Combinatorial synthesis of peptide arrays onto a microchip

Arrays promise to advance biology through parallel screening for binding partners. We show the combinatorial in situ synthesis of 40,000 peptide spots per square centimeter on a microchip. Our variant Merrifield synthesis immobilizes activated amino acids as monomers within particles, which are successively attracted by electric fields generated on each pixel electrode of the chip. With all different amino acids addressed, particles are melted at once to initiate coupling. Repetitive coupling cycles should allow for the translation of whole proteomes into arrays of overlapping peptides that could be used for proteome research and antibody profiling.     

Video-rate far-field optical nanoscopy dissects synaptic vesicle movement

We present video-rate (28 frames per second) far-field optical imaging with a focal spot size of 62 nanometers in living cells. Fluorescently labeled synaptic vesicles inside the axons of cultured neurons were recorded with stimulated emission depletion (STED) microscopy in a 2.5-micrometer by 1.8-micrometer field of view. By reducing the cross-sectional area of the focal spot by about a factor of 18 below the diffraction limit (260 nanometers), STED allowed us to map and describe the vesicle mobility within the highly confined space of synaptic boutons. Although restricted within boutons, the vesicle movement was substantially faster in nonbouton areas, consistent with the observation that a sizable vesicle pool continuously transits through the axons. Our study demonstrates the emerging ability of optical microscopy to investigate intracellular physiological processes on the nanoscale in real time.     

Effects of increasing dietary rye levels on physicochemical characteristics of digesta and its impact on stomach emptying as well as the formation of ‘doughballs’ in stomachs of young pigs

Despite quite similar contents of starch and crude fibre of wheat and rye, the unique non-starch-polysaccharide fraction of rye (e.g. high levels of arabinoxylans and fructans) might have an impact on physicochemical properties of the digesta in pigs. Forty pigs (age: 46.8 ± 5.28 days; bodyweight: 16.1 ± 4.13 kg) were divided into four treatment groups. During four weeks, the pigs received diets consisting of wheat and/or rye, barley, soy, potato protein and a mineral supplement. The sum of wheat and rye was 69% in all diets, whereby the compound feed of each group was characterized by a different ratio (%) of wheat/rye (69/0; 46/23; 23/46; 0/69, respectively). In the stomach, ‘doughballs’ occurred more frequently with increasing dietary rye levels (9/10; 69% rye). With higher DM content and extract-viscosity of gastric digesta, the stomach emptying tended to be retarded in rye groups. Compared to the control group (69% wheat), maximum dietary rye levels (69%) resulted in significantly higher concentrations of lactic acid in digesta of the stomach and small intestine. With increasing lactic acid concentrations, the pH tended to be lower in small intestinal digesta. With an intensified formation of lactic acid, effects against Gram-negative bacteria, for example Salmonella, can be expected. Moreover, because of higher viscosity and the retarded stomach emptying, there could be advantages of including rye in compound feeds when a longer lasting satiety is intended, for example when feeding pregnant sows (regularly fed restrictively).     

Überprüfung eines Schätzverfahrens zur Ermittlung von Kennwerten der Wasserbindung anhand der Labordatenbank des Niedersächsischen Bodeninformationssystems

Testing a method for estimating water retention parameters using the laboratory database of the Lower Saxony Soil Information System The validity of the method used for estimating field capacity (pF > 1.8), plant available water (pF 1.8 – 4.2), air capacity (pF < 1.8), and total pore volume from soil texture, packing density (bulk density + 0.009 % clay) and humus content described by the Arbeitsgruppe Bodenkunde (1982) was checked on the basis of 1693 pF curves of the laboratory database of the Lower Saxony Soil Information System (NIBIS). The positive and negative corrections for humus content applied in this method to the above parameters are clearly too small. Use of tables for estimating the pore volume of humus-free soils leads to overestimation. It will only be possible to work out an alternative method applicable to all classes of soils when the database has been extended.     

Ermittlung bodenphysikalischer Kennwerte in Abhängigkeit von Bodenart,Lagerungsdichte und Humusgehalt

Evaluation of soil physical parameters as a function of soil texture, effective density and humus content The latest, 4. edition of the Soil Mapping Instruction presents a new soil texture diagram with 31 partially newly defined soil textures. This diagram works without alternative names of clay silt mixtures, it takes into account the latest scientific findings and should obligatory be used in the new and old Bundesländer of the BRD. Though the soil textures define almost all soil specific valuations, the soil physical parameters depending from texture had to be recalculated on an enlarged database. 6352 records from 8 Bundesländer were used on this calculation. Out of these records most of the 31 soil textures could be characterized by representative and statistically proved values of air capacity, available water capacity, field moisture capacity and mean water permeability as a function of bulk density, dry, effective density and humus content. The values are presented in tables.     

Bemessung von Meßintervallen für Dauerbeobachtungsflächen in Boden-Meßnetzen

Calculation of time intervals for soil analysis in continuous soil measuring units In continuous soil measuring units in the field the intervals between soil analysis which differentiate data in a statistical reliable manner are unknown. A method is proposed to calculate the above mentioned intervals and an example is calculated. For a Parabraunerde from loess under decidious forest in Baden-Württemberg with low immission of Pb the intervals between soil analysis should be 40 years.     

Alterations of vitamin C, total phenolics, and antioxidant capacity as affected by processing tomatoes to different products

This study was conducted to investigate the antioxidant vitamin C, the polyphenol content, and the hydrophilic antioxidant capacity of tomato juice, baked tomatoes, tomato sauce, and tomato soup. During the production of tomato juice and during the preparation of the other tomato products, samples were taken after different times, respectively, after each particular production step. High-performance liquid chromatography was used to determine the content of vitamin C. The total phenolics content was analyzed spectrophotometrically by using the Folin-Ciocalteu method. The hydrophilic antioxidant capacity was measured by using three different methods: the Trolox equivalent antioxidant capacity assay, the ferric reducing antioxidant power test, and the photochemiluminescence assay. The vitamin C contents of the tomato products decreased during the thermal processing of tomatoes. In contrast, the total phenolics concentration and the water soluble antioxidant capacity increased.     

Uptake and apoplastic retention of EDTA‐ and phytosiderophore‐chelated chromium(III) in maize

Increasing the mobilization and root uptake of chromium (Cr) by synthetic and plant‐borne chelators might be relevant for the design of phytoremediation strategies on Cr‐contaminated sites. Short‐term uptake studies in maize roots supplied with ⁵¹CrCl₃ or ⁵¹Cr(III)‐EDTA led to higher apoplastic Cr contents in plant roots supplied with ⁵¹CrCl₃ and in Fe‐sufficient plants relative to Fe‐deficient plants, indicating that Fe stimulated co‐precipitation of Cr. Concentration‐dependent retention of Cr in a methanol:chloroform‐treated cell‐wall fraction was still saturable and in agreement with the predicted tendency of Cr(III) to precipitate as Cr(OH)₃. To investigate a possible stimulation of Cr(III) uptake by phytosiderophores, Fe‐deficient maize roots were exposed for 6 d to Cr(III)‐EDTA or Cr(III)‐DMA (2'‐deoxymugineic acid). Relative to plants without Cr supply, the supply of both chelated Cr species in a subtoxic concentration of 1 µM resulted in alleviation of Fe deficiency–induced chlorosis and higher Cr accumulation. Long‐term Cr accumulation from Cr(III)‐DMA was similar to that from Cr(III)‐EDTA, and Cr uptake from both chelates was not altered in the maize mutant ys1, which is defective in metal‐phytosiderophore uptake. We therefore conclude that phytosiderophores increase Cr solubility similar to synthetic chelators like EDTA, but do not additionally contribute to Cr(III) uptake from Cr‐contaminated sites.     

Nonresolvable Row–Column Designs with an Even Distribution of Treatment Replications

When generating experimental designs for field trials laid out on a rectangular grid of plots, it is useful to allow for blocking in both rows and columns. When the design is nonresolvable, randomized classical row–column designs may occasionally involve clustered placement of several replications of a treatment. In our experience, this feature prevents the more frequent use of these useful designs in practice. Practitioners often prefer a more even distribution of treatment replications. In this paper we illustrate how spatial variance–covariance structures can be used to achieve a more even distribution of treatment replications across the field and how such designs compare with classical row–column designs in terms of efficiency factors. We consider both equally and unequally replicated designs, including partially replicated designs. Supplementary materials accompanying this paper appear online.     

Physiological and molecular characterization of Fe acquisition by tomato plants from natural Fe complexes

The aim of this work is to evaluate the capability of tomato plants to use different Fe sources, such as Fe citrate, Fe phytosiderophores, and Fe complexed by a water-extractable humic substances (Fe-WEHS) also in relation to physiological and molecular adaptations induced by these complexes at the root level. Tomato plants acquired higher amounts of Fe from Fe-WEHS than from the other two sources and this phenomenon occurred only when the treatment lasted 24 h. The higher acquisition of Fe from Fe-WEHS than other sources depended on a reductive mechanism and on rhizosphere acidification and appeared to be due neither to a higher apoplastic loading nor to a higher resistance of WEHS to microbial degradation. Supply of the different Fe complexes to deficient plants induced a transient upregulation of Fe(III)-chelate reductase (LeFRO1) and Fe transporter genes, LeIRT1 and LeIRT2. In Fe-WEHS-fed plants, where a quicker and higher upregulation of these genes was evident, a coordination in the expression of LeFRO1, LeIRT1, and LeIRT2 genes occurred already after 1 h treatment when the amount of Fe acquired by the plants from the three sources was similar. Iron from Fe-WEHS could be efficiently acquired in a mixture of natural Fe complexes possibly occurring in the rhizosphere. This phenomenon is due to an altered expression of Fe uptake-related genes and to the root capacity to create favorable conditions for the micronutrient uptake into the rhizosphere.