Pharmacological inhibition of protein kinases in intact cells: antagonism of beta adrenergic receptor ligand binding by H-89 reveals limitations of usefulness

The use of pharmacological inhibitors of protein kinases represents a potentially powerful tool in dissecting the regulatory features of intracellular signaling pathways. However, although the in vitro potency, selectivity, and efficacy of numerous kinase inhibitors have been characterized, little is known regarding the usefulness of these compounds as inhibitors in intact cells. In attempting to characterize the role of protein kinase A (PKA) in regulating the beta-2 adrenergic receptor (AR) in human airway cells, we observed a seemingly profound capacity of the isoquinoline H-89, a potent and widely used PKA inhibitor, to attenuate agonist-mediated desensitization of the beta-2 AR. Although additional experiments identified H-89 as an effective inhibitor of intracellular PKA, extended analysis of the compound determined the principal effect of H-89 was via its action as a beta-2 AR antagonist. Pretreatment with or the acute addition of H-89 significantly attenuated isoproterenol-stimulated cAMP accumulation. In cells pretreated with H-89 and then washed extensively, the subsequent dose-dependent response to isoproterenol suggested beta-2 AR antagonism by retained H-89. Competition binding of [125I]iodopindolol established Ki values of approximately 180 nM and 350 nM for H-89 antagonism of beta-2 AR and beta-1 AR, respectively. Additional receptor binding studies suggest selectivity of H-89 for the beta-2 AR and beta-1 AR, although a weak antagonism (Ki values of approximately 10 microM or greater) of other G protein-coupled receptors was observed. Results from additional pharmacological and biochemical analyses of various protein kinase inhibitors further established the need for careful characterization of pharmacological inhibitors when used in intact cell models.     

Developing an Infrastructure to Support Communities of Learning

Through the National Grid for Learning (NGfL) the government is calling for new thinking about the potential of current educational institutions to support lifelong learning. One aspect of the reform is the creation and connection of a learning 'community'. However, knowledge of best practice for implementing reform will only come about through understanding the processes that occur in communities of learning, and, from this, developing an infrastructure to support these processes. The BT Laboratories (BTL) research project, 'HomeLearn', examined the impact of an on-line educational service on a community of parents, children and teachers. The creation of a home/school network was the first step in involving the wider community in the learning process. The three-month trial demonstrated that children were enthusiastic and creative in their use of communications technologies in the home. They were extremely confident communicating with adults over a network; they learnt to organise meetings, to manage their own time and to resolve conflicts. Some of the families integrated the information communications technologies into their everyday lives. They gradually built up knowledge of the technology though experience and through learning from other members of the family. From a service provider perspective we learnt that the critical concept in the development of a learning infrastructure is the support of 'relationships', a concept which entails more than simply providing a communications link or displaying static information. The widespread availability of new technologies means that people have an ever-increasing choice about how they want to communicate and with whom. An infrastructure to support learning must provide choice, be dynamic and be personalised. It is only by passing control of the system over to the users that a truly living system will exist.     

Theoretical and experimental study of the second-order polarizabilities of Schiff''s bases for nonlinear optical applications

A series of twenty-four Schiff's bases was synthesized and nonresonant static molecular second order polarizabilities (β) of these compounds were theoretically calculated and compared with experimental values. The computational method employed obtained: (a) values of polarization versus static electric fields using a semiempirical Hamiltonian; (b) all tensor elements of β by performing polynomial fits of the former data, within the finite-field approach. The experimental values were obtained using a modified electric field induced second harmonic generation (EFISH) experiment with 1,4-dioxane as the solvent. The measured quantities were the projection of β on μ (the permanent dipole moment), relative to MNA (2-methyl-4-nitroaniline). The correlation between the predicted static molecular quantities and their corresponding experimental values was 0.95 (based on a simple least-squares regression forced through the origin). A factor of 8.7 ± 0.3 was determined to be the adjustment parameter for Schiff's bases to account for the solvent and dispersion effects at the fundamental wavelength of 1064 nm.     

Lattice dynamics of colloidal crystals during photopolymerization of acrylic monomer matrix

The photoinitiated bulk polymerization process, which has been used recently in the manufacture of solid optical diffraction filters, is examined to understand the dynamics of both the crystalline colloidal arrays (CCA) and the host monomer species. Our analysis indicates that volume shrinkage of the monomer, changes in the dielectric properties of the monomer, and inhomogeneities of polymerization reaction rate across the dispersion during the polymerization process, are the major contributors for observed lattice compression and lattice disorder of the CCA of silica spheres in polymerized acrylic/methacrylic ester films. The effect of orientation of photocell with respect to the radiation source on Bragg diffraction of CCA indicated the presence of convective stirring in the thin fluid system during the photopolymerization that deleteriously affects the periodic array structures. To devise reproducible and more efficient optical filters, experimental methods to minimize or eliminate convective instabilities in monomeric dispersions during polymerization are suggested. © 1998 Chapman & Hall     

Low Loss Sintered Dielectric Resonators with HTS Thick Films

The combination of high-Qdielectric resonators and high-temperature superconducting (HTS) films offer many advantages in the area of cellular and satellite communications. The high cost of single crystal dielectrics and HTS thin films may be unattractive in certain applications. Superconducting thick films and polycrystalline ceramic dielectrics offer a high performance, low-cost alternative to high-Qthin film/single crystal dielectric resonators. The loss of polycrystalline ceramics of A1₂O₃, Ba(Mg_1/3Ta_2/3)O₃ (BMT), and Zr_0.875 Sn_0.25Ti0.875O₄ (ZTS) has been studied. Alumina, A1_2Oin3, has been studied as a model material for dielectric loss. Theory predicts that the loss in single crystal sapphire should follow aT ₅ dependence. However, at low temperatures the loss is dominated by extrinsic losses due to crystal imperfection, residual dopant atoms, dislocations, and other lattice defects and theT ₅ dependence does not hold. In polycrystalline alumina the intrinsic loss is immediately masked by these extrinsic losses, even at room temperature, and a simpleT dependence is observed. Results on polycrystalline alumina show that Q’s well in excess of 105 at 10 GHz and 77 K can be achieved in a design made compact by the use of a HTS thick film shield.     

Transmission of cancer from organ donors

One hundred and seventeen of 270 (43%) recipients of organs obtained from donors with malignancies had evidence of transmitted cancers. In 9 instances these were removed from renal allografts immediately prior to transplantation. Including these cases there were 45 recipients of organs in which a neoplasm involved the allograft, 6 others in whom adjacent structures were invaded, and another 66 patients who had distant metastases. Precautions to prevent cancer transmission include meticulous preoperative screening of donors, careful examination of all organs at the time of harvesting, biopsy of any suspicious lesions, and routine donor autopsy, if possible.     

Chronic illness, pain and health behaviors of community behavioral health clients.

Persons with comorbid medical disorders, along with substance use disorders and mental illness, present complex treatment needs that are seldom addressed. Chronic physical illness negatively affects treatment participation and retention, decreasing effectiveness. Studies documenting higher medical morbidity and mortality in such persons have long been available. Less is known about their health behaviors. Respondents (n = 418) at a community behavioral health center were surveyed for prevalence of illness, pain, health behaviors, and interest in lifestyle change. A total of 73% reported at least one chronic health problem, and nearly half rated their health between fair and very poor. Most reported at least one negative health indicator or behavior (e.g., smoking). Encouragingly, nearly 50% desired lifestyle changes including smoking cessation, exercise, and stress management. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Eastern Europe in the global microelectronics world

With the opening up of Eastern Europe to the West, new market opportunities abound, especially in the electronics industry for both East and West European firms alike. However, many barriers have yet to be overcome before these new opportunities can be transformed into business reality. This paper reviews the recent trends in East European microelectronics together with the current outlook and future growth potential.     

Direct analysis of sugar alcohol borate complexes in plant extracts by matrix-assisted laser desorption/ionization fourier transform mass spectrometry

Matrix-assisted laser desorption/ionization Fourier transform mass spectrometry (MALDI/FTMS), operating in the negative ion mode, is used to directly observe sugar alcohol borate complexes in a number of plant fractions. The method involves virtually no sample workup and, in the case of celery phloem sap, requires only 40 nL of sap to observe the borate complex. The isolation and characterization of such soluble borate complexes is important in understanding the distribution of boron in plants. The results show that the complexes are composed of two mannitol or sorbitol ligands (L) complexed to a single borate center (B). In some cases, the boron is complexed to non-alditol monosaccharides. Sustained off-resonance collision irradiation dissociation of the BL2- complex, where L is a mannitol, gives fragments that confirm the proposed structure. Complexes of larger oligosaccharides have also been successfully observed using MALDI/FTMS. Semiempirical molecular orbital calculations (AM1) of the mannitol BL2- complex show that the most favorable configuration is with carbons 3 and 4 of both mannitol residues complexed to the borate. This allows maximum interaction of the remaining hydroxyls with the borate center.