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PDF file 70K, This table contains the IC50 values of in vitro cell survival assay for crizotinib or ceritinib in engineered EML4-ALK mutant Ba/F3 cells corresponding to Fig 4A

PDF file 135K, Figure S1 contains data about Ceritinib specificity and potency on crizotinib na?ve EML4-ALK wild-type cells. Figure S2 contains the Cell survival curves of H3122 CR1, MGH021-4 and MGH045 cells following treatment with Crizotinib or ceritinib corresponding to Fig2A. Figure S3 contains the Cell survival curves of Ba/F3 cells following treatment with Crizotinib or ceritinib corresponding to Fig4A. Figure S4 contains tumor growth curves corresponding to the Development of crizotinib resistant H2228 xenograft tumor models used in Fig 5A-D

PDF file 100K, Legends for the supplementary figures

An after etch overlay measurement on device is typically used as a reference overlay as this is what determines the final overlay. The delta between on target overlay from after develop (ADI) and this reference overlay on device after etch (AEI) is known as the metrology to device offset (MTD). As the fab overlay is controlled by a run-to-run control of ADI overlay, it is preferred to minimize the MTD. The MTD concept in overlay metrology has long been present in the industry and many ways to mitigate this problem have been adopted (such as designing overlay target at ADI that has a similarly low aberration response as the device, or dialing in a static offset between ADI and AEI overlay, etc.). As overlay margins continue to shrink, existing methods start to show gaps and are insufficient to suppress the MTD to an acceptable level on the few most critical overlay layers. In order to address this gap, we need to deploy a much wider solution space that provides an integrated design-lithography-etch solution. In order to characterize the MTD, (assuming that target design in ADI has already minimized aberration response delta between target and device), it is important to understand that there are two major components to MTD: (1) an inaccuracy in ADI overlay (metrology artifacts mostly due to the presence of target asymmetry) and (2) etch to litho offset due to any post ADI added effects such as etch induced expansion and/or stress release etc. However, the above two components are strongly coupled and traditional characterization methods have difficulty in separating their contribution to the measurement. In this technical paper we will discuss novel methods (data driven as well as model-based) to decouple these and multi-lot results will show that MTD can be further minimized compared to traditional static correction between ADI and AEI.

An urban stormwater retention pond was instrumented with temperature (T) and dissolved oxygen (DO) sensors connected to wireless sensor networks (WSNs) for providing real-time water quality...

N-substituted azaindoles were discovered as potent pan-PIM inhibitors. Lead optimization, guided by structure and focused on physico-chemical properties allowed us to solve inherent hERG and permeability liabilities, and provided compound 27, which subsequently impacted KG-1 tumor growth in a mouse model.

N-substituted azaindoles were discovered as promising pan-PIM inhibitors. Lead optimization is described en route toward the identification of a clinical candidate. Modulation of physico-chemical properties allowed to solve inherent hERG and permeability liabilities. Compound 17 showed tumor growth inhibition in a KG1 tumor-bearing mouse model.

The Visible Infrared Imager Radiometer Suite (VIIRS) Reflective Solar Band Automated Calibration (RSBAutoCal) Day Night Band (DNB) dark signal and gain ratio calculations exhibit two issues: gain ratio trending instability and gain ratio differences relative to those calculated using the VIIRS Recommended Operating Procedures (VROP) monthly look-up table (LUT) generation tool. These issues have thus far prevented transition of RSBAutoCal to automated mode for these two DNB calibration parameters. In this paper we describe the successful diagnosis of the gain ratio stability issue. We also present a solution to stabilize the gain ratios, thereby clearing the way for their tuning to match the VROP derived gain ratios and transition of RSBAutoCal to automated mode for both dark signals and gain ratios. This solution has been fully and successfully tested with Matlab code developed to simulate RSBAutoCal gain ratio and dark signal calculations and is ready for delivery for operational implementation.

Non–small cell lung cancers (NSCLC) harboring anaplastic lymphoma kinase (ALK) gene rearrangements invariably develop resistance to the ALK tyrosine kinase inhibitor (TKI) crizotinib. Herein, we report the first preclinical evaluation of the next-generation ALK TKI, ceritinib (LDK378), in the setting of crizotinib resistance. An interrogation of in vitro and in vivo models of acquired resistance to crizotinib, including cell lines established from biopsies of patients with crizotinib-resistant NSCLC, revealed that ceritinib potently overcomes crizotinib-resistant mutations. In particular, ceritinib effectively inhibits ALK harboring L1196M, G1269A, I1171T, and S1206Y mutations, and a cocrystal structure of ceritinib bound to ALK provides structural bases for this increased potency. However, we observed that ceritinib did not overcome two crizotinib-resistant ALK mutations, G1202R and F1174C, and one of these mutations was identified in 5 of 11 biopsies from patients with acquired resistance to ceritinib. Altogether, our results demonstrate that ceritinib can overcome crizotinib resistance, consistent with clinical data showing marked efficacy of ceritinib in patients with crizotinib-resistant disease. Significance: The second-generation ALK inhibitor ceritinib can overcome several crizotinib-resistant mutations and is potent against several in vitro and in vivo laboratory models of acquired resistance to crizotinib. These findings provide the molecular basis for the marked clinical activity of ceritinib in patients with ALK-positive NSCLC with crizotinib-resistant disease. Cancer Discov; 4(6); 662–73. ©2014 AACR. See related commentary by Ramalingam and Khuri, p. 634 This article is highlighted in the In This Issue feature, p. 621

Multisite longitudinal neuroimaging designs are used to identify differential brain structural change associated with onset or progression of disease. The reliability of neuroanatomical measurements over time and across sites is a crucial aspect of power in such studies. Prior work has found that while within-site reliabilities of neuroanatomical measurements are excellent, between-site reliability is generally more modest. Factors that may increase between-site reliability include standardization of scanner platform and sequence parameters and correction for between-scanner variations in gradient nonlinearities. Factors that may improve both between- and within-site reliability include use of registration algorithms that account for individual differences in cortical patterning and shape. In this study 8 healthy volunteers were scanned twice on successive days at 8 sites participating in the North American Prodrome Longitudinal Study (NAPLS). All sites employed 3 Tesla scanners and standardized acquisition parameters. Site accounted for 2 to 30% of the total variance in neuroanatomical measurements. However, site-related variations were trivial (

The synthesis, preclinical profile, and in vivo efficacy in rat xenograft models of the novel and selective anaplastic lymphoma kinase inhibitor 15b (LDK378) are described. In this initial report, preliminary structure–activity relationships (SARs) are described as well as the rational design strategy employed to overcome the development deficiencies of the first generation ALK inhibitor 4 (TAE684). Compound 15b is currently in phase 1 and phase 2 clinical trials with substantial antitumor activity being observed in ALK-positive cancer patients.

Establishing an effective humoral immunity is an important host defense mechanism in intestinal mucosa. T follicular helper (Tfh) cells are a spectrum of CXCR5 expressing T helper cells that induce antigen-specific B cell differentiation. Because the differentiation of T helper cells is largely regulated by innate immunity, we addressed whether TRIF signaling regulates Tfh cell differentiation and its ability to trigger humoral immune responses in the intestine.CD4At baseline, CD4Our results reveal a novel role of TRIF in the regulation of humoral immunity in the intestine that can be utilized as a basis for a unique vaccine strategy.

Objective: Establishing an effective humoral immunity is an important host defense mechanism in intestinal mucosa. T follicular helper (Tfh) cells are a spectrum of CXCR5 expressing T helper cells that induce antigenspecific B cell differentiation. Because the differentiation of T helper cells is largely regulated by innate immunity, we addressed whether TRIF signaling regulates Tfh cell differentiation and its ability to trigger humoral immune responses in the intestine. Method: CD4+CXCR5+ T cells, B cells, and plasma cells in the Peyer’s patches (PPs) of WT and TRIF-deficient (TrifLPS2) mice were analyzed by flow cytometry at the baseline, 9 days post primary infection, and 7 days postsecondary infection with Y. enterocolitica. Y. enterocolitica-specific CD4+CXCR5+ T cells were generated in vitro by co-culturing peritoneal macrophages with splenic naive T cells in the presence of Y. enterocolitica lysate. WT and TrifLPS2 mice received CD4+CXCR5+ T cells isolated either from Y. enterocolitica-primed WT mice or generated in vitro. These mice were infected with Y. enterocolitica and followed up to 4 weeks. Y. enterocolitica-specific IgA and IgG were measured in stool and serum samples, respectively. Results: At baseline, CD4+CXCR5+ T cell proportion was higher but the proportion of B cells and plasma cells was lower in the PPs of TrifLPS2 mice compared to WT mice. After infection, the proportion of plasma cells also became higher in the PPs of TrifLPS2 mice compared to WT mice. Corresponding increase of Y. enterocolitica-specific stool IgA but not serum IgG was found in TrifLPS2 mice compared to WT mice. Both in vivo isolated and in vitro generated CD4+CXCR5+ T cells induced protective immunity against Y. enterocolitica infection. Conclusion: Our results reveal a novel role of TRIF in the regulation of humoral immunity in the intestine that can be utilized as a basis for a unique vaccine strategy.

Via influencing brain plasticity, aerobic exercise could contribute to the treatment of schizophrenia patients. As previously shown, physical exercise increases hippocampus volume and improves short-term memory. We now investigated gray matter density and brain surface expansion in this sample using MRI-based cortical pattern matching methods. Comparing schizophrenia patients to healthy controls before and after 3 months of aerobic exercise training (cycling) plus patients playing table football yielded gray matter density increases in the right frontal and occipital cortex merely in healthy controls. However, respective exercise effects might be attenuated in chronic schizophrenia, which should be verified in a larger sample.

Yeh P, Ezzelarab M, Bovin N, Hara H, Long C, Tomiyama K, Sun F, Ayares D, Awwad M, Cooper DKC. Investigation of potential carbohydrate antigen targets for human and baboon antibodies. Xenotransplantation 2010; 17: 197–206. © 2010 John Wiley & Sons A/S. Abstract: Background: The continued presence of a primate antibody-mediated response to cells and organs from α1,3-galactosyltransferase gene-knockout (GTKO) pigs indicates that there may be antigens other than Galα1,3Gal (αGal) against which primates have xenoreactive antibodies. Human and baboon sera were tested for reactivity against a panel of saccharides that might be potential antigen targets for natural anti-non-αGal antibodies. Methods: Human sera (n = 16) and baboon sera (n = 15) of all ABO blood types were tested using an enzyme-linked immunoadsorbent assay for binding of IgM and IgG to a panel of synthetic polyacrylamide-linked saccharides (n = 15). Human sera were also tested after adsorption on αGal immunoaffinity beads. Sera from healthy wild-type (WT, n = 6) and GTKO (n = 6) pigs and from baboons (n = 4) sensitized to GTKO pig organ or artery transplants (of blood type O) were also tested. Forssman antigen expression on baboon and pig tissues was investigated by immunohistochemistry. Results: Both human and baboon sera showed high IgM and IgG binding to αGal saccharides, α-lactosamine, and Forssman disaccharide. Human sera also demonstrated modest binding to N-glycolylneuraminic acid (Neu5Gc). When human sera were adsorbed on αGal oligosaccharides, there was a reduction in binding to αGal and α-lactosamine, but not to Forssman. WT and GTKO pig sera showed high binding to Forssman, and GTKO pig sera showed high binding to αGal saccharides. Baboon sera sensitized to GTKO pigs showed no significant increased binding to any specific saccharide. Staining for Forssman was negative on baboon and pig tissues. Conclusions: We were unable to identify definitively any saccharides from the selected panel that may be targets for primate anti-non-αGal antibodies. The high level of anti-Forssman antibodies in humans, baboons, and pigs, and the absence of Forssman expression on pig tissues, suggest that the Forssman antigen does not play a role in the primate immune response to pigs.

The Suomi National Polar Orbiting Partnership (S-NPP) Visible Infrared Imaging Radiometer Suite (VIIRS) employs a large number of temperature and voltage sensors (telemetry points) to monitor instrument health and performance. We have collected data and built tools to study telemetry and calibration parameters trends. The telemetry points are organized into groups based on locations and functionalities. Examples of the groups are: telescope motor, focal plane array (FPA), scan cavity bulkhead, radiators, solar diffuser and Solar Diffuser Stability Monitor (SDSM). We have performed daily monitoring and long-term trending studies. Daily monitoring processes are automated with alarms built into the software to indicate if pre-defined limits are exceeded. Long-term trending studies focus on instrument performance and sensitivities of Sensor Data Record (SDR) products and calibration look-up tables (LUTs) to instrument temperature and voltage variations. VIIRS uses a DC Restore (DCR) process to periodically correct the analog offsets of each detector of each spectral band to ensure that the FPA output signals are always within the dynamic range of the Analog to Digital Converter (ADC). The offset values are updated based on observations of the On-Board Calibrator Blackbody source. We have performed a long-term trend study of DCR offsets and calibration parameters to explore connections of the DCR offsets with onboard calibrators. The study also shows how the instrument and calibration parameters respond to the VIIRS Petulant Mode, spacecraft (SC) anomalies and flight software (FSW) updates. We have also shown that trending studies of telemetry and calibration parameters may help to improve the instrument calibration processes and SDR Quality Flags.

Anaplastic lymphoma kinase (ALK) is a receptor tyrosine kinase belonging to the insulin receptor superfamily. Expression of ALK in normal human tissues is only found in a subset of neural cells, however it is involved in the genesis of several cancers through genetic aberrations involving translocation of the kinase domain with multiple fusion partners (e.g., NPM-ALK in anaplastic large cell lymphoma ALCL or EML4-ALK in non-small cell lung cancer) or activating mutations in the full-length receptor resulting in ligand-independent constitutive activation (e.g., neuroblastoma). Here we are reporting the discovery of novel and selective anaplastic lymphoma kinase inhibitors from specific modifications of the 2,4-diaminopyridine core present in TAE684 and LDK378. Synthesis, structure activity relationships (SAR), absorption, distribution, metabolism, and excretion (ADME) profile, and in vivo efficacy in a mouse xenograft model of anaplastic large cell lymphoma are described.

Sustainable water management decisions are often made on the support of water quantity and quality models with a focus on prediction uncertainty. Unfortunately, limited observational data severely constrains the design of accurate water models for these decisions. To address this challenge, this paper presents our initial efforts to deploy STORM 3 data loggers from WaterLOG to collect real-time and in-situ data at fine temporal granularities to monitor the pond at Lamar University in Beaumont, TX. We present the details about how to set up STORM 3, integrate H-377 water temperature sensor probe from WaterLOG, and validate collected water temperature data. Since July 2014, our deployed STORM 3 is collecting water temperature data once every 15 minutes. On-going research is underway to revise the deployment of our STORM 3, and to provide water temperature data for decision making on water sustainability management.

Objective: This study aimed to characterize the effects of mirtazapine on polysomnographic sleep, especially slow wave sleep (SWS) and rapid eye movement (REM) sleep, as well as its effects on clinical symptoms in patients with major depressive disorder (MDD). Method: Sixteen MDD patients were treated with mirtazapine 30 mg taken 30 minutes before bedtime. Polysomnographic and subjective sleep, as well as other clinical data, were collected at baseline and on Days or Nights 2, 9, 16, 30, and 58 during treatment. We used repeated measures analysis of variance, including pairwise comparison, to analyze data statistically. Results: Mirtazapine administration increased total SWS and the SWS in the first sleep cycle, but not SWS in the second sleep cycle. The medication increased REM latency and the duration of the first REM episode; it also decreased the number of REM episodes. Simultaneously, mirtazapine significantly reduced wake-after-sleep onset and scores on the Athens Insomnia Scale. After patients took the medication, scores on the Hamilton Depression Rating Scale-17 (HDRS-17) decreased rapidly and continuously. The changes on the Beck Depression Inventory-II were consistent with those on the HDRS-17. The medication has a tendency to increase weight. Conclusions: Mirtazapine significantly improved sleep quality, reversed sleep markers of depression, and reduced depressive symptoms in this group of MDD patients.

Modern assisted history matching tools allow an engineer to specify the uncertain parameters in a simulation dataset then perform an optimization to minimize the difference between observed field data and the corresponding simulation outputs. During this optimization, multiple simulations are run, with the uncertain parameters varying within the ranges specified by the engineer. The difference between the observed field data and the simulation output is measured by an objective function. Standard objective functions have been reported in the literature for the difference between observed and simulated production and injection rates, and for measurements in time and space done at observation wells. In this work we incorporate an additional objective function term that measures the difference between the observed and simulated steam chamber location and shape. In addition, the differences in 3D volumes were visualized, which lead to a better physical understanding of what parameters should be adjusted during a history match.For a steam injection process like Steam Assisted Gravity Drainage (SAGD), 4D seismic may be used to determine where a steam chamber is located in a reservoir at a point in time. The objective function developed in this work measures the difference between an observed chamber's shape and location in the reservoir and the corresponding shape and location for the chamber indicated in the simulation output. The objective function is a binary mismatch function, checking each simulation grid block to see if the seismic chamber and the simulation chamber agree or disagree, and calculating the ratio of the total volume of disagreements over the total volume of agreements.This steam chamber mismatch function was included in an assisted history match performed on a well pair from Suncor Energy's SAGD project at Firebag. The inclusion of this additional information added additional constraints to the simulation model, leading to a conceptually more dependable history match and a better geological and dynamic characterization of the reservoir.

The synthesis, preclinical profile, and in vivo efficacy in rat xenograft models of the novel and selective anaplastic lymphoma kinase inhibitor 15b (LDK378) are described. In this initial report, preliminary structure-activity relationships (SARs) are described as well as the rational design strategy employed to overcome the development deficiencies of the first generation ALK inhibitor 4 (TAE684). Compound 15b is currently in phase 1 and phase 2 clinical trials with substantial antitumor activity being observed in ALK-positive cancer patients.

Lactobacillus johnsonii (Ljo) N6.2 has been shown to mitigate the development of type 1 diabetes when administered to diabetes-prone rats. The specific mechanisms underlying this observed response remain under investigation. The objective of this study was to assess the effect of Ljo N6.2 on mucosal inflammatory response using differentiated Caco-2 monolayers. The mRNA expression levels of CCL20, CXCL8, and CXCL10 chemokines were determined by qRT-PCR. Ljo at 10 11 CFU/L induced a strong response in all chemokines examined. To assess the specific host-signaling pathways involved, we performed RT-PCR amplification of Toll-like receptors (TLR) and nucleotide-binding oligomerization domainlike receptors. TLR7 and TLR9 expression levels were induced 4.2- and 9-fold, respectively, whereas other TLR and nucleotide-binding oligomerization domain receptors were not modified. A similar effect was observed in Caco-2 monolayers treated with Ljo cell-free extract or purified nucleic acids (NA). Increased levels of IFN type 1 and IFN regulators Stat1 and IRF7 followed the upregulation of TLR9. Activation of TLR9 was also evidenced by increased Frizzled 5 expression in Ljo-treated Caco-2 cells and an increase in the number of Paneth cells in Ljo‐fed, diabetes-prone rats. These results are in agreement with the polarizing-tolerizing mechanism recently described in which the apical stimulation of TLR9 in intestinal epithelial cells leads to a higher state of immunologic alertness. Furthermore, these results suggest that live probiotics could be, in the future, replaced with select cellular components. J. Nutr. doi: 10.3945/jn.110.135517.

The continued presence of a primate antibody-mediated response to cells and organs from alpha1,3-galactosyltransferase gene-knockout (GTKO) pigs indicates that there may be antigens other than Gal alpha 1,3Gal (alpha Gal) against which primates have xenoreactive antibodies. Human and baboon sera were tested for reactivity against a panel of saccharides that might be potential antigen targets for natural anti-non-alpha Gal antibodies.Human sera (n = 16) and baboon sera (n = 15) of all ABO blood types were tested using an enzyme-linked immunoadsorbent assay for binding of IgM and IgG to a panel of synthetic polyacrylamide-linked saccharides (n = 15). Human sera were also tested after adsorption on alpha Gal immunoaffinity beads. Sera from healthy wild-type (WT, n = 6) and GTKO (n = 6) pigs and from baboons (n = 4) sensitized to GTKO pig organ or artery transplants (of blood type O) were also tested. Forssman antigen expression on baboon and pig tissues was investigated by immunohistochemistry.Both human and baboon sera showed high IgM and IgG binding to alpha Gal saccharides, alpha-lactosamine, and Forssman disaccharide. Human sera also demonstrated modest binding to N-glycolylneuraminic acid (Neu5Gc). When human sera were adsorbed on alpha Gal oligosaccharides, there was a reduction in binding to alpha Gal and alpha-lactosamine, but not to Forssman. WT and GTKO pig sera showed high binding to Forssman, and GTKO pig sera showed high binding to alpha Gal saccharides. Baboon sera sensitized to GTKO pigs showed no significant increased binding to any specific saccharide. Staining for Forssman was negative on baboon and pig tissues.We were unable to identify definitively any saccharides from the selected panel that may be targets for primate anti-non-alpha Gal antibodies. The high level of anti-Forssman antibodies in humans, baboons, and pigs, and the absence of Forssman expression on pig tissues, suggest that the Forssman antigen does not play a role in the primate immune response to pigs.

Detailed measurements of mean and turbulent velocity components and turbulent shear stress components have been made for jet flows of 20-, 40-, and 60-deg swirlers for 750 and 1500 mm H{sub 2}O supply air pressure conditions. When the mean velocity components are normalized based on the maximum axial velocity at the nozzle exit, there is a very close similarity between normalized profiles at different supply pressures for each swirler. The recirculation zone, which depends on the degree of swirl and the swirler geometry, is not well established near the nozzle exit for the case of high ratio of the inner to outer radius of the swirler and of low degree of swirl. This can create a serious deposition problem of fuel droplets onto the hub surface. Measurements of turbulent velocity components demonstrate that each jet can be considered to be close to local isotropic turbulence except at the nozzle exit, where the magnitudes and decay of the fluctuations depend strongly on flow direction. 14 refs.

Introduction The purpose of the study was to evaluate a recently introduced aesthetic restorative system in respect of: • Presentation and organisation of the package. • Its handling characteristics. • Its polishability. • Shade selection procedures and aesthetic outcome. Material and method The material, Esthet.X Micro Matrix system (Dents-ply), was evaluated by general dental practitioners from a clinical research group known as PEER (Product and Equipment Evaluation and Research). The evaluators were asked to use the material, following manufacturer's instructions, in a variety of cases. A questionnaire was designed specifically for this study and the questions were directed to both the dentists and their dental nurses. The results were analysed statistically and represented in visual analogue scales. Result The material was found to be useful and generally well received. Its use was thought to be technique sensitive but good to excellent results can be achieved. Areas where the evaluators thought the product could be improved were also discussed.

Non-small cell lung cancers (NSCLC) harboring anaplastic lymphoma kinase (ALK) gene rearrangements are sensitive to the ALK tyrosine kinase inhibitor (TKI) crizotinib. However, these cancers invariably relapse due to the development of resistance, and approximately 1/3 of such cancers develop resistance mutations within the ALK tyrosine kinase domain. Here we report the preclinical evaluation of the next-generation ALK TKI, LDK378 in the setting of crizotinib resistance. Using EML4-ALK mutant Ba/F3 cellular models, in vivo models of acquired resistance to crizotinib, and novel cell lines established from biopsies of crizotinib-resistant NSCLC patients, we have examined the efficacy of LDK378 in crizotinib-naïve and -resistant ALK-positive cancers. These studies reveal that LDK378 is more potent than crizotinib and effectively overcomes resistance in vitro and in vivo. In particular, LDK378 inhibits ALK harboring crizotinib resistance mutations, including L1196M, G1269A, I1171T and S1206Y. Cell lines derived from crizotinib-resistant biopsies were sensitive to LDK378, including one that did not harbor an ALK resistance mutation and was also sensitive to crizotinib, suggesting that some crizotinib-resistant cancers with wildtype ALK are still sensitive to complete ALK inhibition. We observed that LDK378 did not effectively overcome two crizotinib-resistant ALK mutations, G1202R and F1174C ALK, and mutations in one of these residues was identified in 5 out of 11 biopsies from patients with acquired resistance to LDK378. Altogether our results demonstrate that LDK378 can overcome many mechanisms of crizotinib resistance, consistent with emerging clinical data showing marked efficacy of LDK378 in patients with crizotinib-resistant disease. Citation Format: Luc Friboulet, Nanxin Li, Ryohei Katayama, Christian C. Lee, Justin F. Gainor, Adam S. Crystal, Pierre-Yves Michellys, Mark M. Awad, Noriko Yanagitani, Sungjoon Kim, AnneMarie Pferdekamper, Jie Li, Shailaja Kasibhatla, Frank Sun, Xiuying Sun, Su Hua, Peter McNamara, Sidra Mahmood, Elizabeth L. Lockerman, Naoya Fujita, Makoto Nishio, Jennifer L. Harris, Alice T. Shaw, Jeffrey A. Engelman. The ALK inhibitor LDK378 overcomes crizotinib resistance in non-small cell lung cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 957. doi:10.1158/1538-7445.AM2014-957

Selective RAF inhibitors have significant activity in patients with metastatic melanoma whose tumors express BRAFV600E. However, not all patients respond equally well to treatment and the duration of response is often limited to less than 6 months. LGX818 was developed with the hypothesis that a more potent inhibitor with excellent pharmacological properties would maximize the degree and duration of patient response. LGX818 is a highly potent RAF inhibitor with selective anti-proliferative and apoptotic activity in cells expressing BRAFV600E. In the A375 (BRAFV600E) human melanoma cell line LGX818 suppresses phospho-ERK (EC50 = 3 nM) leading to potent inhibition of proliferation (EC50 = 4 nM). No significant activity was observed against a panel of 100 kinases (IC50 > 900 nM) and LGX818 did not inhibit proliferation of > 400 cell lines expressing wild-type BRAF. Contributing to the high potency of LGX818 is the extremely slow off-rate from BRAFV600E which is not observed with other RAF inhibitors. In biochemical assays the dissociation half-life was >24 hours which translated into sustained target inhibition in cells following drug wash-out. Single dose PK/PD studies in human melanoma xenograft models (BRAFV600E) indicated that LGX818 treatment at oral doses as low as 6 mg/kg resulted in strong (75%) and sustained (>24 hours) decrease in phospho-MEK, even following clearance of drug from circulation. Decreases in phospho-ERK were consistent with phospho-MEK but markers of downstream transcriptional output (DUSP6 and SPRY4) appeared to provide a more sensitive measure of pathway activation. LGX818 induced tumor regression in multiple BRAF mutant human tumor xenograft models grown in immune compromised mice and rats at doses as low as 1 mg/kg. Consistent with the in vitro data, LGX818 was inactive against BRAF wild-type tumors at doses up to 300 mg/kg bid, with good tolerability and linear increase in exposure. Efficacy was also achieved in a more disease-relevant spontaneous metastatic melanoma and a model of melanoma brain metastasis. LGX818 is a potent and selective RAF kinase inhibitor with unique biochemical properties that contribute to an excellent pharmacological profile. A Phase I clinical trial in patients with BRAF mutant tumors is ongoing. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3790. doi:1538-7445.AM2012-3790

A c-MET/ALK kinase inhibitor crizotinib has demonstrated significant activity in lung cancer patients with EML4-ALK in clinical studies. However relapse (or acquired resistance) has also been reported. We have developed crizotinib resistant tumor models and used the models to evaluate the ALK inhibitor LDK378. LDK378 is a potent and selective ALK inhibitor that does not cross react with c-MET. In a mouse xenograft tumor model derived from the EML4-ALK+ lung cancer cell line NCI-H2228, LDK378 caused complete tumor regression when dosed orally at 25 mg/kg/day. After tumor bearing animals had been treated with LDK378 at 50 mg/kg/day for 14 days, remission was maintained for more than 4 months. In several NCI-H2228 tumor models that were induced to become resistant to crizotinib, LDK378 demonstrated efficacy at 50 mg/kg/day. Based on 4-wk GLP toxicology studies the drug exposure associated with this dose is predicted to be below the exposure at the MTD in humans. ALK resistance mutations reported in crizotinib relapsed patients were also found in the crizotinib resistant NCI-H2228 tumor models. The results from these preclinical studies suggest that LDK378 may be active in crizotinib-relapsed patients. A phase I clinical study of LDK378 has recently begun in both crizotinib-relapsed and crizotinib-naïve patients. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2011 Nov 12-16; San Francisco, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2011;10(11 Suppl):Abstract nr B232.

We recently reported a new class of allosteric inhibitors, exemplified by GNF-2, that selectively inhibit the proliferation of Bcr-Abl dependent cells. Here we demonstrate, using selection for resistant Bcr-Abl clones, site-directed mutagenesis, affinity chromatography, and steady-state kinetics, that GNF-2 inhibits Bcr-Abl kinase activity by binding to the Abl myristate binding pocket and stabilizes the auto-inhibited conformation. We demonstrate that the 2-hydroxyethyl amide analog of GNF-2, GNF-5, in combination with ATP-competitive inhibitors such as nilotinib and dasatinib can overcome the T315I “gatekeeper” mutant of Bcr-Abl which is resistant to all clinically approved Bcr-Abl inhibitors. These studies demonstrate that targeting of the Abl myristate binding site can provide an important pharmacological means to overcome mutations that cause resistance to ATP-competitive inhibitors. Citation Information: Mol Cancer Ther 2009;8(12 Suppl):B165.

An examination of colour term saliency among children is provided in this paper. This discussion is organized around the notions of colour term development noted in the 1iterature and with experiments conducted by researchers in the distinction of colours by children. This review suggests that colour recognition, colour naming and that focal colours are cognitively recognized before hue colours, as a result of their saliency with respect to the colour spectrum.

The role of PGs in the maintenance and the control of fetal and placental hemodynamics was suggested. Umbilical vessels having smooth muscle that is not innervated may be particularly responsive to vasoactive agents such as PGs. Six UA were freshly obtained from normal term pregnancies. Biosynthesis of PGE2, PGF2α, thromboxane A2 (analyzed as TxB2) and prostacyclin (PGI2) (Measured as 6-keto PGF1α) was studied by incubating UA microsomes with (14C) arachidonic acid (AA) and with (14C) cyclic PG endoperoxide (PGH2), separation by thin layer chromatography and scintillation counting. Percent recovery of (14C) AA and (14C) PGH2 as PGs was 1.73 and 36.4 respectively. The relative percent PGs recovered was (x ± SEM): In addition, PGI2 synthesis by UA microsomes was determined by the degree of inhibition of platelet aggregation compared with inhibition by known PGI2 standards and was 7μg/mg protein. Azo analog I inhibited PGs biosynthesis in duplicate incubations. The results suggest that PGs synthesis by the vascular wall of the UA may contribute to the regulation of vascular tone and platelet vessel wall interaction, thus, modulating fetoplacental blood flow. PGs inhibitors may alter balanced homeostasis in the fetus and the placenta.