Tuber-specific cytosolic expression of a bacterial phosphoglucomutase in potato (Solanum tuberosum L.) dramatically alters carbon partitioning

Constitutive antisense inhibition of the cytosolic isoform of phosphoglucomutase in the potato (Solanum tuberosum L.) results in restriction of photosynthesis, growth inhibition and modified tuber morphology, and a severe restriction of tuber starch synthesis. Here we describe the consequences of the tuber-specific expression of an Escherichia coli phosphoglucomutase in the cytosol. Analysis of [14C]glucose metabolism by tuber discs isolated from wild type and transformants revealed that the rates of sucrose and starch synthesis were unaltered but that the rate of glycolysis was depressed in the transgenics. The transformant tubers also contained dramatically reduced amino acid content and significantly higher levels of ADP, but were characterized by elevated levels of Krebs cycle intermediates and an unaltered rate of respiration. In addition to these metabolic consequences of the overexpression of the E. coli enzyme, we observed morphological changes in tubers, with the transformants having a smaller number of larger tubers which exhibited delayed rates of sprouting with respect to the wild type. These results are discussed with respect to current models of the regulation of central plant metabolism and tuber dormancy.     

Rhamnogalacturonan II structure shows variation in the side chains monosaccharide composition and methylation status within and across different plant species

A paradigm regarding rhamnogalacturonans II (RGII) is their strictly conserved structure within a given plant. We developed and employed a fast structural characterization method based on chromatography and mass spectrometry, allowing analysis of RGII side chains from microgram amounts of cell wall. We found that RGII structures are much more diverse than so far described. In chain A of wild‐type plants, up to 45% of the l –fucose is substituted by l –galactose, a state that is seemingly uncorrelated with RGII dimerization capacity. This led us to completely reinvestigate RGII structures of the Arabidopsis thaliana fucose‐deficient mutant mur1, which provided insights into RGII chain A biosynthesis, and suggested that chain A truncation, rather than l –fucose to l –galactose substitution, is responsible for the mur1 dwarf phenotype. Mass spectrometry data for chain A coupled with NMR analysis revealed a high degree of methyl esterification of its glucuronic acid, providing a plausible explanation for the puzzling RGII antibody recognition. The β–galacturonic acid of chain A exhibits up to two methyl etherifications in an organ‐specific manner. Combined with variation in the length of side chain B, this gives rise to a family of RGII structures instead of the unique structure described up to now. These findings pave the way for studies on the physiological roles of modulation of RGII composition.     

Modulation of intracellular Ca2+ release and capacitative Ca2+ entry by CaMKII inhibitors in bovine vascular endothelial cells

The effects of inhibitors of CaMKII on intracellular Ca²⁺ signaling were examined in single calf pulmonary artery endothelial (CPAE) cells using indo-1 microfluorometry to measure cytoplasmic Ca²⁺ concentration ([Ca²⁺]_i). The three CaMKII inhibitors, KN-93, KN-62, and autocamtide-2-related inhibitory peptide (AIP), all reduced the plateau phase of the [Ca²⁺]_i transient evoked by stimulation with extracellular ATP. Exposure to KN-93 or AIP alone in the presence of 2 mM extracellular Ca²⁺ resulted in a dose-dependent increase of [Ca²⁺]_i consisting of a rapid and transient Ca²⁺ spike followed by a small sustained plateau phase of elevated [Ca²⁺]_i. Exposure to KN-93 in the absence of extracellular Ca²⁺ caused a transient rise of [Ca²⁺]_i, suggesting that exposure to CaMKII inhibitors directly triggered release of Ca²⁺ from intracellular endoplasmic reticulum (ER) Ca²⁺ stores. Repetitive stimulation with KN-93 and ATP, respectively, revealed that both components released Ca²⁺ largely from the same store. Pretreatment of CPAE cells with the membrane-permeable inositol 1,4,5-trisphosphate (IP₃) receptor blocker 2-aminoethoxydiphenyl borate caused a significant inhibition of the KN-93-induced Ca²⁺ response, suggesting that exposure to KN-93 affects Ca²⁺ release from an IP₃-sensitive store. Depletion of Ca²⁺ stores by exposure to ATP or to the ER Ca²⁺ pump inhibitor thapsigargin triggered robust capacitative Ca²⁺ entry (CCE) signals in CPAE cells that could be blocked effectively with KN-93. The data suggest that in CPAE cells, CaMKII modulates Ca²⁺ handling at different levels. The use of CaMKII inhibitors revealed that in CPAE cells, the most profound effects of CaMKII are inhibition of release of Ca²⁺ from intracellular stores and activation of CCE. Ca²⁺/calmodulin-dependent kinase II; calcium regulation; capacitative calcium entry     

Competitive Mirror Image Phage Display Derived Peptide Modulates Amyloid Beta Aggregation and Toxicity

Alzheimer´s disease is the most prominent type of dementia and currently no causative treatment is available. According to recent studies, oligomeric species of the amyloid beta (Aβ) peptide appear to be the most toxic Aβ assemblies. Aβ monomers, however, may be not toxic per se and may even have a neuroprotective role. Here we describe a competitive mirror image phage display procedure that allowed us to identify preferentially Aβ_1–42 monomer binding and thereby stabilizing peptides, which destabilize and thereby eliminate toxic oligomer species. One of the peptides, called Mosd1 (monomer specific d-peptide 1), was characterized in more detail. Mosd1 abolished oligomers from a mixture of Aβ_1–42 species, reduced Aβ_1–42 toxicity in cell culture, and restored the physiological phenotype in neuronal cells stably transfected with the gene coding for human amyloid precursor protein.     

Protoprimulagenin a Als Aglykon des Hauptsaponins von Primula elatior

It was shown that the aglycone of the main saponin of Primula elatior (L.) Hill is not primulagenin A but protoprimulagenin A with an oxide ring between C-13 β and C-28 in the β amyrin ring system.     

1,4,9,10-Anthradiquinone as precursor for antitumor compounds

1,4,9,10-Anthradiquinone 5 was reacted with enamines 6 in the Nenitzescu reaction to yield unexpected 3,3a,6,12-tetrahydro-3a,7-dihydroxy-2-methyl-6,12-dioxo-naphtho[2,3-d]indol-1-carboxylates 8A. However, anthracycline-like naphtho-condensed 5-hydroxyindoles were not obtained from this diquinone. It yielded similar reaction products of the Nenitzescu reaction like other quinones activated by two electron-withdrawing groups. Furthermore, these new compounds 8A were found to constitute precursors for the synthesis of azonines. The conversion to dibenzoazonines 13 occurred in an unusual and up to now unknown way consisting of isomerization, ring opening, and re-closure. 2-Chloro-anthradiquinone 19 reacted with enamines 6 as vinylogeous acid chloride to pyrroloanthraquinone 20. No substitution of chlorine was observed. Naphtho-condensed indoles 26 were obtained by the reactions of unsubstituted 1,4-anthraquinone 25 with enamines 6 via the normal Nenitzescu route. Indoles 26 were converted to Mannich bases, reacting further to dimers by the Diels-Alder reaction of intermediate o-quinone methides. Most of the synthesized heterocycles were evaluated for their anticancer properties in the NCI's human-disease oriented in vitro anticancer screen. Particularly, carbinolamines 8A exhibited inhibitory activity of tumor cell growth and thus they constitute a new class of lead structures for anticancer drug design.     

Über eine postmeiotische Teilungsanomalie und den Spiralbau der Chromosomen von Paris quadrifolia

Zusammenfassung In drei Pflanzen einer Kolonie von Paris quadrifolia wurde in eben entstandenen Gonen eine abnorme postmeiotische Mitose beobachtet, die bis zur Metaphase geht und dann rückläufig über eine Telophase zu einem meist normalen Ruhekern führt. Die Chromosomen sind ungespalten und entsprechen äußerlich und innerlich den Anaphasechromosomen der homöotypischen Teilung. Obwohl diese Chromosomen die Wertigkeit von Chromatiden besitzen, also keine teilungsfähigen, aus zwei Chromatiden aufgebauten Vollchromosomen sind, erfolgt die Spindelbildung, die Metakinesebewegung und die Orientierung der Chromosomen in der Äquatorialplatte normal. Diese Vorgänge sind also unabhängig vom Alter der Chromosomen und Centromeren. Auch die Einstellung der Spindel in der Zelle unter Drehung des Polfeldes erfolgt so, wie es zu erwarten wäre, wenn eine normale Zellteilung an dieser Stelle stattfände.Die Spindeleinstellung der abnormen Mitose ist mechanisch, bedingt und eine andere als in der 1. Pollenmitose, bei der nicht einfach mechanische Gesetzmäßigkeiten wirken, sondern eine bestimmte plasmatische Differenzierung bestimmend ist.Das Auftreten der postmeiotischen Mitose zeigt keine ursächliche Beziehung zu den für Paris bezeichnenden Störungen infolge von Inversionsheterozygotie. Die Ursache kann genotypischer oder phänotypischer Natur sein; für beide Annahmen lassen sich Anhaltspunkte gewinnen.Durch Vorbehandlung mit NH³-AIkohol läßt sich der Spiralbau der Chromosomen in der 1. Pollenmitose klar sichtbar machen. Es bestätigt sich die Auffassung, daß je Chromatide eine Doppelspirale vorhanden ist, daß aber nicht zwei auseinandergeschobene Spiralen vorliegen. Die Großspiralisierung kann als Modell dienen um Deutungen vorzunehmen, wo die unmittelbare optische Beobachtung versagt.     

Arginine deiminase inhibits cell proliferation by arresting cell cycle and inducing apoptosis.

We have previously demonstrated that arginine deiminase inhibits the proliferation of vascular endothelial cells, but the mechanisms leading to growth inhibition have remained unclear. We report here that low concentrations of arginine deiminase purified from Mycoplasma arginini inhibit proliferation of various cultured cells by arresting the cell cycle in G(1) and/or S phase with higher arginine deiminase concentrations leading to subsequent apoptosis. Our results demonstrate that arginine deiminase inhibits cell proliferation not only by depletion of arginine, but also by mechanisms involving the cell cycle and death signals.     

Characterization of attaching and effacing <Emphasis Type="Italic">Escherichia coli</Emphasis> (AEEC) isolated from pigs and sheep

Background  

Attaching and effacing Escherichia coli (AEEC) are characterized by their ability to cause attaching-and-effacing (A/E) lesions in the gut mucosa of human and animal hosts leading to diarrhoea. The genetic determinants for the production of A/E lesions are located on the locus of enterocyte effacement (LEE), a pathogenicity island that also contains the genes encoding intimin (eae). This study reports data on the occurrence of eae positive E. coli carried by healthy pigs and sheep at the point of slaughter, and on serotypes, intimin variants, and further virulence factors of isolated AEEC strains.