Regulation of protumorigenic pathways by Insulin like growth factor binding protein2 and its association along with beta-catenin in breast cancer lymph node metastasis

Background: Insulin like growth factor binding proteins modulate the mitogenic and pro survival effects of IGF. Elevated expression of IGFBP2 is associated with progression of tumors that include prostate, ovarian, glioma among others. Though implicated in the progression of breast cancer, the molecular mechanisms involved in IGFBP2 actions are not well defined. This study investigates the molecular targets and biological pathways targeted by IGFBP2 in breast cancer. Methods: Transcriptome analysis of breast tumor cells (BT474) with stable knockdown of IGFBP2 and breast tumors having differential expression of IGFBP2 by immunohistochemistry was performed using microarray. Differential gene expression was established using R-Bioconductor package. For validation, gene expression was determined by qPCR. Inhibitors of IGF1R and integrin pathway were utilized to study the mechanism of regulation of beta-catenin. Immunohistochemical and immunocytochemical staining was performed on breast tumors and experimental cells, respectively for beta-catenin and IGFBP2 expression. Results: Knockdown of IGFBP2 resulted in differential expression of 2067 up regulated and 2002 down regulated genes in breast cancer cells. Down regulated genes principally belong to cell cycle, DNA replication, repair, p53 signaling, oxidative phosphorylation, Wnt signaling. Whole genome expression analysis of breast tumors with or without IGFBP2 expression indicated changes in genes belonging to Focal adhesion, Map kinase and Wnt signaling pathways. Interestingly, IGFBP2 knockdown clones showed reduced expression of beta-catenin compared to control cells which was restored upon IGFBP2 re-expression. The regulation of beta-catenin by IGFBP2 was found to be IGF1R and integrin pathway dependent. Furthermore, IGFBP2 and beta-catenin are co-ordinately overexpressed in breast tumors and correlate with lymph node metastasis. Conclusion: This study highlights regulation of beta-catenin by IGFBP2 in breast cancer cells and most importantly, combined expression of IGFBP2 and beta-catenin is associated with lymph node metastasis of breast tumors.

Probing p300/CBP associated Factor (PCAF)-dependent pathways with a small molecule inhibitor

PCAF (KAT2B) belongs to the GNAT family of lysine acetyltransferases (KAT) and specifically acetylates the histone H3K9 residue and several nonhistone proteins. PCAF is also a transcriptional coactivator. Due to the lack of a PCAF KAT-specific small molecule inhibitor, the exclusive role of the acetyltransferase activity of PCAF is not well understood. Here, we report that a natural compound of the hydroxybenzoquinone class, embelin, specifically inhibits H3Lys9 acetylation in mice and inhibits recombinant PCAF-mediated acetylation with near complete specificity in vitro. Furthermore, using embelin, we have identified the gene networks that are regulated by PCAF during muscle differentiation, further highlighting the broader regulatory functions of PCAF in muscle differentiation in addition to the regulation via MyoD acetylation.

Molecular flexibility tuned emission in ``V'' shaped naphthalimides: Hg(II) detection and aggregation-induced emission enhancement (AIEE)

Four ``V'' shaped 1,8-naphthalimides (1-4) have been synthesized and their fluorescence quantum-yields correlated to their molecular flexibility. The correlation was used for detection of Hg(II) via a chemodosimetric approach. 4 was found to be an AIE active molecule with the formation of fluorescent nanoaggregates.

On the toughness enhancement in hydroxyapatite-based composites

Among various biologically compatible materials, hydroxyapatite (HA) has excellent bioactivity/osteointegration properties and therefore has been extensively investigated for biomedical applications. However, its inferior fracture toughness limits the wider applications of monolithic HA as a load-bearing implant. To this end, HA-based biocomposites have been developed to improve their mechanical properties (toughness and strength) without compromising biocompatibility. Despite significant efforts over last few decades, the toughness of HA-based composites could not be enhanced beyond 1.5-2 MPa m(1/2), even when measured using indentation techniques. In this perspective, the present work demonstrates how spark plasma sintering can be effectively utilized to develop hydroxyapatite titanium (HA-Ti) composites with varying amounts of Ti (5, 10 and 20 wt.%) with extremely high single edge V-notch beam fracture toughness (4-5 MPa m(1/2)) along with a good combination of elastic modulus and flexural strength. Despite predominant retention of HA and Ti, the combination of critical analysis of X-ray diffraction and transmission electron microscopy investigation confirmed the formation of the CaTi4(PO4)(6) phase with nanoscale morphology at the HA/Ti interface and the formation of such a phase has been discussed in reference to possible sintering reactions. The variations in the measured fracture toughness and work of fracture with Ti addition to the HA matrix were further rationalized using the analytical models of crack bridging as well as on the basis of the additional contribution from crack deflection. The present work opens up the opportunity to further enhance the toughness beyond 5 MPa m(1/2) by microstructural designing with the desired combination of toughening phases. (C) 2013 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Impact of substrate nitridation on the photoluminescence and photovoltaic characteristics of GaN grown on p-Si (100) by molecular beam epitaxy

This report focuses on the structural and optical properties of the GaN films grown on p-Si (100) substrates along with photovoltaic characteristics of GaN/p-Si heterojunctions fabricated with substrate nitridation and in absence of substrate nitridation. The high resolution X-ray diffraction (HRXRD), atomic force microscopy (AFM), Raman and photoluminescence (PL) spectroscopic studies reveal that the significant enhancement in the structural as well as in the optical properties of GaN epifilms grown with silicon nitride buffer layer when compared with the sample grown without silicon nitride buffer layer. The low temperature PL shows a free excitonic (FX) emission peak at 3.51 eV at the temperature of 5 K with a very narrow line width of 35 meV. Temperature dependent PL spectra follow the Varshni equation well and peak energy blue shifts by similar to 63 meV from 300 to 5 K. Raman data confirms the strain free nature and reasonably good crystallinity of the films. The GaN/p-Si heterojunctions fabricated without substrate nitridation show a superior photovoltaic performance compared to the devices fabricated in presence of substrate nitridation. The discussions have been carried out on the junction properties. Such single junction devices exhibit a promising fill factor and conversion efficiency of 23.36 and 0.12 %, respectively, under concentrated AM1.5 illumination.

Effect of a natural mutation in the 5 ` untranslated region on the translational control of p53 mRNA

Tumor-suppressor protein p53, the `guardian of the genome', is critical in maintaining cellular homeostasis and genomic stability. Earlier, we have reported the discovery of internal ribosome entry sites (IRESs) within the p53 mRNA that regulate the translation of the full length and its N-terminal-truncated isoform, Delta N-p53. Polypyrimidine tract-binding protein (PTB) is an IRES trans-acting factor that positively regulates the IRES activities of both p53 isoforms by relocating from nucleus to the cytoplasm during stress conditions. Here we have demonstrated the putative contact points of PTB on the p53 IRES RNA. Studies on mutations that occur naturally in the 5' untranslated region (5' UTR) in p53 mRNA were lacking. We have investigated a naturally occurring C-to-T single-nucleotide polymorphism (SNP) first reported in human melanoma tumors. This SNP is at position 119 in the 5' UTR of p53 mRNA and we demonstrate that it has consequences on the translational control of p53. Introduction of this SNP has led to decrease in cap-independent translation from p53 5' UTR in bicistronic reporter assay. Further, the effects of this SNP on cap-independent translation have been studied in the context of p53 cDNA as well. Interestingly, the 5' UTR with this SNP has shown reduced binding to PTB that can be corroborated to its weaker IRES activity. Previously, it has been shown that G2-M checkpoint, DNA-damaging stress and oncogenic insult favor IRES-mediated translation. Under similar conditions, we demonstrate that this SNP interferes with the enhancement of the IRES activity of the 5' UTR. Taken together, the results demonstrate for the first time that SNP in the 5' UTR of the p53 mRNA might have a role in translational control of this critical tumor-suppressor gene.

Tuning the ferromagnetic transition temperature in La0.5Sr0.5CoO3 thin films

Epitaxial La0.5Sr0.5CoO3 (LSCO) thin films are grown on LaAlO3 (100) and SrTiO3 (100) substrates by pulsed laser ablation. By tuning the growth parameters, we are able to enhance the ferromagnetic transition temperature (T-C) up to 262 K as evident from ac susceptibility, dc magnetization, and resistivity measurements. The magnitude of T-C is the same as that for the bulk stoichiometric LSCO illustrating the high quality of the grown films. Detailed structural analysis clearly reveals that the induced strain in the LSCO film has no role behind this enhancement; in fact, the determining factor is the oxygen stoichiometry. The films also exhibit ageing effect as the T-C decreases with time. This is considered in terms of gradual change in the oxygen stoichiometry through diffusion process as the time progresses. (C) 2013 AIP Publishing LLC.

Role of silica nanoparticles in conductivity enhancement of nanocomposite solid polymer electrolytes: (PEG(x) NaBr): ySiO(2)

Nanocomposite solid polymer electrolytes (NCSPEs) with conducting species other than Li ions are being investigated for solid-state battery applications. Pristine solid polymer electrolytes (SPEs) do not show ionic conductivity suitable for batteries. Addition of inert fillers to SPEs is known to enhance the ionic conductivity. In this paper, we present the role of silica nanoparticles in enhancing the ionic conductivity in NCSPEs with sodium as conducting species. Sodium bromide is complexed with the host polyethylene glycol polymer by solution cast method and silica nanoparticles (SiO2, average particle size 7 nm) are incorporated into the complex in small amounts. The composites are characterized by powder XRD and IR spectroscopy. Conductivity measurements are undertaken as a function of concentration of salt and also as a function of temperature using impedance spectroscopy. Addition of silica nanoparticles shows an enhancement in conductivity by 1-2 orders of magnitude. The results are discussed in terms of interaction of nanoparticles with the nonconducting anions.

Parametrisation-free determination of the shape parameters for the pion electromagnetic form factor

Recent data from high-statistics experiments that have measured the modulus of the pion electromagnetic form factor from threshold to relatively high energies are used as input in a suitable mathematical framework of analytic continuation to find stringent constraints on the shape parameters of the form factor at t = 0. The method uses also as input a precise description of the phase of the form factor in the elastic region based on Fermi-Watson theorem and the analysis of the pi pi scattering amplitude with dispersive Roy equations, and some information on the spacelike region coming from recent high precision experiments. Our analysis confirms the inconsistencies of several data on the modulus, especially from low energies, with analyticity and the input phase, noted in our earlier work. Using the data on the modulus from energies above 0.65 GeV, we obtain, with no specific parametrisation, the prediction < r(pi)(2)> is an element of (0.42, 0.44) fm(2) for the charge radius. The same formalism leads also to very narrow allowed ranges for the higher-order shape parameters at t = 0, with a strong correlation among them.

Base Triggered Enhancement of First Hyperpolarizability of a Keto-Enol Tautomer

This work describes the base triggered enhancement of first hyperpolarizability of a tautomeric organic molecule, namely, benzoylacetanilide (BA). We have used the hyper-Rayleigh scattering technique to measure the first hyperpolarizability (beta) of BA which exists in the pure keto form in water and as a keto-enol tautomer in ethanol. Its anion exists in equilibrium with the keto and enol forms at pH 11 in aqueous solution. The beta value of the anion form is 709 X 10(-30) esu, whereas that of the enol is 232 x 10(-3) esu and of the keto is 88 X 10(-30) esu. There is an enhancement of beta by similar to 8 times for the anion and similar to 3 times for the enol compared to the keto form. All these are achieved by altering the equilibrium between the three forms of BA by simple means. MP2 calculations reproduce the experimental trend, but the computed beta values are much lower than the measured values. DFT calculations with the standard B3LYP functional could not predict the right order in the beta values. The difference between experimental and calculated values is, perhaps, due to the fact that electron correlation effects are important in computing optical nonlinearities of large organic molecules and MP2 and B3LYP calculations done here for different forms of BA could not account for such effects adequately.

Molecular flexibility of Mycobacterium tuberculosis ribosome recycling factor and its functional consequences: An exploration involving mutants

Internal mobility of the two domain molecule of ribosome recycling factor (RRF) is known to be important for its action. Mycobacterium tuberculosis RRF does not complement E. coli for its deficiency of RRF (in the presence of E. coli EF-G alone). Crystal structure had revealed higher rigidity of the M. tuberculosis RRF due to the presence of additional salt bridges between domains. Two inter-domain salt bridges and one between the linker region and the domain containing C-terminal residues were disrupted by appropriate mutations. Except for a C-terminal deletion mutant, all mutants showed RRF activity in E. coli when M. tuberculosis EF-G was also co-expressed. The crystal structures of the point mutants, that of the C-terminal deletion mutant and that of the protein grown in the presence of a detergent, were determined. The increased mobility resulting from the disruption of the salt bridge involving the hinge region allows the appropriate mutant to weakly complement E. coli for its deficiency of RRF even in the absence of simultaneous expression of the mycobacterial EF-G. The loss of activity of the C-terminal deletion mutant appears to be partly due to the rigidification of the molecule consequent to changes in the hinge region.

Bounds on the spacelike pion electromagnetic form factor from analyticity and unitarity

We use the recently measured accurate BaBaR data on the modulus of the pion electromagnetic form factor,Fπ(t), up to an energy of 3 GeV, the I=1P-wave phase of the π π scattering ampli-tude up to the ω−π threshold, the pion charge radius known from Chiral Perturbation Theory,and the recently measured JLAB value of Fπ in the spacelike region at t=−2.45GeV2 as inputs in a formalism that leads to bounds on Fπ in the intermediate spacelike region. We compare our constraints with experimental data and with perturbative QCD along with the results of several theoretical models for the non-perturbative contribution s proposed in the literature.

Tuning the parameters to establish Quenching and enhancement regimes in hybrid Gold nanoparticles and CdSe Quantum dots monolayer

The multi-component nanomaterials combine the individual properties and give rise to emergent phenomenon. Optical excitations in such hybrid nonmaterial's ( for example Exciton in semiconductor quantum dots and Plasmon in Metal nanomaterials) undergo strong weak electromagnetic coupling. Such exciton-plasmon interactions allow design of absorption and emission properties, control of nanoscale energy-transfer processes, and creation of new excitations in the strong coupling regime.This Exciton plasmon interaction in hybrid nanomaterial can lead to both enhancement in the emission as well as quenching. In this work we prepared close-packed hybrid monolayer of thiol capped CdSe and gold nanoparticles. They exhibit both the Quenching and enhancements the in PL emission.The systematic variance of PL from such hybrid nanomaterials monolayer is studied by tuning the Number ratio of Gold per Quantum dots, the surface density of QDs and the spectral overlap of emission spectrum of QD and absorption spectrum of Gold nanoparticles. Role of Localized surface Plasmon which not only leads to quenching but strong enhancements as well, is explored.

Bactericidal effect of silver-reinforced carbon nanotube and hydroxyapatite composites

Bacterial infection remains an important risk factor after orthopedic surgery. The present paper reports the synthesis of hydroxyapatite-silver (HA-Ag) and carbon nanotube-silver (CNT-Ag) composites via spark plasma sintering (SPS) route. The retention of the initial phases after SPS was confirmed by phase analysis using X-ray diffraction and Raman spectroscopy. Energy dispersive spectrum analysis showed that Ag was distributed uniformly in the CNT/HA matrix. The breakage of CNTs into spheroid particles at higher temperatures (1700 degrees C) is attributed to the Rayleigh instability criterion. Mechanical properties (hardness and elastic modulus) of the samples were evaluated using nanoindentation testing. Ag reinforcement resulted in the enhancement of hardness (by similar to 15%) and elastic modulus (similar to 5%) of HA samples, whereas Ag reinforcement in CNT, Ag addition does not have much effect on hardness (0.3 GPa) and elastic modulus (5 GPa). The antibacterial tests performed using Escherichia coli and Staphylococcus epidermidis showed significant decrease (by similar to 65-86%) in the number of adhered bacteria in HA/CNT composites reinforced with 5% Ag nanoparticles. Thus, Ag-reinforced HA/CNT can serve as potential antibacterial biocomposites.

Insulin like growth factor binding protein 4 promotes GBM progression and regulates key factors involved in EMT and invasion

Insulin like growth factor binding protein 4 (IGFBP4) regulates growth and development of tissues and organs by negatively regulating IGF signaling. Among most cancers, IGFBP4 has growth inhibitory role and reported as a down-regulated gene, except for renal cell carcinoma, wherein IGFBP4 promotes tumor progression. IGFBP4 expression has been shown to be higher in increasing grades of astrocytoma. However, the functional role of IGFBP4 in gliomas has not been explored. Surgical biopsies of 20 normal brain and 198 astrocytoma samples were analyzed for IGFBP4 expression by qRT-PCR. Highest expression of IGFBP4 mRNA was seen in GBM tumors compared to control brain tissues (median log2 of 2.035, p < 0.0001). Immunohistochemical analysis of 53 tissue samples revealed predominant nuclear staining of IGFBP4, seen maximally in GBMs when compared to DA and AA tumors (median LI = 29.12 +/- A 16.943, p < 0.001). Over expression of IGFBP4 in U343 glioma cells resulted in up-regulation of molecules involved in tumor growth, EMT and invasion such as pAkt, pErk, Vimentin, and N-cadherin and down-regulation of E-cadherin. Functionally, IGFBP4 over expression in these cells resulted in increased proliferation, migration and invasion as assessed by MTT, transwell migration, and Matrigel invasion assays. These findings were confirmed upon IGFBP4 knockdown in U251 glioma cells. Our data suggest a pro-tumorigenic role for IGFBP4 in glioma.