Targeted On‐line SPE‐LC‐MS/MS Assay for the Quantitation of 12 Apolipoproteins from Human Blood

Laborious sample pretreatment of biological samples represents the most limiting factor for the translation of targeted proteomics assays from research to clinical routine. An optimized method for the simultaneous quantitation of 12 major apolipoproteins (apos) combining on‐line SPE and fast LC‐MS/MS analysis in 6.5 min total run time was developed, reducing the manual sample pretreatment time of 3 μL serum or plasma by 60%. Within‐run and between‐day imprecisions below 10 and 15% (n = 10) and high recovery rates (94–131%) were obtained applying the high‐throughput setup. High‐quality porcine trypsin was used, which outperformed cost‐effective bovine trypsin regarding digestion efficiency. Comparisons with immunoassays and another LC‐MS/MS assay demonstrated good correlation (Pearson's R: 0.81–0.98). Further, requirements on sample quality concerning sampling, processing, and long‐term storage up to 1 year were investigated revealing significant influences of the applied sampling material and coagulant on quantitation results. Apo profiles of 1339 subjects of the LIFE‐Adult‐Study were associated with lifestyle and physiological parameters as well as establish parameters of lipid metabolism (e.g., triglycerides, cholesterol). Besides gender effects, most significant impact was seen regarding lipid‐lowering medication. In conclusion, this novel highly standardized, high‐throughput targeted proteomics assay utilizes a fast, simultaneous analysis of 12 apos from least sample amounts.     

Density-dependent shift from facilitation to competition in a dwarf <Emphasis Type="Italic">Avicennia germinans</Emphasis> forest

The global effort to rehabilitate and restore destroyed mangrove forests is unable to keep up with the high mangrove deforestation rates, which exceed the average pace of global deforestation. Although facilitation theory presents new possibilities for the restoration of heavily degraded mangrove sites, knowledge of tree–tree interactions in stressed mangrove forest ecosystems is too limited to utilize facilitation appropriately. The aim was to determine the mode of local interaction among stressed mangrove trees by investigating the effect of clustering on tree size and crown morphology under contrasting stand densities. The study was conducted in a dwarf Avicennia germinans forest in Northern Brazil, in which tree growth is limited by infrequent inundation and high pore-water salinity. Autoregressive regression, Voronoi tessellation and spatial point pattern statistics were used to address the spatial processes underlying tree interaction. Under low stand density (1.2 trees m^?2) dwarf trees which grew in clustered cohorts of A. germinans had a less stunted crown morphology revealing the dominance of a positive neighborhood influence among plants. In contrast, dwarf trees in the denser forest stand (2.7 trees m^?2) were interacting competitively as indicated by the more negative effect of neighbors on crown morphology and size. The shift from facilitative to competitive interactions is an important feature of mangrove forest regeneration under harsh environmental conditions. If mangrove trees are unable to regenerate naturally on severely degraded sites, intraspecific facilitation could be used to assist regeneration by planting seedlings in clusters and not evenly spaced.     

Identification of a Gene (GPR30) with Homology to the G-Protein-Coupled Receptor Superfamily Associated with Estrogen Receptor Expression in Breast Cancer

Using the technique of differential cDNA library screening, a cDNA clone was isolated from an estrogen receptor (ER)-positive breast carcinoma cell line (MCF7) cDNA library based upon the overexpression of this gene compared to an ER-negative cell line (MDA-MB-231). Sequence analysis of this clone determined that it shared significant homology to G-protein-coupled receptors. This receptor, GPCR-Br, was abundantly expressed in the ER-positive breast carcinoma cell lines MCF7, T-47D, and MDA-MB-361. Expression was absent or minimal in the ER-negative breast carcinoma cell lines BT-20, MDA-MB-231, and HBL-100. GPCR-Br was ubiquitously expressed in human tissues examined but was most abundant in placenta. GPCR-Br expression was examined in 11 primary breast carcinomas. GPCR-Br was detected in all 4 ER-positive tumors and only 1 of 7 ER-negative tumors. Based upon PCR analysis in hybrid cell lines, the gene for GPCR-Br (HGMW-approved symbol GPR30) was mapped to chromosome 7p22. The pattern of expression of GPCR-Br indicates that this receptor may be involved in physiologic responses specific to hormonally responsive tissues.     

Reproductive Behaviour in Zonocerus elegans (Orthoptera: Pyrgomorphidae) with Special Reference to Nuptial Gift Guarding

Egglaying aggregations, mate fidelity and male and female mate choice and mating behaviour of the African pyrgomorphid grasshopper Zonocerus elegans are described from the field. Several hundred males and females were individually marked at oviposition sites. Pairs remained stable over days, during egglaying and presumably also over the weeks prior to egglaying. In addition to long-term consorting with a female, males may try to obtain inseminations by “footpad”–tactics at oviposition sites. It is suggested that prolonged consorting and repeated copulations increase the female's reproductive output as well as the male's share in it. Our data and the literature indicate that males are involved not only in rival competition but also in a “battle of the sexes” which females are likely to win. The intra– and intersexual “war on two fronts” results behaviourally in elaborate mating tactics and anatomically in highly developed internal female genitalia and male copulatory organs (well known taxonomic characters). This applies not only to grasshoppers but also to other insects and arthropods.     

Localization of beta-glycerophosphatase and Mg++-activated adenosine triphosphatase in a moss haustorium,and the relation of these enzymes to the cell wall labyrinth

Summary Acid phosphatase and ATPase were localized in the bryophyte haustorium and in the surrounding paternal tissue of the gametophyte. Only cells with wall labyrinths are the sites of intense enzyme activity.The reaction products of both enzymes were found to occur in cell organelles, the plasma membrane and particularly in wall inclusions that are supposedly proteinaceous in nature.The intensity of the reaction is proportional to the state of differentiation of the labyrinth. The highest enzyme activity was encountered in the epidermal cells which are furnished with the most elaborate wall labyrinths. Somewhat lower was the activity in the other labyrinth cells with filiform ingrowths.These findings stress the role of the epidermis as an absorptive epithelium, and also show clearly that the enzyme activity associated with the plasma surface is in no reciprocal ratio to the degree of amplification of the surface area.     

Dorsiventralität der Statocyten in plagiogeotropen Seitenwurzeln von Lepidium sativum L.

Summary The calyptra of plagiotropic lateral roots of Lepidium sativum L. is composed of three rows of cells. Movable amyloplates, possibly functioning as statoliths, are located only a few central cells of the ontogenetic youngest cell row. Beside the lateral root axis the two innermost statocytes contain a stable complex of rough endoplasmic reticulum, which is preferentially located in the central distal cell corner. In the statocytes lying above the lateral root axis the amyloplasts are sedimented on the ER-complex during growth in direction of the geotropic liminal angle. In the statocyte below the axis the ER-complex is free of amyloplasts. Thus a dorsiventrality exists in the statocytes located above and below the root axis in regard to the arrangement of their organelles.

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Herrn Professor Dr. Maximilian Steiner zum 70. Geburtstag.     

Induction of multiple root systems in apple seedlings by radicle splitting

Summary A technique is described where germinating apple seeds are induced to form a divided root system. Plants with two or four evenly divided roots are produced by cutting the seed radicle before germination. The advantages of this method over the simple splitting of an existing root system are discussed.     

Die Struktur der Hoch Endopolyploiden Kerne Im Endosperm von Zea Mays,Das Auffallende Verhalten Ihrer Nucleolen und Ihr Endopolyploidiegrad

Zusammenfassung Die endopolyploiden Kerne des Endosperms von Zea mays enthalten bei 3 eingehend und 10 stichprobenartig untersuchten Sippen durchgehend Chromatinstränge, die sich aus den perlschnurartig gebauten Endochromosomen zusammensetzen. Sie stehen zum Teil netzartig miteinander in Verbindung, so daß sie nicht individuell als Riesenchromosomen kenntlich sind. In ihren Verlauf schalten sich die endomitotisch herangewachsenen knobs in Form kompakter Endochromozentren oder±dichter Ansammlungen von Heterochromomeren ein.Die Endopolyploidisierung führt bei den drei Sippen WF 9, M 14 und WF 9 X M 14 und offenbar auch bei den zehn anderen bis zu 192-Ploidie, ausnahmsweise zu 384-Ploidie.Die Kerne von M 14 erscheinen bei höchstwahrscheinlich gleicher Polyploidiestufe im allgemeinen ärmer an Chromatin als die von WF 9, und auch innerhalb dieser Sippe gibt es analoge Schwankungen kleineren Ausmaßes. Sie beruhen vermutlich auf Spiralisierungseffekten.Sippenspezifische Unterschiede bestehen auch im Vorkommen von Sekundärnucleolen, kleineren Nucleolen, die dicht neben den Hauptnucleolen an den drei endo-nucleolar-organizing bodies entstehen und bei WF 9 häufiger, in höherer Zahl und größeren Dimensionen als bei M 14 auftreten. Bei der Sippe M 14 werden dagegen häufiger und gehäuft Vacuolen aus den Nucleolen ausgestoßen (=freie N.); auch finden sich reichlicher als bei WF 9 zusätzliche kleine Nucleolen in den Chromatinsträngen eingeschaltet oder an ihnen angeheftet (=festsitzende N.). Der Bastard nimmt bezüglich der Sekundärnucleolen eine Mittelstellung zwischen den Elternsippen ein; bezüglich der freien und festsitzenden Nucleolen verhält er sich ähnlich wie M 14, bezüglich der Protuberanzen ähnlich wie WF 9.Die verschiedenartigen Nucleolen—einschließlich der in Ausstoßung begriffenen Nucleolusvacuolen—enthalten RNS, was sich aus der Methylgrün-Pyroninfärbung, kombiniert mit Ribonuclease-Behandlung ergibt.

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The structure of the highly endopolyploid nuclei in the endosperm of Zea mays, their conspicuous nucleolar conditions and degree of endopolyploidy

Summary The endopolypoid nuclei in the endosperm of Zea mays (8 inbred lines and 5 hybrids, 2+1 were investigated intensively) throughout contain strands of chromatin, which are composed of endochromosomes having chromomeric constitution. Some of the strands join laterally and most of them cannot be recognized individually. Inserted within them are endopolyploid knobs of compact heterochromatin or assemblages of heterochromomeres.In the lines WF 9 and M 14, the hybrid WF 9 x M 14 and obviously in the others the largest nuclei are 192 n and exceptionally 384 n.Nuclei of M 14 most likely having the same degree of polyploidy as those of WF 9 seem to contain less chromatin and even within WF 9 there are similar variations; probably they result from different spiralization.The different lines also show specific differences concerning the secondary nucleoli; these are nucleoli developing with endopolyploidization side by side with the main nucleoli at the three endo-nucleolar-organizing-bodies; WF 9 shows them in more nuclei, in higher frequency and in larger dimensions than M 14. In M 14 on the contrary more vacuoles (=free nucleoli) are ejected from the nucleoli and this happens more often; small additional nucleoli are also more abundantly inserted in the chromatin-strands. The hybrid behaves intermediately concerning the secondary nucleoli, more like M 14 concerning inserted and free nucleoli and similar to WF 9 in expulsion of vacuoles.Staining with methylgreen-pyronin and treatment with ribonuclease show the different nucleoli including the ejected vacuoles to contain RNA.