Secondary Cleavage Research Articles

Acetylene containing cyclo(L-Tyr-L-Tyr)-analogs as mechanism-based inhibitors of CYP121A1 from Mycobacterium tuberculosis

Tuberculosis (TB) is a globally significant infective disease that is caused by a single infectious agent, Mycobacterium tuberculosis (Mtb). Because of the rise in the number of multidrug-resistant (MDR) TB strains, identification of alternative drug targets for the development of drugs with different mechanism of actions is desired. CYP121A1, one of the twenty cytochrome P450 enzymes encoded in the Mtb genome, was previously shown to be essential for bacterial growth. This enzyme catalyzes the intramolecular C–C crosslinking reaction of the cyclopeptide cyclo(L-tyr-L-tyr) (cYY) yielding the metabolite mycocyclosin. In the present study, acetylene-substituted cYY-analogs were synthesized and evaluated as potential mechanism-based inhibitors of CYP121A1. The acetylene-substituted cYY-analogs were capable of binding to CYP121A1 with affinities comparable with cYY, and exhibited a Type I binding mode, indicative of a substrate-like binding, mandatory for metabolism. Only the cYY-analogs which contain an acetylene-substitution at one (2a) or both (3) para-positions of cYY showed mechanism-based inhibition of CYP121A1 activity. The values of KI and kinact were 236 µM and 0.045 min−1, respectively, for compound 2a, and 145 µM and 0.015 min−1, repectively, for compound 3 The inactivation could neither be reversed by dialysis nor be prevented by including glutathione. LC-MS analysis demonstrated that the inactivation results from covalent binding to the apoprotein, whereas the heme was unmodified. Interestingly, the mass increment of the CYP121A1 apoprotein was significantly smaller than was expected from the ketene formed by oxidation of the acetylene-group, indicative for a secondary cleavage reaction in the active site of CYP121A1. Although the two acetylene-containing cYY-analogs showed significant mechanism-based inhibition, growth inhibition of the Mtb strains was only observed at millimolar concentrations. This low efficacy may be due to insufficient irreversible inactivation of CYP121A1 and/or insufficient cellular uptake. Although the identified mechanism-based inhibitors have no perspective for Mtb-treatment, this study is the first proof-of-principle that mechanism-based inhibition of CYP121A1 is feasible and may provide the basis for new strategies in the design and development of compounds against this promising therapeutic target.

Numerical simulation on biomass-pyrolysis and thermal cracking of condensable volatile component

We studied the physical and chemical properties of the condensable volatiles of biomass pyrolysis products. We redefine the liquid product and divide the condensable volatiles into two categories, biomass oil and tar, the latter of which comes from the secondary pyrolysis or cracking reaction of the former. We further establish a kinetic model of biomass pyrolysis and secondary cracking. The chemical reaction kinetics equation and heat transfer equation are coupled to simulate the biomass pyrolysis process. For biomass solid particles, the model not only considers the initial reaction of biomass and secondary cleavage reaction of condensable gas, but also introduces a reaction mode in which biomass oil is converted into tar. When the pyrolysis temperature is below 500 °C, the pyrolysis products are essentially biomass oil. However, when the pyrolysis temperature exceeds 500 °C, the biomass oil gradually converts into tar. The model also considers characteristics of the reaction medium (porosity, intrinsic permeability, thermal conductivity) and the unsteady gas phase process based on Darcy's law of velocity and pressure, heat convection, diffusion, and radiation transfer. We analyze the relationships among the internal temperature of the particles, particle size and position, mass fraction of the reactants and products, the gas mixture, the production share of tar and biomass oil, and the relationship between gas pressure and time. The results show that the effects of the secondary cracking reaction and internal convective flow in the biomass pyrolysis process are coupled because the flow field in the porous medium determines the volatile residence time and thus species that affect the secondary cracking reaction. The rate of volatile formation in the initial and secondary cracking reactions affects the pressure gradient and gas diffusion. Additionally, the endothermic effect influences the temperature field of the pyrolysis reaction but has no apparent effect on small particles whose chemical reaction is the control mechanism. For large particles, heat transfer inside the particles is the diffusion control mechanism and the chemical reaction on the particle surface is the speed control mechanism. Two peaks are observed in the pyrolysis gas mass proportion curve, which result from the consumption of biomass oil and tar as they flow toward hot surfaces. The first peak is the decomposition of biomass oil into non-condensable volatile matter and tar, and the second peak is the further cracking of tar into gas and coke at high temperature.

Visualization of microstructural factors resisting the crack propagation in mesosegregated high-strength low-alloy steel

While relationship between fracture mechanism and homogeneous microstructures has been fully understood, relationship between fracture mechanism and inhomogeneous microstructures such as the mesosegregation receives less attention as it deserves. Fracture mechanism of the high-strength low-alloy (HSLA) steel considering the mesosegregation was investigated and its corresponding microstructure was characterized in this paper. Mesosegregation refers to the inhomogeneous distribution of alloy elements during casting solidification, and leads to the formation of positive segregation zones (PSZ) and negative segregation zones (NSZ) in ingots. The fracture surface of impact sample exhibits the quasi-cleavage fracture at −21 °C, and is divided into ductile and brittle fracture zone. Meanwhile, the PSZ and NSZ spread across ductile and brittle fracture zone randomly. In ductile fracture zone, micro-voids fracture mechanism covers the PSZ and NSZ, and higher deformation degree is shown in the PSZ. In brittle fracture zone, secondary cleavage cracks are observed in both PSZ and NSZ, but present bigger size and higher quantity in the NSZ. However, some regions of the PSZ still present micro-voids fracture mechanism in brittle fracture zone. It reveals that the microstructures in the PSZ exhibit a higher resistance ability to crack propagation than that in the NSZ. All observations above provide a better visualization of the microstructural factors that resist the crack propagation. It is important to map all information regarding the fracture mechanism and mesosegregation to allow for further acceptance and industrial use.

Efficient gene correction of an aberrant splice site in β-thalassaemia iPSCs by CRISPR/Cas9 and single-strand oligodeoxynucleotides.

β‐thalassaemia is a prevalent hereditary haematological disease caused by mutations in the human haemoglobin β (HBB) gene. Among them, the HBB IVS2‐654 (C > T) mutation, which is in the intron, creates an aberrant splicing site. Bone marrow transplantation for curing β‐thalassaemia is limited due to the lack of matched donors. The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR‐associated protein 9 (Cas9), as a widely used tool for gene editing, is able to target specific sequence and create double‐strand break (DSB), which can be combined with the single‐stranded oligodeoxynucleotide (ssODN) to correct mutations. In this study, according to two different strategies, the HBB IVS2‐654 mutation was seamlessly corrected in iPSCs by CRISPR/Cas9 system and ssODN. To reduce the occurrence of secondary cleavage, a more efficient strategy was adopted. The corrected iPSCs kept pluripotency and genome stability. Moreover, they could differentiate normally. Through CRISPR/Cas9 system and ssODN, our study provides improved strategies for gene correction of β‐Thalassaemia, and the expression of the HBB gene can be restored, which can be used for gene therapy in the future.

Uncovering the detailed mode of cleavage of heparinase I toward structurally defined heparin oligosaccharides

For a more insightful investigation into the specificity of bacterial heparinase I, a series of structurally well-defined heparin oligosaccharides was synthesized using a highly efficient chemoenzymatic strategy. Apart from the primary cleavage site, five glycosidic linkages of oligosaccharides with varying modifications to obtain secondary cleavage sites were degraded by a high concentration of heparinase I. The reactivity of linkages toward heparinase I was not entirely dependent on the 2-O-sulfated iduronic acid being cleaved or the neighboring 6-O-sulfated glucosamine residues, but it was dependent on higher degrees of sulfation of oligosaccharides and indispensable N-substituted glucosamine adjacent to the cleavable linkage. Moreover, the enzyme demonstrated less preferential cleavage toward glycosidic linkages containing glucuronic acid than those containing iduronic acid of the counterpart oligosaccharides. Biolayer interferometry revealed differences in reactivity that are not completely consistent with different affinities of substrates to enzyme. Our study presented accurate information on the cleavage promiscuity of heparinase I that is crucial for heparin depolymerization.

In silico analyses of molecular interactions between groundnut bud necrosis virus and its vector, Thrips palmi.

Groundnut bud necrosis virus (GBNV) is an economically important tospovirus transmitted by Thrips palmi (Thysanoptera: Thripidae). The current understanding of thrips-tospovirus interactions is largely based on the tomato spotted wilt virus-Frankliniella occidentalis relationship. Only limited information is available for the GBNV-T. palmi system. In the present study, available genome data of T. palmi and GBNV were used to predict the protein partners that may play a crucial role in the internalization of GBNV virions into thrips cells. Computational analyses showed that the GBNV precursor glycoprotein bears a signal peptide of 24 amino acids and a secondary cleavage site at position 434-435 separates the amino-terminal mature glycoprotein (GN) from the carboxyl-terminal glycoprotein (GC). Potential interactions of GBNV glycoproteins were predicted with T. palmi enolase, cathepsin, C-type lectin, clathrin and vacuolar ATP synthase subunit E. The in silico analyses suggested that C-type lectin is the primary cellular receptor to interact with GBNV-GN. After receptor binding, virus particles probably enter vector cells by clathrin-mediated endocytosis. This is the first in silico evidence of GBNV-T. palmi protein interaction.

Effect of hot deformation and crystallographic texture on toughness anisotropy and fracture behavior of Nb+V microalloyed API X70 steel

The Charpy impact toughness (CVN) properties of a Nb + V microalloyed X70 steel of YS∼ 540 MPa was studied at various angles with respect to the rolling direction. The microstructure of the steel was characterised using light optical microscope, scanning electron microscope (SEM) and the transformation textures were evaluated from the orientation distribution function (ODF) measured from the electron back-scattered diffraction (EBSD) experiments. The skeleton lines, showing intensity distribution, f(g), of the fcc-to-bcc transformation texture, namely the α-fiber (<110> ∥ RD), the γ-fiber (<111> ∥ ND) and ε-fiber (<110> ∥ TD) are considered to explain the observed behavior in mechanical properties. The major components like, {112}<110>, {113}<110>, {332}<113>, {111}<112> and {111}<110>, their relative intensity distribution, are found to act in a complex manner towards controlling the mechanical behavior during impact and tensile testing. It has been observed that, the type and extent of delamination normally observed in broken Charpy specimens, which is commonly believed to be more dependent on the banding of 2nd phases in the microstructure, is found to be controlled by crystallographic texture, mostly clustering of {100} grains in the primary and secondary cleavage planes.

Stabilized coronavirus spikes are resistant to conformational changes induced by receptor recognition or proteolysis

Severe acute respiratory syndrome coronavirus (SARS-CoV) emerged in 2002 as a highly transmissible pathogenic human betacoronavirus. The viral spike glycoprotein (S) utilizes angiotensin-converting enzyme 2 (ACE2) as a host protein receptor and mediates fusion of the viral and host membranes, making S essential to viral entry into host cells and host species tropism. As SARS-CoV enters host cells, the viral S is believed to undergo a number of conformational transitions as it is cleaved by host proteases and binds to host receptors. We recently developed stabilizing mutations for coronavirus spikes that prevent the transition from the pre-fusion to post-fusion states. Here, we present cryo-EM analyses of a stabilized trimeric SARS-CoV S, as well as the trypsin-cleaved, stabilized S, and its interactions with ACE2. Neither binding to ACE2 nor cleavage by trypsin at the S1/S2 cleavage site impart large conformational changes within stabilized SARS-CoV S or expose the secondary cleavage site, S2′.

Template-primer binding affinity and RNase H cleavage specificity contribute to the strand transfer efficiency of HIV-1 reverse transcriptase

During reverse transcription of the HIV-1 genome, two strand-transfer events occur. Both events rely on the RNase H cleavage activity of reverse transcriptases (RTs) and template homology. Using a panel of mutants of HIV-1BH10 (group M/subtype B) and HIV-1ESP49 (group O) RTs and in vitro assays, we demonstrate that there is a strong correlation between RT minus-strand transfer efficiency and template-primer binding affinity. The highest strand transfer efficiencies were obtained with HIV-1ESP49 RT mutants containing the substitutions K358R/A359G/S360A, alone or in combination with V148I or T355A/Q357M. These HIV-1ESP49 RT mutants had been previously engineered to increase their DNA polymerase activity at high temperatures. Now, we found that RTs containing RNase H-inactivating mutations (D443N or E478Q) were devoid of strand transfer activity, whereas enzymes containing F61A or L92P had very low strand transfer activity. The strand transfer defect produced by L92P was attributed to a loss of template-primer binding affinity and, more specifically, to the higher dissociation rate constants (koff) shown by RTs bearing this substitution. Although L92P also deleteriously affected the RT's nontemplated nucleotide addition activity, neither nontemplated nucleotide addition activity nor the RT's clamp activities contributed to increased template switching when all tested mutant and WT RTs were considered. Interestingly, our results also revealed an association between efficient strand transfer and the generation of secondary cleavages in the donor RNA, consistent with the creation of invasion sites. Exposure of the elongated DNA at these sites facilitate acceptor (RNA or DNA) binding and promote template switching.

Mechanism and site requirements for thiophene hydrodesulfurization on supported Re domains in metal or sulfide form

The elementary steps and active structures involved in thiophene hydrodesulfurization (HDS) are examined here through structural and functional assessments of Re and ReSx catalysts prepared from ReOx precursors by treatment in H2 or H2S. These samples retain their respective bulk phases at sulfur chemical potentials prevalent during HDS, because nucleation barriers inhibit the interconversion of isotropic Re metal and lamellar ReSx layers. HDS turnover rates were much higher on ReSx than Re, but both phases showed similar kinetic effects of thiophene, H2, and H2S and binding constants for adsorbed thiophene and S-atoms, consistent with a common mechanism involving active sites that differ in number but not in binding properties. In such elementary steps, the surface consists of a template of refractory S-atoms that are bound irreversibly, known to form even at H2S/H2 ratios much lower than in HDS practice. Interstices within such templates can reversibly bind reactive intermediates, thus allowing catalytic turnovers, and act as HDS active sites. The number of such interstices depends on MS bond strength, which is lower for particles with ReSx than with Re bulk phases; their binding properties, however, are not dictated by the bulk phase, because they consist of those surface spaces that become capable of binding S-species weakly enough to allow their formation and removal as part of each catalytic turnover. On both Re and ReSx, thiophene conversion rates are limited by the addition of one H-atom to bound thiophene to form intermediate species that give tetrahydrothiophene (THT) and C4 hydrocarbons at a kinetic branch after this kinetically-relevant step. Thiophene pressures and H2S/H2 ratios do not influence THT/C4 product ratios, which decrease as residence time increases because of secondary CS cleavage in THT to form C4 products. Both products form in a single surface sojourn at similar site coverages by intermediates, as is also the case for secondary THT reactions. The effects of H2 on these primary and secondary events indicate that the kinetic branching occurs at a bound intermediate with the H-content of dihydrothiophene, from which the CS bond cleavage transition state is also formed. As in CC and CO cleavage, CS bond scission requires H-removal from saturated reactants (THT) by (i) increasing the bond order of its surface attachment; (ii) weakening the CX bond being cleaved (X = C, O, S); and (iii) evolving H2 to minimize entropy losses upon formation of the transition state.

Investigation on the microstructure and fracture behavior of super-eutectic ZA alloy welded joint by pulse fiber laser

In this study, we present the experimental laser welding of super-eutectic ZA alloy. Microstructures of the ZA alloy′s bond area welded by pulsed laser were investigated using an optical microscope, scanning electron microscope, energy diffraction spectrum and X-Ray diffractometer. The results showed that microstructures in the weld mainly consisted of threadlike columnar crystals, coarse dendrites and fine equiaxed grains. Secondary particles were produced in the interdendritic zones because of the composition segregation. A poor Al area was formed inside the η grain while a rich Al area takes shape outside the η grain. The occurrence of the composition segregation can increase the grain boundary cracking tendency. The existence of a large number of η phase greatly intensified the brittle of this material. The presence of many cleavage steps in the fractograph principally depended on the secondary cleavage effect between two cleavage cracks on different planes. The evolution of cracks along the twinning plane was the result of the secondary cleavage substituting for the twinning deformation in the vicinity of the crack tip.

Revisit of the anisotropic deformation behavior of single-crystal CaF2 in orthogonal cutting

A resolved stress model is presented to re-discuss the anisotropic deformation mechanism of single-crystal calcium fluoride in plunge-cut tests. The authors have investigated the machinability of CaF2 in ultra-precision cutting. However, the influence of crystal anisotropy on the brittle–ductile transition was discussed in a qualitative manner using optical microscopy and white light interferometer. Moreover, only the primary {100}<110> slip system and {111} cleavage were considered as deformation mechanisms. In this study, the ductility and brittleness of CaF2 is semi-quantitatively re-discussed on the basis of computation of the resolved stresses, by considering a secondary slip system and cleavage. The surface morphologies of the machined surface were characterized using field-emission scanning electron microscopy. The primary {100}<110> slip system is assumed to be dominant for the ductility. The brittle fracture initiated by the {111} cleavages was observed in a different manner in dependency rwith cutting directions.

Evaluation of high-cycle bending fatigue and fracture behaviors in EN-GJS700-2 ductile cast iron of crankshafts

Abstract In this article, high-cycle bending fatigue and fracture behaviors of the EN-GJS700-2 ductile cast iron, which has been widely utilized for crankshafts in engines, was experimentally evaluated. For such objective, standard specimens were extracted from the crankshaft and examined by the four-point rotary-bending high-cycle fatigue testing machine. Then, the S-N curve was evaluated for the studied material. Experiments were followed by the fracture investigation, using the scanning electron microscopy. Then, Obtained results indicated that high-cycle fatigue lifetimes were lower than the S-N curve from the FEMFAT data bank, since standard specimens were extracted from the crankshaft to consider the manufacturing effect. The failure analysis indicated different observations including striation marks (according to cyclic loading), debonding of nodular graphites from the ferritic-pearlitic matrix (as an evidence of the plastic deformation), micro-cracks and the secondary crack (near nodular graphites), cleavage marks (due to the brittle fracture), scratching marks (resulted from the friction and striking between two fracture surfaces) and inclusions.

Insights into the Structure of the Vip3Aa Insecticidal Protein by Protease Digestion Analysis.

Vip3 proteins are secretable proteins from Bacillus thuringiensis whose mode of action is still poorly understood. In this study, the activation process for Vip3 proteins was closely examined in order to better understand the Vip3Aa protein stability and to shed light on its structure. The Vip3Aa protoxin (of 89 kDa) was treated with trypsin at concentrations from 1:100 to 120:100 (trypsin:Vip3A, w:w). If the action of trypsin was not properly neutralized, the results of SDS-PAGE analysis (as well as those with Agrotis ipsilon midgut juice) equivocally indicated that the protoxin could be completely processed. However, when the proteolytic reaction was efficiently stopped, it was revealed that the protoxin was only cleaved at a primary cleavage site, regardless of the amount of trypsin used. The 66 kDa and the 19 kDa peptides generated by the proteases co-eluted after gel filtration chromatography, indicating that they remain together after cleavage. The 66 kDa fragment was found to be extremely resistant to proteases. The trypsin treatment of the protoxin in the presence of SDS revealed the presence of secondary cleavage sites at S-509, and presumably at T-466 and V-372, rendering C-terminal fragments of approximately 29, 32, and 42 kDa, respectively. The fact that the predicted secondary structure of the Vip3Aa protein shows a cluster of beta sheets in the C-terminal region of the protein might be the reason behind the higher stability to proteases compared to the rest of the protein, which is mainly composed of alpha helices.

Characterization of Lipid A Variants by Energy-Resolved Mass Spectrometry: Impact of Acyl Chains.

Lipid A molecules consist of a diglucosamine sugar core with a number of appended acyl chains that vary in their length and connectivity. Because of the challenging nature of characterizing these molecules and differentiating between isomeric species, an energy-resolved MS/MS strategy was undertaken to track the fragmentation trends and map genealogies of product ions originating from consecutive cleavages of acyl chains. Generalizations were developed based on the number and locations of the primary and secondary acyl chains as well as variations in preferential cleavages arising from the location of the phosphate groups. Secondary acyl chain cleavage occurs most readily for lipid A species at the 3' position, followed by primary acyl chain fragmentation at both the 3' and 3 positions. In the instances of bisphosphorylated lipid A variants, phosphate loss occurs readily in conjunction with the most favorable primary and secondary acyl chain cleavages. Graphical Abstract ᅟ.

Functional Characterization of Propeptides in Plant Subtilases as Intramolecular Chaperones and Inhibitors of the Mature Protease

Subtilisin-like serine proteases (SBTs) are extracellular proteases that depend on their propeptides for zymogen maturation and activation. The function of propeptides in plant SBTs is poorly understood and was analyzed here for the propeptide of tomato subtilase 3 (SBT3PP). SBT3PP was found to be required as an intramolecular chaperone for zymogen maturation and secretion of SBT3 in vivo Secretion was impaired in a propeptide-deletion mutant but could be restored by co-expression of the propeptide in trans SBT3 was inhibited by SBT3PP with a Kd of 74 nm for the enzyme-inhibitor complex. With a melting point of 87 °C, thermal stability of the complex was substantially increased as compared with the free protease suggesting that propeptide binding stabilizes the structure of SBT3. Even closely related propeptides from other plant SBTs could not substitute for SBT3PP as a folding assistant or autoinhibitor, revealing high specificity for the SBT3-SBT3PP interaction. Separation of the chaperone and inhibitor functions of SBT3PP in a domain-swap experiment indicated that they are mediated by different regions of the propeptide and, hence, different modes of interaction with SBT3. Release of active SBT3 from the autoinhibited complex relied on a pH-dependent cleavage of the propeptide at Asn-38 and Asp-54. The remarkable stability of the autoinhibited complex and pH dependence of the secondary cleavage provide means for stringent control of SBT3 activity, to ensure that the active enzyme is not released before it reaches the acidic environment of the trans-Golgi network or its final destination in the cell wall.

Revisiting the mechanism of coagulation factor XIII activation and regulation from a structure/functional perspective.

The activation and regulation of coagulation Factor XIII (FXIII) protein has been the subject of active research for the past three decades. Although discrete evidence exists on various aspects of FXIII activation and regulation a combinatorial structure/functional view in this regard is lacking. In this study, we present results of a structure/function study of the functional chain of events for FXIII. Our study shows how subtle chronological submolecular changes within calcium binding sites can bring about the detailed transformation of the zymogenic FXIII to its activated form especially in the context of FXIIIA and FXIIIB subunit interactions. We demonstrate what aspects of FXIII are important for the stabilization (first calcium binding site) of its zymogenic form and the possible modes of deactivation (thrombin mediated secondary cleavage) of the activated form. Our study for the first time provides a structural outlook of the FXIIIA2B2 heterotetramer assembly, its association and dissociation. The FXIIIB subunits regulatory role in the overall process has also been elaborated upon. In summary, this study provides detailed structural insight into the mechanisms of FXIII activation and regulation that can be used as a template for the development of future highly specific therapeutic inhibitors targeting FXIII in pathological conditions like thrombosis.

Distance effects during polyprotein processing in the complementation between defective FMDV RNAs

Passage of foot-and-mouth disease virus (FMDV) in BHK-21 cells resulted in the segmentation of the viral genome into two defective RNAs lacking part of either the L- or the capsid-coding region. The two RNAs are infectious by complementation. Electroporation of L-defective RNA in BHK-21 cells resulted in the accumulation of the precursor P3 located away from the deleted sequence. Expression of L in trans led to the processing of P3, indicating that there is a connection between L protease activity and the secondary cleavages carried out by 3C protease within P3. These results suggest that the complementation mechanism between defective RNAs is not restricted to supplying the L and capsid proteins but that distance effects on polyprotein processing events are also implicated.

Cleavage of Signal Regulatory Protein α (SIRPα) Enhances Inflammatory Signaling

Signal regulatory protein α (SIRPα) is a membrane glycoprotein immunoreceptor abundant in cells of monocyte lineage. SIRPα ligation by a broadly expressed transmembrane protein, CD47, results in phosphorylation of the cytoplasmic immunoreceptor tyrosine-based inhibitory motifs, resulting in the inhibition of NF-κB signaling in macrophages. Here we observed that proteolysis of SIRPα during inflammation is regulated by a disintegrin and metalloproteinase domain-containing protein 10 (ADAM10), resulting in the generation of a membrane-associated cleavage fragment in both THP-1 monocytes and human lung epithelia. We mapped a charge-dependent putative cleavage site near the membrane-proximal domain necessary for ADAM10-mediated cleavage. In addition, a secondary proteolytic cleavage within the membrane-associated SIRPα fragment by γ-secretase was identified. Ectopic expression of a SIRPα mutant plasmid encoding a proteolytically resistant form in HeLa cells inhibited activation of the NF-κB pathway and suppressed STAT1 phosphorylation in response to TNFα to a greater extent than expression of wild-type SIRPα. Conversely, overexpression of plasmids encoding the proteolytically cleaved SIRPα fragments in cells resulted in enhanced STAT-1 and NF-κB pathway activation. Thus, the data suggest that combinatorial actions of ADAM10 and γ-secretase on SIRPα cleavage promote inflammatory signaling.

Temperature micro-mapping in oscillatory-zoned chlorite: Application to study of a green-schist facies fault zone in the Pyrenean Axial Zone (Spain)

Oscillatory compositional zoning in minerals has been observed in hydrothermal, magmatic, and metamorphic environments and is commonly attributed to chemical or physical cyclical changes during crystal growth. Chemical zoning is a common feature of solid solutions, which has been rarely reported in phyllosilicates. In this study, oscillatory zoning in chlorite is described in samples from the Pic-de-Port-Vieux thrust, a minor thrust fault associated to the major Gavarnie thrust fault zone (Central Pyrenees, Spain). The Pic-de-Port-Vieux thrust sheet comprises a 1–20 m thick layer of Triassic red pelite and sandstone thrust over mylonitized Cretaceous dolomitic limestone. The thrust fault zone deformation comprises secondary faults and cleavage affecting the Triassic pelite and sandstone. An important feature responsible to this deformation is a set of veins filled by quartz and chlorite. Chlorite is present in crack-seal extension veins and in shear veins; both structures opened under the same stress conditions. In some shear veins, chlorite occurs as pseudo-uniaxial plates arranged in rosette-shaped aggregates. These aggregates appear to have developed as a result of radial growth of the chlorite platelets. Oscillatory zoning has been imaged by backscattered scanning electron microscopy and by X-ray quantitative micro-mapping. These oscillations correspond to chemical zoning with alternating iron-rich and magnesium-rich bands. The chlorite composition ranges from a Fe-rich pole to a Mg-rich pole. Fe 3+ /∑Fe values were measured in chlorite using μ-XANES spot analyses and vary from 0.23 to 0.44. The highest values are in the Fe-rich area. Temperature maps, built from standardized microprobe X-ray images and redox state using the program XMapTools, indicate oscillatory variations from about 310 to 400 ± 50 °C during chlorite crystallization. These temperature variations are correlated with a Fe 3+ /∑Fe variation by Al 3+ Fe 3+ −1 and di-trioctahedral substitutions highlighted by Mg and Fe Tot contents (Fe-Mg zoning). Chemical variations could be then explained by alternation of cooling times and cyclical pulses of a fluid hotter than the host rock. It is however not excluded that kinetic effects influence the incorporation of Mg or Fe during chlorite crystallization.