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PDF file, 181K, Supplementary Figure S3 Sustained inhibition of Akt and Erk phosphorylation by combining WZ4002 and E7050.

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PDF file, 115K, Supplementary Figure S2 E7050 inhibits tumor cell motility induced by HGF overexpression.

PDF file, 138K, Supplementary Figure S6 Treatment with WZ4002 and/or E7050 does not induce apoptosis in vivo.

PDF file, 423K, Supplementary Figure S4 Cytostatic effects of WZ4002 and E7050 treatment.

PDF file, 315K, Supplementary Figure S5 Relative body weight of mice during treatment with WZ4002 and/or E7050.

PDF file, 40K, Supplementary Table S1 IC50 values of each compound in MTT assay.

Although the EGF receptor tyrosine kinase inhibitors (EGFR-TKI) erlotinib and gefitinib have shown dramatic effects against EGFR mutant lung cancer, patients become resistant by various mechanisms, including gatekeeper EGFR-T790M mutation, Met amplification, and HGF overexpression, thereafter relapsing. Thus, it is urgent to develop novel agents to overcome EGFR-TKI resistance. We have tested the effects of the mutant-selective EGFR-TKI WZ4002 and the mutant-selective Met-TKI E7050 on 3 EGFR mutant lung cancer cell lines resistant to erlotinib by different mechanisms: PC-9/HGF cells with an exon 19 deletion, H1975 with an L858R mutation, and HCC827ER with an exon 19 deletion, with acquired resistance to erlotinib because of HGF gene transfection, gatekeeper T790M mutation, and Met amplification, respectively. WZ4002 inhibited the growth of H1975 cells with a gatekeeper T790M mutation, but did not inhibit the growth of HCC827ER and PC-9/HGF cells. HGF triggered the resistance of H1975 cells to WZ4002, whereas E7050 sensitized HCC827ER, PC-9/HGF, and HGF-treated H1975 cells to WZ4002, inhibiting EGFR and Met phosphorylation and their downstream molecules. Combined treatment potently inhibited the growth of tumors induced in severe-combined immunodeficient mice by H1975, HCC827ER, and PC-9/HGF cells, without any marked adverse events. These therapeutic effects were associated with the inhibition of EGFR and Met phosphorylation in vivo. The combination of a mutant-selective EGFR-TKI and a Met-TKI was effective in suppressing the growth of erlotinib-resistant tumors caused by gatekeeper T790M mutation, Met amplification, and HGF overexpression. Further evaluations in clinical trials are warranted. Mol Cancer Ther; 11(10); 2149–57. ©2012 AACR.

Direct observation of valence electron density reflecting orbital degrees of freedom on partially filled degenerate orbitals.

PDF file - 785K, Supplementary Figure 1. Expression of total BIM and ratio of BIM isoforms in NSCLC cell lines. Supplementary Figure 2. Flow cytometric analysis showing that EGFR mutated cell lines carrying the BIM polymorphism have low susceptibility to gefitinib-induced apoptosis. Supplementary Figure 3. Vorinostat-induced up-regulation of BIM enhances apoptosis in the HCC2279 cell line with the BIM polymorphism.Supplementary Figure 4. Knockdown or overexpression of BIM affects activation of caspase-3/7 induced by gefitinib and vorinostat in EGFR mutant cells with BIM polymorphism. Supplementary Figure 5. Detection of BIM polymorphism in human peripheral blood mononuclear cells (PBMC) and serum. Supplementary Table 1. Status of EGFR mutation and BIM polymorphism in NSCLC cell lines.

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BIM (BCL2L11) is a BH3-only proapoptotic member of the Bcl-2 protein family. BIM upregulation is required for apoptosis induction by EGF receptor (EGFR) tyrosine kinase inhibitors (EGFR-TKI) in EGFR-mutant forms of non–small cell lung cancer (NSCLC). Notably, a BIM deletion polymorphism occurs naturally in 12.9% of East Asian individuals, impairing the generation of the proapoptotic isoform required for the EGFR-TKIs gefitinib and erlotinib and therefore conferring an inherent drug-resistant phenotype. Indeed, patients with NSCLC, who harbored this host BIM polymorphism, exhibited significantly inferior responses to EGFR-TKI treatment than individuals lacking this polymorphism. In an attempt to correct this response defect in the resistant group, we investigated whether the histone deacetylase (HDAC) inhibitor vorinostat could circumvent EGFR-TKI resistance in EGFR-mutant NSCLC cell lines that also harbored the BIM polymorphism. Consistent with our clinical observations, we found that such cells were much less sensitive to gefitinib-induced apoptosis than EGFR-mutant cells, which did not harbor the polymorphism. Notably, vorinostat increased expression in a dose-dependent manner of the proapoptotic BH3 domain-containing isoform of BIM, which was sufficient to restore gefitinib death sensitivity in the EGFR mutant, EGFR-TKI–resistant cells. In xenograft models, while gefitinib induced marked regression via apoptosis of tumors without the BIM polymorphism, its combination with vorinostat was needed to induce marked regression of tumors with the BIM polymorphism in the same manner. Together, our results show how HDAC inhibition can epigenetically restore BIM function and death sensitivity of EGFR-TKI in cases of EGFR-mutant NSCLC where resistance to EGFR-TKI is associated with a common BIM polymorphism. Cancer Res; 73(8); 2428–34. ©2013 AACR.

Background rates in several pixel array detectors are investigated with an eye toward using them with hard, >6 keV, x-rays in very low-rate experiments - e.g. at signal rates, 15 Pages, 12 Figures, v2 ads link and e-mail

Physical function impairment in patients with low back pain (LBP) occurs due to the influence of psychosocial factors. Only a few studies have objectively evaluated physical function. We aimed to objectively assess the physical functions of individuals subjects with LBP, and clarify the association between physical function and psychosocial factors. We enrolled 411 individuals with LBP working in special needs schools. We examined their degree of pain, and the psychosocial factors strength through the STarT Back Tool, which categorized them into the low-risk, medium-risk, and high-risk groups. We assessed their abdominal muscle endurance, lower limb muscle strength, and hip joint flexibility. The relationships between these physical functions and psychosocial factors were analyzed by logistic regression models. Those in the high-risk group had significantly lower abdominal muscle and lower limb muscle strength (p

Although tyrosine kinase inhibitors markedly improve the clinical outcome of chronic myeloid leukemia (CML), blast crisis in CML (CML-BC) still has a poor prognosis. Many chromosomal abnormalities have been reported in CML-BC and may contribute to therapeutic resistance, disease progression, and prognosis. Herein, we report a rare chromosome abnormality with der(16)t(1;16)(q12;q11.2) in CML-BC. It has been demonstrated that this chromosomal abnormality is associated with disease progression and poor prognosis in other malignancies, such as Ewing sarcoma. A 70-year-old man with CML who had been treated with imatinib and dasatinib was admitted to our hospital after complaining for several weeks of fatigue and dyspnea and diagnosed with CML-BC. His tumor cells presented additional chromosomal abnormality with der(16)t(1;16)(q12;q11.2), which has never been reported in CML cases. We successfully treated him using cytotoxic agents combined with ponatinib, and this chromosome abnormality was detected via G-banding. Our patient has lived for over 8 months without any progression with ponatinib treatment alone. Although the biological function of this chromosomal abnormality remains unclear, the satellite DNA of 1q12, which induces genomic instability in other malignancies, and the loss of 16q may contribute to the disease progression of CML in this case. In conclusion, this paper is the first to report on the case of CML-BC with der(16)t(1;16)(q12;q11.2).

Troponin (Tn) is composed of three subunits (TnI, TnC and TnT) that bind Ca2+ and regulate striated muscle contraction in vertebrates. TnT’s function has been extensively described in vertebrates, but its role has been obscure in molluscan muscles. Our previous work indicated that the TnC and TnI subunits work in adductor phasic muscle, but not in catch muscle. Here, we have characterized TnT from the Japanese bivalve pearl oyster Pinctada fucata to start to explain the function of Tn in molluscan muscle contraction. We determined the primary structure of the full-length TnT protein from the P. fucata adductor muscle (Pifuc-TnT), and found that it is composed of 316 amino acid residues with a predicted molecular mass of 37.4 kDa. Multiple sequence alignment showed that Pifuc-TnT has an extension of >60 residues at the C-terminus that are not present in vertebrate TnTs, including known TnTs from other mollusks. Pifuc-TnT gene structure predictions using Splign alignment of the cDNA generated in this study and genome sequences indicated that Pifuc-TnT consists of 13 exons. Start and stop codons are located in exons 2 and 12, respectively. Quantitative real-time PCR revealed that the Pifuc-TnT gene was predominantly expressed in adductor phasic muscle, weakly in adductor catch muscle, slightly in gill, and not at all in mantle and foot. These findings suggest that TnT plays a regulatory role in adductor phasic muscle contraction, but not in catch contraction. Isothermal titration calorimetry revealed that unlike vertebrate TnTs, Pifuc-TnT does not interact with P. fucata tropomyosin-1 nor with tropomyosin-2. These findings in P. fucata imply that Tn functions differently in molluscan muscle than it does in vertebrates.

The identification of acquired resistance mutations has been essential in non-small-cell lung cancer (NSCLC) patients with epidermal growth factor receptor (EGFR) active mutations. Rebiopsy plays a pivotal role in selecting the optimal treatment for patients who develop resistance to initial EGFR-tyrosine kinase inhibitors (EGFR-TKIs). This multicenter, observational study was conducted to investigate the details of rebiopsy in Japanese clinical practice. The primary endpoints were the implementation rate of rebiopsy and the concordance rate for T790M mutation detection between histological and cytological specimens using the cobas EGFR Mutation Test, version 2. One hundred ninety-four patients with EGFR-mutant NSCLC were enrolled, and 120 patients developed acquired resistance to EGFR-TKIs. The median age was 68 years (range 20–87), and 52.5% of the patients were women. Rebiopsy was performed in 109 patients, and the implementation rate of rebiopsy was 90.8%. The success rates of rebiopsy in the total, histology, cytology and liquid biopsy populations were 67.9%, 81.3%, 66.7% and 43.8%, respectively. The positive percent agreement and the negative percent agreement in the detection of the T790M mutation between the histological and cytological specimens were both 90.9%. Obtaining histological or cytological tissue samples at rebiopsy may contribute to improving the detection rate of the T790M mutation (trial registration number: UMIN000026019).

Anaplastic lymphoma kinase (ALK)-positive lung cancer is a rare cancer that occurs in approximately 5% of non-small-cell lung cancer (NSCLCs) patients. Despite the excellent efficacy of ALK-tyrosine kinase inhibitor in ALK-positive NSCLCs, most patients experience resistance. We conducted a phase II study to investigate the combination of alectinib with bevacizumab in ALK-positive NSCLC patients after failure of alectinib. In this study, ALK-positive nonsquamous NSCLC patients previously treated with alectinib received bevacizumab 15 mg/kg on day 1 every 3 weeks and alectinib 600 mg/day until disease progression. The primary endpoints were progression-free survival (PFS) and the safety of alectinib and bevacizumab. The secondary endpoints included overall survival (OS) and correlation of circulating tumor DNA and plasma proteins with PFS. Of the 12 patients treated, the median PFS was 3.1 months (95% CI 1.2–16.1), and the median OS was 24.1 months (95% CI 8.3-not estimable). The EML4-ALK fusion gene in circulating tumor DNA was significantly correlated with shorter PFS (1.2 months vs. 11.4 months, HR 5.2, p = 0.0153). Two patients experienced grade 3 adverse events; however, none of the patients required dose reduction. Although the primary endpoint was not met, alectinib combined with bevacizumab showed clinical efficacy in ALK-positive patients.

Inelastic x-ray scattering spectra of four organic liquids, n-hexane, cyclohexane, ethylene glycol dimethyl ether, and 1,4-dioxane, were measured, and the sound velocity in the nm−1 wavenumber and meV energy regimes was determined. Compared with the corresponding values in the hydrodynamic limit, the sound velocity in the nm−1 regime was faster, and the positive dispersion of the longitudinal modulus was stronger in liquids composed of ring structures (cyclohexane and 1,4-dioxane) than in those of linear chain structures ( n-hexane and ethylene glycol dimethyl ether). Molecular dynamics simulation of n-hexane and cyclohexane was also performed. The difference in the positive dispersion of the longitudinal modulus was reproduced by simulation, and it was elucidated by the difference in the longitudinal modulus in the q = 0 limit and the THz frequency regime. The excess part of the longitudinal modulus from the hydrodynamic limit was further divided into various contributions, and the smaller excess modulus of n-hexane was mainly ascribed to two reasons. The first one is that the shear modulus of n-hexane is smaller in the THz regime, and the second one is that the positive dispersion of the bulk modulus due to the vibrational energy relaxation is weaker. The second mechanism was further interpreted in terms of the fast vibrational energy relaxation of intramolecular modes associated with the chain deformation of n-hexane.

Highlights • A patient exhibited IgG4-related hypothalamo-hypophysitis. • Prominent high-signal areas of swelling were observed in the hypothalamus, tuber cinereum, infundibulum, and bilateral optic nerve systems. • MRI T1WI with contrast media demonstrated enhanced neurohypophysis and cystic swelling, and compressed anterior pituitary. • MRI findings improved rapidly after 4 days of steroid therapy.

The sound velocity of Fe3C was measured at pressures from 33 to 86 GPa and at ambient and high temperatures up to 2300 K using inelastic X-ray scattering (IXS) from laser-heated samples in diamond anvil cells (DACs). The compressional velocity (VP) and density of Fe3C at room temperature were observed to follow a linear relationship (Birch's law). The temperature dependency of Birch's law was not clearly observed and can be ignored. Birch's law for Fe3C is expressed by: V P = 1.09 ± 0.14 × ρ − 1.79 ± 1.26 . The result indicates that VP and VS (shear velocity) of the preliminary reference Earth model (PREM) inner core at the Inner Core Boundary (ICB) were by 12% and 48% smaller than those of Fe3C, which could be accounted for by the premelting effect by analogy from pure Fe or by partial melting of the Fe–Fe3C mixture in the inner core.

A tetranuclear iron grid-type spin crossover complex, [FeII4(HL)4](BF4)4·6H2O (1) (H2L = 2-(4-hydroxyphenyl)-4,5-bis{6-(3,5-dimethylpyrazol-1-yl)pyrid-2- yl}-1H-imidazole), was synthesized. Magnetic susceptibility measurements revealed that 1 shows gradual spin crossover behavior above 200 K. The electronic states of the compound were elucidated by Mossbauer spectra.

Despite the technological importance of supercritical fluids, controversy remains about the details of their microscopic dynamics. In this work, we study four supercritical fluid systems -- water, Si, Te, and Lennard-Jones fluid -- \emph{via} classical molecular dynamics simulations. A universal two-component behavior is observed in the intermolecular dynamics of these systems, and the changing ratio between the two components leads to a crossover from liquidlike to gaslike dynamics, most rapidly around the Widom line. We find evidence to connect the liquidlike component dominating at lower temperatures with intermolecular bonding, and the component prominent at higher temperatures with free-particle, gaslike dynamics. The ratio between the components can be used to describe important properties of the fluid, such as its self-diffusion coefficient, in the transition region. Our results provide insight into the fundamental mechanism controlling the dynamics of supercritical fluids, and highlight the role of spatiotemporally inhomogenous dynamics even in thermodynamic states where no large-scale fluctuations exist in the fluid., 26 pages, 6 figures (main) 13 pages, 7 figures (supplemental)

We compare several different ways of measuring the energy resolution for meV-resolved inelastic x-ray scattering (IXS): using scattering from poly(methyl methacrylate), PMMA, using scattering from borosilicate glass (Tempax), and using powder diffraction from aluminum. All of these methods provide a reasonable first approximation to the energy resolution, but, also, in all cases, inelastic contributions appear over some range of energy transfers. Over a range of +-15 meV energy transfer there is good agreement between the measurements of PMMA and Tempax at low temperature, and room temperature powder diffraction from aluminum so we consider this to be a good indication of the true resolution of our ~1.3 meV spectrometer. We self-consistently determine the resolution over a wider energy range using the temperature, momentum and sample dependence of the measured response. We then quantitatively investigate the inelastic contributions from the PMMA and Tempax, and their dependence on momentum transfer and temperature. The resulting data allows us to determine the resolution of our multi-analyzer array efficiently using a single scan. We demonstrate the importance of this procedure by showing that the results of the analysis of a spectrum from a glass are changed by using the properly deconvolved resolution function. We also discuss the impact of radiation damage on the scattering from PMMA and Tempax., Comment: 11 pages 10 figures - this version, 3.01, adds figure 10

We report a new experimental approach for observing mesoscopic fluctuations underlying the thermodynamic anomalies of ambient liquid water. In this approach, two sound velocity measurements with different frequencies, namely inelastic X-ray scattering (IXS) in THz band and ultrasonic (US) in MHz band, are required to investigate the relaxation phenomenon with the characteristic frequency between the two aforementioned frequencies. We performed IXS measurements to obtain the IXS sound velocity of liquid water from the ambient conditions to the supercritical region of liquid-gas phase transition (LGT) and compared the results with the US sound velocity in the literature. We found that the ratio of the two sound velocities, Sf, which corresponds to the relaxation intensity, exhibits a simple but significant change. Two distinct rises were observed in the high-temperature and low-temperature regions, implying that two relaxation phenomena exist: in the high-temperature region, a peak was observed near the LGT critical ridge line, which was linked with changes in the density fluctuation and isochoric and isobaric specific heat capacities; in the low-temperature region, Sf increased toward the low-temperature region, which was linked with the change in the isochoric heat capacity. We concluded that these two relaxation phenomena are originated from critical fluctuations of liquid-gas phase transition (LGT) and liquid-liquid phase transition, respectively. The linkage between Sf and isochoric heat capacity in the low-temperature region proves that the relaxation is the cause of the well-known heat capacity anomaly of ambient liquid water. In this study, both LGT and LLT critical fluctuations were observed, and the relationship between thermodynamics and the critical fluctuations was comprehensively discussed.