Estimation of plasmalemma conductivity to ascorbic acid in intact leaves exposed to ozone

To establish the capacity of the leaf mesophyll plasmalemma of Phaseolus vulgaris L. to supply ascorbate (ASC) into the cell wall by simple diffusion, a method for calculating plasmalemma diffusional conductivity to ascorbic acid (AA) in intact leaves was evaluated. The core of the approach is that in the presence of a sink for ascorbate in the cell wall, cell wall total ascorbic acid concentration [TAA]_cw (=[ASC]_cw+[AA]_cw) reaches zero at some positive whole‐leaf total ascorbic acid concentration [TAA]_l. It is shown that [TAA]_l at [TAA]_cw=0 is proportional to the sink for ASC in the cell wall and the reciprocal of plasmalemma conductivity. The predicted proportional relationship between [TAA]_cw and [TAA]_l was confirmed by decreasing TAA levels in leaves through predarkening. Furthermore, increasing the sink intensity for ASC in the cell wall by the acute exposure of leaves to 450 nmol ozone mol^?1 during re‐illumination, [TAA]_cw reached zero at 2.7‐fold higher [TAA]_l than without ozone, and the slope of the relationship increased twofold. Plasmalemma diffusional conductivities to AA of 2.9×10^?6 and 1.8×10^?6 m s^?1, needed to maintain [TAA]_cw at the observed level, were calculated from the increase in [TAA]_l at [TAA]_cw=0 and from the two different estimates of the sink for ASC. A value of 1.3×10^?6 m s^?1 was calculated on the basis of the oil‐water distribution coefficient for TAA. It is concluded that the demand for ASC in the mesophyll cell wall of the investigated leaves could be met by simple diffusion of AA through the plasmalemma. From the measured increase in the slope of the relationship [TAA]_cw versus [TAA]_l, an increase in the cell wall pH of 0.3 units was estimated under the influence of ozone.     

Copper(II) binding to the human Doppel protein may mark its functional diversity from the prion protein

Doppel (Dpl) is the first described homologue of the prion protein, the main constituent of the agent responsible for prion diseases. The cellular prion protein (PrP(C)) is predominantly present in the central nervous system. Although its role is not yet completely clarified, PrP(C) seems to be involved in Cu(2+) recycling from synaptic clefts and in preventing neuronal oxidative damage. Conversely, Dpl is expressed in heart and testis and has been shown to regulate male fertility by intervening in gametogenesis and sperm-egg interactions. Therefore, despite a high sequence homology and a similar three-dimensional fold, the functions of PrP(C) and Dpl appear unrelated. Here we show by electron paramagnetic resonance and fluorescence spectroscopy that the in vitro binding of copper(II) to human recombinant Dpl occurs with a different pattern from that observed for recombinant PrP. At physiological pH values, two copper(II)-binding sites with different affinities were found in Dpl. At lower pH values, two additional copper(II)-binding sites can be identified as follows: one complex is present only at pH 4, and the other is observed in the pH range 5-6. As derived from the electron paramagnetic resonance characteristics, all Dpl-copper(II) complexes have a different coordination sphere from those present in PrP. Furthermore, in contrast to the effect shown previously for PrP(C), addition of Cu(2+) to Dpl-expressing cells does not cause Dpl internalization. These results suggest that binding of the ion to PrP(C) and Dpl may contribute to the different functional roles ascribed to these highly homologous proteins.     

A simple method of DNA extraction from soil for detection of composite transgenic plants by PCR

This new and simple method of DNA extraction from composite soil allows the isolation of plant DNA with high efficiency, quality and reproductivity. The method is based on a simple CaCl₂-precipitation step and requires no additional purification steps to eliminate humic acids. The extracted DNA was obtained in sufficient purity and quantity to allow direct detection of transgenes by PCR. Furthermore, the simple procedure allows the assay of many samples at the same time.     

Glycopeptide biosynthesis in the context of basic cellular functions

Using molecular genetics, biochemistry and organic chemistry the biosynthesis of glycopeptides has been elucidated in detail. It can be categorised in three parts: precursor supply, linking of the peptide backbone and modification reactions. The important steps of the biosynthesis are carried out at a multi-enzyme complex consisting of three non-ribosomal peptide synthetases (NRPS), three oxygenases and one halogenase. Novel derivatives can be generated by precursor-directed biosynthesis or combinatorial approaches and the knowledge can be used to optimise the yield of production by metabolic engineering approaches. To protect themselves glycopeptide producers seem to have developed strategies which may differ from those of the resistant pathogens.     

Synthetic biology of secondary metabolite biosynthesis in actinomycetes: Engineering precursor supply as a way to optimize antibiotic production

Actinomycetes are a rich source for the synthesis of medically and technically useful natural products. The genes encoding the enzymes for their biosynthesis are normally organized in gene clusters, which include also the information for resistance (in the case of antibacterial compounds), regulation, and transport. This facilitates the manipulation of such pathways by molecular genetic techniques. Recent advances in DNA sequencing and analytical chemistry revealed that not only new strains isolated from yet unexplored habitats, but also already known strains possess a large potential for the synthesis of novel compounds. Synthetic Biology now offers a new perspective to exploit this potential further by generating novel pathways, and thereby novel products, by combining different biosynthetic steps originating from different bacteria. The supply of precursors, which are subsequently incorporated into the final product, is often already organized in a modular manner in nature and may directly be exploited for Synthetic Biology. Here we report examples for the synthesis of building blocks and possibilities to modify and optimize antibiotic biosynthesis, exemplary for the synthesis of the manipulation of the synthesis of the glycopeptide antibiotic balhimycin.     

Testing a postulated case of intersexual selection in humans: The role of foot size in judgments of physical attractiveness and age

The constituents of attractiveness differ across the sexes. Many relevant traits are dimorphic, suggesting that they are the product of intersexual selection. However, direction of causality is generally difficult to determine, as aesthetic criteria can as readily result from, as cause, dimorphism. Women have proportionately smaller feet than men. Prior work on the role of foot size in attractiveness suggests an asymmetry across the sexes, as small feet enhance female appearance, yet average, rather than large, feet are preferred on men. Previous investigations employed crude stimuli and limited samples. Here, we report on multiple cross-cultural studies designed to overcome these limitations. With the exception of one rural society, we find that small foot size is preferred when judging women, yet no equivalent preference applies to men. Similarly, consonant with the thesis that a preference for youth underlies intersexual selection acting on women, we document an inverse relationship between foot size and perceived age. Examination of preferences regarding, and inferences from, feet viewed in isolation suggests different roles for proportionality and absolute size in judgments of female and male bodies. Although the majority of these results bolster the conclusion that pedal dimorphism is the product of intersexual selection, the picture is complicated by the reversal of the usual preference for small female feet found in one rural society. While possibly explicable in terms of greater emphasis on female economic productivity relative to beauty, the latter finding underscores the importance of employing diverse samples when exploring postulated evolved aesthetic preferences.