Assessment of EGFR/HER2 dimerization by FRET-FLIM utilizing Alexa-conjugated secondary antibodies in relation to targeted therapies in cancers

The expression level of the HER family is unreliable as a predictive marker for targeted therapies in cancer. Thus, there is a need to develop other biomarkers, which can be used to accurately select responsive patients for targeted therapies. The HER dimerization status may be more important than HER receptor expression per se in determining sensitivity or resistance to a given therapeutic agent. The aim of the study is to develop a FRET assay using dye conjugated secondary antibodies to assess HER receptor dimerization. Using primary antibodies from different species in conjunction with Alexa488 and Alexa546 conjugated secondary antibodies, we validated our EGFR/HER2 dimerization assay in three cell lines, EGFR positive A431 cells as well as HER2 positive breast cell lines BT474 and SKBR3 cells. Finally, we applied our assay to assess EGFR/HER2 dimerization in paraffin embedded cell pellets. Our results show promise for the assay to be applied to tumor samples in order to assess the prognostic significance and predictive value of HER receptor dimerization in various cancers

Energy and force analysis of Ti-6Al-4V linear friction welds for computational modeling input and validation data

The linear friction welding (LFW) process is finding increasing use as a manufacturing technology for the production of titanium alloy Ti-6Al-4V aerospace components. Computational models give an insight into the process, however, there is limited experimental data that can be used for either modeling inputs or validation. To address this problem, a design of experiments approach was used to investigate the influence of the LFW process inputs on various outputs for experimental Ti-6Al-4V welds. The finite element analysis software DEFORM was also used in conjunction with the experimental findings to investigate the heating of the workpieces. Key findings showed that the average interface force and coefficient of friction during each phase of the process were insensitive to the rubbing velocity; the coefficient of friction was not coulombic and varied between 0.3 and 1.3 depending on the process conditions; and the interface of the workpieces reached a temperature of approximately approximately 1273 K (1000 °C) at the end of phase 1. This work has enabled a greater insight into the underlying process physics and will aid future modeling investigations.EPSRC, Boeing Company, Welding Institut

Regularization-independent study of renormalized non-perturbative quenched QED

A recently proposed regularization-independent method is used for the first time to solve the renormalized fermion Schwinger-Dyson equation numerically in quenched QED$_4$. The Curtis-Pennington vertex is used to illustrate the technique and to facilitate comparison with previous calculations which used the alternative regularization schemes of modified ultraviolet cut-off and dimensional regularization. Our new results are in excellent numerical agreement with these, and so we can now conclude with confidence that there is no residual regularization dependence in these results. Moreover, from a computational point of view the regularization independent method has enormous advantages, since all integrals are absolutely convergent by construction, and so do not mix small and arbitrarily large momentum scales. We analytically predict power law behaviour in the asymptotic region, which is confirmed numerically with high precision. The successful demonstration of this efficient new technique opens the way for studies of unquenched QED to be undertaken in the near future.Comment: 20 pages,5 figure

Renormalization and Chiral Symmetry Breaking in Quenched QED in Arbitrary Covariant Gauge

We extend a previous Landau-gauge study of subtractive renormalization of the fermion propagator Dyson-Schwinger equation (DSE) in strong-coupling, quenched QED_4 to arbitrary covariant gauges. We use the fermion-photon proper vertex proposed by Curtis and Pennington with an additional correction term included to compensate for the small gauge-dependence induced by the ultraviolet regulator. We discuss the chiral limit and the onset of dynamical chiral symmetry breaking in the presence of nonperturbative renormalization. We extract the critical coupling in several different gauges and find evidence of a small residual gauge-dependence in this quantity.Comment: REVTEX 3.0, 27 pages including 14 Extended Postscript files comprising 9 figures. Replacement: discussion of chiral limit corrected, and some minor typographical errors fixed. To appear in Phys. Rev.

HER2-HER3 heterodimer quantification by FRET-FILM and patient subclass analysis of the COIN colorectal trial

BACKGROUND: The phase 3 MRC COIN trial showed no statistically significant benefit from adding the EGFR-target cetuximab to oxaliplatin-based chemotherapy in first-line treatment of advanced colorectal cancer. This study exploits additional information on HER2-HER3 dimerization to achieve patient stratification and reveal previously hidden subgroups of patients who had differing disease progression and treatment response. METHODS: HER2-HER3 dimerization was quantified by 'FLIM Histology' in primary tumor samples from 550 COIN trial patients receiving oxaliplatin and fluoropyrimidine chemotherapy +/-cetuximab. Bayesian latent class analysis (LCA) and covariate reduction was performed to analyze the effects of HER2-HER3 dimer, RAS mutation and cetuximab on progression-free survival (PFS) and overall survival (OS). All statistical tests were two-sided. RESULTS: LCA on a cohort of 398 patients revealed two patient subclasses with differing prognoses (median OS: 1624 days [95%CI=1466-1816] vs 461 [95%CI=431-504]): Class 1 (15.6%) showed a benefit from cetuximab in OS (HR = 0.43 [95%CI=0.25-0.76]; p = 0.004). Class 2 showed an association of increased HER2-HER3 with better OS (HR = 0.64 [95%CI=0.44-0.94]; p = 0.02). A class prediction signature was formed and tested on an independent validation cohort (N = 152) validating the prognostic utility of the dimer assay. Similar subclasses were also discovered in full trial dataset (N = 1,630) based on 10 baseline clinicopathological and genetic covariates. CONCLUSIONS: Our work suggests that the combined use of HER dimer imaging and conventional mutation analyses will be able to identify a small subclass of patients (>10%) who will have better prognosis following chemotherapy. A larger prospective cohort will be required to confirm its utility in predicting the outcome of anti-EGFR treatment

Putting context to numbers : a geotechnical risk trajectory to cost overrun extremism

The study investigates the cause of the unusually high cost overruns experienced in highway project delivery in the tropical wetland setting of the Niger Delta region of Nigeria. This is in view of the extensive literature supporting the link between geology, the lack of geotechnical best practices and cost overruns. An empirical profiling of cost overrun research further reveals the predominance of mono-method studies based on survey methods, correlative analysis and archival data modelling techniques, all of which are underlain by positivism. The study argues that such positivist philosophies, although methodologically valid, cannot adequately explain and provide in-depth understanding of the contextual cost overrun drivers in highway organisations., Using a robust and thoughtfully designed mix of methods, the paper examines the contribution of geotechnical risks to cost overruns experienced in highway project, and demonstrates the relevance of context in cost overrun research. Cost overrun data from documentary sources for 61 completed highway projects in the Niger Delta are gathered and analysed, revealing an average value of 216%, with extreme cases, ranging up to 1925% of budgeted cost. To uncover the intrinsic contextual drivers, 16 interviews were conducted with participants from the three highway agencies in the region, responsible for the execution of the sampled highway projects. Adopting a geotechnical narrative, the data is thematically analysed, deductively and inductively. The results of the analysis identified that poor project governance, management and procurement practices, have inhibited the competent management of geotechnical risk, creating a propensity for extreme cost overruns on the highway projects. The study submits the phenomenon of cost overruns in public infrastructure projects is underlain by a complexity of contextual social constructs, which would have been overlooked in positivists studies. Cost overrun research therefore, needs to be contextually and numerically anchored. Keywords: Context, Cost overruns, Highway projects, Mixed methods, Social Construct

On the verge of Umdeutung in Minnesota: Van Vleck and the correspondence principle (Part One)

In October 1924, the Physical Review, a relatively minor journal at the time, published a remarkable two-part paper by John H. Van Vleck, working in virtual isolation at the University of Minnesota. Van Vleck combined advanced techniques of classical mechanics with Bohr's correspondence principle and Einstein's quantum theory of radiation to find quantum analogues of classical expressions for the emission, absorption, and dispersion of radiation. For modern readers Van Vleck's paper is much easier to follow than the famous paper by Kramers and Heisenberg on dispersion theory, which covers similar terrain and is widely credited to have led directly to Heisenberg's "Umdeutung" paper. This makes Van Vleck's paper extremely valuable for the reconstruction of the genesis of matrix mechanics. It also makes it tempting to ask why Van Vleck did not take the next step and develop matrix mechanics himself.Comment: 82 page

Measurement of the B0-anti-B0-Oscillation Frequency with Inclusive Dilepton Events

The $B^0$-$\bar B^0$ oscillation frequency has been measured with a sample of 23 million \B\bar B pairs collected with the BABAR detector at the PEP-II asymmetric B Factory at SLAC. In this sample, we select events in which both B mesons decay semileptonically and use the charge of the leptons to identify the flavor of each B meson. A simultaneous fit to the decay time difference distributions for opposite- and same-sign dilepton events gives $\Delta m_d = 0.493 \pm 0.012{(stat)}\pm 0.009{(syst)}$ ps$^{-1}$.Comment: 7 pages, 1 figure, submitted to Physical Review Letter

Genetic predisposition to ductal carcinoma in situ of the breast

Background: Ductal carcinoma in situ (DCIS) is a non-invasive form of breast cancer. It is often associated with invasive ductal carcinoma (IDC), and is considered to be a non-obligate precursor of IDC. It is not clear to what extent these two forms of cancer share low-risk susceptibility loci, or whether there are differences in the strength of association for shared loci. Methods: To identify genetic polymorphisms that predispose to DCIS, we pooled data from 38 studies comprising 5,067 cases of DCIS, 24,584 cases of IDC and 37,467 controls, all genotyped using the iCOGS chip. Results: Most (67 %) of the 76 known breast cancer predisposition loci showed an association with DCIS in the same direction as previously reported for invasive breast cancer. Case-only analysis showed no evidence for differences between associations for IDC and DCIS after considering multiple testing. Analysis by estrogen receptor (ER) status confirmed that loci associated with ER positive IDC were also associated with ER positive DCIS. Analysis of DCIS by grade suggested that two independent SNPs at 11q13.3 near CCND1 were specific to low/intermediate grade DCIS (rs75915166, rs554219). These associations with grade remained after adjusting for ER status and were also found in IDC. We found no novel DCIS-specific loci at a genome wide significance level of P < 5.0x10-8. Conclusion: In conclusion, this study provides the strongest evidence to date of a shared genetic susceptibility for IDC and DCIS. Studies with larger numbers of DCIS are needed to determine if IDC or DCIS specific loci exist