Heat capacity and entropy behavior of andradite: a multi-sample and −methodological investigation

Andradite, ideal end-member formula Ca_3 Fe^(3+)_2Si_3O_(12), is one of the common rock-forming garnets found in the Earth's crust. There are several outstanding questions regarding andradite's thermodynamic and physical property behavior. Three issues are: i) Could there be differences in the thermodynamic properties, namely heat capacity, C_p , between synthetic and natural andradite crystals, as observed in the Ca-garnet grossular, Ca_3Al_2Si_3O_(12)? ii) What is the thermal nature of the low-temperature magnetic-phasetransition behavior of andradite? and iii) How quantitative are older published calorimetric (i. e., adiabatic and DSC) heat-capacity results? In this work, four natural nearly end-member single crystals and two synthetic polycrystalline andradite samples were carefully characterized by optical microscope examination, X-ray powder diffraction, microprobe analysis, and IR and UV/VIS single-crystal spectroscopy. The IR spectra of the different samples commonly show a main intense OH stretching band located at 3563 cm^(-1), but other OH bands can sometimes be observed as well. Structural OH concentrations, calculated from the IR spectra, vary from about 0.006 to 0.240 wt% H_2O. The UV/VIS spectra indicate that there can be slight, but not fully understood, differences in the electronic state between synthetic and natural andradite crystals. The C_p behavior was determined by relaxation calorimetry between 2 and 300 K and by differential scanning calorimetry (DSC) methods between 150/300 and 700/950 K, employing the same andradite samples that were used for the other characterization measurements. The low-temperature C_p results show a magnetic phase transition with a Néel temperature of 11.3 ± 0.2 K, which could be slightly affected by the precise electronic state of Fe^(2+/3+) in the crystals. The published adiabatic calorimetry results on andradite do not provide a full and correct thermal description of this magnetic transition. The calorimetric Cp measurements give a best estimate for the standard third-law entropy at 298.15 K for andradite of S^o ≈ 324 ± 2 J/mol · K vs. the value of 316.4 ± 2.0 J/mol · K, as given in an early adiabatic investigation. Both natural and synthetic crystals give similar S o values within experimental uncertainty of about 1.0%, but one natural andradite, richer in OH, may have a very slightly higher value around S^o≈ 326 J/mol·K. Low-temperature DSC measurements made below 298 K agree excellently with those from relaxation calorimetry. The DSC measurements above 298 K show a similarity in C_p behavior among natural and synthetic andradites. A C_p polynomial for use above room temperature to approximately 1000 K was calculated from the data on synthetic andradite giving: C_p (J/mol·K) = 599.09 (±14) 2709.5 (±480)· T^(0.5) 1.3866 (±0.26)· 10^7 · T^2 + 1.6052 (±0.42) · 10^9 · T^3

Murine Metatarsus Bone and Joint Collagen-I Fiber Morphologies and Networks Studied With SHG Multiphoton Imaging

Chronic inflammatory disease of bones and joints (e.g., rheumatoid arthritis, gout, etc.), but also acute bone injury and healing, or degenerative resorptive processes inducing osteoporosis, are associated with structural remodeling that ultimately have impact on function. For instance, bone stability is predominantly orchestrated by the structural arrangement of extracellular matrix fibrillar networks, i.e., collagen-I, -IV, elastin, and other proteins. These components may undergo distinct network density and orientation alterations that may be causative for decreased toughness, resilience and load bearing capacity or even increased brittleness. Diagnostic approaches are usually confined to coarse imaging modalities of X-ray or computer tomography that only provide limited optical resolution and lack specificity to visualize the fibrillary collagen network. However, studying collagen structure at the microscopic scale is of considerable interest to understand the mechanisms of tissue pathologies. Multiphoton Second Harmonic Generation (SHG) microscopy, is able to visualize the sterical topology of the collagen-I fibrillar network in 3D, in a minimally invasive and label-free manner. Penetration depths exceed those of conventional visible light imaging and can be further optimized through employing decalcification or optical clearing processing ex vivo. The goal of this proof-of-concept study was to use SHG and two-photon excited fluorescence (2-PEF) imaging to mainly characterize the fibrillary collagen organization within ex vivo decalcified normal mouse metatarsus bone and joint. The results show that the technique resolved the fibrillar collagen network of complete bones and joints with almost no artifacts and enabled to study the complex collagen-I networks with various fiber types (straight, crimped) and network arrangements of mature and woven bone with high degree of detail. Our imaging approach enabled to identify cavities within both cortical and trabecular bone architecture as well as interfaces with sharply changing fiber morphology and network structure both within bone, in tendon and ligament and within joint areas. These possibilities are highly advantageous since the technology can easily be applied to animal models, e.g., of rheumatoid arthritis to study structural effects of chronic joint inflammation, and to many others and to compare to the structure of human bone

Heat capacity and entropy behavior of andradite: a multi-sample and −methodological investigation

Andradite, ideal end-member formula Ca_3 Fe^(3+)_2Si_3O_(12), is one of the common rock-forming garnets found in the Earth's crust. There are several outstanding questions regarding andradite's thermodynamic and physical property behavior. Three issues are: i) Could there be differences in the thermodynamic properties, namely heat capacity, C_p , between synthetic and natural andradite crystals, as observed in the Ca-garnet grossular, Ca_3Al_2Si_3O_(12)? ii) What is the thermal nature of the low-temperature magnetic-phasetransition behavior of andradite? and iii) How quantitative are older published calorimetric (i. e., adiabatic and DSC) heat-capacity results? In this work, four natural nearly end-member single crystals and two synthetic polycrystalline andradite samples were carefully characterized by optical microscope examination, X-ray powder diffraction, microprobe analysis, and IR and UV/VIS single-crystal spectroscopy. The IR spectra of the different samples commonly show a main intense OH stretching band located at 3563 cm^(-1), but other OH bands can sometimes be observed as well. Structural OH concentrations, calculated from the IR spectra, vary from about 0.006 to 0.240 wt% H_2O. The UV/VIS spectra indicate that there can be slight, but not fully understood, differences in the electronic state between synthetic and natural andradite crystals. The C_p behavior was determined by relaxation calorimetry between 2 and 300 K and by differential scanning calorimetry (DSC) methods between 150/300 and 700/950 K, employing the same andradite samples that were used for the other characterization measurements. The low-temperature C_p results show a magnetic phase transition with a Néel temperature of 11.3 ± 0.2 K, which could be slightly affected by the precise electronic state of Fe^(2+/3+) in the crystals. The published adiabatic calorimetry results on andradite do not provide a full and correct thermal description of this magnetic transition. The calorimetric Cp measurements give a best estimate for the standard third-law entropy at 298.15 K for andradite of S^o ≈ 324 ± 2 J/mol · K vs. the value of 316.4 ± 2.0 J/mol · K, as given in an early adiabatic investigation. Both natural and synthetic crystals give similar S o values within experimental uncertainty of about 1.0%, but one natural andradite, richer in OH, may have a very slightly higher value around S^o≈ 326 J/mol·K. Low-temperature DSC measurements made below 298 K agree excellently with those from relaxation calorimetry. The DSC measurements above 298 K show a similarity in C_p behavior among natural and synthetic andradites. A C_p polynomial for use above room temperature to approximately 1000 K was calculated from the data on synthetic andradite giving: C_p (J/mol·K) = 599.09 (±14) 2709.5 (±480)· T^(0.5) 1.3866 (±0.26)· 10^7 · T^2 + 1.6052 (±0.42) · 10^9 · T^3

A Novel Bone Substitute with High Bioactivity, Strength, and Porosity for Repairing Large and Load-Bearing Bone Defects.

Achieving adequate healing in large or load-bearing bone defects is highly challenging even with surgical intervention. The clinical standard of repairing bone defects using autografts or allografts has many drawbacks. A bioactive ceramic scaffold, strontium-hardystonite-gahnite or "Sr-HT-Gahnite" (a multi-component, calcium silicate-based ceramic) is developed, which when 3D-printed combines high strength with outstanding bone regeneration ability. In this study, the performance of purely synthetic, 3D-printed Sr-HT-Gahnite scaffolds is assessed in repairing large and load-bearing bone defects. The scaffolds are implanted into critical-sized segmental defects in sheep tibia for 3 and 12 months, with bone autografts used for comparison. The scaffolds induce substantial bone formation and defect bridging after 12 months, as indicated by X-ray, micro-computed tomography, and histological and biomechanical analyses. Detailed analysis of the bone-scaffold interface using focused ion beam scanning electron microscopy and multiphoton microscopy shows scaffold degradation and maturation of the newly formed bone. In silico modeling of strain energy distribution in the scaffolds reveal the importance of surgical fixation and mechanical loading on long-term bone regeneration. The clinical application of 3D-printed Sr-HT-Gahnite scaffolds as a synthetic bone substitute can potentially improve the repair of challenging bone defects and overcome the limitations of bone graft transplantation

Measurement of friction-induced changes in pig aorta fibre organization by non-invasive imaging as a model for detecting the tissue response to endovascular catheters

Alterations in quantity or architecture of elastin and collagen fibres are associated with some blood vessel pathologies. Also some medical interventions such as endovascular catheterization have the potential to damage blood vessels. This study reports the use of porcine aorta as a model system for studying the physical impact of catheters on vasculature, in conjunction with non-invasive imaging techniques to analyse collagen and elastin fibre organization and assess load-induced changes. Porcine aorta was exposed to frictional trauma and elastin and collagen fibre orientation evaluated by destructive, histochemical methods and non-invasive imaging. The latter allowed the immediate impact of force on fibre orientation and fibre recovery to be evaluated longitudinally. In normal aorta, elastin fibres are aligned at the surface, but become less aligned with increasing depth, showing no alignment by ~30 µm. Collagen fibres meanwhile appear aligned down to a depth of 35 µm. Changes in collagen and elastin fibre orientation in healthy pig aorta were detected by conventional destructive histology within 5 minutes of application of a sliding 10N load, while lesser loads had less impact. Good recovery of fibre orientation was observed within 20 minutes. Non-invasive imaging of ex vivo aorta tissue provides a good indication of the extent of fibre re-organization following frictional stress, at loads similar to those encountered during medical interventions such as catheterization. These results indicate that tissue deformation can occur from these procedures, even in healthy tissue, and highlight the potential for the development of an in vivo probe capable of monitoring vascular changes in patients

Ore-forming processes of the daqiao epizonal orogenic gold deposit, west qinling orogen, China: Constraints from textures, trace elements, and sulfur isotopes of pyrite and marcasite, and raman spectroscopy of carbonaceous material

The Daqiao gold deposit is hosted in organic-rich Triassic pumpellyite-actinolite facies metamorphosed turbidites in the West Qinling orogen, central China. Gold mineralization is characterized by high-grade hydraulic breccias (B and C ores) that overprint an earlier tectonic breccia (A ore). A complex paragenesis is defined by four sulfide stages: S1 diagenetic preore pyrite (py), S2 hydrothermal early ore disseminated pyrite and marcasite (mc), S3 main ore pyrite and marcasite aggregates, and S4 late ore coarse-grained marcasite with minor pyrite and stibnite. However, multiple generations of pyrite and marcasite may develop within one individual stage. Ore-related hydrothermal alteration is dominated by intensive silicification, sulfidation, sericitization, and generally distal minor carbonatization. Laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) trace element analyses show that the stage S1 py1 from the shale interlayers within turbidites contains low gold contents (mean of 0.05 ppm) and other trace elements (Mn, Co, Ni, Cu, Mo, Bi, and Pb), indicating an anoxic to euxinic sedimentary environment. Stage S2 contributed only minimally to the gold endowment with relatively low gold in various sulfides including py2 (mean of 0.09 ppm), py3 (0.84 ppm) to py4 (0.70 ppm), along with mc1 (0.02 ppm) and mc2 (0.14 ppm). Most of the gold was deposited in stage S3, which formed rapidly crystallized, irregular (e.g., framboids, colloform and cyclic zonation) cement-hosted py5a (mean of 27.35 ppm), py5b (9.71 ppm), and mc3 (5.94 ppm) during repeated hydraulic fracturing. Other trace elements (e.g., Ag, As, Sb, Hg, Tl, and W) are also significantly enriched in the main ore-stage pyrite and marcasite. Little or no gold is detected in the S4 py6 and mc4. Sulfur isotopes determined from in situ LA-multicollector (MC)-ICP-MS analyses of hydrothermal pyrite and marcasite from the Daqiao deposit vary significantly from –31.3 to 22.0 (d34S values) but fall mostly between –10 to 10 and provide important information on the source and evolution of sulfur and of the ore-forming fluids. The results show that S2 ore fluids (mean d34Ssulfide = –0.8 to 5.2) were most likely derived from deep-seated Paleozoic carbonaceous sediments during regional metamorphism associated with orogenesis of the West Qinling orogen. Main ore S3 fluids (mean d34Ssulfide = –9.7 to –6.0) are relatively depleted in34S relative to those of S2, presumably due to fluid oxidation associated with hydraulic fracturing caused by the overpressurized fluids. The textural, chemical, and isotopic data indicate two distinct gold-introducing episodes at Daqiao, forming sulfide disseminations during early ore S2 and cement-hosted sulfide aggregates during main ore S3. The S2 mineralization took place in a tectonic breccia beneath low-permeability shale seals that capped the flow of deep-seated metamorphic fluids, facilitating reaction with preexisting carbonaceous material and the host turbidites to form sulfide disseminations and pervasive silicification. Raman spectroscopy analysis suggests that carbonaceous material in the ores is poorly crystallized, with low maturity, giving estimated temperatures of 283° to 355°C that are much higher than those of the ore fluids (100°–240°C). This temperature difference indicates an in situ sedimentary origin modified by the regional pumpellyite-actinolite facies metamorphism for the carbonaceous material in the host rocks, rather than a hydrothermal origin. In S3, continuous flux of hydrothermal fluids caused fluid overpressure and consequent hydraulic fracturing of the competent silicified rocks. Subsequent rapid fluid pressure fluctuations led to phase separation and thus massive oxidation of ore fluids, which triggered fast precipitation of gold and other trace elements within the fine-grained irregular sulfides. Results presented here, in combination with geologic evidences, suggest that the Daqiao gold deposit can be best classified as the shallow-crustal epizonal orogenic type, genetically associated with orogenic deformation and regional metamorphism of the West Qinling orogen

Studies on the inactivation process of the tyrosine kinase Src in the integrin alphaIIb-beta3 signaling pathway by fluorescence microscopy

Essentiell für die Blutstillung (Haemostase) ist die Thrombozyten- oder Blutplaettchen-Adhaesion und die Thrombus-Bildung. Beide Vorgaenge werden hauptsaechlich durch den Thrombozyten-Rezeptor Integrin alphaIIb-beta3 vermittelt. Nach Bindung des Liganden Fibrinogen aendert sich die Rezeptor-Konformation, Integrine assoziieren und ein intrazellulaeres Signalnetzwerk wird aktiviert, welches die Organisation des Aktin-Zytoskeletts steuert. Diese Zytoskelett-Reorganisationen sind Grundlage für zellulaere Adhaesions- und Aggregations-Prozesse. Die Signalvermittlung vom Integrin zum Zytoskelett wird durch die Protein-Tyrosinkinase Src eingeleitet, deren Aktivitaetszustand den Signalweg reguliert. Bei der Src-Aktivierung wird Tyrosin 418 durch Autokatalyse phosphoryliert. Die Kinase muss jedoch wieder inaktiviert werden. Dies übernimmt in Plaettchen ausschliesslich die Tyrosinkinase Csk (C-terminale Src Kinase) durch Phosphorylierung von Tyrosin 529 im C-terminalen Ende des Proteins. Die Csk-vermittelte Inaktivierung von Src stellt den entscheidenden Kontrollschritt des alphaIIb-beta3-vermittelten Signalwegs dar. Obwohl bekannt ist, dass die Src-Aktivierung bei der Zelladhaesion an den Zellraendern der Lamellipodien geschieht und man den Mechanismus und die Kinetik der Src-Csk Interaktion genauer versteht, ist bislang immer noch unbekannt, wo und wie Src inaktiviert wird bzw. welche Rolle der Src-Inaktivierung genau zukommt. FRET (Fluoreszenz-Resonanz-Energie-Transfer) ist ein physikalischer Effekt, mit dem Interaktionen beliebiger fluoreszenzmarkierter Proteine mikroskopisch detektiert werden koennen. Diese Technik wurde genutzt, um die Src-Csk-Interaktion waehrend der alphaIIb-beta3-vermittelten Fibrinogen-Adhaesion in einer etablierten Thrombozyten-Modellzelllinie (A5-CHO) direkt visualisierbar zu machen. Es zeigten sich starke Src-Csk Interaktionen (FRET-Signale) an den Zellraendern aktiver Lamellipodien und zusaetzlich in Fokalkontakten, wo beide Proteine mit Vinculin, einem Fokalkontakte-Marker, co-lokalisierten. Die Proteininteraktionen folgten einem hochdynamischen Ablauf. Nach der Akkumulation der Src-Csk Komplexe an den Zellraendern wanderten sie in Abstaenden von 2-3 Minuten nach innen, fragmentierten und bildeten schliesslich stabile Fokal-Adhaesionen. FRET-Signale an den Zellraendern fanden sich vor allem in ruhenden Lamellipodien bzw., waehrend des Lamellipodien-Rückzugs, in wachsenden Lamellipodien traten die FRET-Signale dort dagegen nicht auf. In unabhaengigen biochemischen Tests im Zeitfenster der FRET-Beobachtungen wurde ein spezifischer Anstieg der Src-Tyr529-Phosphorylierung (Inaktivierung) und eine parallele Abnahme der Src-Tyr418-Phosphorylierung (Aktivierung) gemessen. Weiterführende Ergebnisse lieferten Versuche mit Src- und Csk-Mutanten. Die Co-Expression von Wildtyp-Src mit Kinase-inaktivem CskK222R hatte weder einen Effekt auf die Adhaesion und Ausbreitung der Zellen noch auf die Praesenz von FRET, es aenderte sich jedoch drastisch die zellulaere Verteilung der FRET-Signale sowie das Wachstum und die Form der Lamellipodien. Die Co-Expression von Wildtyp-Csk mit konstitutiv aktivem SrcY529F verursachte dagegen eine stark verringerte Adhaesionsfaehigkeit und Hemmung der Lamellipodien-Bildung. Die Fokal-Adhaesionspunkte in diesen Zellen waren sehr schwach und ueberdimensioniert und lagen ungeordnet verteilt in der Adhaesionsebene. Zusaetzlich verursachte SrcY529F eine starke Ueberaktivierung des Zytoskeletts und das fast vollstaendige Verschwinden der FRET-Signale. Die ermittelten Daten zeigen, dass die enge Kontrolle der Src-Aktivitaet durch Csk eine bedeutende Rolle für die funktionelle Zell-Adhaesion and -Ausbreitung spielt. Co-Immunpraezipitations-Resultate und Messungen der Menge an markiertem Protein in Zellen, in welchen FRET detektierbar war, untermauern zusaetzlich unsere These, zum ersten Mal die Src-Regulation durch Csk in lebenden Zellen direkt beobachtbar gemacht zu haben. Dieser neue FRET-Ansatz kann auch als Reporter-System für Prozesse der Src-Inaktivierung in anderen Signalwegen und Zellen angewendet werden. Das Messprinzip kann weiterhin auf das Studium der Inaktivierung weiterer Mitglieder der Familie der Src-Kinasen (in verschiedensten Signalwegen) erweitert werden.Platelet adhesion and thrombus formation required for functional hemostasis depends on integrin receptor mediated “outside-in” signaling to the cytoskeleton. Integrin alphaIIb-beta3 is the major integrin on the platelet surface and acts as a specific receptor for the plasma protein fibrinogen. Fibrinogen binding causes clustering of integrins within the plasma membrane activating the protein tyrosine kinase Src (signal initiation) by phosphorylation of tyrosine 418. Src, however, is negatively regulated by another tyrosine kinase, Csk (C-terminal Src kinase), which phosphorylates tyrosine 529. Although, in adhering cells, it is believed that Src is getting activated at lamellipodia leading edges, neither the cellular location nor the dynamics and exact role of Src inactivation is known to date. Here, we studied Src inactivation during alphaIIb-beta3-dependent adhesion to fibrinogen in the established platelet model cell line A5-CHO. Using a live cell FRET (fluorescence resonance energy transfer) microscopy technique with CFP and YFP label molecules (cyan and yellow fluorescent protein), we were able to image highly dynamic Src-Csk interactions at the leading edges of active lamellipodia. Every 2-3 minutes, signals detecting Src-Csk interactions (complexes) appeared at the cell periphery before they begin to move inward in the cell and reorganize while lamellipodia start to protrude (grow). FRET signals were also found in small accumulations at the fringe and also further to the centre of the adhesion plane (focal complexes and adhesions). Src and Csk co-localize with vinculin (a focal adhesion marker) within these regions. During the runtime of FRET observation a specific increase in Src-Tyr529 phosphorylation with a parallel decrease in Src-Tyr418 phosphorylation was observed supporting the idea that Src inactivation occurs within the cells. The role of Src-Csk interaction was studied in further detail using Src and Csk mutants. The data revealed that co-expression of inactive CskK222R did not alter the presence of FRET signals, but fundamentally changed its distribution within the cell. Furthermore it caused lamellipodia shape changes and a tendency of constant lamellipodia protrusion. Co-expression of constitutively active SrcY529F in turn caused a severe adhesion and spreading dysfunction. Adherent cells showed very weak, disorganized and oversized focal adhesions, a hyper-activated cytoskeleton (visible in fast-changing membrane blebs) and absence of FRET signals. Results from immunoprecipitation analyses and protein level determination within cells, in which FRET was detectable, further supported that we were able, for the first time, to directly visualize Src (and integrin) regulation by Csk control in live cells. The results show that Src control by Csk is ultimately required for lamellipodia and focal adhesion function and thus for cell anchorage and spreading. The novel FRET-approach reported here can be readily applied to other integrin and signaling pathways including the study of closely related Src family kinases (SFKs). Results may also contribute to a better understanding of the processes of tumor formation

The giant Kalgoorlie Gold Field revisited

© 2015 China University of Geosciences (Beijing) and Peking University. The Neoarchaean Kalgoorlie Gold Field contains the giant Golden Mile and world-class Mt Charlotte deposits, which have been the subject of much research for over 100 years. The Golden Mile deposit is a complex array of ductile to brittle vein and breccia lodes that are predominantly hosted in the highly-fractionated Golden Mile Dolerite sill. The Fimiston lodes comprise an array of narrow lodes that evolved broadly syn- to late-formation of the regional D2 NW-trending foliation. The lodes are characterized by pyrite veinlets and disseminations, quartz veinlets and breccias, and banded quartz-carbonate veins with sericite, carbonate, and pyrite-dominated alteration. Bonanza Green-Leader, or Oroya-style, lodes, with grades in excess of 1000 g/t Au, are similar to the Fimiston-style lodes, but are characterized by abundant visible gold, native tellurium and more abundant telluride minerals within roscoelite-bearing alteration zones. The arguably structurally younger Mt Charlotte-style lodes are characterized by a pipe-shaped, coarse-grained quartz, carbonate and scheelite vein-stockwork with distinct vertically-zoned, carbonate-sericite-albite-pyrite ± pyrrhotite dominant alteration assemblages around veins within Unit 8 of the Golden Mile dolerite and porphyry dykes. The network of steep- and gently-dipping extension and shear fracture-fill veins are associated with NE-trending fault sets that cross cut the regional NW-trend. The deposit area is intruded by swarms of porphyry dykes, including syn-volcanic mafic dykes, early and volumetrically most significant c. 2.67 Ga feldspar-phyric porphyry dykes, as well as later c. 2.66-2.65 Ga calc-alkaline hornblende-phyric dykes associated with younger c. 2.65-2.64 Ga lamprophyre dykes. All post-volcanic dykes have similar orientations to the Fimiston lodes. The feldspar dykes are clearly overprinted by all styles of mineralization, although the relationship between hornblende-phyric and lamprophyre dykes and gold mineralization is more ambiguous. Most agree that gold mineralization was post-peak regional metamorphism of host rocks, although its relative structural timing is controversial.Direct timing constraints on gold mineralization indicate that Fimiston- and Mt Charlotte-style mineralization formed within a relative short period of time around 2.64 Ga, and, as such, support a model of progressive deformation of a rheologically heterogeneous rock package late in the structural history. Fluid characteristics, combined with the structural, metamorphic and absolute timing, support description of gold mineralization at the Golden Mile as orogenic and mesozonal, and this allows direct correlation with orogenic gold deposits worldwide, which classically formed during accretion along convergent margins throughout Earth history