Free and polymerized tubulin in cultured bone cells and Chinese hamster ovary cells: the influence of cold and hormones

A low pH method of liposome-membrane fusion (Schneider et al., 1980, Proc. Natl. Acad. Sci. U. S. A. 77:442) was used to enrich the mitochondrial inner membrane lipid bilayer 30-700% with exogenous phospholipid and cholesterol. By varying the phospholipid-to- cholesterol ratio of the liposomes it was possible to incorporate specific amounts of cholesterol (up to 44 mol %) into the inner membrane bilayer in a controlled fashion. The membrane surface area increased proportionally to the increase in total membrane bilayer lipid. Inner membrane enriched with phospholipid only, or with phospholipid plus cholesterol up to 20 mol %, showed randomly distributed intramembrane particles (integral proteins) in the membrane plane, and the average distance between intramembrane particles increased proportionally to the amount of newly incorporated lipid. Membranes containing between 20 and 27 mol % cholesterol exhibited small clusters of intramembrane particles while cholesterol contents above 27 mol % resulted in larger aggregations of intramembrane particles. In phospholipid-enriched membranes with randomly dispersed intramembrane particles, electron transfer activities from NADH- and succinate-dehydrogenase to cytochrome c decreased proportionally to the increase in distance between the particles. In contrast, these electron- transfer activities increased with decreasing distances between intramembrane particles brought about by cholesterol incorporation. These results indicate that (a) catalytically interacting redox components in the mitochondrial inner membrane such as the dehydrogenase complexes, ubiquinone, and heme proteins are independent, laterally diffusible components; (b) the average distance between these redox components is effected by the available surface area of the membrane lipid bilayer; and (c) the distance over which redox components diffuse before collision and electron transfer mediates the rate of such transfer.     

Early conception factor lateral flow assays for pregnancy in the mare

The ECF™ lateral flow assay test is marketed to detect non-pregnancy in mares. The objectives of the present study were to determine the accuracy of the ECF test, the accuracy of the electronic reader accompanying the ECF test, and agreement between two human readers and the electronic reader. Serum samples were collected from anestrus, cycling but not inseminated, and inseminated mares, and were evaluated with the ECF™ test (EDP Biotech Company, Knoxville, TN, USA) at The Ohio State University and at the EDP Biotech Laboratory. Specificity ranged from 0.07 to 0.16, the negative predictive value ranged from 0.15 to 0.33, and accuracy ranged from 0.43 to 0.52. The electronic reader did not add improve the accuracy or predictive values of the test. Based on the electronic reader, 80.0% of the serum samples collected from the anestrus mares were false positives; Readers 1 and 2 had 60.0 and 33.3% false positives, respectively. For samples collected during the estrous cycle, 83.9% were false positives by the electronic reader, whereas Readers 1 and 2 had 43.7 and 26.4% false positives. We concluded that, regardless of whether the test strips were evaluated by a human or electronic reader, this assay was not accurate for determination of the non-pregnant mare.     

Fluorescent energy transfer measurements on fluorescein isothiocyanate modified cytochrome P-450 LM2

The distance between the heme iron and the N-terminus of cytochrome P-450 LM2 was determined by fluorescence energy transfer measurements. Fluorescein isothiocyanate which was covalently bound to the N-terminal methionine was used as donor chromophor. The Ro value between fluorescein isothiocyanate and the heme was calculated to be 3.98 nm. The distance between the nitrogen of the N-terminal methionine and the heme was estimated with 2.84 +/- 0.23 nm excluding most likely the N-terminal amino acid of cytochrome P-450 LM2 to participate in the electron transfer to the heme iron. A cytochrome P-450 LM2 membrane model is proposed.     

Über die gesteigerte Peroxydaseaktivität in gestörtenPhaseolus-Artbastarden

Ohne ZusammenfassungMit 3 Textabbildungen     

Untersuchungen über die Ursachen der Peroxydasevermehrung in gestörtenPhaseolus-Artbastarden

Zusammenfassung Um Aufschluß darüber zu erlangen, wie die Vermehrung der Peroxydase in gestörtenPhaseolus-Artbastarden (Phaseolus vulgaris x Phaseolus coccineus) zustande kommt, wurde die Fähigkeit der Bastarde zur Bildung von Peroxydaseapoenzym und Peroxydasecoenzym (=Hämatin) geprüft. Diese Prüfung wurde auf Grund des Befundes vonTheorell u. Mitarb., daß sich Peroxydaseapo- und Peroxydasecoenzym beiph-Werten um 7 spontan zur aktiven Peroxydase vereinigen, vorgenommen.Zusatz von Peroxydaseapoenzym zu Blatthomogenaten normaler und gestörter Pflanzen beeinflußt in keinem Fall die Peroxydasereaktion. Coenzymzusatz bleibt wirkungslos bei normalen, führt aber zu einem starken Anstieg der Peroxydasereaktion bei gestörten Bastarden. Wie der Chlorophyllgehalt ist auch der Gesamthämatingehalt bei den gestörten Bastarden stark vermindert.Aus den Befunden darf geschlossen werden, daß normale und gestörte Bastarde kein freies Hämatin enthalten und daß freies Apoenzym den normalen Bastarden fehlt, bei den gestörten Bastarden aber in größerer Menge vorhanden ist. Bei den normalen Bastarden sind Apound Coenzymproduktion aufeinander abgestimmt, während bei den gestörten Bastarden diese Koordination fehlt. Der begrenzende Faktor für die Peroxydasebildung ist bei den letzteren das Coenzym. Trotz des niedrigeren Gesamthämatingehaltes ist wesentlich mehr Hämatin in Form von Peroxydase festgelegt als bei den normalen Bastarden.Die Ergebnisse werden mit den Befunden über die Peroxydasevermehrung im alternden und im verdunkelten Blatt in Verbindung gebracht und diskutiert.Herrn Prof. Dr.W. Rudorf zum 70. Geburtstag gewidmet.     

Über die Fraktionierung des Protoplasmas vonPhaseolus-Arten undPhaseolus-Artbastarden

Zusammenfassung Es werden Beispiele für die Fraktionierung des Protoplasmas der Blätter vonPhaseolus vulgaris, Phaseolus coccineus undPhaseolus vulgaris ()×Phaseolus coccineus () mitgeteilt. Die Zahlen lassebn keine sicheren Unterschiede zwischen den entsprechenden partiklären und Grundplasmafraktionen der Elternarten und Bastarde erkennen, doch zeigen sich Differenzen, wenn die Partikel- und Grundplasmafraktionen jeweils zusammengefaßt und die Quotienten Partikel-Nucleinsäure/Grundplasma-Nucleinsäure (NS_P/NS_G) und Partikel-Eiweiß/Grundplasma-Eiweiß (E_P/E_G) gebildet werden. Diese Quotienten liegen beiPhaseolus vulgaris niedriger als beiPhaseolus coccineus, und bei den nichtgestörten Bastarden stimmt der Quotient E_P/E_G praktisch mit dem vonPhaseolus coccineus überein, wähert ist. Die gestörten Bastarde ähneln im Nucleinsäuregehalt den Elternarten und den normalen Bastarden, doch ist in allen Fraktionen, besonders stark in den Chloroplasten, der Eiweißanteil bezogen auf Frischsubstanz und Nucleinsäure vermindert.—Nucleinsäure und Eiweiß nehmen mit der Entwicklung der Blätter ab, dabei wird das Verhältnis Eiweiß/Nucleinsäure, eine Fraktion ausgenommen, stark zugungsten des Eiweißes verschoben, ferner findet eine Verschiebung der Quotienten NS_P/NS_G und E_P/E_G zugunsten der partikeln statt. Eine Abhängigkeit der Quotienten vom Gesamteiweiß oder der Gesamtnucleinsäure, die beide in den einzelnen Versuchen stark variieren, ist nicht festzustellen. Die Befunde zeigen, daß die in den Bastarden enthaltenen Kerngene vonPhaseolus coccineus das Verhältnis der Plasmakomponenten der Mutter, das bei völliger Unabhängigkeit und Selbständigkeit mit dem vonPhaseolus vulgaris übereinstimmen müßte, wesentlich beeinflussen. Sie werden unter dem Gesichtspunkt der Konkurrenzbeziehungen zwischen den Plasmakomponenten, deren Zusammenspiel den Stoffwechsel der Zellen ergibt, diskutiert.     

Movements and associations of ribosomal subunits in a secretory cell during growth inhibition by starvation

In Chironomus tentans salivary gland cells, the cytoplasm can be dissected into concentric zones situated at increasing distances from the nuclear envelope. After RNA labeling, the newly made ribosomal subunits are found in the cytoplasm mainly in the neighborhood of the nucleus with a gradient of increasing abundance towards the periphery of the cell. The gradient for the small subunit lasts for a few hours and disappears entirely after treatment with puromycin. The large subunit also forms a gradient but one which is only partially abolished by puromycin. The residual gradient which which is resistant to the addition of the drug is probably due to the binding of some large ribosomal units to the membranes of the endoplasmic reticulum (J.-E. Edstrom and u. Lonn. 1976. J. Cell Biol. 70:562-572, and U. Lonn and J.-E. Edstrom. 1976. J. Cell. Biol. 70:573-580). If growth is inhibited by starvation, only the puromycin-sensitive type gradient is observed for the large subunit, suggesting that the attachment of these newly made subunits to the endoplasmic reticulum membranes will not occur. If, on the other hand, the drug-resistant gradient is allowed to form in feeding animals, it is conserved during a subsequent starvation for longer periods than in control feeding animals. This observation provides a further support for an effect of starvation on the normal turnover of the large subunits associated with the endoplasmic reticulum. These results also indicate a considerable structural stability in the cytoplasm of these cells worth little or no gross redistribution of cytoplasmic structures over a period of at least 6 days.     

Avoidance conditioning in bamboo sharks (Chiloscyllium griseum and C. punctatum): behavioral and neuroanatomical aspects

Animals face different threats; to survive, they have to anticipate how to react or how to avoid these. It has already been shown in teleosts that selected regions in the telencephalon, i.e., the medial pallium, are involved in avoidance learning strategies. No such study exists for any chondrichthyan. In nature, an avoidance reaction may vary, ranging from a ‘freeze’ reaction to a startling response and quick escape. This study investigated whether elasmobranchs (Chiloscyllium griseum and C. punctatum) can be conditioned in an aversive classical conditioning paradigm. Upon successful conditioning, the dorsal, medial and lateral pallium were removed (group 1) and performance tested again. In a second group, the same operation was performed prior to training. While conditioning was successful in individuals of both groups, no escape responses were observed. Post-operative performance was assessed and compared between individual and groups to reveal if the neural substrates governing avoidance behavior or tasks learned in a classical conditioning paradigm are located within the telencephalon, as has been shown for teleosts such as goldfish.     

Wood decay under the microscope

Many aspects of the interactions between host wood structure and fungal activity can be revealed by high resolution light microscopy, and this technique has provided much of the information discussed here. A wide range of different types of decay can result from permutations of host species, fungal species and conditions within wood. Within this spectrum, three main types are commonly recognised: brown rot, white rot and soft rot. The present review explores parts of the range of variation that each of these encompasses and emphasizes that degradation modes appear to reflect a co-evolutionary adaptation of decay fungi to different wood species or the lignin composition within more primitive and advanced wood cell types. One objective of this review is to provide evidence that the terms brown rot, white rot and soft rot may not be obsolete, but rigid definitions for fungi that are placed into these categories may be less appropriate than thought previously. Detailed knowledge of decomposition processes does not only aid prognosis of decay development in living trees for hazard assessment but also allows the identification of wood decay fungi that can be used for biotechnology processes in the wood industry. In contrast to bacteria or commercial enzymes, hyphae can completely ramify through solid wood. In this review evidence is provided that wood decay fungi can effectively induce permeability changes in gymnospermous heartwood or can be applied to facilitate the identification of tree rings in diffuse porous wood of angiosperms. The specificity of their enzymes and the mild conditions under which degradation proceeds is partly detrimental for trees, but also make wood decay fungi potentially efficient biotechnological tools.