Orbital-specific mapping of the ligand exchange dynamics of Fe(CO)5 in solution.

Transition-metal complexes have long attracted interest for fundamental chemical reactivity studies and possible use in solar energy conversion. Electronic excitation, ligand loss from the metal centre, or a combination of both, creates changes in charge and spin density at the metal site that need to be controlled to optimize complexes for photocatalytic hydrogen production and selective carbon-hydrogen bond activation. An understanding at the molecular level of how transition-metal complexes catalyse reactions, and in particular of the role of the short-lived and reactive intermediate states involved, will be critical for such optimization. However, suitable methods for detailed characterization of electronic excited states have been lacking. Here we show, with the use of X-ray laser-based femtosecond-resolution spectroscopy and advanced quantum chemical theory to probe the reaction dynamics of the benchmark transition-metal complex Fe(CO)5 in solution, that the photo-induced removal of CO generates the 16-electron Fe(CO)4 species, a homogeneous catalyst with an electron deficiency at the Fe centre, in a hitherto unreported excited singlet state that either converts to the triplet ground state or combines with a CO or solvent molecule to regenerate a penta-coordinated Fe species on a sub-picosecond timescale. This finding, which resolves the debate about the relative importance of different spin channels in the photochemistry of Fe(CO)5 (refs 4, 16 - 20), was made possible by the ability of femtosecond X-ray spectroscopy to probe frontier-orbital interactions with atom specificity. We expect the method to be broadly applicable in the chemical sciences, and to complement approaches that probe structural dynamics in ultrafast processes.

Stimulated X-ray emission for materials science.

Resonant inelastic X-ray scattering and X-ray emission spectroscopy can be used to probe the energy and dispersion of the elementary low-energy excitations that govern functionality in matter: vibronic, charge, spin and orbital excitations. A key drawback of resonant inelastic X-ray scattering has been the need for high photon densities to compensate for fluorescence yields of less than a per cent for soft X-rays. Sample damage from the dominant non-radiative decays thus limits the materials to which such techniques can be applied and the spectral resolution that can be obtained. A means of improving the yield is therefore highly desirable. Here we demonstrate stimulated X-ray emission for crystalline silicon at photon densities that are easily achievable with free-electron lasers. The stimulated radiative decay of core excited species at the expense of non-radiative processes reduces sample damage and permits narrow-bandwidth detection in the directed beam of stimulated radiation. We deduce how stimulated X-ray emission can be enhanced by several orders of magnitude to provide, with high yield and reduced sample damage, a superior probe for low-energy excitations and their dispersion in matter. This is the first step to bringing nonlinear X-ray physics in the condensed phase from theory to application.

Time-resolved electron spectroscopy for chemical analysis of photodissociation: Photoelectron spectra of Fe(CO)5, Fe(CO)4, and Fe(CO)3.

The prototypical photoinduced dissociation of Fe(CO)5 in the gas phase is used to test time-resolved x-ray photoelectron spectroscopy for studying photochemical reactions. Upon one-photon excitation at 266 nm, Fe(CO)5 successively dissociates to Fe(CO)4 and Fe(CO)3 along a pathway where both fragments retain the singlet multiplicity of Fe(CO)5. The x-ray free-electron laser FLASH is used to probe the reaction intermediates Fe(CO)4 and Fe(CO)3 with time-resolved valence and core-level photoelectron spectroscopy, and experimental results are interpreted with ab initio quantum chemical calculations. Changes in the valence photoelectron spectra are shown to reflect changes in the valence-orbital interactions upon Fe-CO dissociation, thereby validating fundamental theoretical concepts in Fe-CO bonding. Chemical shifts of CO 3σ inner-valence and Fe 3p core-level binding energies are shown to correlate with changes in the coordination number of the Fe center. We interpret this with coordination-dependent charge localization and core-hole screening based on calculated changes in electron densities upon core-hole creation in the final ionic states. This extends the established capabilities of steady-state electron spectroscopy for chemical analysis to time-resolved investigations. It could also serve as a benchmark for how charge and spin density changes in molecular dissociation and excited-state dynamics are expressed in valence and core-level photoelectron spectroscopy.

Communication: Direct evidence for sequential dissociation of gas-phase Fe(CO)5 via a singlet pathway upon excitation at 266 nm.

We prove the hitherto hypothesized sequential dissociation of Fe(CO)5 in the gas phase upon photoexcitation at 266 nm via a singlet pathway with time-resolved valence and core-level photoelectron spectroscopy with an x-ray free-electron laser. Valence photoelectron spectra are used to identify free CO molecules and to determine the time constants of stepwise dissociation to Fe(CO)4 within the temporal resolution of the experiment and further to Fe(CO)3 within 3 ps. Fe 3p core-level photoelectron spectra directly reflect the singlet spin state of the Fe center in Fe(CO)5, Fe(CO)4, and Fe(CO)3 showing that the dissociation exclusively occurs along a singlet pathway without triplet-state contribution. Our results are important for assessing intra- and intermolecular relaxation processes in the photodissociation dynamics of the prototypical Fe(CO)5 complex in the gas phase and in solution, and they establish time-resolved core-level photoelectron spectroscopy as a powerful tool for determining the multiplicity of transition metals in photochemical reactions of coordination complexes.

Lithium-induced Clock Gene Expression in Lymphoblastoid Cells of Bipolar Affective Patients.

INTRODUCTION: Disturbances of circadian rhythms occur in all episodes of bipolar disorder (BD). Lithium, as gold-standard in the maintenance treatment of BD, is known to influence circadian processes. METHODS: In a pilot study lymphoblastoid cell lines (LCLs) were generated from 8 BD patients and 6 healthy controls. The LCLs were treated with lithiumchloride (LiCl) for 3 weeks. Cell cycles were then synchronized and expressional analysis by quantitative Real Time PCR was done. RESULTS: BD and controls differed in the period length regarding DBP (albumin D-box binding protein) expression and DBP expression was also influenced by lithium treatment. Furthermore, baseline DBP expression was significantly different between non-treated BD and healthy controls. None of the other analyzed circadian genes showed to be influenced by chronic lithium treatment or to be differentially regulated due to the diagnosis. DISCUSSION: We here show that chronic lithium treatment of LCLs leads to decreased expression of the clock gene DBP, rendering DBP a lithium-regulated gene. We could confirm the role of the circadian clock as well in lithium mode of action as in the pathomechanisms of BD although future studies with a greater number of participants and cell lines are needed.

The Early Warning and Response System for communicable diseases in the EU: an overview from 1999 to 2005.

Under Decision 2119/98/EC of the European Parliament and of the Council, a network for epidemiological surveillance and control of communicable diseases in the Community was set up in 1998. One pillar of Decision 2119/98/EC is the early warning and response system (EWRS). The main objective of the network is to establish permanent communication between European Union (EU) Member States' public health authorities, which are responsible for determining the measures required to control communicable disease-related events. Since 1998, a web based informatics tool has been developed in order to allow information to be shared between the relevant public health authorities. Between 1998 and December 2005, a total of 583 messages were circulated through the EWRS, notifying 396 events. The information shared through the system helped to coordinate public health measures in the EU. However, only few events prompted specific measures at Community level and most of them were controlled with public health measures applied at national level. Major events (such as the Severe Acute Respiratory Syndrome) and the results of simulation exercises prompted the Commission to upgrade the informatics system on the basis of user needs. Since 1 May 2004 the 10 newest Member States have provided information under the current legislation and since April 2005 the European Centre for Disease Prevention and Control (ECDC) is part of the system. Future developments will include a link between the existing EWRS and the communication platform currently developed by the ECDC.

The early warning and response system for communicable diseases in the EU: an overview from 1999 to 2005.

Under Decision 2119/98/EC of the European Parliament and of the Council, a network for epidemiological surveillance and control of communicable diseases in the Community was set up in 1998. One pillar of Decision 2119/98/EC is the early warning and response system (EWRS). The main objective of the network is to establish permanent communication between European Union (EU) Member States' public health authorities, which are responsible for determining the measures required to control communicable disease-related events. Since 1998, a web based informatics tool has been developed in order to allow information to be shared between the relevant public health authorities. Between 1998 and December 2005, a total of 583 messages were circulated through the EWRS, notifying 396 events. The information shared through the system helped to coordinate public health measures in the EU. However, only few events prompted specific measures at Community level and most of them were controlled with public health measures applied at national level. Major events (such as the Severe Acute Respiratory Syndrome) and the results of simulation exercises prompted the Commission to upgrade the informatics system on the basis of user needs. Since 1 May 2004 the 10 newest Member States have provided information under the current legislation and since April 2005 the European Centre for Disease Prevention and Control (ECDC) is part of the system. Future developments will include a link between the existing EWRS and the communication platform currently developed by the ECDC.

Salicylic acid inhibits ultraviolet- and cis-platinum-induced human immunodeficiency virus expression.

Previous studies have shown that exposure of HeLa cells stably transfected with an HIV-long terminal repeat-chloramphenicol acetyltransferase (HIV-LTR-CAT) construct to many DNA-damaging agents (such as UV light) induces expression from the HIV LTR. By culturing the cells with salicylic acid we demonstrated dose-dependent repression of this UV-or cis-platinum (cis-Pt)-induced HIV expression. While salicylic acid treatment, indomethacin treatment, UV exposure, or cis-Pt treatment alone decreased viability by up to 50%, equal numbers of viable cells were used for the CAT assays. Repression was evident if salicylic acid was administered 2 h before, at the same time as, or up to 6 h after exposure to the DNA-damaging agent. The kinetics were similar for UV- and for cis-Pt-induced HIV expression, and induction was dependent on the UV dose or cis-Pt concentration added to the culture. pH changes of the media alone in the absence of salicylic acid did not affect HIV expression. Indomethacin (100 microM) did not affect UV- or cis-Pt-induced HIV expression. These results suggest a role for the prostaglandins or the cyclo-oxygenase pathway or both in HIV induction mediated by DNA-damaging agents.

Identification of the dominant photochemical pathways and mechanistic insights to the ultrafast ligand exchange of Fe(CO)5 to Fe(CO)4EtOH.

We utilized femtosecond time-resolved resonant inelastic X-ray scattering and ab initio theory to study the transient electronic structure and the photoinduced molecular dynamics of a model metal carbonyl photocatalyst Fe(CO)5 in ethanol solution. We propose mechanistic explanation for the parallel ultrafast intra-molecular spin crossover and ligation of the Fe(CO)4 which are observed following a charge transfer photoexcitation of Fe(CO)5 as reported in our previous study [Wernet et al., Nature 520, 78 (2015)]. We find that branching of the reaction pathway likely happens in the (1)A1 state of Fe(CO)4. A sub-picosecond time constant of the spin crossover from (1)B2 to (3)B2 is rationalized by the proposed (1)B2 → (1)A1 → (3)B2 mechanism. Ultrafast ligation of the (1)B2 Fe(CO)4 state is significantly faster than the spin-forbidden and diffusion limited ligation process occurring from the (3)B2 Fe(CO)4 ground state that has been observed in the previous studies. We propose that the ultrafast ligation occurs via (1)B2 → (1)A1 → (1)A' Fe(CO)4EtOH pathway and the time scale of the (1)A1 Fe(CO)4 state ligation is governed by the solute-solvent collision frequency. Our study emphasizes the importance of understanding the interaction of molecular excited states with the surrounding environment to explain the relaxation pathways of photoexcited metal carbonyls in solution.

Human complement protein C8 gamma.

Human C8 gamma is a 22 kDa subunit of complement component C8, which is one of five components (C5b, C6, C7, C8, C9) that interact to form the cytolytic membrane attack complex (MAC) of complement. C8 contains three nonidentical subunits (alpha, beta, gamma) that are products of different genes. These subunits are arranged asymmetrically to form a disulfide-linked C8 alpha-gamma dimer that is noncovalently associated with C8 beta. C8 alpha and C8 beta are homologous to C6, C7 and C9 and together these proteins comprise what is referred to as the 'MAC protein family'. By comparison, C8 gamma is distinct in that it belongs to the lipocalin family of small, secreted proteins which have the common ability to bind small hydrophobic ligands. While specific roles have been identified for C8 alpha and C8 beta in the formation and function of the MAC, a function for C8 gamma and the identity of its ligand are unknown. This review summarizes the current status of C8 gamma structure and function and the progress made from efforts to determine its role in the complement system.

HIV expression is induced in dying cells.

Using HeLa cells stably transfected with an HIV-LTR-CAT construct, we demonstrated a peak in CAT induction that occurs in viable (but not necessarily cell-division-competent) cells 24 h following exposure to some cell-killing agents. gamma rays were the only cell-killing agent which did not induce HIV transcription; this can be attributed to the fact that gamma-ray-induced apoptotic death requires functional p53, which is not present in HeLa cells. For all other agents, HIV-LTR induction was dose-dependent and correlated with the amount of cell killing that occurred in the culture. Doses which caused over 99% cell killing induced HIV-LTR transcription maximally, demonstrating that cells that will go on to die by 14 days are the cells expressing HIV-LTR-CAT.

Effects of gamma rays, ultraviolet radiation, sunlight, microwaves and electromagnetic fields on gene expression mediated by human immunodeficiency virus promoter.

Previous work by our group and others has shown the modulation of human immunodeficiency virus (HIV) promoter or long terminal repeat (LTR) after exposure to neutrons and ultraviolet radiations. Using HeLa cells stably transfected with a construct containing the chloramphenicol acetyl transferase (CAT) gene, the transcription of which is mediated by the HIV-LTR, we designed experiments to examine the effects of exposure to different types of radiation (such as gamma rays, ultraviolet and sunlight irradiations, electromagnetic fields and microwaves) on HIV-LTR-driven expression of CAT. These results demonstrated ultraviolet-light-induced transcription from the HIV promoter, as has been shown by others. Exposure to other DNA-damaging agents such as gamma rays and sunlight (with limited exposures) had no significant effect on transcription mediated by HIV-LTR, suggesting that induction of HIV is not mediated by just any type of DNA damage but rather may require specific types of DNA damage. Microwaves did not cause cell killing when cells in culture were exposed in high volumes of medium, and the same cells showed no changes in expression. When microwave exposure was carried out in low volumes of medium (so that excessive heat was generated) induction of HIV-LTR transcription (as assayed by CAT activity) was evident. Electromagnetic field exposures had no effect on expression of HIV-LTR. These results demonstrate that not all types of radiation and not all DNA-damaging agents are capable of inducing HIV. We hypothesize that induction of HIV transcription may be mediated by several different signals after exposure to radiation.

Correlations between flow resistance and geometry in a model of the human nose.

The relationship between the pressure losses within the nasal airways and nasal geometry were studied in a 3:1 scale model. The geometry of the model was based on magnetic resonance images of the skull of a healthy male subject. Pressure measurements, flow visualization, and hot-wire anemometry studies were performed at flow rates that, in vivo, corresponded to flows of between 0.05 and 1.50 l/s. The influence of nasal congestion and the collapse of the external nares were examined by using modeling clay to simulate local constrictions in the cross section. A dimensionless analysis of the pressure losses within three sections of the airway revealed the influence of various anatomic dimensions on nasal resistance. The region of the exterior nose behaves as a contraction-expansion nozzle in which the pressure losses are a function of the smallest cross-sectional area. Losses in the interior nose resemble those associated with channel flow. The nasopharynx is modeled as a sharp bend in a circular duct. Good correspondence was found between the predicted and actual pressure losses in the model under conditions that stimulated local obstructions and congestion.

The effects of multiple UV exposures on HIV-LTR expression.

Previous studies have shown that cellular stress agents such as UV radiation induce transcription from the long terminal repeat (LTR) of the human immunodeficiency virus (HIV). Using HeLa cells stably transfected with the HIV-LTR sequence, which transcriptionally drives the chloramphenicol acetyl transferase (CAT) reporter gene, we examined the effects of multiple exposures to UVC (254 nm) on HIV-LTR-CAT expression. Low doses (< or = 5 J m-2) had no effect on CAT expression, but up to 29-fold induction was observed with 10 J m-2 when cells were harvested 48 h after completion of the exposure. Little difference was noted in induction levels when cells were exposed to one 25 J m-2 dose, viable cells were harvested at 24 h, 48 h or 72 h, and cell lysates were assayed for CAT expression. Two sequential 12.5 J m-2 exposures, given 24 h apart, resulted in an additive effect on CAT expression; these two exposures produced CAT activity equivalent to that induced following a single 25 J m-2 dose. This additive effect was not evident at the lower doses (< or = 5 J m-2) or at the higher doses. Maximal induction was observed using doses from 25 to 37.5 J m-2. Multiple exposures with either the low (< or = 5 J m-2) or high doses (> 25 J m-2) did not result in an additive effect.(ABSTRACT TRUNCATED AT 250 WORDS)

In vitro calcification of pericardial bioprostheses.

BACKGROUND AND AIM OF THE STUDY: The lifetime of bioprosthetic heart valves is limited by calcification. To investigate the calcification behavior of bioprostheses and gain insight into the etiology of valve calcification, a test protocol for accelerated valve calcification was developed. This protocol includes a pulsatile valve tester, a synthetic calcification fluid, and non-destructive radiographic assessment of calcification sites. About 40 porcine bioprostheses from different manufacturers have been investigated previously using this test protocol and showed that valves exhibited different calcification patterns and even different degrees of calcification within their leaflets. A positive correlation of calcification versus tissue anomalies/stress concentrations (r = 0.72; n = 29 valves) and lipid deposits (r = 0.81) was found. In the present study, bovine pericardial valves were investigated in comparison with porcine valves. METHODS: Four bovine pericardial and two porcine mitral valves (Baxter) with a tissue annulus diameter (TAD) of 29 mm (one 27 mm) were investigated in parallel under identical test conditions. The valves were cyclically loaded at 300 per min with a delta p of 110 mmHg at 37 degrees C for up to 19 x 10(6) cycles. The synthetic calcification fluid was changed weekly. Sites of calcification were assessed by microradiography. Radiographs were analyzed by PC images processing with respect to the degree of calcification, defined as calcified surface area in relation to total leaflet surface area. RESULTS: This analysis showed that, for bovine pericardial valves, the mean degree of calcification increased by 14% and 20% after 12 and 19 x 10(6) cycles, respectively. Under identical conditions, the mean degree of calcification of porcine valves increased by 28% and 37%. CONCLUSIONS: Pericardial valves appear less prone to calcification than porcine valves. Further studies must be performed in order to prove this finding since, as recognized previously in porcine valves, other factors such as tissue or manufacturing anomalies may be as important as the tissue source itself.

The effect of left ventricular dP/dt on the in vitro dynamics of the Björk-Shiley Convexo-Concave mitral valve.

Left ventricular (LV) dP/dt is considered an important hemodynamic factor influencing the dynamics of mechanical heart valve prostheses. LV dP/dt is dependent on patient factors including age, cardiac activity, health, and medication. The objective of this study was to determine the effect of LV dP/dt on the closing dynamics of mechanical heart valve prostheses in the mitral position. Eight instrumented 29 mm Björk-Shiley Convexo-Concave (BSCC) heart valves were tested in the pulse duplicator of the Helmholtz Institute. The valves had miniature strain gages mounted at the base of the outlet strut to measure impact loads at closure. Closing velocities were measured with a "light gate" device which was triggered by the closing leaflet. Physiologic pressure and flow waveforms were generated by a computer-controlled hydraulic drive unit. LV dP/dt was varied from 500 to 4000 mmHg/s simulating a wide range of physiologic conditions. It was found that the closing velocity was almost linearly related to LV dP/dt. At 4000 mmHg/s, closing velocities ranged from 1.5 to 2.0 m/s. Impact loads increased monotonically with LV dP/dt and closing velocity. In some valves, impact loads reached 2800 g at LV dP/dt of 4000 mmHg/s, and closing velocities of 2.0 m/s.

Pressure distribution near the occluders and impact forces on the outlet struts of Björk-Shiley convexo-concave valves during closing.

BACKGROUND AND AIMS OF THE STUDY: An in vitro study of the mechanics of closure of Björk-Shiley convexo-concave (BSCC) valves is presented in order to investigate the mechanics of outlet strut fracture reported in a small fraction of the implanted valves. MATERIALS AND METHODS: Four BSCC 29 mm valves instrumented with strain gages on the outlet strut legs were mounted in the mitral position of an axisymmetric flow chamber of a mock pulsatile flow loop. Measurements of the pressure field in the vicinity of the occluder, closing velocity of the occluder tip in the major orifice, and the impact force between the occluder and outlet strut at the instant of valve closure were obtained at a range of physiologic flow rates. RESULTS: The results indicated an uneven pressure distribution on the occluder associated with a tendency for the occluder to over-rotate and induce loads on the outlet struts. The impact loads on the outlet struts were asymmetric with load on one leg being larger than the other by up to 25%. These results are consistent with single leg separation preceding outlet strut fracture in most of the valve failures reported. Orientation of the valve with respect to the mitral orifice (major orifice towards the top or bottom) did not significantly affect the loads on the outlet strut. A significant variation in the impact loads of the four valves was measured for identical experimental conditions suggesting that valve specific factors influence outlet strut loads. CONCLUSIONS: This study provided an understanding of the cause-effect relationship between valve dynamics and outlet strut fracture.

Dynamics of Björk-Shiley Convexo-Concave mitral valves in sheep.

The outlet strut of Björk-Shiley (BSCC) Convexo-Concave heart valves has fractured in some implanted valves resulting in disc escape and emergency reoperation or death. The closing dynamics of BSCC heart valves was studied in situ to determine the forces acting on the outlet strut during valve closure. BSCC valves with strain gages attached to the outlet strut were implanted in the mitral position in sheep. Impact forces were measured under various circumstances including anesthetized, sedentary, and exercise conditions. Left ventricular (LV) pressures were recorded simultaneously via catheter-tip pressure transducers. Impact forces increased with the 1/3 power of left ventricular pressure rise and the square root of LV pressure at valve closure. Good agreement between the in vivo measurements and theoretical models was observed. Based on this study, it is concluded that sheep provides a good in situ model for the testing of mechanical mitral valves.

Velocity of closure of Björk-Shiley Convexo-Concave mitral valves: effect of mitral annulus orientation and rate of left ventricular pressure rise.

The purpose of this study was to determine analytically the hemodynamic factors that affect the closing velocity of the disc of Björk-Shiley convexo-concave (BSCC) prosthetic mitral valves. The motion of the BSCC disk was modelled by Newton's second law written in the form of a second order differential equation which expressed the instantaneous angle of the disc with respect to the valve ring as a function of the instantaneous pressure drop across the mitral valve, delta P(t), and the angle of the pressure gradient vector acting upon the disc during closure. The disc closes in response to the negative pressure drop created by the crossover of left atrial and left ventricular (LV) pressures. The rate of closure depends on the rate of development of the pressure drop across the valve, d delta P/dt, which is largely dependent upon the rate of change of left ventricular pressure during isovolumic contraction, LV dP/dt. The closure rate is also strongly dependent on the initial angle of the pressure drop vector with respect to the disc. The disc was predicted to reach its highest velocity at the moment of impact, based on the Runge-Kutta solution. Modelling suggests that a high LV dP/dt during valve closure or distorted LV geometry, causing the angle between the fully open disc and the pressure drop vector to shift, will cause the valve to have a high velocity at the moment of impact and may produce high impact loads.

Strut fracture mechanisms of the Björk-Shiley convexo-concave heart valve.

Investigations of convexo-concave (C/C) valve outlet strut fractures (OSFs) were initially confounded by knowledge that the strut was subject to bending forces in arresting the opening disc. Pulse duplicator studies subsequently showed that closing loads were all born by the inlet strut, along with an understandable focus on the nature of the welds, where most fractures occurred. As observations of explanted valves accumulated, certain features pointed to unusual closing loads that might be contributory factors, but these hypothetical forces could not be verified. Epidemiological extrapolations and case-matched control studies have shown that certain valve and patient characteristics were each associated independently with increased OSF risk, leading to clinically valuable risk stratification, but little additional understanding of why OSFs continued to occur. Detection of the causative, highly transient (< 0.5 ms), outlet-strut-tip impacts due to closing disc over-rotation that have almost ten times the force of disc opening, and the capability of inducing leg-base bending stresses beyond the strut wire's fatigue endurance limit had to await the development of computer-controlled pulse duplicators and strut-leg strain gaging. Exercised young animals easily achieved such strut loading, but most human patients would probably have more difficulty. The actual OSF mechanism is a long-term, valve-patient interaction that requires the concurrence of susceptible valve geometry and sufficient ventricular contractility potential to develop the isovolumic, high dP/dt needed for forceful disc over-rotation. Critical strut tip loading must then occur often enough to fatigue fracture both strut legs within the patient's lifetime with the valve.