A lectin from the liverwortMarchantia polymorpha L

Bryophytes have been screened for lectins. From the liverwortMarchantia polymorpha (Marchiantiales) a lectin could be purified to homogeneity using a combination of ultrafiltration, size exclusion chromatography and ion exchange chromatography. SDS polyacrylamide gel electrophoresis, size exclusion chromatography and electrospray mass spectroscopy showed that the lectin is a monomeric protein with a M_r of 16,134.64 ± 2.93.Marchantia polymorpha lectin agglutinates erythrocytes of different mammalia and exhibits carbohydrate specifity against complex carbohydrate structures. This is the first report of a lectin isolated from liverworts.This article forms Publication No. 71 of the Arbeitskreis Biologie und Chemie der Moose.     

Evidence that the prostaglandin-like substances fromPropionibacterium acnes are not identical with PGE2

Summary The prostaglandin-like substances (PLS) isolated fromP. acnes were investigated by reversed phase chromatography and gas chromatography-mass spectrometry. These analyses demonstrated that PLS were not identical with PGE₂, which supports a concept of PLS as a potential mediator of the inflammatory process in acne vulgaris.     

Influence of oxygen concentration on development ofDrosophila melanogaster

Summary 1st, 2nd, and early 3rd instarDrosophila larvae are extremely sensitive to 100% O₂ or 75% O₂/25% N₂ (at atmospheric pressure) whereas eggs, late 3rd instar larvae, and pupae are relatively insensitive under our exposure conditions. Eclosing flies exposed to an O₂ enriched environment consistently possessed 2 eye abnormalities: dark eye color and altered eye shape.Supported by NSF-URP grant SP1782684 and a Summer Faculty Fellowship to Harry Nickla.     

Intraluminal pressure and equivalent arterial wall tension

Résumé P=T/r, une loi de Laplace, est d'habitude appliquée pour exprimer la relation entre la pression intraluminale (P) et la tension intramurale (T) d'un vaisseau de rayonr. Sur un segment tubulaire, et sur une bande hélicale de l'aorte du lapin, nous avons mesuré deux valeurs équivalentes,P _50% etT _50%, obtenues lorsque la contraction causée par la noradrénaline est réduite de moitié. Ainsi la valeur der, calculée en utilisant l'equationP=T/r, est environ 75% de la valeur mesurée.     

Versuche zum enzymatischen Abbau der «gebundenen» gibberelline vonPharbitis purpurea

Summary Seeds and fruit walls ofPharbitis purpurea contain in addition to the free gibberellins A₈, A₃, A₅, more polar gibberellin-like substances, so-called bound gibberellins tentatively namedPharbitis and. After purification by thin layer chromatography, these 2 substances were treated with both Ficin and Emulsin in buffered solutions of pH 6.2. After treatment with Emulsin, the free gibberellins A₈ and A₅, after treatment with Ficin the gibberellins A₃ and A₅, were detectable in thin layer chromatograms.     

Occurrence and biosynthesis of 11 (R)-hydroxy-eicosatetraenoic acid (11-R-HETE) in the Caribbean soft coralPlexaurella dichotoma

Gorgonian soft corals from the Caribbean Sea are known to contain prostaglandin-like compounds as well as other products of arachidonic acid lipoxygenation, and the formation of the latter has been suggested to represent the first step in prostaglandin biosynthesis. Here we present evidence for the presence of 11-R-hydroxy-5Z, 8Z, 12E, 14Z-eicosatetraenoic acid (11-R-HETE), as well as of the enzyme responsible for its biosynthesis, in the Caribbean gorgonianPlexaurella dichotoma. Lipid extracts fromP. dichotoma were purified by conventional SiO₂ column chromatography followed by reversed phase high-pressure liquid chromatography (HPLC). These yielded a component having chromatographic and spectroscopic properties identical to synthetic 11-HETE. Electron impact mass spectrometric analysis of the acetoxy-, methyl ester derivative of the compound confirmed its identity as 11-HETE, while chiral phase HPLC of the methyl ester derivative showed that the stereochemistry of the alcoholic carbon atom wasR. Enzymatically active homogenates fromP. dichotoma were able to convert both unlabelled and [³H] arachidonic acid into 11-HETE. In vitro biosynthesis of the latter metabolite was also observed with homogenates of the Mediterranean gorgonianParamuricea clavata, another non-prostaglandin-containing soft coral, thus suggesting that 11-R-HETE production is not necessarily accompanied by prostaglandin formation in gorgonian corals.     

Purification and some properties of hemagglutinin from the Myxomycete,Physarum polycephalum

A new hemagglutinin was isolated from the plasmodium ofPhysarum polycephalum by salting out with ammonium sulphate followed by chromatography on DE-32, DEAE-Toyopearl and hydroxyapatite. This hemagglutinin, named physarumin, was purified 1000-fold over crude extracts. The molecular weight of physarumin was determined to be 22,000 by size exclusion chromatography on Bio-Gel P-60 and 8,700 by SDS-polyacrylamide gel electrophoresis. Physarumin agglutinated rabbit, guinea pig, horse and human erythrocytes. Physarumin-induced hemagglutination was inhibited by fetuin and ₁ glycoprotein, but not by commercially available mono-and disaccharides. Hemagglutinating activity was blocked by EDTA, and was restored by adding Ca²⁺ but not by Mg²⁺.     

Production of functional rat liver PSP protein in Escherichia coli

An efficient Escherichia coli expression system for the production of a perchloric acid-soluble protein (PSP) has been constructed. Complementary DNA encoding PSP was inserted into an inducible bacterial expression vector pGEX-4T-1. After the plasmid introduced into E. coli was expressed by isopropyl 1-thio-β-D-galactopyranoside (IPTG), the recombinant product was purified by glutathione-Sepharose 4B affinity chromatography. The purified product showed the expected NH₂-terminal sequence, but the translation inhibitory activity of this product was 10 times lower compared with that of authentic PSP isolated from rat liver. Received 8 October 1998; received after revision 6 November 1998; accepted 6 November 1998     

Photoreactivation of UV damage in culturedDrosophila cells

Summary Cell survival and photoreactivation of 254 nm ultraviolet (UV) light damage in a wild typeDrosophila cell line was assayed by colony formation in liquid medium. F_o, F_q, and extrapolation number for the exponential portion of survival curves are 21 J/m², 3.6 J/m², and 1.5 for non-photoreactivated cells and 110 J/m², 11.2 J/m², and 1.3 for those exposed to photoreactivating light. Maximal photoreactivation occurs at the 100 J/m² region of the curve. At 10 and 50% survival, 75–80% of the UV damage was photoreactivable.     

Resveratrol-type oligostilbenes from Iris clarkei antagonize 20-hydroxyecdysone action in the Drosophila melanogaster B(II) cell line

Bioassay-guided high-performance liquid chromatography analysis of a MeOH extract of Iris clarkei seeds yielded the resveratrol-type oligomeric stilbenes, ampelopsin B and α-viniferin, which antagonize the action of 20-hydroxyecdysone; with a 20-hydroxyecdysone concentration of 50 nM, the ED₅₀ values were 33 μM and 10 μM, respectively. The structures of these compounds were determined by spectroscopic analysis, notably ultraviolet, liquid secondary ion mass spectrometry and modern one- and two-dimensional nuclear magnetic resonance techniques. Received 4 November 1999; accepted 13 December 1999     

Chemical defense in the three European species ofCrematogaster ants

The composition of the Dufour gland of the antC. scutellaris has been reinvestigated by gas chromatography/mass spectrometry. The major components of the gland are (2E,5E,12Z)-4-oxoheneicosa-2,5,12-trien-1-ol acetate (1a) its ¹⁴ and ¹⁶ double bond isomers (1b and1c), and the corresponding (Z,Z)-dienes5a and5b, all containing an acetylated C₂₁ chain. The previously proposed structures1d, 1e, and5c, which are based on an homologous acetylated C₂₃ chain, correspond to minor derivatives present in the gland. Traces of acetylated C₁₉ homologs, tentatively identified as1g-1i, have also been found. The Dufour gland contents of the two other EuropeanCrematogaster species have also been studied.C. auberti is very similar toC. scutellaris in producing mainly1a, 1b and1c, together with the same higher and lower homologs, but it lacks the dienic derivatives5, whereasC. sordidula contains essentially the acetylated C₁₉ compounds1g, 1h, and1i, accompanied by acetylated C₁₇ homologs.     

Biological characterization of a new and highly potent synthetic analogue of corticotrophin

Zusammenfassung Ein ACTH-Analogon:d-Ser¹-Nle⁴-(Val-NH₂)²⁵--1-25-Corticotrophin wurde tierexperimentell sowie orientierend humanpharmakologisch untersucht. Das Pentacosapeptid (DW-75) zeigt auf Grund der Substitution von 3 Aminosäuren eine gegenüber Peptiden mit natürlicher Sequenz beachtlich erhöhte ACTH-Aktivität.     

Partial purification and characterization of acid phosphatase from sporulated oocysts ofEimeria tenella

Summary Acid phosphatase ofEimeria tenella oocysts (Peak II) was purified 77-fold with a recovery of 26% using protamine sulfate precipitation, DEAE-cellulose chromatography and Sephadex G-200 gel filtration. This enzyme occurs in multiple forms as indicated by two peaks which can be separated by DEAE-cellulose chromatography and polyacrylamide gel electrophoresis. The partially purified enzyme has optimal activity at pH 4.5. With p-nitrophenyl phosphate the Km and V_max values for (Peak II) were 25 mM and 1.57 mol/min/mg protein, respectively. The enzyme (Peak II) ist strongly inhibited by Hg⁺⁺, Cu⁺⁺, iodoacetamide, fluoride and molybdate. Tartrate and other divalent metal ions have no effect on enzyme activity. The partially purified Peak II phosphatase is not a glycoprotein as it is not absorbed on concanavalin-A Sepharose and its treatment with bacterial neuraminidase does not alter its elution profile through DEAE cellulose.     

Chemical analysis of the pheromone blends produced by males and females of the neotropical moth,Mocis megas (Guénée) (Lepidoptera,Noctuidae, Catocalinae)

Summary Pheromonal secretions produced by females and males of the noctuid moth,Mocis megas (Guénée) have been analyzed by gas chromatography and mass spectrometry (EI (electron impact) and CI (chemical ionization)). The female sex pheromone was a blend of (Z,Z,Z) 3,6,9 heneicosatriene (55%) and (Z,Z) 3,6-cis-9S, 10R-epoxyheneicosadiene (45%). Male secretion produced at the level of a prothoracic organ was a blend of two unsaturated major hydrocarbons: (Z,Z) 6,9 heneicosadiene, (64%) and (Z,Z,Z) 3,6,9 heneicosatriene (24%) and C₁₉, C₂₀ and C₂₂ homologues (total ratio 12%), as minor components. The trienic hydrocarbon was present in both sexes. The behavioral role of this male secretion has not yet been elucidated.     

Metabolism of amphibian spermatozoa in relation to their motility

Summary Bufo arenarum spermatozoa were able to sustain motility both under aerobic and anaerobic conditions. In aerobiosis, the oxygen consumption varies between 2.6 and 4.2 l O₂/10⁸ cells/h at 30°C. The synthesis of lactic acid by anaerobic spermatozoa demonstrated the existence of an active glycolytic pathway.This work was supported by the Consejo Nacional de Investigaciones Científicas y Técnicas and the Latinoamerican Program of Human Reproduction (PLAMIRH), grant 34-95-2-75.     

Affinity chromatography of human serum proteins using matrix bound lectin fromViscum album L.

Summary The D-galactose specific lectin fromViscum album L. reacts with serum proteins that contain the corresponding D-galactopyranosyl residues. By affinity chromatography of human serum on lectin-sepharose IgM, ₂-macroglobulin, haptoglobin and -lipoprotein were quantitatively retained. Only parts of IgA, IgG and transferrin were retarded. The other serum proteins are unbounded as albumin, ₁A– and ₁C.     

The dimeric and co-operative myoglobin ofNassa mutabilis. A peculiar case

The myoglobin present in the radular muscle of the Prosobranchia sea snailNassa mutabilis is a peculiar case among myoglobins. It is a dimer showing co-operative oxygen binding equilibrium curves with pO_{2 1/2}=4.7 mmHg, invariant with pH, and n=1.6. Although the globin is composed of 147 amino acid residues, corresponding to a molecular mass of 15760 D, gel filtration chromatography of the native myoglobin indicate Mr=26000±2000 D. Similarly, acrylamide electrophoretic analyses in SDS and velocity sedimentation indicate a molecular mass of about 13000 D for the denatured globin. The molecule is highly unstable and forms slowly a chromogen when aged or immediately upon oxidation to the ferric state. The visible region of the absorption spectrum of the O₂ or CO liganded myoglobin derivatives indicate an altered heme environment. Circular dichroism analyses confirm this indication showing negative Cotton effects in all regions of the heme absorption spectra of the MbO₂ and MbCO derivatives. Interestingly, the CD spectrum of the oxidised met-form shows a positive band almost symmetrical with respect to that of the MbO₂ derivative. This is similar to what reported for the monomeric hemoglobin ofGlycera dibranchiata for which a reversed heme orientation was proposed. Detailed resonance Raman spectroscopic studies have permitted a more direct investigation of the interactions between the heme and the protein. The proximal Fe-Im bond shows a stretching mode frequency down shifted by 5 cm^–1 with respect to the corresponding band of horse heart myoglobin, in good correlation with the much higher instability ofNassa m. myoglobin and its much lower oxygen affinity. The unusual bond instability finds additional support in a kinetic study in which the myoblogin is mixed with CO in buffered solutions at different pH values. This approach gives evidence that the Fe-Im bond is broken upon lowering the pH, with a pK of 4.0±0.2, the highest among those of deoxy hemoproteins. The rupture of the proximal bond appears to occur with a proton-linked transition showing n=1.8±0.1, again indicating cooperativity between the two subunits. The vinyl and propionate heme substituents show resonance Raman spectroscopic bands indicating different modes of interaction with their environment with respect to other myoglobins. Most interestingly, the vinyl stretching mode frequency, typically a single band, appears split in two bands inNassa m. myoglobin. This splitting is evident in all the investigated derivatives of the myoglobin, indicating that vinyl 2 and 4 are not equivalent in this molecule. A similar splitting has been found so far only inChironomus t.t. hemoglobin.     

Negative adsorption of anions by suspensions of bentonites

Résumé Nous avons étudié l'adsorption négative des anions I*O ₃ ^– et Cr*O ₄ ^–– à différents bentonites en fonction d'électrolytes additionnés. Pour les électrolytes NaCl et Na₂SO₄ les résultats sont en bon accord avec ceux deSchofield etMattson; nous avons étudié aussi l'effet de l'addition de solutions de CaCl₂; celles-ci produisent le même effet à des dilutions 20 fois plus grandes que les solutions de NaCl. Comme ceci ne peut pas être expliqué sur la base de la théorie deDebye-Hückel, nous avons appliqué la théorie exacte de la couche double; le facteur 20 mentionné plus haut mène à une valeur de=–75 mV. En introduisant cette valeur de, une représentation graphiques–1/c pour les résultats des expériences au Na-bentonite + solutions de NaCl, donne une surface spécifique de 1110 m²/g.     

Die Dünnschichtchromatographie von Gibberellinen

Summary Thin-layer chromatography on silica gel is described as a new method for the separation and identification of the gibberellins A₁, A₃, A₄, A₅, A₇, A₈, and A₉.