Die Haploidie der MÄnnchen und die Endopolyploidie in Einigen Geweben von Haplothrips (Thysanoptera)

Zusammenfassung Die Spermatogonien sind haploid, die Oögonien diploid, die Chromosomenzahl beträgt bei Haplothrips statices n=15. Die Ganglienzellen und die Nervenmutterzellen sind bei Männchen haploid, bei Weibchen diploid.Haploid sind bei den Männchen auch die Zellen der Epidermis, der Tracheenmatrix und des Hinterdarniepithels mindestens bis zur Pronymphe.Es findet demnach während der Entwicklung der von Haplothrips keine allgemeine Aufregulierung (Diploidisierung) der Zellen statt.Fettkörper, Mitteldarmepithel, Malpighigefäße und Oenocyten werden polyploid bis zu 32n. Dabei teilen sich im Fettkörper mindestens noch die 16-ploiden Zellkerne. Während im Mitteldarmepithel, den Malpighigefäßen und vermutlich auch im Fettkörper das Verhältnis der Polyploidie von l2 entsprechend der haploiden Ausgangsbasis der männlichen Zellen erhalten bleibt, wächst die Mehrzal der Oenocyten bei den Männchen stärker als bei den Weibchen.Mit Unterstützung der Deutschen Forschungsgemeinschaft.     

Die Giltigkeit des Energiemengengesetzes für den negativen Galvanotropismus der Wurzel

Ohne Zusammenfassung     

Simple emission-reducing measures in an open biological waste treatment plant

In the course of composting biological waste, concentrations of various thermophilic and thermotolerant microorganisms increase. Moving piles of compost results in increased emissions of Actinomycetes and fungi. The present investigation deals with the reduction of airborne microorganism emission and immission in large-scale composting plants with open piles. Simple measures were introduced in order to reduce the release of bioaerosols when turning piles and the release of dust and bioaerols at large. These measures included the sealing of turning machinery with rubber mats, the wetting of piles before and after turning and regular cleaning and wetting driveways during the dry season.Concentrations of airborne microorganisms during the summer season were determined on 5 days before and after the introduction of emission-reducing measures using the six-stage Andersen cascade sampler. The investigation showed that following the introduction of emission-reducing measures there was, at all locations, a highly significant reduction not only of all culturable indicator organisms (thermophilic actinomycetes and Aspergillus fumigatus) but also of total microorganism concentrations (p < 0,001). The introduction of the simple emission-reducing measures mentioned above, however, reduced the immission in the vicinity of the plant to such a degree that the natural background levels were reached at a distance of 150 m.     

Sequential coagulation factor VIIa domain binding to tissue factor

Vessel wall tissue factor (TF) is exposed to blood upon vascular damage which enables association with factor VIIa (FVIIa). This leads to initiation of the blood coagulation cascade through localization and allosteric induction of FVIIa procoagulant activity. To examine the docking pathway of the FVIIa-TF complex, various residues in the extracellular part of TF (sTF) that are known to interact with FVIIa were replaced with cysteines labelled with a fluorescent probe. By using stopped-flow fluorescence kinetic measurements in combination with surface plasmon resonance analysis, we studied the association of the resulting sTF variants with FVIIa. We found the docking trajectory to be a sequence of events in which the protease domain of FVIIa initiates contact with sTF. Thereafter, the two proteins are tethered via the first epidermal growth factor-like and finally the γ-carboxyglutamic acid (Gla) domain. The two labelled sTF residues interacting with the protease domain of FVIIa bind or become eventually ordered at different rates, revealing kinetic details pertinent to the allosteric activation of FVIIa by sTF. Moreover, when the Gla domain of FVIIa is removed the difference in the rate of association for the remaining domains is much more pronounced.     

Effects on the conformation of FVIIa by sTF and Ca binding: Studies of fluorescence resonance energy transfer and quenching

The apparent length of FVIIa in solution was estimated by a FRET analysis. Two fluorescent probes, fluorescein (Fl-FPR) and a rhodamine derivative (TMR), were covalently attached to FVIIa. The binding site of Fl-FPR was in the protease domain whereas TMR was positioned in the Gla domain, thus allowing a length measure over virtually the whole extension of the protein. From the FRET measurements, the distances between the two probes were determined to be 61.4 for free FVIIa and 65.5 Å for FVIIa bound to soluble tissue factor (sTF). These seemingly short distances, compared to those anticipated based on the complex crystal structure, require that the probes stretch towards each other. Thus, the apparent distance from the FRET analysis was shown to increase with 4 Å upon formation of a complex with sTF in solution. However, considering how protein dynamics, based on recent molecular dynamics simulations of FVIIa and sTF:FVIIa (Y.Z. Ohkubo, J.H. Morrissey, E. Tajkhorshid, J. Thromb. Haemost. 8 (2010) 1044–1053), can influence the apparent fluorescence signal our calculations indicated that the global average conformation of active-site inhibited FVIIa is nearly unaltered upon ligation to sTF.It is known from amidolytic activity measurements that Ca²⁺ binding leads to activation of FVIIa, but we have for the first time directly demonstrated conformational changes in the environment of the active site upon Ca²⁺ binding. Interestingly, this Ca²⁺-induced conformational change can be noted even in the presence of an inhibitor. Forming a complex with sTF further stabilized this conformational change, leading to a more inaccessible active-site located probe.     

Budding Yeast Greatwall and Endosulfines Control Activity and Spatial Regulation of PP2ACdc55 for Timely Mitotic Progression

Entry into mitosis is triggered by cyclinB/Cdk1, whose activity is abruptly raised by a positive feedback loop. The Greatwall kinase phosphorylates proteins of the endosulfine family and allows them to bind and inhibit the main Cdk1-counteracting PP2A-B55 phosphatase, thereby promoting mitotic entry. In contrast to most eukaryotic systems, Cdc14 is the main Cdk1-antagonizing phosphatase in budding yeast, while the PP2A^Cdc55 phosphatase promotes, instead of preventing, mitotic entry by participating to the positive feedback loop of Cdk1 activation. Here we show that budding yeast endosulfines (Igo1 and Igo2) bind to PP2A^Cdc55 in a cell cycle-regulated manner upon Greatwall (Rim15)-dependent phosphorylation. Phosphorylated Igo1 inhibits PP2A^Cdc55 activity in vitro and induces mitotic entry in Xenopus egg extracts, indicating that it bears a conserved PP2A-binding and -inhibitory activity. Surprisingly, deletion of IGO1 and IGO2 in yeast cells leads to a decrease in PP2A phosphatase activity, suggesting that endosulfines act also as positive regulators of PP2A in yeast. Consistently, RIM15 and IGO1/2 promote, like PP2A^Cdc55, timely entry into mitosis under temperature-stress, owing to the accumulation of Tyr-phosphorylated Cdk1. In addition, they contribute to the nuclear export of PP2A^Cdc55, which has recently been proposed to promote mitotic entry. Altogether, our data indicate that Igo proteins participate in the positive feedback loop for Cdk1 activation. We conclude that Greatwall, endosulfines, and PP2A are part of a regulatory module that has been conserved during evolution irrespective of PP2A function in the control of mitosis. However, this conserved module is adapted to account for differences in the regulation of mitotic entry in different organisms.