Apoptotic cell-derived factors induce arginase II expression in murine macrophages by activating ERK5/CREB

Apoptotic cell (AC)-derived factors alter the physiology of macrophages (MΦs) towards a regulatory phenotype, characterized by reduced nitric oxide (NO) production. Impaired NO formation in response to AC-conditioned medium (CM) was facilitated by arginase II (ARG II) expression, which competes with inducible NO synthase for l-arginine. Here we explored signaling pathways allowing CM to upregulate ARG II in RAW264.7 MΦs. Sphingosine-1-phosphate (S1P) was required and acted synergistically with a so far unidentified factor to elicit high ARG II expression. S1P activated S1P₂, since S1P₂ knockdown prevented ARG II upregulation. Furthermore, ERK5 knockdown attenuated CM-mediated ARG II protein induction. CREB was implicated as shown by EMSA analysis and decoy-oligonucleotides scavenging CREB in RAW264.7 MΦs, which blocked ARG II expression. We conclude that AC-derived S1P binds to S1P₂ and acts synergistically with other factors to activate ERK5 and concomitantly CREB. This signaling cascade shapes an anti-inflammatory MΦ phenotype by ARG II induction.     

Avilamycin

Summary A strain ofStreptomyces viridochromogenes produced a new crystalline antibiotic, Avilamycin, related to but not identical with Curamycin and Exfoliatin. Avilamycin, C₆₃H₉₄O₃₅Cl₂, gave on solvolytic degradation the following products: dichloroisoeverninic acid, 2-deoxy-d-rhamnose, 2,6-di-O-methylmannose, 4-O-methylfucose,l-lyxose and 3,5-diacetoxy--caprolactone.     

Über Struktur und Aktivität der den H2O2-Zerfall katalysierenden Cu2+-Komplexe. — VI. Differenzierung von nativer RNS und nativer DNS bzw. denaturierter DNS auf Grund der katalytischen Eigenschaften

Summary The catalysis of H₂O₂ decomposition by Cu²⁺-complexes of RNA and DNA has been investigated. It is shown that both complexes decompose H₂O₂, but only the Cu²⁺-RNA-system shows peroxidative activity too, e.g. only in this case the nucleotide bases are degraded. Thermal denaturation of DNA also leads to a Cu²⁺-complex with peroxidative activity, the latter being dependent on the degree of denaturation.

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V. Mitteilung:S. Petri, H. Sigel undH. Erlenmeyer, Helv. chim. Acta49, 1778 (1966).

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12. Mitteilung überMetallionen und H ₂ O ₂; 11. Mitteilung:H. Ch. Curtius, P. Anders, R. Zell, H. Sigel undH. Erlenmeyer, Helv. chim. Acta49, 2256 (1966).     

Inhibition of tyrosine degradation in vivo by the dopa decarboxylase blocking agent NSD-1034

Zusammenfassung Der Stoffwechsel von H ₃ ² -3, 5-l-Tyrosin wurde in mit NSD-1034 (100 mg/kg) vorbehandelten Versuchstieren (Hähnchen, katzen) untersucht. Als Folge des durch NSD-1034 stark gehemmten Abbaus entstanden während der Versuchszeit (ca. 20 min) nahezu kein Tritiumwasser und keine aromatischen Abbauprodukte. Auch die sonst zu beobachtenden radioaktiven Metabolite des H³-Tyrosins (Glutamin- und Asparaginsäure, Dopamin und Noradrenalin) waren nicht nachweisbar. Ausserdem war der Gehalt an H³-Tyrosin in Organen behandelter Tiere höher als in Kontrollen.     

Role of nitric oxide in the functional response to ischemia-reperfusion of heart mitochondria from hyperthyroid rats

We investigated the role of nitric oxide (NO) in the mitochondrial derangement associated with the functional response to ischemia-reperfusion of hyperthyroid rat hearts. Mitochondria were isolated at 3000 g from hearts subjected to ischemia-reperfusion, with or without N-nitro-L-arginine (L-NNA, an NO synthase inhibitor). During reperfusion, hyperthyroid hearts displayed tachycardia and low functional recovery. Their mitochondria exhibited O₂ consumption similar to euthyroid controls, while H₂O₂ production, hydroperoxide, protein-bound carbonyl and nitrotyrosine levels, and susceptibility to swelling were higher. L-NNA blocked the reperfusion tachycardic response and increased inotropic recovery in hyperthyroid hearts. L-NNA decreased mitochondrial H₂O₂ production and oxidative damage, and increased respiration and tolerance to swelling. Such effects were higher in hyperthyroid preparations. These results confirm the role of mitochondria in ischemia-reperfusion damage, and strongly suggest that NO overproduction is involved in the high mitochondrial dysfunction and the low recovery of hyperthyroid hearts from ischemia-reperfusion. L-NNA also decreased protein content and cytochrome oxidase activity of a mitochondrial fraction isolated at 8000 g. This and previous results suggest that the above fraction contains, together with light mitochondria, damaged mitochondria coming from the heaviest fraction, which has the highest cytochrome oxidase activity and capacity to produce H₂O₂. Therefore, we propose that the high mitochondrial susceptibility to swelling, favoring mitochondrial population purification from H₂O₂-overproducing mitochondria, limits hyperthyroid heart oxidative stress.Received 24 March 2004; received after revision 9 June 2004; accepted 5 July 2004     

Bactericidal activity againstPseudomonas aeruginosa is acquired by cultured human monocyte-derived macrophages after uptake of myeloperoxidase

Myeloperoxidase (MPO) is an enzyme located within polymorphonuclear neutrophils capable of producting cytotoxic oxidant species that are particularly active against bacteria with polysaccharide capsules.Pseudomonas aeruginosa (10⁶ bacteria per 1 ml) are killed within 1 h in vitro by a MPO/H₂O₂/Cl^– system (48 mU=132 ng of MPO). The question arose as to whether human macrophages would acquire cytotoxic activity when loaded with this enzyme. Monocytes were therefore isolated from human blood and cultured for up to ten days to induce maturation to macrophages. These cells lost endogenous MPO within five days while H₂O₂ production in response to stimulation by phorbol myristate acetate (10^–6M) decreased to 23% within ten days. On the other hand, their capacity to take up exogenous MPO increased fourfold from day three to day ten. Human macrophages cultured from eight days (when both H₂O₂ production and MPO uptake were sufficient) were therefore used to study the effects of MPO uptake on cytocidal activity againstPseudomonas aeruginosa. After a 1 h MPO loading period, macrophages (5×10⁵ cells per ml) were incubated in the presence of bacteria (0.5 to 2×10⁶ bacteria per ml) for 2 h at 37°C. At a bacteria/macrophage ratio of 1, only 34.8±7.0% of bacteria survived (compared to killing by non-loaded macrophages), while 74.4±9.3% survived at a ratio of 4. From these results, we conclude that loading macrophages with exogenous MPO could enhance their microbicidal activity, suggesting a potentially useful therapeutic application.     

Isomériecis-trans des ménaquinones et oxydations phosphorylantes dans les extraits deMycobacterium phlei

Summary Incubation ofM. phlei washed cells with [¹⁴C³H₃]-l-methionine led to [2-¹⁴C³H₃] dihydromena-quinone-9 with an isotope ratio identical to that of methionine. Chromatography of the doubly labelled quinone indicated, despite a pronounced isotope effect, that bothcis andtrans isomers had the same isotope ratio. This result eliminates any possibility of hydrogen exchange in the 2-methyl group of menaquinones during oxydative phosphorylation, even in thecis isomer. Furthermore, it is confirmed that this compound is certainly formed from natural or synthetic menaquinones during isolation or incubation periods by the effect of daylight irradiation.     

Incorporation of [14C] amino acids into liver S-RNA of chick. Effect of orotic acid and vitamin B12

Riassunto La somministrazione dietetica di vitamina B₁₂ e di acido orotico determina un aumento della incorporazione dellal-serina C¹⁴ nello RNA solubile del fegato di pulcino carente di B₁₂, mentre non è in grado di influenzare quella dellal-leucina¹⁴C e dellal-metionina¹⁴C.     

The structure of a new phytoecdysone kaladasterone: An application of13C magnetic resonance spectroscopy to structural problems

Zusammenfassung Isolierung und Strukturaufklärung von Kaladasteron (C₂₇H₄₂O₇), eines neuen Phytoecdysons, werden beschrieben.

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Acknowledgement: We would like to express our gratitude to Dr.P. Beynon and Dr.S. Murakami (Jeol Co., London and Milan) for running the CMR-spectra on the PS 100 PFT Spectrometer. We are also greatly indebted to Dr.S. Maroni for letting us have the high resolution mass spectrum on the Varian Mat 311 instrument.     

H<Subscript>2</Subscript>S biosynthesis and catabolism: new insights from molecular studies

Hydrogen sulfide (H₂S) has profound biological effects within living organisms and is now increasingly being considered alongside other gaseous signalling molecules, such as nitric oxide (NO) and carbon monoxide (CO). Conventional use of pharmacological and molecular approaches has spawned a rapidly growing research field that has identified H₂S as playing a functional role in cell-signalling and post-translational modifications. Recently, a number of laboratories have reported the use of siRNA methodologies and genetic mouse models to mimic the loss of function of genes involved in the biosynthesis and degradation of H₂S within tissues. Studies utilising these systems are revealing new insights into the biology of H₂S within the cardiovascular system, inflammatory disease, and in cell signalling. In light of this work, the current review will describe recent advances in H₂S research made possible by the use of molecular approaches and genetic mouse models with perturbed capacities to generate or detoxify physiological levels of H₂S gas within tissues.     

Comparison of some biological activities of arginine vasotocin and synthetic analogs

Zusammenfassung Nachweis, dass die Substitution in Positionen 3 und 8 im Arginin Vasotocin ([8-Arginin]-Oxytocin) selektive Veränderungen verschiedener biologischer Aktivitäten hervorruft, was am Modell der Oxytocinkonformation diskutiert wird.

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Supported in part by USPHS grant No. AM-13567.

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Neurohypophyseal hormone analogs are denoted in accordance with IUPAC-IUB Tentative Rules on Biochemical Nomenclature, Biochemistry6, 362 (1967) and J. biol. Chem.247, 977 (1972). All optically active amino acids are of thel-configuration unless otherwise noted. Arginine vasopressin, [8-arginine]vasopressin; lysine vasopressin, [8-lysine]vasopressin; AVT, arginine vasotocin, [8-arginine]oxytocin; Mpr, -mercaptopropionic acid.

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Acknowledgment. We thank Dr.Klaus Lübke, Schering AG, West Germany, for generously supplying us with the hormone analogs.     

Interference with histamine and imidazole acetic acid metabolism by salicylates: A possible contribution to salicylate analgesic activity?

Summary In man, rats and mice, the urinary excretion of the histamine andl-histidine metabolite, imidazole acetic acid, is increased and that of the conjugated metabolite, ribosylimidazole acetic acid, decreased by small doses of salicylates. In contrast to salicylates, other non-salicylate anti-inflammatory drugs, indomethacin, phenylbutazone, phenacetin and acetaminophen do not influence the excretion of the urinary metabolites of histamine andl-histidine. Since imidazole acetic acid is reported to have analgesic and narcotic activity, there is the inference that the analgesic properties of salicylate might be due in part to interference in imidazole acetic acid metabolism.     

Nitric oxide mediates histamine induced down-regulation of H<Subscript>2</Subscript> receptor mRNA and internalization of the receptor protein (R1)

During agonist-dependent long-term stimulation of cells, histamine receptor subtypes are frequently down-regulated. However, the mechanisms underlying the modulation of receptor expression during long-term histamine stimulation have yet to be resolved. Based on our recently reported results showing an H₁-mediated down-regulation of histamine H₂ receptor mRNA in endothelial cells, our aim was to characterize the mechanism controlling rapid and long-term histamine-mediated modulation of H₂ receptor expression in more detail. We were able to show that the histamine-induced down-regulation of H₂ receptor mRNA and cell surface expression lasting for 24 h was accompanied by augmentation of the receptor protein level in the cytoplasmatic fraction of endothelial cells for this time period. Furthermore, changes in receptor protein levels in whole-cell lysate were negligible, indicating that the rapid and prolonged modulation of cell surface H₂ receptor levels by histamine was regulated solely via internalization. The role of nitric oxide (NO) as a key mediator in histamine-stimulated cell responses was underlined by subsequent studies showing the attenuation of histamine-induced H₂ receptor mRNA down-regulation and protein trafficking following NO synthase isozyme inhibition.Received 11 March 2003; received after revision 11 June 2003; accepted 17 June 2003     

Sur le mycoside Cm,nouveau peptido-glycolipide isolé deMycobacterium marianum

Summary Mycoside C_m, m.p. 198–200° []_D = – 31° (CHCl₃), a new peptido-glycolipid isolated fromMycobacterium marianum, has the approximate molecular formula C₁₀₈H₁₈₅N₇O₂₈ and gives on hydrolysis seven molecules ofd-amino acids (oned-phenylalanine, threed-allo-threonine, threed-alanine), three molecules of 6-deoxyhexoses (one 6-deoxy-talose of as yet undetermined configuration and two molecules of 3,4-di-O-methyl-l-rhamnose); the lipid moiety of mycoside C_m is a hydroxy-acid of approximate formula C₅₀H₉₆O₃; three O-acetyl groups are also present. The preliminary formula (I) is proposed for mycoside C_m.

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60ème communication sur les constitutants des Mycobactéries; 59ème comm. voir: ref. 7.

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Ce travail a bénéficié d'une subvention de la Fondation Waksman pour le développement des Études microbiologiques en France, et d'une subvention du National Institute of Allergy and Infectious Diseases (Grant E 28 38).     

An anti-Bcl-2 antibody prevents 2-deoxy-D-ribose-induced apoptosis in the IPLB-LdFB insect cell line

Confocal microscopy reveals that the anti-Bcl-2 antibody (pAb) is able to diffuse across the plasma membrane of the fat body cell line IPLB-LdFB from the insect Lymantria dispar, demonstrating the presence of Bcl-2-like molecules in the cytoplasm. Immunoperoxidase procedure confirms the cellular localization. Furthermore, an immunoprecipitation corresponding to a molecular weight of 29 KDa is observed with western blot analysis using the anti-Bcl-2 pAb. Cytofluorimetric experiments show that anti-Bcl-2 pAb counteracts 2-deoxy-d-ribose-induced apoptosis and provokes morphological changes in the insect cell line, i.e. a reduction in cell size, the disappearance of the vacuola and changes in shape. At the same time, the antibody provokes mitochondrial membrane depolarization, and N-acetyl-l-cysteine is unable to reconstitute the physiological conditions. The present findings suggest that Bcl-2-like proteins play a main role in maintaining of the integrity of cellular components, e.g. mitochondria, rather than in controlling programmed cell death. Received 23 January 2001; received after revision 1 March 2001; accepted 1 March 2001     

The chemistry of the polymyxin antibiotics

Zusammenfassung Die Chemie der Polymyxine, einer Klasse von basischen Polypeptid-Antibiotika, begann 1954, als durch Gegenstromverteilung erstmals ein definierter Vertreter, das Polymyxin B₁ in reiner Form isoliert werden konnte (L. C.Craig). Partialhydrolyse mit Mineralsäuren führte zum Schluss, dass es sich um Cyclohepta- oder Cyclooctapeptide mit Seitenketten handelt, die, -Diaminobuttersäure (Dab) enthalten (W.Hausmann).Amidartige Verknüpfung der Seitenkette mit einer Fettsäure [(+)-6-Methyloctansäure (MOA) oder 6-Methylheptansäure (IOA)] (S.Wilkinson) verleiht diesen Antibiotika den Charakter von Invertseifen. Sie sind besonders gegen gramnegative Erreger wirksam. Schwierigkeiten bereiteten die Aufklärung der Verknüpfungsweise der Seitenkette (- oder-) und die Beantwortung der Frage, ob das Molekül nebend-Phenylalanin in der Ringsequenz noch einend-, -Diaminobuttersäurerest in Nachbarschaft zur Fettsäure in der Seitenkette enthält. Synthetische Versuche mitd-, -Diaminobuttersäure an dieser Stelle führten zu hochaktiven Produkten, die aber mit natürlichem Polymyxin B₁ nicht identisch waren. Entscheidende Fortschritte wurden mit dem bakteriellen Enzym Nagarse (T.Suzuki) erzielt, das schrittweise die Seitenkette bis zum Ringpeptid abbaut. Dabei ergab sich, dass den Polymyxinen die allgemeine Struktur eines Cycloheptapeptides mit-verknüpfter Seitenkette zukommt (Figur 4).Die Polymyxine B₁ E₁ (Colistin A) sowie Circulin A unterscheiden sich voneinander nur durch eine Variation in der gleichen Dipeptidsequenz des 7-gliedrigen Ringes. Die im Polymyxin B₁ vorhandene Dipeptidsequenzd-Phe-l-Leu ist in Polymyxin E₁ (Colistin A) durchd-Leu-l-Leu und in Circulin A durchd-Leu-Ll-Ile ersetzt. Im Polymyxin D₁ ist neben dem Ersatz der entsprechenden Sequenz durchl-Leu-l-Thr noch ein, -Diaminobuttersäurerest der Seitenkette durch eind-Serin ausgetauscht. Die entsprechenden Verbindungen mit dem Index 2 unterscheiden sich von denjenigen mit dem Index 1 durch einen Austausch der (+)-6-Methyloctansäure durch 6-Methylheptansäure.Die Struktur von Polymyxin B₁ E₁ und Circulin A konnte durch Totalsynthese gesichert werden (K.Vogler). Weitere Fortschritte in der Erforschung der Natur der noch unbekannten Vertreter sind in Kürze zu erwarten.     

Structure of albizziine [L(-)-2-amino-3-ureidopropionic acid], an amino acid from higher plants(Mimosaceae)

Zusammenfassung Eine neue Aminosäure, Albizziin, wurde durch chemische Verknüpfung zwischen N-Benzoylalbizziinmethylester undl-2-Benzamido-3-ureidopropionsäuremethylester (III) alsl-2-Amino-3-ureidopropionsäure (I) identifiziert. Die Synthese vonl-2-Ureido-3-aminopropionsäure (II) wird beschrieben.     

Catalytic properties of synthetic pentapeptides

Zusammenfassung Es wurde das Pentapeptidl-Thr-l-Ala-l-Abu-l-His-l-Asp synthetisiert, seine katalytische-hydrolytische Wirkung auf Essigsäure-p-nitrophenylester geprÜft und die katalytische Aktivität ca. 1/3 derjenigen seines isosterenl-Thr-l-Ala-l-Cys-l-His-l-Asp festgestellt.

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Work done on a leave of absence from the Research Council of Alberta, Canada.     

Rauwolfia alkaloids,XVI. Deserpidine,a new alkaloid fromRauwolfia canescens

Zusammenfassung AusRauwolfia canescens wurde Deserpidin, C₃₂H₃₈O₈N₂, isoliert und eine Konstitutionsformel für dieses neue Alkaloid vorgeschlagen.

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Communication XV,C. F. Huebner et al., J.A.C.S. (in press).

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From lecture given at Columbia University, New York, January 12th, 1955.     

Ferrous complexes in the catalase reaction

Zusammenfassung Vor kurzem wurde vonNicholls derWestheimer-Mechanismus des H₂O₂-Zerfalls durch Katalase mit Argumenten einer unseres Erachtens abwegigen Interpretation der polarographischen Kinetik kritisiert. Die Argumente werden durch Hinweise auf unberücksichtigte Arbeiten widerlegt.