C4 polymorphism in the dog: Molecular heterogeneity of the C4α and C4γ subunit chains

Using an immunoblotting technique and goat antihuman C4, we observed five distinct electrophoretic variants of C4 in a panel of 60 random dogs. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of immunoprecipitated C4 showed that dog C4 is composed of three polypeptide subunit chains (, , and ) and that structural variability occurs within the - and -chain regions. Two distinct molecular weight forms of both the C4- ( _A and _B) and C4-( _A and _B) chain were detected. The variant forms of C4 and C4 were found in association with particular C4 allotypes.     

Darstellung und Charakterisierung eines Lipoproteins mit Antigenwirksamkeit im Lp-System

Zusammenfassung Unter Anwendung verfeinerter Trennverfahren wurde das -Lp(a+)-Lipoprotein des menschlichen Serums in immunologisch und elektrophoretisch reiner Form dargestellt. Es wurde hinsichtlich seiner Lipidzusammensetzung analysiert und mit ebenfalls immunologisch und elektrophoretisch rein erhaltenen -Lp(a-)- und ₁-Lipoproteinen verglichen.

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Separation and characterization of a lipoprotein with antigenic activity in the Lp-system

Summary Through application of refined separation techniques the -Lp(a+)-lipoprotein of human serum was characterized in an immunologically and electrophoretically pure form. It was analyzed with regard to its lipid-composition and compared to likewise electrophoretically and immunologically pure -Lp(a-)- and ₁-lipoproteins.

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Untersuchung mit Unterstützung durch die Deutsche Forschungsgemeinschaft     

Stimulation of phospholipase A2 by auxin in microsomes from suspension-cultured soybean cells is receptor-mediated and influenced by nucleotides

The molecular mechanism of membrane-associated reactions induced by auxin was investigated in membranes isolated from cultured cells of soybean (Glycine max L.). Auxins increased the activity of phospholipase A₂ in microsomes isolated from suspensioncultured soybean cells. The reaction was measured as the accumulation of radioactive lysophosphatidylcholine hydrolyzed from radioactive phosphatidylcholine in membranes which had been prelabelled with [¹⁴-C]choline in vivo. Stimulation by auxin was detectable after 1 min and was auxin-specific in that weak auxins had little effect. Auxin concentrations as low as 2·10^–8 M and up to 2·10⁺³ M -naphthaleneacetic acid already stimulated the phospholipase A₂ activity. Guanosine and adenosine diphosphate at 100 M, if applied during homogenization of cells, completely abolished the stimulation of phospholipase A₂ by auxin and, when applied after homogenization, had no effect. Guanosine and adenosine 5-O-thiotriphosphate, uridine 5-diphosphate, and uridine 5-triphosphate, all at 100 M, had no effect in either treatment, suggesting that only nucleotides entrapped in the vesicles could exert an effect. The effect of auxin on phospholipase A₂ had an optimum at pH 5.5 and was abolished completely by an antibody against the membrane-associated auxin-binding protein from maize coleoptiles, applied after homogenization. This antibody recognized a 22-kDa polypeptide in highly purified plasma membranes from cultured soybean cells. This suggests a receptor function for this auxin-binding protein and a role for a cytosolic nucleotide-binding protein in the activation of phospholipase A₂ by auxin. It is concluded that phospholipase A₂ has a function in plant signal transduction.Abbreviations ABP auxin-binding protein - ATP S adenosine 5-O-thiotriphosphate - 2,4-D 2,4-dichlorophenoxyacetic acid - GTP S guanosine 5-O-(thiotriphosphate) - IgG immunoglobulin G - LPC lysophosphatidylcholine; - -NAA , -naphthaleneacetic acid - PLA₂ phospholipase A₂ We cordially acknowledge the gift of anti-ABP antibody by D. Klämbt and the help by H. Ordowski (both Botanisches Institut, Universität Bonn) with the immunoblotting experiments. This work was supported by the Deutsche Forschungsgemeinschaft.     

Isolation and characterization of a Forssman antigen-binding lectin from velvet bean (Mucuna derringiana) seeds

A Forssman antigen (GalNAc1-3GalNAc1-3Gal1-4Gal1-4Glc1-1Cer)-binding lectin has been purified from velvet bean (Mucuna derringiana) seeds by a combination of affinity chromatography and reversed phase HPLC. This lectin agglutinates both native and trypsin-treated sheep erythrocytes as well as trypsinized rabbit erythrocytes, but neither native rabbit nor human erythrocytes, irrespective of blood group type. SDS-PAGE and gel filtration chromatography reveal the lectin to be a homodimer consisting of two 54 kDa subunits linked by non-covalent bonds. The results obtained by quantitative precipitation, haemagglutination inhibition and TLC overlay assays indicate that theMucuna lectin specifically recognizes Forssman antigen and Forssman disaccharide (GalNAc1-3GalNAc)-related structures. Abbreviations: The abbreviations and the trivial names used are: AH, 6-aminohexyl; BSA, bovine serum albumin; Cer, ceramide; HPLC, high performance liquid chromatography; PAGE, polyacrylamide gel electrophoresis; PBS, 10mm phosphate-buffered saline, pH 7,2, containing 0.15m NaCl; PMSF, phenyl methyl sulfonyl fluoride; SDS, sodium dodecyl sulphate; TFA, trifluoroacetic acid; TBS, 20mm tris-buffered saline, pH 7.2; TLC, thin-layer chromatography; A disaccharide, GalNAc1-3Gal; A trisaccharide, GalNAc1-3[Fuc1-2]Gal; Forssman disaccharide, GalNAc1-3GalNAc; CDH (ceramide dihexoside or lactosyl ceramide) Gal1-4Glc1-1Cer (LacCer); CTH (ceramide trihexoside or globotriosyl ceramide), Gal1-4Gal1-4Glc1-1Cer (GbOse₃Cer or Gb₃); globoside (globotetraosyl ceramide), GalNAc1-3Gal1-4Gal1-4Glc1-1Cer (GbOse₄Cer or Gb₄); Forssman antigen (globopentaosyl ceramide), GalNAc1-3GalNAc1-3Gal1-4Gal1-4Glc1-1Cer (GbOse₅Cer).     

Zur Überwindung der biologischen Grenze Meer—Land durch Mollusken

Zusammenfassung Die Überwindung der biologischen Grenze zwischen Meer und Land durch Mollusken ist bisher nur wenig im Hinblick auf physiologische Umstellungen untersucht worden. Unter diesem Aspekt wurden Experimente durchgeführt an Alderia modesta, einem amphibisch lebenden Opisthobranchier des unteren Supralitorals, und an Ovatella myosotis, einer Pulmonate, die im oberen Supralitoral vorkommt. Alderia zeigt eine relativ enge Salinitätstoleranz. Zu hohe und zu niedrige Salzgehalte hemmen die Entwicklung, führen zu Mißbildungen bei der Embryonalentwicklung und lassen adulte tiere rasch eingehen. In Beziehung zu der Salinität der angrenzenden Gewässer (Brackwasser der Ostsee und der Ästuare, Meerwasser der Nordsee) existieren verschiedene Biotypen, die auf Aussüßung und hohe Salzkonzentration unterschiedlich reagieren.Die weiter ins Supralitoral vorgedrungene Ovatella myositis hat eine wesentlich breitere Salinitätstoleranz. Schädigungen der Adulten treten auf Süßwasser nicht, auf Salzwasser erst oberhalb von 55 auf. Durch schrittweise Adaptation können höhere Konzentrationen ertragen werden. Der Toleranzunterschied zwischen den untersuchten Populationen ist genetisch bedingt. Eiablage, Embryonalentwicklung und Wachstum erfolgen im Bereich zwischen 5 und mindestens 50 Substratsalinität.Physiologisch wird diese große Salinitätstoleranz durch ein fast durchgehendes poikilosmotisches Verhalten ermöglicht. Auf sehr ausgesüßtem Substrat sind die Schnecken stark hypertonisch (auf Süßwasser hat das Binnenmedium auf 7), im ganzen übrigen Bereich weniger stark (3–4 über der Substratsalinität). Im oberen Extrembereich vermögen die Tiere diese Hypertonie offenbar nur schwer aufrechtzuerhalten (vgl. S. 149).Die Veränderungen von Gastropoden auf dem Wege vom Meer über das Supralitoral zum Land werden diskutiert; im Gebiet der Salzwiesen läßt sich ein günstigeres Schema zu dieser Frage aufstellen als bei den bisher meist zitierten Littorinen.

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On the emigration of gastropodes from the sea: Studies on Alderia modesta and Ovatella myosotis

Summary Up to now little attention has been paid to the phylogenetic emigration of gastropods from the sea: almost nothing is known about the physiological changes which enable snails to live in terrestrial habitats. To help solve this question, experiments were carried out on Alderia modesta, an opisthobranch slug of the lower supralittoral, and Ovatella myositis, a primitive pulmonate snail of the upper supralittoral. In each case two subspecies were studied.North Sea Alderias differ from their Baltic counterparts in size and salinitytolerance. The difference in size between Ovatella of the Mediterranean and of the Baltic is very slight. There is a marked difference, however, in the salinity-tolerance. Alderia modesta survives in a comparatively narrow range of different salinities. The optimum is between 10 and 20 for Baltic specimens and between 15 and 35 for those of the North Sea. The results are the same in and out of water. Ovatella can live on a freshwater substratum as well as on 55. Specimens of the Mediterranean can even become adapted to 90. Flooded with water of differrent salinities the tolerated range becomes markedly smaller. Eggs are produced between 5 and 50 (Baltic specimens) or 5 and 65 (Mediterranean specimens). All eggs develop without being damaged between 5 and 40 and 10 and 45 respectively. The optimal salinity-concentration for growth and egg-production is 10. Salinity-concentrations below and above these marks disturb the development. In one spawn-mass some eggs cleave normally, others become deformed, and again others do not cleave at all. This heterogeneous reaction points to the existence of something like a physiological polymorphism in regard to salinity-tolerance. The salinity-concentration of the blood of Ovatella was measured after a long-term acclimatisation. Ovatella is poikilosmotic and slightly hypertonic (3–4) throughout almost the whole range of salinities it tolerates. On extremely low concentrations it becomes more hypertonie; on extremely high concentrations it becomes nearly isotonic.

     

The chromosomal localization of human β-galactosidase revisited: a locus for β-galactosidase on human chromosome 3 and for its protective protein on human chromosome 22

Summary A series of man-Chinese hamster and man-mouse somatic cell hybrids was investigated to study the localization of the genes coding for the human lysosomal enzyme -galactosidase (EC 3.2.1.23) and for its protective protein. Using a monoclonal antibody, raised against human placental -galactosidase, it was observed that the structural locus for the -galactosidase polypeptide is located on chromosome 3. The nature of the involvement of chromosome 22 in the expression of human -galactosidase was elucidated by metabolic labelling of the hybrids with radioactive amino acids, immunoprecipitation with monoclonal and polyclonal antibodies against -galactosidase, followed by analysis via gel electrophoresis and fluorography.The data show that the presence of chromosome 22 coincides with the presence of a 32 kd protein. This polypeptide, the protective protein was previously shown to be intimately associated with human -galactosidase. In addition, the protective protein was found to be essential for the in vivo stability of -galactosidase by aggregating -galactosidase monomers into high molecular weight multimes. Both chromosome 3 and 22 are therefore necessary to obtain normal levels og -galactosidase activity in human cells.     

The macromolecular composition of Geotrichum candidum grown on whiskey distillery spent wash

Summary Biomass harvested from batch and single-stage continuous cultures of the fungus Geotrichum candidum, growing on the spent wash from an Irish malt whiskey distillery, was analysed for macromolecular components. The true protein content was on average more than 47% (w/w), while crude protein levels of as much as 61% were found. The protein was not deficient in any amino acid when compared to the FAO standard. The effects of varying growth temperature and dilution rate on the composition of the organism are discussed, and also the problem of its accumulation of copper from the medium.     

Spatiotemporal expression patterns of sialoglycoconjugates during nephron morphogenesis and their regional and cell type-specific distribution in adult rat kidney

The expression of 2,6- and 2,3-linked sialic acids on N-glycans was studied in embryonic, postnatal, and adult rat kidney. Histochemistry and blotting using Polyporus squamosus and Sambucus nigra lectins for 2,6-linked sialic acids and the Maackia amurensis lectin for 2,3-linked sialic acids were performed and sialyltransferase activity was assayed. N-glycans with 2,6- and 2,3-linked sialic acid were differently expressed in the two embryonic anlagen and early stages of nephron. Metanephrogenic mesenchyme was positive for 2,3-linked sialic acid but not for the 2,6-linked one, which became detectable initially in the proximal part of S-shaped bodies. Collecting ducts were positive for 2,6-linked sialic acid, whereas 2,3-linked sialic acid was restricted to their ampullae. Although positive in embryonic kidney, S1 and S2 of proximal tubules became unreactive for 2,3-linked sialic acid in postnatal and adult kidneys. In adult kidney, intercalated but not principal cells of collecting ducts were reactive for 2,3-linked sialic acid. In contrast, 2,6-linked sialic acids were detected in all cells of adult kidney nephron. Blot analysis revealed a different but steady pattern of bands reactive for 2,6- and 2,3-linked sialic acid in embryonic, postnatal, and adult kidney. Activity of 2,6 and 2,3 sialyltransferases was highest in embryonic kidney and decreased over postnatal to adult kidney with the activity of 2,6 sialyltransferase always being three to fourfold that of 2,3 sialyltransferase. Thus, 2,6- and 2,3-linked sialic acids are differently expressed in embryonic anlagen and mesenchyme-derived early stages of nephron and show regional and cell type-specific differences in adult kidney.     

C-banding patterns in the AustralianBulbine (Liliaceae): The annual group,B. semibarbata s. lato

Chromosome C-band patterns have been studied in 34 populations of the Australian annualBulbine group, which comprises 4x (2n = 26, 28), 8x (2n = 52, 54) and 12x (2n = 78) populations. The 2n = 26B. semibarbata populations have a simple, low heterochromatin pattern with very minor polytypic variation. The 2n = 28 populations, corresponding morphologically to a group given separate status asB. alata, are similar in pattern but exhibit pronounced enhancement of telomeric and, more particularly, centromeric dot bands. NOR heterochromatin and satellites are difficult to identify inB. alata but appear to occur in different positions from the 26-chromosome karyotype. Eastern Australian 8 x patterns are consistent with a proposed hybrid ancestry,B. semibarbata ×B. alata. Annual and perennial C-band profiles in the AustralianBulbine are discussed briefly in relation to the additive and transformation models of heterochromatin evolution and to the possible adaptive significance of variation in heterochromatin content.Cytoevolution in the AustralianBulbine 2; for part 1 see Pl. Syst. Evol.157, 201–217.     

The T→C mutation at position +96 of the untranslated region 3′ to the terminating codon of the β-globin gene is a rare polymorphism that does not cause a β-thalassemia as previously ascribed

We have observed a TC mutation at position +96 of the untranslated region 3 to the terminating codon of the -globin gene in members of two Czech families and one black family. Data from initial studies suggested that this change was the cause of a -thalassemia, but continued analyses have provided convincing evidence that this mutation is a simple polymorphism.     

Enzymatic transformation of desacetyl-lanatoside A into digitoxin by β-glucosidase action in cyclodextrin solutions

Summary The enzymatic transformation of desacetyl-lanatoside A (DLA) to its secondary glycoside, digitoxin, in solutions of -and -cyclodextrins is effected using of -glucosidase from barley. Due to the interaction of cyclodextrins (CyDs) with desacetyl-lanatoside A, an increase in solubility of the latter of 24.5 and 230 times was observed for -cyclodextrin and -cyclodextrin, respectively. Kinetic studies of the enzymatic transformation gave for -glucosidase the values K_M=3.3×10^–4 mol. dm^–3 and V_max=0.557 mol mg^–1 min^–1 when the substrate was the deacetyl-lanatoside A complex with -cyclodextrin, while in the case of the complex with -cyclodextrin these values were K_M=5.45×10^–4 mol dm^–3 and V_max=0.896 mol mg^–1 min^–1.     

A conceptual model of ecosystem restoration triage based on experiences from three remote oceanic islands

A conceptual model, that illustrates restoration, ecological landscaping, rehabilitation and regreening, is developed. It considers biocentric, historical, aesthetic and engineering aspects. The term ecosystem restoration triage is used because the first step is to decide whether to do nothing (because, on the one hand, the system is too degraded to warrant restoration, or, on the other, because biological integrity is relatively intact and therefore either none, or minimal, restoration is required) or to do something (because restoration is worthwhile, urgent and feasible). This approach hinges on the definition that restoration in the strictist sense is a biocentric activity that returns the original compositional, structural and functional diversity, along with its dynamics and natural evolutionary potential. Original is a difficult qualifier as it depends on just how far back in time we go. Where human values are involved, this is not restoration in the pure sense of restoring ecological integrity, but is ecological landscaping, rehabilitation or regreening. Experience from three remote oceanic islands [Easter Island, Cousine Island (Seychelles), Marion Island (Sub-Antarctic)] and which represent near extremes of this model are used to illustrate it.     

A mathematical model of the interactions in the mixed culture of invertebrates and algae in the “producer–consumer” aquatic biotic cycle

This paper presents a mathematical model of interactions between two herbivorous invertebrates (ciliate Paramecium caudatum and rotifer Brachionus plicatilis) and two planktonic algae (Chlorella vulgaris and Scenedesmus quadricauda) spatially segregated in two compartments of a chemostat–type experimental microcosm system. The model mimics a producer–consumer aquatic biotic cycle, describing the dynamics of the mixed culture of ciliates and rotifers, as consumer compartment, feeding on the mixed algal culture, as producer compartment, under N-limiting conditions. We experimentally found that metabolites of the alga Scenedesmus produce an adverse effect on the reproduction of ciliate Paramecium. Taking this effect into account improved the behavior of the model, the results of which came into qualitative agreement with the experimental results. Both our experimental and modeling approaches demonstrated that, even in conditions of a spatially–segregated producer-consumer biotic cycle, species coexistence is impossible either in the mixed algal culture or in the mixed invertebrate culture. Scenedesmus excluded Chlorella, whereas Brachionus excluded Paramecium.     

Remyelination In Vitro Following Protein Kinase C Activator-Induced Demyelination

In previous work we found that mezerein, a C kinase activator, as well as basic fibroblast growth factor (FGF-2) induce demyelination and partial oligodendrocyte dedifferentiation in highly differentiated aggregating brain cell cultures. Here we show that following protein kinase C activator-induced demyelination, effective remyelination occurs. We found that mezerein or FGF-2 caused a transient increase in DNA synthesis following a pronounced decrease of the myelin markers myelin basic protein and 2,3-cyclic nucleotide 3-phosphohydrolase. Both oligodendrocytes and astrocytes were involved in this mitogenic response. Within 17 days after demyelination, myelin was restored to the level of the untreated controls. Transient mitotic activity was indispensable for remyelination. The present results suggest that myelinating oligodendrocytes retain the capacity to reenter the cell cycle, and that this plasticity is important for the regeneration of the oligodendrocyte lineage and remyelination. Although it cannot be excluded that a quiescent population of oligodendrocyte precursor cells was present in the aggregates and able to proliferate, differentiate and remyelinate, we could not find evidence supporting this view.     

Effect of mutation on DNA-composition of some bacteria

The aim of this study was to investigate whether DNA from bacteria would be drastically changed by spontaneous and induced mutation. Melting points of pure DNA were taken as a criterion to indicate possible changes. DNA from dark-blue Chromobacteria and from some pale-blue and non-pigmented spontaneous variants had the same melting point, indicating only a very small overall change in its base composition. A similar situation held forAcetobacter aceti (liquefaciens) and an Mn-induced mutant. A wild-typeAgrobacterium tumefaciens B6 and its streptomycin-resistant mutant S1 behaved similarly. However, pure DNA from another, UV-induced, mutant M39 had a melting point which was 1.5 C higher, showing that considerable alterations in the structure of DNA may occur upon mutation. The results are discussed.     

Light adaptation in cone photoreceptors: The occurrence and significance of unitary adaptive strength

The behavior of peak response-log intensity functions generated by the dark glasses model is examined and is shown to describe previously observed light adapted behavior in cone photoreceptors. The models of Boynton and Whitten (1970) and Norman and Werblin (1974) are closely related to the dark glasses model — the Boynton-Whitten model being more specific and the Normann-Werblin model more general. For the models, a certain parameter relationship will produce systems which have optimal intensity discriminative capacities. When the data are fitted, this parameter relationship — unitary adaptive strengh — seems to emerge. Possible evolutionary and psychophysical implications are discussed.     

The effect of dibutyryl cyclic AMP on the expression of actin isoforms in astroglia

Mammalian cells contain at least 8 actin isoforms. The functional significance and the mechanisms that regulate the expression of each actin isoform are not yet known. Using immunofluorescence staining, it was found that all astroglia in tissue culture express -actin isoform and about 86% of astroglia express -smooth muscle actin isoform. When astroglia were treated with dibutyryl cyclic AMP for 4,7,14 and 21 days, it was found that the number of the cells expressing -smooth muscle actin isoform progressively decreased, whereas, the number of the cells expressing -actin isoform remained constant. The western blot experiment showed that the amount of total -smooth muscle actin isoform (soluble and insoluble) and of the insoluble isoform expressed by astroglia treated with dibutyryl cAMP decreased whereas, the amount of total and insoluble -actin isoform expressed by the same cells did not show any significant changes. The cells treated with the cAMP failed to migrate and to close the area created by the scratch wound in monolayer culture. However, the non-treated cells migrated and closed the area created by the scratch after 3 days. This study shows that the astroglia have different mechanisms in regulating the expression of different actin isoforms and that the -sm actin isoform is important in migration of astroglia.     

Ganglioside-cholera toxin interactions: a binding and lipid monolayer study

Summary On t.l.c. plates ¹²⁵I-cholera toxin binds to a disialoganglioside tentatively identified as GD_lb with about 10 times less capacity than to ganglioside GM₁. Binding of labeled toxin to both gangliosides was abolished in presence of excess amounts of unlabeled B subunit. Ganglioside extracts from human or pig intestinal mucosa showed toxin binding to gangliosides GM₁ and GD_1b. In ganglioside-containing lipid monolayers the penetration of the toxin was independent of the ganglioside binding capacity.Abbreviations GM₂ Gal-NAc14Gal(3-2NeuAc)14G1c1Cer - GM₁ Gal3Ga1-NAc14Gal(32NeuAc)14G1c11Cer - GD_1a NeuAc23Ga113Gal-NAc14Gal(32NeuAc)14G1c11Cer - GD1b Gall3Gal-NAcl4Gal(32NeuAc82NeuAc)14Glc11Cer - GT_1b NeuAc23Ga113Ga1-NAcal4Gal(3-2NeuAc82NeuAc)14G1c11Cer - dpPC 1,2-hexadecanoyl-sn-glycero-3-phosphocholine - dpPE 1,2-hexadecanoyl-sn-glycero-3-phosphoethanolamine     

Reaction on alternating GC oligonucleotide templates

Summary The condensation reactions of activated nucleotides, ImpN or 2-MeImpN, with the selfcomplementary ribo-octanucleotide GCGCGCGC or with the partially self-complementary heptanucleotide GCGCGCG were studied. The templatedirected reaction of 2-meImpC with the heptamer yields the 3–5 octamer as the main product. All other reactions yield 2–5-linked octamers and pyrophosphates as major products. Surprisingly, 2-MeImpG facilitates the reaction of 2-MeImpC with the heptamer.Procedures for the analysis by gel electrophoresis of the oligomeric products obtained in reactions of this kind are described.     

An alternative propagation method ofAnanas through nodule culture

A micropropagation scheme forAnanas comosus Merr. was developed using nodule culture. Nodules were induced from leaf-base or chopped shoot-base explants on modified half-strength MS medium supplemented with 2.69-5.37 M NAA and 4.44 M BA and could be maintained long-term as nodules. The nodules proliferated into more nodules when chopped into pieces of 1–3 mm and placed onto the same medium. They regenerated shoots when transferred to medium supplemented with 0.54–10.74 M NAA and 0.44–8.88 M BA. The regeneration capacity of nodules is higher than that of direct regeneration or callus. Maximum regeneration was obtained from culture medium containing 0.54 M NAA and 0.44 M BA, where shoots could be observed as early as within 2 weeks. Many shoots formed roots in the same medium in which they were regenerated after 10 subcultures, but the best rooting occurred in medium containing 0.54 M NAA and 0.44 M BA. Rooted plantlets ofA. comosus Merr. could be routinely produced at 6-week intervals.Abbreviations NAA naphthaleneacetic acid - BA 6-benzylaminopurine