Carbohydrates of influenza virus. Structure of the oligosaccharides linked to asparagines 406 and 478 in the hemagglutinin of fowl plague virus,strain dutch

Fowl plague virus, strain Dutch, was metabolically labeled withd-[2-³H]mannose, or withd-[6-³H]glucosamine, and the small subunit (HA₂; 0.8 mg in total) of the viral hemagglutinin was isolated by preparative sodium dodecylsulfate-polyacrylamide gel electrophoresis. After proteolytic digestion, the radioactive oligosaccharides were sequentially liberated from the glycopeptides by treatment with different endo--N-acetylglucosaminidases and with peptide:N-glycosidase or, finally, by hydrazinolysis. In this manner, four groups of glycans could be obtained by consecutive gel filtrations and were subfractionated by HPLC. The structures of the individual oligosaccharides were analyzed by micromethylation, by acetolysis or by digestion with exoglycosidases. The major species amongst the high mannose glycans at Ans-406 of the viral glycopolypeptide were found to be Man1-2Man1-3(Man1-2Man1-6)Man1-6(Man1-2Man1-2Man1-3)Man1-4GlcNac1-4GlcNAc and Man1-3(Man1-2Man1-6)Man1-6(Man1-2Man1-2Man1-3)Man1-4GlcNAc1-4GlcNAc, while the complex glycans at Asn-478 are predominantly GlcNAc1-2Man1-3(GlcNAc1-2Man1-6)Man1-4GlcNAc1-4GlcNAc (lacking, in part, one of the outerN-acetylglucosamine residues) and GlcNAc1-2Man1-3(Gal1-4GlcNAc1-2Man1-6)Man1-4GlcNAc1-4GlcNAc.Abbreviation BSA bovine serum albumin - endo D (F,H) endo--N-acetyl-d-glucosaminidase D (F,H) - HA hemagglutinin (HA₁, large subunit of HA - HA₂ small subunit - FPV fowl plague virus - PNGase F peptide:N-glycosidase F - SDS sodium dodecylsulfate     

Characterization of the recombinant Clostridium pasteurianum ferredoxin and comparison of its properties with those of the native protein

Ferredoxins (Fds) constitute an important class of nonheme iron-sulfur proteins. One of the most studied Fds is the [8Fe-8S] Fd from Clostridium pasteurianum. The gene for this Fd has previously been cloned and sequenced. We report the expression of this Fd in Escherichia coli, and the characterization and comparison of this recombinant protein to the native Fd. We have found that the purified recombinant protein has the same enzymatic, redox, magnetic and electronic properties as the native Fd isolated from C. pasteurianum, which indicates that the two [4Fe-4S] clusters present in the Fd were correctly formed in E. coli.     

Atrial natriuretic peptide induces breakdown of phosphatidylinositol phosphates in cultured vascular smooth-muscle cells

Discrepancies exist between extent of guanylate cyclase activation by atrial natriuretic peptide (ANP) in cell-free systems and ANP-stimulated levels of cyclic GMP in whole cells, and also between receptor affinity and dose effectiveness of ANP. Therefore, we have investigated whether, in addition to receptor-coupled guanylate cyclase activation, other second-messenger cascade systems may be involved in mediating both an increase in cyclic GMP and the physiological response to ANP. Equilibrium 125I-ANP binding studies on cultured thoracic aorta smooth muscle cells revealed the existence of low-affinity (approximately 10(-8) M, 84.5 fmol/10(5) cells) and high-affinity (approximately 10(-10) M, 12.5 fmol/10(5) cells) binding sites. We confirm that ANP elevates intracellular cyclic GMP (EC50 approximately 10(-8) M) and inhibits agonist-(isoproterenol and forskolin)-induced increases in intracellular cyclic AMP (IC50 approximately 10(-9) M). ANP also stimulated breakdown of phosphatidylinositol phosphates and generation of inositol phosphates with a half-maximally effective concentration of approximately 10(-10) M. The extent of phosphatidylinositol polyphosphate hydrolysis was small (120%) in comparison to that of phosphatidylinositol (Ptd-Ins) (200%). Ptd-Ins hydrolysis was paralleled by the appearance of glycerophosphoinositol, and there was also a close temporal relationship between these processes and the accumulation of intracellular cyclic GMP. Smooth muscle cells released [3H]arachidonic acid label in response to ANP (EC50 approximately 10(-10) M). Taken together, the data suggest that the vasorelaxant hormone ANP has stimulatory effects on phosphoinositol lipid metabolism via both phospholipase C (generation of inositol phosphates) and phospholipase A2 (generation of releasable [3H]arachidonic acid and indirectly glycerophosphoinositol). In contrast, stimulation of phosphatidylinositol phosphate breakdown by the vasoconstrictive hormone angiotensin II is not associated with glycerophosphoinositol formation, and neither cyclic GMP nor cyclic AMP levels were influenced by this hormone.     

Contrasting evolutionary histories among tightly linked HLA loci

Genes comprising the major histocompatibility complex (MHC) play a central role in governing the immune response of vertebrates. A great deal of information has been revealed on the molecular biology and physiology of these loci, but three features-the high polymorphism, tight linkage among the loci, and the nonrandom association of alleles-make the system of particular interest from the perspective of population genetics. Information on the dynamic evolutionary forces that have acted on a locus can be inferred from the number and distribution of alleles that it carries. Ten loci from the HLA region of the human MHC, each sampled from several different populations, have been examined for departures from the expected value of homozygosity under the condition of selective neutrality. The homozygosities of five class I and II loci that code for membrane glycoproteins, HLA-A, -B, -C, -DR, and -DQ, and of glyoxylase I (GLO) were significantly less than the neutrality expectations. This suggests the presence of some form of balancing selection. In spite of being closely linked, in fact, located between the class I and class II histocompatibility loci, the homozygosities of the four class III or complement loci C2, Bf, C4A, and C4B, which are detected by electrophoresis, were indistinguishable from, or exceeded, that expected under neutrality. Although this conforms to the suggestion that, in general, electrophoretic variants are neutral, because of the tight linkage to loci demonstrating a history of selection, it is possible that the mechanism for generating variation in the class III loci may be different from that of the class I and class II loci.     

Anmerkungen zur Bestandsentwicklung des Uhus (Bubo bubo) in Bayern

Zusammenfassung In zwei Kerngebieten des Uhuvorkommens der Bundesrepublik Deutschland hat der Uhu in neuester Zeit scheinbar sehr stark zugenommen. In der Frankenalb (10 000 km²) schätzte man von 1955 bis 1964 etwa 30–40, Anfang der 70er Jahre etwa 50 und 1975–1980 75–83 Paare. In einem Ausschnitt der mittleren bayerischen Alpen (ca. 840 km²) waren um 1937 nur 3 Brutpaare bekannt, um 1975 mindestens 4; 1983/84 konnten jedoch mindestens 14 Brutpaare ermittelt werden. Die Zunahme ist zumindest größtenteils auf intensivere und systematische Kontrollen zurückzuführen. Der für die 30er Jahre vermutete Tiefstand des Bestandes in der Bundesrepublik dürfte weit höher gelegen haben als bisher vermutet, da zeitgenössische Quellen wahrscheinlich nur Teile des Bestandes erfaßten. Sicher lag der Bestand im 20. Jahrhundert wohl nie unter 50 Brutpaaren (bisherige Annahme 35), selbst zur Zeit des Minimums in der Bundesrepublik wahrscheinlich deutlich höher. Eine Obergrenze des bayerischen Brutbestandes von 200 Brutpaaren um 1980 ist durchaus realistisch. In den Alpen ist auch heute noch der Brutbestand völlig unzureichend bekannt. Das Aussetzen von Uhus hat in Bayern sicher keine Bedeutung für die Bestandsentwicklung gehabt. Der Median liegt für Todfunde ausgesetzter Uhus bei 0,43, für als Nestling beringte Wilduhus bei 1,9 Jahren. An Leitungen starben hochsignifikant mehr Wildvögel, an Straßen und Eisenbahnen dagegen hochsignifikant mehr ausgesetzte Vögel. Unterschiedliche Wanderneigung zwischen Wilduhus und ausgesetzten Vögeln ließ sich nicht sichern. Aussetzung und Wiedereinbürgerung sollten in Süddeutschland unbedingt unterbleiben und dürften auch im mittleren und nördlichen Deutschland mittlerweile überflüssig geworden sein. Eine Bestandsüberwachung und Kontrolle in ausgewählten Gebieten ist dagegen vordringlich.

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Some remarks on long term trends in the breeding population of the Eagle Owl (Bubo bubo) in Bavaria

Summary In two centres of its distribution in the Federal Republic of Germany the Eagle Owl has increased remarkably in recent years. Between 1955 and 1965 the population of the Frankonian Jura (ca. 10 000 km²) was estimated between 30 and 40 pairs, at the beginning of the seventies about 50, and in 1975/80 about 75–83 pairs. In an area of the Bavarian Alps (ca. 840 km²) only 3 pairs were known in 1937, at least 4 in 1975. In 1983/84, however, a thorough examination revealed at least 14 pairs in this area. The increase in both areas is most likely due to intensive and systematic checks in recent years. The lowest population level in the FRG, supposed for the thirties, was probably higher than estimated so far as contemporary sources apparently did not cover the whole population. During the 20^th century the population total of the FRG most likely has never been below 50 pairs and was probably higher even in times with minimum level. For 1980 an upper limit of 200 pairs in Bavaria seems to be realistic. In the Alps, however, the population size remains unsufficiently known even today. Release of captive bred birds didn't seem to have any increasing and stabilizing effect on the Bavarian population. The median for released birds found dead was 0,43, for wild chicks 1,9 years. Significantly more released birds died by traffic, whereas significantly more wild birds were victims of overhead lines. Differences in dispersal between wild and released birds seem likely, but could not be proved statistically. Releasing resp. reintroduction should be stopped in southern Germany; even in middle and northern Germany this conservation strategy seems to be unnecessary today. Survey and protection of breeding sites, however, should be continued or reorganized.