Determination of the threshold value of the mutagenic activity of Trenimon® on human lymphocytes “in vitro”

Summary The mutagenic activity of the tri-functional ethyleneimino derivate Trenimon^® (2,3,5-tri-ethyleneimino-benzoquinone 1,4) was tested in very low concentrations on human lymphocytes.The investigations revealed that a concentration of 2.8×10^-10 Mol/l or higher caused a marked dose-dependent increase of all types of aberrations. In concentrations less than 2.8×10^-10 Mol/l, the amount of aberrant cells as well as the rates of gaps and breaks per metaphase lay in the same range as for the blank control, as indicated by statistical treatment of the results.

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Zusammenfassung Bestimmung der Schwellendosis der cytogenetischen Wirkung von Trenimon^® an menschlichen Lymphocyten in vitro.Die cytogenetische Aktivität des trifunktionellen Äthyleniminderivates Trenimon^® (2,3,5-Tris-äthylenimino-benzochinon 1,4) in sehr niedrigen Konzentrationsbereichen wurde an menschlichen Lymphocytenkulturen geprüft.Die Versuche ergaben daß von der Konzentration 2,8×10^-10 Mol/l an ein steiler Anstieg aller Aberrationstypen mit der Dosiserhöhung zu verzeichnen ist. In den Konzentrations-bereichen unter 2,8×10^-10 Mol/l lagen die Anzahl aberranter Zellen sowie die Achromasie-und Bruchraten pro Metaphase im, Bereich der Nullkontrolle.Der experimentell ermittelte Schwellenwert der cytogenetischen Aktivität von Trenimon wurde durch die statistischen Teste erhärtet.

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This investigation comprises the results of a work pressented for a diploma by W. Kaufmann     

Die wirkung von Trypaflavin allein und in Kombination mit sichtbarem Licht auf die Chromosomen von HeLa-Zellen und menschliche Leukocyten

Summary Baldermann (1967) and Brucklacher (1968) investigated the effect of acridines on spermato- and oogenesis in mice. In both experimental series acridines were found mild mutagenes.This paper reports the induction of chromosomal aberrations in HeLa cells and cultured peripheral lymphocytes by the acridine trypaflavine (24 h of treatment). In addition the influence of visible light (1 h) was tested in these systems.In both cell types 10^-6 mol/l trypaflavine induces mainly chromatid gaps and breaks and reduces the mitotic rate. Light increases breakage of chromosomes in concentrations from 10^-7 to 10^-9 mol/l trypaflavine. Under these conditions there was also an increase of reunion figures in lymphocytes.Treatment with trypaflavine plus light lead to endomitosis and endoreduplication when chromosome preparations were made 24 h after the mutagen was removed from HeLa cells. In lymphocytes tetraploids appeared in experiments with 1 h of light 20 h before harvesting.It is supposed from the data that trypaflavine acts as a mutagenic agent to human cells, if the single cell is directly in contact with a concentration of 10^-6 mol/l.

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Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Über die Physiologischen Eigenschaften von Gewebekulturen aus Tumor- und normalem Gewebe vonPicea glauca

Summary In contrast to other plant tumor tissues (crown gall and genetic tumors), that fromPicea glauca can be cultivated in vitro only in the presence of IAA. The auxin requirement for maximal growth of the tumor is appreciably higher (10^–5 g/ml IAA) than that of normal Picea cultures (10^–8 g/ml). A similar marked difference is also found in the ascorbic acid requirement, which is 10^–5 g/ml for neoplastic, but 10^–7 g/ml for normal tissue. The significance of these results for the characterisation and the in vitro growth of plant tumors is discussed.

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Mit 4 Textabbildungen     

Rates of spontaneous mutation in certain genes of an arrhenotokous mite, Tetranychus pacificus

A number of 25.000 haploid males of Tetranychus pacificus was examined for the occurrence of mutations in certain genes for pigmentation. The spontaneous mutation rate in the six chosen genes p, al, we, w, le and st appeared to lie between 0.8×10^-4 and 2.8×10^-4. This high mutation rate is discussed in relation to the adaptational potentiality of the population, particularly with respect to insecticide resistance.

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Spontane mutationsraten bei gewissen gene der arrhenotoken milbe Tetranychus pacificus

Zusammenfassung 25000 haploide Männchen von Tetranychus pacificus wurden auf das Auftreten von Mutationen bei bestimmten Farbgenen geprüft. Die spontane Mutationsrate schien bei den Genen p, al, we, w, le und st zwischen 8×10^-4 und 2.8×10^-4 zu liegen. Diese hohe Mutationsrate wird im Hinblick auf die Anpassungsfähigkeit der Population besonders bezüglich der Insektizidresistenz diskutiert.

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The investigation is part of a study granted by the Shell Prize for Biology in the Netherlands.     

Zur Populationsgenetik der sauren Phosphatase der Erythrocyten (E C 3.1.3.2): Phänotypen-und Allelhäufigkeiten in Südwestdeutschland

Summary Within an population sample of 300 individuals of Southwestern Germany the red cell acid phosphatase polymorphism is investigated. Gene frequency estimates are: P^a=0.31, P^b=0.643, P^c=0.047.

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Direktor: Prof. Dr. med. Dr. H. Baitsch

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Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Die quantitative Erfassungsgrenze in der Histochemie

Zusammenfassung Durch Vergleich licht- und elektronenmikroskopischer Befunde am Nebennierenmark wird als praktische Erfassungsgrenze für Catecholamine 10^–11g Adrenalin pro Zelle ermittelt. Das entspricht einer Konzentration im Zytoplasma von 1,0%. Diese Konzentration wird auch als Verdünnungsgrenze in Modellversuchen festgestellt. Aufgrund theoretischer Überlegungen wird dargelegt, daß 10^–11 bis 10^–12g/Zelle allgemein die für eine beliebige histochemische Reaktion notwendige Konzentration darstellt.

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Summary Comparing light and electron-microscopical data from adrenal medulla, the identification limit for catecholamines has been calculated to be in the order of 10^–11g adrenaline/cell. This is equivalent to a concentration of 1,0% adrenaline/cell which has been found to be the dilution-limit in model experiments, too. On the basis of theoretical calculation it has been demonstrated that 10^–11–10^–12g/cell is the concentration needed for any histochemical reaction.

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Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Genetisch kontrollierte Varianten der NADH-Diaphorase

Zusammenfassung Die NADH-Diaphorase wurde an 725 gesunden Probanden mit Hilfe der Stärkegelelektrophorese untersucht. Zwei verschiedene Varianten wurden beobachtet: eine heterozygot schnelle (DIA 2-1) und eine heterozygot langsame (DIA 3-1). Die Genhäufigkeiten sind: DIA²=0,0021; DIA³=0,0007.

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Genetically determined variants of NADH-diaphorase

Summary By means of starchgel-electrophoresis a screening for variants of NADH-Diaphorase was carried out within a sample of 725 healthy probands. Two kinds of genetically determined variants have been observed: a heterozygous phenotype with greater mobility (DIA 2-1) and a heterozygous phenotype with slower mobility (DIA 3-1). The gene-frequencies are estimated so far as 0.0021 (DIA²) and 0.0007 (DIA³).

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Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Untersuchungen zur biologischen Aktivität der Phenylborsäure

Summary After 10 applications of phenylboric acid (0.03 ml of a 2·10^–3 M solution) to young plants ofKalanchoe blossfeldiana the flowers were found to have a reduced number of petals or no petals at all.InCucumis sativus one application of phenylboric acid at the same strength was enough to cause abnormal flowers to develop in the axils of 8 to 10 leaves; in addition there were 1 to 2 leafles nodes and other abnormalities. The position of the abnormal flowers and leafless nodes was dependant upon the stage of development of the plant at the time of application. It seems from a parallel study of treated plants grown to maturity, and plants sectioned at the time of treatment, that phenylboric acidinhibited selectively the first differentiation processes of the leaf primordia.

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Mit 9 Textabbildungen

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D 77.     

Mikroskopisch-anatomische Untersuchung der Veränderungen am Kopf des Hühnerkeimes nach TEM-Behandlung während der Organogenese

Ohne Zusammenfassung

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Malformations in the head of the chick after treatment with TEM during organogenesisI. Skeleton, musculature, brain, cranial nerves

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Herrn Prof. Dr. W. Kocher danke ich für Überlassung des Themas, Anregungen und Kritik.

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Teilweise unterstützt mit Mitteln aus den Euratomverträgen Nr. 041-65-10 BIO-D und Nr. 077-69-1 BIO-C.     

Zur Populationsgenetik der Adenylatkinase: Genhäufigkeit in einer südwestdeutschen Stichprobe

Summary Adenylate kinase phenotypes were determined in 407 unrelated persons from Southwestern germany. The frequencies were estimated to be AK¹=0.969, AK²=0.031.

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Direktor: Prof. Dr. med. Dr. H. Baitsch

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Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

In vitro-Verfahren zur Bestimmung der DNS-Synthese-Dauer einzelner Zellen Biochemische Voraussetzungen und Ergebnisse

Zusammenfassung Nach Hemmung der de novo-Synthese von TMP durch 5-F-UdR läuft die DNS-Synthese in vitro in Anwesenheit von 10^–5 M/l mit unveränderter Geschwindigkeit ab. Dadurch ist die DNS-Synthese-Dauer über die quantitative autoradiographische Messung des ¹⁴C-TdR-Einbaus bestimmbar. Diese in vitro-Methode wird an Rattenknochenmark und Zellen einer CML geprüft. Erste Ergebnisse zeigen eine gute Übereinstimmung mit in vivo-Messungen durch andere Methoden.

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In vitro determination of the duration of DNA synthesis of individual cells

Summary Suspended haemopoietic cells are incubated in the presence of 5-fluoro-deoxyuridine (5-F-UdR) as an inhibitor of the de novo synthesis of thymidine monophosphate (TMP). The DNA synthesis rate remains undisturbed, if 10^–5 M/l thymidine (TdR) is added. Thus, the DNA synthesis rate of individual cells can be determined by means of a quantitative autoradiographic method in measuring the incorporation rate of ¹⁴C-TdR into the DNA of the cells. DNA synthesis rates are then converted into DNA synthesis times. This in vitro method has been checked in cells of rat bone marrow and of peripheral blood in a case of chronic myelocytic leukaemia (CML). Preliminary results correlate well with in vivo estimations obtained by other methods.

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Studie im Rahmen des Assoziationsvertrages EURATOM-GSF für Hämatologie Nr. 031 641 BIAD

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Unterstützt durch die Deutsche Forschungsgemeinschaft: SFB 51/5     

Untersuchungen über da Auxin der Maiskoleoptile

Summary By means of theAvena section test in conjunction with paper chromatography very efficient growth inhibiting substances of acid nature, but no native auxins, could be demonstrated in ether extracts of Zea mays coleoptile tips. However it was possible to show that in the same extracts as much as 10^–5 g of added IAA was no longer demonstrable by either method. The significance of these results for the evaluation of rerent findings on the nature of auxins in Zea mays seedlings is discussed.

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Mit 2 Textabbildungen     

Die Phenoloxydasen des Ascomyceten Podospora anserina

Summary Laccase of the wild strain of Podospora anserina was purified by subsequent treatment with protamine sulfate, precipitation with ammonium sulfate and column chromatography on DEAE-Sephadex and hydroxylapatite. The purity was confirmed by sedimentation and electrophoresis (molecular weight about 361 000, isoeletric point at pH 5.1).The blue-coloured pure laccase has its absorption-maxima at 280 and 605 m. The substrate specifity of the enzyme corresponds to results which have been earlier obtained with unpurified preparations (Esser 1963 b). Laccase is very temperature sensitive. It loses its activity after both freezing and heat treatment (half-life time at 60°C about 6 min). The Michaelis-constants as determined with Dopa, potassium ferrocyanide and catechol are in the range of 2 to 5·10^-3 Mol/l. The appropriate value for ascorbic acid is about 10^-2 less. The laccase contains about 12% carbohydrate and about 7,5% nitrogen. According to its copper content of 0.123% the laccase carries seven atoms of copper per molecule.

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Teil I erschien in Arch. Mikrobiol. 46, 217–226 (1963).     

Mikrokalorimetrisch ermittelte Oszillationen beim Betriebsstoffwechsel von Hefen unter Sauerstoffdruck

Zusammenfassung In Glucosepuffer inkubierte Hefezellen zeigen unter Sauerstoffdruck (1 bis 10 kp/cm²) eine oszillierende Wärmeproduktion, die an die Atmungskompetenz der Zellen gebunden ist. Die Schwingungen werden auf periodische Schwankungen der mittleren Dichte der Zellen auf Grund veränderlicher Kohlendioxydkonzentration zurückgeführt.

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Microcalorimetric measurements of oscillations in the heat production of yeasts during glucose metabolism under oxygen pressure

Summary Yeast cells incubated in glucose buffer under oxygen pressure (1 to 10 kp/cm2) show an oscillating heat production which depends on the respiratory abilities of the cells. The oscillations are due to periodic density fluctuations of the cells by reason of variable carbon dioxide concentrations.

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Herrn Professor Dr. Dr. h. c. mult. B. Rajewsky zum 80. Geburtstag gewidmet.     

Zur Populationsgenetik der 6-Phosphogluconatdehydrogenasen (EC: 1.1.1.44): Genhäufigkeit in einer südwestdeutschen Stichprobe

Summary The 6-PGD phenotypes of 404 unrelated individuals from South-Western Germany were determined. The frequency of the 6-PG _B ^b1 allele is 0.031.

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Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Mechanismen der Aufnahme von Zink durch Bäckerhefe

Summary The uptake of labelled Zn by baker's yeast after exhaustion of the intracellular substrates for energy metabolism has been investigated. Without addition of glucose (substrate), equilibrium is reached rapidly. Binding of Zn is attributed to cell wall components. The amount bound approaches a saturation value at a Zn concentration in solution of the order of 10^-2 M. Initially, the Zn is bound reversibly, but it gradually changes into a firmly bound form. Substrate-independently, killed cells bind more Zn than living cells; apparently more binding sites become accessible. After addition of substrate, however, living cells take up additional Zn irreversibly over long periods. It is suggested that the Zn absorbed substrate-dependently enters the interior of the cell. The process does not depend strongly on the presence of air. It follows Michaelis-Menten kinetics, and the temperature coefficients (Q ₁₀), at different concentrations and temperatures, are 2–2,7. No efflux of Zn from the interior of the cell is observed. Reasons are given for considering the substrate-dependent uptake of Zn as an active process.

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Herrn Prof. Dr. O. Hromatka zum 65. Geburtstag in Verehrung gewidmet.     

Häufigkeit der Sichttypen in den Erbsystemen Haptoglobin,Gc, saure Erythrocytenphosphatase,Phosphoglucomutase und Adenylatkinase sowie den Erbeigenschaften Gm(1), Gm(2) und Inv(1) bei Deutschen (aus dem Raum Freiburg i. Br. und Köln) und bei Türken

Zusammenfassung Die an 274 Türken und einer mehrfachen Zahl Deutscher (787–5030) durchgeführten Untersuchungen ergaben eine etwas größere Häufigkeit des Gens Hp¹ im Raum Köln und eine größere des Gens Inv¹ im Raum Freiburg i. Br. Bei Türken sind die Allele Hp², Gm¹, P^b und PGM² häufiger, die Allele Hp¹, Gm^1,2, P_a, P_c und PGM¹ seltener als bei Deutschen. Die Frequenzen im Gc- und AK-System stimmen überein.

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Summary There was found a higher frequency of Hp¹ and a lower of Inv¹ in the population of Cologne than in the population of Freiburg. The frequencies of Hp², Gm¹, P^b and PGM² in the Turkish population were found to be higher than those in the German population; the frequencies of Hp¹, Gm^1,2, P^a, P^c and PGM¹ were found to be lower. In the systems Gc and AK the frequencies in the two populations are not significantly different.

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(Direktor: Prof. Dr. R. Haas)

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(Direktor: Prof. Dr. G. Pulverer)

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(Direktor: Prof. Dr. C. Bennholdt-Thomsen)     

Studies on the polymorphism of the red cell acid phosphatase in the population of Northern Bavaria (Germany)

Summary Within a population sample of 2000 blood donors of Northern Bavaria the AP polymorphism has been investigated. The calculated gene frequencies are: P^A=0.3463, P^B=0.5970, P^C=0.0568.

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Zusammenfassung An einer Bevölkerungsstichprobe von 2000 Blutspendern aus Nordbayern wurde der AP-Polymorphismus untersucht. Die errechneten Genfrequenzen sind: P^A=0,3463, P^B=0,5970, P^C=0,0568.

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With technical assistence of A. Melchertsen.     

Fibroblast culture in Werner's syndrome

Summary Growth capacities of fibroblasts from a woman with Werner's syndrome and of normal fibroblasts are compared by means of subculture-counting and ³H-thymidineuptake determination. The cells from the patient showed lower ³H-incorporation values.

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Zusammenfassung Bei Fibroblasten einer Patientin mit einem Werner-Syndrom und normalen Fibroblasten wurden die Reproduktionsrate und das Aufnahmevermögen für ³H-Thymidin in vitro verglichen. Beim Werner-Syndrom war die ³H-Aufnahme niedriger als bei den normalen Kontrollzellen.

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Supported by a grant from the Netherlands Organization for the Advancement of Pure Research, Fungo Branch.     

Stoffwechselprodukte von Mikroorganismen

Zusammenfassung Es wurde die Wirkungsweise von Borrelidin untersucht und mit zwei Makrolid-Antibiotica verglichen. Borrelidin hemmt in einem zellfreien System aus Bacillus subtilis spezifisch den Einbau von Threonin in sRNS bei einer Konzentration von 1,4·10^-6 m. Hingegen hemmen die Makrolid-Antibiotica Lankamycin (neutral) und Erythromycin (basisch) bei Konzentrationen, die zehnmal höher liegen als die wachstumshemmende Konzentration, nicht die Verknüpfung von Threonin oder anderer Aminosäuren mit sRNS.In Zellen hemmt Borrelidin bei einer Konzentration von 1,5–3·10^-6 m die Synthese von Protein, RNS und DNS. Lankamycin und Erythromycin hingegen hemmen nur die Synthese von Protein.Es wird vermutet, daß Borrelidin das erste Antibioticum ist, das die Protein-synthese über die Hemmung des Einbaus von Aminosäuren in sRNS blockiert.

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Metabolic products of microorganisms. 62nd information the inhibition of the attachment of threonine to sRNA in a cell-free system and of the sythesis of protein and nucleic acids in the cell by the antibiotic borrelidin

Summary The mode of action of borrelidin has been studied and compared with two macrolide antibiotics. Borrelidin specifically inhibits the attachment of threonine to sRNA at a concentration of 1,4·10^-6 m in a cell-free system of Bacillus subtilis. However, the macrolide antibiotics lankamycin (neutral) and erythromycin (basic), at concentrations which are ten times higher than those required to inhibit growth, still do not inhibit the binding of threonine or other amino acids to sRNA.In cells borrelidin inhibits at concentrations of 1.5–3·10^-6 m the synthesis of protein, RNA and DNA. In this point as well borrelidin acts differently from lankamycin and erythromycin which inhibit the synthesis of protein selectively.It is supposed that borrelidin is the first antibiotic inhibiting protein synthesis via amino acid attachment to sRNA.

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61. Mitt.: B. Maurer, A. Müller, W. Keller-Schierlein u. H. Zähner: Ferribactin, ein Siderochrom aus Pseudomonas fluorescens Migula. Arch. Mikrobiol. 60, 326–339 (1968).