Polyamine als Stimulatoren von Genaktivitäten Eine autoradiographische Studie an Riesenchromosomen von Chironomus thummi

Zusammenfassung Inaktive Riesenchromosomen der Speicheldrüsen ausgewachsener Chironomus thummi-Larven zeigten nach 60 min Inkubation mit 0,05 N Putrescin-, Spermidin-oder Spermin-Lösung erneute Syntheseaktivitäten, die sich im Einbau von radioaktiv markiertem Uridin autoradiographisch zu erkennen geben. Diese Beobachtung wird im Zusammenhang mit der bekannten genaktivierenden Wirkung von Mg⁺⁺ diskutiert. Auf der Basis zahlreicher biochemisch analoger Wirkungen von Polyaminen und Mg⁺⁺ wird ein ursprünglich für die genaktivierende Wirkung von Mg⁺⁺ postuliertes Modell auf Polyamine übertragen.

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Stimulation of gene activities by polyamines an autoradiographic study with giant chromosomes of Chironomus thummi

Summary Inactive salivary gland giant chromosomes of full grown Chironomus thummi larvae showed after 60 min of incubation in 0.05 N putrescine, spermidine or spermine solutions autoradiographically demonstrable incorporation of radioactiv labelled uridine caused by a reactivation of RNA synthesis. This observations is discussed in relation to the known gene activating effect of Mg⁺⁺, and, since polyamines and Mg⁺⁺ have numerous effects in common, it is suggested that polyamines affect gene activity in a way similar to Mg⁺⁺.

     

Ueber den stand der Agrarmeteorologischen Forschung in der Deutschen Demokratischen Republik

Ohne Zusammenfassung     

Elektronenmikroskopische Untersuchungen über Glaskörperrindenzellen und Zonulafasern

Zusammenfassung Es werden folgende Befunde zur Feinstruktur des Glaskörpers beim 16 Tage alten Rattenembryo und der Glaskörperrinde beiderseits der Ora serrata beim 3 Jahre alten, an Retinoblastom erkrankten Kind mitgeteilt:Der Glaskörper des Rattenembryos und die Glaskörperrinde des kindlichen Auges enthalten Fibroblasten. Sie unterscheiden sich nicht von den im Bindegewebe vorkommenden Fibroblasten. Im embryonalen Rattenglaskörper wurden außerdem faserbildende Zellen mit wabiger Struktur des Zytoplasmas beobachtet.Die Fibroblasten der Glaskörperrinde bilden die Fibrillen des Glaskörpergerüstes und der Zonulafasern. Diese Fibrillen zeigen eine deutliche Querstreifung. Die Streifung ist unregelmäßig oder periodisch. Die Länge der Perioden beträgt in unseren Schnitten bei den Glaskörperfibrillen des Rattenembryos und des kindlichen Auges meist etwa 120, seltener 210 A. An den Fibrillenbündeln einer Zonulafaser des kindlichen Auges wurden Perioden von 90–120, 400, 440 und 630 A beobachtet.Die Fibroblasten der Glaskörperrinde des kindlichen Auges liegen im Bereich der Pars plana corporis ciliaris auch tief in den Buchten und Falten des Ciliarepithels. Hierdurch wird eine maximal große Anheftungsfläche für die von ihnen produzierten Fibrillen der Zonulafasern gewährleistet.Die Pars plana corporis ciliaris des kindlichen Auges ist von einem dichten Netz von Fibroblastenfortsätzen überzogen. Auch vereinzelte Makrophagen finden sich hier.Unsere elektronenmikroskopischen Befunde bestätigen die Angaben von Balazs über das Vorkommen von Fibrocyten (Fibroblasten) und Makrophagen in der Glaskörperrinde. Ferner bestätigen sie die bereits von früheren Autoren lichtmikroskopisch gewonnenen Erkenntnisse, wonach es sich beim Glaskörper um mesenchymales Gewebe, bei den Zonulafasern im Bindegewebsfasern handelt.

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Summary The following electron microscopical findings in the vitreous body of 16-day-old rat embryos and in the vitreoretinal border layer on both sides of the ora serrata in a 3-year-old child are reported:The vitreous body of the rat embryo and the vitreoretinal border layer of the infant eye contain fibroblasts. These fibroblasts do not differ from those present in connective tissue. The embryonic vitreous body of the rat contains fibre-forming cells, which show an alveolar structure of the cytoplasm.The fibroblasts in the cortical tissue layer of the vitreous body form the fibrils of the stroma of the vitreous body and the zonula fibres. These fibrils show a marked cross striation. The striation is either irregular or shows periodicity. In the vitreous body of the rat embryo and of the infant eye the lenght of these periods has been measured with 120 and 210 A. Periods of 90–120, 400, 440 and 630 A could be shown in the fibrillar bundles of a zonula fibre of the infant eye. In the region of the pars plana corporis ciliaris the fibroblasts of the cortical tissue layer of the vitreous body of the infant eye are also found deep down in the sinus and folds of the ciliary epithelium. Thus they guarantee an as large as possible area for the attachment of the fibrils of the zonula fibres which they produce.The pars plana corporis ciliaris of the infant eye is covered with a dense network of fibroblast processes. Also cells of the macrophage type may be found in this region.Our electron microscopical findings confirm those of earlier light microscopists and lately by Balazs. The vitreous body of the eye is formed by mesenchymal tissue, whereas the zonula fibres are formed by connective tissue.

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Mit dankenswerter Unterstützung durch den Schweizerischen Nationalfonds zur Förderung der Wissenschaftlichen Forschung.     

Conformational analysis of a toxic peptide from Trimeresurus wagleri which blocks the nicotinic acetylcholine receptor.

The 22-residue toxic peptide (WTX1) from the venom of the Southeast Asian snake Trimeresurus wagleri has multiple sites of action, but its lethal effect has been attributed to blocking the postsynaptic acetylcholine receptor at the neuromuscular junction. The 3-dimensional structure of WTX1 was studied using 2-dimensional nuclear magnetic resonance spectroscopy, circular dichroism, and computer simulations. In aqueous solution, WTX1 was shown to have extended and flexible "tails" defined by a short, rigid disulfide-bonded loop. The flexible regions can undergo structural rearrangement when moved from an aqueous to a less polar environment and may contribute to its effectiveness at different receptor sites. By substituting Gly or Phe for His at position 10, significant effects on the disulfide bond formation and, thereby, the activity of the peptide were observed. These results suggest that even subtle differences in single residues can have profound effects on the dynamics of folding, disulfide bond formation, and activity of this toxic peptide.     

Microbial oxidation of ebastine

The microbial oxidation of ebastine to carebastine was investigated. Among the 15 micro-organisms examined, only theCunninghamella strains showed the desired biotransformation.Cunninghamella blakesleeana oxidised the substrate within 7 days, via the intermediates alcohol and aldehyde, mainly to carebastine, the corresponding carboxylic acid. Optimisation of the culture conditions increased the yield from initially 10% up to a reproducible 40%. For the synthesis of carebastine a substrate concentration of 200 mg/l, a starting pH of 5.0 and the addition of 1% poly(vinyl alcohol) is favourable. The results achieved in experiments with shaking flasks are transferable to the fermentation scale and yielded 270 mg carebastine in a 3-l fermentation of 600 mg ebastine. The progress of the reaction was detected by TLC and HPLC, the products were identified by mass spectrometry and NMR.     

Xylitol production by immobilized recombinant Saccharomyces cerevisiae in a continuous packed-bed bioreactor

Continuous xylitol production with two different immobilized recombinant Saccharomyces cerevisiae strains (H475 and S641), expressing low and high xylose reductase (XR) activities, was investigated in a lab-scale packed-bed bioreactor. The effect of hydraulic residence time (HRT; 1.3-11.3 h), substrate/cosubstrate ratio (0.5 and 1), recycling ratio (0, 5, and 10), and aeration (anaerobic and oxygen limited conditions) were studied. The cells were immobilized by gel entrapment using Ca-alginate as support and the beads were treated with Al(3+) to improve their mechanical strength. Xylose was converted to xylitol using glucose as cosubstrate for regeneration of NAD(P)H required in xylitol formation and for generation of maintenance energy. The stability of the recombinant strains after 15 days of continuous operation was evaluated by XR activity and plasmid retention analyses. Under anaerobic conditions the volumetric xylitol productivity increased with decreasing HRT with both strains. With a recycling ratio of 10, volumetric productivities as high as 3.44 and 5.80 g/L . h were obtained with the low XR strain at HRT 1.3 h and with the high XR strain at HRT 2.6 h, respectively. However, the highest overall xylitol yields on xylose and on cosubstrate were reached at higher HRTs. Lowering the xylose/cosubstrate ratio from 1 to 0.5 increased the overall yield of xylitol on xylose, but the productivity and the xylitol yield on cosubstrate decreased. Under oxygen limited conditions the effect of the recycling ratio on production parameters was masked by other factors, such as an accumulation of free cells in the bioreactor and severe genetic instability of the high XR strain. Under anaerobic conditions the instability was less severe, causing a decrease in XR activity from 0.15 to 0.10 and from 3.18 to 1.49 U/mg with the low and high XR strains, respectively. At the end of the fermentation, the fraction of plasmid bearing cells in the beads was close to 100% for the low XR strain; however, it was significantly lower for the high XR strain, particularly for cells from the interior of the beads. (c) 1996 John Wiley & Sons, Inc.     

Nutrient balances and phytoplankton dynamics in two agriculturally loaded shallow lakes

The impact of agriculture was estimated on two shallow, eutrophic lakes, Lake Kotojärvi and Lake Villikkalanjärvi in southern Finland. The main emphasis was on phosphorus and nitrogen budgets and on the phytoplankton dynamics. Special attention was paid to internal P loading and blue-green algal blooms. The mean Tot-P load from agricultural land was 1.2 kg ha^-1 a^-1 in both basins and Tot-N loads were 19 kg ha^-1 a^-1 in L. Villikkalanjärvi and 12 kg ha^-1 a^-1 in L. Kotojärvi. The Tot-P input to L. Kotojärvi was on an average 0.62 g m^-2 a^-1 (per lake surface area), and the Tot-N input 9.1 g m^-2 a^-1. The corresponding inputs to L. Villikkalanjärvi were 3.1 and 57 g m^-2 a^-1, respectively. The annual variation followed the runoff volumes. About half of the Tot-P and one third of the Tot-N load was retained in L. Kotojärvi. In L. Villikkalanjärvi the retention was only 24% for Tot-P and 19% for Tot-N. The difference was very probably due to a longer theoretical retention time in L. Kotojärvi. In L. Villikkalanjärvi the mean concentration of Tot-P was 120 µg 1^-1 and that of Tot-N 1700 µg 1^-1 and the corresponding figures in L. Kotojärvi 67 and 990 µg 1^-1, respectively. The mean chlorophyll a concentration was, however, higher in L. Kotojärvi (26 µg 1^-1) than in L. Villikkalanjärvi (20 µg 1^-1). This was probably due to an internal P load in L. Kotojärvi: in 1988 the internal load of dissolved P was estimated to be as much as twofold the external load. In L. Villikkalanjärvi the internal dissolved P load was only up to 50% of the external input. In L. Kotojärvi the high internal P load coupled with a low DIN:DIP ratio resulted in a strong blue-green algal bloom in the summer of 1988. In L. Villikkalanjärvi blue-green algae were observed only in small amounts. Even in August 1990, when the DIN:DIP ratio was low enough to favor the occurrence of blue-green algae, they contributed only up to 10–15% of the total phytoplankton biomass.