Epicuticular wax variability of Juniperus deltoides R.P. Adams from the central Balkan – Ecology and chemophenetics

Juniperus deltoides R.P.Adams is a prickly, evergreen shrub that grows in the East Mediterranean on dry hills and mountains. Needle-like leaves have a thick cuticular layer on the surface, which serves as the ultimate barrier for prevention of water loss. n-Alkanes comprise the biggest part of this hydrophobic wax mixture. These compounds are studied as chemophenetic characters due to their chemical and physiological stability, and strong genetic control. Six central Balkan populations were studied and compared to one eastern and one western. Fifteen n-alkanes were identified, with C33 being dominant in all samples. Univariate and multivariate statistical analyses separated western population from all central Balkan populations, while the eastern Balkan population showed high similarity with the central ones. Climatic, orographic and pedological characteristics of the studied localities were also taken into account.     

How do reaction conditions affect the enantiopure synthesis of 2‐substituted‐1,4‐benzodioxane derivatives?

Several biologically active compounds structurally include the enantiopure 2‐substituted‐1,4‐benzodioxane scaffold. The straightforward racemization that affects reactions involving most of the common chemical reactives is thus a crucial issue. The developing of a completely stereo‐controlled synthetic route that does not affect the enantiomeric excess is consequently mandatory. It is also important to set up a reliable chiral HPLC method, able to follow the reaction, and to improve the synthetic performances. Here, we report the chiral investigation of two different synthons, we specifically evaluated the synthetic pathways that could be run in order to afford them, avoiding the racemization processes, which could normally occur in basic conditions. In addition, we developed peculiar chiral HPLC methods in order to resolve the enantiomers, define the enantiomeric excess, and fully characterize these compounds.     

No Matter – How?: Dealing with Matter‐less Stressors in LCA of Wind Energy Systems

The portfolio of impacts that are quantified in life cycle assessment (LCA) has grown to include rather different stressors than those that were the focus of early LCAs. Some of the newest life cycle impact assessment (LCIA) models are still in an early phase of development and have not yet been included in any LCA study. This is the case for sound emissions and noise impacts, which have been only recently modeled. Sound emissions are matter‐less, time dependent, and bound to the physical properties of waves. The way sound emissions and the relative noise impacts are modeled in LCA can show how new or existing matter‐less impacts can be addressed. In this study, we analyze, through the example of sound emissions, the specific features of a matter‐less impact that does not stem from the use of a kilogram of matter, nor is related to the emission of a kilogram of matter. We take as a case study the production of energy by means of wind turbines, contradicting the commonly held assumption that windmills have no emissions during use. We show how to account for sound emissions in the life cycle inventory phase of the life cycle of a wind turbine and then calculate the relative impacts using a noise LCIA model.     

Anti‐atherosclerotic effects of vitamin E – myth or reality?

Atherosclerosis and its complications such as coronary heart disease, myocardial infarction and stroke are the leading causes of death in the developed world. High blood pressure, diabetes, smoking and a diet high in cholesterol and lipids clearly increase the likelihood of premature atherosclerosis, albeit other factors, such as the individual genetic makeup, may play an additional role. Several epidemiological studies and intervention trials have been performed with vitamin E, and some of them showed that it prevents atherosclerosis. For a long time, vitamin E was assumed to act by decreasing the oxidation of LDL, a key step in atherosclerosis initiation. However, at the cellular level, vitamin E acts by inhibition of smooth muscle cell proliferation, platelet aggregation, monocyte adhesion, oxLDL uptake and cytokine production, all reactions implied in the progression of atherosclerosis. Recent research revealed that these effects are not the result of the antioxidant activity of vitamin E, but rather of precise molecular actions of this compound. It is assumed that specific interactions of vitamin E with enzymes and proteins are at the basis of its non-antioxidant effects. Vitamin E influences the activity of several enzymes (e.g. PKC, PP2A, COX-2, 5-lipooxygenase, nitric oxide synthase, NADPH-oxidase, superoxide dismutase, phopholipase A2) and modulates the expression of genes that are involved in atherosclerosis (e.g. scavenger receptors, integrins, selectins, cytokines, cyclins). These interactions promise to reveal the biological properties of vitamin E and allow designing better strategies for the protection against atherosclerosis progression.     

How Many Sirtuin Genes Are Out There? Evolution of Sirtuin Genes in Vertebrates With a Description of a New Family Member

Studying the evolutionary history of gene families is a challenging and exciting task with a wide range of implications. In addition to exploring fundamental questions about the origin and evolution of genes, disentangling their evolution is also critical to those who do functional/structural studies to allow a deeper and more precise interpretation of their results in an evolutionary context. The sirtuin gene family is a group of genes that are involved in a variety of biological functions mostly related to aging. Their duplicative history is an open question, as well as the definition of the repertoire of sirtuin genes among vertebrates. Our results show a well-resolved phylogeny that represents an improvement in our understanding of the duplicative history of the sirtuin gene family. We identified a new sirtuin gene family member (SIRT3.2) that was apparently lost in the last common ancestor of amniotes but retained in all other groups of jawed vertebrates. According to our experimental analyses, elephant shark SIRT3.2 protein is located in mitochondria, the overexpression of which leads to an increase in cellular levels of ATP. Moreover, in vitro analysis demonstrated that it has deacetylase activity being modulated in a similar way to mammalian SIRT3. Our results indicate that there are at least eight sirtuin paralogs among vertebrates and that all of them can be traced back to the last common ancestor of the group that existed between 676 and 615 millions of years ago.     

Who Is the King? The α‐Hydroxy‐β‐oxo‐α,β‐enone Moiety or the Catechol B Ring: Relationship between the Structure of Quercetin Derivatives and Their Pro‐Oxidative Abilities

Quercetin and other flavonoids have been reported to exhibit both antioxidant and pro‐oxidant properties. Most studies about the pro‐oxidative ability were conducted in the presence of metal ions, and the essential functional moiety of quercetin responsible for the pro‐oxidative effect is still unclear. In this study, we evaluated the pro‐oxidative abilities in the absence of metal ions of two quercetin derivatives, i.e., quercetin‐3′‐O‐β‐D ‐glucoside ( 1 ) and quercetin‐3‐O‐β‐D ‐glucoside ( 2 ), by assessing DNA cleavage and HO^.‐radical production. The binding mode between these compounds and DNA was studied by fluorescence and viscometric titrations. The results showed that 1 can efficiently induce oxidative damage to plasmid DNA, while 2 shows poor activity. Both 1 and 2 bind to DNA via groove‐binding. These results proved that the α‐hydroxy‐β‐oxo‐α,β‐enone moiety contributes to the pro‐oxidative activity of quercetin.     

Phylogeny of γ‐proteobacteria: resolution of one branch of the universal tree?

The reconstruction of bacterial evolutionary relationships has proven to be a daunting task because variable mutation rates and horizontal gene transfer (HGT) among species can cause grave incongruities between phylogenetic trees based on single genes. Recently, a highly robust phylogenetic tree was constructed for 13 gamma-proteobacteria using the combined alignments of 205 conserved orthologous proteins.1 Only two proteins had incongruent tree topologies, which were attributed to HGT between Pseudomonas species and Vibrio cholerae or enterics. While the evolutionary relationships among these species appears to be resolved, further analysis suggests that HGT events with other bacterial partners likely occurred; this alters the implicit assumption of gamma-proteobacteria monophyly. Thus, any thorough reconstruction of bacterial evolution must not only choose a suitable set of molecular markers but also strive to reduce potential bias in the selection of species.     

Mammals,agri‐environment schemes and set‐aside – what are the putative benefits?

• 1 The impacts of agricultural intensification on farmland wildlife have been the subject of increasing concern, particularly over the last two decades. Population declines have occurred for a number of mammalian species, sometimes drastically so, and changes in farming practice are believed to be significant contributory factors.
• 2 The major policy instruments for delivering environmental benefits on farmland are agri‐environment schemes. These encourage farmers to adopt more environmentally sensitive farming practices to promote farmland biodiversity. Additionally, compulsory set‐aside, which reduces agricultural surplus, could also have positive impacts on wildlife. In this paper we consider some of the putative benefits of agri‐environment schemes and set‐aside for mammals.
• 3 We review how establishment and management options within agri‐environment schemes and set‐aside might affect habitat resources for mammals. For example, conservation headlands increase plant and invertebrate resources within the crop edge for mammals such as wood mice. Grassy field margins can support communities of smaller mammals, and hedgerows may act as important commuting and hunting routes. Their potential will depend on factors such as seed mixtures used, timing and severity of cutting, and length of time they have been in place.
• 4 At a farm level, habitat heterogeneity may be increased through organic agriculture, which is supported by some agri‐environment schemes. Studies suggest significant benefits to mammals, including wood mice and bats. However, it is increasingly recognized that effective conservation of farmland mammals must seek solutions at the landscape scale, addressing such issues as habitat connectivity between farms. One approach may be the better targeting of scheme agreements.
• 5 We suggest that agri‐environment schemes and set‐aside can contribute to the conservation of mammals on farmland. Recent policy changes are likely to have further positive impacts on farmland wildlife but appropriate mammal monitoring programmes must be developed rigorously to assess their effects.

     

How Many Calories Are on Our Plate? Expected Fullness,Not Liking,Determines Meal‐size Selection

The availability of highly palatable food is thought to stimulate the selection of larger meals (leading to weight gain and obesity). In this article, we explore aspects of this proposition. Specifically, we scrutinize two basic assumptions: (i) palatable energy‐dense foods are more rewarding (desired), and (ii) these palatable foods are selected in relatively larger portions. In combination with palatability, we also consider the relative role for “expected satiation”—the extent to which a food is expected to deliver satiation. A total of 17 commonly consumed foods were assessed by 28 normal‐weight participants at lunchtime. Critically, our measure of food reward and expected satiation involves comparisons between foods based on equicaloric portions. When assessed in this way, we find that food reward and ideal portion sizes (in kcal) are both closely associated with expected satiation, but not with “expected liking.” Low expected satiation (not expected liking) predicts the selection of large portion sizes (in kcal) and foods with this characteristic tend to be more energy dense and are regarded as less (not more) rewarding (when compared calorie for calorie). Together, these findings challenge the role of palatability in meal‐size selection and they highlight the importance of expected satiation, a “nonaffective” component of food reward.     

How Many Seed Transfer Zones Are Necessary for the Preservation of the Genetic Identity of Austrocedrus chilensis Natural Populations in Argentina?

Two different studies based on isozymes that include genetic structure analysis have arrived at contrasting conclusions regarding the minimum number of seed transfer zones for Patagonian cypress (Austrocedrus chilensis) in Argentina that are required in order to avoid genetic contamination in restoration programs. Unfortunately, the more recent article lacks discussion on these controversial results, which is, therefore, the purpose of this article. The reliability of the markers used and the sampling performed in these studies are evaluated comparatively. The later study found higher levels of diversity and differentiation but paradoxically suggested that only two seed transfer zones would be enough to preserve the genetic identity of the natural populations of the species, whereas the earlier study concluded that at least five are necessary. Arguments are presented here for the case that definition of fewer than five genetically homogeneous groups is absolutely inappropriate and implies a probable risk of genetic contamination and maladaptation.     

Overproduction in the Lesser Black‐backed Gull – can marginal chicks overcome the initial handicap of hatching asynchrony?

In response to unpredictability of both food availability and core offspring failure, parents of many avian species initially produce more offspring than they commonly rear (overproduction). When parental investment is insufficient to raise the whole brood the handicap of hatching last means ‘marginal’ chicks are less likely to survive if brood reduction occurs. Conversely, if marginal offspring are required as replacements for failed ‘core’ chicks, or parental investment is sufficient to rear the whole brood, the handicap imposed on marginal chicks must be reversible if overproduction is to be a viable strategy. I investigated the ability of marginal offspring to overcome the handicap imposed by hatching asynchrony using a combination of a field experiment, designed to manipulate both the amount of total competition and the relative competitive ability of chicks within a brood, and data on the growth and survival of unmanipulated, three‐chick broods from three consecutive years. The results indicate that, even when resources are abundant, marginal offspring do not begin to overcome the competitive handicap imposed by hatching asynchrony until the period of growth when energetic requirements reach their peak, and subsequent survival to fledging is almost assured. This is apparently a consequence of parents controlling allocation of early parental investment, so that any brood reduction ‘decisions’ can be left as late as possible. Marginal chicks initially channel resources into maintaining mass, relative to skeletal size, as a buffer against starvation. However this also means competitiveness is reduced, so if conditions are poor marginal chicks are rapidly out‐competed, lose condition and die. Conversely, when food availability is good marginal offspring devote more resources to skeletal growth and quickly close the gap on their core siblings, meaning the handicap is reversible. The benefits of overproduction and hatching asynchrony as reproductive strategies to maximise success in Lesser Black‐backed Gulls are discussed in relation to the reproductive alternatives.     

Extraordinary Thermal Stability of an Oligodeoxynucleotide Octamer Constructed from Alternating 7‐Deaza‐7‐iodo guanine and 5‐Iodocytosine Base Pairs – DNA Duplex Stabilization by Halogen Bonds?

A reinvestigation of the published X‐ray crystal‐structure analyses of 7‐halogenated (Br, I) 8‐aza‐7‐deaza‐2′‐deoxyguanosines Br⁷c⁷z⁸G_d; 1a and I⁷c⁷z⁸G_d, 1b , as well as of the structurally related 7‐deaza‐7‐iodo‐2′‐deoxy‐β‐D ‐ribofuranosyladenine (β‐I⁷c⁷A_d; 2 = 6e in Table 1) and its α‐D ‐anomer (α‐I⁷c⁷A_d; 3 ) clearly revealed the existence of halogen bonds between corresponding halogen substituents and the adjacent N(3)‐atoms of neighboring nucleoside molecules within the single crystals. These halogen bonds can be rationalized by the presence of a region of positive electrostatic potential, the σ‐hole, on the outermost portion the halogen's surface, while the three unshared pairs of electrons produce a belt of negative electrostatic potential around the central part of the halogen substituent. The N(3) atoms of the halogenated nucleosides carry a partial negative charge. This novel type of bonding between nucleosides was tentatively used to explain the extraordinary high stability of oligodeoxynucleotides constructed from halogenated nucleotide building blocks.     

What is the role of the second “structural” NADP+‐binding site in human glucose 6‐phosphate dehydrogenase?

Human glucose 6-phosphate dehydrogenase, purified after overexpression in E. coli, was shown to contain one molecule/subunit of acid-extractable "structural" NADP+ and no NADPH. This tightly bound NADP+ was reduced by G6P, presumably following migration to the catalytic site. Gel-filtration yielded apoenzyme, devoid of bound NADP+ but, surprisingly, still fully active. Mr of the main component of "stripped" enzyme by gel filtration was approximately 100,000, suggesting a dimeric apoenzyme (subunit Mr = 59,000). Holoenzyme also contained tetramer molecules and, at high protein concentration, a dynamic equilibrium gave an apparent intermediate Mr of 150 kDa. Fluorescence titration of the stripped enzyme gave the K d for structural NADP+ as 37 nM, 200-fold lower than for "catalytic" NADP+. Structural NADP+ quenches 91% of protein fluorescence. At 37 degrees C, stripped enzyme, much less stable than holoenzyme, inactivated irreversibly within 2 d. Inactivation at 4 degrees C was partially reversed at room temperature, especially with added NADP+. Apoenzyme was immediately active, without any visible lag, in rapid-reaction studies. Human G6PD thus forms active dimer without structural NADP+. Apparently, the true role of the second, tightly bound NADP+ is to secure long-term stability. This fits the clinical pattern, G6PD deficiency affecting the long-lived non-nucleate erythrocyte. The Kd values for two class I mutants, G488S and G488V, were 273 nM and 480 nM, respectively (seven- and 13-fold elevated), matching the structural prediction of weakened structural NADP+ binding, which would explain decreased stability and consequent disease. Preparation of native apoenzyme and measurement of Kd constant for structural NADP+ will now allow quantitative assessment of this defect in clinical G6PD mutations.     

Are tyrosine residues involved in the photoconversion of the water‐soluble chlorophyll‐binding protein of Chenopodium album?

Non‐photosynthetic and hydrophilic chlorophyll (Chl) proteins, called water‐soluble Chl‐binding proteins (WSCPs), are distributed in various species of Chenopodiaceae, Amaranthaceae, Polygonaceae and Brassicaceae. Based on their photoconvertibility, WSCPs are categorised into two classes: Class I (photoconvertible) and Class II (non‐photoconvertible). Chenopodium album WSCP (CaWSCP; Class I) is able to convert the chlorin skeleton of Chl a into a bacteriochlorin‐like skeleton under light in the presence of molecular oxygen. Potassium iodide (KI) is a strong inhibitor of the photoconversion. Because KI attacks tyrosine residues in proteins, tyrosine residues in CaWSCP are considered to be important amino acid residues for the photoconversion. Recently, we identified the gene encoding CaWSCP and found that the mature region of CaWSCP contained four tyrosine residues: Tyr13, Tyr14, Tyr87 and Tyr134. To gain insight into the effect of the tyrosine residues on the photoconversion, we constructed 15 mutant proteins (Y13A, Y14A, Y87A, Y134A, Y13‐14A, Y13‐87A, Y13‐134A, Y14‐87A, Y14‐134A, Y87‐134A, Y13‐14‐87A, Y13‐14‐134A, Y13‐87‐134A, Y14‐87‐134A and Y13‐14‐87‐134A) using site‐directed mutagenesis. Amazingly, all the mutant proteins retained not only chlorophyll‐binding activity, but also photoconvertibility. Furthermore, we found that KI strongly inhibited the photoconversion of Y13‐14‐87‐134A. These findings indicated that the four tyrosine residues are not essential for the photoconversion.