Control of yeast neutral trehalase by distinct polyphosphates and ribonucleic acid

Summary The activity of yeast trehalase when assayed at pH 7 in a crude extract was found to increase 2- to 3-fold upon incubation with 0.1 % (v/v) polyethyleneimine or other polycations such as polylysine (0.075-mMol) and calf thymus histories (0.08 mMol). Incubation with 3 mM-Mn²⁺ and 5 mM-Ca²⁺ also led to 3- and 1.6-fold increases in trehalase activity, respectively. The activities of 11 other enzymes assayed in the crude yeast extract did not increase after addition of polyethylene imine. At concentrations of polyethylenemine that maximally stimulated trehalase activity, 97% of the total RNA present in the crude extract, 40% of total protein, and 60% of the polyphosphate (assayed as inorganic phosphate liberated during 7 min incubation at 95 °C and pH O) were found to be precipitated. A similar finding was made with trehalase-stimulating concentrations of Mn²⁺. Activation of trehalase by polyethylene imine rendered this enzyme susceptible to inhibition by a preparation of total yeast RNA, inorganic polyphosphates, and related polyanions. We present further evidence that the removal of a distinct RNA and/or polyphosphate is the basic principle of polyetyleneimine-induced activation of trehalase.A more pronounced stimulation of trehalase activity (4-fold) could be obtained by enzymatic phosphorylation with ATP in the presence of cyclic AMP and Mg²⁺ as described by van Solingen and van der Plaat (1975) [9]. Thus, trehalase 2-fold activated by polyethylene imine was further activated by another 2-fold increase by enzymatic phosphorylation. Conversely, no additional stimulation by polyethylene imine was obtained for the maximally active, phosphorylated enzyme. We conclude that phosphorylation-mediated activation of trehalase is a two-step event, one being the removal of an inhibitor, which can be achieved by polyethylene imine or related cations independent on phosphorylation. The most likely candidate for the inhibitor appears to be a distinct RNA.

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Kontrolle der neutralen Hefen-Trehalase durch bestimmte Polyphosphate und RNA

Zusammenfassung Die in einem rohen Extrakt aus Hefe bei pH 7 gemessene Aktivität von Trehalase nimmt bei der Inkubation mit 0,1% Polyethylenimin oder mit anderen Polykationen, wie Polylysin (0,075 mmol) oder Histon aus Kalbsthymus (0,08 mmol) auf das zwei- bis dreifache zu. Auch die Inkubation mit 3 mmol Mn²⁺ oder 5 mmol Ca²⁺ führt zu einer 3-, bzw. 1,6fachen Zunahme der Trehalase-Aktivität. Die Aktivität von 11 anderen Enzymen, die im rohen Extrakt aus Hefe bestimmt wurden, nimmt nach Zusatz von Polyethylenimin nicht zu. Bei der Inkubation mit Polyethylenimin, das eine maximale Stimulierung der Trehalase-Aktivität bewirkt, werden 97% der gesamten Ribonucleinsäure, 40% des gesamten Proteins und 60% des Polyphosphats (bestimmt als anorganisches Phosphat, das in 7 min bei 95 °C und pH O freigesetzt wird) präcipitiert. Eine gleichartige Präcipitation wurde bei der Inkubation mit Trehalase-stimulierenden Konzentrationen von Mn²⁺ beobachtet. Mit Polyethylenimin aktivierte Trehalase wird durch Ribonucleinsäure aus Hefe, anorganisches Polyphosphat und verwandte Polyanionen gehemmt. Vermutlich führt die Entfernung einer bestimmten Ribonucleinsäure-Fraktion und/oder von Polyphosphat zu der Polyethylenimin-induzierten Aktivierung der Trehalase.Wie von van Solingen und van der Plaat 1975 beschrieben [9], wird eine 4fache Stimulierung von Trehalase durch enzymatische Phosphorylierung mit ATP in der Gegenwart von cyclischem AMP und Mg²⁺ erreicht. Das so durch Phosphorylierung maximal aktivierte Enzym kann durch Zusatz von Polyethylenimin nicht weiter aktiviert werden. Das mit Polyethylenimin 2fach aktivierte Enzym kann jedoch noch einmal 2fach weiter aktiviert werden durch enzymatische Phosphorylierung. Aus diesen Beobachtungen wird geschlossen, daß die durch enzymatische Phosphorylierung bewirkte Aktivierung der Trehalase ein zweistufiger Vorgang ist: Zuerst findet die Entfernung eines Inhibitors statt (dies kann auch mit Polyethylenimin oder mit verwandten Kationen unabhängig von der enzymatischen Phosphorylierung erreicht werden), hierauf folgt weitere Aktivierung durch Konformationsänderung des Enzyms. Wahrscheinlich handelt es sich bei dem durch Phosphorylierung bzw. Behandlung mit Polyethylenimin abgetrennten Inhibitor um eine gewisse Ribonucleinsäure-Fraktion.

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Abbreviation PEI polyethylene imine The followingenzymes are mentioned in the text: Alcohol dehydrogenase Glyceraldehyde-3-phosphate dehydrogenase (1.1.1.12), Malate dehydrogenase (1.1.1.37), Glucose-6-phosphate dehydrogenase (1.1.1.49), Glutamate dehydrogenase (NAD) (1.4.1.2), Glutamate dehydrogenase (NADP) (1.4.1.4), Aspartate -ketoglutarate aminotransferase (2.6.1.1), 6-Phosphofructokinase (2.7.1.11), Neutral trehalase (3.2.1.28), Pyruvate decarboxylase (4.1.1.1), Phosphoenolpyruvate carboxykinase (4.1.1.49).This paper is dedicated to Prof. Dr. Karl Decker on the occasion of his 60th birthday     

Funktionspolymere-ein interdisziplinäres forschungsgebiet-

Functional polymers differ from structural polymers in various aspects. The most evident difference is their lack of visibility as they are mostly empolyed in surface applications, e.g. as crystal growth modifiers to inhibit encrustation in detergents, as hair setting agents, or as electro-conductive films. Additional important differences concerning the motivation for new developments, market aspects, and customer relationships are the subject of this paper. The development of technology and market competence in the field of functional polymers and the special problems of low-volume, high-performance chemicals are discussed in detail. The paper deals with examples of recent developments: Polyaramides for membranes, 3,4-polyethylenedioxythiophene as an electroconductive polymer, polyaspartic acid as a biodegradable polycarboxylate in detergents, highly crosslinked, insoluble vinylpyrrolidone/vinylimidazole copolymers for the removal of heavy metals from wine, and finally an insoluble, crosslinked polyvinylpyrrolidone-iodine complex for the sterile filtration of liquids. The outlook discusses the research challenges which remain before a full understanding of the structure/property relationships can be attained.     

Reactive path integral quantum simulations of molecules solvated in superfluid helium

A hybrid ab initio   path integral molecular dynamics/bosonic path integral Monte Carlo simulation method has been developed, implemented and tested, which allows for the reactive simulations of molecules, clusters or complexes solvated by superfluid ⁴${}^4$He. The simulation takes into account “on-the-fly” the electronic structure and thus the chemical reactivity of the solutes, in conjunction with the Bose–Einstein statistics, and thus the superfluid character of this peculiar solvent. This enables investigations into cryochemical reactions taking place in helium nanodroplets, such as those used in helium nanodroplet isolation (HENDI) spectroscopy.     

Entrepreneurship and the carbon market: opportunities and challenges for South African entrepreneurs

The Kyoto Protocol and its implementation brought forward issues of climate change and its mitigation strategy by national measures through the creation of market mechanisms in carbon trading. The trading of emission certificates has become an important trade commodity worldwide, and its markets have diversified. While this opportunity has created new markets for entrepreneurs and actors that range from farmers to brokers, unequal involvement in most developing countries is noted. This has been mostly observed in those countries where entrepreneurship is often regarded as the cornerstone of economic growth and social improvement. South Africa has spearheaded other African countries in its implementation of Clean Development Mechanism (CDM) projects leading to carbon trading. Based on our research on South African entrepreneurship and its involvement in the carbon market, we conclude that albeit a number of opportunities, the biggest challenge for entrepreneurial participation in the carbon market remains in the nature and processes of CDM project implementation, the lack of a clear supportive system, limited access to financing and—more importantly—general ignorance of the trading opportunities by entrepreneurs. The complex nature of CDM projects themselves limits participation due to lack of the necessary skills on the national level leading to uneven distribution of CDM projects on provincial levels in South Africa. Recommendations are provided to overcome the obstacles and to promote entrepreneurial activity in the carbon market.     

Selective sterol transfer in the honey bee: Its significance and relationship to other hymenoptera

The honey bee,Apis mellifera, is one of only a few species of phytophagous insects known to be unable to convert C-24 alkyl phytosterols to cholesterol. Regardless of the dietary sterols available to worker bees, the major tissue sterol of brood reared by the workers is always 24-methylenecholesterol, followed by sitosterol and isofucosterol. Normally, little or no cholesterol is present in honey bee sterols. The maintenance of high levels of certain sterols is accomplished through a selective transfer of sterols from the endogenous sterol pools of the workers to the developing larvae through the brood food material secreted from the hypopharyngeal and mandibular glands and/or the honey stomach of the workers. The selective uptake and transfer of radiolabeled C₂₇, C₂₈ and C₂₉ sterols have been studied to correlate these aspects of sterol utilization with the discovery of an unusual molting hormone (ecdysteroid) in honey bee pupae as the major ecdysteroid of this stage of development. The phylogenetic implications of this selective transfer phenomenon in the honey bee and comparison with sterol metabolism in certain other hymenopteran species emphasize the diversity of steroid biochemistry in insects.     

A rapid method for the preparation of ganglioside Glac2 (GD3)

A method is described for the preparation of ganglioside G_lac2 [(II³(NeuAc)₂-LacCer, GD3] from cream of bovine milk using liquid-phase extraction with methanol or ethanol followed by anion exchange chromatography. The method is rapid and inexpensive; 1 kg cream, centrifuged from 14–15 L of bovine milk, yields approximately 70 mg of pure ganglioside G_lac2. The sialic acid constituent of ganglioside G_lac2 isolated from bovine milk cream consists solely of theN-acetylneuraminic acid derivative. The major components of its ceramide consist of octadecasphing-4-enine and the 22∶0 (behenic acid) and 23∶0 fatty acids. Short hand notations for gangliosides are according to Wiegandt (Ref. 1). G_lac2, (II³(NeuAc)₂-LacCer) is NeuAcα,8NeuAcα,-3Galβ,4Glcβ,1Cer; G_tet1, (II³NeuAc-Gg₄Cer) is Gal\,3GalNAcβ,-4(NeuAcα,3)Galβ,4Glcβ,1Cer.     

Improvement of Crystallinity of KZr2P3O12 by Sol-Gel Synthesis

The X-ray diffraction pattern for potassium-zirconium phosphate is significantly improved when the material is prepared by the sol-gel route rather than the powder method. Peaks at high, low, and intermediate angles are presented and compared for the two methods. It is assumed that the more nearly homogeneous mixing of the elements in the sol-gel process is responsible for the significant improvement in crystallinity.     

Open annotations on multimedia Web resources

Many Web portals allow users to associate additional information with existing multimedia resources such as images, audio, and video. However, these portals are usually closed systems and user-generated annotations are almost always kept locked up and remain inaccessible to the Web of Data. We believe that an important step to take is the integration of multimedia annotations and the Linked Data principles. We present the current state of the Open Annotation Model, explain our design rationale, and describe how the model can represent user annotations on multimedia Web resources. Applying this model in Web portals and devices, which support user annotations, should allow clients to easily publish and consume, thus exchange annotations on multimedia Web resources via common Web standards.     

A framework for improved video text detection and recognition

Text displayed in a video is an essential part for the high-level semantic information of the video content. Therefore, video text can be used as a valuable source for automated video indexing in digital video libraries. In this paper, we propose a workflow for video text detection and recognition. In the text detection stage, we have developed a fast localization-verification scheme, in which an edge-based multi-scale text detector first identifies potential text candidates with high recall rate. Then, detected candidate text lines are refined by using an image entropy-based filter. Finally, Stroke Width Transform (SWT)- and Support Vector Machine (SVM)-based verification procedures are applied to eliminate the false alarms. For text recognition, we have developed a novel skeleton-based binarization method in order to separate text from complex backgrounds to make it processible for standard OCR (Optical Character Recognition) software. Operability and accuracy of proposed text detection and binarization methods have been evaluated by using publicly available test data sets.