Reciprocal regulation of neu tyrosine kinase activity and caveolin-1 protein expression in vitro and in vivo. Implications for human breast cancer

Neu (c-erbB2) is a proto-oncogene product that encodes an epidermal growth factor-like receptor tyrosine kinase. Amplification of wild-type c-Neu and mutational activation of Neu (Neu T) have been implicated in oncogenic transformation of cultured fibroblasts and mammary tumorigenesis in vivo. Here, we examine the relationship between Neu tyrosine kinase activity and caveolin-1 protein expression in vitro and in vivo. Recent studies have suggested that caveolins may function as negative regulators of signal transduction. Our current results show that mutational activation of c-Neu down-regulates caveolin-1 protein expression, but not caveolin-2, in cultured NIH 3T3 and Rat 1 cells. Conversely, recombinant overexpression of caveolin-1 blocks Neu-mediated signal transduction in vivo. These results suggest a reciprocal relationship between c-Neu tyrosine kinase activity and caveolin-1 protein expression. We next analyzed a variety of caveolin-1 deletion mutants to map this caveolin-1-dependent inhibitory activity to a given region of the caveolin-1 molecule. Results from this mutational analysis show that this functional in vivo inhibitory activity is contained within caveolin-1 residues 32-95. In accordance with these in vivo studies, a 20-amino acid peptide derived from this region (the caveolin-1 scaffolding domain) was sufficient to inhibit Neu autophosphorylation in an in vitro kinase assay. To further confirm or refute the relevance of our findings in vivo, we next examined the expression levels of caveolin-1 in mammary tumors derived from c-Neu transgenic mice. Our results indicate that dramatic reduction of caveolin-1 expression occurs in mammary tumors derived from c-Neu-expressing transgenic mice and other transgenic mice expressing downstream effectors of Neu-mediated signal transduction, such as Src and Ras. Taken together, our data suggest that a novel form of reciprocal negative regulation exists between c-Neu and caveolin-1.     

Effects of neuromuscular blocking agents on excitatory transmission and gamma-aminobutyric acidA-mediated inhibition in the rat hippocampal slice

BACKGROUND: Although neuromuscular blocking agents do not cross the blood-brain barrier, they may penetrate the central nervous system under particular circumstances and eventually cause neurotoxic consequences. METHODS: The effects of neuromuscular blocking agents on excitatory and inhibitory transmission in area CA1 of rat hippocampal slices were investigated using extracellular and intracellular recording techniques. RESULTS: Application of atracurium in the perfusion medium resulted in a dose-dependent enhancement of excitatory synaptic responses averaging 48.7 +/- 4.3% at a concentration of 10 nM. This effect was correlated with an increase in the size of the presynaptic fiber volley. Laudanosine, but not pancuronium bromide or vecuronium bromide, produced similar changes. In addition, atracurium and laudanosine blocked inhibitory transmission and reduced intracellularly recorded gamma-aminobutyric acidA receptor-mediated potentials. These effects were observed only at concentrations >1 microM and were not reproduced by pancuronium bromide and vecuronium bromide. CONCLUSIONS: Atracurium and its metabolite, laudanosine, contrary to pancuronium bromide and vecuronium bromide, produce two distinct effects on hippocampal slices. They enhance excitatory transmission and neuronal excitability and they block inhibitory gamma-aminobutyric acidA-mediated synaptic responses.     

Thermal stability of polyacrylamide solutions: effect of residual impurities in the molecular-weight-degradation process upon heating

Summary We have investigated the effect of temperature on the chemical (hydrolysis) and thermal (molecular weight) stability of purified polyacrylamide solution.From viscosity and light scattering data we show that no polymer degradation due to oxygen contamination results from heating polymer solutions although higher temperature is responsible for hydrolysis of amide groups.     

MITOM: a new unfolding code based on a spectra model method applied to neutron spectrometry

The MITOM code was developed at UAB (Universitat Autònoma de Barcelona) for unfolding neutron spectrometric measurements with a Bonner spheres system (BSS). One of the main characteristics of this code is that an initial parameterisation of the neutron energy components (thermal, intermediate and fast) is needed. This code uses the Monte Carlo method and the Bayesian theorem to obtain a set of solutions achieving different criteria and conditions between calculated and measured count rates. The final solution is an average of the acceptable solutions. The MITOM code was tested for ISO sources and a good agreement was observed between the reference values and the unfolded ones for global magnitudes. The code was applied recently to characterise both thermal SIGMA and CANEL/T400 sources of the IRSN facilities. The results of these applications were very satisfactory as well.     

Prediction of work piece geometry in electrochemical cavity sinking

A computer-implemented model for predicting ECM work piece geometry has been developed and experimentally verified with a commercial ECM machine for cavity sinking in copper and 302-stainless steel with 2N K NO₃ electrolyte. Constant tool piece feed rates of 7–10 × 10^?4 cm/s, and applied voltages of 11–25 V were used. The model predicts the dependence of work piece geometry on operating conditions and on the electrochemical and physical properties of the metal—electrolyte pair. Comparison of eight equilibrium and six unsteady state experimental cavity profiles in copper showed satisfactory agreement with predictions, as did five equilibrium profiles for cavity sinking in 302-stainless steel.     

Velocity of surface carriers in inversion layers on silicon

Velocity-field curves for surface free-carriers in silicon are determined from measurements on resistivegate IGFETs. The measurements were performed on n-channel devices fabricated on both (100) and (111) substrates and on p-channel devices fabricated on (100) substrates. The channel length of the devices is ～8 μm and the impurity concentration of the substrates is ～ 10¹⁵ cm^?3. The dependence of velocity on the field strength along the channel is found to be well approximated by an empirical relationship involving three parameters: low-field mobility μ₀, a critical field $\text{E}{}_{\mathrm{cy}}$ signalling the onset of velocity saturation, and a parameter α that determines the curvature between the constant-mobility and constant-velocity branches of the curve. The curve-fitting parameters are given in tabular form for the two n-channel and one p-channel systems studied. The dependence of the velocity-field curves on temperatures in the range 100–350K is also reported.     

Pasture intake and substitution rate effects on nutrient digestion and nitrogen metabolism during continuous culture fermentation

A continuous culture system was used to investigate ruminal digestion in response to increased pasture intake and three different substitution rates (SR) in a 4 x 4 Latin square design. The treatments were 1) low pasture (55 g dry matter (DM)/d, 2) medium pasture (MP, 65 g DM/d), 3) high pasture (75 g DM/d), and 4) pasture (45 g DM/d) plus concentrate (PC, 30 g DM/d). Treatments were designed to produce a low (0.33), medium (0.67), and high (1.00) SR (g of pasture/g of concentrate) by contrasting the low, medium, and high pasture intake treatments with the pasture plus concentrate treatment, respectively. Pasture was fed at 0630, 1000, 1730, and 2100 h, and concentrate at 0600 and 1700 h. Digestibility of DM and neutral detergent fiber were not affected by the amount of pasture. As the amount of pasture increased, pH decreased linearly, and total volatile fatty acid and NH3-N concentrations, and nonammonia N and bacterial N flows increased linearly. Concentrate supplementation did not affect DM digestibility at high SR but increased DM digestibility at low SR. Concentrate supplementation reduced pH and NH3-N concentrations at the three SR. Concentrate supplementation reduced the ratio of rumen degradable N to rumen degradable organic matter; however, the mechanism depended on the SR. High SR, concentrate supplementation reduced rumen degradable N, which reduced NIH-N concentration without affecting bacterial N flow. At low SR, concentrate supplementation increased rumen degradable organic matter, which reduced NH3-N concentration and increased bacterial N flow. Based on these results, at low SR, concentrate supplementation may enhance animal performance because of higher total DM intake and synthesis of microbial protein.     

Supplemental carbohydrate sources for lactating dairy cows on pasture

Two experiments were conducted to evaluate steam-flaked corn and nonforage fiber sources as supplemental carbohydrates for lactating dairy cows on pasture. Cows were allotted to a new paddock of an orchardgrass (Dactylis glomerata L.) pasture twice daily in one group in both trials. In experiment 1, 28 Holstein cows, averaging 216 d in milk, were randomly assigned to either a cracked-corn (CC) or a steam-flaked (SFC) supplement in a split plot design. The supplement contained 66.7% of corn and a protein/mineral pellet. In experiment 2, 28 Holstein cows, averaging 182 d in milk, were randomly assigned to either a ground corn (GC) or a nonforage fiber (NFF)-based supplemented in a single reversal design. The GC supplement contained 85% ground corn plus protein, mineral, and vitamins. The NFF supplement contained 35% ground corn, 18% beet pulp, 18% soyhulls, 8% wheat middlings plus protein, mineral, and vitamins. In both experiments, cows were fed the grain supplement twice daily after each milking at 1 kg/4 kg milk. In experiment 1, milk production (24.3 kg/d) and composition did not differ between treatments; however, plasma and milk urea N were lower with the SFC supplement. In experiment 2, milk production (27.5 kg/d) was not affected by treatments, which may be related to the medium quality of pasture grazed. The GC supplement tended to reduce plasma and milk urea N and increased milk protein percentage (3.23 vs. 3.19%). Pasture dry matter intake, measured using Cr2O3, did not differ between treatments in either experiment 1 (15.1 kg/d) or experiment 2 (12.2 kg/d). Milk production did not differ when mid-late lactation cows on pasture were supplemented with SFC or NFF instead of dry corn.     

Investigation of the morphotropic region in the Sr-doped lead hafnate-titanate solid solution Pb0.94Sr0.06Hf1-TixO3

Sr-doped lead hafnate titanate (PHT) oxides were synthesized by solid-state reaction and carefully studied in order to identify the reaction mechanisms. The reaction sequences in the formation of Sr-doped PbHfO₃, Sr-doped PbTiO}_{3 and Sr-doped PbHf_{1 - x}Ti_xO₃ were investigated using X-ray powder diffraction and differential thermal analysis. The Sr-doped PbHfO₃ oxide is mainly elaborated from the reaction between PbO and HfO₂, while Sr-doped PbTiO₃ oxide is formed from the reaction between Pb₃O₄ and TiO₂. The Sr-doped PbHf_1-xTi_xO₃ oxide is obtained from a solid-state reaction between Sr-doped PbHfO₃ and Sr-doped PbTiO₃ solid solutions. For x close to 0.5, tetragonal and rhombohedral ferroelectric phases coexist in a region commonly called the morphotropic region which presents the best dielectric and electromechanical properties. By means of X-ray powder diffraction, the boundaries of the morphotropic region in the (Pb_0.94Sr_0.06)Hf_1-xTi_xO₃ system have been determined for x ranging between 0.42 and 0.58. Finally, electrical and piezoelectric characterizations have been performed on Sr-doped PHT ceramics crystallizing in the morphotropic region. The compositional dependence of the electromechanical coupling factor (k_P), piezoelectric coefficients (d_{33 and g₃₃) and relative permittivity (_r) has been studied. An optimum composition has been determined as x equal to 0.51. © 2001 Kluwer Academic Publishers