Soft X-ray microscopy for probing of topical tacrolimus delivery via micelles.

The penetration of topically applied tacrolimus formulated in micelles into murine skin is reported, measured by X-ray microscopy. Tacrolimus and micelles are probed for the first time by this high spatial resolution technique by element-selective excitation in the C 1s- and O 1s-regimes. This method allows selective detection of the distribution and penetration depth of drugs and carrier molecules into biologic tissues. It is observed that small, but distinct quantities of the drug and micelles, acting as a drug carrier, penetrate the stratum corneum. A comparison is made with the paraffin-based commercial tacrolimus ointment Protopic®, where local drug concentrations show to be low. A slight increase in local drug concentration in the stratum corneum is observed, if tacrolimus is formulated in micelles, as compared to Protopic®. This underscores the importance of the drug formulations for effective drug delivery. Time-resolved penetration shows presence of drug in the stratum corneum 100 min after formulation application, with penetration to deeper skin layers at 1000 min. High resolution micrographs give indications for a penetration pathway along the lipid membranes between corneocytes, but also suggest that the compound may penetrate corneocytes.

Influence of the skin barrier on the penetration of topically-applied dexamethasone probed by soft X-ray spectromicroscopy.

The penetration of dexamethasone into human skin ex vivo is reported. X-ray microscopy is used for label-free probing of the drug and quantification of the local drug concentration with a spatial resolution reaching 70±5nm. This is accomplished by selective probing the dexamethasone by X-ray absorption. Varying the penetration time between 10min and 1000min provides detailed information on the penetration process. In addition, the stratum corneum has been damaged by tape-stripping in order to determine the importance of this barrier regarding temporally resolved drug penetration profiles. Dexamethasone concentrations distinctly vary, especially close to the border of the stratum corneum and the viable epidermis, where a local minimum in drug concentration is observed. Furthermore, near the basal membrane the drug concentration strongly drops. High spatial resolution studies along with a de-convolution procedure reveal the spatial distribution of dexamethasone in the interspaces between the corneocytes consisting of stratum corneum lipids. These results on local drug concentrations are interpreted in terms of barriers affecting the drug penetration in human skin.

Core-multishell nanocarriers: Transport and release of dexamethasone probed by soft X-ray spectromicroscopy.

Label-free detection of core-multishell (CMS) nanocarriers and the anti-inflammatory drug dexamethasone is reported. Selective excitation by tunable soft X-rays in the O 1s-regime is used for probing either the CMS nanocarrier or the drug. Furthermore, the drug loading efficiency into CMS nanocarriers is determined by X-ray spectroscopy. The drug-loaded nanocarriers were topically applied to human skin explants providing insights into the penetration and drug release processes. It is shown that the core-multishell nanocarriers remain in the stratum corneum when applied for 100min to 1000min. Dexamethasone, if applied topically to human ex vivo skin explants using different formulations, shows a vehicle-dependent penetration behavior. Highest local drug concentrations are found in the stratum corneum as well as in the viable epidermis. If the drug is loaded to core-multishell nanocarriers, the concentration of the free drug is low in the stratum corneum and is enhanced in the viable epidermis as compared to other drug formulations. The present results provide insights into the penetration of drug nanocarriers as well as the mechanisms of controlled drug release from CMS nanocarriers in human skin. They are also compared to related work using dye-labeled nanocarriers and dyes that were used as model drugs.

Selective Probing of the Penetration of Dexamethasone into Human Skin by Soft X-ray Spectromicroscopy.

Selective probing of dexamethasone in excised human skin using soft X-ray spectromicroscopy provides quantitative concentration profiles as well as two-dimensional drug distribution maps. Element- and site-selective excitation of dexamethasone at the oxygen K-edge with the lateral step width adjusted to 1 µm provides detailed information on the location of the drug in the different skin layers. The key of this work is to probe dexamethasone selectively at the carbonyl site (C3) by the O 1s → π* transition, providing also a most efficient way to quantify the drug concentration as a function of penetration depth in correlation with structural properties of the skin containing carboxyl and amide oxygen sites occurring at higher transition energy than dexamethasone. Following drug exposure for 4 h, the glucocorticoide is located in about equal amounts in the stratum corneum, the outermost horny layer of skin, and in the viable epidermis, whereas in the dermis no dexamethasone is detected. In the stratum corneum, most of the lipophilic drug is found in regions between corneocytes, where epidermal lipids are dominating.

Gas-to-solid shift of C 1s-excited benzene.

The gas-to-solid shift of benzene is reported in the C 1s-core level regime, where the C 1s → π*-transition is investigated between 284.0 eV and 286.5 eV. Simultaneous experiments on the gas phase and condensed species are used to determine the gas-to-solid shift within an accuracy of ±5 meV. Specifically, it is observed that the vibrationally resolved C 1s → π*-transition in solid benzene is red-shifted by 55 ± 5 meV relative to the transition of the isolated molecule. Contrary to previously reported experimental data and estimates this gas-to-solid shift is somewhat smaller than the gas-to-cluster shift. It is significantly smaller than that determined in previous work on gaseous and condensed benzene. These results are discussed in terms of structural properties of molecular clusters and solid benzene by involving ab initio calculations as well as processes leading to spectral shifts of core-excited variable size matter. Finally, changes in the shape of the C 1s → π*-band upon the formation of solid benzene and benzene clusters are discussed.

Site-dependent spectral shifts in core-to-pi* excitations of pyridine clusters.

Site- and element-selective core-to-pi* excitation in free pyridine clusters is investigated. The experimental results indicate the occurrence of site- and size-dependent spectral shifts in the C 1s and N 1s --> pi* excitation regime. Specifically, we observe in the C 1s regime a substantial and site-dependent redshift of the low energy slopes of the C 1s --> pi* band by 90 meV in clusters relative to the bare molecule, whereas the high energy slopes of this band remain almost unchanged. In contrast, a size-dependent blueshift of the same order of magnitude is found for the entire N 1s --> pi* band. This is distinctly different from previous results on van der Waals clusters, where exclusively redshifts in 1s --> pi* transitions are observed. The experimental results are compared to ab initio calculations, which serve to simulate the 1s --> pi*( v = 0) transitions. These results clearly indicate that the spectral shifts are primarily a result of electrostatic interactions between the molecular moieties and that an antiparallel orientation of molecular units preferably dominates in variable-size pyridine clusters.

Inner-valence photoionization of O((1)D): experimental evidence for the 2s(2)2p(4)((1)D)-->2s(1)2p(5)((1)P) transition.

Photoionization and autoionization of electronically excited atomic oxygen O((1)D) are investigated in the energy range between 12 and 26 eV using tunable laser-produced plasma radiation in combination with time-of-flight mass spectrometry. A broad, asymmetric, and intense feature is observed that is peaking at 20.53+/-0.05 eV. It is assigned to the 2s(2)2p(4)((1)D)-->2s(1)2p(5)((1)P) transition, which subsequently autoionizes by a Coster-Kronig transition, as predicted by the previous theoretical work [K. L. Bell et al., J. Phys. B 22, 3197 (1989)]. Specifically, the energy of the unperturbed transition occurs at 20.35+/-0.07 eV. Its shape is described by a Fano profile revealing a q parameter of 4.25+/-0.8 and a width of gamma=2.2+/-0.15 eV. Absolute photoionization cross section sigma is derived, yielding sigma=22.5+/-2.3 Mb at the maximum of the resonance. In addition, weak contributions to the O((1)D) yield from dissociative ionization originating from molecular singlet oxygen [O(2)((1)Delta(g))] are identified as well. Possible applications of the 2s(2)2p(4)((1)D)-->2s(1)2p(5)((1)P) transition as a state-selective and sensitive probe of excited oxygen in combination with photoionization mass spectrometry are briefly discussed.

Elastic light scattering from free sub-micron particles in the soft X-ray regime.

We report the first experimental results on angle-resolved elastic light scattering in the soft X-ray regime, where free sub-micron particles in the size regime between 150 and 250 nm are studied in the gas phase by using a continuous particle beam. Two different types of studies are reported: (i) Angle-resolved elastic light scattering experiments provide specific information on the scattering patterns in the regime of element-selective inner-shell excitation near the Si 2p-edge (80-150 eV). In addition to intense forward scattering, we observe distinct features in the angle-resolved scattering patterns. These are modelled by using Mie theory as well as a model that includes contributions from diffuse and specular reflection. The results are primarily attributed to scattering from soft X-rays in the surface layer. (ii) Spectroscopic experiments are reported, where the photon detector is placed at a given scattering angle while scanning the photon energy near the Si 2p-absorption edge. These results are also analyzed by a Mie model, yielding accurate information of the size distribution.

Radiative relaxation and fragmentation dynamics of S 2p-excited hydrogen sulfide.

Radiative relaxation of S 2p-excited hydrogen sulfide (H(2)S) is investigated by dispersed ultraviolet and visible fluorescence spectroscopies. We observe distinct changes in the fluorescence spectra as a function of excitation energy. Excitation to Rydberg states below the S 2p ionization threshold yields intense fluorescence from neutral and ionic atomic fragments (H, S(+), and S(2+)). In addition to the atomic emission, fluorescence of the molecular fragment ion HS(+) is preferably found after excitation of the S 2p electron into the unoccupied 6a(1) and 3b(2) orbitals with sigma(*) character. This is interpreted as evidence for ultrafast dissociation of the core-excited molecule prior to electronic relaxation. The rotationally resolved fluorescence spectra of the A (3)Pi-->X (3)Sigma(-) transition are analyzed in terms of the fragmentation dynamics leading to the formation of the excited molecular fragment ion, where changes in bond angle are discussed in terms of the rotational population.

Charging mechanisms of trapped element-selectively excited nanoparticles exposed to soft x rays.

Charging mechanisms of trapped, element-selectively excited free SiO2 nanoparticles by soft x rays are reported. The absolute charge state of the particles is measured and the electron emission probability is derived. Changes in electron emission processes as a function of photon energy and particle charge are obtained from the charging current. This allows us to distinguish contributions from primary photoelectrons, Auger electrons, and secondary electrons. Processes leading to no change in charge state after absorption of x-ray photons are identified. O 1s-excited SiO2 particles of low charge state indicate that the charging current follows the inner-shell absorption. In contrast, highly charged SiO2 nanoparticles are efficiently charged by resonant Auger processes, whereas direct photoemission and normal Auger processes do not contribute to changes in particle charge. These results are discussed in terms of an electrostatic model.

C 1s -->pi* excitation in variable size benzene clusters.

The C 1s -->pi* transition in molecular benzene and benzene clusters is investigated by photoion yields at high energy resolution. The vibrationally resolved band shows the same shape in clusters as in the bare molecule, but it is redshifted by 50 meV in small clusters, i.e. near the threshold of cluster formation. This redshift increases to 70 meV with increasing cluster size. The results are assigned in comparison with ab initio calculations on model structures of dimers, trimers, and tetramers. These indicate that different carbon sites in the molecular moieties give rise to distinct spectral shifts, where carbon sites that are pointing to the pi-system of another molecule show a larger redshift than the other ones. Such structural properties are found in solid benzene, so that the gas-to-solid shift of C 1s -->pi* excited benzene is derived to be a redshift which is of the order of 100-180 meV.

Shape resonances in molecular clusters: the 2t(2g) shape resonances in S 2p-excited sulfur hexafluoride clusters.

Sulfur hexafluoride clusters are investigated in the S 2p excitation regime. For the first time special emphasis is put on high-energy resolution spectroscopy of shape resonances in free clusters. We have investigated the 2t(2g)-shape resonance occurring above the S 2p threshold as one typical example to study size effects that are related to shape resonances. A small redshift of this resonance of 30 +/- 5 meV occurs in clusters relative to the free molecule and changes in line shape are observed. A double-barrier optical potential model is applied for the analysis of intra- and intermolecular effects occurring in SF6 clusters, which is suitable for rationalizing the experimentally observed spectral changes. The experimental and theoretical results are briefly discussed in comparison to previous work on core-excited van der Waals clusters containing diatomic molecules.

Bond strength of chlorine peroxide.

The bond strength of chlorine peroxide (ClOOCl) is studied by photoionization mass spectrometry. The experimental results are obtained from the fragmentation threshold yielding ClO+, which is observed at 11.52 +/- 0.025 eV. The O-O bond strength D(o) is derived from this value in comparison to the first ionization energy of ClO, yielding D(o)298 = 72.39 +/- 2.8 kJ mol(-1). The present work provides a new and independent method to examine the equilibrium constant K(eq) for chlorine peroxide formation via dimerization of ClO in the stratosphere. This yields an approximation for the equilibrium constant in the stratospheric temperature regime between 190 and 230 K of the form K(eq) = 1.92 x 10(-27) cm3 molecules(-1) x exp(8430 K/T). This value of K(eq) is lower than current reference data and agrees well with high altitude aircraft measurements within their scattering range. Considering the error limits of the present experimental results and the resulting equilibrium constant, there is agreement with previous works, but the upper limit of current reference values appears to be too high. This result is discussed along with possible atmospheric implications.

Cluster size effects in core excitons of 1s-excited nitrogen.

Cluster size effects in core excitons below the N 1s ionization energy of nitrogen clusters are reported in the energy regime 405-410 eV. These results are compared to the molecular Rydberg states as well as the corresponding bulk excitons of condensed nitrogen. The experimental results are assigned using ab initio calculations. It is found that the lowest excitons (N 1s-->3ssigma and N 1s-->3ppi) are blueshifted relative to the molecular Rydberg transitions, whereas others (N 1s-->3dpi and N 1s-->4ppi) show a redshift. Results from ab initio calculations on (N(2))(13) clearly indicate that the molecular orientation within a cluster is critical to the spectral shift, where bulk sites as well as inner- and outer-surface sites are characterized by different inner-shell absorption energies. These results are compared to the experimental spectra as well as previous work on site-selectively excited atomic van der Waals clusters, providing an improved spectral assignment of core exciton states in weakly bound molecular clusters and the corresponding condensed phase.

Mechanism of anion formation in C 1s-->pi*-excited carbon dioxide.

The mechanism of anion formation from core-excited carbon dioxide is investigated in the C 1s-excitation regime (280-340 eV), where negative-ion-positive-ion coincidences and negative-ion-positive-ion-positive-ion coincidences experiments are performed. O(-) formation occurs efficiently upon C 1s-->pi(*)-excitation (290.7 eV) from the singly charged cation CO(2) (+). This anion is measured in coincidence with the singly charged atoms C(+) and O(+). The formation of atomic cations is accompanied by a substantial kinetic energy release. This suggests that an intermediate CO(++) is formed together with O(-), where the dication decays via fission. The results are discussed in terms of an anion formation mechanism from core-excited molecules considering previous experimental results.

Charge localization in collision-induced multiple ionization of van der Waals clusters with highly charged ions.

Charge localization in multiple ionization and fragmentation of small argon clusters is reported. The processes are initiated by interaction of the neutral cluster with highly charged Xe(q+) (5< or =q< or =25). Products are detected by means of multicoincidence time-of-flight methods. A strong dependence of the fragmentation pattern on the Xe charge state q is observed. In particular, we find evidence for formation of multiply charged atomic Ar(r+) fragment ions up to r = 7. Such high charge states have neither been observed in fission of multiply charged van der Waals clusters nor in ion-induced fragmentation of fullerenes or metal clusters. This hints at fundamentally different excitation and fragmentation dynamics.

Progress on inner-shell excitations of molecular van der Waals clusters.

Recent progress on core-level excitation of molecular van der Waals clusters is reported. Resonant excitation near element K edges of isolated and clustered molecules gives rise to small spectral shifts that can only be detected with high spectral resolution. This requires the use of state-of-the-art storage-ring facilities along with insertion devices and high-resolution soft X-ray monochromators. Selected experimental results on carbon monoxide clusters are reported. For the vibrationally resolved C 1s --> pi* (v = 0) band of clustered CO, these indicate characteristic line broadening as well as a small red shift of 2 +/- 1 meV compared with the isolated molecule. The results are discussed within the framework of the quasi-atomic approach with respect to intermolecular interactions, freezing of molecular rotations in clusters, and dynamic localization of resonant core-to-valence excitations.

Dynamic stabilization in 1sigma(u)-->1pi(g) excited nitrogen clusters.

High-resolution 1s near-edge spectra of molecular nitrogen and variable size nitrogen clusters obtained using monochromatic synchrotron radiation from the high brilliance BESSY-II storage ring facility are reported. The vibrationally resolved 1sigma(u)-->1pi(g) core-to-valence excitation band of clusters shows a distinct redshift of 6+/-1 meV relative to the isolated molecule, but the vibrational structure and linewidths are essentially unchanged. This shift is assigned to dynamic stabilization of 1sigma(u)-->1pi(g) excited molecules in clusters, arising from the dynamic dipole moment generated by core-hole localization in the low-symmetry cluster field. This leads to changes in intermolecular interactions compared to the ground-state cluster. Such spectral shifts are expected to occur generally in molecular clusters and in the corresponding condensed phase.

Acute bleeding caused by rupture of the thyroid gland following blunt neck trauma: case report.

We describe a case of blunt neck trauma that resulted in extensive rupture of a normal thyroid gland. Our experience shows that lesions of the thyroid gland must be taken into diagnostic consideration after blunt neck trauma, even in the absence of obvious signs of injury.