact and simulation-free approach for the normalization of X-ray Thomson scattering (XRTS) spectra based on the f-sum rule of the imaginary-time correlation function (ITCF). Our method works for any degree of collectivity, over a broad range of temperatures, and is applicable even in nonequilibrium situations. In addition to giving us model-free access to electronic correlations, this new approach opens up the intriguing possibility to extract a plethora of physical properties from the ITCF based on XRTS experiments.

Observing the onset of pressure-driven K-shell delocalization.

The gravitational pressure in many astrophysical objects exceeds one gigabar (one billion atmospheres)1-3, creating extreme conditions where the distance between nuclei approaches the size of the K shell. This close proximity modifies these tightly bound states and, above a certain pressure, drives them into a delocalized state4. Both processes substantially affect the equation of state and radiation transport and, therefore, the structure and evolution of these objects. Still, our understanding of this transition is far from satisfactory and experimental data are sparse. Here we report on experiments that create and diagnose matter at pressures exceeding three gigabars at the National Ignition Facility5 where 184 laser beams imploded a beryllium shell. Bright X-ray flashes enable precision radiography and X-ray Thomson scattering that reveal both the macroscopic conditions and the microscopic states. The data show clear signs of quantum-degenerate electrons in states reaching 30 times compression, and a temperature of around two million kelvins. At the most extreme conditions, we observe strongly reduced elastic scattering, which mainly originates from K-shell electrons. We attribute this reduction to the onset of delocalization of the remaining K-shell electron. With this interpretation, the ion charge inferred from the scattering data agrees well with ab initio simulations, but it is significantly higher than widely used analytical models predict6.

Absolute Equation-of-State Measurement for Polystyrene from 25 to 60 Mbar Using a Spherically Converging Shock Wave.

We have developed an experimental platform for the National Ignition Facility that uses spherically converging shock waves for absolute equation-of-state (EOS) measurements along the principal Hugoniot. In this Letter, we present one indirect-drive implosion experiment with a polystyrene sample that employs radiographic compression measurements over a range of shock pressures reaching up to 60 Mbar (6 TPa). This significantly exceeds previously published results obtained on the Nova laser [R. Cauble et al., Phys. Rev. Lett. 80, 1248 (1998)PRLTAO0031-900710.1103/PhysRevLett.80.1248] at a strongly improved precision, allowing us to discriminate between different EOS models. We find excellent agreement with Kohn-Sham density-functional-theory-based molecular dynamics simulations.

Quantum theory for the dynamic structure factor in correlated two-component systems in nonequilibrium: Application to x-ray scattering.

We present a quantum theory for the dynamic structure factors in nonequilibrium, correlated, two-component systems such as plasmas or warm dense matter. The polarization function, which is needed as the input for the calculation of the structure factors, is calculated in nonequilibrium based on a perturbation expansion in the interaction strength. To make our theory applicable for x-ray scattering, a generalized Chihara decomposition for the total electron structure factor in nonequilibrium is derived. Examples are given and the influence of correlations and exchange on the structure and the x-ray-scattering spectrum are discussed for a model nonequilibrium distribution, as often encountered during laser heating of materials, as well as for two-temperature systems.

X-ray scattering measurements on imploding CH spheres at the National Ignition Facility.

We have performed spectrally resolved x-ray scattering measurements on highly compressed polystyrene at pressures of several tens of TPa (100 Mbar) created by spherically convergent shocks at the National Ignition Facility. Scattering data of line radiation at 9.0 keV were recorded from the dense plasma shortly after shock coalescence. Accounting for spatial gradients, opacity effects, and source broadening, we demonstrate the sensitivity of the elastic scattering component to carbon K-shell ionization while at the same time constraining the temperature of the dense plasma. For six times compressed polystyrene, we find an average temperature of 86 eV and carbon ionization state of 4.9, indicating that widely used ionization models need revision in order to be suitable for the extreme states of matter tested in our experiment.

Observation of finite-wavelength screening in high-energy-density matter.

A key component for the description of charged particle systems is the screening of the Coulomb interaction between charge carriers. First investigated in the 1920s by Debye and Hückel for electrolytes, charge screening is important for determining the structural and transport properties of matter as diverse as astrophysical and laboratory plasmas, nuclear matter such as quark-gluon plasmas, electrons in solids, planetary cores and charged macromolecules. For systems with negligible dynamics, screening is still mostly described using a Debye-Hückel-type approach. Here, we report the novel observation of a significant departure from the Debye-Hückel-type model in high-energy-density matter by probing laser-driven, shock-compressed plastic with high-energy X-rays. We use spectrally resolved X-ray scattering in a geometry that enables direct investigation of the screening cloud, and demonstrate that the observed elastic scattering amplitude is only well described within a more general approach.

X-ray continuum emission spectroscopy from hot dense matter at Gbar pressures.

We have measured the time-resolved x-ray continuum emission spectrum of ∼30 times compressed polystyrene created at stagnation of spherically convergent shock waves within the Gbar fundamental science campaign at the National Ignition Facility. From an exponential emission slope between 7.7 keV and 8.1 keV photon energy and using an emission model which accounts for reabsorption, we infer an average electron temperature of 375 ± 21 eV, which is in good agreement with HYDRA-1D simulations.

Observations of continuum depression in warm dense matter with x-ray Thomson scattering.

Detailed measurements of the electron densities, temperatures, and ionization states of compressed CH shells approaching pressures of 50 Mbar are achieved with spectrally resolved x-ray scattering. Laser-produced 9 keV x-rays probe the plasma during the transient state of three-shock coalescence. High signal-to-noise x-ray scattering spectra show direct evidence of continuum depression in highly degenerate warm dense matter states with electron densities ne>1024 cm-3. The measured densities and temperatures agree well with radiation-hydrodynamic modeling when accounting for continuum lowering in calculations that employ detailed configuration accounting.

Reduced coupled-mode approach to electron-ion energy relaxation.

We present a reduced model for the energy transfer via coupled collective modes in two-temperature plasmas based on quantum statistical theory. The model is compared with exact numerical evaluations of the coupled-mode (CM) energy transfer rate and with alternative reduced approaches over a range of conditions in the warm dense matter (WDM) and inertial confinement fusion (ICF) regimes. Our approach shows excellent agreement with an exact treatment of the CM rate and supports the importance of the coupled-mode effect for the temperature and energy relaxation in WDM and ICF plasmas. We find that electronic damping of collective ion density fluctuations is crucial for correctly describing the mode spectrum and, thus, the energy exchange. The reduced CM approach is studied over a wide parameter space, enabling us to establish its limits of applicability.

Analysis of Thomson scattering from nonequilibrium plasmas.

We develop the theory for light scattering as a diagnostic method for plasmas in nonequilibrium states. We show how well-known nonequilibrium features, like beam acoustic modes, arise in the spectra. The analysis of an experiment with strongly driven electrons demonstrates the abilities of the new approach; we find qualitatively different scattering spectra for different times and excellent agreement with the experimental data after time integration. Finally, an analysis of data from dense beryllium suggests that an energetic electron component exists in this experiment as well.

Measurements of electron transport in foils irradiated with a picosecond time scale laser pulse.

The heating of solid foils by a picosecond time scale laser pulse has been studied by using x-ray emission spectroscopy. The target material was plastic foil with a buried layer of a spectroscopic tracer material. The laser pulse length was either 0.5 or 2 ps, which resulted in a laser irradiance that varied over the range 10(16)-10(19) W/cm(2). Time-resolved measurements of the buried layer emission spectra using an ultrafast x-ray streak camera were used to infer the density and temperature conditions as a function of laser parameters and depth of the buried layer. Comparison of the data to different models of electron transport showed that they are consistent with a model of electron transport that predicts the bulk of the target heating is due to return currents.

Pre-treatment of cattle semen or oocytes with purified milk osteopontin affects in vitro fertilization and embryo development.

This study was designed to investigate the effects of pre-incubating cattle spermatozoa or matured oocytes with purified osteopontin (OPN) from cattle milk on fertilization in cattle and embryonic development in vitro. There were two different experiments, semen from six mature Holstein bulls (Bos Taurus) was frozen with different concentrations of OPN (0, 1, 10, 100 microg/mL). Matured cattle oocytes were also pre-treated with OPN (0, 10, 100 microg/mL). In both experiments, pre-treated oocytes or frozen semen, was processed for in vitro fertilization and embryo development. Significantly more oocytes were fertilized when using frozen semen with 10 microg/mL OPN (bull 2=85+/-4% and bull 5=78+/-4%) than without OPN (bull 2=75+/-4% and bull 5=69+/-4%). Those bulls also had increase in cleavage and embryo development (bull 2=85+/-3%, 41+/-1.9%; bull 5=76+/-2%, 37+/-1.8%) compared with control (bull 2=75+/-3%, 30+/-2%; bull 5=68+/-2%, 29+/-2%). Incubating matured oocytes in 10 microg/mL OPN (87+/-3%) and 100 microg/mL OPN (88+/-3%) significantly increased fertilization than control (73+/-3%). OPN also improve cleavage, and embryo development in treatments with 10 microg/mL OPN (82.7+/-1.3%; 31.7+/-1.4%) and 100 microg/mL OPN (85.8+/-1.3%; 33.8+/-1.5%) when compared with control (74.1+/-1.3%; 24.2+/-1.2%). These data suggest that both, spermatozoa from some bulls and oocytes may associate with OPN, suggesting a facilitory role on in vitro fertilization and embryo development.

Induction of the acrosome reaction in bull spermatozoa with plasmin.

Proteolytic enzymes appear to have an essential role in multiple phases of mammalian fertilization. Plasmin, the active enzyme of the plasminogen activation system that stimulates fibrinolysis and proteolysis has a less well-documented role in reproduction. The current study was conducted to investigate the effect of the active protease, plasmin, on the ability of bovine sperm to undergo the acrosome reaction. Aliquots of freshly ejaculated bull sperm were incubated in capacitating conditions with 10 microg ml-1 of heparin for 4 h. Every 2 h an aliquot of spermatozoa was exposed to lysophosphatidylcholine (100 microg ml-1) or 0, 0.1, 1, 10 and 100 mU of plasmin to induce the acrosome reaction in capacitated spermatozoa. Plasmin increased the percentage of live acrosome reacted sperm after 4 h of incubation in the capacitation medium. Viability was not affected by any of the treatments. This study provides new information on bovine acrosome reaction during in vitro incubation with plasmin and indicates that this protease may participate in the proteolytic events that accompany fertilization.

Expression and presence of osteopontin and integrins in the bovine oviduct during the oestrous cycle.

Osteopontin and integrin alpha(v)beta(3) are known to mediate cell-cell attachment and cell migration. Western blot analysis was used to demonstrate the presence of osteopontin in oviductal fluid collected from ampullar and isthmic regions. Three different osteopontin isoforms of 55 kDa, 48 kDa and 25 kDa were detected in the oviductal fluid. Each isoform was observed during the luteal and non-luteal phases and in both ampullar and isthmic fluids. The 25 kDa osteopontin was the most prevalent isoform in oviductal fluid except in isthmic fluid during the non-luteal phase of the oestrous cycle. RT-PCR was performed with RNA from oviductal cells collected from cows in the post-ovulatory, early to mid-luteal, late luteal or pre-ovulatory stages of the oestrous cycle to reveal the oviduct as a site of osteopontin and integrin synthesis. Only one osteopontin mRNA transcript was detected, and amounts did not vary throughout the oestrous cycle. In contrast, the relative expression of the integrin subtypes alpha(v) and beta(1) during the late luteal phase was lower compared with the other oestrous cycle phases. Integrin beta(3) mRNA content increased significantly from the lowest level during the late luteal phase to the highest level before ovulation. In conclusion, differential presence of osteopontin isoforms and integrins in the bovine oviduct throughout the oestrous cycle indicate that osteopontin-integrin interactions have functional roles in normal oviduct physiology which may potentially influence interactions between the gametes, the embryo, and the epithelium.

An extended hydrophobic interactive surface of Yersinia pestis Caf1M chaperone is essential for subunit binding and F1 capsule assembly.

A single polypeptide subunit, Caf1, polymerizes to form a dense, poorly defined structure (F1 capsule) on the surface of Yersinia pestis. The caf-encoded assembly components belong to the chaperone-usher protein family involved in the assembly of composite adhesive pili, but the Caf1M chaperone itself belongs to a distinct subfamily. One unique feature of this subfamily is the possession of a long, variable sequence between the F1 beta-strand and the G1 subunit binding beta-strand (FGL; F1 beta-strand to G1 beta-strand long). Deletion and insertion mutations confirmed that the FGL sequence was not essential for folding of the protein but was absolutely essential for function. Site-specific mutagenesis of individual residues identified Val-126, in particular, together with Val-128 as critical residues for the formation of a stable subunit-chaperone complex and the promotion of surface assembly. Differential effects on periplasmic polymerization of the subunit were also observed with different mutants. Together with the G1 strand, the FGL sequence has the potential to form an interactive surface of five alternating hydrophobic residues on Caf1M chaperone as well as in seven of the 10 other members of the FGL subfamily. Mutation of the absolutely conserved Arg-20 to Ser led to drastic reduction in Caf1 binding and surface assembled polymer. Thus, although Caf1M-Caf1 subunit binding almost certainly involves the basic principle of donor strand complementation elucidated for the PapD-PapK complex, a key feature unique to the chaperones of this subfamily would appear to be capping via high-affinity binding of an extended hydrophobic surface on the respective single subunits.

The identification of early risk factors for severe emotional disturbances and emotional handicaps: an epidemiological approach.

Epidemiological methodology is used to examine the relationship between early childhood risk factors and future identification as having a Severe Emotional Disturbance or as having an Emotional Handicap (SED/EH) at age 13. Data were obtained from 1979/1980 Florida birth records that were electronically linked with 1992/1993 Florida school records. An epidemiological perspective was chosen due to its ability to model both individual and community-level risk. In regards to increasing an individual's risk of SED/EH, two factors, gender (being male) and low maternal education (mother not completing high school at the time of the child's birth), were found to have particularly strong effects. When examining effects of these risk factors upon overall rates of SED/EH in the community, maternal education and marital status (being unmarried at the time of the child's birth) were associated with a large proportion of the cases. Health/biological markers were moderately associated with SED/EH on the individual level, but were related to a relatively small percentage of cases in the population. In addition, effects varied based upon ethnic/cultural heritage. Researchers are encouraged to consider using an epidemiological perspective and its potential utility in the field of community psychology and public policy is discussed.

Structural and functional significance of the FGL sequence of the periplasmic chaperone Caf1M of Yersinia pestis.

The periplasmic molecular chaperone Caf1M of Yersinia pestis is a typical representative of a subfamily of specific chaperones involved in assembly of surface adhesins with a very simple structure. One characteristic feature of this Caf1M-like subfamily is possession of an extended, variable sequence (termed FGL) between the F1 and subunit binding G1 beta-strands. In contrast, FGS subfamily members, characterized by PapD, have a short F1-G1 loop and are involved in assembly of complex pili. To elucidate the structural and functional significance of the FGL sequence, a mutant Caf1M molecule (dCaf1M), in which the 27 amino acid residues between the F1 and G1 beta-strands had been deleted, was constructed. Expression of the mutated caf1M in Escherichia coli resulted in accumulation of high levels of dCaf1M. The far-UV circular dichroism spectra of the mutant and wild-type proteins were indistinguishable and exhibited practically the same temperature and pH dependencies. Thus, the FGL sequence of Caf1M clearly does not contribute significantly to the stability of the protein conformation. Preferential cleavage of Caf1M by trypsin at Lys-119 confirmed surface exposure of this part of the FGL sequence in the isolated chaperone and periplasmic chaperone-subunit complex. There was no evidence of surface-localized Caf1 subunit in the presence of the Caf1A outer membrane protein and dCaf1M. In contrast to Caf1M, dCaf1M was not able to form a stable complex with Caf1 nor could it protect the subunit from proteolytic degradation in vivo. This demonstration that the FGL sequence is required for stable chaperone-subunit interaction, but not for folding of a stable chaperone, provides a sound basis for future detailed molecular analyses of the FGL subfamily of chaperones.

Osteopontin localization in the Holstein bull reproductive tract.

Previously we reported that the 55-kDa fertility-associated protein in Holstein bull seminal plasma (SP) is osteopontin (OPN). The objective of the present study was to localize OPN in tissues and fluids in the Holstein bull reproductive tract to determine its origin. Antisera generated against human recombinant OPN, as well as antiserum prepared against purified bovine seminal plasma OPN, reacted with protein in SP, accessory sex gland fluid, seminal vesicle fluid, ampullary fluid, and urine using one- and two-dimensional SDS-PAGE Western blot analysis. However, these antisera failed to detect OPN in cauda epididymal fluid or solubilized sperm membranes. Immunofluorescence histochemistry localized OPN in the lumen and epithelial cells of the seminal vesicle and ampulla, but not in tissues of testis, epididymis, prostate, and bulbourethral gland. OPN was not detected immunohistochemically in epididymal, ampullary, or ejaculated sperm treated with or without Triton X-100. We concluded that the primary sources of OPN in bull SP are the seminal vesicles and ampulla.

Identification of a fertility-associated protein in bull seminal plasma as lipocalin-type prostaglandin D synthase.

The objective of this study was to characterize a 26-kDa seminal plasma protein previously shown to be prevalent in bulls of high fertility. Spots of this protein, excised and electroeluted from two-dimensional SDS-PAGE gels, were used for N-terminal amino acid sequencing and for preparation of antiserum in rabbits. The N-terminal amino acid sequence (ALQPNFEEDKFLGRWFTSGL) was 75% identical and 100% homologous to lipocalin-type prostaglandin (PG) D synthase isolated from human cerebrospinal fluid (CSF). Western blots of purified 26-kDa protein cross-reacted with polyclonal antibodies against lipocalin-type PGD synthase isolated from rat brain and human CSF. Immunoreactive bands at 26 kDa appeared in Western blots of seminal plasma and cauda epididymal fluid (CEF). A 29-kDa band appeared in blots of rete testis fluid (RTF). PGD synthase activity was detected in seminal plasma, CEF, and RTF. The cDNA for bovine lipocalin-type PGD synthase, isolated by reverse transcription-polymerase chain reaction, contained a coding region of 573 base pairs corresponding to 191 amino acids. The amino acid sequence was 63-80% identical to that of the enzyme of other mammals. These results establish that the 26-kDa fertility-associated protein in bull seminal plasma is lipocalin-type PGD synthase. Although we do not yet know the role of lipocalin-type PGD synthase in the male genital tract, we speculate that this protein may play an important role in both the development and the maturation of sperm.

Influence of the conserved disulphide bond, exposed to the putative binding pocket, on the structure and function of the immunoglobulin-like molecular chaperone Caf1M of Yersinia pestis.

The Yersinia pestis protein Caf1M is a typical representative of a subfamily of periplasmic molecular chaperones with characteristic structural and functional features, one of which is the location of two conserved cysteine residues close to the putative binding pocket. We show that these residues form a disulphide bond, the reduction and alkylation of which significantly increases the dissociation constant of the Caf1M-Caf1 (where Caf 1 is a polypeptide subunit of the capsule) complex [from a Kd of (4.77+/-0.50)x10(-9) M for the intact protein to one of (3.68+/-0.68)x10(-8) M for the modified protein]. The importance of the disulphide bond for the formation of functional Caf1M in vivo was demonstrated using an Escherichia coli dsbA mutant carrying the Y. pestis f1 operon. In accordance with the CD and fluorescence measurements, the disulphide bond is not important for maintenance of the overall structure of the Caf1M molecule, but would appear to affect the fine structural properties of the subunit binding site. A three-dimensional model of the Caf1M-Caf1 complex was designed based on the published crystal structure of PapD (a chaperone required for Pap pili assembly) complexed with a peptide corresponding to the C-terminus of the papG subunit. In the model the disulphide bond is in close proximity to the invariant Caf1M Arg-23 and Lys-142 residues that are assumed to anchor the C-terminal group of the subunit. The importance of this characteristic disulphide bond for the orchestration of the binding site and subunit binding, as well as for the folding of the protein in vivo, is likely to be a common feature of this subfamily of Caf1M-like chaperones. A possible model for the role of the disulphide bond in Caf1 assembly is discussed.