Biosynthesis of growth hormone-releasing factor by fetal rat cerebrocortical and hypothalamic cells

The biosynthesis of growth hormone-releasing factor (GRF) by cerebrocortical tissue is controversial. Although several reports have indicated its presence in certain rat cortical areas and in cultured rat hypothalamic cells, no data exist demonstrating its biosynthesis in these areas. In this study, we have investigated the capacity of fetal rat cerebrocortical and hypothalamic cells in culture for synthesizing GRF. Fetal cerebrocortical and hypothalamic cells were exposed to [3H]Arg for 48 h. Medium and cell extracts were processed and [3H]Arg-IR-rGRF was isolated by affinity chromatography and characterized by HPLC. Intracellular [3H]Arg-IR-rGRF from both hypothalamic and cerebrocortical cells exhibited four major peaks, one of them coeluting with synthetic rGRF. In cerebrocortical cultures, newly synthesized and released [3H]Arg-IR-rGRF showed a similar pattern to the cell content. However, in media from hypothalamic cells, higher hydrophobicity molecular forms were absent. The data demonstrated that fetal cerebrocortical and hypothalamic cells in primary culture synthesize GRF with similar posttranslational processing, but with different molecular patterns of secretion.     

Influence of mechanical properties of the sheet material on formability in single point incremental forming

New trends in sheet metal forming are rapidly developing and several new forming processes have been proposed to accomplish the goals of flexibility and cost reduction. Among them single point incremental forming operations, in which the final shape of the component is obtained by the relative movement of a simple and small punch with respect to the blank, appear quite promising.In the paper, material formability issues in incremental forming were studied. Some relevant correlations among material formability and other mechanical properties of the material were analysed. The FLD₀ value, i.e. the major strain at fracture in plane strain conditions, was determined for different materials and the influence of the main material parameters on formability was accurately investigated through a statistical analysis.     

The catalytic domain of protein kinase C chimeras modulates the affinity and targeting of phorbol ester-induced translocation

Emerging evidence suggests important differences among protein kinase C (PKC) isozymes in terms of their regulation and biological functions. PKC is regulated by multiple interdependent mechanisms, including enzymatic activation, translocation of the enzyme in response to activation, phosphorylation, and proteolysis. As part of our ongoing studies to define the factors contributing to the specificity of PKC isozymes, we prepared chimeras between the catalytic and regulatory domains of PKCalpha, -delta, and -epsilon. These chimeras, which preserve the overall structure of the native PKC enzymes, were stably expressed in NIH 3T3 fibroblasts. Their intracellular distribution was similar to that of the endogenous enzymes, and they responded with translocation upon treatment with phorbol 12-myristate 13-acetate (PMA). We found that the potency of PMA for translocation of the PKCalpha/x chimeras from the soluble fraction was influenced by the catalytic domain. The ED50 for translocation of PKCalpha/alpha was 26 nM, in marked contrast to the ED50 of 0.9 nM in the case of the PKCalpha/epsilon chimera. In addition to this increase in potency, the site of translocation was also changed; the PKCalpha/epsilon chimera translocated mainly into the cytoskeletal fraction. PKCx/epsilon chimeras displayed twin isoforms with different mobilities on Western blots. PMA treatment increased the proportion of the higher mobility isoform. The two PKCx/epsilon isoforms differed in their localization; moreover, their localization pattern depended on the regulatory domain. Our results emphasize the complex contributions of the regulatory and catalytic domains to the overall behavior of PKC.     

Thermophiles as Potential Source of Novel Endotoxin Antagonists: the Full Structure and Bioactivity of theLipo‐oligosaccharide from Thermomonas hydrothermalis

Thermomonas hydrothermalis is a Gram‐negative thermophilic bacterium that is able to live at 50 °C. This ability is attributed to chemical modifications, involving those to bacterial cell‐wall components, such as proteins and (glyco)lipids. As the main component of the outer membrane of Gram‐negative bacteria, lipopolysaccharides (LPSs) are exposed to the environment, thus they can undergo structural chemical changes to allow thermophilic bacteria to live at their optimal growth temperature. Furthermore, as one of the major target of the eukaryotic innate immune system, LPS elicits host immune response in a structure‐dependent mode; thus the uncommon chemical features of thermophilic bacterial LPSs might exert a different biological action on the innate immune system—an antagonistic effect, as shown in studies of LPS structure–activity relationship in the ongoing research into antagonist LPS candidates. Here, we report the complete structural and biological activity analysis of the lipo‐oligosaccharide isolated from Thermomonas hydrothermalis, achieved by a multidisciplinary approach (chemical analysis, NMR, MALDI MS and cellular immunology). We demonstrate a tricky and interesting structure combined with a very interesting effect on human innate immunity.     

A design methodology for high-order class-D audio amplifiers

This paper presents a design methodology for high-order class-D amplifiers, based on their similarity with sigma–delta ( $\Upsigma\Updelta$ ) modulators, for which established theory and toolboxes are available. The proposed methodology, which covers the entire design flow, from specifications to component sizing, is validated with three design examples, namely a second-order, a third-order, and a fourth-order class-D amplifier. Moreover, the third-order class-D amplifier has been integrated on silicon and characterized, further confirming the validity of the whole design flow. The achieved results demonstrate that high-order class-D amplifiers can achieve total-harmonic-distortion (THD) performance compatible with the specifications of high-end audio applications (THD  ≈ 90 dB), which would be unfeasible with conventional first-order class-D amplifiers.     

A New Functional Form for Equations of State for Some Weakly Associating Fluids

A new functional form for equations of state for polar and weakly associating fluids was developed. It was established with a simultaneous optimization algorithm developed previously. As a result, equations of state in terms of the Helmholtz energy as a function of temperature and density were developed for hydrogen chloride (HCl) valid within \(T =\) (155–330) K and pressures up to \(p = 20\) MPa, diethyl ether (DEE) valid within \(T =\) (270–500) K and pressures up to \(p = 40\) MPa, and methyl chloride (R40) valid within \(T =\) (230–630) K and pressures up to \(p =\) 100 MPa. Those equations can be used for the calculation of all thermodynamic properties, including density, internal energy, enthalpy, heat capacity, speed of sound, saturation properties, etc.     

Grafting Single Molecule Magnets on Gold Nanoparticles

The chemical synthesis and characterization of the first hybrid material composed by gold nanoparticles and single molecule magnets (SMMs) are described. Gold nanoparticles are functionalized via ligand exchange using a tetrairon(III) SMM containing two 1,2‐dithiolane end groups. The grafting is evidenced by the shift of the plasmon resonance peak recorded with a UV–vis spectrometer, by the suppression of nuclear magnetic resonance signals, by X‐ray photoemission spectroscopy peaks, and by transmission electron microscopy images. The latter evidence the formation of aggregates of nanoparticles as a consequence of the cross‐linking ability of Fe₄ through the two 1,2‐dithiolane rings located on opposite sides of the metal core. The presence of intact Fe₄ molecules is directly proven by synchrotron‐based X‐ray absorption spectroscopy and X‐ray magnetic circular dichroism spectroscopy, while a detailed magnetic characterization, obtained using electron paramagnetic resonance and alternating‐current susceptibility, confirms the persistence of SMM behavior in this new hybrid nanostructure.