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Selected AbstractsMolecular homochirality and the parity-violating energy difference.CHIRALITY, Issue 2 2008A critique with new proposals Abstract Previous proposals for the origin of molecular homochirality, based on the effect of the weak neutral current (WNC) on enantiomers, and the amplification of the resultant parity-violating energy difference (PVED), are possibly flawed. The additive amplification of PVED in crystals and polymers ("Yamagata hypothesis") cannot lead to detectable levels of optical activity, the original theory apparently overestimating PVED by a factor equal to Avogadro's number. An alternative theory based on the irreversible and spontaneous evolution of a dynamically fluctuating system is apparently impractical. However, the nonlinear amplification of PVED via autocatalytic polymerization may be possible as indicated by a simplified physico-chemical approach. This may also occur during crystallization and melting, and form the basis of the second order asymmetric transformation. (Thus, reported differences in the melting points of enantiomers in several cases may well be real). Also, the preponderance of racemic compounds over conglomerates may be based on the destabilization of the conglomerate by the action of the WNC on the crystalline lattice. The WNC may also be involved in the anomalous scattering of X-rays, which possibly arises from their circular polarization: the current theory would need to be revised accordingly. Chirality, 2008. © 2007 Wiley-Liss, Inc. [source] Expression and characterization of active site mutants of hevamine, a chitinase from the rubber tree Hevea brasiliensisFEBS JOURNAL, Issue 3 2002Evert Bokma Hevamine is a chitinase from the rubber tree Hevea brasiliensis. Its active site contains Asp125, Glu127, and Tyr183, which interact with the ,1 sugar residue of the substrate. To investigate their role in catalysis, we have successfully expressed wild-type enzyme and mutants of these residues as inclusion bodies in Escherichia coli. After refolding and purification they were characterized by both structural and enzyme kinetic studies. Mutation of Tyr183 to phenylalanine produced an enzyme with a lower kcat and a slightly higher Km than the wild-type enzyme. Mutating Asp125 and Glu127 to alanine gave mutants with ,,2% residual activity. In contrast, the Asp125Asn mutant retained substantial activity, with an approximately twofold lower kcat and an approximately twofold higher Km than the wild-type enzyme. More interestingly, it showed activity to higher pH values than the other variants. The X-ray structure of the Asp125Ala/Glu127Ala double mutant soaked with chitotetraose shows that, compared with wild-type hevamine, the carbonyl oxygen atom of the N -acetyl group of the ,1 sugar residue has rotated away from the C1 atom of that residue. The combined structural and kinetic data show that Asp125,and Tyr183 contribute to catalysis by positioning the,carbonyl oxygen of the N -acetyl group near to the C1 atom. This allows the stabilization of a positively charged transient intermediate, in agreement with a previous proposal that the enzyme makes use of substrate-assisted catalysis. [source] Blockade by ferrous iron of Ca2+ influx through N -methyl- d -aspartate receptor channels in immature cultured rat cortical neuronsJOURNAL OF NEUROCHEMISTRY, Issue 1 2002Noritaka Nakamichi Abstract Rat cortical neurons cultured for 3 days in vitro were loaded with the fluorescent indicator fluo-3 for assessment of intracellular free calcium ion (Ca2+) concentrations with the aid of a confocal laser-scanning microscope. In the absence of added MgCl2, the addition of NMDA induced a rapid but sustained increase in the number of fluorescent neurons in a concentration-dependent manner at a concentration range of 1,100 µm with the increase by KCl being transient. The addition of FeCl2, but not FeCl3, markedly inhibited the increase by NMDA in a reversible manner at concentrations of 10,200 µm, without affecting that by KCl. Extensive analyses revealed clear differentiation between inhibitions by ferrous iron and other channel blockers known to date. The inhibition by FeCl2 was completely prevented by the addition of two different iron chelators. Exposure to NMDA alone did not lead to cell death in immature cultured neurons, however, while further addition of FeCl2 invariably induced neuronal cell death 24 h after exposure. These results give support to our previous proposal that NMDA receptor complex may contain a novel site sensitive to blockade by ferrous iron in rat brain. [source] Comparison of three diagrammatic keys for the quantification of late blight in tomato leavesPLANT PATHOLOGY, Issue 6 2009F. M. Corręa Three diagrammatic grading keys were designed for the assessment of the severity of late blight (caused by Phytophthora infestans) in tomato leaves. Simplified and broad keys considered, respectively, six (3, 12, 22, 40, 60 and 77%) and eight (3, 6, 12, 22, 40, 60, 77 and 90%) levels of disease severity, whilst a modified key based on a previous proposal for potato late blight considered six levels (1, 5, 10, 16, 32 and 50%). The keys were validated by 24 evaluators who assessed digital images of tomato leaves exhibiting different areas with lesions. Evaluator errors were compared using a mixed model in which evaluators were considered as random effects and the keys and evaluations as fixed effects. The accuracy and precision of the evaluators were compared by simple linear regression between the estimated and actual values of disease severity. The repeatability of evaluators was assessed using Pearson's correlation coefficient. There was significant (P , 0·001) variability amongst the errors made by evaluators, although the precision of each of the three keys was high with a coefficient of determination (R2) of 0·96, 0·93 and 0·83 for the simplified, broad and modified key, respectively. Repeatability of estimations amongst the evaluators was adequate (correlation coefficients of 0·91, 0·91 and 0·90 for the three keys, respectively). The simplified and broad keys resulted in higher precision and accuracy for the estimation of severity than did the modified key. Since the simplified key considers a smaller number of disease severity levels, its use is recommended in the assessment of late blight in tomato leaves. [source] Willow (Salix spp.) and aspen (Populus tremula) regrowth after felling by the Eurasian beaver (Castor fiber): implications for riparian woodland conservation in ScotlandAQUATIC CONSERVATION: MARINE AND FRESHWATER ECOSYSTEMS, Issue 1 2009Kevin Jones Abstract 1.A proposed trial reintroduction of the Eurasian beaver (Castor fiber L.) to Scotland has recently been approved (May 2008). A previous proposal was turned down by the licensing authority, partly over the perceived risks to woodland within a Special Area of Conservation. 2.This paper presents data on two years of willow (Salix spp.) and aspen (Populus tremula L.) regrowth following tree felling by captive beavers within two large semi-natural enclosures in eastern Scotland. Both willow and aspen are highly preferred by beavers. 3.Regrowth of willow stems was quantified from cut stumps, felled logs and partially severed (,incompletely felled') trees, and compared with growth of stems in the canopies of untouched willows. 4.Maximum annual regrowth was observed in incompletely felled trees, followed by cut stumps and then unfelled control trees. Mean regrowth rates were ca 18 m regrowth per metre of measured tree section per year in incompletely felled trees. This was approximately 12 times more annual regrowth than that observed in unfelled trees, despite apparently selective browsing of the regrowth of incompletely felled trees by roe deer. 5.Although incompletely felled willows represent a relatively small component of felling activity (9% of all felled willow), they contribute disproportionately to the biomass of beaver-engineered riparian woodlands, creating a more multi-layered habitat structure. 6.Willow regrowth in inundated areas was avoided by deer, and harvesting of regrowth by beavers was rare. 7.Significant regrowth from 11 stumps of aspen in 2 years post-felling occurred by suckering, with 85 suckers of mean height 2.1 m being recorded, all of which were subsequently avoided by browsing deer and beavers. 8.These data suggest that rapid regeneration of willow and aspen will occur in riparian woodlands in the event of major felling activity by Eurasian beaver, even in the presence of low to moderate levels of roe deer browsing, and that the conservation status of both these trees or the wider habitats that they form would not be threatened by a well planned and managed reintroduction of beavers to Scotland. Copyright © 2008 John Wiley & Sons, Ltd. [source] Evolution and persistence of the ciliumCYTOSKELETON, Issue 12 2007Peter Satir Abstract The origin of cilia, a fundamental eukaryotic organelle, not present in prokaryotes, poses many problems, including the origins of motility and sensory function, the origins of nine-fold symmetry, of basal bodies, and of transport and selective mechanisms involved in ciliogenesis. We propose the basis of ciliary origin to be a self-assembly RNA enveloped virus that contains unique tubulin and tektin precursors. The virus becomes the centriole and basal body, which would account for the self-assembly and self-replicative properties of these organelles, in contrast to previous proposals of spirochaete origin or endogenous differentiation, which do not readily account for the centriole or its properties. The viral envelope evolves into a sensory bud. The host cell supplies the transport machinery and molecular motors to construct the axoneme. Polymerization of cytoplasmic microtubules in the 9 + 0 axoneme completes the 9 + 2 pattern. Cell Motil. Cytoskeleton 2007. © 2007 Wiley-Liss, Inc. [source] Lystrosaurus species composition across the Permo,Triassic boundary in the Karoo Basin of South AfricaLETHAIA, Issue 2 2007JENNIFER BOTHA Lystrosaurus is one of the few therapsid genera that survived the end-Permian mass extinction, and the only genus to have done so in abundance. This study identifies which species of Lystrosaurus have been recovered from Permian and Triassic strata to determine changes in the species composition across the Permo,Triassic (P,T) boundary in the Karoo Basin of South Africa. Data generated from museum collections and recent fieldwork were used to stratigraphically arrange a total of 189 Lystrosaurus specimens to determine which species survived the extinction event. Results reveal that L. curvatus and L. maccaigi lived together on the Karoo floodplains immediately before the extinction event. L. maccaigi did not survive into the Triassic in South Africa. L. curvatus survived, but did not flourish and soon became extinct. Two new species of Lystrosaurus, L. murrayi and L. declivis, appeared in the Early Triassic. It is possible that L. murrayi and L. declivis occupied different niches to L. maccaigi and L. curvatus, and had special adaptations that were advantageous in an Early Triassic environment. We suggest that L. maccaigi may be used as a biostratigraphic marker to indicate latest Permian strata in South Africa and that, in support of previous proposals, the genus Lystrosaurus should not be used as a sole indicator of Triassic-aged strata. Our field data also show that L. curvatus may be regarded as a biostratigraphic indicator of the P,T boundary interval. [source] Formation of Trichlorinated Dibenzo- p -dioxins from 2,4-Dichlorophenol and 2,4,5-Trichlorophenolate: A Theoretical StudyCHEMPHYSCHEM, Issue 11 2006Ernesto Suárez Abstract The reaction of the 2,4,5-trichlorophenolate anion with 2,4-dichlorophenol to afford trichlorinated dibenzo- p -dioxins (T3CDDs) is investigated at the B3LYP/6-31+G(d) and B3LYP/6-311+G(3df,2p)//B3LYP/6-31+G(d)+ZPVE(B3LYP/6-31+G(d)) levels of theory. The first stage of the process corresponds to the formation of a predioxin, which can evolve through four different routes. Two of them lead directly to the products 2,3,7-T3CDD and 1,3,8-T3CDD, and the other two afford different predioxin-type intermediates, which in turn can evolve through all or some of the four routes to give new predioxins or T3CDD. Consequently, the theoretical results obtained show plainly the complex chemistry implied in the formation of dioxins from chlorophenols via anionic mechanisms by disclosing all the critical structures through which the system evolves, thus allowing assessment of the viability of the different mechanistic routes and the accessible products. The statistical thermodynamics treatment at 1 atm and 298.15, 600, 900, and 1200 K indicates that at higher temperatures, the Gibbs energy barrier for the formation of the initial predioxin is clearly the rate-determining step for the whole process, but at lower temperatures the Gibbs energy barrier for this step is similar to those for its evolution into 2,3,7-T3CDD. This result is in contrast with previous proposals that the closure of the central ring is the rate-limiting step. Finally, according to our results the rate constant for the formation of polychlorinated dibenzo- p -dioxins increases with the temperature, in agreement with the experimental observation that the conversion of trichlorophenols increases when going from 600 to 900 K in the gas phase in the absence of catalysts, and with DFT molecular dynamics results. [source] |