Thermo-Mechanical Fatigue Crack Growth and Phase Angle Effects in Ti6246.

A bespoke TMF crack growth test set-up has been developed and validated for use throughout this study and the effects of phasing between mechanical loading and temperature have been investigated. The study shows that TMF cycles may show increased crack growth rate behaviour when compared to isothermal fatigue. The phase angle of the applied TMF cycle can also affect crack growth behaviour, with in-phase (IP) test conditions showing faster crack growth rates than out-of-phase (OP) test conditions. Propagating cracks interact with the microstructure of the material, in particular, the α/ß interfaces within the prior beta grains and supporting fractography evidences subtle differences in fracture mechanisms as a result of phase angle.

Targeting Tumor Angiogenesis with the Selective VEGFR-3 Inhibitor EVT801 in Combination with Cancer Immunotherapy.

The receptor tyrosine kinase VEGFR-3 plays a crucial role in cancer-induced angiogenesis and lymphangiogenesis, promoting tumor development and metastasis. Here, we report the novel VEGFR-3 inhibitor EVT801 that presents a more selective and less toxic profile than two major inhibitors of VEGFRs (i.e., sorafenib and pazopanib). As monotherapy, EVT801 showed a potent antitumor effect in VEGFR-3-positive tumors, and in tumors with VEGFR-3-positive microenvironments. EVT801 suppressed VEGF-C-induced human endothelial cell proliferation in vitro and tumor (lymph)angiogenesis in different tumor mouse models. In addition to reduced tumor growth, EVT801 decreased tumor hypoxia, favored sustained tumor blood vessel homogenization (i.e., leaving fewer and overall larger vessels), and reduced important immunosuppressive cytokines (CCL4, CCL5) and myeloid-derived suppressor cells (MDSC) in circulation. Furthermore, in carcinoma mouse models, the combination of EVT801 with immune checkpoint therapy (ICT) yielded superior outcomes to either single treatment. Moreover, tumor growth inhibition was inversely correlated with levels of CCL4, CCL5, and MDSCs after treatment with EVT801, either alone or combined with ICT. Taken together, EVT801 represents a promising anti(lymph)angiogenic drug for improving ICT response rates in patients with VEGFR-3 positive tumors. Significance: The VEGFR-3 inhibitor EVT801 demonstrates superior selectivity and toxicity profile than other VEGFR-3 tyrosine kinase inhibitors. EVT801 showed potent antitumor effects in VEGFR-3-positive tumors, and tumors with VEGFR-3-positive microenvironments through blood vessel homogenization, and reduction of tumor hypoxia and limited immunosuppression. EVT801 increases immune checkpoint inhibitors' antitumor effects.

Detecting substrate glycans of fucosyltransferases with fluorophore-conjugated fucose and methods for glycan electrophoresis.

Like sialylation, fucose usually locates at the nonreducing ends of various glycans on glycoproteins and constitutes important glycan epitopes. Detecting the substrate glycans of fucosyltransferases is important for understanding how these glycan epitopes are regulated in response to different growth conditions and external stimuli. Here we report the detection of these glycans on glycoproteins as well as in their free forms via enzymatic incorporation of fluorophore-conjugated fucose using FUT2, FUT6, FUT7, FUT8 and FUT9. Specifically, we describe the detection of the substrate glycans of these enzymes on fetal bovine fetuin, recombinant H1N1 viral neuraminidase and therapeutic antibodies. The detected glycans include complex and high-mannose N-glycans. By establishing a series of precursors for the synthesis of Lewis X and sialyl Lewis X structures, we not only provide convenient electrophoresis methods for studying glycosylation but also demonstrate the substrate specificities and some kinetic features of these enzymes. Our results support the notion that fucosyltransferases are key targets for regulating the synthesis of Lewis X and sialyl Lewis X structures.

The Effect of Phase Angle on the Thermo-Mechanical Fatigue Life of a Titanium Metal Matrix Composite.

The thermo-mechanical fatigue (TMF) behaviour of a Ti-6Al-4V matrix composite reinforced with SCS-6 silicon carbide fibres (140 µm longitudinal fibres, laid up hexagonally) has been investigated. In-phase and out-of-phase TMF cycles were utilized, cycling between 80⁻300 °C, with varying maximum stress. The microstructure and fracture surfaces were studied using electron backscatter diffraction (EBSD), energy-dispersive X-ray spectroscopy (EDS), scanning electron microscopy (SEM), profilometry, and optical microscopy. The results have shown the damaging effect of out-of-phase cycling with crack initiation occurring earlier than in in-phase tests and crack propagation rates being accelerated in out-of-phase cycles. Fatigue crack initiation has been shown to be sensitive to crystallographic texture in the cladding material and thermo-mechanical fatigue test results can be considered according to a previously proposed conceptual framework for the interpretation of metal matrix composite fatigue.

Creep Deformation by Dislocation Movement in Waspaloy.

Creep tests of the polycrystalline nickel alloy Waspaloy have been conducted at Swansea University, for varying stress conditions at 700 °C. Investigation through use of Transmission Electron Microscopy at Cambridge University has examined the dislocation networks formed under these conditions, with particular attention paid to comparing tests performed above and below the yield stress. This paper highlights how the dislocation structures vary throughout creep and proposes a dislocation mechanism theory for creep in Waspaloy. Activation energies are calculated through approaches developed in the use of the recently formulated Wilshire Equations, and are found to differ above and below the yield stress. Low activation energies are found to be related to dislocation interaction with γ' precipitates below the yield stress. However, significantly increased dislocation densities at stresses above yield cause an increase in the activation energy values as forest hardening becomes the primary mechanism controlling dislocation movement. It is proposed that the activation energy change is related to the stress increment provided by work hardening, as can be observed from Ti, Ni and steel results.

Development and Assessment of a New Empirical Model for Predicting Full Creep Curves.

This paper details the development and assessment of a new empirical creep model that belongs to the limited ranks of models reproducing full creep curves. The important features of the model are that it is fully standardised and is universally applicable. By standardising, the user no longer chooses functions but rather fits one set of constants only. Testing it on 7 contrasting materials, reproducing 181 creep curves we demonstrate its universality. New model and Theta Projection curves are compared to one another using an assessment tool developed within this paper.

Altered colonic mucosal availability of n-3 and n-6 polyunsaturated fatty acids in ulcerative colitis and the relationship to disease activity.

BACKGROUND AND AIMS: The polyunsaturated fatty acids (PUFA) arachidonic acid (AA, n-6) and eicosapentaenoic acid (EPA, n-3) are precursors of eicosanoids and other lipid mediators which have critical roles in inflammation. The mediators formed from the different PUFA have different potencies. We hypothesised that metabolic changes associated with colonic mucosal inflammation would modify the bioavailability of the eicosanoid precursors AA and EPA. METHODS: Colonic mucosa biopsies were obtained from patients with ulcerative colitis and from matched controls. Inflammation was graded endoscopically and histologically. Esterified and non-esterified fatty acids were determined within the biopsies using gas chromatography-mass spectrometry and liquid chromatography-mass spectrometry, respectively. RESULTS: Biopsy samples were collected from 69 UC patients (54 providing both inflamed and non-inflamed mucosa) and 69 controls. Inflamed mucosa had higher AA (p<0.001) and lower EPA (p<0.010) contents and a higher AA:EPA ratio (p<0.001). Inflamed mucosa also had higher docosapentaenoic acid (DPA) and docosahexaenoic acid (DHA) and lower linoleic acid (LA) and α-linolenic acid (α-LNA) contents (all p<0.001), compared to non-inflamed and controls. There were significant correlations between severity of inflammation and contents of AA, DPA and DHA (positive correlations) and of LA, α-LNA and EPA (negative correlations). CONCLUSIONS: Higher AA, AA:EPA ratio, DPA and DHA and lower LA, α-LNA and EPA are seen in inflamed mucosa in UC and correlate with severity of inflammation. This suggests an alteration in fatty acid metabolism in the inflamed gut mucosa, which may offer novel targets for intervention and should be considered if nutritional strategies are used.

A Model for Creep and Creep Damage in the γ-Titanium Aluminide Ti-45Al-2Mn-2Nb.

Gamma titanium aluminides (γ-TiAl) display significantly improved high temperature mechanical properties over conventional titanium alloys. Due to their low densities, these alloys are increasingly becoming strong candidates to replace nickel-base superalloys in future gas turbine aeroengine components. To determine the safe operating life of such components, a good understanding of their creep properties is essential. Of particular importance to gas turbine component design is the ability to accurately predict the rate of accumulation of creep strain to ensure that excessive deformation does not occur during the component's service life and to quantify the effects of creep on fatigue life. The theta (θ) projection technique is an illustrative example of a creep curve method which has, in this paper, been utilised to accurately represent the creep behaviour of the γ-TiAl alloy Ti -45Al-2Mn-2Nb. Furthermore, a continuum damage approach based on the θ-projection method has also been used to represent tertiary creep damage and accurately predict creep rupture.

A Critical Analysis of the Conventionally Employed Creep Lifing Methods.

The deformation of structural alloys presents problems for power plants and aerospace applications due to the demand for elevated temperatures for higher efficiencies and reductions in greenhouse gas emissions. The materials used in such applications experience harsh environments which may lead to deformation and failure of critical components. To avoid such catastrophic failures and also increase efficiency, future designs must utilise novel/improved alloy systems with enhanced temperature capability. In recognising this issue, a detailed understanding of creep is essential for the success of these designs by ensuring components do not experience excessive deformation which may ultimately lead to failure. To achieve this, a variety of parametric methods have been developed to quantify creep and creep fracture in high temperature applications. This study reviews a number of well-known traditionally employed creep lifing methods with some more recent approaches also included. The first section of this paper focuses on predicting the long-term creep rupture properties which is an area of interest for the power generation sector. The second section looks at pre-defined strains and the re-production of full creep curves based on available data which is pertinent to the aerospace industry where components are replaced before failure.

Fighting obesity with a sugar-based library: discovery of novel MCH-1R antagonists by a new computational-VAST approach for exploration of GPCR binding sites.

Obesity is an increasingly common disease. While antagonism of the melanin-concentrating hormone-1 receptor (MCH-1R) has been widely reported as a promising therapeutic avenue for obesity treatment, no MCH-1R antagonists have reached the market. Discovery and optimization of new chemical matter targeting MCH-1R is hindered by reduced HTS success rates and a lack of structural information about the MCH-1R binding site. X-ray crystallography and NMR, the major experimental sources of structural information, are very slow processes for membrane proteins and are not currently feasible for every GPCR or GPCR-ligand complex. This situation significantly limits the ability of these methods to impact the drug discovery process for GPCR targets in "real-time", and hence, there is an urgent need for other practical and cost-efficient alternatives. We present here a conceptually pioneering approach that integrates GPCR modeling with design, synthesis, and screening of a diverse library of sugar-based compounds from the VAST technology (versatile assembly on stable templates) to provide structural insights on the MCH-1R binding site. This approach creates a cost-efficient new avenue for structure-based drug discovery (SBDD) against GPCR targets. In our work, a primary VAST hit was used to construct a high-quality MCH-1R model. Following model validation, a structure-based virtual screen yielded a 14% hit rate and 10 novel chemotypes of potent MCH-1R antagonists, including EOAI3367472 (IC50 = 131 nM) and EOAI3367474 (IC50 = 213 nM).

Cytokine mucosal expression in ulcerative colitis, the relationship between cytokine release and disease activity.

BACKGROUND: Ulcerative colitis (UC) is an inflammatory bowel disease with conflicting evidence from studies on the roles of TNFα, IL-8, TGFß and other cytokines and characterised by neutrophil infiltration and tissue destruction. AIM: To compare cytokine profiles of inflamed and non-inflamed mucosa in patients with distal UC, and matched controls. METHODS: Patients were prospectively recruited, mucosal biopsies at flexible sigmoidoscopy (FS) were taken from UC patients within macroscopically inflamed and non-inflamed proximal mucosa, and from age-sex matched controls undergoing FS. Endoscopic and histological inflammation was graded. Quantitative cytokine analysis for IL-4, TNFα, IL-17A, IL-8, IL-10, TGFß and IFNγ was carried out on tissue homogenates. Statistical comparison was by Wilcoxon signed rank pair analysis, Mann-Whitney U test and Spearman's correlation. RESULTS: 69 active UC patients (54 paired non-inflamed/inflamed mucosa) and 69 controls were compared. In inflamed mucosa, elevation in IL-8 and reduction in TGFß was measured compared with non-inflamed mucosa (p<0.001; p<0.02) and control mucosa (p<0.001; p<0.001); IL-8 was positively correlated (rs=0.481, p<0.01) and TGFß inversely correlated (rs=0.462; p<0.01) with grade of inflammation. TNFα concentration was not significantly different. Comparisons of inflamed with non-inflamed mucosa also demonstrate significant reduction in concentration of IFNγ (p<0.001), IL-4 (p<0.005) and IL-17A (p<0.002). CONCLUSION: Our findings suggest that IL-8 is elevated and TGFß is reduced in distal colitis. Lower concentration of IFNγ, IL-4 and IL-17A were also noted. TNFα levels were unchanged. These findings suggest that the inflammatory response in UC may predominantly involve IL-8 mediated neutrophil infiltration and failure of TGFß mediated tissue healing.

An Empirical Approach to Correlating Thermo-Mechanical Fatigue Behaviour of a Polycrystalline Ni-Base Superalloy.

Assessment of thermo-mechanical fatigue behaviour of the polycrystalline nickel alloy RR1000 reveals a significant effect of phase angle on fatigue life. The current paper explores two scenarios: the first where the mechanical strain range is held constant and comparisons of the fatigue life are made for different phase angle tests; and secondly, the difference between the behaviour of In-phase (IP) and -180° Out-Of-Phase (OOP) tests over a variety of applied strain ranges. It is shown that different lifing approaches are currently required for the two scenarios, with a mean stress based approach being more applicable in the first case, whereas a Basquin-type model proves more applicable in the second. However, it is also demonstrated that the crack propagation phase should also be considered in these types of tests for high strain ranges and projects that future modelling approaches should attempt to unify mean stress, stress range and a crack propagation phase.

Recent Advances in Creep Modelling of the Nickel Base Superalloy, Alloy 720Li.

Recent work in the creep field has indicated that the traditional methodologies involving power law equations are not sufficient to describe wide ranging creep behaviour. More recent approaches such as the Wilshire equations however, have shown promise in a wide range of materials, particularly in extrapolation of short term results to long term predictions. In the aerospace industry however, long term creep behaviour is not critical and more focus is required on the prediction of times to specific creep strains. The current paper illustrates the capability of the Wilshire equations to recreate full creep curves in a modern nickel superalloy. Furthermore, a finite-element model based on this method has been shown to accurately predict stress relaxation behaviour allowing more accurate component lifing.

TBAF-catalysed silver oxide-mediated cross-coupling of functional trimethysilylpyridines: access to arylpyridines and bihetaryl compounds.

The concomitant use of silver oxide and catalytic amount of TBAF allowed the efficient and chemoselective coupling of readily available 4-chloro- and 4-methyl-2-trimethyl-silyl-pyridines with heteroaromatic and aromatic halides. Based on control experiments, a mechanism involving the formation of a pyridylsilver intermediate and TBAF recycling is postulated.

Prediction of cyclin-dependent kinase 2 inhibitor potency using the fragment molecular orbital method.

BACKGROUND: The reliable and robust estimation of ligand binding affinity continues to be a challenge in drug design. Many current methods rely on molecular mechanics (MM) calculations which do not fully explain complex molecular interactions. Full quantum mechanical (QM) computation of the electronic state of protein-ligand complexes has recently become possible by the latest advances in the development of linear-scaling QM methods such as the ab initio fragment molecular orbital (FMO) method. This approximate molecular orbital method is sufficiently fast that it can be incorporated into the development cycle during structure-based drug design for the reliable estimation of ligand binding affinity. Additionally, the FMO method can be combined with approximations for entropy and solvation to make it applicable for binding affinity prediction for a broad range of target and chemotypes. RESULTS: We applied this method to examine the binding affinity for a series of published cyclin-dependent kinase 2 (CDK2) inhibitors. We calculated the binding affinity for 28 CDK2 inhibitors using the ab initio FMO method based on a number of X-ray crystal structures. The sum of the pair interaction energies (PIE) was calculated and used to explain the gas-phase enthalpic contribution to binding. The correlation of the ligand potencies to the protein-ligand interaction energies gained from FMO was examined and was seen to give a good correlation which outperformed three MM force field based scoring functions used to appoximate the free energy of binding. Although the FMO calculation allows for the enthalpic component of binding interactions to be understood at the quantum level, as it is an in vacuo single point calculation, the entropic component and solvation terms are neglected. For this reason a more accurate and predictive estimate for binding free energy was desired. Therefore, additional terms used to describe the protein-ligand interactions were then calculated to improve the correlation of the FMO derived values to experimental free energies of binding. These terms were used to account for the polar and non-polar solvation of the molecule estimated by the Poisson-Boltzmann equation and the solvent accessible surface area (SASA), respectively, as well as a correction term for ligand entropy. A quantitative structure-activity relationship (QSAR) model obtained by Partial Least Squares projection to latent structures (PLS) analysis of the ligand potencies and the calculated terms showed a strong correlation (r2 = 0.939, q2 = 0.896) for the 14 molecule test set which had a Pearson rank order correlation of 0.97. A training set of a further 14 molecules was well predicted (r2 = 0.842), and could be used to obtain meaningful estimations of the binding free energy. CONCLUSIONS: Our results show that binding energies calculated with the FMO method correlate well with published data. Analysis of the terms used to derive the FMO energies adds greater understanding to the binding interactions than can be gained by MM methods. Combining this information with additional terms and creating a scaled model to describe the data results in more accurate predictions of ligand potencies than the absolute values obtained by FMO alone.

Compound Design by Fragment-Linking.

The linking together of two fragment compounds that bind to distinct protein sub-sites can lead to a superadditivity of binding affinities, in which the binding free energy of the linked fragments exceeds the simple sum of the binding energies of individual fragments (linking coefficient E<1). However, a review of the literature shows that such events are relatively rare and, in the majority of the cases, linking coefficients are far from optimal being much greater than 1. It is critical to design a linker that does not disturb the original binding poses of each fragment in order to achieve successful linking. However, such an ideal linker is often difficult to design and even more difficult to actually synthesize. We suggest that the chance of achieving successful fragment linking can be significantly improved by choosing a fragment pair that consists of one fragment that binds by strong H-bonds (or non-classical equivalents) and a second fragment that is more tolerant of changes in binding mode (hydrophobic or vdW binders). We also propose that the fragment molecular orbital (FMO) calculations can be used to analyse the nature of the binding interactions of the fragment hits for the selection of fragments for evolution, merging and linking in order to optimize the chance of success.

Novel 6,7,8,9-tetrahydro-5H-1,4,7,10a-tetraaza-cyclohepta[f]indene analogues as potent and selective 5-HT(2C) agonists for the treatment of metabolic disorders.

The discovery of a novel series of 5-HT(2C) agonists based on a tricyclic pyrazolopyrimidine scaffold is described. Compounds with good levels of in vitro potency and moderate to good levels of selectivity with respect to the 5-HT(2A) and 5-HT(2B) receptors were identified. One of the analogues (7 g) was found to be efficacious in a sub-chronic weight loss model. A key limitation of the series of compounds was that they were found to be potent inhibitors of the hERG ion channel. Some compounds, bearing polar side chains were identified which showed a much reduced hERG liability however these compounds were sub-optimal in terms of their in vitro potency or selectivity.

An orally bioavailable positive allosteric modulator of the mGlu4 receptor with efficacy in an animal model of motor dysfunction.

A high-throughput screening campaign identified 4-((E)-styryl)-pyrimidin-2-ylamine (11) as a positive allosteric modulator of the metabotropic glutamate (mGlu) receptor subtype 4. An evaluation of the structure-activity relationships (SAR) of 11 is described and the efficacy of this compound in a haloperidol-induced catalepsy rat model following oral administration is presented.