Small bowel and colon glucose absorption study in rats by an adaptation of Sols and Ponz method.

An adaptation of the Sols and Ponz method for the study of glucose intestinal absorption was developed by considering the special conditions of our line research. The glucose absorption was studied in proximal jejunum, distal ileum and distal colon in Wistar rat. The main adaptations in the method for successive absorptions with intestinal perfusions in vivo were the length of the intestinal segment and the change of the pumping system. The results are very similar to those obtained with the original method.     

Identification of the Serratia marcescens hemolysin determinant by cloning into Escherichia coli.

A cosmid bank of Serratia marcescens was established from which DNA fragments were cloned into the plasmid pBR322, which conferred the chromosomally encoded hemolytic activity to Escherichia coli K-12. By transposon mutagenesis with Tn1000 and Tn5 IS50L::phoA (TnphoA), the coding region was assigned to a DNA fragment, designated hly, comprising approximately 7 kilobases. Two proteins with molecular weights of 61,000 (61K protein) and 160,000 (160K protein) were expressed by the pBR322 derivatives and by a plasmid which contained the hly genes under the control of a phage T7 promoter and the T7 RNA polymerase. When strongly overexpressed the 160K protein was released by E. coli cells into the extracellular medium concomitant with hemolytic activity. The genes encoding the 61K and the 160K proteins were transcribed in the same direction. Mutants expressing a 160K protein truncated at the carboxy-terminal end were partially hemolytic. Hemolysis was progressively inhibited by saccharides with increasing molecular weights from maltotriose (Mr 504) to maltoheptaose (Mr 1,152) and was totally abolished by dextran 4 (Mr 4,000). This result and the observed influx of [14C]sucrose into erythrocytes in the presence of hemolytic E. coli transformants under osmotically protective conditions suggest the formation of defined transmembrane channels by the hemolysin.     

Topology of proteolipid protein in the myelin membrane of central nervous system. A study using antipeptide antibodies

Peptides according to amino-acid sequences of the N- and C-terminus of lipophilin (proteolipid protein, PLP) (Gly1-Phe15 = 1; Thr261-Phe276 = 6) and of the other four hydrophilic domains (Glu37-Leu60 = 2; Arg97-Leu112 = 3; Gly119-Gly127 = 3A; Trp144-Tyr156 = 3B; Lys191-Ala203 = 4; Asn222-Phe232 = 5) have been synthesized by the solid-phase Fmoc method, linked covalently to keyhole limpet hemocyanin (KLH) and used as antigens. Monospecific antibodies against these antigens were isolated by affinity chromatography. Each antibody recognized its epitope in isolated partially delipidated PLP with the ELISA technique, western blot, thin sections of paraffin embedded rat brains and in the plasma membrane of appropriately fixed/permeabilized rat oligodendrocytes in culture. After fixation with formaldehyde antipeptide 3A antibody stained intact non-permeabilized cells. Therefore the epitope 3A must be located on the extracellular surface of the membrane. This is in full support of our previous biochemical results on the orientation of lipophilin in the myelin membrane.     

Reactions of sugar nitro-alkenes with acetoacetic esters

3,4,5,6,7-Penta-O-acetyl-1,2-dideoxy-1-nitro-d-gluco-hept-1-enitol reacts with methyl acetoacetate in an unusual Michael reaction, giving the normal adduct (6), and a bicyclic derivative (9) that arises from quasi-dimerization of the former when a high concentration of the base is used. Acetylation of compound 9 gives the hydroxylamine O-acetate (10).From the reactions of 3,4,5,6,7-penta-O-acetyl-1,2-dideoxy-1-nitro-d-galacto-hept-1-enitol with ethyl and tert-butyl acetoacetate, the normal adducts (7 and 8) were isolated. The structures of compounds 6 to 10 were established on the basis of their spectral properties (u.v., i.r., mass, and ¹H- and ¹³C-n.m.r.)     

Production of a potentially novel anti-microbial substance by Bacillus polymyxa

A bacterial strain, SCE2, identified as Bacillus polymyxa, produced an anti-microbial substance active against yeasts, fungi and different genera of Gram-positive and-negative bacteria, in liquid medium and in plate assays. This substance appeared to be an antibiotic different from the polymyxin group, mainly because of its action against the majority of Gram-positive bacteria tested and its lack of activity against Pseudomonas aeruginosa, a species usually killed by polymyxins. Preliminary characterization showed resistance to heat (65°C, 2 h), to proteases, trypsin, lysozyme, deoxyribonuclease I, ribonuclease A, phospholipase C, ethanol, acetone, chloroform, ether and to strong alkali treatment (2 M NaOH). The molecular weight was less than 3500. The B. polymyxa strain harboured a plasmid that did not correlate with antibiotic production; after curing experiments, a derivative strain, SCE2(46), was isolated that lacked the plasmid pES1, but showed the same inhibitory spectrum as the wild-type strain.     

Die optische Richtungsorientierung der Roten Waldameise (Formica Ruea L.)

Zusammenfassung Alle folgenden Angaben beziehen sich auf Formica rufa L., die Rote Waldameise, und sind nur unter Vorbehalt auf andere Insektenarten übertragbar.Die Ameisen benützen zur optischen Richtungsorientierung künstliche Lichtquellen, die Sonne oder den Mond.Eine distinkte Lichtquelle kann als Orientierungsmarke durch einen diffusen Lichtschein ersetzt werden.Mit Hilfe einer Polarisationsfolie läßt sich nachweisen, daß sich die Ameisen sowohl nach der Schwingungsrichtung des blauen Himmelslichtes als auch nach der Schwingungsrichtung des Folienlichtes orientieren können.Die Orientierung nach Landmarken, wie Häusern und Bäumen, spielt eine große Rolle und ist bei bewölktem Himmel wahrscheinlich die einzige optische Orientierungsmöglichkeit.Werden Himmels- und Landmarken in Konkurrenz gesetzt, dann läuft die Ameise in einer Kompromißrichtung.Ameisen reagieren in Neststimmung vorwiegend negativ und in Exkursionsstimmung vorwiegend positiv phototaktisch.Es wird eine Methode angegeben, mit der durch Vergleich von Dreherregungen die Stärke der phototaktischen Drehreaktionen gemessen werden kann.Bei gleich großer Ablenkung vom orientierten Lauf sind die geotaktischen und die phototaktischen Dreherregungen (Drehtendenzen) quantitativ gleich.Die phototaktischen Dreherregungen (Drehtendenzen) sind helligkeitsunabhängig, ändern sich jedoch mit dem Einfallswinkel des Lichtes.Aus den experimentellen Befunden wird geschlossen, daß sich am zentralnervösen Funktionsgefüge der negativen (positiven) Phototaxis mindestens drei nervöse Instanzen (Mechanismen) beteiligen: Das Integrationszentrum, das Lagezentrum und der Koordinationsmechanismus der Beinbewegung.Wichtige Vorgänge beim Übergang von der positiven und negativen Phototaxis zur menotaktischen Hin- und Rückwegorientierung sind orientierungsfreie Suchschleifen und Lernprozesse, die zur Ermittlung der Luftlinienrichtung führen.Diese Lernprozesse finden sowohl auf dem Hin- als auch auf dem Rückweg statt.Die Ermittlung der Luftlinienrichtung geschieht über die Auswertung (Integration) der optischen Reizfolge, die kinästhetische Reizfolge ist dafür wahrscheinlich völlig bedeutungslos.Mit Hilfe der Kompensationstheorie werden eine Reihe von Reaktionen sich menotaktisch orientierender Ameisen kausal erklärt.Die Ameise kann sich eine Laufrichtung in bezug auf eine Lichtquelle mindestens 5 Tage lang merken.Die Ameise kann sich mit Hilfe von Landmarken an mindestens vier verschiedenen Plätzen im Gelände je eine bestimmte Laufrichtung merken.Erinnerungsbilder von Himmels- und Landmarken werden im Gedächtnis der Ameisen unabhängig voneinander aufbewahrt, die Erinnerungsbilder der Himmelsmarken dagegen sind im Gedächtnis der Ameisen aneinandergekoppelt.Die Ameisen haben die Fähigkeit, die Wanderung der Sonne bei der Richtungsorientierung mit einzuberechnen.     

Arabidopsis ADC1 functions as an N~δ-acetylornithine decarboxylase

Polyamines are small aliphatic amines found in almost all organisms, ranging from bacteria to plants and animals. In most plants, putrescine, the metabolic precursor for longer polyamines, such as spermidine and spermine, is produced from arginine, with either agmatine or ornithine as intermediates. Here we show that Arabidopsis thaliana(Arabidopsis) arginine decarboxylase 1(ADC1), one of the two known arginine decarboxylases in Arabidopsis, not only synthesizes agmatine from arginine, but also converts N~δ-acetylornithine to N-acetylputrescine. Phylogenetic analyses indicate that duplication and neofunctionalization of ADC1 and NATA1, the enzymes that synthesize N~δ-acetylornithine in Arabidopsis, co-occur in a small number of related species in the Brassicaceae. Unlike ADC2, which is localized in the chloroplasts, ADC1 is in the endoplasmic reticulum together with NATA1, an indication that these two enzymes have access to the same substrate pool. Together, these results are consistent with a model whereby NATA1 and ADC1 together provide a pathway for the synthesis of N-acetylputrescine in Arabidopsis.