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Background: Ursolic acid (UA) is a natural triterpenoid which possesses anti-cancer activity. However, little is known regarding the activity and molecular mechanism of UA in cholangiocarcinoma (CCA). Thus, we investigated the effects of UA on growth inhibition and apoptosis induction through biomolecular changes in KKU-213 and KKU-055 CCA cell lines., Methods: The anti-proliferative effect of UA against CCA cells was evaluated using SRB assay. Changes in biomolecules were assessed by SR-FTIR microspectroscopy combined with PCA and conventional methods (i.e., Annexin V-FITC/PI staining for lipid alteration and apoptosis induction; Western blot analysis and caspase-3/7 activity assay for apoptotic protein detection)., Results: UA suppressed the proliferation of CCA cells in a dose- and time-dependent manner. SR-FTIR data revealed a significant alteration in lipids attributable to changes in apoptotic cell membranes, confirmed by Annexin V-FITC/PI staining. SR-FTIR data showed that UA promoted changes in the protein secondary structure. Elevated expression of Bax and decreased expression of Bcl-2 and survivin/BIRC5 along with augmented caspase-3/7 activity supported alterations in apoptosis-related proteins., Conclusions: SR-FTIR microspectroscopy was successfully used as a label-free technique to monitor apoptosis-induced biomolecular changes in UA-treated CCA cells. UA exerted the cytotoxic and apoptotic activities in CCA cells through alterations in membrane lipids and apoptotic proteins. UA could be a potential anti-CCA candidate and a chemical starting point for the discovery of novel anti-cancer agents., Significance: Our present study showed the first evidence that UA exhibited the anti-proliferative and pro-apoptotic activities toward CCA cells through changes in biomolecules, notably lipids and proteins., Competing Interests: Declaration of Competing Interest The authors disclose no conflicts of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The objective of this study was to use advanced vibrational molecular spectroscopy (ATR-Ft/IRS) to study an interaction between legume protein molecular structure from biodegradation residues and nutritional properties of newly developed genotypes of cool-climate adapted faba bean seed with low and normal tannin levels grown in western Canada. Protein molecular structures including amide I, II areas and peak heights, α-helix and β-sheet peak heights in rumen biodegradation residues were determined by using attenuated total reflectance Fourier transform infrared molecular spectroscopy (ATR-FTIR). The nutritional properties were determined which included chemical and nutrient profiles, in situ rumen degradation kinetics, rumen protein degradation, and intestinal protein digestion in the newly developed genotypes of faba bean seeds with low and normal tannin levels. The results showed that the spectral intensity of faba bean varieties with a normal tannin level in rumen biodegradation residues was greater (P < 0.05) than that with a low tannin level. The spectral intensity of amide I, II areas and peak heights, α-helix and β-sheet peak heights in all genotypes (except the variety of Snowdrop) in biodegradation residuals of faba bean seeds had a unique pattern with increasing first and then decreasing with the increasing of rumen incubation time. The molecular structures of protein (α-helix, ratio of α-helix to β-sheet height and amide I to II area, R 2  > 0.6) were associated with in situ degradation kinetics - soluble (S) and potential degradable fractions (D) and rumen protein degradation- bypass or undegraded protein (BCP or RUP). The molecular spectral parameters in the FTIR fingerprint region didn't form cluster among different genotypes in residual faba bean seeds in 12 h and 24 h incubation, which indicate they had similar protein molecular structures after incubation. In conclusion, there was an interaction between protein molecular structure from biodegradation residues and nutritional properties of newly developed cool-climate adapted faba bean seeds with normal and low level of tannin. The cool-climate adapted genotype had an impact on the protein molecular structure, and the protein utilization and metabolism were predictable from protein spectral molecular structures after rumen biodegradation with ATR-Ft/IRS spectroscopy., Competing Interests: Declaration of Competing Interest We state that there are no conflicts of interest., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Since the discovery of X-ray diffraction and its potential to elucidate crystal symmetry, powder X-ray diffraction has found diverse applications in the field of pharmaceutical sciences. This review summarizes significant achievements of the technique during various stages of dosage form development. Improved understanding of the principle involved and development of automated hardware and reliable software have led to increased instrumental sensitivity and improved data analysis. These advances continue to expand the applications of powder X-ray diffraction to emerging research fields such as amorphous systems, mechanistic understanding of phase transformations, and "Quality by Design" in formulation development., (Copyright © 2018 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.)

Following pioneering work 40 years ago, synchrotron beamlines dedicated to macromolecular crystallography (MX) have improved in almost every aspect as instrumentation has evolved. Beam sizes and crystal dimensions are now on the single micron scale while data can be collected from proteins with molecular weights over 10 MDa and from crystals with unit cell dimensions over 1000 Å. Furthermore it is possible to collect a complete data set in seconds, and obtain the resulting structure in minutes. The impact of MX synchrotron beamlines and their evolution is reflected in their scientific output, and MX is now the method of choice for a variety of aims from ligand binding to structure determination of membrane proteins, viruses and ribosomes, resulting in a much deeper understanding of the machinery of life. A main driving force of beamline evolution have been advances in almost every aspect of the instrumentation comprising a synchrotron beamline. In this review we aim to provide an overview of the current status of instrumentation at modern MX experiments. The most critical optical components are discussed, as are aspects of endstation design, sample delivery, visualisation and positioning, the sample environment, beam shaping, detectors and data acquisition and processing., (Copyright © 2016. Published by Elsevier Inc.)

A small angle X-ray scattering (SAXS) synchrotron was used to evaluate 100 ovine m. longissimus lumborum, representing lamb (n=50) and sheep (n=50). The diffraction of X-rays gives information on muscle myofibril structure and fat content. The linear relationships between SAXS measures with measures such as, shear force, intramuscular fat content (IMF) and collagen content/solubility, were investigated. A relationship was found between the d-spacing of the actin/myosin fibril spacing (SAX1 and SAX2) and the cross sectional area of the rhombohedral unit cell (Cell area) and shear force after 1 and 5day ageing. There was a positive relationship between IMF and a SAXS Fat area measure. There was a muscle site effect on SAX1, SAX2 and Cell area, with the cranial site having a larger distance between myofibrils. The potential of SAXS as a powerful research tool to determine not only the structural components of ovine tenderness, but also the fat content related to IMF is evident., (Crown Copyright © 2016. Published by Elsevier Ltd. All rights reserved.)

Introduction: This study evaluated the integrity of calcium silicate sealer-based fillings made with hygro-expandable cones (HEC) that are commercially known as CPoint or Smartpoint., Methods: Fourteen human canines were prepared according to a standardized, conventional endodontic treatment protocol and filled with the HEC/calcium silicate sealer. Three-dimensional imaging was performed with laboratory micro-computed tomography (μCT) at its highest resolution and was compared with synchrotron phase contrast-enhanced μCT (PCE-CT) scans of the treatment extending 1-7 mm from the apex. Conventional destructive optical microscopy validated observations by comparison with virtual slices in the tomographic data., Results: Conventional laboratory μCT at 10-μm resolution did not reveal the existing voids and defects within the root canal fillings. PCE-CT revealed elongated interfacial delamination localized mainly at the HEC-sealer interface forming extended through-and-through gaps along the root canal filling., Conclusions: Endodontic studies that use conventional laboratory μCT may underestimate thin defects and delamination within root canal fillings made with HEC because of limited resolution and contrast of laboratory-based broad-spectrum low intensity x-ray sources. These limitations favor use of high-brilliance, monochromatic synchrotron-based PCE-CT to reveal the important micrometer details within large (millimeter sized) samples. PCE-CT revealed the existence of a range of significant structural defects in recently placed HEC fillings, confirmed by optical microscopy after physical sectioning. Substantial delamination spanning 20%-40% of the circumferential interface as well as other structural defects were identified within root canal fillings made of HEC and calcium silicate sealer., (Copyright © 2016 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Human exposure to asbestos can cause a wide variety of lung diseases that are still a current major health concern, even if asbestos has been banned in many countries. It has been shown in many studies that asbestos fibers, ingested by alveolar macrophages, disrupt lung iron homeostasis by sequestering iron. Calcium can also be deposited on the fibers. The pathways along which iron and above all calcium interact with fibers are still unknown. Our aim was that of investigating if the iron accumulation induced by the inhaled asbestos fibers also involves calcium ions accumulation. Lung sections of asbestos-exposed mice were analyzed using an extremely sensitive procedure available at the synchrotron facilities, that provides morphological and chemical information based on X-ray fluorescence microspectroscopy (μ-XRF). In this study we show that (1) where conventional histochemical procedures revealed only weak deposits of iron and calcium, μ-XRF analysis is able to detect significant deposits of both iron and calcium on the inhaled asbestos fibers; (2) the extent of the deposition of these ions is proportionally directly related and (3) iron and calcium deposition on inhaled asbestos fibers is concomitant with the appearance of inflammatory and hyperplastic reactions., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)

There have been increasing interests in studying biological effects of nanomaterials, which are nevertheless faced up with many challenges due to the nanoscale dimensions and unique chemical properties of nanomaterials. Synchrotron-based X-ray microscopy, an advanced imaging technology with high spatial resolution and excellent elemental specificity, provides a new platform for studying interactions between nanomaterials and living systems. In this article, we review the recent progress of X-ray microscopic studies on bioeffects of nanomaterials in several living systems including cells, model organisms, animals and plants. We aim to provide an overview of the state of the art, and the advantages of using synchrotron-based X-ray microscopy for characterizing in vitro and in vivo behaviors and biodistribution of nanomaterials. We also expect that the use of a combination of new synchrotron techniques should offer unprecedented opportunities for better understanding complex interactions at the nano-biological interface and accounting for unique bioeffects of nanomaterials., From the Clinical Editor: Synchrotron-based X-ray microscopy is a non-destructive imaging technique that enables high resolution spatial mapping of metals with elemental level detection methods. This review summarizes the current use and perspectives of this novel technique in studying the biology and tissue interactions of nanomaterials., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Profiles of the major sulfur functional groups in mature Norway spruce wood tissue have been established for the first time. The big challenge was the development of a method suitable for sulfur speciation in samples with very low sulfur content (<100 ppm). This became possible by synchrotron X-ray absorption spectroscopy at the sulfur L-edge in total electron yield (TEY) detection mode with thin gold-coated wood slices. Functional groups were identified using sulfur compound spectra as fingerprints. Latewood of single year rings revealed metabolic plausible sulfur forms, particularly inorganic sulfide, organic disulfide, methylthiol, and highly oxidized sulfur. Form-specific profiles with Norway spruces from three different Swiss forest sites revealed high, but hitherto little-noticed, sulfur intensities attributable to natural heartwood formation and a common, but physiologically unexpected maximum around year ring 1986 with trees from the industrialized Swiss Plateau. It is hypothesized whether it may have resulted from the huge reduction in sulfur emissions after 1980 due to Swiss policy. Comparison with total S content profiles from optical emission spectroscopy underlined the more accurate and temporally better resolved TEY data with single wood year rings and it opened novel insights into the wood cell chemistry.

Radiolysis of water by synchrotron X-rays generates oxygen-containing radicals that undergo reactions with solvent accessible sites of macromolecules inducing stable covalent modifications or cleavage on millisecond time scales. The extent and site of these reactions are determined by gel electrophoresis and mass spectrometry analysis. These data are used to construct a high-resolution map of solvent accessibility at individual reactive sites. The experiments can be performed in a time-resolved manner to provide kinetic rate constants for dynamic events occurring at individual sites within macromolecules or can provide equilibrium parameters of binding and thermodynamics of folding processes. The application of this synchrotron radiolysis technique to the study of lysozyme protein structure and the equilibrium urea induced unfolding of apomyoglobin are described. The Mg2+-induced folding of Tetrahymena thermophila group I ribozyme shows the capability of the method to study kinetics of folding., (Copyright 2001 Academic Press.)

Nitric oxide (NO) produced by endothelial NO synthase (eNOS) plays a central role in regulation of vascular tone and reactivity. The purpose of this study is to clarify the basal tone and microvascular reactivity in eNOS-overexpressing transgenic (Tg) mice in vivo with a microangiography system using monochromatic synchrotron radiation (SR). The mouse femoral artery was cannulated, nonionic contrast media was injected, and microangiography was performed in hindlimbs of mice. Serial images of the small blood vessels (diameter <200 microm) were recorded by the SR microangiography system. At basal conditions, the diameter of tibial arteries in eNOS-Tg mice was larger than that of wild-type mice (179 +/- 8 versus 132 +/- 8 microm; P < 0.01). l-NAME treatment decreased the vessel diameter and canceled the difference in vessel diameters between two genotypes. Acetylcholine- and sodium nitroprusside-induced relaxations of small vessels were significantly reduced in Tg mice compared with wild-type mice (35.0 +/- 9.4 versus 61.6 +/- 6.7%, 85.0 +/- 10.2 versus 97.3 +/- 6.7% of the maximum relaxation, respectively). Our data provide the evidence that overproduced NO from endothelium reduces vascular tone and plays a pivotal role in regulation of vascular tone in small vessels. Furthermore, the reduced NO-mediated relaxation in small vessels of eNOS-Tg mice is demonstrated for the first time in vivo. SR microangiography allows us to evaluate the reactivity in small-sized vessels and appears to be a powerful tool for assessing the microvascular circulation in vivo., (Copyright 2001 Academic Press.)