Accumulation of nitrite and sulfite in yeast cells and synergistic depletion of the intracellular ATP content

Summary When nitrite or sulfite are applied to yeast cells below pH 5.0, an enormous intracellular accumulation occurs. It is assumed that nitrite and sulfite penetrate the cell membrane in their undissociated forms as nitrous acid (pK = 3.3) or sulfurous acid (pK =1.8), respectively. Due to the neutral intracellular pH they are trapped inside the cell in their anionic forms, which are impermeable to the cell membrane. It has previously been shown that sulfite causes a rapid depletion of the ATP content of yeast cells [Schimz, K. L. and Holzer, H. (1979) resp. Hinze et al. as above]. Similarly, millimolar concentrations of nitrite decrease the ATP level to less than 10% of the initial value. Nitrite and sulfite in combination deplete the ATP content of yeast cells much stronger than expected for the sum of the separate effects of these compounds (synergistic effect).

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Akkumulation von Nitrit und Sulfit in Hefezellen und synergistischer Abfall des intrazellulären ATP-Gehalts

Zusammenfassung Wenn Hefezellen mit Nitrit oder Sulft bei pH-Werten unter 5,0 inkubiert werden, beobachtet man eine starke intracelluläre Akkumulation von Nitrit, bzw. Sulfit. Es ist anzunehmen, daß Nitrit und Sulfit in ihrer undissoziierten Form als salpetrige Säure (pK=3,3) bzw. schweflige Säure (pK=1,8) penetrieren und dann in den Zellen durch Neutralisation zu den anionischen Formen, die die Zellmembran nicht mehr permeieren können, abgefangen werden. Ähnlich dem früher beschriebenen raschen Abfall des ATP-Gehaltes nach Zusatz von Sulfit in Hefe [Schimz, KL und Holzer H (1979) Arch Microbiol 121:225–229] und in Bakterien [Hinze H, Maier K, Holzer H (1981) Z Lebensm Unters Forsch 172:389-392] verursacht auch Nitrit einen raschen Abfall des ATP-Gehaltes in Hefe auf weniger als 10 % des Anfangswertes. Werden Nitrit und Sulfit in Kombination verabreicht, so beobachtet man einen wesentlich stärkeren Abfall des ATP-Gehaltes als er aus der Summe der Einzeleffekte von Nitrit, bzw. Sulfit zu erwarten wäre (Synergistischer Effekt).

     

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     

Role of Macrophages and RhoA Pathway in Atherosclerosis

The development, progression, or stabilization of the atherosclerotic plaque depends on the pro-inflammatory and anti-inflammatory macrophages. The influx of the macrophages and the regulation of macrophage phenotype, inflammatory or anti-inflammatory, are controlled by the small GTPase RhoA and its downstream effectors. Therefore, macrophages and the components of the RhoA pathway are attractive targets for anti-atherosclerotic therapies, which would inhibit macrophage influx and inflammatory phenotype, maintain an anti-inflammatory environment, and promote tissue remodeling and repair. Here, we discuss the recent findings on the role of macrophages and RhoA pathway in the atherosclerotic plaque formation and resolution and the novel therapeutic approaches.     

Analytical studies on the temperature distribution in spiral plate heat exchangers: Straightforward design formulae for efficiency and mean temperature difference

The temperature distribution in spiral plate heat exchangers has been calculated numerically to obtain the efficiency and the logarithmic mean temperature difference (LMTD) correction factors F as a function of the number of transfer units N, the number of turns n, and the heat capacity rate ratio C. It has been found that the LMTD correction factors, when plotted against the number of transfer units per turn N/n, fall approximately on a single curve. That curve for balanced countercurrent operation (C = −1) can be very closely represented by the simple formula F = (n/N)tanh(N/n). From that simple analytic representation of our numerical results it was concluded that a simpler physical model might exist to represent the overall behaviour of a spiral plate heat exchanger equally well. In fact, a countercurrent cascade of n cocurrent heat exchangers does result exactly in the above-mentioned formula for the LMTD correction factor. From that model the F-factors for other heat capacity rate ratios C (−1 < C 0) can also be calculated and they are in sufficient agreement with the numerical results.     

The oxidation and self-heating of metal sulphides as an electrochemical corrosion phenomenon

Experiments with a representative number of technically interesting metal sulphides trace out hydrogen peroxide as the reactive intermediate of sulphide oxidation. It is formed through an electrochemical reduction of molecular oxygen. The formation of sulphur or sulphate as the final state of oxidation is determined by consecutive reactions which are critically dependent on the catalytical properties of the surface and the solution for hydrogen peroxide decomposition. The individual steps of the reaction are discussed in some detail. Conclusions with respect to a possible control of the autogeneous heating are drawn.     

Influence of different process and material parameters on chemical foaming of fluorinated ethylene propylene copolymers

In this article, the chemical foaming of fluorinated ethylene propylene copolymers (FEP) is investigated. For the laboratory scale foam extrusion process, a chemical blowing agent adapted to the high melting temperature of FEP had to be found. Foaming experiments were carried out varying process and material parameters. Foam densities as well as cellular structures were analyzed to characterize how the parameters influence the foaming behavior of the FEP melt. An increasing rate of the foam extrusion caused an augmentation of cell numbers at a simultaneous decrease of their diameters. Moreover, a pronounced reduction of the foam density with increasing output rate could be observed. Changing the temperature of the foam extrusion, an optimum in density reduction and homogeneity of the cell size was found. Furthermore, the influence of an appropriate nucleation agent on the resulting geometry and amount of cells was investigated. Though the chemical blowing agent itself can act as an implement for nucleating cell growth, an addition of 10 wt% of calcium fluoride particles showed a significant augmentation in cell quantity. For the chemical foam extrusion process, three FEP with different viscosities were characterized. With decreasing viscosity, a reduction in foam density could be observed for all temperatures. POLYM. ENG. SCI., 47:1740–1749, 2007. © 2007 Society of Plastics Engineers     

Scandinavian Thins on Top of Cake: New and Improved Algorithms for Stacking and Packing

We show how to compute the smallest rectangle that can enclose any polygon, from a given set of polygons, in nearly linear time; we also present a PTAS for the problem, as well as a linear-time algorithm for the case when the polygons are rectangles themselves. We prove that finding a smallest convex polygon that encloses any of the given polygons is NP-hard, and give a PTAS for minimizing the perimeter of the convex enclosure. We also give efficient algorithms to find the smallest rectangle simultaneously enclosing a given pair of convex polygons.