Real-space imaging of several molecular layers of C60in the rotational glass phase.

C60is a model system to study molecule-surface interactions and phase transitions due to its high symmetry and strong covalentπbonding within the molecule versus weak van-der-Waals coupling between neighboring molecules. In the solid, at room temperature, the molecule rotates and behaves as a sphere. However, the pentagonal and hexagonal atomic arrangement imposes deviations from the spherical symmetry that become important at low temperatures. The orientation of the C60can be viewed to represent classic spins. For geometrical reasons the preferred orientation of neighboring C60cannot be satisfied for all of the neighboring molecules, making C60a model for disordered spin systems with frustration. We study several molecular layers of C60islands on highly oriented pyrolytic graphite using scanning tunneling microscopy at liquid nitrogen temperatures. By imaging several layers we obtain a limited access to the three-dimensional rotational structure of the molecules in an island. We find one rotationally disordered layer between two partially rotationally ordered layers with hexagonal patterns. This exotic pattern shows an example of the local distribution of order and disorder in geometrically frustrated systems. Scanning tunneling spectroscopy data confirms the weak interactions of neighboring molecules.

Shell effects and free-electrons in electromigrated oxidized Cu-nanocontacts.

Electromigration in interconnects continues to be an important field of study in integrated circuits as the interconnects are planned to shrink in size at comparable pace as the semiconductor functional elements. Through shrinking the interconnects approach the regime where quantum size effects become important. The observation of quantum size and shell effects is usually restricted either to low-temperatures or vacuum conditions or to chemically inert materials such as Au. Here, we show that in electromigrated Cu nanocontacts such effects can be observed at room temperature and room pressure even in the presence of oxidation. Our data provide evidence that the nanocontacts are nearly spherical objects with a triangular-cylindrical symmetry of their electronic wave functions with a stronger free-electron-like character compared to previous results. We do not observe a detrimental effect of oxygen. The presence of shell effects has implications for the technological use of Cu nanocontacts as interconnects in integrated circuits and could lead to the use of electronic wave functions of shells in such interconnects.

Molecular self-assembly of DBBA on Au(111) at room temperature.

We have investigated the self-assembly of the graphene nanoribbon molecular precursor 10,10'-dibromo-9,9'-bianthryl (DBBA) on Au(111) with frequency modulation scanning force microscopy (FM-SFM) at room temperature combined with ab initio calculations. For low molecular coverages, the molecules aggregate along the substrate herringbone reconstruction main directions while remaining mobile. At intermediate coverage, two phases coexist, zigzag stripes of monomer chains and decorated herringbones. For high coverage, the molecules assemble in a dimer-striped phase. The adsorption behaviour of DBBA molecules and their interactions are discussed and compared with the results from ab initio calculations.

Pediatric Sedation and Analgesia Outside the Operating Room: Combining Intranasal Fentanyl and Inhaled Nitrous Oxide.

OBJECTIVE: Combining intranasal fentanyl (IN FENT) with inhaled nitrous oxide (N2O) seems to have good properties for pediatric procedural sedation and analgesia (PSA). This study aims to assess the side effect rate of the combined use of IN FENT and N2O. METHODS: We performed a retrospective, single-center study. Patients treated in either the pediatric emergency department (PED) or the pediatric surgery outpatient clinic (PSOC) were included, if they received PSA with IN FENT and nitrous oxide with 50% oxygen (N2O 50%). RESULTS: Three hundred seventy-five patients were included over a period of 4 years. Median age was 9.4 years (range, 3.1 to 15.9) and 39% of patients were female. Overall side effect rate was 30% (114 patients). Most frequent was dizziness (n = 63, 17%; 95% CI, 13-21), followed by nausea (n = 23, 6%; 95% CI, 4-9) and emesis (n = 14, 4%; 95% CI, 2-6), with 35 patients having either nausea and/or emesis (9%; 95% CI, 7-13). No serious side effects were recorded (0%; 95% CI, 0-0.1). Of 298 patients with information regarding satisfaction, 280 patients would like the same sedation for a similar procedure in the future (94%; 95% CI, 90-96). We found no relation between previously described risk factors and emesis and/or nausea. CONCLUSIONS: N2O 50% combined with IN FENT can be recommended as an effective and safe treatment in the PED and the PSOC. While the side effect rate, primarily dizziness, nausea and emesis was substantial, antiemetic prophylaxis is not indicated owing to the overall low incidence of nausea and emesis.

Targeting SHP2 phosphatase in breast cancer overcomes RTK-mediated resistance to PI3K inhibitors.

BACKGROUND: PI3K signaling is frequently activated in breast cancer and is targeted by PI3K inhibitors. However, resistance of tumor cells to PI3K inhibition, often mediated by activated receptor tyrosine kinases, is commonly observed and reduces the potency of PI3K inhibitors. Therefore, new treatment strategies to overcome resistance to PI3K inhibitors are urgently needed to boost their efficacy. The phosphatase SHP2, which plays a crucial role in mediating signal transduction between receptor tyrosine kinases and both the PI3K and MAPK pathways, is a potential target for combination treatment. METHODS: We tested combinations of PI3K and SHP2 inhibitors in several experimental breast cancer models that are resistant to PI3K inhibition. Using cell culturing, biochemical and genetic approaches, we evaluated tumor cell proliferation and signaling output in cells treated with PI3K and SHP2 inhibitors. RESULTS: Combination treatment with PI3K and SHP2 inhibitors counteracted both acquired and intrinsic breast cancer cell resistance to PI3K inhibition that is mediated by activated receptor tyrosine kinases. Dual PI3K and SHP2 inhibition blocked proliferation and led to sustained inactivation of PI3K and MAPK signaling, where resistant cells rapidly re-activated these pathways upon PI3K inhibitor monotreatment. In addition, we demonstrate that overexpression of SHP2 induced resistance to PI3K inhibition, and that SHP2 was frequently activated during the development of PI3K inhibitor resistance after prolonged treatment of sensitive cells. CONCLUSIONS: Our results highlight the importance of SHP2 as a player in resistance to PI3K inhibitors. Combination treatment with PI3K and SHP2 inhibitors could pave the way for significant improvements in therapies for breast cancer.

YAP and ß-Catenin Cooperate to Drive Oncogenesis in Basal Breast Cancer.

Targeting cancer stem cells (CSC) can serve as an effective approach toward limiting resistance to therapies. While basal-like (triple-negative) breast cancers encompass cells with CSC features, rational therapies remain poorly established. We show here that the receptor tyrosine kinase Met promotes YAP activity in basal-like breast cancer and find enhanced YAP activity within the CSC population. Interfering with YAP activity delayed basal-like cancer formation, prevented luminal to basal transdifferentiation, and reduced CSC. YAP knockout mammary glands revealed a decrease in ß-catenin target genes, suggesting that YAP is required for nuclear ß-catenin activity. Mechanistically, nuclear YAP interacted with ß-catenin and TEAD4 at gene regulatory elements. Proteomic patient data revealed an upregulation of the YAP signature in basal-like breast cancers. Our findings demonstrate that in basal-like breast cancers, ß-catenin activity is dependent on YAP signaling and controls the CSC program. These findings suggest that targeting the YAP/TEAD4/ß-catenin complex offers a potential therapeutic strategy for eradicating CSCs in basal-like breast cancers. SIGNIFICANCE: These findings show that YAP cooperates with ß-catenin in basal-like breast cancer to regulate CSCs and that targeting this interaction may be a novel CSC therapy for patients with basal-like breast cancer. GRAPHICAL ABSTRACT: http://cancerres.aacrjournals.org/content/canres/81/8/2116/F1.large.jpg.

Role of tip apices in scanning force spectroscopy on alkali halides at room temperature-chemical nature of the tip apex and atomic-scale deformations.

We have revealed processes of the tip apex distortion in the measurements of non-contact scanning force microscopy. High-spatial-resolution two-dimensional force mapping on KCl(100) surfaces for a large number of tips, seven tips, enabled us to see the complex behavior of the tip apex distortion. The tips are from Si without additional coating, but are altered by the tip-sample interaction and show the behavior of different atomic species. On the KCl(001) surfaces, the tip apex, consisting of K and Cl atoms or of Si, distorted several times while changing the distance even in a weak attractive region. There are variations in rigidity of the tip apex, but all tips distorted in the small attractive region. This complex behavior was categorized in patterns by our analyses. We compare the experimental force-distance data to atomistic simulations using rigid KCl-terminated tips and KCl-terminated tips with an additional KCl-pair designed to perform atomic jumps. We also compare the experimental force-distance data to first principles simulations using Si tips. We mainly find K-terminated tips and Si-terminated tips. We find that Si tips show only one force minimum whereas KCl-terminated tips show two force minima in line with the stronger rigidity of Si compared to KCl. At room temperature, the tip apex atoms can perform atomic jumps that change the atomic configuration of the tip apex.

Electrostatic forces above graphene nanoribbons and edges interpreted as partly hydrogen-free.

Graphene nanoribbons' electronic transport properties strongly depend on the type of edge, armchair, zigzag or other, and on edge functionalization that can be used for band-gap engineering. For only partly hydrogenated edges interesting magnetic properties are predicted. Electric charge accumulates at edges and corners. Scanning force microscopy has so far shown the centre of graphene nanoribbons with atomic resolution using a quartz crystal tuning fork sensor of high stiffness. Weak long-range electrostatic forces related to the charge accumulation on the edges of graphene nanoribbons could not be imaged so far. Here, we show the electrostatic forces at the corners and edges of graphene nanoribbons are amenable to measurement. We use soft cantilevers and a bimodal imaging technique to combine enhanced sensitivity to weak long-range electrostatic forces with the high resolution of the second-frequency shift. Additionally, in our work the edges of the nanoribbons are mainly hydrogen-free, opening to the route to investigations of partly hydrogenated magnetic nanoribbons.

Near-equilibrium measurement of quantum size effects using Kelvin probe force microscopy.

In nano-structures such as thin films electron confinement results in the quantization of energy levels in the direction perpendicular to the film. The discretization of the energy levels leads to the oscillatory dependence of many properties on the film thickness due to quantum size effects. Pb on Si(111) is a specially interesting system because a particular relationship between the Pb atomic layer thickness and its Fermi wavelength leads to a periodicity of the oscillation of two atomic layers. Here, we demonstrate how the combination of scanning force microscopy (SFM) and Kelvin probe force microscopy (KPFM) provides a reliable method to monitor the quantum oscillations in the work function of Pb ultra-thin film nano-structures on Si(111). Unlike other techniques, with SFM/KPFM we directly address single Pb islands, determine their height while suppressing the influence of electrostatic forces, and, in addition, simultaneously evaluate their local work function by measurements close to equilibrium, without current-dependent and non-equilibrium effects. Our results evidence even-odd oscillations in the work function as a function of the film thickness that decay linearly with the film thickness, proving that this method provides direct and precise information on the quantum states.

Cancer Stem Cells Regulate Cancer-Associated Fibroblasts via Activation of Hedgehog Signaling in Mammary Gland Tumors.

Many tumors display intracellular heterogeneity with subsets of cancer stem cells (CSC) that sustain tumor growth, recurrence, and therapy resistance. Cancer-associated fibroblasts (CAF) have been shown to support and regulate CSC function. Here, we investigate the interactions between CSCs and CAFs in mammary gland tumors driven by combined activation of Wnt/ß-catenin and Hgf/Met signaling in mouse mammary epithelial cells. In this setting, CSCs secrete the Hedgehog ligand SHH, which regulate CAFs via paracrine activation of Hedgehog signaling. CAFs subsequently secrete factors that promote expansion and self-renewal of CSCs. In vivo treatment of tumors with the Hedgehog inhibitor vismodegib reduce CAF and CSC expansion, resulting in an overall delay of tumor formation. Our results identify a novel intracellular signaling module that synergistically regulates CAFs and CSCs. Targeting CAFs with Hedgehog inhibitors may offer a novel therapeutic strategy against breast cancer. Cancer Res; 77(8); 2134-47. ©2017 AACR.

Atomic-Scale Imaging of the Surface Dipole Distribution of Stepped Surfaces.

Stepped well-ordered semiconductor surfaces are important as nanotemplates for the fabrication of 1D nanostructures. Therefore, a detailed understanding of the underlying stepped substrates is crucial for advances in this field. Although measurements of step edges are challenging for scanning force microscopy (SFM), here we present simultaneous atomically resolved SFM and Kelvin probe force microscopy (KPFM) images of a silicon vicinal surface. We find that the local contact potential difference is large at the bottom of the steps and at the restatoms on the terraces, whereas it drops at the upper part of the steps and at the adatoms on the terraces. For the interpretation of the data we performed density functional theory (DFT) calculations of the surface dipole distribution. The DFT images accurately reproduce the experiments even without including the tip in the calculations. This underlines that the high-resolution KPFM images are closely related to intrinsic properties of the surface and not only to tip-surface interactions.

Shp2 signaling suppresses senescence in PyMT-induced mammary gland cancer in mice.

In this study, we have used techniques from cell biology, biochemistry, and genetics to investigate the role of the tyrosine phosphatase Shp2 in tumor cells of MMTV-PyMT mouse mammary glands. Genetic ablation or pharmacological inhibition of Shp2 induces senescence, as determined by the activation of senescence-associated ß-gal (SA-ß-gal), cyclin-dependent kinase inhibitor 1B (p27), p53, and histone 3 trimethylated lysine 9 (H3K9me3). Senescence induction leads to the inhibition of self-renewal of tumor cells and blockage of tumor formation and growth. A signaling cascade was identified that acts downstream of Shp2 to counter senescence: Src, focal adhesion kinase, and Map kinase inhibit senescence by activating the expression of S-phase kinase-associated protein 2 (Skp2), Aurora kinase A (Aurka), and the Notch ligand Delta-like 1 (Dll1), which block p27 and p53. Remarkably, the expression of Shp2 and of selected target genes predicts human breast cancer outcome. We conclude that therapies, which rely on senescence induction by inhibiting Shp2 or controlling its target gene products, may be useful in blocking breast cancer.

Domain wall pinning in ultra-narrow electromigrated break junctions.

The study of magnetic domain walls in constrained geometries is an important topic, yet when dealing with extreme nanoscale magnetic systems artefacts can often dominate the measurements and obscure the effects of intrinsic magnetic origin. In this work we study the evolution of domain wall depinning in electromigrated ferromagnetic junctions which are both initially fabricated and subsequently tailored in-situ in clean ultra-high vacuum conditions. Carefully designed Ni(80)Fe(20) (Permalloy) notched half-ring structures are fabricated and investigated as a function of constriction width by tailoring the size of the contact using controlled in-situ electromigration. It is found that the domain wall pinning strength is increased on reducing the contact size in line with a reduction of the wall energy in narrower constrictions. Furthermore, the angular dependency and symmetry of the depinning field is measured to determine the full pinning potential for a domain wall in a system with a narrow constriction.

Epitaxial growth of pentacene on alkali halide surfaces studied by Kelvin probe force microscopy.

In the field of molecular electronics, thin films of molecules adsorbed on insulating surfaces are used as the functional building blocks of electronic devices. Control of the structural and electronic properties of the thin films is required for reliably operating devices. Here, noncontact atomic force and Kelvin probe force microscopies have been used to investigate the growth and electrostatic landscape of pentacene on KBr(001) and KCl(001) surfaces. We have found that, together with molecular islands of upright standing pentacene, a new phase of tilted molecules appears near step edges on KBr. Local contact potential differences (LCPD) have been studied with both Kelvin experiments and density functional theory calculations. Our images reveal that differently oriented molecules display different LCPD and that their value is independent of the number of molecular layers. These results point to the formation of an interface dipole, which may be explained by a partial charge transfer from the pentacene to the surface. Moreover, the monitoring of the evolution of the pentacene islands shows that they are strongly affected by dewetting: Multilayers build up at the expense of monolayers, and in the Kelvin images, previously unknown line defects appear, which reveal the epitaxial growth of pentacene crystals.

Combined Wnt/ß-catenin, Met, and CXCL12/CXCR4 signals characterize basal breast cancer and predict disease outcome.

Prognosis for patients with estrogen-receptor (ER)-negative basal breast cancer is poor, and chemotherapy is currently the best therapeutic option. We have generated a compound-mutant mouse model combining the activation of ß-catenin and HGF (Wnt-Met signaling), which produced rapidly growing basal mammary gland tumors. We identified the chemokine system CXCL12/CXCR4 as a crucial driver of Wnt-Met tumors, given that compound-mutant mice also deficient in the CXCR4 gene were tumor resistant. Wnt-Met activation rapidly expanded a population of cancer-propagating cells, in which the two signaling systems control different functions, self-renewal and differentiation. Molecular therapy targeting Wnt, Met, and CXCR4 in mice significantly delayed tumor development. The expression of a Wnt-Met 322 gene signature was found to be predictive of poor survival of human patients with ER-negative breast cancers. Thus, targeting CXCR4 and its upstream activators, Wnt and Met, might provide an efficient strategy for breast cancer treatment.

Domain-wall induced large magnetoresistance effects at zero applied field in ballistic nanocontacts.

We determine magnetoresistance effects in stable and clean Permalloy nanocontacts of variable cross section, fabricated by UHV deposition and in situ electromigration. To ascertain the magnetoresistance (MR) effects originating from a magnetic domain wall, we measure the resistance values with and without such a wall at zero applied field. In the ballistic transport regime, the MR ratio reaches up to 50% and exhibits a previously unobserved sign change. Our results can be reproduced by recent atomistic calculations for different atomic configurations of the nanocontact, highlighting the importance of the detailed atomic arrangement for the MR effect.

Molecular-resolution imaging of pentacene on KCl(001).

The growth of pentacene on KCl(001) at submonolayer coverage was studied by dynamic scanning force microscopy. At coverages below one monolayer pentacene was found to arrange in islands with an upright configuration. The molecular arrangement was resolved in high-resolution images. In these images two different types of patterns were observed, which switch repeatedly. In addition, defects were found, such as a molecular vacancy and domain boundaries.

Distinct requirements for Gab1 in Met and EGF receptor signaling in vivo.

Gab1 is a multiadaptor protein that has been shown to be required for multiple processes in embryonic development and oncogenic transformation. Gab1 functions by amplifying signal transduction downstream of various receptor tyrosine kinases through recruitment of multiple signaling effectors, including phosphatidylinositol 3-kinase and Shp2. Until now, the functional significance of individual interactions in vivo was not known. Here we have generated knockin mice that carry point mutations in either the P13K or Shp2 binding sites of Gab1. We show that different effector interactions with Gab1 play distinct biological roles downstream of Gab1 during the development of different organs. Recruitment of phosphatidylinositol 3-kinase by Gab1 is essential for EGF receptor-mediated embryonic eyelid closure and keratinocyte migration, and the Gab1-Shp2 interaction is crucial for Met receptor-directed placental development and muscle progenitor cell migration to the limbs. Furthermore, we investigate the dual association of Gab1 with the Met receptor. By analyzing knockin mice with mutations in the Grb2 or Met binding site of Gab1, we show that the requirements for Gab1 recruitment to Met varies in different biological contexts. Either the direct or the indirect interaction of Gab1 with Met is sufficient for Met-dependent muscle precursor cell migration, whereas both modes of interaction are required and neither is sufficient for placenta development, liver growth, and palatal shelf closure. These data demonstrate that Gab1 induces different biological responses through the recruitment of distinct effectors and that different modes of recruitment for Gab1 are required in different organs.

[Treatment effects of parent management training for ADHD]. / Die Wirksamkeit eines Gruppentrainings für Eltern hyperkinetischer Kinder.

OBJECTIVES: The study investigated the efficacy of a programme of parent management training in groups for the parents of ADHD children that is based on a cognitive behavioural concept. METHODS: The parents of 16 ADHD children participated in group training over a ten-week period. Children were medicated with methylphenidate and received consultations. 17 other ADHD children (control group) benefited only from medication with methylphenidate plus consultation; however, their parents did not participate in any group training. In both groups, assessment was done by means of parent questionnaires: in the experimental group to assess the core symptoms of ADHD, homework problems and global problems that occur in families with ADHD; in the control group to assess only the core symptoms of ADHD. RESULTS: The training significantly reduced core symptoms of ADHD, homework problems and global problematic situations in the experimental group. In comparison to the control group, the core symptoms of ADHD decreased more strongly in the experimental group. This effect failed to reach the significance level of p < 0.05 (Mann-Whitney U-test). However, the verification with the linear model showed a significant difference between the two groups on the factor "hyperactivity index". CONCLUSIONS: Parent management training in groups was a useful adjunct in the treatment of ADHD children.