Glycoconjugate pattern of membranes in the acinar cell of the rat pancreas

Summary Lectin-binding studies were performed at the ultrastructural level to characterize glycoconjugate patterns on membrane systems in pancreatic acinar cells of the rat. Five lectins reacting with different sugar moieties were applied to ultrathin frozen sections: concanavalin A (ConA): glucose, mannose; wheat-germ agglutinin (WGA): N-acetylglucosamine, sialic acid; Ricinus communis agglutinin I (RCA I): galactose; Ulex europaeus agglutinin I (UEA I): l-fucose; soybean agglutinin (SBA): N-acetylgalactosamine). Binding sites of lectins were visualized either by direct conjugation to colloidal gold or by the use of a three-step procedure involving additional immune reactions. The rough endoplasmic reticulum and the nuclear envelope of acinar cells was selectively labelled for ConA. The membranes of the Golgi apparatus bound all lectins applied with an increasing intensity proceeding from the cis-to the trans-Golgi area for SBA, UEA I and WGA. In contrast RCA I selectively labelled the trans-Golgi cisternae. The membranes of condensing vacuoles and zymogen granules were labelled for all lectins used although the density of the label differed between the lectins. In contrast the content of zymogen granules failed to bind SBA and WGA. Lysosomal bodies (membranes and content) revealed binding sites for all lectins used. The plasma membranes were heavily labelled by all lectins except for SBA which showed only a weak binding to the lateral and the apical plasma membrane. These results are in accordance to current biochemical knowledge of the successive steps in the glycosylation of membrane proteins. It could be demonstrated, that the cryo-section technique is suitable for the fine structural localisation of surface glycoconjugates of plasma membranes and internal membranes in pancreatic acinar cells using plant lectins.     

Degradation of synthetic lignins and some lignin monomers by the yeastRhodotorula glutinis

Rhodotorula glutinis degraded variously¹⁴C-labelled synthetic lignins in the presence of 0.1% glucose as co-substrate. Side chain-labelled DHP was degraded the most. While this yeastutilized vanillate and forulate for growth, sinapate/syringate were poorly or not degraded. Gallate, protocatechuate and acetate also supported growth. [carboxy-¹⁴C]Syringate was rapidly converted to¹⁴CO₂ by the yeast only in presence of glucose while [carboxy-¹⁴C]vanillate did not require any additional cosubstrate for mineralization. Ring-labelled vanillyl alcohol was also dagraded proving that the yeast could rapidly metabolize guaiacyl structures while syringyl structures required the presence of additional energy sources.

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Résumé Rhodotorula glutinis dégrade des lignines synthétiques marquées au¹⁴C de manière variée, en présence de 0.1% de glucose comme co-substrat. La DHP marquée sur la chaîne latérale est dégradée le plus. Alors que cette levure utilise le vanillate et le férulate pour sa croissance, le sinapate et le syringate sont peu ou prou dégradables. Le gallate, le protocatéchuate et l'acétate supportent également la croissance. Le syringate marqué au¹⁴C dans sa fonctioncarboxyle estrapidement convert en¹⁴CO₂ par le levure mais exclusivement en présence de glucose, tandis que le vanillate marqué au¹⁴C dans sa fonction carboxyle ne requiert aucun co-substrat additionel pour sa minéralisation. L'alcool vanillique marqué dans son cycle, est également dégradé démonstrant ainsi que la levure peut métaboliser rapidement des structures guaiacyliques tandis que les structures syringiques requièrent la présence de sources auxiliaires d'énergie.

     

Plasmid content and homology of 16 strains of filamentous,nonheterocystous cyanobacteria

We have developed several strain-specific, rapid, small-scale plasmid isolation procedures in order to characterize the plasmid profiles of 16 filamentous, nonheterocstous cyanobacteria. At least one distinct plasmid was found in eight strains, with seven of these containing two or more different plasmids. Eight strains were found to be without plasmid DNA. Both the large, 12.9 kb, and the small, 1.6 kb, plasmids fromPlectonema boryanum 581 were isolated, purified, and cloned. Southern blots of plasmid DNAs from the eight strains were probed with these cloned DNAs and also with ultra-pure plasmid DNA fromPhormidium liridum 426. Four strains ofP. boryanum (485, 581, 594, 1542) andP. luridum 426 have identical plasmid profiles, and plasmid homology is extensive.     

Promoters active in mammalian cells (pSV40) and in plants (pNOS) permit expression in yeast of the Tn5 APH(3') II gene

Abstract Four plasmids were constructed by associating Escherichia coli and yeast selection markers and replication origins to a structural gene coding for aminoglycoside phosphotransferase (APH(3')) controlled by different flanking sequences. We used the two bacterial genes of Tn5 (APH(3')II) and Tn903 (APH(3')I) as such and the chimeric pSVneo (APH(3')II) and pNOSneo (APH(3')II) constructs, functional in mammalian and plant cells, respectively. Yeast clones resistant to G418 were obtained with all plasmids except with that bearing the bacterial APH(3')II gene. The three plasmids harbouring the functional APH genes, however, conferred different levels of G418 resistance to yeast.     

Growth,aluminium uptake and mucous cell morphometrics of early life stages of brown trout,Salmo trutta,in low pH water

Synopsis Young (7–10 days after hatching) brown trout (Salmo trutta) exposed for 5 days to pH 5 in high calcium water and at 2 temperatures (12°, 4°C) in the laboratory displayed no alterations in growth or in mucous cell concentration and volume, compared to the control group kept at pH 7.2. Contamination of acid-stressed young with 230 μg All^-1 resulted in significant growth depression and Al accumulation, but in no changes of mucous cell morphometrics. Field tests in low calcium water produced high mortality at low pH (5.1), but showed consistent effects on mucous cells as in laboratory experiments. Three-month-old juveniles of brown trout, subjected to decreased pH values at 12° and in high calcium water for 8 days exhibited mucous cell hyperplasia (without hypertrophy) within 3 h of the acid addition. After 120 h sloughing of the integument occurred with full recovery not possible within a 4-day-recovery period. Although the results presently apply only to hard water conditions, the differences between juveniles and recently hatched young in tolerance to pH- and Al-mediated stress may also be of importance for soft waters affected by acid rain.     

Structure and function of ferredoxin-NADP+-oxidoreductase

The redox-enzyme ferredoxin-NADP-oxidoreductase has been shown to be activated by light and inactivated in the dark. This review will summarize recent data concerning the biochemical characterization of the enzyme compared to its in-vivo activation. Further-more the mechanism of this activation process is discussed as a conformational change caused by the light-driven proton gradient.Abbreviations cyt cytochrome - fd ferredoxin - FNR1 large form of ferredoxin-NADP-oxidoreductase - FNRox oxidized FNR - FNRred reduced FNR - FNRs small form of FNR - FNRsq FNR-semiquinone     

Comparison of two lignin-degrading fungi:Phlebia radiata andPhanerochaete chrysosporium

Summary The ligninolytic enzymes ofPhlebia radiata were produced in static conditions earlier developed forPhanerochaete chrysosporium. The production pattern of lignin peroxidases resembled that ofP. chrysosporium. The extracellular proteins ofPhlebia radiata were separated by isoelectric focusing. Four proteins with acidic isoelectric points (4.15) were detected by peroxidase staining. The peroxidases ofP. radiata reacted with antibodies produced against a peroxidase ofPhanerochaete chrysosporium and vice versa. Thus the lignin peroxidases of the two fungi have major similarities despite slight differences in their isoelectric points and molecular weights. Veratryl alcohol was produced by both fungi and degraded to veratraldehyde, two lactones and a quinone by the ligninolytic cultures.     

Energy metabolism,body-temperature and breathing parameters in nontorpid blue-naped mousebirdsUrocolius macrourus

Summary Breathing frequencyF _r of resting blue-naped mousebirdsUrocolius macrourus lies between 50–70 per min and correlates directly with ambient temperatureT _a and energy metabolismM. The nocturnal mean energy intake per breath varies between 5.6–17.7 mJ/g. At highT _a the birds show gular fluttering with a relatively constantF _r of about 460 min^–1.M shows a constant absolute day-night difference of 25 J/g·h; the relative differences areT _a-dependent between 36–168% (lower values at lowerT _a). Thermal conductance is 2.10–2.15 J/g·h·°C (predicted 2.67), indicating a good insulation. Basal metabolic rate BMR is reduced by 63% compared to predicted values. At aT _a-range of +8–36 °C the birds are normothermic. Below this range nocturnalT _b andM decrease slightly with fallingT _a. The birds show partial heterothermia (shallow hypothermia). Clustering is an effective energy saving strategy which allows loweringM with keeping highT _b even at lowT _a.Oxygen-intake is controlled byF _r as well as by tidal volumeV t inT _a-dependent changing portions.V T can vary between 0.29–0.91 ml (mean value 49.7 ml).Abbreviations T _a ambient temperature - T _b body temperature - M energy metabolism - F _r breathing frequency - V T tidal volume - BMR basal metabolic rate - TNP thermoneutral point     

Scopolin, a Biochemical Marker for Resistance to Thielaviopsis basicola in Callus and Crown-Gall Tissue Cultures of Tobacco

Scopolin concentration increased in tissue cultures where Thielaviopsis basicola growth ceased (primary and established callus cultures of the resistant tobacco cultivar Ky 170) while it decreased in tissue cultures where fungal growth persisted (primary and established callus cultures of the susceptible tobacco cultivar Ky 151 and crown-gall cultures of Ky 170 and Ky 151). The concentration of chlorogenic acid, scopoletin, and two other unknown soluble phenols varied after inoculation and no, correlation with tissue culture resistance could be established. Incorporation of Agrobacterium tumefaciens T-DNA in the genome of both cultivars induced a drastic reduction of scopolin in inoculated tissue cultures. T-DNA incorporation had less, influence on uninoculated tissues. Scopolin at concentrations found in tissue cultures was not toxic to the fungus.     

Resistance of Tobacco to Thielaviopsis basicola under Tissue Culture Conditions and Increased Susceptibility after Transformation with Agrobacterium tumefaciens T-DNA

Tobacco callus cultures from a Thielaviopsis basicola (Berk. et Br.) Ferraris resistant cultivar were less severely colonized than callus cultures from susceptible cultivars by the pathogen at all concentrations of kinetin and α-indoleacetic acid tested. However, at concentrations where these substances influenced the morphology of the callus cultures they also influenced the degree of colonization by the fungus. Incorporation of T-DNA of Agrobacterium tumefaciens induced susceptibility in tissue culture of the resistant cultivar but did not significantly modify the reaction of the susceptible cultivar tested.