The occurrence of peptaibols and structurally related peptaibiotics in fungi and their mass spectrometric identification via diagnostic fragment ions.

Peptaibols and related peptide antibiotics (peptaibiotics) display diagnostically useful fragmentation patterns during mass spectrometry (FAB-MS, ESI-CID-MS/MS and CID-MSn]. The paper compiles fragmentation data of pseudo-molecular ions reported in the literature as a guide to the rational identification of recurrently isolated and new peptaibols and peptaibiotics. Taxonomic and ecological aspects of microorganisms producing peptaibols and peptaibiotics are discussed.     

EFFECTS OF SINGLE AND MULTIPLE EXPOSURES OF FERULIC ACID ON THE VEGETATIVE AND REPRODUCTIVE GROWTH OF PHASEOLUS VULGARIS BBL-290

Phaseolus vulgaris BBL-290 plants were grown in growth chambers in the Southeastern Plant Environment Laboratory and exposed to either single (at seedling, flower, or podfill) or multiple (biweekly or weekly) treatments of ferulic acid (FA). In the first experiment, plants were harvested one week after FA treatment (0, 1.0, 2.0 mM) and at final harvest (56 days old). FA delayed leaf expansion during the seedling and flowering stages. The total plant leaf area and the plant dry weight of plants treated with 1.0 and 2.0 mM FA as seedlings were reduced one week after treatment by 38–48%. The total plant leaf area and the plant dry weight of plants treated at flowering with 2.0 mM FA were reduced by 25% one week after treatment. Treatment with 2.0 mM FA at podfill caused the senescence and abscission of older leaves and reduced total plant leaf area, plant dry weight and mean pod dry weight by 54, 40, and 48%, respectively, one week after treatment. The plants treated at the seedling and flowering stages recovered by final harvest. In a subsequent experiment, FA (0, 0.50, 1.0, 1.5 mM) reduced total plant leaf area at the seedling and flowering stages but not at podfill. The youngest expanding leaves were most sensitive to FA at flowering. The leaf area of these leaves was reduced by 35 and 25%, one and two weeks after treatment, respectively. Their absolute growth rates were reduced from 31 to 56% one week after treatment at flowering. Their relative growth rates were reduced by 50% one week after treatment. Growth rates then recovered within two weeks after treatment. In the final experiment, biweekly exposures of FA (0.25, 0.50, 0.75, 1.0) reduced total plant leaf area but did not affect any other growth parameters. Weekly exposures of FA (0.25, 0.50, 0.75, 1.0) reduced total plant leaf area up to 34%, absolute growth rate up to 58%, leaf number up to 31% and pod number up to 58%. As the frequency of exposure to FA increased, the concentration necessary to affect bean plant growth and development decreased.     

Stofftransport im Ovar von Pyrrhocoris apterus L.

Zusammenfassung Synthese und Transport von DNS, RNS und Proteinen in einem telotrophen Ovar wurden autoradiographisch untersucht. Das Vorkommen von Endomitosen in Kernen sich differenzierender Nährzellen und des Follikelepithels können für Pyrrhocoris bestätigt werden, desgleichen der Transport von hochmolekularer RNS von den Nährzellkernen zu den Oocyten in der Phase euplasmatischen Wachstums. Ein Transport in jüngere Oocyten erfolgt nicht. Ein Beitrag des Follikelepithels zur RNS-Versorgung der Oocyten wie er für eine andere Wanze beschrieben wurde, ist bei Pyrrhocoris nicht feststellbar; gleichfalls im Gegensatz zu den Befunden an anderen Wanzen findet eine relativ schwache RNS-Synthese in allen Oocytenkernen statt. Protein wird mit unterschiedlicher Intensität in der gesamten Ovariole synthetisiert und von den Nährzellen in die Oocyten transportiert.Wie im polytrophen Ovar sind zwei RNS-Transporte unterscheidbar: Die langsamere Hauptfraktion hat anscheinend die gleiche, geringe Geschwindigkeit wie der Proteintransport; es wird daher angenommen, daß es sich um die passive Verfrachtung von Ribosomen im Strom des Euplasma handelt. Eine geringere Menge markierter RNS wird schneller durch das Cytoplasma transportiert und als messenger-RNS diskutiert.

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Summary Synthesis and transport of DNA, RNA, and proteins were studied in a telotrophic ovary using autoradiography. The occurrence of endomitosis in nuclei of differentiating nurse cells and follicular epithelium can be confirmed for Pyrrhocoris and also the transport of high molecular RNA from the nurse cell nuclei into the oocytes during their phase of euplasmatic growth. There is no transport into younger oocytes. In Pyrrhocoris no contribution of the follicular epithelium to the RNA supply of the oocytes is found as described for a different Heteroptere; contrary to the findings reported for other Heteroptera, a relatively low rate of RNA synthesis is detected in all oocyte nuclei. Protein is synthesized with different intensity in all parts of the ovariole and transported from the nurse cells into the oocytes.It is possible, like in the polytrophic ovary, to distinguish between two modes of RNA transport. The bulk of the RNA seems to moove at the same low speed as the proteins, presumably representing a passive movement of ribosomes in the flow of euplasma. A smaller fraction of labelled RNA is transported faster through the cytoplasm; it may consist of messenger RNA.

Die Untersuchungen wurden z.T. mit Mitteln durchgeführt, die die Deutsche Forschungsgemeinschaft Herrn Prof. Bier zur Verfügung stellte.     

Ultrastructural studies of oogenesis in some european amphibians

Summary Oogenesis was studied in adult Triturus vulgaris (Urodela) with the electron microscope. The oocytes investigated ranged between 50 m and 1600 m in diameter.Two types of yolk platelet formation were found. Since both types involve the incorporation of high numbers of pinocytotic vesicles they are believed to be of an extraoocytic origin. On the basis of the order of their appearance they were named primary and secondary yolk.Five different types of vesicles were found, which participate in a variety of activities, such as yolk formation and the formation of the Golgi apparatus. They originate from four different sources, namely the nuclear membrane, the cytoplasm in connection with ribosome-like particles, the Golgi apparatus and the plasma membrane through pinocytosis. The results obtained were discussed especially with respect to differences found between the anura and the urodela, such as the presence or absence of cortical granules or equivalent structures.The authors wish to thank Prof. Dr. K.S. Ludwig for his valuable criticism and encouragement during the course of this study, Messrs. H. Boffin, C. Evers and Miss D. Lovri for their capable technical assistance.     

One pot synthesis of GDP‐mannose by a multi‐enzyme cascade for enzymatic assembly of lipid‐linked oligosaccharides

Glycosylation of proteins is a key function of the biosynthetic‐secretory pathway in the endoplasmic reticulum (ER) and Golgi apparatus. Glycosylated proteins play a crucial role in cell trafficking and signaling, cell‐cell adhesion, blood‐group antigenicity, and immune response. In addition, the glycosylation of proteins is an important parameter in the optimization of many glycoprotein‐based drugs such as monoclonal antibodies. In vitro glycoengineering of proteins requires glycosyltransferases as well as expensive nucleotide sugars. Here, we present a designed pathway consisting of five enzymes, glucokinase (Glk), phosphomannomutase (ManB), mannose‐1‐phosphate‐guanyltransferase (ManC), inorganic pyrophosphatase (PmPpA), and 1‐domain polyphosphate kinase 2 (1D‐Ppk2) expressed in E. coli for the cell‐free production and regeneration of GDP‐mannose from mannose and polyphosphate with catalytic amounts of GDP and ADP. It was shown that GDP‐mannose is produced at various conditions, that is pH 7–8, temperature 25–35°C and co‐factor concentrations of 5–20 mM MgCl₂. The maximum reaction rate of GDP‐mannose achieved was 2.7 μM/min at 30°C and 10 mM MgCl₂ producing 566 nmol GDP‐mannose after a reaction time of 240 min. With respect to the initial GDP concentration (0.8 mM) this is equivalent to a yield of 71%. Additionally, the cascade was coupled to purified, transmembrane‐deleted Alg1 (ALG1ΔTM), the first mannosyltransferase in the ER‐associated lipid‐linked oligosaccharide (LLO) assembly. Thereby, in a one‐pot reaction, phytanyl‐PP‐(GlcNAc)₂‐Man₁ was produced with efficient nucleotide sugar regeneration for the first time. Phytanyl‐PP‐(GlcNAc)₂‐Man₁ can serve as a substrate for the synthesis of LLO for the cell‐free in vitro glycosylation of proteins. A high‐performance anion exchange chromatography method with UV and conductivity detection (HPAEC‐UV/CD) assay was optimized and validated to determine the enzyme kinetics. The established kinetic model enabled the optimization of the GDP‐mannose regenerating cascade and can further be used to study coupling of the GDP‐mannose cascade with glycosyltransferases. Overall, the study envisages a first step towards the development of a platform for the cell‐free production of LLOs as precursors for in vitro glycoengineering of proteins.     

Research Design and the Politics of Abstraction: Unpacking the Environmentality of Scientific Practice in Socioecological Assessments

Scientific assessments of socio-ecological systems are becoming mainstays in guiding policymaking and other interventions in response to global environmental change. The environmentality literature emphasizes the institutional architecture of emergent science-policy regimes and how scientific research is used in political settings, creating new modes of governance and subjectivities. However, there has been relatively little attention to domain-level socio-ecological assessments as socially produced technologies where specific scientific choices are mechanisms connecting governance architecture and popular subjectivities.Combining empirical case study and literature review, assessment technologies are analyzed in three domains: vulnerability assessment, ecosystem services assessment, life-cycle assessment. Using conceptualization, operationalization, and institutionalization as analytical lenses, the cases illustrate ways that scientific choices simplify complex socio-ecological relationships with implications for both governance practices and subjectivities. Furthermore, findings explore the possibility for assessments to be more inclusive of diverse social values and practices, enabling more empowering subjectivities.     

Galectin-1 Overexpression in Endometriosis and Its Regulation by Neuropeptides (CRH,UCN) Indicating Its Important Role in Reproduction and Inflammation

Endometriosis is an inflammatory disease of women of reproductive age featured by the presence of ectopic endometrium and is strongly related to infertility. Galectins, carbonhydrate-binding proteins, have been found to have pro- or anti-inflammatory roles in the reproductive tract and in pathological conditions concerning infertility. Galectin-1, which is expressed at endometrium and decidua, plays a major role in implantation and trophoblast invasion. Also, the neuropeptides, corticotropin releasing hormone (CRH) and urocortin (UCN) and their receptors are expressed in eutopic and ectopic endometrium showing a differential expression pattern in endometriotic women compared to healthy ones. The aim of this study was to examine the galectin-1 expression in endometriotic lesions and compare its expression in eutopic endometrium of endometriotic and healthy women. Furthermore, we examined the effect of CRH and UCN in galectin-1 expression in Ishikawa cell line and macrophages and investigated the implication of CRHR1 in these responses. Eutopic and ectopic endometrium specimens, Ishikawa cell line and mice macrophages were used. Immunohistochemistry and western blot analysis were performed in order to identify galectin-1 expression in ectopic and eutopic endometrium of women with and without endometriosis and the regulatory effect of CRH and UCN on galectin-1 expression. This study presents for the first time that galectin-1 is overexpressed in endometriotic lesions compared to eutopic endometrium of endometriotic women and is more abundantly expressed in eutopic endometrium of disease women compared to healthy ones. Furthermore, it is shown that CRH and UCN upregulate galectin-1 expression in Ishikawa cell line and macrophages and this effect is mediated through CRHR1. These results suggest that galectin-1 might play an important role in endometriosis pathology and infertility profile of women suffering from endometriosis by being at the same time regulated by CRH and UCN interfering in the immune disequilibrium which characterizes this pathological condition.