Comparison of fatty acids of marine fungi using multivariate statistical analysis

Summary Ten obligate marine fungi have as their principal fatty acids 160, 180, 181n9 and 182n6. The fatty acids ranged from 14 to 22 carbons, completely dominated by those with even numbers of carbons. The amount of unsaturated fatty acids varied between 35% and 80%. Each isolate contained small amounts of the acids 183n3 and 204n6. Branched, hydroxy- or cyclic fatty acids were not detected. Multivariate statistical, i.e. principal component analysis, showed that all ten strains could be distinguished on the basis of their fatty acid composition. These results indicate that the marine fungi do not have an unusual fatty acid composition and suggest that chemometric, multivariate analysis might be employed to confirm taxonomic relationships among these organisms.     

Fehlender Einfluß von Licht auf die Reversion der UV-induzierten spektralen Veränderung von Cytidylsäure

Zusammenfassung Das Lichtprodukt, welches nach Belichtung von UV-bestrahlter Cytidylsäure (CMP) in früheren Versuchen zu entstehen schien und über das von Park u. Kaplan berichtet worden war, konnte in neuen Versuchen des zweiten Autors nicht wiedergefunden werden. Nachbehandlung mit Licht wie auch Dunkelheit bei ca. 25°C ergaben offenbar die gleiche Substanz mit dem Absorptionsspektrum von normalem CMP.

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Summary The photoproduct after illumination of UV-irradiated Cytidylic acid (CMP) reported by Park and Kaplan earlier could not be reproduced by the 2^nd author. Both, postincubation with and without light at about 25°C led to the formation of obviously the same substance with the absorption spectrum of normal CMP.

     

Selectivity of ATP-activated GTP-dependent Ca2+-permeable channels in rat macrophage plasma membrane

Outside-out configuration of the patch clamp technique was used to test whether an intracellular application of G protein activator (GTPS) affects ATP-activated Ca²⁺-permeable channels in rat macrophages without any agonist in the bath solution. With 145 mm K⁺ (pCa 8.0) in the pipette solution, activity of channels permeable to a variety of divalent cations and Na⁺ was observed and general channel characteristics were found to be identical to those of ATP-activated ones. Absence of extracellular ATP makes it possible to avoid the influence of ATP receptor desensitization and to study the channel selectivity using a number of divalent cations (105 mm) and Na⁺ (145 mm) as the charge carriers. Permeability sequence estimated by extrapolated reversal potential measurements was: Ca²⁺ Ba²⁺ Mn²⁺ Sr²⁺ Na⁺ K⁺ = 68 30 26 10 3.5 1. Slope conductances (in pS) for permeant ions rank as follows: Ca²⁺ Sr²⁺ Na⁺ Mn²⁺ Ba²⁺ = 19 18 14 12 10. Unitary Ca²⁺ currents display a tendency to saturate with the Ca²⁺ concentration increase with apparent dissociation constant (K _d ) of 10 mm. No block of Na⁺ permeation by extracellular Ca²⁺ in millimolar range was found. The data obtained suggest that (i) activation of some G protein is sufficient to gate the channels without the ATP receptor being occupied, (ii) the ATP receptor activation results in the gating of a special channel with the properties that differ markedly from those of the receptoroperated or voltage-gated Ca²⁺-permeable channels on the other cell types.DeceasedThe authors are grateful to K. Kiselyov and A. Mamin for technical assistance. The work was supported by the Russian Basic Research Foundation, Grant N 93-04-21722 and was made possible in part by Grant N R4A000 from the International Science Foundation.     

Über den Einfluß des Salzgehaltes auf die photosynthetische Leistung verschiedener Standortformen vonDelesseria sanguinea undFucus serratus

Zusammenfassung 1. Es wurde untersucht, welchen Einfluß kurzfristige und langfristige Salzgehaltsveränderungen auf verschiedene Standortformen der RotalgeDelesseria sanguinea und der BraunalgeFucus serratus haben. Als Kriterium des Lebenszustandes wurde die photosynthetische Leistung gewählt. Die Algen wurden folgenden Salzgehaltskonzentrationen ausgesetzt: 0, 5, 10, 15, 20, 30, 40, 50 S.2. Die Versuche ergaben, daß kurzfristige Konzentrationsveränderungen (30 min) — sowohl Erniedrigung als auch Erhöhung des Salzgehaltes — die photosynthetische Leistung stimulieren. Ein langfristiger Aufenthalt (24 Std) unter den veränderten Bedingungen bewirkt, sofern diese innerhalb der Toleranzgrenzen der Algen liegen, einen Ausgleich der anfänglichen Stimulation. Außerhalb der Toleranzgrenzen liegende Konzentrationen rufen nach der Stimulation eine Leistungsdepression hervor. Bei Rückübertragung in den Ausgangssalzgehalt sind die Depressionen teilweise reversibel.3. Im hypotonischen Milieu verhalten sich die Delesserien der verschiedenen Standorte (Helgoland, Kattegat, Kieler Bucht) gleich: in 5 S treten starke Depressionen auf. Nordsee-Delesserien sind im hypertonischen Milieu weniger empfindlich, sie zeigen noch bei 50 S eine gesteigerte photosynthetische Leistung. In diesem Bereich sind die Ostseeformen schon schwer geschädigt. Am empfindlichsten gegenüber allen Konzentrationsänderungen ist die BrackwasserformDelesseria sanguinea formalanceolata aus der Kieler Bucht.4.Fucus serratus aus dem Litoral von Helgoland zeichnet sich im Gegensatz zu der submers lebenden Form der Ostsee, die sich ähnlich wieDelesseria verhält, in allen untersuchten Konzentrationsbereichen durch eine unveränderte photosynthetische Leistung aus. Die beiden Standortformen vonFucus entsprechen gemäß der Einteilung vonMontfort (1931) dem resistenten Typ und dem Stimulations-Depressionstyp.

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On the influence of salinity on photosynthetic performance of various ecotypes ofDelesseria sanguinea andFucus serratus

The phaeophyceanF. serratus and the rhodophyceanD. sanguinea came from the North Sea (30 S) and the Baltic Sea (15 S). The activity of photosynthesis was taken as a criterion of algae vitality. Experiments were made in salinity concentrations of 0, 5, 10, 15, 20, 30, 40 and 50 S. Thirty-minute exposures to sub- or supranormal salinities stimulate photosynthesis. Within their physiological salinity ranges the algae assume normal photosynthetic rates within 24 hours. Extreme salinities cause a reduction in photosynthetic activity; this reduction mostly disappears, however, after re-transfer into normal salinity conditions. At 5 S all test individuals ofDelesseria from different locations exhibit a reduction of photosynthetic rates. At 50 SDelesseria from the North Sea still show increased activity, whileDelesseria from the Baltic are already severely damaged. The brackish-water formD. sanguinea (formalanceolata) is most sensitive to salinity variations. The photosynthetic activity ofF. serratus from Helgoland does not vary in all salinities employed. The range of test salinities corresponds to that of the habitat in the littoral zone, where high salinities occur during air exposure, and low salinities, during rainfall. By contrast, inF. serratus from the Baltic Sea occurring only in the sublittoral zone, photosynthetic rates are similarly affected by salinity as inDelesseria.

     

Hat J. G. Mendel bei seinen Versuchen „zu genau“ gearbeitet? — Der χ2-Test und seine Bedeutung für die Beurteilung genetischer Spaltungsverhältnisse

Zusammenfassung R. A. Fisher (1936) hat im Zusammenhang mit einer statistischen Analyse der Versuche GregorMendels die Auffassung geäußert,Mendels Versuchsergebnisse seien statistisch gesehen zu exakt. DaMendel vielleicht auf Grund der ersten Spaltungen die obwaltende Gesetzmäßigkeit bereits erkannt habe, könne den weiteren Versuchen wohl nur demonstrativer Wert zuerkannt werden. Andere Autoren, z.B. Zirkle (1964) sowiede Beer (1964), schließen sich diesem Urteil an. Fisher — und mit ihm die genannten Autoren — haben jedoch übersehen, daß bei Beurteilung der F₃-Analysen derMendelschen Versuche, soweit nicht Samen-, sondern Pflanzenmerkmale geprüft wurden, die die Aufzucht einer F₃ erforderlich machten, die Größe der ausgewerteten Nachkommenschaften kaum gleich der Zahl der jeweils ausgelegten oder als ausgelegt angenommenen Samen 10 gewesen sein kann, da mit Auflaufschäden, Verlust durch Vogelfraß usw. gerechnet werden muß. Unsere Untersuchung zeigt, daß bei Annahme einer effektiven Größe dieser Nachkommenschaften von durchschnittlich 8 Pflanzen sich zahlenmäßig fast die gleiche Gesamtwahrscheinlichkeit für die Übereinstimmung aller Versuche mit der Erwartung ergibt, die sich als Gesamtwahrscheinlichkeit auch für die Versuche an Erbsen beiCorrens (1900),Tschermak (1900),Bateson undKillby (1905) sowieDarbishire (1908, 1909) — diese zusammengerechnet — ermitteln läßt. Überdies steht ein Ausfall von 10–20% des ausgelegten Erbsensaatgutes sowohl mit den AngabenMendels wie den Erfahrungen mährischer Pflanzenzüchter bei modernen Erbsensorten in Einklang bzw. ist leicht in Einklang zu bringen.Die scheinbar übergroße Exaktheit der Erbsenversuche ist wohl darauf zurückzuführen, daß die genetischen Spaltungszahlen, offensichtlich je nach Pflanzen- oder Tierart verschieden, nicht binomial, sondern halb zufällig verteilt sind und aus diesem Grunde die errechnete Größe ² zu klein ausfällt.Es wird versucht, den Faktorc zu schätzen, um den der ²-Wert bei der Erbse zu klein ausfällt, und die Konsequenzen zu ermitteln, die diese Verhältnisse für die Gültigkeit des ²-Testes besitzen. Zum Vegleich werden die Ergebnisse mitgeteilt, die eine ähnliche Untersuchung bei mehreren Pflanzen- und Tiergattungen ergab, wobei ebentalls Daten aus der Literatur zugrunde gelegt wurden.

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Has J. G. Mendel been too accurate in his experiments? The ² test and its significance to the evaluation of genetic segregation

Summary In a statistical analysis ofGregor Mendel's experimentsR. A. Fisher (1936) expresses the opinion that from a statistical point of viewMendel's experimental results are too exact. Assuming thatMendel recognized the regularities of segregation already from his first seed counts in 1858, Fisher believes that further experimentation byMendel was only of demonstrative value. Several authors, f.i. C. Zirkle (1964) share this opinion.However,Fisher and the other authors have overlooked that in judging F₃ analyses ofMendel's experiments, when not seed characteristics were tested but plant traits that made raising an F₃ necessary, the number of progeny available for classification could hardly be equal to the 10 seeds planted or presumably planted, since one has to count on losses through poor germination, birds, or other causes. We show that with an assumed average number of 8 plants in these progenies the probability of agreement with expectation in all ofMendel's experiments is numerically equal to the probability calculated from experiments with peas byCorrens (1900),Tschermak (1900),Bateson andKillby (1905), as well as byDarbishire (1908, 1909), the latter totaled.The too great seeming exactness of the experiments with peas could be explained in the following manner: The distribution of genetic segregation data, obviously different for each plant or animal species, is not binomial but semirandom, therefore the calculated ² value will be too small.We try to estimate the factor c by which the ² value in experiments on peas is too small, and to determine the consequences of this fact to the validity of the ² test. For comparison we point out results from similar investigations on several plant and animal species, again using data from the literature.

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Nach Vorträgen auf dem 13. Biometrischen Colloquium der Deutschen Region der Internationalen Biometrischen Gesellschaft in Mainz (31. 3.–2. 4. 1966) und der II. Internationalen Berliner Tagung über Mathematische Statistik und ihre Anwendungen (Berlin 9.–13. 5. 1966).     

Betrachtung über den Energiekonsum von Tieren mit Atmungsorganen von zweierlei Typ

Zusammenfassung Besitzt eine isometrisch wachsende Tierart zweierlei Atmungsorgane, von denen das eine O₂ prop. L ² aufnimmt und das andere prop. L ³, so ergibt sich in einem Koordinatensystem mit log L als Abszisse und log (O₂) als Ordinate für die Abhängigkeit des (O₂) von L keine Gerade, sondern eine gegen die Ordinate konkave, Linie (Abb. 1), die bei L=O mit der Steigung a=tg =2 beginnt, für L=1 den Wert 2,5 erreicht und bei L 8 mit der Steigung 3 endet. (Bei log G als Abszisse betragen die entsprechenden Werte 2/3, 5/6 und 1.) Für Längenintervalle von einer Zehnerpotenz (also Gewichtsintervallen von 10³) dürfte im Rahmen der experimentellen Breiten oft eine anscheinend lineare Abhängigkeit gefunden werden. Ergibt sich dann empirisch für ein Längenintervall l10 (mit der mittleren Länge L _m) oder das entsprechende Gewichtsintervall l10³ eine Atmungsgerade mit der Steigung , so kann unter den obigen Voraussetzungen vermutet werden, daß sich die durch beide Organe konsumierten O₂-Mengen verhalten wie (3 — )( — 2). Bei der Körperlänge L * (Formel 9), die nicht realisiert zu sein braucht, müßten beide Organe gleichviel O₂ aufnehmen. Die Anwendung dieser Folgerungen auf Tiere, die möglicherweise Atmungsorgane von mehrererlei Typ besitzen (Insekten, Insektenlarven, Isopoden, Schnecken), sowie die Erörterung solcher empirischer Atmungsgeraden, die eine zwischen 2 und 3 liegende Steigung a aufweisen, wird in den weiteren Mitteilungen erfolgen.Zugleich Mitt. IX der Reihe Körpergröße, Körperzeiten und Energiebilanz in dieser Zeitschrift: I: Ludwig, 24,319 (1937); II: Kittel 28, 533 (1941); VIII: Hempel, 36,261 (1954).Erarbeitet an der obengenannten, von Prof. J. M. Pérès geleiteten Station an Schlechtwettertagen, die andere Untersuchungen unmöglich machten. Ihm und der Deutschen Forschungsgemeinschaft bin ich zu großem Danke verpflichtet.     

Toxigenic fungi and the deterioration of Nigerian poultry feeds

Growth of toxigenic strains of Aspergillus clavatus Des. and Aspergillus flavus Link at 30°C on milled poultry feeds led to a considerable decrease in the protein, oil and crude fibre contents of the feed substrate. A corresponding increase in the free fatty acid fractions of the feeds due to the activities of these microbes was also recorded. Rapid degradation of the feedstuff by both species was recorded at a temperature of 25°C and 30°C and a pH range of 4.8–6.4. When grown on feed infusion broth at 30°C, the highest amounts of mycelial production with sporulation of both fungal species occurred within the 8-day incubation period. A determination of their extra-cellular enzyme profile showed the production of amylases, pectate lyase, cellulases, proteases, lipases, xyalanases, DNase and RNase.All the carbon and nitrogen sources used (except L-sorbose and DL-tryptophan), supported good mycelial growth with sporulation. An optimal CN ratio of 5.04.5 and 7.53.0 was recorded for growth and sporulation of A. clavatus. For A. flavus, a CN ratio of 7.54.5 was found best for growth and 5.03.0 for sporulation.     

Free Ia Eα chain expression in the E α + :E β − : recombinant strain A.TFR5

Molecular and biochemical techniques have been used to explore the reasons behind low E chain expression in the E ⁺ E ^– I-region recombinant strain, A.TFR5. A.TFR5 (A ^f E ^k, ap5), a recombinant between A.CA (A ^f E ^f) and A.TL (A ^k E ^k), carries the E ^k subregion. Previous results have shown that it expresses the E chain, but at reduced levels relative to E ⁺ E ⁺ strains. No E chains were detected, which is consistent with the A.TFR5E gene being derived from the A.CA parent, which carries the null E ^f allele. In this paper, the defect in E-chain expression is explored. Restriction fragment length polymorphism analysis has localized the recombination event in A.TFR5 approximately 30 kb upstream of E, in the region of the large intervening sequence of E. Northern blot analysis of total RNA from A.TFR5 shows normal amounts of the E message, but no E message. Two-dimensional gel analysis of 15 min pulse-labeled A.TFR5, A.CA, and A.TL E immunoprecipitates shows decreased levels of the intracellular E chain in A.TFR5 relative to A.TL. However, analysis of total cell extracts shows normal levels of this protein. A glycoprotein fraction isolated from total cell extracts of 5 h labeled cells contains normal amounts of intracellular E, but decreased amounts of the mature cell-surface protein. These data suggest that in the absence of E, the E chain (1) takes on an altered conformation that is not as efficiently recognized by alloantibodies, and (2) is found in normal levels as the partially glycosylated intracellular precursor, but is not processed and/or transported efficiently to the cell surface.     

Das Fettsäurespektrum des Flußkrebses Orconectes limosus und seine Beeinflussung durch die Nahrung

Zusammenfassung Das Fettsäurespektrum von männlichen Flußkrebsen Orconectes limosus wurde gaschromatographisch analysiert. Die Fettsäuren zeigen eine spezifische Verteilung auf die Lipidklassen. Das Gesamtspektrum entspricht dem Schema eines Süßwassertieres im Winter mit den größten Fraktionen: C160, C181, C202 und C204 (zusammen rund 53% der Gesamtfettsäuren). In verschiedenen Fraktionen wurden ungeradzahlige und verzweigte Fettsäuren gefunden.Durch Verfüttern von Kartoffeln und Lebertran kann das Spektrum beeinflußt werden. Die Fettsäuren in der Mitteldarmdrüse, und in weniger starkem Ausmaß im Restkörper, haben sich nach viermonatiger Fütterung mit Lebertran qualitativ und quantitativ dessen Spektrum angepaßt. Nach siebenmonatiger Fütterung mit Kartoffeln zeigen Mitteldarmdrüse und Restkörper ein Spektrum, das dem eines unbehandelten Kontrollkrebses ähnelt und sowohl C182 als auch höher ungesättigte Fettsäuren enthält. Hierfür kommen zwei Deutungen in Frage: Entweder wird das Spektrum der Fettsäuren in Strukturlipiden aufrechterhalten, oder es erfolgt de novo Synthese einer C18:2^6,9-Fettsäure und deren Verlängerung analog dem von Wirbeltieren bekannten Weg.

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The fatty acid composition in the crayfish, orconectes limosus, and the effect of nutrition

Summary The fatty acid composition in the male crayfish, Orconectes limosus, was analysed by gas-liquid chromatography. The fatty acids were found to have a specific distribution in different lipid classes. The composition corresponds to those of a fresh water animal in winter with the most important acids: C160, C181, C202, C204 (including about 53% of the total acids). In several lipid classes oddnumbered and branched chain fatty acids could be detected.Potatoe and fish-liver oil diets influence the fatty acid composition. After four months feeding with fish-liver oil the fatty acids in the hepatopancreas and to a lesser extent in the rest body show a similar spectrum as the fed oil. After feeding for seven months with potatoes hepatopancreas and rest body exhibit a fatty acid composition representative for the untreated animal, which contains C182 as well as higher unsaturated acids. These findings support the hypotheses that either the fatty acid composition in the structure lipids was maintained or that a C182^6,9-fatty acid was de novo synthesized and elongated analogous to the known vertebrate pathway.

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Herrn Prof. Dr. K. Urich bin ich für die Anregung zu der vorliegenden Arbeit und für die Durchsicht des Manuskriptes zu großem Dank verpflichtet.     

A novel 2-oxoglutarate-dependent dioxygenase involved in vindoline biosynthesis: characterization,purification and kinetic properties

The enzyme, desacetoxyvindoline 4-hydroxylase, was purified to apparent homogeneity from Catharanthus roseus by ammonium sulfate precipitation and successive chromatography on Sephadex G-100, green 19-agarose, hydroxylapatite, -kg sepharose and Mono Q. The 4-hydroxylase was characterized by its strict specificity for position 4 of desacetoxyvindoline suggesting it to catalyze the second to last step in vindoline biosynthesis. The molecular mass of the native and denatured 4-hydroxylase was 45 kDa and 44.7 kDa, respectively, suggesting that the native enzyme is a monomer. Two-dimensional isoelectric focusing under denaturing conditions resolved the purified 4-hydroxylase into three charge isoforms of pIs 4.6, 4.7 and 4.8. The purified 4-hydroxylase exhibited no requirement for divalent cations, but inactive enzyme was reactivated in a time-dependent manner by incubation with ferrous ions. The enzyme was not inhibited by EDTA or SH-group reagents at concentrations up to 10 mM. The mechanism of action of desacetoxyvindoline 4-hydroxylase was investigated. The results of substrate interaction kinetics and product inhibition studies suggest an Ordered Ter Ter mechanism where -kg is the first substrate to bind followed by the binding of O₂ and desacetoxyvindoline. Their K _m values for -kg, O₂ and desacetoxyvindoline are 45 M, 45 M and 0.03 M, respectively. The first product to be released was deacetylvindoline followed by CO₂ and succinate, respectively.Abbreviations -kg -ketoglutarate or 2-oxoglutarate - NMT N-methyltransferase - SAM S-adenosyl-l-methionine - TLC thin layer chromatography - VBL vinblastine - VCR vincristine     

Style morph frequencies in Minnesota populations of Lythrum (Lythraceae)

Style morph frequencies (shortmidlong) were determined for a total of n = 11 918 plants in 16 Minnesota populations of Lythrum salicaria L. Nine populations were in the establishment phase, with population sizes ranging from n = 56 to n = 2 192. Most of these populations exceeded previously reported population sizes in the native European habitat. A nonparametric statistical test, the chi-square (²), can be used to determine if populations are at isoplethic equilibrium (111, shortmidlong); a ² value >5.99 is significant at the 5% level. Only one established population (White Bear Lake, n = 1991, ² = 3.0) fitted the null hypothesis for isoplethy, although all established populations contained all three style morphs. Pooled values for these populations indicated an excess of mids and longs, with shorts being deficient. Colonizing populations had a higher percentage of mids (54%) when compared to established populations (33.7%). Short styles were almost nonexistent (8%) in colonizing populations. Five out of the seven populations lacked at least one style morph. A review of the literature reporting style morph frequencies in tristylous L. salicaria revealed that no statistical analysis for isoplethy has been performed. Darwin originally assumed that all populations would be isoplethic, possessing equal numbers of all three style morphs, but concluded, without statistical analysis, that, instead, populations were anisoplethic. Since tests for statistical deviations from the expected frequencies (111) have not been used, ² analysis was performed. Several of these populations were at isoplethic equilibrium (Nadder su2 = 1.7, Blelham ² = 1.69, Potsdam ² = 1.5, Vestfold ² = 0.4, Buskerud ² = 5.62, Kilchberg ² = 0.35, Lausanne ² = 3.32, Canberra ² = 5.29, Massachusetts ² = 3.13), suggesting that the general conclusion of anisoplethy in tristylous L. salicaria is inappropriate.This is Scientific Journal Series Paper Number 19 128 of the Minnesota Agricultural Experiment Station     

Structure and properties of aCephalosporium acremonium α-galactosidase

The amino acid and sugar composition of the enzyme protein, the effect of urea, sodium dodecyl sulphate and Concanavalin A on the purified -galactosidase (EC 3.2.1.22) from the moldCephalosporium acremonium has been studied. The results obtained by gas liquid chromatography indicated the presence ofN-acetylglucosamine, mannose, galactose andN-acetylneuramic acid in the molar proportions 27311. The presence of two types of Asn-linked oligosaccharide structures in the enzyme molecule is assumed. The -galactosidase liberates (1–3), (1–4) and (1–6)-linkedd-galactose units from various synthetic and natural substrates which have been tested. The effects of pH, substrate concentration and temperature on the catalytic activity of the enzyme are described. The purified -galactosidase also exhibited a lectin activity with an affinity towards glucose, and to some extent mannose.Abbreviations p-NPG p-nitrophenyl--d-galactopyranoside - 4-MUG 4-methylumbelliferyl--d-galactopyranoside - HU hemagglutinin unit - PBS phosphate buffered saline - SDS sodium dodecyl sulphate - ConA Concanavalin A - WGA wheat germ agglutinin - LCA Lens culinaris agglutinin - PHA phytohemagglutinin fromPhaseolus vulgaris     

Influence of photoperiod,temperature and host plant on the production of diapause eggs inPetrobia (Tetranychina) harti (Acari: Tetranychidae)

Petrobia harti (Ewing) diapauses in the egg stage. Adult females lay either diapause or nondiapause eggs. On the University of Thessaloniki campus (41°N), the mite was found to develop on leaves ofOxalis corniculata L. throughout the year, while no mites were found on leaves ofOxalis articulata Savigny growing in the same area. In the laboratory the mite could be maintained equally well on detached leaves of both plant species, kept on wet cotton-wool.Forty to 90% females laying diapause eggs (dlf) were produced when the mites developed under LD 1212 and 19±1 °C, or LD 168 and 19±1 °C or 25±1 °C on leaves ofO. articulata detached from plants grown in the open in various seasons. Under the same conditions, a very low to zero percentage ofdlf was produced onO. corniculata. By rearing certain feeding stages on one of these twoOxalis hosts, and the other feeding stages on the other host, various percentages ofdlf were obtained. These percentages were the net effect of the antagonistic action of the twoOxalis species.By rearing the mites at LD 8.515.5, LD 1212 or LD 168 and a temperature of 19±1 °C onO. articulata leaves renewed every 3 days, or every 16–18 days, or not at all, it could be shown that diapause induction or aversion is caused by the direct effect of photoperiod on the mites, and not by an effect through the host leaves.When wholeO. articulata plants were grown under LD 168 and 19±1 °C in the laboratory, or developed in the open during April and May, flowers were produced, while under LD 1212 no flowering occurred. In the laboratory under diapause-inducing conditions, higher percentages ofdlf were produced on leaves detached from flowering plants than on leaves detached from plants not flowering.OnO. articulata leaves at 20 °C, photoperiods with photophases equal to or longer than 12 h induced from 70 to 80%dlf, while photoperiods with photophases equal to or shorter than 10.9 h induced very low to zero percentages. By transferring different chrysalis stages from a diapause-inducing (LD 1212) to a diapause-averting (LD 8.515.5) photoperiod, and vice versa, it was found that the nymphochrysalis through deutonymph stages were sensitive to photoperiod, the deutochrysalis and deutonymph being the most sensitive.Under an LD 1212 photoperiod, a temperature of 20 °C induced diapause, whereas 25 °C, 30 °C, or a daynight thermoperiod of 25 °C18 °C suppressed it.     

Beobachtungen in vitro an den Zellen der Larvalen Ovariolenhülle von Bombyx mori L. (Lepidoptera) in Verschiedenen Kulturmedien

Zusammenfassung Die Hüllzellen der Ovariolen von Bombyx-Altlarven sind geeignet für Zellkulturen im hängenden Tropfen. Die Erfahrungen, welche mit Kulturmedien verschiedener Autoren und mit einigen Varianten gesammelt worden sind, werden mitgeteilt.Unter günstigen Bedingungen wandern die Hüllzellen vom explantierten Ovariolenstück ab und bevölkern oligogonal amöboid die Umgebung. Mitosen, aber auch zweikernige Zellen nach Amitose treten auf. Inaktive polygonale Zellen bilden ein Netzwerk oder ein Pseudoepithel. Alternde Zellen speichern Stoffwechselprodukte; ihre Kerne degenerieren. Die Kultur stirbt nach etwa 1 Woche und spätestens nach 3 Wochen, auch wenn das Mutterstück entfernt und das Medium erneuert worden ist.Unter ungünstigen Bedingungen wandern die Zellen nicht ab und verfärben sich, oder es treten fadenförmige Hungerzellen oder stark vakuolisierte Mastzellen auf.Da sich in situ nicht verschiedene Funktionszellen differenzieren, haben die kultivierten Zellen wahrscheinlich den gleichen entwicklungsphysiologischen Anfangszustand. Ein Inventar ihrer Form- und Strukturmerkmale in vitro wird aufgestellt. Einige werden als Kriterien für die Güte des Mediums ausgewählt.Die Ergebnisse in 664 Explantaten in nährstoffarmen und -reichen Kulturlösungen ohne und mit Zusatz von Hämolymphe oder Eiextrakt werden beschrieben (Tabelle, S. 406).Salzlösungen mit Zucker reichen nicht aus, dagegen solche mit Zusatz von Aminosäuren, wie sie in der Raupenhämolymphe vorkommen (Wyatt), oder von Eiweißhydrolysat (Vago). Voraussetzung ist ein molares Verhältnis Na K von etwa 0,2 1. Der Zusatz von Hämolympheextrakt bringt auch für ungünstige Kulturlösungen eine Verbesserung, der von Eiextrakt führt zur Hypertrophie. Wirbeltierextrakte haben sich nicht bewährt. Trehalose hat keinen Vorteil gebracht.Die Erfahrungen werden im Hinblick auf die Angaben anderer Autoren diskutiert. Wesentlich für die Kultur von Insektenzellen sind: das art- und stadienspezifische molare Verhältnis der Na K Kationen, die Anwesenheit von Zucker und Proteinderivaten und von Spurenstoffen, die vorläufig nur mit Extrakten aus dem Spender-Organismus gegeben werden können.

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Summary The cells of the intermediate layer enveloping the ovarioles of the old Bombyx larvae may be cultured by the hanging drop method. Using various culture fluids (methods see p. 395; modifications see p. 405) the following results are obtained: Under favourable conditions the intermediate cells will leave the explanted piece of ovariole by ameboid movement and populate the surrounding area. They assume oligogonal shape. Normal mitosis occurs but binucleate cells are also formed by amitosis. Inactive polygonal cells form either a network or a pseudoepithelium. Aging cells accumulate metabolic products; their nuclei degenerate. The cultures usually die after one sometimes after three weeks, even if the explantate is removed and the medium renewed.Under unfavourable conditions most cells do not emigrate; they often change colour. If cells emigrate they may have a threadlike starved or strongly vacuolized gorged appearance. Since the cells of the intermediate layer in situ do not differentiate into functionally diverse types, the cultivated cells probably start from the same physiological point. An inventory of the structural characteristics of in vitro cells is given. Certain characteristics are taken to serve as criteria for the quality of the culture fluids used.Results obtained from 664 explantates are reported. Culture fluids with high or low nutrient concentration, with or without hemolymph or egg extract are used (table p. 406–408).Salt solutions with monosaccharides and/or disaccharides are not sufficient. Protein hydrolysates (e.g. Vago) or amino acids — as present in larval hemolymph (Wyatt) — are required. A molar ration of Na K c. 0.2 1 is essential. The addition of hemolymph extract improves otherwise insufficient media. Egg extracts lead to hyperthrophy. The addition of vertebrate extracts and/or trehalose proved unsuccessful.The following factors are important for the culture of insect cells:A species and stage specific ratio of Na⁺ K⁺; the presence of sugar and protein derivatives; traces of certain substances which in our tests are supplied by extracts from the original organism.

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Mit bestem Dank für die Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Isolation and characterization of β-glucosidases from Aspergillus nidulans mutant USDB 1183

Two extracellular -glucosidases (cellobiase, EC 3.2.1.21), I and II, from Aspergillus nidulans USDB 1183 were purified to homogeneity with molecular weights of 240,000 and 78,000, respectively. Both hydrolysed laminaribiose, -gentiobiose, cellobiose, p-nitrophenyl--L-glucoside, phenyl--L-glucoside, o-nitrophenyl--L-glucoside, salicin and methyl--L-glucoside but not -linked disaccharides. Both were competitively inhibited by glucose and non-competitively (mixed) inhibited by glucono-1,5-lactone. -Glucosidase I was more susceptible to inhibition by Ag⁺ and less inhibited by Fe²⁺ and Fe³⁺ than -glucosidase II.     

Effects of photoperiod,temperature, food and relative humidity on the induction of diapause in the predatory miteAmblyseius potentillae

Aspects of the induction of diapause were studied in a Dutch strain of the phytoseiid miteAmblyseius potentillae. The photoperiodic response curve was of the long-day type, with a sharply defined critical daylength of 14.5 h. Critical daylength varied only little at temperatures between 15.0 and 22.5°C.All post-embryonic and possibly even late-embryonic stages of development were found to be sensitive to photoperiod; sensitivity appeared to be maximal during the protonymphal stage.It is shown that -carotene is necessary for some early step in the physiological mechanism of photoperiodic induction, and not (or not exclusively) for the expression of the diapause response.Two points of sensitivity to light could be demonstrated in the nights ofld 1311 andld 1212 long-night regimes, using 1-h night interruptions. These results are similar to those obtained in lightbreak experiments with spider mites and insects. However, no effect was found with light interruptions applied during the dark phase of anld 1014 long-night regime.In resonance experiments with a constant photophase (12 h) and a variable scotophase, a weak rhythmic response was found at 22.5°C; at 19.0°C this effect was completely absent.The relative humidity experienced by the mites during diapause induction as well as during diapause development influenced the rate of diapause completion under long days (ld 168). Diapause duration appeared to be shortest when the mites experienced low relative humidity (35±5%) during diapause induction and high relative humidity (75±5%) during diapause termination, and longest under the reverse conditions.     

Further evidence for abnormal protein kinase C regulation of macromolecule secretion in fibroblasts from cystic fibrosis patients

In comparison to skin fibroblasts from normal subjects, those from patients with cystic fibrosis (CF): (1) bound [20-³H] phorbol 12,13-dibutyrate (PDBu) with a higher affinity (K_d=25.8 vs 12.8 nM respectively) but expressed a similar number of total phorbol ester binding sites (about 2.5 pmol PDBu bound/mg of protein); (2) exhibited a faster and higher response to 4-phorbol 12-myristate 13-acetate (PMA) for the stimulation of [³⁵S]-labelled glycoconjutate release, but were equally sensitive to the synergistic effect of A23187 on this process; and (3) secreted glycoconjugates with similar [³⁵S]-sulfate and [¹⁴C]-leucine to [¹⁴C]-glucosamine labelling ratios. Taken together, these results provide further evidence for abnormal protein kinase C (PKC) regulation of macromolecule secretion in CF disease.Abbreviations BSA Bovine serum albumin - DBcAMP Dibutyryl cyclic AMP - DMEM Dulbecco's modified Eagle's medium - DMSO Dimethylsulfoxide - LDH Lactate dehydrogenase - PBS Phosphate-buffered saline - PDBu 4-phorbol 12,13-dibutyrate - 4-PDD 4-phorbol 12,13-didecanoate - PMA 4-phorbol 12-myristate 13-acetate - TCA Trichloroacetic acid     

Bewegungsstudien an Anatinen

Zusammenfassung Vorliegende Arbeit ist eine Weiterführung derLorenz'schen Bewegungsstudien an Anatinen aus dem Jahre 1941, fortgesetzt an Mischlingen zwischen den dort beschriebenen Arten. Die sich dabei ergebenden Befunde machten eine erneute Untersuchung der Elternarten notwendig. Außerdem wurden einige Arten beobachtet, deren Verhalten noch nicht untersucht worden war. Fragestellung und Begründung werden in der Einleitung gegeben.Im zweiten Abschnitt werden einige der vonLorenz gemachten Beobachtungen berichtigt. So zeigten einige der Kreuzungen mitbahamensis, daß die vonLorenz bei eben dieser Art als Kurzhoch-werden bezeichnete Bewegungsweise dem Ab-auf anderer Schwimmenten homolog ist. Ebenso ist die eine der beiden vonLorenz als Kurzhoch-werden bezeichneten Verhaltensweisen des Krickerpels als Ab-auf zu deuten. Der Gruß desflavirostre-Erpels wurde auch bei weiblichen Tieren gesehen. BeiAnas acuta wurde ein Kinnheben festgestellt, das sich in der Form stark vom Kinnheben beiplatyrhynchos unterscheidet. Als neue Verhaltensweisen wurden u. a. das Haltungannehmen und das Tendieren beimflavirostre-Erpel beschrieben.Im dritten Abschnitt werden einige Verhaltensweisen und ihre Funktion diskutiert und der Versuch gemacht, eine Motivationsanalyse zu geben.(Zeichnungen vonHermann Kacher)     

Ultrastructure of the tubule of the aglomerular teleost Nerophis ophidion

Summary The tubules in the aglomerular kidney of Nerophis ophidion are composed of cells showing different types of specializations of their plasma membrane. All cells possess a luminal brush border composed of microvilli, and show presence of vesicles with 100 Å thick unit membranes — some containing electron dense material —, tubular elements, multivesicular bodies, and plasma membrane invaginations in their apical cytoplasm. These features suggest an absorptive function of the cells. The apical portions of the cells are supplied with typical cilia.Some cells have abundant basilar plasma membrane invaginations usually lacking cytoplasmic organelles. Other cells appear to form interdigitating basilar cytoplasmic processes containing mitochondria; still other cells have smooth basilar cell membranes. These findings are discussed with reference to the known secretory function of the tubules and are compared with tubular fine structure in other species. It was concluded that urine formation by tubular secretion may occur in cells with different types of basilar cell membrane specializations.The occurrence of coated vesicles associated with invaginated basilar plasma membranes indicates transport of proteins (from peritubular blood vessels ?) at these sites.The tubule cells have abundant smooth surfaced endoplasmic reticulum and large and numerous active Golgi zones.Supported by grants from Fonden til Videnskabens Fremme and Therese och Johan Anderssons Minne. Part of this study was done at the Zoologica Stazione, Naples.The assistance of Miss Britt-Marie Petterson and Mr. Magnus Norman is gratefully acknowledged.