Amyloid fibrils derived from V-region together with C-region fragments from a lambda II-immunoglobulin light chain (HAR)

Amyloid fibril proteins were isolated from the spleen of a patient with IgD(lambda)-plasmocytoma by extraction and gel filtration in 5M guanidine hydrochloride. The molecular mass of the predominant polypeptide chain was approximately 5000 Da. Its complete amino-acid sequence was elucidated by stepwise automated degradation of the carboxymethylated polypeptide chain and by structural studies of tryptic and thermolysinolytic cleavage products. The length of the polypeptide chain was 58 to 59 residues and it was homologous to the amino acids in positions 8 through 65 of the variable part of an lambda-type immunoglobulin light chain, which was most closely related to the lambda II subgroup. The N-terminal sequence of this amyloid fibril protein proved to be heterogeneous, indicating cleavage after the amino acids in positions 7 and 8. Peptides from the constant part of the lambda-chain were unexpectedly found in the tryptic digest of the denatured amyloid protein HAR. One polypeptide derived from the constant region was separated from the main component by high performance liquid chromatography. Its amino-acid sequence commenced at position 111 and could be traced in 41 steps. In this case, at least two constant region fragments were shown to be constituents of the amyloid fibril protein. The association of fragments from the variable as well as the constant region is discussed with respect to amyloid formation.     

Identity of a cytosolic neutral cholesterol esterase in rat liver with the bile salt stimulated cholesterol esterase in pancreas

A neutral cholesterol esterase has been purified to homogeneity from the cytosolic fraction of rat liver. The 105,000 x g supernatant fraction of rat liver was applied to a DEAE-cellulose column to isolate a partially purified fraction of hepatic cholesterol esterase. Immunoblot analysis of the partially purified liver fraction with the anti-porcine pancreatic cholesterol esterase IgG demonstrated a single band with a molecular weight of 67,000. The hepatic protein was then isolated by immunoaffinity chromatography technique using a column constructed with antibodies prepared against the pancreatic cholesterol esterase. Characterization of the hepatic cholesterol esterase revealed that the hepatic enzyme shared antigenic epitopes with the pancreatic cholesterol esterase and was similarly activated by addition of bile salt such as taurocholate. Moreover, amino-terminal sequencing analysis of the hepatic cholesterol esterase showed an identical sequence with the pancreatic enzyme. Taken together, these results showed that the cholesterol esterases in the liver and the pancreas are very similar and possibly identical proteins.     

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     

Amyloid kidney stones of uremic patients consist of beta 2-microglobulin fragments

Urinary stones with amyloid structure, obtained from uremic patients, were analyzed according to molecular weight, amino acid sequence, and antigenic content. A major protein of approximately 7 kD, designated AB protein, was isolated by size exclusion using HPLC in 60% formic acid. AB protein reacted in immunodiffusion only with an antiserum to beta 2-microglobulin, with beta 2m spurring over AB protein. N-terminal amino acid sequence analysis defined two fragments homologous to beta 2m. One fragment commenced with Ile at position 7 and the other with Ser at position 20, with a cleavage point subsequent to a lysyl residue in both. It is concluded that beta 2m is a precursor of urinary amyloid stones and intratubular concretions of patients with preterminal and terminal renal failure; limited proteolysis is involved in AB amyloid generation.     

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.

     

Preferential expression of the maternally inherited X-linked phosphoglycerate kinase allele in human erythrocytes

Summary The stability of allelic gene expression of X-linked phosphoglycerate kinase was studied in seven carriers of a rare genetic variant named PGK München. The enzymatic activities in erythrocytes of five heterozygous females and three hemizygous males were determined repeatedly over a period of 10 years (1975–1984) and shown to remain constant. As the phosphoglycerate kinase activity is lower in cells expressing the PGK München allele, the ratio of the two cell types in all heterozygous females of the PGK München kindred could be calculated from the PGK activity and from the known allozyme activities in erythrocytes of homozygous wild type or hemizygous PGK München carriers. Since the maternal or paternal origin of both alleles is known from the pedigree, the quantitative expression of the maternally derived allozyme in heterozygous women could be determined. In heterozygous carriers the cell pool expressing the maternally inherited allele was significantly increased, independently, of the PGK allele linked to the maternal X chromosome (P<0.001). Our data show that inactivation of one of the two X chromosomes in human female erythropoietic stem cell precursors may be non-random, at least in the kindred and cell populations described here. The results are discussed in the context of random X chromosome inactivation (Lyon hypothesis).Dedicated to J.S., the senior of the family studied, on the occasion of her 80th birthday     

Spike synchronization of tympanic receptor fibres in a grasshopper (Chorthippus biguttulus L., Acrididae)

In recordings from single tympanic receptor fibres in C. biguttulus, the response to synthesized sounds (rectangularly modulated white noise) interrupted by very brief (a few milliseconds) gaps was examined. In behavioral tests, females of the species respond very differently to such 'model syllables' at moderate intensities, depending on the gap width. If the gaps (in a moderate-intensity syllable) are larger than 2 ms, the stimulus fails to elicit a response, whereas stimuli with gaps smaller than 1 ms are as effective as uninterrupted syllables (D. von Helversen 1972; O. von Helversen 1979). Neither the mean spike count nor the interspike-interval distribution of the single receptor response contains the information sufficient to distinguish uninterrupted syllables from syllables with gaps. On the other hand, examination of the temporal distribution of the spikes reveals that gaps (or the pulse onsets following the gaps) cause spike synchronization. An index of synchronization (IS) was defined as a measure of this gap-induced effect. Analysis of the receptor responses based on IS revealed differences that correspond quantitatively to the abrupt abolition of the behavioral response at a gap-width between 1 and 2 ms. From the hypothesis that such brief gaps are detected by the nervous system by way of spike synchronization in the tympanic nerve, one can predict certain features of the behavioral response to high-intensity stimuli. The gap-induced spike synchronization was more pronounced at higher temperatures. This effect was demonstrated in both summated recordings from the tympanic nerve and single fibre recordings. Experiments with primary auditory fibres of Locusta migratoria showed that the receptors in this species respond very similarly to the same stimuli. That is, the receptors of C. biguttulus are not specially adapted for detecting very brief gaps. Synchronization of the spikes in parallel receptor fibres of the tympanal nerve is probably a general feature of acridids; we infer that in C. biguttulus this gap-induced synchronized activity is detected by special processing in higher auditory centres.     

Uptake of U-14C-glucose by Streptococcus mutans in the presence of saccharin

The uptake of U-14C-glucose by resting cells of Streptococcus mutans OMZ-176 was studied in the presence of the artificial sweetener saccharin as well as sodium chloride. Glucose grown cells were resuspended in phosphate buffer (0.05 M, pH 7.8), and the uptake of U-14C-glucose was observed for 150 min in time intervals of 30 min, in the presence of 0.02 and 2.00 mg/ml of sodium saccharin as well as sodium chloride. As compared to the control and the sodium chloride treatments, sodium saccharin at the highest concentration range more than doubled the accumulation of radioactive labelled carbon within the cells.