Novel functional interaction between the plasma membrane Ca2+ pump 4b and the proapoptotic tumor suppressor Ras-associated factor 1 (RASSF1)

Plasma membrane calmodulin-dependent calcium ATPases (PMCAs) are enzymatic systems implicated in the extrusion of calcium from the cell. We and others have previously identified molecular interactions between the cytoplasmic COOH-terminal end of PMCA and PDZ domain-containing proteins. These interactions suggested a new role for PMCA as a modulator of signal transduction pathways. The existence of other intracellular regions in the PMCA molecule prompted us to investigate the possible participation of other domains in interactions with different partner proteins. A two-hybrid screen of a human fetal heart cDNA library, using the region 652-840 of human PMCA4b (located in the catalytic, second intracellular loop) as bait, revealed a novel interaction between PMCA4b and the tumor suppressor RASSF1, a Ras effector protein involved in H-Ras-mediated apoptosis. Immunofluorescence co-localization, immunoprecipitation, and glutathione S-transferase pull-down experiments performed in mammalian cells provided further confirmation of the physical interaction between the two proteins. The interaction domain has been narrowed down to region 74-123 of RASSF1C (144-193 in RASSF1A) and 652-748 of human PMCA4b. The functionality of this interaction was demonstrated by the inhibition of the epidermal growth factor-dependent activation of the Erk pathway when PMCA4b and RASSF1 were co-expressed. This inhibition was abolished by blocking PMCA/RASSSF1 association with an excess of a green fluorescent protein fusion protein containing the region 50-123 of RASSF1C. This work describes a novel protein-protein interaction involving a domain of PMCA other than the COOH terminus. It suggests a function for PMCA4b as an organizer of macromolecular protein complexes, where PMCA4b could recruit diverse proteins through interaction with different domains. Furthermore, the functional association with RASSF1 indicates a role for PMCA4b in the modulation of Ras-mediated signaling.

IGF-Binding Protein 2 - Oncogene or Tumor Suppressor?

The role of insulin-like growth factor binding protein 2 (IGFBP2) in cancer is unclear. In general, IGFBP2 is considered to be oncogenic and its expression is often observed to be elevated in cancer. However, there are a number of conflicting reports in vitro and in vivo where IGFBP2 acts in a tumor suppressor manner. In this mini-review, we discuss the factors influencing the variation in IGFBP2 expression in cancer and our interpretation of these findings.

High-order-harmonic generation in benzene with linearly and circularly polarised laser pulses

High-order-harmonic generation in benzene is studied using a mixed quantum-classical approach in which the electrons are described using time-dependent density functional theory while the ions move classically. The interaction with both linearly and circularly polarised infra-red ($\lambda = 800$ nm) laser pulses of duration 10 cycles (26.7 fs) is considered. The effect of allowing the ions to move is investigated as is the effect of including self-interaction corrections to the exchange-correlation functional. Our results for circularly polarised pulses are compared with previous calculations in which the ions were kept fixed and self-interaction corrections were not included while our results for linearly polarised pulses are compared with both previous calculations and experiment. We find that even for the short duration pulses considered here, the ionic motion greatly influences the harmonic spectra. While ionization and ionic displacements are greatest when linearly polarised pulses are used, the response to circularly polarised pulses is almost comparable, in agreement with previous experimental results.

Comprehensive numerical modelling of the performance of a second harmonic generation stage coupled with a low-gain optical parametric amplifier

We present a comprehensive model for predicting the full performance of a second harmonic generation-optical parametric amplification system that aims at enhancing the temporal contrast of laser pulses. The model simultaneously takes into account all the main parameters at play in the system such as the group velocity mismatch, the beam divergence, the spectral content, the pump depletion, and the length of the nonlinear crystals. We monitor the influence of the initial parameters of the input pulse and the interdependence of the two related non-linear processes on the performance of the system and show its optimum configuration. The influence of the initial beam divergence on the spectral and the temporal characteristics of the generated pulse is discussed. In addition, we show that using a crystal slightly longer than the optimum length and introducing small delay between the seed and the pump ensures maximum efficiency and compensates for the spectral shift in the optical parametric amplification stage in case of chirped input pulse. As an example, calculations for bandwidth transform limited and chirped pulses of sub-picosecond duration in beta barium borate crystal are presented.

Comparative study of power converter topologies and control strategies for the harmonic performance of variable-speed wind turbine generator systems

Power converters play a vital role in the integration of wind power into the electrical grid. Variable-speed wind turbine generator systems have a considerable interest of application for grid connection at constant frequency. In this paper, comprehensive simulation studies are carried out with three power converter topologies: matrix, two-level and multilevel. A fractional-order control strategy is studied for the variable-speed operation of wind turbine generator systems. The studies are in order to compare power converter topologies and control strategies. The studies reveal that the multilevel converter and the proposed fractional-order control strategy enable an improvement in the power quality, in comparison with the other power converters using a classical integer-order control strategy. (C) 2010 Elsevier Ltd. All rights reserved.

Metal ions modulate the folding and stability of the tumor suppressor protein S100A2

Febs Journal (2009)276:1776-1786

Notch tumor suppressor function

Cancer development results from deregulated control of stem cell populations and alterations in their surrounding environment. Notch signaling is an important form of direct cell-cell communication involved in cell fate determination, stem cell potential and lineage commitment. The biological function of this pathway is critically context dependent. Here we review the pro-differentiation role and tumor suppressing function of this pathway, as revealed by loss-of-function in keratinocytes and skin, downstream of p53 and in cross-connection with other determinants of stem cell potential and/or tumor formation, such as p63 and Rho/CDC42 effectors. The possibility that Notch signaling elicits a duality of signals, involved in growth/differentiation control and cell survival will be discussed, in the context of novel approaches for cancer therapy

Inhibition of cell migration and invasion mediated by the TAT-RasGAP317-326 peptide requires the DLC1 tumor suppressor.

TAT-RasGAP317-326, a peptide corresponding to the 317-326 sequence of p120 RasGAP coupled with a cell-permeable TAT-derived peptide, sensitizes the death response of various tumor cells to several anticancer treatments. We now report that this peptide is also able to increase cell adherence, prevent cell migration and inhibit matrix invasion. This is accompanied by a marked modification of the actin cytoskeleton and focal adhesion redistribution. Interestingly, integrins and the small Rho GTP-binding protein, which are well-characterized proteins modulating actin fibers, adhesion and migration, do not appear to be required for the pro-adhesive properties of TAT-RasGAP317-326. In contrast, deleted in liver cancer-1, a tumor suppressor protein, the expression of which is often deregulated in cancer cells, was found to be required for TAT-RasGAP317-326 to promote cell adherence and inhibit migration. These results show that TAT-RasGAP317-326, besides its ability to favor tumor cell death, hampers cell migration and invasion.

PRDM1 is a tumor suppressor gene in natural killer cell malignancies.

Natural killer cell lymphoma (NKCL) constitutes a rare and aggressive form of non-Hodgkin lymphoma, and there is little insight into its pathogenesis. Here we show that PRDM1 is a tumor suppressor gene in NKCLs that is inactivated by a combination of monoallelic deletion and promoter CpG island hypermethylation. We observed monoallelic deletion of PRDM1 loci in 8 of 18 (44%) NKCL cases. The other allele showed significant promoter methylation in 12 of 17 (71%) cases. In support of its role as a tumor suppressor gene, the reconstitution of PRDM1 in PRDM1-null NK cell lines led to G2/M cell cycle arrest, increased apoptosis, and a strong negative selection pressure with progressive elimination of PRDM1-expressing cells, which was enhanced when IL-2 concentration is limiting. We observed a progressive increase in PRDM1 expression-in particular, PRDM1α-in normal NK cells in response to IL-2 and in normal NK cells activated with an engineered NK cell target, K562-Cl9-mb21, suggesting its role in NK cell homeostasis. In support of this role, knockdown of PRDM1 by shRNA in normal NK cells resulted in the positive selection of these cells. We identified MYC and 4-1BBL as targets of PRDM1 in NK cells. Disruption of homeostatic control by PRDM1 may be an important pathogenetic mechanism for NKCL.

On the formulation and the calculation of the harmonic contributions to the Debye-Waller factor in metals (sodium)

The algebraic expressions for the anharmonic contributions to the Debye-Waller factor up to 0(A ) and 0 L% ) £ where ^ is the scattering wave-vector] have been derived in a form suitable for cubic metals with small ion cores where the interatomic potential extends to many neighbours. This has been achieved in terms of various wave-vector dependent tensors, following the work of Shukla and Taylor (1974) on the cubic anharmonic Helmholtz free energy. The contribution to the various wave-vector dependent tensors from the coulomb and the electron-ion terms in the interatomic metallic potential has been obtained by the Ewald procedure. All the restricted multiple whole B r i l l o u i n zone (B.Z.) sums are reduced to single whole B.Z. sums by using the plane wave representation of the delta function. These single whole B.Z. sums are further reduced to the •%?? portion of the B.Z. following Shukla and Wilk (1974) and Shukla and Taylor (1974). Numerical calculations have been performed for sodium where the Born-Mayer term in the interatomic potential has been neglected because i t is small £ Vosko (1964)3 • *n o^er to compare our calculated results with the experimental results of Dawton (1937), we have also calculated the r a t io of the intensities at different temperatures for the lowest five reflections (110), (200), (220), (310) and (400) . Our calculated quasi-harmonic results agree reasonably well with the experimental results at temperatures (T) of the order of the Debye temperature ( 0 ). For T » © ^ 9 our calculated anharmonic results are found to be in good agreement with the experimental results.The anomalous terms in the Debye-Waller factor are found not to be negligible for certain reflections even for T ^ ©^ . At temperature T yy Op 9 where the temperature is of the order of the melting temperature (Xm) » "the anomalous terms are found to be important almost for all the f i ve reflections.

Regulation of BAP1 tumor suppressor complex by post-translational modifications

Le régulateur transcriptionnel BAP1 est une déubiquitinase nucléaire (DUB) dont le substrat est l’histone H2A modifiée par monoubiquitination au niveau des residus lysines 118 et 119 (K118/K119). Depuis les dernières années, BAP1 emerge comme un gene suppresseur de tumeur majeur. En effet, BAP1 est inactivé dans un plethore de maladies humaines héréditaires et sporadiques. Cependant, malgré l’accumulation significative des connaissances concernant l’occurrence, la pénétrance et l’impact des défauts de BAP1 sur le développement de cancers, ses mécanismes d’action et de régulation restent très peu compris. Cette étude est dédiée à la caractérisation moléculaire et fonctionnelle du complexe multi-protéique de BAP1 et se présente parmi les premiers travaux décrivant sa régulation par des modifications post-traductionnelles. D’abord, nous avons défini la composition du corps du complexe BAP1 ainsi que ses principaux partenaires d’interaction. Ensuite, nous nous sommes spécifiquement intéressés a investiguer d’avantage deux principaux aspects de la régulation de BAP1. Nous avons d’abord décrit l’inter-régulation entre deux composantes majeures du complexe BAP1, soit HCF-1 et OGT. D’une manière très intéressante, nous avons trouvé que le cofacteur HCF-1 est un important régulateur des niveaux protéiques d’OGT. En retour, OGT est requise pour la maturation protéolytique de HCF-1 en promouvant sa protéolyse par O-GlcNAcylation, un processus de régulation très important pour le bon fonctionnement de HCF-1. D’autre part, nous avons découvert un mécanisme unique de régulation de BAP1 médiée par l’ubiquitine ligase atypique UBE2O. en effet, UBE2O se caractérise par le fait qu’il s’agit aussi bien d’une ubiquitine conjuratrice et d’une ubiquitine ligase. UBE2O, multi-monoubiquitine BAP1 au niveau de son domaine NLS et promeut son exclusion du noyau, le séquestrant ainsi dans le cytoplasme. De façon importante, nos travaux ont permis de mettre de l’emphase sur le rôle de l’activité auto-catalytique de chacune de ces enzymes, soit l’activité d’auto-déubiquitination de BAP1 qui est requise pour la maintenance de sa localisation nucléaire ainsi que l’activité d’auto-ubiquitination d’UBE2O impliquée dans son transport nucléo-cytoplasmique. De manière significative, nous avons trouvé que des défauts au niveau de l’auto-déubiquitination de BAP1 due à des mutations associées à certains cancers indiquent l’importance d’une propre regulation de cette déubiquitinase pour les processus associés à la suppression de tumeurs.

Studies on some nonlinear optical phenomena-optical phase conjugation and continuous wave second harmonic generation

Nonlinear optics has emerged as a new area of physics , following the development of various types of lasers. A number of advancements , both theoretical and experimental . have been made in the past two decades . by scientists al1 over the world. However , onl y few scientists have attempted to study the experimental aspects of nonlinear optical phenomena i n I ndian laboratories. This thesis is the report of an attempt made in this direction. The thesis contains the details of the several investigations which the author has carried out in the past few years, on optical phase conjugation (OPC) and continuous wave CCVD second harmonic generation CSHG). OPC is a new branch of nonlinear optics, developed only in the past decade. The author has done a few experiments on low power OPC in dye molecules held in solid matrices, by making use of a degenerate four wave mixing CDFWND scheme. These samples have been characterised by studies on their absorption-spectra. fluorescence spectra. triplet lifetimes and saturation intensities. Phase conjugation efficiencies with r espect to the various parameters have been i nvesti gated . DFWM scheme was also employed i n achievi ng phase conjugation of a br oadband laser C Nd: G1ass 3 using a dye solution as the nonlinear medium.

Compact CPW-Fed Slot Antenna with Harmonic Suppression

A compact coplanar waveguide (CPW)-fed uniplanar antenna with harmonic suppression characteristics is presented. The above characteristics are achieved by properly modifying the ground plane and adjusting the signal strip of an open-ended CPW-fed transmission line. The simulated and experimental characteristics of the antenna are presented, compared, and discussed.

FEM based Virtual Prototyping and Design of Third Harmonic Excitation System for Low Voltage Brushless Alternators

Salient pole brushless alternators coupled to IC engines are extensively used as stand-by power supply units for meeting in- dustrial power demands. Design of such generators demands high power to weight ratio, high e ciency and low cost per KVA out- put. Moreover, the performance characteristics of such machines like voltage regulation and short circuit ratio (SCR) are critical when these machines are put into parallel operation and alterna- tors for critical applications like defence and aerospace demand very low harmonic content in the output voltage. While designing such alternators, accurate prediction of machine characteristics, including total harmonic distortion (THD) is essential to mini- mize development cost and time. Total harmonic distortion in the output voltage of alternators should be as low as possible especially when powering very sophis- ticated and critical applications. The output voltage waveform of a practical AC generator is replica of the space distribution of the ux density in the air gap and several factors such as shape of the rotor pole face, core saturation, slotting and style of coil disposition make the realization of a sinusoidal air gap ux wave impossible. These ux harmonics introduce undesirable e ects on the alternator performance like high neutral current due to triplen harmonics, voltage distortion, noise, vibration, excessive heating and also extra losses resulting in poor e ciency, which in turn necessitate de-rating of the machine especially when connected to non-linear loads. As an important control unit of brushless alternator, the excitation system and its dynamic performance has a direct impact on alternator's stability and reliability. The thesis explores design and implementation of an excitation i system utilizing third harmonic ux in the air gap of brushless al- ternators, using an additional auxiliary winding, wound for 1=3rd pole pitch, embedded into the stator slots and electrically iso- lated from the main winding. In the third harmonic excitation system, the combined e ect of two auxiliary windings, one with 2=3rd pitch and another third harmonic winding with 1=3rd pitch, are used to ensure good voltage regulation without an electronic automatic voltage regulator (AVR) and also reduces the total harmonic content in the output voltage, cost e ectively. The design of the third harmonic winding by analytic methods demands accurate calculation of third harmonic ux density in the air gap of the machine. However, precise estimation of the amplitude of third harmonic ux in the air gap of a machine by conventional design procedures is di cult due to complex geome- try of the machine and non-linear characteristics of the magnetic materials. As such, prediction of the eld parameters by conven- tional design methods is unreliable and hence virtual prototyping of the machine is done to enable accurate design of the third har- monic excitation system. In the design and development cycle of electrical machines, it is recognized that the use of analytical and experimental methods followed by expensive and in exible prototyping is time consum- ing and no longer cost e ective. Due to advancements in com- putational capabilities over recent years, nite element method (FEM) based virtual prototyping has become an attractive al- ternative to well established semi-analytical and empirical design methods as well as to the still popular trial and error approach followed by the costly and time consuming prototyping. Hence, by virtually prototyping the alternator using FEM, the important performance characteristics of the machine are predicted. Design of third harmonic excitation system is done with the help of results obtained from virtual prototype of the machine. Third harmonic excitation (THE) system is implemented in a 45 KVA ii experimental machine and experiments are conducted to validate the simulation results. Simulation and experimental results show that by utilizing third harmonic ux in the air gap of the ma- chine for excitation purposes during loaded conditions, triplen harmonic content in the output phase voltage is signi cantly re- duced. The prototype machine with third harmonic excitation system designed and developed based on FEM analysis proved to be economical due to its simplicity and has the added advan- tage of reduced harmonics in the output phase voltage.