Biomedical applications of the ESRF synchrotron-based microspectroscopy platform.

Very little is known about the sub-cellular distribution of metal ions in cells. Some metals such as zinc, copper and iron are essential and play an important role in the cell metabolism. Dysfunctions in this delicate housekeeping may be at the origin of major diseases. There is also a prevalent use of metals in a wide range of diagnostic agents and drugs for the diagnosis or treatment of a variety of disorders. This is becoming more and more of a concern in the field of nanomedicine with the increasing development and use of nanoparticles, which are suspected of causing adverse effects on cells and organ tissues. Synchrotron-based X-ray and Fourier-transformed infrared microspectroscopies are developing into well-suited sub-micrometer analytical tools for addressing new problems when studying the role of metals in biology. As a complementary tool to optical and electron microscopes, developments and studies have demonstrated the unique capabilities of multi-keV microscopy: namely, an ultra-low detection limit, large penetration depth, chemical sensitivity and three-dimensional imaging capabilities. More recently, the capabilities have been extended towards sub-100nm lateral resolutions, thus enabling sub-cellular chemical imaging. Possibilities offered by these techniques in the biomedical field are described through examples of applications performed at the ESRF synchrotron-based microspectroscopy platform (ID21 and ID22 beamlines).

Photon-based techniques for nondestructive subsurface analysis of painted cultural heritage artifacts.

Often, just micrometers below a painting's surface lies a wealth of information, both with Old Masters such as Peter Paul Rubens and Rembrandt van Rijn and with more recent artists of great renown such as Vincent Van Gogh and James Ensor. Subsurface layers may include underdrawing, underpainting, and alterations, and in a growing number of cases conservators have discovered abandoned compositions on paintings, illustrating artists' practice of reusing a canvas or panel. The standard methods for studying the inner structure of cultural heritage (CH) artifacts are infrared reflectography and X-ray radiography, techniques that are optionally complemented with the microscopic analysis of cross-sectioned samples. These methods have limitations, but recently, a number of fundamentally new approaches for fully imaging the buildup of hidden paint layers and other complex three-dimensional (3D) substructures have been put into practice. In this Account, we discuss these developments and their recent practical application with CH artifacts. We begin with a tabular summary of 14 IR- and X-ray-based imaging methods and then continue with a discussion of each technique, illustrating CH applications with specific case studies. X-ray-based tomographic and laminographic techniques can be used to generate 3D renditions of artifacts of varying dimensions. These methods are proving invaluable for exploring inner structures, identifying the conservation state, and postulating the original manufacturing technology of metallic and other sculptures. In the analysis of paint layers, terahertz time-domain spectroscopy (THz-TDS) can highlight interfaces between layers in a stratigraphic buildup, whereas macrosopic scanning X-ray fluorescence (MA-XRF) has been employed to measure the distribution of pigments within these layers. This combination of innovative methods provides topographic and color information about the micrometer depth scale, allowing us to look "into" paintings in an entirely new manner. Over the past five years, several new variants of traditional IR- and X-ray-based imaging methods have been implemented by conservators and museums, and the first reports have begun to emerge in the primary research literature. Applying these state-of-the-art techniques in a complementary fashion affords a more comprehensive view of paintings and other artworks.

Wavelength-dispersive spectrometer for X-ray microfluorescence analysis at the X-ray Microscopy beamline ID21 (ESRF).

The development of a wavelength-dispersive spectrometer for microfluorescence analysis at the X-ray Microscopy ID21 beamline of the European Synchrotron Radiation Facility (ESRF) is reported. The spectrometer is based on a polycapillary optic for X-ray fluorescence collection and is operated in a flat-crystal geometry. The design considerations as well as operation characteristics of the spectrometer are presented. The achieved performances, in particular the energy resolution, are compared with the results of Monte Carlo simulations. Further improvement in the energy resolution, down to approximately eV range, by employing a double-crystal geometry is examined. Finally, examples of applications requiring both spatial and spectral resolutions are presented.

The interpretation of sulfur K-edge XANES spectra: a case study on thiophenic and aliphatic sulfur compounds.

Sulfur K-edge XANES has been measured for three sulfur model compounds, dibenzothiophene, dibenzothiophene sulfone, and aliphatic sulfur (DL-methionine). The spectra have been simulated with Density Functional Theory (DFT) by using a number of methods, including the half-core-hole approximation. Dipole transition elements were calculated and the transitions were convoluted with linearly increasing Gaussian functions in the first 20 eV of the near-edge region. In the case of dibenzothiophene, relaxation of the first excited states in the presence of the core-hole gave a further improvement. The theoretical results reproduce well the features of the spectra and give insight in the relation between geometric structure and molecular orbitals. Though DL-methionine and dibenzothiophene show a similar sharp rise of the white line, their molecular levels are quite different, pointing out the difficulties in finding useful "fingerprints" in the spectra for specific compounds.

X-ray absorption near-edge structure of GaN with high Mn concentration grown on SiC.

By means of x-ray absorption near-edge structure (XANES) several Ga(1-x)Mn(x)N (0.03

New cryogenic environment for beamline ID22 at the European Synchrotron Radiation Facility.

A compact minicryostat has been well adapted on the hard x-ray microprobe ID22 of the European Synchrotron Radiation Facility. For variable low-temperature investigations, its special technical design provides precise scanning microscopy and allows easy access for multiple detection modes. Based on x-ray excited optical luminescence technique on the micrometer scale, details of the equipment, its temperature calibration, and typical results are described. Data collections from InAs quantum heterostructures support the excellent thermal performance of the novel cryogenic device.

Micro-chemical imaging of cesium distribution in Arabidopsis thaliana plant and its interaction with potassium and essential trace elements.

Cesium as an alkali element exhibits a chemical reactivity similar to that of potassium, an essential element for plants. It has been suggested that Cs phytotoxicity might be due either to its competition with potassium to enter the plant, resulting in K starvation, or to its intracellular competition with K binding sites in cells. Such elemental interactions can be evidenced by chemical imaging, which determines the elemental distributions. In this study, the model plant Arabidopsis thaliana was exposed to 1 mM cesium in the presence (20 mM) or not of potassium. The quantitative imaging of Cs and endogenous elements (P, S, Cl, K, Ca, Mn, Fe, and Zn) was carried out using ion beam micro-chemical imaging with 5 microm spatial resolution. Chemical imaging was also evidenced by microfocused synchrotron-based X-ray fluorescence (microXRF) which presents a better lateral resolution (<1 microm) but is not quantitative. Cesium distribution was similar to potassium which suggests that Cs can compete with K binding sites in cells. Cesium and potassium were mainly concentrated in the vascular system of stems and leaves. Cs was also found in lower concentration in leaves mesophyll/epidermis. This late representing the larger proportion in mass, mesophyll/epidermis can be considered as the major storage site for cesium in A. thaliana. Trichomes were not found to accumulate cesium. Interestingly, increased Mn, Fe, and Zn concentrations were observed in leaves at high chlorosis. Mn and Fe increased more in the mesophyll than in veins, whereas zinc increased more in veins than in the mesophyll suggesting a tissue specific interaction of Cs with these trace elements homeostasis. This study illustrates the sensitivity of ion beam microprobe and microfocused synchrotron-based X-ray fluorescence to investigate concentrations and distributions of major and trace elements in plants. It also shows the suitability of these analytical imaging techniques to complement biochemical investigations of metallic stress in plants.

Evidence for various calcium sites in human hair shaft revealed by sub-micrometer X-ray fluorescence.

New information about calcium status in human scalp hair shaft, deduced from X-ray micro-fluorescence imaging, including its distribution over the hair section, the existence of one or several binding-types and its variation between people, is presented. The existence of two different calcium types is inferred. The first one corresponds to atoms (or ions) easily removable by hydrochloric acid, located in the cortex (granules), in the cuticle zone and also in the core of the medulla, which can reasonably be identified as calcium soaps. The second type consists of non-easily removable calcium atoms (or ions) that are located in the medulla wall, probably also in the cuticle, and rather uniformly in the cortex; these calcium atoms may be involved in Ca(2+)-binding proteins, and their concentration is fairly constant from one subject to another. In addition to its nonuniform distribution across the hair section, the second striking feature of the first type calcium content is its high variability from one subject to another, by up to a factor 10. We expect this information to be useful for analyzing in more detail the relationship between hair calcium and environmental and medical factors.

Deep brachial vein catheterization for total parenteral nutrition--an alternate approach: review of 154 cases.

The relative safety of silicon rubber catheters allows us to use deep brachial vein catheterization when other vascular accesses are hazardous or impossible. The vein is isolated from nerve and artery after a small incision is made across the inner edge of the brachial biceps. From January 1980 until April 1982, 154 catheters were inserted and remained in place from 14-167 days. In four instances the catheter had to be replaced because the tip was not in the superior vena cava. Twice bleeding had to be controlled with an occlusive bandage. We had no incidence of clinical thrombosis. The surgical access to the deep brachial vein provides a simple and safe method for difficult and even long-term vascular access.

[Antidepressive treatments. Predictive factors of tolerability]. / Traitements antidépresseurs. Facteurs prédictifs de la tolérance.

Conducting an antidepressant treatment supposes the analysis of various parameters concerning patients, depressive illness and psychotropic drugs. The first determinant of prescription is the expected antidepressant effect. But regarding the role of compliance in the efficacy of treatment, probable tolerability also plays a role. It remains difficult to predict tolerability. A questionnaire was designed to assess the relevance of various predictive factors taken into account in the prognosis of tolerability: global tolerability, patient's psychopathological structure, clinical aspects of depression and characteristics of the particular antidepressant being used.

[Cerebral x-ray computed tomography in anorexia nervosa. Study of 16 cases]. / Apport de la tomodensitométrie cérébrale dans l'anorexie mentale. Etude de 16 cas.

Cranial computerized tomography scan was used to study 16 cases of anorexia nervosa. The effect of treatment was explored by a second computerized tomography scan in 9 of the cases. Enlargement of cortical sulci and/or ventricular dilatation were demonstrated in 14 patients (87.5%). It was shown that the amelioration of body weight was related to the reversibility of computerized tomography scan abnormalities in 8 cases (89%). The authors discuss different aetiological hypotheses explaining these findings. They attempt to clear out the importance of the cranial computerized tomography in this disorder.

[Improvement of the postoperative period in surgery for tumors of the mouth floor by parenteral nutrition]. / Amélioration des suites opératoires des interventions pour tumeurs du plancher de la bouche grâce à la nutrition parentérale.

Since August 1977 total parenteral nutrition is part of post operative care for pull through operations for growth of floor of the mouth. The first 14 patients under total parenteral nutrition (from August 1977 until April 1980) are compared with the previous 14 patients. The length of the post operative period had slightly diminished but without statistical significance. On the other hand there is a significant difference for alimentary autonomy on discharge and number of complications (p less than 0.05). Our good handling of total parenteral nutrition has not replaced some complications by others.

Synchrotron-based X-ray spectromicroscopy used for the study of an atypical micrometric pigment in 16th century paintings.

Grünewald is a famous German painter of the 16th century, whose celebrity is associated with his unique skill in handling colors. This article presents the analysis of materials used to render a metallic aspect in the Isenhein Altarpiece and the Basel's Crucifixion. Such samples are challenging objects for microanalysis due to both chemical and physical complexity. Their study by synchrotron-based X-ray microscopy techniques was made possible thanks to recent developments carried out at the ID21 beam line (European Synchrotron Radiation Facility, ESRF). A submicron X-ray fluorescence probe revealed the main presence of lead, sulfur, antimony, and calcium. The fluorescence-line interferences (in particular K-lines of sulfur with M-lines of lead, and K-lines of calcium with L-lines of antimony) were resolved with the fitting program, PyMCA. 2D-mapping highlighted the presence of micrometer grains of sulfur and antimony into a lead matrix. XANES measurements were performed at both the sulfur K-edge and the antimony L-edge to refine information from an atomic to a molecular level. Beam stability was a key point in this study to selectively probe micrometer pigment grains, dispersed in the lead matrix. They confirm that the grains are made of stibnite (antimony sulfide), a very atypical pigment. Chemical mapping of sulfides is perfectly correlated with antimony mapping and provides a clear visualization of the stibnite pigments, in addition to their identification. Besides its artistic relevancy, this work aims at illustrating developments of synchrotron X-ray microprobe methods for the chemical characterization and observation of complex and micrometer-scale materials.

Energy-dispersive absorption spectroscopy for hard-X-ray micro-XAS applications.

Originally developed for time-resolved X-ray absorption spectroscopy (XAS), energy-dispersive absorption spectroscopy offers new opportunities for applications such as fluorescence detection and microbeams for scanning probe spectroscopy, thanks to recent developments in both instrumentation and optics. In this context, this paper presents a first example of chemical mapping recorded at ID24, the energy-dispersive XAS beamline at the ESRF. Attributes of this geometry for microanalysis are addressed. Finally, present and future plans are discussed and developed in the light of the evolution of the focal spot on this instrument in the past ten years.

Kinetics of oil saponification by lead salts in ancient preparations of pharmaceutical lead plasters and painting lead mediums.

Lead soaps can be found in archaeological cosmetics as well as in oil paintings, as product of interactions of lead salts with oil. In this context, a better understanding of the formation of lead soaps allows a follow-up of the historical evolution of preparation recipes and provides new insights into conservation conditions. First, ancient recipes of both pharmaceutical lead plasters and painting lead mediums, mixtures of oil and lead salts, were reconstructed. The ester saponification by lead salts is determined by the preparation parameters which were quantified by FT-IR spectrometry. In particular, ATR/FT-IR spectrometer was calibrated by the standard addition method to quantitatively follow the kinetics of this reaction. The influence of different parameters such as temperature, presence of water and choice of lead salts was assessed: the saponification is clearly accelerated by water and heating. This analysis provides chemical explanations to the historical evolution of cosmetic and painting preparation recipes.

Aortic insufficiency in Reiter's syndrome of juvenile onset.

Reiter's syndrome is an uncommon cause of childhood arthritis and aortic incompetence is extremely rare in these patients. A 7-year-old boy with Reiter's syndrome and hemodynamically significant aortic insufficiency is reported. He was HLD-B27 positive and valvular incompetence was detected 32 months after the onset while active synovitis was still present.