Nuclear Overhauser effect investigation on GM1 ganglioside containingN-glycolyl-neuraminic acid (II3Neu5GcGgOse4Cer)

The conformational properties of the oligosaccharide chain of GM1 ganglioside containingN-glycolyl-neuraminic acid, -Gal-(1-3)--GalNAc-(1-4)-[-Neu5Gc-(2-3)]--Gal-(1-4)--Glc-(1-1)-Cer, were studied through NMR nuclear Overhauser effect investigations on the monomeric ganglioside in dimethylsulfoxide, and on mixed micelles of ganglioside and dodecylphosphocholine in water. Several interresidual contacts for the trisaccharide core--GalNAc-(1-4)-[-Neu5Gc-(2-3)]--Gal-were found to fix the relative orientitation of the three saccharides, while the glycosidic linkage of the terminal -Gal-was found to be quite mobile as the -Gal-(1-3)--GalNAc-disaccharide exists in different conformations. These results are similar to those found for two GM1 gangliosides containingN-acetyl-neuraminic acid and neuraminic acid [1].Abbreviations Ganglioside nomenclature is in accordance with Svennerholm [23] and the IUPAC-IUB Recommendations [24] GM3(Neu5Ac) II³Neu5AcLacCer, -Neu5Ac-(2-3)--Gal-(1-4)--Glc-(1-1)-Cer - GM3(Neu5Gc) II³Neu5GcLacCer, -Neu5Gc-(2-3)--Gal-(1-4)--Glc-(1-1)-Cer - GM1(Neu5Ac) II³Neu5AcGgOse₄Cer, -Gal-(1-3)--GalNAc-(1-4)-[-Neu5Ac-(2-3)]--Gal-(1-4)--Glc-(1-1)-Cer - GM1(Neu5Gc) II³Neu5GcGgOse₄Cer, -Gal-(1-3)--GalNAc-(1-4)-[-Neu5Gc-(2-3)]--Gal-(1-4)--Glc-(1-1)-Cer - GM1(Neu) II³NeuGgOse₄Cer, -Gal-(1-3)--GalNAc-(1-4)-[-Neu-(2-3)]--Glc-(1-1)-Cer - GD1a IV³Neu5AcII³Neu5AcGgOse₄Cer, -Neu5Ac-(2-3)--Gal-(1-3)--GalNAc-(1-4)-[-Neu5Ac-(2-3)]--Gal-(1-4)--Glc-(1-1)-Cer - GalNAc-GD1a IV⁴GalNAcIV³Neu5AcII³Neu5AcGgOse₄Cer, -GalNAc-(1-4)-[-Neu5Ac-(2-3)]--Gal-(1-3)--GalNAc-(1-4)-[-Neu5Ac-(2-3)]--Gal-(1-4)--Glc-(1-1)-Cer - Neu neuraminic acid - Neu5Ac N-acetyl-neuraminic acid - Neu5Gc N-glycolyl-neuraminic acid - Cer ceramide     

Genetic characterization of a new splice variant of the β2 subunit of the voltage-dependent calcium channel

This study reports a novel splice variant form of the voltage-dependent calcium channel ₂ subunit (_2g). This variant is composed of the conserved amino-terminal sequences of the _2a subunit, but lacks the -subunit interaction domain (BID), which is thought essential for interactions with the ₁ subunit. Gene structure analysis revealed that this gene was composed of 13 translated exons spread over 107 kb of the genome. The gene structure of the ₂ subunit was similar in exon-intron organization to the murine ₃ and human ₄ subunits. Electrophysiological evaluation revealed that _2a and _2g affected channel properties in different ways. The _2a subunit increased the peak amplitude, but failed to increase channel inactivation, while _2g had no significant effects on either the peak current amplitude or channel inactivation. Other subunits, such as ₃ and ₄, significantly increased the peak current and accelerated current inactivation.     

Cluster Maintenance in Mobile Ad-hoc Networks

The main contribution of this paper is to propose a new cluster maintenance algorithm and a companion cluster initialization algorithm based on a number of interesting and novel properties of diameter-2 graphs. The initialization algorithm naturally blends into cluster maintenance, showing the unity between these two operations. We refer to our algorithms as tree-based since they depend on a spanning tree maintained at various nodes. Unlike the vast majority of published clustering algorithms, our algorithms are cluster-centric, as opposed to node-centric, and work in the presence of node mobility. Extensive simulation results have shown the effectiveness of our algorithms when compared to other clustering schemes proposed in the literature.Lan Wang received his B.S. and M.S. degrees in computer science from Harbin Engineering University, China in 1992 and 1995, respectively. From 1995 to 1999, he worked as a software engineer at the System Engineering Research Institute of CSSC, Beijing, China. He is currently a PhD student at the Computer Science Department of Old Dominion University.Stephan Olariu received the M.Sc. and Ph.D. degrees in computer science from McGill University, Montreal, Canada in 1983 and 1986, respectively. In 1986 he joined the Computer Science Department at Old Dominion University where he is now a full professor. He has published extensively in various journals, book chapters, and conference proceedings. His research interests include wireless networks and mobile computing, parallel and distributed systems, performance evaluation, and medical image processing. Prof. Olariu serves on the editorial board of several archival journals including IEEE Transactions on Parallel and Distributed Systems, Journal of Parallel and Distributed Computing, International Journal of Foundations of Computer Science, Journal of Supercomputing, International Journal of Computer Mathematics, VLSI Design, and Parallel Algorithms and Applications.     

Expression of a chimeric stilbene synthase gene in transgenic wheat lines

A chimeric stilbene synthase (sts)gene was transferred into wheat. Stilbene synthases play a role in the defence against fungal diseases in some plant species (e.g. groundnut or grapevine) by producing stilbenetype phytoalexins like resveratrol. Resveratrol is also claimed to have positive effects to human health. Embryogenic scutellar calli derived from immature embryos of the two commercial German spring wheat cultivars Combi and Hanno were used as target tissue for cotransformation by microprojectile delivery. The selectable marker/reporter gene constructs contained the bargene either driven by the ubiquitinpromoter from maize (pAHC 25, also containing the uidAgene driven by the ubiquitinpromoter), or by the actinpromoter (pDM 302) from rice. The cotransferred plasmid pStil 2 consisted of a grapevine stscoding region driven by the ubiquitin promoter. Eight transgenic Combi and one Hanno T_Oplant were obtained and, except one Combi T_Oplant, found to be cotransformants due to the integration of both the stsgene and the selectable marker or reporter genes. Expression of the stsgene was proven by RTPCR, and, for the first time, by detection of the stilbene synthase product resveratrol by HPLC and mass spectrometry. The stsgene was expressed in four of the seven transgenic Combi T_{_o}plants. Two of the respective T₁progenies segregated in a Mendelian manner were still expressing the gene. Investigations into methylation of the stsgene showed that in three nonexpressing progenies inactivation was paralleled by methylation.     

Agonistisches Verhalten, Territorialverhalten und Balz der Zwergtrappe (Tetrax tetrax)

Zusammenfassung 1980–1984 wurden in Südfrankreich und Portugal Untersuchungen zum agonistischen Verhalten, Territorialverhalten und zur Balz der ZwergtrappeTetrax tetrax durchgeführt.Während der Fortpflanzungsperiode kommt es zwischen Zwergtrappen- häufig zu agonistischen Auseinandersetzungen. Ausdruck aggressiver Erregung ist enges Anlegen des Halsgefieders, Langstrecken des Halses, und nur beim Aufrichten und dachartiges Falten der Schwanzfedern. Aggressive Verhaltensweisen sind Drohen durch Hinlaufen oder -fliegen zu Reviereindringlingen, seitliches Drohen gleichstarker , und nur, wenn Verhaltensweisen niedrigerer Intensität nicht zur Unterlegenheit eines der Rivalen führen, Schnabelkämpfe.Rufen dient der Markierung eines Reviers. Die Intensität des Rufens wird durch die Anwesenheit von nicht beeinflußt, die Anwesenheit fremder nahe der Reviergrenze führt jedoch zu einem deutlichen Anstieg. Die Rufbewegung macht eine gewisse optische Signalwirkung, vor allem auf kurze Distanz, zusätzlich zur akustischen Wirkung, wahrscheinlich.Fliegende erzeugen mit den Schwingen (besondere Struktur der 7. Handschwinge) ein pfeifendes Geräusch, das während der Fortpflanzungszeit Bedeutung in der innerartlichen Kommunikation hat. Es zeigt sowohl als auch den Anflug eines weiteren an und löst damit territoriale bzw. aggressive Verhaltensweisen oder Flucht aus. Weitere Bedeutung erlangt es als Element des Territorialen Flügelschlagens und der Sprungbalz.Über ihrem Revier fliegen territoriale stets mit leicht hochgebogenem Kopf und verhaltenen, flachen Flügelschlägen. Deutungen dieses Fluges als Imponierflug zur besseren Darstellung des auffällig gefärbten Halsgefieders bzw. als Suchflug zum leichteren Auffinden und Verjagen von Reviereindringlingen werden diskutiert.Territoriales Flügelschlagen beginnt mit Fußtrampeln, das sich beschleunigt und in einen Ruf überleitet. Gleichzeitig schlägt das dreimal sehr schnell und flach mit den Flügeln, hebt jedoch nicht vom Boden ab. Alle Elemente des Verhaltens sind deutlich zu hören. Der Verstärkung der beim Fußtrampeln erzeugten Klopfgeräusche dienen Balzplätze, die entweder auf akustisch besonders geeignetem Boden angelegt oder durch das Fußtrampeln der sekundär verbessert werden.Territoriales Flügelschlagen wird ausschließlich in niedrigen Lichtintensitäten während kurzer Zeit in der Morgen- und Abenddämmerung gezeigt. Die Anwesenheit von hat keinen Einfluß auf seine Intensität. Es ist eine territoriale Verhaltensweise mit akustischem Signal und wird als ritualisiertes Anlaufen gegen einen Reviereindringling bzw. ritualisiertes Auffliegen eines Revier- zum Eindringling hin gedeutet. Optische Komponenten kommen in der deckenden Vegetation kaum zur Geltung. können wegen der relativ geringen Reichweite der Signale nicht angelockt werden.Sprungbalz tritt zeitlich streng getrennt vom Territorialen Flügelschlagen in wesentlich höheren Lichtintensitäten auf; seine Intensität hängt ab von Kontakten zu . Sie ähnelt zwar in der Ausführung dem Territorialen Flügelschlagen, der Vogel hebt sich jedoch während der langsameren Flügelschläge durch einen Sprung vom Boden ab, und das Fußtrampeln ist wesentlich weniger intensiv. Charakteristische Flügelbewegungen während des Balzsprunges exponieren schwarzweiße Gefiederpartien bis in 65 cm Höhe. Sprungbalz erhöht im Vergleich zu Territorialem Flügelschlagen stark die optische Auffälligkeit eines ; die Sprunghöhe garantiert zusammen mit der Lage der Balzplätze bei geringstmöglichem Energieaufwand eine maximale Sichtbarkeit des über der umgebenden Vegetation. Sowohl die zeitliche Korrelation der Sprungbalz mit den Aktivitäten der als auch die Art der Interaktionen mit während Sprungbalzphasen machen deutlich, daß diese Verhaltensweise ins Paarungsrevier zieht.Das Hennenjagen dient der Stimulierung der zur Kopulation. Das läuft in charakteristischer Körperhaltung schnell hinter einem her, hat dabei den Kopf tief in die aufgerichtete Halskrause eingezogen, hält oft ruckartig an und ruft unter Zurseitewerfen des Kopfes. Kopulationen sind sehr kurz. Offensichtlich kann das Hennenjagen die sonst sehr ausgeprägte Abwehr des gegen das unterdrücken und zur Kopulationsbereitschaft führen. Die Kopulation selbst jedoch hebt diesen Effekt wieder auf, so daß nach 1–2 Sekunden das abwehrt oder flieht.

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Agonistic behaviour, territorial behaviour and courtship display of the Little Bustard (Tetrax tetrax)

Summary From 1980 to 1984 studies were made of the agonistic, territorial and courtship behaviour of the Little Bustard in Southern France and Portugal. The behaviour patterns are described in detail, the signals involved are analysed, and factors which could have led to their evolution are discussed.In the reproductive period agonistic encounters between males happen remarkably often. Aggressive excitement is expressed in both sexes by compression of the neck-feathers and stretching of the neck, and in the female by the erection and lateral folding of the tail-feathers. There are several kinds of threat behaviour in territorial encounters: running or flying towards intruders; lateral threat behaviour (if both males are of similar strength); and bill-fighting which only occurs if aggressive behaviour of a lower intensity has not led to the submission of one of the rivals. Calling is a territorial behaviour and serves to mark a territory. The frequency of calls is not affected by the presence of females, but the presence of other males close to the territory borders can result in a marked increase in calling frequency. In addition to the acoustic signal, the call involves a sharp neck-jerk which may act as an optical signal, at least over short distances.In flight males produce a whistling sound by means of the specially adapted 7th primary feather on their wings. This sound has important meaning in intraspecific communication during the reproductive period: it announces a flying male to other males as well as to females, and causes territorial and aggressive behaviour, or fleeing of these birds. The flight sound is also an important element of the behaviour patterns territorial wingbeat display and jumping display. Within their territories males always fly with the neck raised at a slight angle and with suppressed, shallow wing-beats. Two interpretations of this flight are discussed: that it serves to advertise the conspicuously coloured neck-feathers in a display; or that it is a search flight for locating and chasing-off intruders.Territorial wingbeat display begins with an accelerating foot-stamping and leads into a call. During the call the male performs three very fast, whistling wing-beats but remains on the ground. The combined elements, foot-stamping, calling and whistling wings, produce a unique and distinctive acoustic signal. The foot-stamping is only performed at special display sites where the sound is amplified by the substrate structure. At such sites, the soil typically has trapped pockets of air below a compacted surface which may result from the repeated defecating and stamping of the male on the same spot. Territorial wingbeat display behaviour is only performed for short periods in low light intensities at dawn and dusk. The presence of females has no effect on the intensity of the behaviour which serves a territorial function. It is interpreted to be a ritualisation of the aggressive running or flying of a territorial male towards an intruder. Optical elements of this behaviour cannot have much importance because the body and wings of the bird are rarely visible above the vegetation. Territorial wingbeat display behaviour cannot be seen over long distances and from this reason cannot serve to attract females into a males territory.Jumping display is only performed at much higher light intensities than territorial wingbeat display so the two never occur at the same time of day. The intensity of the behaviour increases markedly in the presence of females in sharp contrast to the territorial wingbeat display. The jumping display is performed in a similar way to the territorial wingbeat display except that the foot-stamping is much less intensive, the wing-beats are slower, and the bird jumps off the ground during the wing-beats. During the jumping display the black and white patterns on the body and wings are clearly visible and the behaviour increases the conspicuousness of the male markedly. The jump, advertising the wing-pattern up to a height of 65 cm, together with the specific location of the display site, ensures that maximum visibility of the male above the vegetation is achieved at minimum energetic cost. Activities of the females and their interactions with males during the jumping periods indicate that this behaviour serves to attract females to the males territory.The chasing of females is also a courtship behaviour and serves in stimulating and preparing females for copulation. In a characteristic posture with the head retracted into the neck-collar, the male rapidly runs behind the female, repeatedly stopping abruptly and calling whilst throwing its head and body sideways. Copulations are performed very quickly and only happen after a female has been chased for some time. Under certain preconditions chasing suppresses the aggressive and defence behaviour of the female which normally characterises encounters with males, and thus leads to readiness for copulation. Copulation itself removes this effect and after only one or two seconds aggression leads to the escape of the female.

     

Polymorphism of the HLA DR1 haplotype in the Israeli population investigated at the serological,cellular, and genomic levels

In the present report, we used serological, cellular, and restriction fragment length polymorphism (RFLP) to investigate the DR1 haplotype in the Israeli population. We describe an Israeli homozygous typing cell (HTC), HLA-DwLVA, which defines a new lymphocyte-activating determinant associated with Bw65, DR1 and distinct from Dwl. The parents of this donor, non-Ashkenazi Algerian Jews, are first cousins and share HLA-Cw8, Bw65, BfS, DR1, DQw1, DPw4. No specificity could be assigned to HLA-DwLVA using the 91 Ninth Workshop HTCs. Two families and forty unrelated DR1 individuals were studied with DwLVA and a panel of DR1/Dw1 HTCs. HLA-DwLVA showed segregation as a single determinant within families. This new specificity was present in 24 out of 40 (60%) unrelated DR1 individuals, indicating that in the Israeli population DwLVA is the main lymphocyte-defined determinant associated with the serologically defined DRI specificity, in contrast to non-Jewish Caucasoids where DR1 is significantly associated with Dw1. The vast majority of DwLVA-positive carriers were also Bw65 carriers, indicating that Bw65, DR1, DwLVA may represent a typical allele combination in the Israeli population. The RFLP analysis established the correlation of certain RFLPs with Dw1 and DwLVA. In addition, we describe a cluster of RFLPs that may correspond to a new Dw subtype associated with DR1, for which no serological and cellular reagents have been described so far.     

Identification and DNA sequence analysis of a γ-type gliadin cDNA plasmid from winter wheat

Summary Recombinant cDNA plasmids possessing the coding sequences for the -type gliadins were isolated from a cDNA library prepared from wheat seed poly (A⁺) RNA. One of these plasmids, pGliB48, specifically hybridizes to poly (A⁺) RNA molecules 1 400–1 500 bases in length that direct the synthesis of polypeptides at 38 Kd and 46 Kd, the latter size characteristic of the -type gliadins. The cDNA sequence of pGliB48 was determined and encompasses the 3 untranslated region as well as 245 amino acids from the C-terminus of the -type gliadin polypeptide. The 5-end of the DNA coding sequence consists of a tandem repeat unit composed of eight amino acids. Localized regions of homology are observed for the /-type and -type gliadin cDNA sequences.     

Charles Darwin's theory of evolution: A review of our present understanding

The paper characterizes Darwin's theory, providing a synthesis of recent historical investigations in this area. Darwin's reading of Malthus led him to appreciate the importance of population pressures, and subsequently of natural selection, with the help of the wedge metaphor. But, in itself, natural selection did not furnish an adequate account of the origin of species, for which a principle of divergence was needed. Initially, Darwin attributed this to geographical isolation, but later, following his work on barnacles which underscored the significance of variation, and arising from his work on botanical arithmetic, he supposed that diversity allowed more places to be occupied in a given region. So isolation was not regarded as essential. Large regions with intense competition, and with ample variation spread by blending, would facilitate speciation. The notion of place was different from niche, and it is questioned whether Darwin's views on ecology were as modern as is commonly supposed. Two notions of struggle are found in Darwin's theory; and three notions of variation. Criticisms of his theory led him to emphasize the importance of variation over a range of forms. Hence the theory was populational rather than typological. The theory required a Lamarckian notion of inheritable changes initiated by the environment as a source of variation. Also, Darwin deployed a use/habit theory; and the notion of sexual selection. Selection normally acted at the level of the individual, though kin selection was possible. Group selection was hinted at for man. Darwin's thinking (and also the exposition of his theory) was generally guided by the domestic-organism analogy, which satisfied his methodological requirement of a vera causa principle.     

Excitatoren und paarungsverhalten mitteleuropäischer malachiiden (Coleopt., Malacodermata)

Ohne ZusammenfassungAlphabetisches Verzeichnis der im Text gebranchten Abkürzungen AK Anbieten der Kopfgrube () - AR-seitig außenrandseitig (auf Elytre bezogen) - EO Elytralorgan (EO-Arten = Arten mit Elytralorganen im männlichen Geschlecht) - f Flucht () - F Flucht () - FA, fa frontale Auseinandersetzung (, ) - FS, fs Fühlertrillern bzw. Frontalspiel (, ) - gk Grubenknabbern () - IR-seitig innenrandseitig (auf Elytre bezogen) - K Kopulation - KG Kopfgrube (KG-Arten = Arten mit Kopfgrube im männlichen Geschlecht) - KI Abdomenkitzeln () - KV Kopulationsversuch () - LP-Feld den weiblichen Labialpalpen korreliertes Drüsenporenfeld - MP-Feld den weiblichen Maxillarpalpen korreliertes Drüsenporenfeld - 180 Drehung des um 180° - ob Organbeißen () - ok Organknabbern () - OZ Organzuwendung () - P Prüfung der Kopulationsbereitschaft durch - RB rückwärtige Berührung durch - SLV Seitwärtslauf nach vorn () - SLH Seitwärtslauf nach hinten () - U Umrundung () - vl Vorwärtslauf () - 180 Drehung des um 180° Habilitationsschrift.     

Saccharomyces cerevisiae 2-mum DNA. An analysis of the monomer and its multimers by electron microscopy.

Summary The non-tandem inverted duplication in the 2-m DNA of Saccharomyces cerevisiae has a length of 0.19 m and is located asymmetrically along the molecule. The majority of the dumb-bell structures that are formed upon denaturation and selfannealing of the 2-m monomer consists of the renatured inverted duplication sequences as double stranded stem and two single stranded loops of 0.67 m±0.06 m (S-loop) and 0.86 m±0.05 m (L-loop) length. Two additional size classes which comprised 5–10% of the measured molecules had contour lengths of around 1.7 m and 2.1 m. The smaller dumb-bells contained two S-loops and the larger dumb-bells contained two L-loops as was shown by heteroduplex mapping with an HindIII fragment from the L-loop. Two models which assume illegitimate or site specific recombination, are presented to explain the generation of double S-loop and double L-loop molecules. At least part of the 4-m and 6- circular molecules present in the yeast supercoiled DNA fraction are shown to be dimers and trimers of 2-m monomers, but often with inverted loop segments most probably due to intramolecular recombination between sequences of the inverted duplication.2-m DNA is used to indicate the supercoiled DNA fraction although in our measurements the average monomeric length is 1.9 mPart of this work has been presented at the Conference: The Genetics and Biogenesis of Chloroplasts and Mitochondria, Munich, August, 1976     

Occurrence of airborne Cladosporium and Alternaria spores in Southern and Central Poland in 1995–1996

The concentration of airborne spores of Cladosporium and Alternaria has been investigated at five monitoring stations situated in cities from the foot of the Tatra Mountains to central Poland along a south-north transect (Zakopane, Kraków, Ostrowiec witokrzyski, Warszawa, Pozna) i.e. from a height of 900 m to 80 m above sea level. The aerobiological monitoring of fungal spores was performed by means of five Burkard volumetric spore traps.Cladosporium spores were dominant at all the stations. The highest Cladosporium and Alternaria spore concentrations were observed at all the sites in July and August, except at Warszawa in both years and at Pozna in 1995 where the maximum of Cladosporium spores occurred in June and July, and at Ostrowiec witokrzyski in 1995 where the maximum was found in July, August and September.Fungal spore concentrations in Zakopane and Kraków were significantly lower than those in Ostrowiec witokrzyski, Warszawa and Pozna and periods of abundant Cladosporium spore occurrence were different in these two groups of monitoring stations.     

Modulation of natural killer activity by thymosin alpha 1 and interferon

Summary A single injection of -interferon (-IFN) (30 000 units/mouse), a major biological modifier of natural killer (NK) cytolytic activity, strongly stimulated NK activity in normal mice, as expected, while the same treatment did not statistically alter the NK response in cyclophosphamide (CY)-suppressed animals.We investigated the possibility of thymosin ₁ cooperating with -IFN in boosting NK activity in CY-suppressed animals.The results show that treatment with thymosin ₁ (200 g/kg) for 4 days, followed by a single injection of -IFN 24 h before testing, strongly restored NK activity in CY-suppressed mice. Thymosin ₁ was, moreover, able to accelerate the recovery rate of NK activity in bone marrow reconstituted murine chimeras.Taken together the data support the concept that the synergic effect between thymosin ₁ and -IFN could be the result of effects on differentiation of the NK lineage at different levels.     

In vitro meristem cloning of Eucalyptus tereticornis Sm.

Rapid multiplication of axillary meristems and direct shoot development occurred from nodal explants of mature Eucalyptus tereticornis Sm. with 5.3 M NAA, 1.1 M IAA and 4.4 M BA in Murashige-Skoog medium. Repeated subcultures of the second generation shoot cultures into low cytokinin-auxin containing media (0.44–0.88 M BA+0.1 M NAA) yielded axillary microshoots in large numbers. Half-strength MS liquid medium with 4.9 M IBA, 5.5 M IAA and 5.3 M NAA for four days, half-strength semi-solid hormonefree MS medium with charcoal, and MS liquid medium without charcoal and hormones, in sequence, induced rooting of shoots in the dark. This system is suitable for the mass propagation of this difficult-to-root eucalypt.Abbreviations BA 6-benzyladenine - IAA indole-3-acetic acid - IBA -indole-3-butyric acid - 2-iP isopentyl adenine - Kn kinetin - MS Murashige-Skoog - NAA -naphthalene acetic acid - PVP polyvinylpyrrolidone     

Primary structure of the hemoglobin beta-chain of rose-ringed parakeet (Psittacula krameri)

The primary structure of Rose-ringed Parakeet hemoglobin -chain was established, completing the analysis of this hemoglobin. Comparisons with other avian -chains show variations smaller than those for the corresponding -chains. There are 11 amino acid exchanges in relationship to the only other characterized psittaciform -chain, and a total of 35 positions are affected by differences among all avian -chains analyzed (versus 61 for the -chains). At three positions, the Psittacula -chain has residues unique to this species. Three ₁₁ contacts are modified, by substitutions at positions 51, 116, and 125.     

Sialidase of swine influenza A viruses: variation of the recognition specificities for sialyl linkages and for the molecular species of sialic acid with the year of isolation

The sialidase of swine influenza A viruses of N1 and N2 subtypes, isolated from 1930 to 1992, was studied for substrate specificity with ganglio-series, lacto-series type II and GM3 gangliosides containing Neu5Ac2-3Gal, Neu5Gc2-3Gal and Neu5Ac2-6Gal linkages. All viral sialidases tested showed that the activity for hydrolysing substrates with Neu5Ac2-3Gal was higher than the activities with Neu5Gc2-3Gal and Neu5Ac2-6Gal linkages. When GM1b, GM3 and sialylparagloboside were used as substrates, the earliest strain (A/Wisconsin/15/30 H1N1, isolated in 1930) showed the activity ratio of Neu5Ac2-6Gal to Neu5Ac2-3Gal to be 0.13:0.2, and the ratio Neu5Gc2-3Gal/Neu5Ac2-3Gal to be 0.19:0.37, while those strains isolated from 1978 to 1992 exhibited ratios of 0.29:0.58 for Neu5Ac2-6Gal/Neu5Ac2-3Gal and 0.51:0.76 for Neu5Gc2-3Gal/Neu5Ac2-3Gal. The above results indicate that the substrate specificities of sialidases from swine influenza A viruses towards sialyl linkages and the molecular species of sialic acid are related to the year of isolation, i.e. strains isolated after 1978 exhibited higher activity towards Neu5Ac2-6Gal and Neu5Gc2-3Gal linkages when compared with strains isolated in an earlier year, 1930.Abbreviation Neu5Ac 5-N-acetylneuraminic acid - Neu5Gc 5-N-glycolyneuraminic acid - Gal d-galactose - Glc d-glucose - Cer Ceramide - II³(Neu5Ac)Lac Neu5Ac2-3Gal1-4Glc - GM3(Neu5Ac2-3Gal) Neu5Ac2-3Gal1-4Glc1-Cer - GM3(Neu5Gc2-3Gal) Neu5Gc2-3Gal1-4Glc1-Cer - GM1b(Neu5Ac2-3Gal) Neu5Ac2-3Gal1-3GalNac1-4Gal1-4Glc1-Cer - GMlb(Neu5Gc2-3Gal) Neu5Gc2-3Gal1-3GalNAc1-4Gal1-4Glc1-Cer - IV³(Neu5Ac)nLc4Cer Neu5Ac2-3Gal1-3GlcNAc1-4Gal1-4Glc1-Cer - IV³(Neu5Gc)nLc4Cer Neu5Gc2-3Gal1-3GlcNAc1-4Gal1-4Glc1-Cer - IV⁶(Neu5Ac)nLc4Cer Neu5Ac2-6Gal1-3GlcNAc1-4Gal1-4Glc1-Cer - TDC taurodeoxycholate.     

One-bond carbon-proton coupling constants: Angular dependence in β-linked oligosaccharides

Summary The angular dependence of¹J_C,H in model compounds related to -linked oligosaccharides has been established by FPT INDO quantum chemical calculations. Values calculated for models of (1 1)-, (1 2)-, (1 3)- and (1 4)-linked disaccharides were compared, and the effect of the orientation of HO-2 elucidated. The angular dependence of¹J_C,H on the torsional angles ^H and ^H and the solvent dielectric constant (s) was characterized in the form:¹J_C,H = A cos2+B cos + C sin2 + D since + E + Fe. The¹J_C,H values, measured by DEPT methods for C-1-H-1 and C-X-H-X in cellobiose, cyclic trisaccharide and hexopyranoses were used to adjust the calculated angular dependences. Based on the occurrence of the conformers for agarobiose, neoagarobiose, mannobiose and methyl -xylobioside, the thermodynamically averaged <¹J_C,H > values were calculated. The results obtained (<¹J_C-1,H-1 > 162.4, <¹J_{C-4, H-4} > 147.6 Hz for methyl -xylobioside; <¹J_C-1,H-1 > 162.4 and <¹J_C-4,H-4] > 147.6 Hz for mannobiose; <¹J_C-1,H-1 > 162.8 Hz for neo agarobiose and <¹J_C-1,H-1 > 163.2 Hz for agarobiose) agree well with the experimental values of 162.7, 147.5, 160.4, 147.2, 160.9 and 165.7 Hz, respectively.     

Design of superactive and selective integrin receptor antagonists containing the RGD sequence

Summary Integrins play a major role in cell-cell and cell-matrix interactions. The majority of the different types of integrins recognize the tripeptide sequence arginine-glycine-aspartic acid (RGD). To explore the spatial requirements of the pharmacophore for receptor selectivity and high activity, a new procedure, spatial screening, was used. The procedure is based on the experience that the conformation of small cyclic peptides is mainly determined by the chirality of the amino acids (and glycine or proline). For example, cyclic pentapeptides with one d and four l amino acids prefer a II'/ conformation. The sequence RGDFV was shifted around this spatial II'/ template by synthesis of five peptides in which one of the amino acids was used in d-configuration. It turned out that cyclo(-RGDfV-) is a selective inhibitor for the _v3 integrin, which is strongly expressed in cancer cells. Systematic variations with different turn mimetics, retro-inverso structures, modified peptide bonds and sugar amino acids are discussed.     

A simple brassinolide analogue 2α, 3α-dihydroxy-17β-(3-methyl-butyryloxy)-7-oxa-B-homo-5α-androstan-6-one which induces bean second internode splitting

A new synthetic brassinolide analogue, 2,3-dihydroxy-17-(3-methylbutyryloxy)-7-oxa-B-homo-5-androstan-6-one (11), has been shown to exhibit typical brassinolide activity characterised by elongation, swelling, twisting and splitting of the bean second internode. It was prepared from the known lactone 2,3,17-trihydroxy-7-oxa-B-homo-5-androstan-6-one (4) which was transformed to an isopropylidenedioxy derivative. After protection of the 2- and 3-hydroxy groups it yielded the 2,3-isopropylidenedioxy-17-(3-methyl-butyryloxy)-7-oxa-B-homo-5-androstan-6-one (7) on treating with 3-methylbutyryl chloride in pyridine. The analogue with a 2-methylbutyric moiety (10, 2,3-dihydroxy-17-(2-methyl-butyryloxy)-7-oxa-B-homo-5-androstan-6-one) in position 17 stimulated only elongation and swelling of the bean second internode. However, in this bioassay 100 times more 10 or 11 compared to 24-epibrassinolide is required to obtain the same effects. Analogues with -oriented hydroxyl groups at C-2 and C-3 (14,15), a 6-ketone (17,18) or 6-oxa-7-oxo-lactone system (12,13) in ring B lack the typical brassinolide activity. In addition, the active brassinosteroids applied to the second internode stimulated a similar, but 30% lower elongation of the first internode. From data presented here we conclude that the presence of two hydroxy groups in the positions 22 and 23 of the brassinolide side chain, which are considered as a key structural requirement, is not absolutely necessary for a compound to exhibit typical brassinosteroid activity. Nevertheless, these compounds have generally 2–10 times lower activity than that having 22,23-vicinal diol in the side chain.     

Distribution of pectic polysaccharides throughout walls of suspension-cultured carrot cells

Summary A monoclonal antibody (2 F 4) recognizing a conformational epitope of polygalacturonic acid was used for immunogold localization of pectins in walls of suspension-cultured carrot (D. carota L.) cells at the electron microscopic level. In microcolonies of young or mature cells, polygalacturonic acid was essentially located on the middle lamella material expanded at three-way junctions between cells or lining intercellular spaces but was not found in primary walls. Middle lamellae far from junction zones and intercellular spaces were not recognized. Largely esterified pectic polymers, only detected by the 2 F 4 antibodies after on-grid de-esterification treatment by pectin methyl esterases, were present within all primary cell walls. Golgi bodies and associated vesicles were also labeled by the 2 F 4 antibodies only after de-esterification treatment, which indicates that pectic polymers are synthesized and secreted in a highly esterified form. A decrease of pectin esterification, which results probably from an in situ enzymatic de-esterification of the pectic polymers of the primary walls, was observed in senescent cells. These results are discussed in relation to biochemical analyses showing changes of the methyl ester content of pectins during the cell-wall growth.