Isolation and characterization of neurokinin A, neurokinin A(3-10) and neurokinin A(4-10) from a neutral water extract of a metastatic ileal carcinoid tumour

A metastasis to the right liver lobe of an argyrophil/argentaffin midgut carcinoid tumour in a patient with the classical carcinoid syndrome was examined for the presence of tachykinins other than substance P, using a specific antiserum. The extract was initially purified using SepPak cartridges, and subsequently subjected to cation-exchange chromatography on SP Sephadex C-25 which separated the immunoreactive material into two main components (components I and II). Both were further purified by anion-exchange chromatography on DEAE-Sephadex A-25, and by reverse-phase fast protein liquid chromatography. Component II was identified as neurokinin A by its immunochemical and chromatographic properties and amino acid sequence analysis. Component I consisted of two molecular forms which were identified as neurokinin A(3-10) and neurokinin A(4-10) by amino acid sequence analysis. The tumour tissue contained only small amounts of the eledoisin-like peptide that has earlier been demonstrated in mammalian tissues. Although this component behaved like the nonmammalian peptide eledoisin on reverse-phase HPLC and on reverse-phase ion-pair chromatography, eledoisin-specific antiserum E2 indicated that eledoisin-like peptide is not identical to eledoisin. Neurokinin A in carcinoid tumours has an N-terminal heterogeneity; this multiplicity constitutes a further support for the hypothesis that carcinoid tumours produce a number of tachykinins which may be present in different relative amounts in individual patients and may contribute to the individual differences in symptomatology.     

Surface properties of lactobacilli isolated from the small intestine of pigs

One hundred wild-type strains of the genus Lactobacillus were isolated from the small intestine of newly-slaughtered pigs up to 6 months of age. Cell surface hydrophobicity and capsule formation were studied on a number of strains. Strains showing high surface hydrophobicity as measured by the salt-aggregation test and hydrophobic interaction chromatography on Octyl Sepharose were commonly found to adhere in high numbers to isolated pig intestinal epithelial cells. Heat and protease treatment of bacteria of high surface hydrophobicity, including autoaggregating strains in phosphate-buffered saline, showed a drastic decline in this surface property. Three hydrophilic strains (LBp 1044, 1068 and 1073) also showed binding to intestinal cells but at a lower level (approx. 5 bacteria/cell) as compared with the best binding hydrophobic strain (LBp 1063, approx. 11 bacteria/cell). These findings suggest that different or multiple adhesion mechanisms may be involved in the colonization of the small intestinal mucosa of pigs. Cultures of selected strains grown in liquid media rich in carbohydrates did not affect their hydrophobic cell surface character. Therefore it seems less likely that carbohydrate capsule polymers are the major determinants of intestinal colonization of lactobacilli in pigs.     

Enzyme action in polymer and salt solutions. I. Stability of penicillin acylase in poly(ethylene glycol) and potassium phosphate solutions in relation to water activity

The stability of penicillin acylase (penicillin aminohydrolase, EC 3.5.1.11) was studied in poly(ethylene glycol) and potassium phosphate solutions. Enzyme stability measured as the half-life of the enzymatic activity and the transition temperature determined by differential scanning calorimetry, correlated well. The enzyme stability could not be related to the water activity as a measure of solute-solvent interaction. It seems to be related more to the concentration of the solutes and much less to the molecular weight of poly(ethylene glycol). The stabilizing effect of poly(ethylene glycol) is also discussed in terms of poly(ethylene glycol)-protein interactions.     

A morphometric and ultrastructural study of lithium-induced changes in the medullary collecting ducts of the rat kidney

Summary Rats were given a lithium-containing diet (40 mmol/kg) to Study the effect of lithium on the structure of collecting ducts from the inner stripe of the outer medulla. The results show that there is a significant increase in the volume density of collecting ducts already after one week on this diet. The volume density of both intercalated and principal cells increases, whereas the volume density of mitochondria in the cytoplasm increases in the intercalated cells only. The increased volume of both principal and intercalated cells seems to be part of a general hyperplasia and hyperactivity of the collecting duct, which may in some way be related to the effects of lithium on vasopressinmediated water transport. The specific changes in the intercalated cells may be a consequence of the effects of lithium on distal nephron potassium and hydrogen ion transport in the distal nephron.     

Further antigenic analyses of the fish pathogenic bacteriumVibrio anguillarum

TenVibrio anguillarum strains were selected for an immunoelectrophoretic study. Evidence was provided for existence of two new K antigens which displayed cross-reactivity. The importance of an exact characterization of surface antigens inV. anguillarum is considered.     

Aqueous two-phase partition coefficients for Escherichia coli β-galactosidase fusions

The partition of native Escherichia coli -galactosidase and of two different fusion proteins comprised mainly of -galactosidase from E. coli was studied in aqueous two-phase systems composed of polyethylene glycol (PEG) and dextran. These fusions contain an amino-terminal segment from the E. coli outer membrane protein F (OmpF) and a linker peptide. Differences in the partition pattern could be observed for the three enzymes despite their similarity. Decreased polymer concentrations in the phase system increased the partition coefficient for all three -galactosidases.     

Host plant recognition in monophagous weevils: specificity in feeding responses of Ceutorhynchus constrictus and the variable effect of sinigrin

Host plant relations of the monophagous weevil Ceutorhynchus constrictus Marsh. (Coleoptera: Curculionidae: Ceutorhynchinae) feeding on garlic mustard, Alliaria petiolata (Bieb.) Cavara & Grande (Cruciferae) were studied in the laboratory. Most other crucifers were rejected in choice tests using garlic mustard as a reference plant, but Brassica nigra, Sinapis alba and Thlaspi arvense were as acceptable as the host plant. Flowering plants of Descurainia sophia were acceptable while young plants of this species were not. The most important feeding stimulants in extracts of garlic mustard were uncharged, water soluble compounds. The most abundant glucosinolate in garlic mustard, sinigrin, was a feeding stimulant, too. However, the feeding stimulatory activity of sinigrin was only expressed in the presence of still unidentified uncharged compounds from garlic mustard leaves. Host plant relations in monophagous crucifer-feeding insects is discussed in relation to the distinctness of glucosinolate patterns found in their host plants.

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Zusammenfassung Ceutorhynchus constrictus Marsh. (Coleoptera: Chrysomelidae: Ceutorhynchinae) ist ein monophager Rüsselkäfer, der an Knoblauchhederich frisst. Das Wirtswahl-Verhalten dieses Käfers ist im Labor untersucht worden. Die meisten Crucifiren waren im Wahlversuche nicht akzeptiert, wenn Knoblauchhederich als Vergleichspflanze vorhanden war. Von Brassica nigra, Sinapis alba, und Thlaspi arvense wurden im Vergleich gleiche Mengen verzehrt wie von der Wirtspflanze. Blühende Descurainia sophia Pflanzen wurden, im Gegensatz zu Jungpflanzen der gleichen Art, angenommen. Die wichtichsten Phagostimulanten in Extrakten von Knoblauchhederich-Blättern waren ungeladene, wasserlösliche Substanzen. Das häufigste Glukosinolat im Knoblauchhederich, Sinigrin, war auch ein Phagostimulant. Doch war die phagostimulierende Wirkung von Sinigrin nur in Kombinationen mit noch nicht identifizierten, ungeladenen Substanzen aus Knoblauchhederich-Blätter nachweisbar. Wirtspfanzen-Beziehungen von monophagen Insekten werden diskutiert im Zusammenhang mit der Eigenart des Glukosinolat-Inhaltes ihrer Wirtspflanzen.

     

The effect of glucosinolates on responses of young Phyllotreta nemorum larvae to non-host plants

All recorded host plants of Phyllotreta nemorum L. (Coleoptera: Chrysomelidae) contain glucosinolates and belong to the plant families Brassicaceae (Cruciferae), Resedaceae and Capparaceae. The acceptability of 56 plant species from 28 other plant families (non-hosts) for young larvae has been studied in the laboratory. None of these species were fully acceptable for initiations of leaf mines when intact untreated leaves were presented, and only one species, Malva silvestris L. (Malvaceae), was partially acceptable. The acceptability of some species increased when leaf discs were presented instead of intact leaves; but the highest percentages of mine initiations occurred in leaf discs treated with the glucosinolate, sinigrin. A stimulatory effect of sinigrin could be demonstrated in experiments with 7 plant species: Papaver dubium L., Papaver rhoeas L., Fumaria officinalis L., Malva silvestris L., Pisum sativum L., Campanula latifolia L. and Lactuca sativa L. The majority of species remained unacceptable even after treatment with glucosinolates.The main causes for these differences between plant species are supposed to be differences in contents of deterrents and/or other stimulants for mine initiation. These possibilities are discussed in relation to the content of allelochemicals in acceptable plants and the position of these plants in botanical classifications.

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Zusammenfassung Phyllotreta nemorum L. ist ein oligophager Erdfloh, der an Cruciferen und anderen Glukosinolat-haltigen Pflanzenarten gebunden ist. Die Imagines fressen Löcher in die Blätter und die Weibchen legen ihre Eier in den Boden. Die Larven sind Blattminierer. Nach dem Schlüpfen im Boden klettern sie an die Pflanzen hoch, und die Einbohrung und der Anfang der Minierung erfolgt in eines der unteren Blätter der Wirtspflanze.Die Wirkung von Glukosinolaten auf die Einbohrung von Junglarven in Pflanzenarten, die keine natürliche Inhalt von Glukosinolaten haben, ist in Laborexperimenten untersucht worden. 56 Pflanzenarten aus 28 Familien wurden präsentiert teils als unbehandelte Blätter und teils als Glukosinolatbehandelte Blattscheiben. Unbehandelte Blätter von nicht-Glukosinolathaltigen Arten waren immer unbefriedigend für die Larven. Nur in eine Art, Malva silvestris L. war die Frequenz der Einbohrung ein bisschen höher als 10%. Eine signifikante Erhöherung der Anzahl eingebohrten Larven nach der Sinigrin-behandlung erfolgte in 7 Pflanzenarten: Papaver dubium L., P. rhoeas L., Fumaria officinalis L., Malva silvestris L., Pisum sativum L., Campanula latifolia L. und Lactuca sativa L. Doch blieben die meisten Pflanzenarten (84%) auch nach der Sinigrin-Behandlung unbesiedelt.Pflanzenarten, die nach der Sinigrin-Behandlung nicht besiedelt werden enthalten vielleicht frasshemmende Stoffe, oder ihnen fehlen noch weitere Frass-stimulierende Stoffe. Diese Möglichkeiten werden diskutiert in Zusammenhang mit den Inhalt von Allelochemikalien in besiedelten Pflanzenarten und mit ihrer taxonomischer Position.

     

Bacterial Metabolism of 2,6-Xylenol

Strain DM1, a Mycobacterium sp. that utilizes 2,6-xylenol, 2,3,6-trimethylphenol, and o-cresol as sources of carbon and energy, was isolated. Intact cells of Mycobacterium strain DM1 grown with 2,6-xylenol cooxidized 2,4,6-trimethylphenol to 2,4,6-trimethylresorcinol. 4-Chloro-3,5-dimethylphenol prevents 2,6-xylenol from being totally degraded; it was quantitatively converted to 2,6-dimethylhydroquinone by resting cells. 2,6-Dimethylhydroquinone, citraconate, and an unidentified metabolite were detected as products of 2,6-xylenol oxidation in cells that were partially inactivated by EDTA. Under oxygen limitation, 2,6-dimethylhy-droquinone, citraconate, and an unidentified metabolite were released during 2,6-xylenol turnover by resting cells. Cell extracts of 2,6-xylenol-grown cells contained a 2,6-dimethylhydroquinone-converting enzyme. When supplemented with NADH, cell extracts catalyzed the reduction of 2,6-dimethyl-3-hydroxyquinone to 2,6-dimethyl-3-hydroxyhydroquinone. Since a citraconase was also demonstrated in cell extracts, a new metabolic pathway with 2,6-dimethyl-3-hydroxyhydroquinone as the ring fission substrate is proposed.     

Fractionation of immunoglobulins by liquid-liquid partition chromatography in aqueous two-phase systems

In this paper we show that although immunoglobulins are easily precipitated in solutions containing polyethylene glycol (PEG), especially at pH's where the conformation of the proteins should be close to native, human and rabbit IgG can be solubilized in aqueous dextran/PEG two-phase systems containing glycine and sodium chloride at pH 7.0 and that human IgA and IgM can be solubilized in such systems if the pH is increased to 9.0. Liquid-liquid partition chromatography (LLPC) on Li-ParGel was used to separate immunoglobulins into subfractions. Human IgG, IgM, and IgA all gave three peaks in the system used. These results indicate the possibility of separating different classes of immunoglobulins with this method. Specific IgG antibodies isolated from a rabbit antiserum against human serum proteins gave only two peaks in the LLPC system while the total IgG population gave three, as did human IgG. Thus, partitioning of immunoglobulins seems to be related to antibody activity.