Substrate phosphorylation in the protein kinase Cgamma knockout mouse

The phosphorylation state of three identified neural-specific protein kinase C substrates (RC3, GAP-43/B-50, and MARCKS) was monitored in hippocampal slices of mice lacking the gamma-subtype of protein kinase C and wild-type controls by quantitative immunoprecipitation following 32Pi labeling. Depolarization with potassium, activation of glutamate receptors with glutamate, or direct stimulation of protein kinase C with a phorbol ester increased RC3 phosphorylation in wild-type animals but failed to affect RC3 phosphorylation in mice lacking the gamma-subtype of protein kinase C. Our results suggests the following biochemical pathway: activation of a postsynaptic (metabotropic) glutamate receptor stimulates the gamma-subtype of protein kinase C, which in turn phosphorylates RC3. The inability to increase RC3 phosphorylation in mice lacking the gamma-subtype of protein kinase C by membrane depolarization or glutamate receptor activation may contribute to the spatial learning deficits and impaired hippocampal LTP observed in these mice.     

Supplementation of squalene attenuates experimentally induced myocardial infarction in rats

We have investigated the preventive effects of squalene against isoprenaline-induced myocardial infarction in male albino rats. Supplementation with squalene significantly prevented the isoprenaline-induced adverse changes in the levels of protein and glycoprotein components in plasma and heart tissue of experimental groups of rats. It exerted an antioxidant effect by inhibiting the isoprenaline-induced lipid peroxidation and by maintaining the level of non-enzymatic free radical-scavenger, reduced glutathione at near normalcy. Histopathological observations also confirmed the possible cardioprotective action of squalene by maintaining the normal architecture of the heart tissue. The results of the present investigation demonstrate that supplementation with squalene offers cardioprotection in experimental rats by its antioxidant and membrane- stabilizing properties.     

Heavy metal concentrations in fish,shellfish and fish products from internal markets of India vis-a-vis international standards

Heavy metals are an important group of chemical contaminants and food is the major vehicle for entry into the system. Fish constitute a major source of heavy metals in food. Concentration of heavy metals in commercially important species of fish, shellfish and fish products from fish markets in and around the Cochin area was evaluated using an atomic absorption spectrometer. The concentration ranges of Cd, Pb, Hg, Cr, As, Zn, Cu, Co, Mn, Ni, and Se in the samples were <0.07–1, <0.07–1.32, <0.05–2.31, <0.05 to 3.65, <0.1–4.14, 0.6 to 165, 0.15 to 24, <0.02 to 0.85, <0.08 to 9.2, <0.032–1.38 and; <0.03–1.35 mg/kg, respectively. The present study showed that different metals were present in the samples at different levels but within the maximum residual levels prescribed by the EU and USFDA and the fish and shellfish from these areas, in general, are safe for human consumption.     

Cover Feature: Molecular Highway Patrol for Ribosome Collisions (ChemBioChem 20/2023)

During translation, messenger RNAs (mRNAs) are decoded by ribosomes which can stall for various reasons. These include chemical damage, codon composition, starvation, or translation inhibition. Trailing ribosomes can collide with stalled ribosomes, potentially leading to dysfunctional or toxic proteins. Such aberrant proteins can form aggregates and favor diseases, especially neurodegeneration. To prevent this, both eukaryotes and bacteria have evolved different pathways to remove faulty nascent peptides, mRNAs and defective ribosomes from the collided complex. In eukaryotes, ubiquitin ligases play central roles in triggering downstream responses and several complexes have been characterized that split affected ribosomes and facilitate degradation of the various components. As collided ribosomes signal translation stress to affected cells, in eukaryotes additional stress response pathways are triggered when collisions are sensed. These pathways inhibit translation and modulate cell survival and immune responses. Here, we summarize the current state of knowledge about rescue and stress response pathways triggered by ribosome collisions.     

Changes in Effective Thermal Conductivity During the Carbothermic Reduction of Magnetite Using Graphite

Knowledge of the effective thermal diffusivity changes of systems undergoing reactions where heat transfer plays an important role in the reaction kinetics is essential for process understanding and control. Carbothermic reduction process of magnetite containing composites is a typical example of such systems. The reduction process in this case is highly endothermic and hence, the overall rate of the reaction is greatly influenced by the heat transfer through composite compact. Using Laser-Flash method, the change of effective thermal diffusivity of magnetite-graphite composite pellet was monitored in the dynamic mode over a pre-defined thermal cycle (heating at the rate of 7 K/min to 1423 K (1150 °C), holding the sample for 270 minutes at this temperature and then cooling it down to the room temperature at the same rate as heating). These measurements were supplemented by Thermogravimetric Analysis under comparable experimental conditions as well as quenching tests of the samples in order to combine the impact of various factors such as sample dilatations and changes in apparent density on the progress of the reaction. The present results show that monitoring thermal diffusivity changes during the course of reduction would be a very useful tool in a total understanding of the underlying physicochemical phenomena. At the end, effort is made to estimate the apparent thermal conductivity values based on the measured thermal diffusivity and dilatations.     

Determination of interface-state parameters in a MOS capacitor by DLTS

A modification of the DLTS (Deep Level Transient Spectroscopy) technique for interface-state measurement is described in which the surface potential is used to determine the energy of the interface states contributing to the emission signal. This technique allows an accurate and unambiguous determination of interface-state energies and cross sections. Expressions are determined for interface-state emission as a function of surface potential. Measurements of interface-state density and majority-carrier cross sections as functions of energy for n- and p-type MOS samples are presented.     

M.O.S. switched-capacitor amplifiers

Switched-capacitor voltage gain elements, in which the gain is precisely controlled by the ratio of two capacitors, are presented. The voltage gain is obtained by means of a transconductance element operating into a switched-capacitor resistor. Circuit configurations use a single operational amplifier or a unity-gain buffer. In some of these configurations, the offset voltage of the operational amplifier or the buffer is not amplified.     

The conserved arginine in rho-GTPase-activating protein is essential for efficient catalysis but not for complex formation with Rho.GDP and aluminum fluoride

The Rho family of small GTP-binding proteins are downregulated by an intrinsic GTPase, which is enhanced by GTPase-activating proteins (GAPs). RhoGAPs contain a single conserved arginine residue that has been proposed to be involved in catalysis. Here, the role of this arginine has been elucidated by mutagenesis followed by determination of catalytic and equilibrium binding constants using single-turnover kinetics, isothermal titration calorimetry, and scintillation proximity assays. The turnover numbers for wild-type, R282A, and R282K RhoGAPs were 5.4, 0.023, and 0.010 s-1, respectively. Thus, the function of this arginine could not be replaced by lysine or alanine. Nevertheless, the R282A mutation had a minimal effect on the binding affinity of RhoGAP for either Rho. GTP or Rho.GMPPNP, which confirms the importance of the arginine residue for catalysis as opposed to formation of the protein-protein complex. The R282A mutant RhoGAP still increased the hydrolysis rate of Rho.GTP by 160-fold, whereas the wild-type enzyme increased it by 38000-fold. We conclude that this arginine contributes half of the total reduction of activation energy of catalysis. In the presence of aluminum fluoride, the R282A mutant RhoGAP binds almost as well as the wild type to Rho.GDP, demonstrating that the conserved arginine is not required for this interaction. The affinity of wild-type RhoGAP for the triphosphate form of Rho is similar to that for Rho.GDP with aluminum fluoride. These last two observations show that this complex is not associated with the free energy changes expected for the transition state, although the Rho.GDP.AlF4-.RhoGAP complex might well be a close structural approximation.