6-Phosphofructo-2-kinase and D-fructose-2,6-bisphosphatase activities in mung bean seedlings

6-Phosphofructo-2-kinase (ATP: D-fructose-6-phosphate-2-phosphotransferase) and D-fructose-2,6-bisphosphatase activities have been found in extracts prepared from etiolated mung bean seedlings. The activity of 6-phosphofructo-2-kinase exhibits a sigmoidal shape in response to changes in concentrations of both substrates, D-fructose 6-phosphate and ATP (S_0.5 values of 1.8 and 1.2 mM, respectively). Inorganic orthophosphate (P_i) has a strong stimulating effect on the 2-kinase activity (A_0.5 at about 2 mM), moderately increasing the V_max and modifying the response into hyperbolic curves with K_m values of 0.4 and 0.2 mM for fructose 6-phosphate and ATP, respectively. 3-Phosphoglycerate (I_0.5 about 0.15 mM) partially inhibited the kinase activity by counteracting the P_i activation. In contrast, the activity of D-fructose-2,6-bisphosphatase (K_m 0.38 mM) is strongly inhibited by P_i (I_0.5 0.8 mM) lowering its affinity to fructose-2,6-P₂ (K_m 1.4 mM). 3-Phosphoglycerate activites the enzyme (A_0.5 at about 0.3 mM) without causing a significant change in its K_m for fructose-2,6-P₂. The activities of both of these enzymes in relationship to the metabolic role of D-fructose 2,6-bisphosphate in the germinating seed is discussed.     

Total Internal Reflection Accounts for the Bright Color of the Saharan Silver Ant

The Saharan silver ant Cataglyphis bombycina is one of the terrestrial living organisms best adapted to tolerate high temperatures. It has recently been shown that the hairs covering the ant’s dorsal body part are responsible for its silvery appearance. The hairs have a triangular cross-section with two corrugated surfaces allowing a high optical reflection in the visible and near-infrared (NIR) range of the spectrum while maximizing heat emissivity in the mid-infrared (MIR). Those two effects account for remarkable thermoregulatory properties, enabling the ant to maintain a lower thermal steady state and to cope with the high temperature of its natural habitat. In this paper, we further investigate how geometrical optical and high reflection properties account for the bright silver color of C. bombycina. Using optical ray-tracing models and attenuated total reflection (ATR) experiments, we show that, for a large range of incidence angles, total internal reflection (TIR) conditions are satisfied on the basal face of each hair for light entering and exiting through its upper faces. The reflection properties of the hairs are further enhanced by the presence of the corrugated surface, giving them an almost total specular reflectance for most incidence angles. We also show that hairs provide an almost 10-fold increase in light reflection, and we confirm experimentally that they are responsible for a lower internal body temperature under incident sunlight. Overall, this study improves our understanding of the optical mechanisms responsible for the silver color of C. bombycina and the remarkable thermoregulatory properties of the hair coat covering the ant’s body.     

Adipose Tissue-Derived Stem Cells Reduce Acute and Chronic Kidney Damage in Mice

Acute and chronic kidney injuries (AKI and CKI) constitute syndromes responsible for a large part of renal failures, and are today still associated with high mortality rates. Given the lack of more effective therapies, there has been intense focus on the use stem cells for organ protective and regenerative effects. Mesenchymal stem cells (MSCs) have shown great potential in the treatment of various diseases of immune character, although there is still debate on its mechanism of action. Thus, for a greater understanding of the role of MSCs, we evaluated the effect of adipose tissue-derived stem cells (AdSCs) in an experimental model of nephrotoxicity induced by folic acid (FA) in FVB mice. AdSC-treated animals displayed kidney functional improvement 24h after therapy, represented by reduced serum urea after FA. These data correlated with cell cycle regulation and immune response modulation via reduced chemokine expression and reduced neutrophil infiltrate. Long-term analyses, 4 weeks after FA, indicated that AdSC treatment reduced kidney fibrosis and chronic inflammation. These were demonstrated by reduced interstitial collagen deposition and tissue chemokine and cytokine expression. Thus, we concluded that AdSC treatment played a protective role in the framework of nephrotoxic injury via modulation of inflammation and cell cycle regulation, resulting in reduced kidney damage and functional improvement, inhibiting organ fibrosis and providing long-term immune regulation.     

Kofyar cash-cropping: Choice and change in indigenous agricultural development

Amid discussions of an agricultural crisis and the failure of largescale, mechanized, capitalintensive development schemes in Nigeria, the Kofyar of Plateau State provide a case study of farmers spontaneously expanding food crop production for the market, using indigenous lowenergy technology. Temporary, followed by permanent, migration from the Jos Plateau homeland to frontier settlements on the fertile Benue plains has been accompanied by a change from initial shifting cultivation in forest clearings to permanent, intensively tilled and fertilized homestead fields. Labor is organized primarily in households that have grown in size and complexity. Cooperative and exchange work groups are also important for meeting seasonal bottlenecks and providing the careful, disciplined cultivation that intensive agriculture requires. Kofyar now devote up to 50% of their labor to cash crops, and they purchase considerable quantities of manufactured goods and medical services. Their uncoerced adaptation to an environment of new land resources and market incentives suggests both the advantages of indigenous development with a minimum of state control or interference and the limitations of a conventional dependency theory perspective.     

A sodium dodecyl sulfate-gradient gel electrophoresis system that separates polypeptides in the molecular weight range of 1500 to 100,000

A sodium dodecyl sulfate-polyacrylamide gradient gel system is described. It combines a linear gradient in polyacrylamide from 10.2 to 30.2% in the separating gel and the discontinuous ammediol/glycine buffer system suggested by Bury (A. F. Bury, 1981, J. Chromatogr. 213, 491-500). This urea-free electrophoretic system provides high resolution and clean separation patterns of proteins and polypeptides with molecular weights from 1500 to 100,000. It is especially suited for studying complex mixtures of proteins and proteolytic fragments, in particular with regard to immunoelectrotransfer blot techniques.     

Changes in membrane potential and resistance caused by transient increase of potassium conductance in the unicellular green alga Eremosphaera viridis

The electrophysiological membrane parameters of the unicellular green alga Eremosphaera viridis were determined using an improved computer-supported single-microelectrode technique. These cells developed an average membrane potential of-150 mV in the light and a specific resistance of 1 Ω m² with an external potassium concentration of 1.1 mM and pH 5.5. In the dark, many cells showed a less polarized potential of 30–40 mV and a smaller membrane resistance. At potassium concentrations in the external medium higher than 1 mM, the membrane potential strongly depends on the external potassium content apart from a small electrogenic component. At concentrations lower than 1 mM K⁺, a dependence of the membrane potential upon external potassium concentrations could not be verified. Inserting the internal ion activities in the Goldmann equation shows that, in this range, the proton conductance seems to be predominant over the potassium conductance. Transient changes in the membrane potential and in the membrane resistance were observed after switching off the light, after addition of 3-(3′,4′-dichlorophenyl)-1,1-dimethylurea or N,N′-dicyclohexylcarbodiimide, after a sudden decrease in temperature, and after current pulses. These changes resemble the action potentials (AP) found in other plant cells (Chara, Acetabularia). On average, the AP has a delay period of 5.1 s and a duration of 43.8 s showing a sudden decrease and a slower regeneration. The voltage peak during an AP followed exactly the Nernst potential of potassium over a range of external potassium concentrations from 5 μM to 0.2 M. This is true for depolarization or hyperpolarization, depending on the external K⁺-concentration. Tetraethylammonium-hydrogensulphate, a rather specific inhibitor of K⁺ channels in nervous cells, suppressed the AP. The correlation of the appearance of the AP with a short-term opening of potassium channels in the membrane of Eremosphaera is discussed.     

Über den Einfluß der Vorbelichtung auf die anschließende Phosphorylierung im Dunkeln bei einzelligen Grünalgen (Ankistrodesmus braunii)

Zusammenfassung Wird einzelligen Algen (Ankistrodesmus braunii) nach Vorbelichtung in anschließender Dunkelheit ³²P-markiertes Phosphat geboten, so tritt gegenüber Dauerdunkel eine erhebliche Förderung der ³²P-Einlagerung auf. Die nach Vorbelichtung bestimmte Markierung der aufgetrennten Phosphatfraktionen ähnelt sehr derjenigen im Dauerlicht. Die erhöhte Dunkelphosphorylierung nach Vorbelichtung hängt von der CO₂-Konzentration, von der Lichtintensität und der Zeit der Vorbelichtung ab. Unter den vorliegenden Bedingungen waren 7 min Vorbelichtung zur maximalen Förderung nötig. Die Halbwertzeit des Abklingens betrug etwa 4 min.Aus den Experimenten geht hervor, daß durch die Belichtung der Algen auch in vivo ein Zustand gebildet wird, der noch nach Belichtung eine Zeitlang im Dumkeln eine Erhöhung der ³²P-Einlagerung erlaubt.Es wird diskutiert, ob es sich einerseits um die Bildung einer im Licht reduzierten Substanz R handeln könnte, die für eine begrenzte Zeit in Dunkelheit noch einen cyclischen, mit Phosphorylierung gekoppelten Elektronentransport aufrechterhalten kann. Andererseits könnte durch die Vorbelichtung ein energiereiches Zwischenprodukt X _E — oder auch ein Protonenpool — gebildet werden, das bei dem Energietransfer vom Elektronentransportsystem zur ATP aufgebaut wird. Schließlich muß berücksichtigt werden, daß durch die Vorbelichtung an den Chloroplastenmembranen ein verstärkter ATP-Pi-Austausch zustande kommen könnte, der nach Belichtung nur langsam abklingt.

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Influence of preillumination on subsequent phosphorylation in the darkness of unicellular green algae (Ankistrodesmus braunii)

Summary Preilluminated unicellular green algae (Ankistrodesmus braunii) were treated in the subsequent darkness with ³²PO₄. The post-illumination dark incorporation was considerably increased compared with the control in continuous dark. The labeling of the separated phosphate-fractions was similar to that of continuous light. The light-induced dark incorporation depended from the light intensity as well as from the time of preillumination. A preillumination of 7 min was required for a maximal enhancement of this preillumination effect. On the other hand the effect diminished in darkness with a half life of approximately 4 min. Finally the enhancement was found to be greater in the absence of CO₂ than in the presence of CO₂.The experiments demonstrate the light-induced formation of a state in the algae, which permits the enhancement of ³²P-incorporation into several phosphate-fractions for a limited time during subsequent darkness.It is discussed, that this may be performed through the formation of a light-reduced substance R maintaining for a limited time a cyclic electron transport in darkness, coupled with phosphorylation. On the other hand it seems possible, that preillumination induces a high energy intermediate X _E—this could also be a pool of protons—formed in the course of energy-transfer from electron transport to ATP-formation. But we must consider also the possibility that light accellerates the ATP-Pi exchange on chloroplast-membranes for a time after preillumination.

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Abkürzungen ATP Adenosintriphosphat - ADP Adenosindiphosphat - Pi Orthophosphat - Poly-P anorganisches Polyphosphat - RNS Ribonucleinsäure - TCE Trichloressigsäure - 2,4-DNP 2,4-Dinitrophenol Stipendiat der Nishina-Gedächtnis-Stiftung (Japan) für 1963.