Molecular pharmacology of voltage-dependent calcium channels

Voltage-dependent Ca2+ channels serve as the only link to transduce membrane depolarization into cellular Ca(2+)-dependent reactions. A wide variety of chemical substances that have the ability to modulate Ca2+ channels have been demonstrated both for their clinic utility and for importance in elucidating the molecular basis of various biological responses. Recently, introduction of molecular biology to pharmacology has brought a great deal of information about the molecular basis of drug action in Ca2+ channels. In this review, we attempt to overview recent progress in understanding the interactions between Ca2+ channels and their blockers, namely Ca2+ antagonists, from a molecular and structural point of view.     

Assessment of conformational parameters as predictors of limited proteolytic sites in native protein structures

Despite the importance of limited proteolysis in biological systems it isoften difficult to rationalize why a proteinase hydrolyses a particularbond, given a simple sequence specificity alone. Understanding of thestructural properties limiting the proteolysis represents a first step onthe pathway to control and manipulation of this phenomena. An expanded setof nick-sites in proteins of known tertiary structure, cut by both narrowand broad specificity proteinases, has been generated yielding a robustdata set of strictly limited sites. A critical evaluation of an expandedset of conformational parameters revealed a strong correlation with limitedproteolytic sites, although they are only modest predictors in isolation.The overall predictive power is significantly improved when theconformational parameters are combined in a weighted predictive scheme thatpermits their relative importance to be compared via a Metropolis searchprotocol. A subset of the parameters performs equally well demonstratingthe key determinants of susceptibility. The derived predictive algorithmhas been made available via the internet. Its utility for predicting othersurface-correlated features is also discussed.     

MyoD and Myf-5 differentially regulate the development of limb versus trunk skeletal muscle

The myogenic progenitors of epaxial (paraspinal and intercostal) and hypaxial (limb and abdominal wall) musculature are believed to originate in dorsal-medial and ventral-lateral domains, respectively, of the developing somite. To investigate the hypothesis that Myf-5 and MyoD have different roles in the development of epaxial and hypaxial musculature, we further characterized myogenesis in Myf-5- and MyoD-deficient embryos by several approaches. We examined expression of a MyoD-lacZ transgene in Myf-5 and MyoD mutant embryos to characterize the temporal-spatial patterns of myogenesis in mutant embryos. In addition, we performed immunohistochemistry on sectioned Myf-5 and MyoD mutant embryos with antibodies reactive with desmin, nestin, myosin heavy chain, sarcomeric actin, Myf-5, MyoD and myogenin. While MyoD(-/-) embryos displayed normal development of paraspinal and intercostal muscles in the body proper, muscle development in limb buds and brachial arches was delayed by about 2.5 days. By contrast, Myf-5(-/-) embryos displayed normal muscle development in limb buds and brachial arches, and markedly delayed development of paraspinal and intercostal muscles. Although MyoD mutant embryos exhibited delayed development of limb musculature, normal migration of Pax-3-expressing cells into the limb buds and normal subsequent induction of Myf-5 in myogenic precursors was observed. These results suggest that Myf-5 expression in the limb is insufficient for the normal progression of myogenic development. Taken together, these observations strongly support the hypothesis that Myf-5 and MyoD play unique roles in the development of epaxial and hypaxial muscle, respectively.     

Oxidative dehydrogenation of isobutane on phosphorous-modified graphitic mesoporous carbon

Graphitic mesoporous carbon was modified with phosphorous heteroatoms in order to tune the catalytic selectivity and to investigate the roles of different oxygen species for the oxidative dehydrogenation reaction of isobutane to isobutene. Small changes in the isobutane apparent activation energy are consistent with the notion that the phosphorous groups do not change the nature of the active sites but they interfere with the availability of the sites. Our results show that the improvement on selectivity is not proportional to the amount of phosphorous added. Small phosphorous content improved the selectivity by suppressing the combustion of isobutane. However, a higher amount of phosphorous groups lead to coverage of selective quinone sites and/or creation of active sites favorable to total oxidation.     

Performance analysis of MSP: feature-rich high-speed transportprotocol

A performance analysis of the MultiStream Protocol (MSP), a feature-rich flexible transport protocol designed to meet the needs of high-performance applications, is presented. The analysis presents the advantages and disadvantages of the implementation of several different protocol mechanisms, such as packet or block mode data transfer. It is based on the processing complexity associated with executing each protocol function. Results show that a feature-rich transport protocol can be implemented in which the protocol control processing overhead is not prohibitive to providing high performance to an application. Instead, the benefits of a lean protocol are achieved because the protocol services better fit application requirements. Also, the primary bottleneck for data transfer is shown to be the complexity associated with error control, which can reduced through the technique of transmitting packets in blocks     

High-performance liquid chromatography with light-scattering detection and desorption chemical-ionization tandem mass spectrometry of milk fat triacylglycerols

The utility of reverse-phase high-performance liquid chromatography (HPLC), desorption chemical-ionization mass spectrometry (DCI-MS) and tandem mass spectrometry (MS/MS) for the characterization of triacylglycerols in complex mixtures has been further explored. Triacylglycerols of anhydrous bovine milk fat were separated by using two reversephase C₁₈ HPLC columns, and eluents were monitored with an evaporative light-scattering detector. Fifty-eight fractions were resolved and analyzed by positive ion isobutane DCI-MS. The formation of protonated molecules and of major fragments corresponding to the random loss of any one of the constituent fatty acids readily identified acyl carbon numbers and the number of double bonds within each fatty acid. MS/MS was only required when the original mass spectra indicated the presence of more than one triacylglycerol or of impurities in a fraction. Protonated molecules produced by DCI were fragmented using high energy collisional activation, and the resulting ions were detected by MS/MS. Odd-chain triacylglycerols were also readily distinguished using this methodology. The positive ion DCI and MS/MS techniques described here demonstrate the usefulness of this approach for the characterization of triacylglycerols in complex mixtures.     

Fugitive Diffusion Barrier for Integration of Sol-Gel-Derived Lead Titanate with Oxidized Silicon Substrates

The crystalline phase development and microstructural changes with heating of sol-gel-derived lead titanate (PT) particles and films on silica with and without a fugitive (or removable) diffusion barrier layer were investigated. Amorphous gel-derived PT particles were deposited on SiO₂-coated TEM grids with and without polyimide (PI) or carbon barrier layers between SiO₂ and PT. TEM analysis showed that PI or carbon barriers prevented reaction between the gel-derived PT particles and SiO₂. PT particles crystallize and then the PI or carbon film decomposes. Sol-gel-derived PT films were deposited on oxidized Si substrates (Si/SiO₂) with and without a PI barrier layer. Perovskite PT films were prepared on Si/SiO₂ substrates with a PI barrier; however, some porosity remained in the films. Identically prepared films without the PI barrier formed a mixture of pyrochlore and perovskite. X-ray photoelectron spectroscopy results indicate that the PI film pre- vents the diffusion of Si into the PT film.     

The life cycle impact of refurbishment packages on residential buildings with different initial thermal conditions

Existing buildings constitute a large portion of the UK’s housing stock. Refurbishment of existing buildings can, therefore, have an important role in achieving the UK government’s CO₂ reduction targets. While building regulations and rating frameworks mainly focus on the improvements of the operational performance of buildings, Life Cycle Analysis is considered to be a more appropriate framework to account for long–term CO₂ savings. This study evaluates a range of retrofit approaches (simple, medium, and deep), in terms of Life Cycle Carbon Footprint applied on a terraced house—one of the most common housing archetypes in London. The initial state of the original building has also been examined assuming three initial states (never refurbished, refurbished in compliance with the 1976 and with the 2000 building regulations). Results showed that for all initial state scenarios, deep retrofit achieved the lowest life cycle carbon emissions, in absolute figures, compared to the simple and medium retrofits. Simple retrofit packages, on the other hand, achieved quick and significant improvements, especially in buildings with poor initial thermal conditions. The study also indicated that retrofit packages applied on highly efficient building fabrics result in longer carbon payback time periods. The study recommends establishing a ‘staggered’ retrofitting approach, which pushes for ‘older building first’ and ‘simple retrofit packages first’, as these gain quick CO₂ savings. Deep retrofit packages and treatment of relatively new buildings should be implemented at a later stage, to push buildings further to Zero–Carbon target.